Professional Documents
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Maria Duca
Plant
Physiology
Plant Physiology
BIOLOGICAL AND MEDICAL PHYSICS,
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Editorial Board:
Masuo Aizawa, Department of Bioengineering, Judith Herzfeld, Department of Chemistry,
Tokyo Institute of Technology, Yokohama, Japan Brandeis University, Waltham, Massachusetts, USA
Olaf S. Andersen, Department of Physiology, Mark S. Humayun, Doheny Eye Institute,
Biophysics and Molecular Medicine, Los Angeles, California, USA
Cornell University, New York, USA Pierre Joliot, Institute de Biologie
Robert H. Austin, Department of Physics, Physico-Chimique, Fondation Edmond
Princeton University, Princeton, New Jersey, USA de Rothschild, Paris, France
James Barber, Department of Biochemistry, Lajos Keszthelyi, Institute of Biophysics, Hungarian
Imperial College of Science, Technology Academy of Sciences, Szeged, Hungary
and Medicine, London, England Robert S. Knox, Department of Physics
Howard C. Berg, Department of Molecular and Astronomy, University of Rochester, Rochester,
and Cellular Biology, Harvard University, New York, USA
Cambridge, Massachusetts, USA Aaron Lewis, Department of Applied Physics,
Victor Bloomfield, Department of Biochemistry, Hebrew University, Jerusalem, Israel
University of Minnesota, St. Paul, Minnesota, USA Stuart M. Lindsay, Department of Physics
Robert Callender, Department of Biochemistry, and Astronomy, Arizona State University,
Albert Einstein College of Medicine, Tempe, Arizona, USA
Bronx, New York, USA David Mauzerall, Rockefeller University,
Britton Chance, University of Pennsylvania New York, New York, USA
Department of Biochemistry/Biophysics Eugenie V. Mielczarek, Department of Physics
Philadelphia, USA and Astronomy, George Mason University, Fairfax,
Steven Chu, Lawrence Berkeley National Virginia, USA
Laboratory, Berkeley, California, USA Markolf Niemz, Medical Faculty Mannheim,
Louis J. DeFelice, Department of Pharmacology, University of Heidelberg, Mannheim, Germany
Vanderbilt University, Nashville, Tennessee, USA V. Adrian Parsegian, Physical Science Laboratory,
Johann Deisenhofer, Howard Hughes Medical National Institutes of Health, Bethesda,
Institute, The University of Texas, Dallas, Maryland, USA
Texas, USA Linda S. Powers, University of Arizona,
George Feher, Department of Physics, Tucson, Arizona, USA
University of California, San Diego, La Jolla, Earl W. Prohofsky, Department of Physics,
California, USA Purdue University, West Lafayette, Indiana, USA
Hans Frauenfelder, Andrew Rubin, Department of Biophysics, Moscow
Los Alamos National Laboratory, State University, Moscow, Russia
Los Alamos, New Mexico, USA
Michael Seibert, National Renewable Energy
Ivar Giaever, Rensselaer Polytechnic Institute, Laboratory, Golden, Colorado, USA
Troy, NewYork, USA David Thomas, Department of Biochemistry,
Sol M. Gruner, Cornell University, University of Minnesota Medical School,
Ithaca, New York, USA Minneapolis, Minnesota, USA
Plant Physiology
123
Maria Duca
University of Academy of Sciences
of Moldova
Chişinău
Moldova
v
vi Preface
vii
viii Contents
4 Photosynthesis . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . ........ 65
4.1 Importance of Photosynthesis and the Global Role
of Green Plants. . . . . . . . . . . . . . . . . . . . . . . . . . ........ 68
4.2 The Leaf as a Specialized Photosynthesis Organ . . . ........ 70
4.3 The Structure, Chemical Composition, Function
and Origin of Chloroplasts . . . . . . . . . . . . . . . . . . . . . . . . . . 72
4.4 Photosynthesis Pigments . . . . . . . . . . . . . . . . . . . . . . . . . . . 77
4.5 Photosynthesis Energetics . . . . . . . . . . . . . . . . . . . . . . . . . . 81
4.6 Photosynthesis Mechanism. . . . . . . . . . . . . . . . . . . . . . . . . . 86
4.6.1 Light Phase of Photosynthesis . . . . . . . . . . . . . . . . . 87
4.6.2 The Dark Phase of Photosynthesis . . . . . . . . . . . . . . 101
4.7 Photorespiration . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 107
4.8 Endogenous Regulatory Elements of Photosynthesis . . . . . . . . 110
4.9 Ecology of Photosynthesis . . . . . . . . . . . . . . . . . . . . . . . . . . 117
References. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 122
Index . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 309
Chapter 1
Introduction to the Educational
Course of Plant Physiology
Historical Background
1727—St. Hales identifies the pathways of water, mineral salts and organic sub-
stances circulation.
1771—J. Priestley discovers photosynthesis.
1775—M. Malphigi describes the cycle of substances in plants—the ascending and
descending currents.
1800—J. Senebier edits “Plant Physiology” in 5 volumes.
1804—J. Senebier and Th. Saussure argue that photosynthesis represents the
nutrition of plants with carbon.
Brief Updates
During the last decades, by using gene engineering methods, plants with recom-
binant DNA have been created, also called genetically modified plants (GMPs), this
fact favoring the emergence of a new direction in plant physiology—the physiology
of transgenic plants which aims to determine the physiological and biochemical
changes of transgenic plants as a result of the inclusion of new genes into their
genome. Thus, the use of GMPs has allowed the elucidation of the genetic and
physiologic mechanisms of the activity of genes artificially included in the plant
1 Introduction to the Educational Course of Plant Physiology 3
genome, among which are also those that are normally found in animal organisms,
such as the Green Fluorescence Protein gene (GFP) from certain jellyfish species.
The GFP emits a green fluorescence under UV light, and its fusion with any other
protein allows the positional analysis of the last within the cell, the mechanism
being similar to that of radio-labeling.
Inserting auxine phytohormone biosynthesis genes (iaaM and iaaH) into the
tobacco genome resulted in more viable transgenic plants with a more active
vegetative morphogenesis and reproductive development and with both a higher
amount of water stored in tissues and a higher resistance to drought.
Another example is represented by the ferric superoxide dismutase gene
(FeSOD) from Arabidopsis thaliana (one of the genes involved in antioxidative
protection) which was included into the genomes of tobacco and wheat. The
genetically modified plants proved more resistant to the oxidative stress than the
control, confirming that the gene is expressed.
Lately, to study a particular gene function the antisense strategies are often
applied. The best known example is given by the gene that encodes the synthesis of
the polygalacturonase enzyme, involved in cell wall degradation in ripening tomato
fruits. After including this gene in the tomato genome, in reverse orientation, sense
and antisense RNA will bind on the basis of complementarity, thus obstructing
translation and leading to longer fruit preservation.
Plant physiology is a very important branch of biological sciences that studies the
life of plants—the laws and mechanisms of physiological and biochemical pro-
cesses, their significance, their interdependence with environmental factors in
ontogenetic dynamics. The notion of physiology originated from Greek by joining
the words physis, which means “function” and logos—“science”.
Plant physiology has appeared in 1800, when the Frenchman J. Senebier edited
his first monograph in five volumes “Plant Physiology”, which included not only
his own experimental results, but also those obtained in this scientific field by:
M. Malpighi, who has described the flow of substances in the plant (1775);
St. Hales, who demonstrated that water and mineral salts flow through the xylem,
while organic substances—through the phloem (1727); J. Pristley, who has dis-
covered photosynthesis (1771), etc.
During the development of plant physiology as a science, it has been based on
two directions: anatomical/morphological (descriptive) and physiological (experi-
mental), which, in principle, can be considered two basic research methods. This
division is relative, because vegetal organs can’t be studied without taking into
account their function, just as any processes cannot be studied without knowing the
4 1 Introduction to the Educational Course of Plant Physiology
structures they are localized in. Any physiological process should be regarded as a
product of long evolution, which forms the plant ability to adapt to variable
environmental conditions. The function has evolved in relationship with the
structure of the organism and the structure has stabilized under the action of
environmental factors and according to the function. Thus, to study respiration, it is
necessary to know the structure and ultrastructure of mitochondria, and to reveal the
mysteries of photosynthesis, a unique and specific process happening only in green
plants, it is important to know the structure and ultrastructure of the assimilatory
apparatus.
Most of the compartments of plant physiology have been delimited in the
nineteenth century and are valid even nowadays. These are:
1. Cytophysiology (plant cell physiology);
2. Water regime of plants (H. Dutrochet, H. de Friz, J. Sachs);
3. Photosynthesis (G. Busengo, M. Ţsvet, J. Pristley, K.A. Timireazev);
4. Mineral nutrition (I. Leibih, G.B. Busengo, D.N. Preanishnikov);
5. Respiration (A.S. Famiţsin, I.P. Borodin, L. Paster);
6. Growth and development (J. Sachs, A.S. Famiţsin);
7. Plant movements (T. Nait, J. Sachs, Ch. Darwin);
8. Irritation (B. Sanderson, Ch. Darwin);
9. Resistance to unfavorable factors (D.I. Ivanovski).
Thus, plant physiology as a distinct branch of biology, aims to study successively
all vital processes that occur in vegetal organisms. In the second half of the twentieth
century the basics of a new branch of plant physiology named self-regulation were
laid. The phenomena of self-regulation and coordination of physiological processes,
as well as other processes, are studied at all the levels of organization of living matter
(molecular, intracellular, at the levels of tissue, organ, organism, biocoenosis) the
mechanisms of implementation being diverse and specific.
Self-regulation (autoregulation) is the property of biological systems to maintain
the stability of the physical and chemical conditions of the internal environment, of
the structure and properties of the organism in their elementary form, all these in
conditions of a dynamic equilibrium. Autoregulation represents the process, which
minimizes various deviations in the biological systems (pH, viscosity, redox-
potential, etc.), resulting from the influence of causative agents. Therefore, the
capacity of the vegetal organism of carrying out vital functions amidst changing and
unfavorable environmental conditions is implemented.
Such a stability has a dynamic and active character. It is maintained by complex
mechanisms, which determine the coordinated physiological activity of different
organs, thus allowing autoregulation of plant growth and development, organism
temperature, raw sap composition, regeneration of damaged tissues, adaptation to
stress conditions, etc. (Figure 1.1).
Self-regulation ensures integrity and homeostasis of plant organisms, allows
harmonious growth and development and helps react adequately to the alternating
1.1 The Definition and Scope of Plant Physiology 5
Fig. 1.1 Scheme representing regulative and directive processes in living organisms (Polevoy
1989)
Dominant centers
Regulatory contours
Electrophysiological
Phyto hormonal regulation
regulation
Trophic regulation
Fig. 1.2 Interaction of regulatory systems (Polevoy 1989). Regulatory level: I intracellular, II
intercellular, III organismal
to the interaction between plant organs. Such an interaction can be observed during
cultivation of different vegetative explants in vitro. However, the existence of
trophic, hormonal and electrophysiological interactions between cells, tissues and
organs does not fully explain the behavior of a plant as a whole living organism.
There are higher level regulatory systems and mechanisms connecting organs and
functional systems of the plant during the life cycle of the organism and its onto-
genetic transitions.
The basic autoregulation mechanism at the organism level relies on the presence
of a few centers of dominance (the stem and the root apexes), which receive
information both from the external and internal medium and influence the living
organism by driving tissue morphogenesis, by creating physiological gradients,
polarity, channel connections (conductive fascicules), physiological oscillations.
1.1 The Definition and Scope of Plant Physiology 7
Glossary
Metabolism The totality of all the complex processes of synthesis (energy storage)
and degradation (energy release) undergone by the substances in a living
organism.
Morphogenesis Cyto-differentiation and development of visible structures (organs
or parts) in an organism during ontogenesis.
Levels of organization Systems with a specific organization (characteristic of
biological systems only) and with a character of universality.
Ontogenesis The series of transformations undergone by the organism, from egg
fecundation to death, according to the scenario for the respective species.
Respiration The process of oxidative degradation of complex organic substances
into inorganic ones accompanied by energy release.
References
Acatrinei Gh (1991) Reglarea proceselor ecofiziologice la plante. Editura Junimea, Iaşi, p 280
Burzo I, Toma S, Crăciun C ş. a (1994) Fiziologia plantelor de cultură, vol 1–4. Chişinău, Ştiinţa
Crăciun T, Crăciun L (1989) Dicţionar de biologie. Editura Albatros, Bucureşti, p 285
Derfling K (1985) Gormony rasteniy. Mir, 304 p
Duca M (1996) Sisteme şi mecanisme de autoreglare la plante. Chişinău, USM, 199 p
Duca Gh, Zănoagă C, Duca M, Gladchii V (2001) Procese redox în mediul ambiant. Chişinău, 381 p
Lebedev SI (1982) Fiziologiya rasteniy. M. Kolos, 544 p
Milică C, Dorobanţiu N. ş. a (1982) Fiziologia vegetală. Bucureşti, Ed. Didactică şi Pedagogică,
375 p
Polevoy VV (1989) Fiziologiya rasteniy. M. Vysshaya shkola, 464 p
Polevoy VV (1982) Fitogormony. L. Izd. Leningradskogo universiteta, 248 p
Tarhon T (1992) Fiziologia plantelor, vol I, II. Chişinău, Lumina
Udovenko GV, Sheveluha VS (1995) Fiziologicheskie osnovy selektsii rasteniy, vol 2. VIR
Yakushina NI (1980) Fiziologiya rasteniy. M. Prosveshchenie, 303 p
Chapter 2
Plant Cell Physiology
Abstract The cell is the smallest structural and functional unit of all living
organisms, at the level of which all the fundamental characteristics of life are
manifested. The cell is composed of several structures which have evolved to
perform unique functions:
• the cell wall contains a middle lamella, a primary cell wall and a secondary cell
wall (in order of their formation as a result of cell division) and is composed of
cellulose micro and macrofibrils immersed in an amorphous matrix consisting of
hemicellulose, pectic substances, proteins, but also of optional substances like
suberin and lignin that add up to its rigidity. The primary function of the cell
wall is that of a mechanical exoskeleton and delimiting barrier. The system of
interconnected gaps in the cell wall forms the apoplast which is a transport path
for liquids in the vegetal organism along with the symplast formed by the
plasmodesmata which connect the cytoplasm of the cells.
• the protoplasm is made of a viscous liquid matrix—the hyaloplasma which
serves as a medium for metabolic and energy exchange reactions, deposition of
substances, etc. and is also the place where the cellular organelles reside: the
nucleus, mitochondria, plastids, the endoplasmic reticulum, the Golgi body,
ribosomes, the vacuole.
• biological membranes (plasmalemma, tonoplast, membranes of the organelles)
represent fluid amphiphilic phospholipid bi-layers with various amounts of
embedded proteins which perform a diversity of functions and determine the
unique properties of the membranes. Biological membranes perform a variety of
functions like: compartmentalization, mechanical barrier, transport of various
substances including water (by osmosis), ATP synthesis (in chloroplasts,
mitochondria), receptor function, etc.
Cells constantly exchange substances with the external environment via active or
passive transport of the ions into or out of the cell. Active transport requires energy
and happens via vesicles, ion pumps and some carrier proteins, while passive
transport (no energy requirement)—via simple diffusion through the selectively
permeable membrane, or via facilitated diffusion through some carrier proteins or
protein channels. Water can also enter or exit the cell by means of osmosis,
Historical Background
1667—The cell was discovered by R. Hooke.
1838–1839—T. Schwann and M.J. Schleiden formulated the “Cell theory”.
1877—W. Pfeffer studied the osmosis phenomenon in vegetal cells.
1880—G.D. Thuret and J.B. Bornet discovered plasmodesmata.
1890—S. Altman discovered mitochondria.
1895—Ch. Owerton formulated the theory of protoplasm permeability.
1897—A. Garnier discovered the endoplasmic reticulum.
1898—C. Golgi discovered the dictyosomes.
1955—G.E. Palade discovered the ribosomes.
1958—R. Buvat launched the theory of vacuole emergence from the endoplasmic
reticulum.
1959—J. Robertson demonstrated the structural uniformity of all biological
membranes.
2 Plant Cell Physiology 15
Brief Updates
Cell death (apoptosis) represents the last stage in the life of a cell and is genetically
programmed, but can also be triggered by the action of various unfavorable factors.
The apoptosis process takes place when the metabolism ceases and is accom-
panied by some morphological and structural modifications—the destruction of the
genetic nuclear apparatus, of the mitochondria and chloroplasts, an increase in the
amount of Ca2+ ions, etc. The gravest changes occur at the DNA level, where
endonucleases cleave the macromolecule in small fragments of 50–300 kb. A
crucial role in the control of apoptosis is played by phytohormones (like ethylene)
and by DNA methylation. Serine and cysteine proteases are also involved in this
process.
There are very few data relative to how the apoptosis signal is passed from one
cell to another one. The H52-gene was discovered in tomatoes, which encodes an
HD-Zip transcription factor involved in this process. The artificial inhibition of this
gene’s expression leads to an imbalance in the control of apoptosis and to an
increase in the amount of ethylene and salicylic acid.
The induction of apoptosis in plants and animals share many common features.
One of them is the presence of the BI-1 gene homologues both in plants and
animals, for instance the Arabidopsis AtBI-1, which manifests itself under the action
of biotic (infections, pathogens) and abiotic factors (oxidative stress).
In some plant species (Arabidopsis thaliana, peas and rice) the dad1-gene was
detected which prevents apoptosis onset just like its analogue in the animal cell.
The activity of this gene decreases abruptly in senile plants and during seeds
formation. The inclusion of this gene in guinea pig cells delayed the apoptosis
onset. This fact demonstrates that the mechanism of apoptosis, especially its sup-
pression is similar in both plants and animals.
Living organisms that inhabit the planet consist of cells. The cell (from lat. cellula
or gr. cytos—“room”) is the smallest structural and functional unit of all living
organisms. Only viruses are recognized as non-cellular forms of life.
Cells vary in shape, size and color, but, whatever the cell type, it fulfills all the
criteria of living organisms, this phenomenon being demonstrated by cell cultiva-
tion on artificial media, their reproduction in vitro and even their possibility to
regenerate the entire plant.
16 2 Plant Cell Physiology
The cell was discovered in 1665 by an English physicist, Robert Hooke, who has
perfected the microscope and studied samples of cork. The cell theory has crys-
tallized during 1838–1839 owing to a number of German researchers: the botanist
M.J. Schleiden (1839) and the zoologist T. Schwann (1839), who recognized the
cell as the basic structural unit of living matter. Later R. Virchow (1855) confirmed
that cells could originate only from other cells—omnis cellula e cellula (Fig. 2.1).
The principles of cell theory are as follows:
The cell is a structural, morphological, functional unit of all living beings.
The cell is an open thermodynamic system, which constantly exchanges matter with
its environment and constantly transforms matter within the cell. All the eukaryotic
cells have a special complex of organelles that regulate metabolism, accumulate and
consume energy. The cell can exist as a separate organism (bacteria, protozoa,
certain algae and fungus species) or as a component of multicellular organism
tissues. Plants and animals consist of billions of cells, each of them specialized for
different functions (contraction, excretion, transport of substances and their depo-
sition, photosynthesis, etc.). More than 60 types of cells with various specialization
have been described in vegetal organisms.
Vegetal and animal cells have similar structure, functions and chemical
composition. Although the cell is the simplest form of life with a few microns in
diameter, its structure is very complex. In spite of this, the cell types share similar
functions and structure of cellular components.
The vegetal cell, unlike the animal one, has the ability to accumulate solar
energy, turning it into chemical or mechanical energy. Unlike other eukaryotic
cells, the vegetal cell is characterized by (Table 2.1):
• a system of plastids, present due to autotrophic nutrition;
• a central vacuole in the mature vegetal cell playing an essential role in osmosis
regulation and maintenance of the turgor pressure;
• a cell wall (cell envelope) which confers rigidity to the tissues.
2.1 The Cell as a Structural, Morphological, Functional Unit … 17
Table 2.1 Chemical composition of different plant cell organelles (% of dry matter)
Structural elements of the cell Proteins Lipids RNA
Cytoplasm 80–95 2–3 –
Plastids 30–45 20–40 0.5–3.5
Mitochondria 30–40 25–38 1–6
Ribosomes 50–57 3–4 35
Qualitatively, the chemical composition of the vegetal and animal cells is similar
(Fig. 2.2, Table 2.2).
Inorganic substances (water and inorganic ions) play the role of structural ele-
ments of various organic substances, as well as that of a reaction medium for
cellular metabolism. Traditionally, organic substances in vegetal cells can be
divided in the following groups:
• structural molecules, normally not involved in cellular metabolism and forming
instead the plant skeleton (cellulose, pectin, myosin, certain lipids, carbohy-
drates, proteins);
• enzymes—macromolecules catalyzing cellular metabolism (ribulose-1,5-
biphosphate carboxilase, phosphofructokinase, phospholipases, chlorophyllases,
amylases, ribonucleases, catalase, ascorbate oxidase, superoxide dismutase);
• micromolecular active substances (pigments, vitamins and phytohormones);
• micromolecular metabolites (resulting from specific catabolic reactions, serve
for the transport of electrons and protons, usually CoF);
• excretions deposited in the cell wall and vacuole (tannin, anthocyanin, lignin);
• reserve substances (starch).
18 2 Plant Cell Physiology
Table 2.2 Comparing the structure and composition of plant and animal cells (Pickering 1998)
Characteristics of an animal Common features of animal Characteristics of a vegetal
cell (the result of and vegetal cells (with regard cell (the result of autotrophic
heterotrophic nutrition) to the processes of life nutrition)
maintenance)
In animal cells secretory The cell membrane The cellulosic cell wall offers
vesicles containing cellular surrounding the cytoplasm support and protection against
products like hormones or controls the entrance and exit potential lesions caused by
enzymes can be often found of soluble substances being, water flowing into cells due to
thus, responsible for cell the osmotic force
content separation from the
environment
The cytoplasm of animal cells The cytoplasm contains Chloroplasts contain a special
is more dense and contains water, soluble substances like pigment called chlorophyll
more cellular organelles amino acids and (which absorbs light) and the
carbohydrates and supports enzymes necessary for
cellular organelles. Various glucose production via
metabolic reactions occur in photosynthesis
both the cytoplasm and
cellular organelles
Vacuoles are small and The nucleus contains the A big, permanent vacuole
temporary. They may be genetic material (which contains the water needed to
involved in digestion (e.g. makes up the genes and generate the turgor pressure
phagocytosis) or excretion chromosomes and encodes and is also a repository for
processes (contractile genetic information). ions and molecules
vacuoles can remove water Chromosomes become visible
excess from the cell) only during cell division
Carbohydrates are stored in Carbohydrates are stored in
the form of glycogen the form of starch (found in
the cytoplasm and in
chloroplasts)
Each daughter cell originates from the mother cell after division. The cel-
lular theory is considered to be one of the three greatest discoveries of the 19th
century, along with the law of mass and energy conservation and transformation
(A.L. Lavoisier, M.N. Lomonosov) and the theory of evolution (Ch. Darwin).
The cell wall (cellular envelope) is a complex formation with 3 basic layers:
• the middle lamella;
• the primary cell wall;
• the secondary cell wall.
2.2 Structural Organization, Chemical Composition ... 19
The middle lamella appears during the telophase of cell division (mitosis)
between two daughter cells as a product of the Golgi apparatus activity. For young
growing cells from meristematic tissues a primary cell wall is characteristic, and
while aging, the secondary structure of the cell wall emerges, both being produced
by the protoplast (Fig. 2.3).
Functions of the cell wall:
• represents a mechanical exoskeleton for the cell;
• confers shape to the cell and rigidity to vegetal tissues;
• prevents the rupture of the cytoplasmic membrane due to hydrostatic forces
acting from the cell interior;
• represents a barrier for various infections;
• participates in the absorption and transport of water and mineral salts;
• represents a specific ion exchanger;
• participates in the exchange of substances;
• lectins contained in the cell wall recognize symbiotic bacteria, which cause the
formation of root nodules in plants.
Properties. The cell wall has a high rigidity but can also support elastic
deformations. The thickness of the cell wall in different plant species are ranging
from 0.1 to 10 µm.
Chemical composition. The main components of the cell wall are: cellulose,
hemicellulose, pectic substances, proteins, etc. Cellulose molecules (C6H10O5)n
represent long unbranched chains, consisting of 3–14 thousand glucose residues.
20 2 Plant Cell Physiology
The cellular protoplasm, or cytoplasm (from gr. cytos “cavity” and plasma
“structure, substance”) represents the fundamental mass of the cell. Optical
microscopy reveals cytoplasm as a viscous, transparent, colorless, homogeneous
liquid, immiscible with water, characterized by surface tension, constant pH and rH
values, semipermeability and selectivity, excitability, viscosity and movement, in
which a series of organelles with varying size, shape, structure, chemical compo-
sition and functions.
Electron microscopy, reveals cytoplasm as a heterogeneous and complex
structure. It consists of a cytoplasmic matrix—the hyaloplasma, in which the cel-
lular organelles are suspended (Figs. 2.4 and 2.5).
Hyaloplasm (from gr. hyalos “transparent, glass”) represents the soluble phase,
which performs a series of functions:
• a matrix for metabolic and energy exchange reactions (glycolysis);
• deposition of organic (glycogen, starch) and inorganic substances;
• cell movement;
• cell adaptation to environmental conditions.
The hyaloplasm includes:
• a structural part, representing several types of fibrillar and globular proteins,
arranged in microfilaments (with the diameter of 6–10 nm) or in microtubules
Fig. 2.4 The components of the vegetal cell (Yakushina N.I., 1980)
22 2 Plant Cell Physiology
Fe2+, Mn2+, Cu2+, Zn2+, NH4+, CI−, PO43−, SO42−) or ions bound by organic
molecules and, finally, inclusions (reserve substances).
The ratio between these electrolyte and non-electrolyte substances contribute to
the formation of a particular physical-chemical state of the hyaloplasm, leading to
autoregulation of metabolic processes in the cell and maintaining the physiological
and biochemical homeostasis, characterized by perfect integrity.
As the hyaloplasm contains filamentous protein macromolecules it is more fluid
when globular proteins predominate and more viscous if fibrillar proteins are present
in big quantities, in the last case a transition from a sol to a gel state being possible.
The intensity of metabolic processes is inversely proportional to viscosity and is
often indicative of adaptation phenomena (to cold, drought, humidity). Protoplasm
viscosity. is 16–20 times higher than water viscosity. It also changes during onto-
genesis The lowest values of viscosity were detected during flowering while the
highest ones—in seeds during maturation and anabiosis. The hyaloplasm is in
constant motion, which can be circular or sliding. It is characterized by a specific
isoelectric point, because the major part of its content is represented by proteins.
Several cellular organelles are incorporated in the hyaloplasm. They have dif-
ferent shapes and sizes, perform specific functions and provide the vital activity of
the vegetal organism. These organelles can be divided into two major groups:
(a) membranous organelles (bi-membranous and uni-membranous);
(b) non-membranous organelles, which have no phospholipid membrane on their
surface.
The membranes of the cellular organelles have a thickness of 6–7 nm and can
be classified into:
• double (internal and external);
• simple.
Double membranes are characteristic for the nucleus, mitochondria and chlo-
roplasts, while the simple membranes can be found in the endoplasmic reticulum,
the Golgi apparatus, in lysosomes and vacuoles.
Properties of biological membranes. Biological membranes represent a
homogeneous and transparent film that forms a system of folds, which increases the
surface area. The thickness of the membranes varies from one organelle to another
(from 5.5 to 20 nm). The fine structure of biological membranes reveals the
property of selective permeability, this fact contributing to a strict control of matter
inflow and outflow. For example, the plasmalemma is slightly permeable for glu-
cose, while the tonoplast is impermeable. Thus, the products of photosynthesis are
transported from the cell and do not accumulate in the vacuole.
After cell death, biological membranes lose their property of semipermeability
which proves that their structure and chemical composition are maintained by
energy consumption.
Chemical composition. Biological membranes are composed of lipids (1/2),
proteins (1/3), polysaccharides (in small amounts), enzymes and various ions. The
protein/lipid ratio of a membrane reflects the intensity of its functional activity.
The most abundant lipids are phospholipids, followed by glycolipids and cho-
lesterol. Sterols and unsaturated fatty acids confer porosity to the phospholipid
double-layer. Phospholipids are amphiphilic which means that the molecules have
one hydrophilic (polar) end and another highly hydrophobic end. This property
allows lipids to self assemble in a two-layered structure in the presence of water.
Lipids are not fixed stably in the membranes. They change permanently their
location by lateral diffusion (within the monolayer) or by transverse diffusion (“flip-
flop”) occurring between two distinct lipid monolayers (this occurs much less
frequently). Plasmalemma contains 35–40 % of lipids, mitochondrial membranes—
up to 28 %, while the myelin membranes—up to 80 %.
The proteins of the biomembranes (enzymes, receptors, pumps, channels, reg-
ulatory and structural proteins) consist mainly of hydrophilic (polar) amino acids,
which are responsible for some specific properties. Membrane proteins are char-
acterized by the ability to move within the fluid double lipid layer and can be
divided into 3 groups:
• Integral—which form hydrophobic links with the lipids and pierce the double
lipid layer;
• semi-integral;
• peripheric—proteins are on the surface of the biological membranes.
Biological membranes may also contain heterogeneous macromolecules (gly-
coproteins, glycolipids) and several minor components (coenzymes, nucleic acids,
antioxidants, carotenoids, pigments, etc.) depending on the role they perform.
2.4 Structure and Function of Biological Membranes 25
Fig. 2.6 Ultrastructure of the cellular membrane (Johnson and Raven 2002)
Fig. 2.7 Functions of the components of biological membranes (Johnson and Raven 2002)
The vital activity of each living organism is determined by its provision with
nutrients and water. A constant exchange of substances between the exterior and
interior environments occurs on the cellular level during its entire life cycle. Flow of
substances inside the cell is called absorption. The transport of substances from the
cytoplasm to the vacuole or to the lumen of xylem vessels is called secretion. If ions
or other substances are leaving the cell, this process is called excretion or desorption.
membrane (which prevents charged ions from passing through with varying effi-
ciency depending on ion charge, diameter).
With the advent of phospholipid membranes, transmembrane ion transfer
mechanisms have emerged (Fig. 2.9). When lipids predominate in the membranes,
permeability is higher for organic substances, while, if they are composed mainly of
proteins, permeability is higher for water and mineral ions. Ion transport across
biological membranes, may be passive or active.
Passive transport of ions occurs according to the chemical and electrochemical
gradients, without metabolic energy consumption. This phenomenon is based on
simple and facilitated diffusion.
Simple diffusion is the movement of ions and molecules along the concentration
gradient which is from a region of higher concentration to one of lower based on the
kinetic energy of the molecules, which increases with temperature and concentra-
tion. From the thermodynamic point of view, the diffusion vector is determined by
the chemical potential of the substance. The higher the concentration of the sub-
stance, the higher its chemical potential.
Passive transport across phospholipid membranes can occur with substances
which are soluble in lipids or if the molecular diameter of the molecules is smaller
than the pore diameter. The transport rate is inversely proportional to the diameter
and mass of the transported molecules (the ultrafilter concept elaborated by W.
Ruhland) and directly proportional to their solubility in lipids (the liposolubility
concept developed by Overton). An example of passive transport is the movement
of carbon dioxide from the air into leaf tissues.
2.5 Exchange of Substances Between the Cell and the Medium 29
The transport of polar molecules and ions along the concentration gradient takes
place with the help of transport proteins via facilitated diffusion. These proteins can
be integral to the membrane or they can reversibly bind the transported molecules,
cross the biological membranes and release the cargo, for instance, within the cell,
and the protein travels back to reinitiate the cycle.
Active transport is performed contrary to the concentration gradient with
metabolic energy consumption (ATP, NADH, NADPH) derived from the respira-
tion process.
Active transport can occur via specific transport proteins included in the bio-
logical membranes and forming protein carriers or ion pumps. Transporters in this
case have functions similar to those of enzymes. Absorption on one side of the
membrane and desorption on the other side, are very selective due to the high
specific affinity of the transporters for the cargo (e.g. ions). This transport type is
characteristic for carbohydrates, amino acids, nucleotides, etc. and may be simple,
in the case of one substance being translocated in a particular direction (Fig. 2.11),
or coupled (cotransport)—when the simultaneous transport of two substances takes
place (Figs. 2.10 and 2.12). When two molecules are transferred in the same
Fig. 2.10 Sodium and potassium ion transport carried out by a single pump (the K+/Na+-ATP-ase)
(Johnson and Raven 2002)
30 2 Plant Cell Physiology
Fig. 2.11 Active transport of a single substance by the H+-pump through a series of
conformational modifications
Fig. 2.12 Transport of two substances in different directions (Na+ and K+) and in the same
direction (Na+ and carbohydrates) (Johnson and Raven 2002)
Water exchange between cells and the environment is achieved through the inter-
vention of the following physical phenomena, characteristic of both the biological
systems and the non-living matter:
• water currents
• diffusion
• osmosis
• electroosmosis
• imbibition
• suction force of the cell.
Water currents represent the total movement of water, based on its free energy
(the water potential, which alternates depending on several physical factors). Water
moves from the region with higher potential of water to the region with a lower one.
Diffusion is the random movement of molecules from a region of higher con-
centration to one of lower concentration until the balance is achieved. The rate of
diffusion is determined by the temperature, the number and size of the particles, the
fluidity of the medium, etc. This is a slow process that occurs at small distances in
solutions with high difference in the potential of a chemical species (Fig. 2.13).
Osmosis (from lat. osmos—“impulse, urge”) represents the movement of water
or solvent molecules through a selectively permeable membrane. The direction of
this movement is from a lower concentration to a higher one, i.e. from a higher
water potential to a lower one (Fig. 2.14).
The phenomenon of osmosis is the fundamental way of water flow into the cell,
being very important for directing and distribution water in living organisms. The
resistance force that opposes water entrance into the cell is directly proportional to
the water concentration of the cellular juice and is called osmotic pressure (P).
Osmosis is responsible for water flow from the tissular fluids into the cells, from the
the soil (the accessible water) to the tiny root hairs, from the xylem into the cells
that form the leaf mesophyll. The lowest osmotic pressure (0.1 atm) can be
observed in aquatic plants and the highest pressures—in halophytes containing a
high percentage of mineral salts in the cellular juice. In most crop plants the osmotic
2.5 Exchange of Substances Between the Cell and the Medium 33
Fig. 2.15 Types of solution depending on the osmotic pressure. a Isotonic solution b Hypertonic
solution c Hypotonic solution
pressure values range from 0.5 to 3.0. Usually, the osmotic pressure of terrestrial
plants is in the 506–1010 kPa limit, while for aquatic plants—in the 101–304 kPa
limit. A high osmotic pressure—2,026–4,052 kPa—is identified in fruits, vegeta-
bles, berries, sugar beet.
The osmotic character of water exchange between the cell and its environment is
manifested in the phenomena of turgidity and plasmolysis, which depend directly
on the tonicity of the internal environment of the cell. Depending on the chemical
potential value, different situations may occur in the process of water exchange
between cells and the environment, resulting in different types of solutions—iso-
tonic, hypertonic and hypotonic (Fig. 2.15).
In isotonic solutions ion concentration inside the cell is equal to their concen-
tration outside the cell, i.e. the cells have the same osmotic pressure as the envi-
ronment. In such solutions there is no active water movement except for diffusion
and occasional short-distance water travel.
In hypertonic solutions ion concentration in the cell is lower than in the external
environment. Thus, water is eliminated from the cell by exosmosis causing
detachment of the plasmalemma from the cell wall (a phenomenon called plas-
molysis) and, consequently, plant wilting.
34 2 Plant Cell Physiology
The plasmolysis can be concave (early) and convex (final). In convex plas-
molysis the protoplasm is contracted around the vacuole and its contact with the cell
wall is maintained only via fine threads (Hecht’s filaments). Plasmolysis forms and
types are determined by the degree of protoplasm hydration and by the micro-
structural features of the plasma membrane (Fig. 2.16).
Hypotonic solutions are characterized by a higher concentration of ions in the
cell as compared to the external environment. This phenomenon contributes to
endosmosis, i.e. water molecule active movement from outside into the cell,
causing deplasmolysis, leading to an increase in cell turgidity. If the plant cell is in a
hypotonic medium, water enters it through the endosmotic flow and the cell
expands. As this happens, the cell wall exerts an increasing counter-pressure on the
cellular content. This is the turgor force (T), that gives the cell the state of rigidity
and tension. T represents the hydrostatic force of the intracellular solution exerted
on the plasmalemma, which shapes and stiffens the vegetal cell. The turgor force
reaches its maximum value in the morning and its minimum—in the afternoon
ranging from 5–10 atm in thallophytes to 100 atm in fungi. If a drought or intensive
transpiration takes place, the turgor force value reduces to 0 and the plant wilts.
Plasmolysis and deplasmolysis are phenomena characteristic only of living cells,
which have maintained semipermeability. The speed and form of plasmolysis
characterize the viscosity of the protoplasm. Plasmolysis allows to determine
osmotic pressure values, this fact being important in ecological research and
explains the ability of plants to absorb water from soil and retain it. Osmotic
processes in plants are of primary importance in ensuring water delivery through
absorption, efficient water circulation and the exchange of substances.
Electroosmosis is the movement of liquid through the pores of a membrane
under the influence of an electric field. This movement is possible because an
electric potential can install in the vegetal cell at the cell wall level due to pectin and
other substances containing carboxyl groups that dissociate in COO− and H+.
COO− ions give a negative charge to the cell wall. In order to reach an electrostatic
balance, ions of opposite charge are attracted and arranged parallel to the mem-
brane, thus, an electrical gradient being established. Biological membranes are
characterized by a certain electrochemical potential that contributes to water
circulation.
Imbibition forces are those which contribute to water penetration through
macromolecules of protein colloids and cellulose-pectin microfibrils. Imbibition is
based on colloidal and capillary effects and causes an irreversible increase in vol-
ume and mass. Imbibition based on colloidal effects predominate in the protoplasm,
but both types are characteristic for the cell wall. This process is very important
during seed germination, as the organic reserve matter, being hydrophilic, has the
ability to bind a large number of water molecules, which helps develop in seeds an
inner force of up to 1,000 kPa. Cellular proteins have a higher imbibition capacity
while cellulose has a weaker one. Each OH− radical of the β-glucose residues that
make up cellulose, can fix 3 water molecules while the acidic COOH group and the
NH2 amino group can retain 4 water molecules. Imbibition may be limited, when
2.5 Exchange of Substances Between the Cell and the Medium 35
Fig. 2.17 K. Hoffler’s diagram representing the value of cellular suction force as a function of cell
volume (starting with a state of plasmolysis and ending with a state of maximum turgor) (Milica
1982)
the imbibed mass remains in the gel state and unlimited when the colloid transitions
entirely into the sol state.
The totality of forces that contribute to water absorption in the cell form the
suction force, marked with S. It is determined by the difference between the osmotic
and turgor pressures. This force depends on internal and external factors, on the plant
species as well as on environmental conditions. As the turgor tends to maximum
values, the rate of water entrance into the cell due to the osmotic force reduces
gradually (S = P−T), until an equilibrium state is reached (P = T). In this case the
turgor pressure is equal to the osmotic pressure and the suction force value is 0.
During endosmoses the cell manifests itself as a self-regulating osmotic system,
because the relationship between the turgor force and the osmotic pressure deter-
mines the absorption intensity and the volume of water that flows in it. This
relationship was portrayed in a diagram by the German scientist K. Hoffler
(Fig. 2.17).
Glossary
Apoplast The totality of interfibrillar gaps, the so-called free space of the cellular
envelope. Water and various substances circulate through the apoplast from cell
to cell throughout the plant body.
Cellulose (C6H10O5)n A polysaccharide that represents the basic component of the
plant cell wall, and is usually in the form of fibrils. Its molecules are grouped into
micelles, microfibrils and macrofibrils, forming the housing for the membrane
36 2 Plant Cell Physiology
References
depends on their ionic balance, on the humidity of the environment, CO2 con-
centration, exposure to sunlight, phytohormone action. According to their water
regime plants have been classified in hydatophytes, homeohydrophytes and
poichilohydrophytes.
Historical Background
1634—JB. Van Helmont concluded that water is a constituent of the organic mass
of the cell.
1664—J. von Sachs demonstrated the role of temperature in water absorption by the
root.
1671—M. Malpighi discovered stomatal cells and their function.
1727—S. Hales measured for the first time the intensity of plant transpiration.
1837—R.J.H. Dutrochet discovered the osmosis phenomenon.
1901—H.H. Dixon worked out the theory of the cohesion force.
1926—N.A. Maximov formulated his theory of plant resistance to drought.
1976—L.N. Babuşkin discovered the absorption mechanism of water vapor from
the intercellular spaces in plants.
3 Water Regime 41
Brief Updates
The most important data that allow to understand the mechanism of water move-
ment in living organisms were revealed with the discovery of the membrane pro-
teins called aquaporins. Aquaporins belong to a large family of proteins
homologous to the major intrinsic bovine protein (MIP) and are characterized by a
structure containing integral membranous domains and by two repeats of a highly
conserved amino acid sequences Asn-Pro-Ala. These proteins were discovered in
plants in the late 80s of the twentieth century, concluding that some of them can act
as water channels and can facilitate the diffusion of an enormous water amount
along the transmembranous gradients.
The research that had followed the first discovery of aquaporins found that they
are very different in plants. Thus, the aquaporins of Arabidopsis are encoded by
more than 30 homologous genes. Based on the homology between sequences,
aquaporins biosynthesis genes can be classified into three subfamilies specific to
plants, two of which correspond to a specific subcellular localization (for instance
intrinsic to the plasma membrane or to the tonoplast).
It is quite surprising that some aquaporins are multifunctional proteins and,
besides water transport, may be also involved in the transport of osmosis-com-
patible solutions, like glycerol. Involvement of aquaporins in the transportation of
gases like CO2 or NH3 is also possible, this process taking place in leaves or in
symbiotic root nodules.
Water is the main mineral compound of the living matter, on which practically all
the vital processes depend and which maintains the normal physical state of the cell.
The physiological function of the plant organs are only possible when the cells are
saturated with water. Water is a most important constituent of plant life due to the
physical and chemical properties it possesses. In the process of homeostasis
maintenance and cell composition formation, water (Fig. 3.1) fulfills multiple roles:
• a basic solvent for mineral salts and organic compounds and, at the same time, a
dispersion medium for colloidal macromolecules and a medium for biochemical
reaction progression;
• an important factor in maintaining the stability of plant temperature helping to
avoid tissue overheating, which could arise due to the heat released during the
metabolic processes or under direct sunlight (summer);
• an element of protoplasm structure, which is fixed electrostatically among the
long catenae of polypeptides, allowing the physical and chemical properties of
the protoplasm, favoring the formation of colloidal systems and determining the
conformational structure of the proteins crucial for their functioning and also
necessary to ensure the maintenance of the ultrastructure and the functional
42 3 Water Regime
Total water content in plants is highly variable and depends on plant species and,
within the same species,—it depends on the organ, tissue, ontogenetic phase, etc.
Thus, algae contain 94–98 % of water, succulent leaves—95 %, reserve organs—
85 %, leaves—80 %, dry seeds—12–14 %. Environmental factors and the organ
type may influence the hereditarily expected values for this index. The variability of
water content in this case is determined by the water retention capacity of the plant.
Water retention is caused by osmotic forces, colloidal and capillary imbibition
forces, etc. Protoplasmic colloids have a higher capacity to retain water in young
leaves compared to older ones.
Water in plant cells is retained in the cell wall, the protoplasm (up to 90–95 % of
water), the vacuole sap (98 %). The amount of water retained by cellular envelopes
depends on its thickness, structure and chemical composition. Approximately
7–8 % water is bound to cellulose polymeric chains and is retained by superficial
bonds. The vacuolar sap contains up to 98 % water, which is retained by osmotic,
electroosmotic and imbibition forces.
Water is also a structural component of biological membranes—water interact-
ing with the membrane surface, water located in the space between the internal and
external chondriosome (mitochondria) membranes.
There are 3 water aggregation states that can be found in plants. Water in liquid
state is the basic component of all cells, because it is a component of the mem-
branes (30–35 % of the membrane weight), of the protoplasm and vacuole. In its
gaseous state (vapors), water can be found in intercellular spaces and in all the
aeriferous tissues. Water in the solid state of aggregation represents ice crystals,
formed during severe frost in intracellular and especially in intercellular spaces.
Intracellular crystals break cytoplasmic membranes deteriorating the cells.
Liquid water in the vegetal organism can be free (95 %), representing the basic
solvent for mineral and organic substances, ensuring colloid micelle dispersion in
the cytoplasm, or bound (4–5 %), retained by hydrogen bonds or by other types of
chemical bonds or immobilized in fibrillar structures of macromolecules (Fig. 3.2).
Free water is retained weakly in the plant organism. It circulates very easily in
vacuoles, cytoplasm and conducting vessels either inside the cell, or from cell to
cell, enabling, at the same time, turgidity. Free water represents the medium where
the biochemical processes take place and it often directly participates in these
reactions. Free water freezes at temperatures down to minus 10 °C, so plants with
high content of free water are less resistant to low temperatures.
Bound water is retained in plants very strongly. This water type is made up of
immobile molecules with no possibility for diffusion or evaporation, it is hardly
released by the cell. Bound water freezes at temperatures lower than −10 °C. It does
not circulate in the cell or in the entire plant, doesn’t take part in biochemical
processes and in dissolving organic or inorganic substances. Due to inability to act
as solvent, bound water doesn’t participate in the transfer and circulation of
substances.
44 3 Water Regime
There are numerous capillary spaces in plant cells (in vacuoles, in the spaces
between the colloidal micelles of the membrane and the protoplasm (while at the
tissue level—especially in the conducting vessels) in which the capillary forces
retain water molecules. This fraction is called capillary water.
There is also the constitutional water, chemically bound by certain molecules.
Release of this water by molecules implies their destruction.
The notions of free and bound water are relative, because these two forms of
water can transition one into another. In unfavorable environmental conditions,
when the vital activity of the plants is essentially reduced, the amount of free water
decreases, while the amount of bound water increases, resulting in a higher stress
resistance in plants. Quantitatively, free water always prevails when compared to
bound water, however in drought conditions this difference is smaller. The critical
limit of cell dehydration is 35 %, when vital processes are reduced to a minimum.
According to its origin, water can be exogenous and endogenous. Most of it is
of exogenous origin, absorbed by plants from the soil through the root system or, in
a small amount, from the atmosphere in the form of vapors. Plants can obtain water
through its aerial organs—leaves (from dew, rain water). During the drought period
the dew has a marked impact on young leave hydration—50–70 % H2O, but less so
in the case of old leaves—5–7 % H2O.
Endogenous water is synthesized during the process of respiration in
mitochondria.
The amount of water absorbed by plants depends not only on the root system size,
but also on the amount of water available in the soil and on the forces with which
the last retains water. Water retention forces depend on the osmotic pressure of the
solution present in the soil. Water can be found in many forms in the soil:
• Constitutional water (crystallization water) enters in the composition of organic
or inorganic molecules from the soil as crystallization water: for instance
CuSO4·5H2O, Na2CO3·10H2O etc. This water form can’t be used by plants due
to its huge retention force.
• Hygroscopic water forms a very thin hydration layer on the surface of soil
particles and is retained by very high forces (approximately 10,000–31 atm.).
This water form can’t be used by plants, its removal from the particle’s surface
being possible only by drying at 105 °C.
• Pellicular water from the surface of soil particles is retained by the hygroscopic
water film with forces higher than 30 atm, while the external layers of the film—
with 0.5–30 atm. Plants can absorb only a certain part of pellicular water (that
from the peripheral layers). Exceptions are some halophyte species that absorb
water from the deeper layers as well.
46 3 Water Regime
During the evolutionary process, vegetal organism saturation with water has been
possible permanently and sufficiently only for submerged plants (algae). Terrestrial
plants, by contrast, are living in an aerial environment and loose a substantial
amount of water through transpiration.
Angiosperm plants have attained a continuous flow of water in their organisms,
in order to keep the protoplasm of the assimilatory and other tissues saturated and to
counter water loss through transpiration. This has been possible due to the devel-
opment of a strong root system, of a dense network of conducting vessels, capable
to deliver water in the most remote parts of the plant and due to the development of
specialized protective tissues that protect the aerial parts against water loss.
Plants absorb water from the substrate continuously during the entire life cycle
through the root system, but can retrieve it also from precipitations, from fog or
from dew with its aerial organs. Water is removed in a proportion of 99 % through
transpiration and guttation processes. Continuous water currents between the root
system and aerial organs represents an essential requirement for the metabolic
activity and therefore for plant survival. In this case the root system plays the most
important role in plant water supply.
3.4 The Root System as a Specialized Organ for Water Absorption 47
Root system functions: fixation in the substrate (laminaria rhysomes fulfill only
the function of mechanical fixation), delivery of water and minerals, respiration,
assimilation, substance storage, tissue regeneration. For an efficient execution of
these functions, plants have developed the ability to orient in space, to respond to
gradients of vital factors, to form a maximum area of contact with the soil.
Morphological features: root system growth and development into the depth of
the soil and close to the surface.
The root system of plants varies in size and shape. The total length of the roots
with their branches reaches some dozens of meters even in small plants. The total
surface of the root is hundreds of times bigger than the surface of the stem and the
leaves. The contact surface of the root system with soil particles, as well as the
depth of root penetration into the soil vary in different species (alfalfa (lucerne)—
0.3 m, grapevine—18 m, spring wheat—2 m, potato—0.5 m) (Fig. 3.3).
Those plants that have adapted to intense sunlight (heliophytes) lose more water
through their aerial organs and have a much more developed root system than
plants adapted for diffused light (sciophyts), which always grow in wet places and
have a weakly developed root system.
V.G. Rotmistrov, V.A. Kolesnik, A.L. Modestov had investigated the actual size
of the root system. The length of all roots in cereals is 10,000 m, with 14 billion
absorbing hairs. An apple tree, which has 10 branches on the surface, develops
45,000 branches at the root system level.
The root system is developing better in a structured soil with sufficient humidity
and adequate aeration. Root system formation can be stimulated through a set of
actions like soil irrigation and fertilization and a series of agricultural techniques.
48 3 Water Regime
Anatomical features: the absorption zone, Kaspari stripes and passage cells
(Fig. 3.4).
Water absorption in superior terrestrial plants is carried out by roots, and namely
by the fine and always young root endings—the root hairs. Root hairs have a length
of 0.15–8.0 mm and a thickness of 0.1 mm. They are formed at a short distance
from the root tip in the area, where root growth in length ceases and they originate
from the elongated external cells of the rhyzoderm. To the internal wall of the root
hairs the protoplasm and the nucleus are localized, the interior is occupied by a
large vacuole and a vacuolar sap that is more concentrated than the electrolyte
solution in the external environment; thus a relatively high water permeability is
conditioned. Root hairs are regenerating every 2–3 days, determining the contin-
uous activity of the absorbing area. In Pinus silvestris up to 220 hairs per 1 mm2 can
be found, in Secala cereale—about 2,500. The apical meristem can produce 200–
400 new cells per day (Vicia faba) or up to 2,100 (Zea mays), in conditions of
sufficient water and nutrients.
The membranes of root hairs lack cellulose, but contain callose, which is a
product of the Golgi body, transported to the plasmalemma and cell wall through
pinocytosis. Callose represents a β-glucane, with increased permeability compared
to cellulose.
Physiological features: as a result of evolution, the root system in superior plants
has acquired the ability to perform growth movements towards water sources,
called hydrotropism.
Root orientation in space, its hydro-, chemo-, geotropic and branching behavior
is regulated by endogenous phytohormones (AIA and ABA).
Absorption of water and mineral salts is facilitated by the presence of the
rhyzosphere and symbiotic bacteria and fungi.
3.5 The Influence of External Factors on Water Absorption Intensity 49
Transpiration intensity represents the index reflecting the amount of water evap-
orated from a surface unit per one unit of time. This depends on the species, light
intensity, temperature, wind speed (15–250 g/m2/h during the day and 1–20 g/m2/h
during the night). During the 24 h cycle the intensity of the transpiration forms a
characteristic curve in which the minimum values are registered in the morning,
then they gradually increase at 1–2 PM (when they reach the maximum values), and
later decrease until 6 PM (when the intensity is very weak).
Transpiration productivity is the index reflecting the amount of organic mass
accumulated in plants during the elimination of a kilogram of water through tran-
spiration (1–8 g of dry matter per 1 kg of water).
The transpiration coefficient is the amount of water needed for the synthesis of a
gram of dry mass, it varies from 250–300 to 700–800 l/kg, depending on plant
species.
Relative transpiration is the index showing the water removal rate per unit of
leaf surface divided by the water evaporation rate from a free surface and has values
ranging from 0.1 to 0.5.
During the summer a corn plant eliminates through transpiration 150 g of water,
a pea plant −5 kg, 1 ha of oats—3,000 kg while cacti that grow in deserts eliminate
only 2,700 kg per ha annually.
The leaf is the basic organ where the process of photosynthesis takes place and is
the main body for water removal via transpiration. For creating a larger surface area
contacting with the atmospheric air in order to absorb more carbon dioxide (0.03 %)
and sunlight, plants have to develop a foliar system as large as possible, but this
creates a large surface of water evaporation as well. This is why, during evolution
plants have developed a morphological-anatomical structure to balance and opti-
mize both of the functions in order to survive (Fig. 3.5).
3.7 Structure of the Leaf as an Organ of Transpiration 51
Leafs have a thickness of about 100–200 µm. Parenchymal cells of the leaf are
arranged to form a sponge-like tissue. Between them there is a system of lacunae
(gaps) constituting 15–25 % of the leaf volume. Carbon dioxide diffuses through
assimilatory tissues, especially through the lacunar one (15–20 %). The leaf is
covered with a protective tissue—the unicellular epidermis, consisting of compact
cells. Each second cell in the leaf mesophyll is surrounded by a xylem vessel. Both
in the petiole and mesophyll water moves through the veins, whose number
decreases in the direction of the petiole. The smallest ribs consist of unique trac-
heids. In the leaves of some plants, especially those with C4 photosynthesis type,
the conducting vessels are covered with a compact layer of parenchymal cells
surrounding the vessels and serving as a mechanical support.
The leaves of certain species are covered with a cuticle containing oxymono-
carbonic acids impermeable to water. The cuticle contains waxes or hairs that
diminish water loss, reducing the speed of air movement and scattering the light.
Throughout the period of growth, plant transpiration at the leaf level happens in a
proportion of about 90 % through the stomata and 10 % through the cuticle.
Stomata are found on the epidermis of unsuberized leafs and stems (Fig. 3.6).
Stoma represent special pores in the epidermis of leaves and other organs which are
surrounded by two guard cells with a special shape (specialized parenchymal cells)
52 3 Water Regime
and are the basic structures participating in water removal. Guard cells are char-
acterized by:
• a prolonged shape in monocotyledonous plants and a semioval shape in
dicotyledonous plants;
• a thickened inner cell wall;
• chloroplasts containing chlorophyll.
Mature leaves can contain between 50 and 500 stomata. The majority of culti-
vated plants with the horizontal positioning of the leaves contain stomata only on
the bottom side of the leaf and are called hipostomatic. Plants with leaves posi-
tioned more or less vertically contain stomata on both sides of the leaf and are
called amphystomatic. Aquatic plants containing stomata on their top parts, are
called hyperstomatic.
In most of the plants cultivated in the temperate zones, guard cells are located in
the leaf epidermis. Plants growing in wet zones have the stomata exposed on cells
located outside of the epidermis to stimulate transpiration. In drought-resistant
plants stomata are buried in the leaf mesophyll. On average 10,000 stomata can be
found on 1 cm2, making up 1–2 % of the foliar surface.
Transpiration of branches and stems is very reduced compared to that of the
leaves and happens at the level of lenticels. Lenticels correspond to a lack of
cohesion between the suberized cells or to an absence of suberization. They make
only up to 2 % of the suberized surface (see Chap. 9—Elimination of substances in
plants).
Plants can also regulate transpiration intensity and the volume of eliminated
water. This regulation is carried out by the alteration of guard cell shape that
contributes to opening and closing of the stomatal pore (the osteole). The mecha-
nism of such changes in shape is based on cell turgidity variation.
The following types of deformations can be distinguished in stomata (Figs. 3.7
and 3.8):
3.8 Stomatal and Cuticular Transpiration 53
osteole opening and abscisic acid—its closure). The degree of water supply, the
deficit in relative humidity of the atmosphere, the wind speed etc. are important
factors as well.
Water absorption happens on the entire root system surface (total absorption), but
the process is much more active through the specialized root hairs (active
absorption). Enabling absorption are: imbibitions forces, the osmotic pressure, the
suction force of the cells, cohesion forces, etc.
The imbibition force of protoplasmic colloids is ubiquitous but has a most
important role in seed germination. The seedlings formed during embryo germi-
nation contain cells with no vacuoles, which do not develop a strong osmotic force
that would allow water absorption. Water penetration is conditioned by the
hydrophilic colloids from the protoplasm and from the seed storage tissue. Water
entering by means of imbibition is sufficient to determine rupture of the seed
tegument, seed germination and certain levels of seedling growth.
The osmotic force is developed by the osmotic pressure of the vacuolar sap in
root hairs and cortical parenchyma. Water is absorbed when the osmotic force of the
plants exceeds the retention of the water by the soil. The osmotic force value is
3.9 Water Absorption Mechanism and Ways of Its Circulation in Plants 55
exposed opposite to the xylemic vessels, and to the cells with suberized cell walls.
At the level of the Kaspari strips the flow of water reaches the xylem vessels only
trough the passage cells, forming a pressure pushing water into xylem vessels with
a force of 1.0–2.5 atm due to the free water potential. This force is called root
pressure. The action of the pressure contributes to water flow up the xylem vessels
when the foliar system is not present. This force is active and is based on the use of
metabolic energy (ATP).
An eloquent demonstration of the root pressure is the process of elimination of
crude sap from stems or sprouts when cut. (After the leaves unfold, stem injuries do
not cause sap elimination due to the suction force generated by the leaves.) In
perennial plants this phenomenon can be noticed mostly during the spring period
(birch juice) while for herbaceous—during the entire vegetation period. Crude sap,
released through this process, contains minerals, organic substances, sugars, etc.
3.9 Water Absorption Mechanism and Ways of Its Circulation in Plants 57
Summer crude sap in pumpkin plants contains ash particles, organic acids but no
sugar.
With the formation of the foliar system the importance of root pressure dimin-
ishes. During the warm summer period the essential role in water absorption is
played by transpiration. During water elimination the osmotic potential in meso-
phyll cells decreases, aspiring water from xylem vessels. A suction force of up to
30–35 atm forms in this case, contributing to water flow from conductive vessels to
the leaves and from roots—to xylem vessels. This force is called transpiration
pull, and is generated in the parenchymal cells of the leaf and has self-regulating
mechanisms. It is a passive force owed entirely to solar energy, which determines
the intensity of transpiration by heating the leaves. Due to this force, a continuous
flow is developing from the level of leaves to the root system level and during the
vegetation period plant water supply is ensured. Transpiration pull values are 15–20
times higher than the values of the root pressure (Fig. 3.11).
The distance water travels through leaf or root tissues is the fraction of a mil-
limeter, but it faces more resistance here, compared to when travelling through the
stem. Therefore, water flows through cortical and foliar parenchyma with a speed
10,000 times lower than the flow speed through xylem vessels.
Specialized elements for water transportation appeared in cormophytes—the
xylemic vessels, which supplied all plant organs and tissues. Xylem elements do
not contain cytoplasm and are formed from the procambial cells of the root and
stem. Cell walls rigidify and cells become dead to serve their role as conducting
tissue. Tracheids appear in pteridophytes and are composed of solitary cells with
perforated side walls. Tracheid appearance was an important step in the evolution of
the plant kingdom. Tracheids appear in gymnosperms and angiosperms and consist
of a large number of cells exposed in spirals, which have lost their lateral cell walls,
forming xylemic vessels.
Xylemic transport is obeying the laws of hydrodynamics by travelling along the
gradient of water potential from the root to the leafs. The active forces that
determine water circulation are created by the living cells at the upper and lower
Fig. 3.11 Water and nutrient circulation through the plant (John Johnson et al. 2002)
58 3 Water Regime
ends of the vascular system but a special role is also played by cohesion forces.
According to the theory of cohesion, water rises through the capillaries of the
conducting vessels owing to the transpiration pull and also due to the cohesion
forces between water molecules and the forces of cohesion of water molecules with
hydrophilic capillary walls (adhesion forces). The speed of water transport by
xylem vessels is 0.7–1.5 m/h in herbaceous plants and much higher in woody
plants. Cohesion force values can reach 300–350 atm.
When the stomata are closed and plants are supplied with water abundantly,
water elimination takes place due to the phenomenon of guttation, i.e. removal of
water drops by special structures called hydathodes. Guttation was observed in
more than 300 types of plants and is characteristic for tropical and equatorial plants.
Guttation is also an eloquent evidence of the root pressure.
The water regime is the totality of the processes of absorption, transport and
elimination of water, determining the ratio between the amount of water the plant
receives and the amount which is used by it in a time unit. The water regime reflects
the state of the water in the soil, the plant, the atmosphere and is in a dynamical
equilibrium. Water regime regulation contributes to the maintenance of the hydric
homeostasis.
Life had appeared in water and remained enclosed in the cells of living organ-
isms in a watery environment. During evolution, plants became more and more
independent from it, this fact allowing their expansion on the globe. Sporophyte
plants have retained their dependence on water in the reproductive process—their
gametes move through water drops and by using flagella. Gymnosperm and
angiosperm plants, by contrast, don’t need water for reproduction which is an
evolutionary advantage.
The ratio between the amount of absorbed and transpiration water characterizes
the water regime of different groups of plants and represents the water balance. In
gymnosperms and angiosperms this ratio is ≈1, due to:
• a perfectly developed root system, necessary for absorption;
• the conducting vessels, necessary for transport;
• the protective tissues, necessary to minimize evaporation;
• stomata, which regulate transpiration.
where
WB water balance,
A absorption,
T transpiration.
The water balance (WB) depends on the environmental conditions and deter-
mines the optimal functional activity of the plant. As a result of evolution, several
changes in organ structure and functions have emerged determining the water
balance and contributing to the formation of certain ecological groups of plants
characterized by specific water regimes. Terrestrial plants show a deficit of water
(5–10 %), which doesn’t lead to functional disorders. But a higher deficit causes
plants to lose their turgidity and wilt. Wilting may be temporary and permanent.
Plants have adapted to survive in conditions of water deficit, developing various
mechanisms to regulate water balance. Accordance to this, in 1973 Antipov pro-
poses a classification of plants (Fig. 3.12):
Fig. 3.12 The particularities of the water regime in different groups of plants
60 3 Water Regime
the transpiration area. The trees and shrubs from this group of plants have an
underground part often largely exceeding the size of the aerial one.
Hygrophytes are plants that grow in the regions with constant humidity. These
plants do not suffer from water deficit, so they have no special mechanisms that
would limit transpiration. They possess a specific type of parenchymal tissue—
aerenchyma, which stores oxygen needed for photosynthesis. The leaves are
hyperstomatic.
Mesophytes include most of the crop plants. The water regime in mesophytes
was studied by the Russian scientist B.D. Zalenski. He elaborated a law that bears
his name and states that leaves positioned higher on the stem possess better
expressed xeromorphic features—the cells are smaller, have a larger number of
stomata per surface unit of the leaf, the network of conducting vessels is denser, the
number of root hairs per surface unit is larger, the palisade tissue is better devel-
oped. Upper leaves are distinguished by a stronger assimilation and more intense
transpiration, the concentration of the cellular sap is higher and during wilting, they
remove the water from inferior leaves, leading to their drying and death.
One of the most important problems in biology is the problem of the evolution of
the organic world, i.e. the investigation of the ways and mechanisms that have
contributed to the formation of more complex structures and functions during
phylogenesis. Any function has evolved in parallel to its corresponding structure
and any structure—in parallel with its function—the evolution of photosynthesis,
transpiration, protein synthesis, etc. It is possible that the development of the water
regime has evolved with the emergence of different ecological groups of plants and
biological phenomena related to the general level of organization, for example,
plant adaptation to terrestrial life, the appearance of transpiration, the improvement
of the photosynthesis process, respiration, phytohormonal regulation, etc.; special
adaptive responses to one of the environmental factors, such as adaptation to
drought, to water excess, etc.
Based on this, there are hydrophytes (water-loving plants) xerophytes (resistant
to draught) and mesophytes.
It is considered that the evolution of superior plants has taken place at the
mesophyte level, characterized by the highest adaptability range to different envi-
ronmental factors.
In homeohydrophytes, during the ontogeny process, all types of water exchange
from the species phylogenesis appear, proving once more the universal law of
biology, the biogenetic law of Müller and Hegel—onthogenesis repeats the phy-
logenesis. From the zygote to the embryo stage the plant’s water regime is hyda-
tophyte. Then, from the seed formation to germination—the regime is
poichilohydric. During vegetative growth plants develop certain features that allow
strict regulation of the water balance and adopt the homeohydric water regime.
62 3 Water Regime
Glossary
Bound water Water linked with hydrophilic colloids of the protoplasm after their
hydration, being retained with big forces, doesn’t diffuse, freezes at temperatures
lower than −10 °C, doesn’t take part in the transformation and circulation of
substances.
Free water Water that preserves all the properties of pure water, moves freely, is
retained by relatively small forces, has solvent properties, evaporates via tran-
spiration and freezes at temperatures higher than −10 °C.
Water balance of plants The ratio between the amount of absorbed water and the
amount of water eliminated through transpiration. The value of this ratio
depends on environmental factors, A/T > 1 is characteristic for humidity excess,
and A/T < 1—for drought conditions. The volumes of transpired and absorbed
water should be equal for normal growth and development.
Wilting coefficient The amount of water in the soil expressed as a percentage, that
has remained unused by plants during their wilting. For sandy soils wilting
coefficient is 0.9 % and for the clayey ones—9.7 %.
Transpiration coefficient The amount of water (g) eliminated by plants through
transpiration, necessary for accumulation of 1 g of dry matter. Usually varies
from species to species within the limits 300–1,000.
Cohesion Property of water molecules to remain united due to attraction forces
(hydrogen bonds). This phenomena can be observed in xylem tissues enabling
water circulation in plants.
Hydropassive stomata movement Osteole closure in conditions of high humidity.
Takes place when guard cells are being pressed by the surrounding tissue at high
water concentration in leaves (for example, during the long-periods of rain).
Stomata open passively when weather is stabilized again.
Osmosis Diffusion of water from a higher water potential to a lower one through
semipermeable membranes.
Root pressure The force that causes unilateral (upward) water movement through
the root vessels.
Transpiration productivity The value, which indicates the amount of dry matter
(g) accumulated by the plant during the evaporation of 1 kg of water through
transpiration. The average value of this index is 2–8 g.
Dead water reserve of the soil The amount of water absolutely inaccessible to
plants consisting of pellicular, hygroscopic and chemically bound water. The
dead water amount depends on the soil type and its mechanical composition. The
dead water content of fine sand is 1.3 %, of the sandy-clayey soil—10.2 %, of
the silty-clayey soil—14.5 %.
3.10 Ecology of the Water Regime in Plants 63
Crude sap Liquid eliminated by plants from the injured tissues of the stem or the
root under the action of the root pressure. Chemically, the sap is an aqueous
solution containing minerals and organic substances.
Turgidity Water saturation state of the cells. Such a state provides the mechanical
rigidity and strength of the tissues, contributing to plant shape maintenance and
orientation of plant organs in space.
References
proton gradient into ATP and NADPH+H+, water photolysis and O2 release. In the
dark phase, fixation of CO2 by ribulose-1,5-diphosphate happens mediated by the
enzyme RUBISCO, carbohydrate synthesis, with consumption of the ATP and
NADPH+H+ formed during the light phase. In parallel with photosynthesis a
process called photorespiration occurs characterized by CO2 elimination and O2
absorption. It is known to intensify during intense illumination, high temperatures
or low CO2 concentration.
Historical Background
1771—J. Priestley has demonstrated that O2 is consumed by animals and is pro-
duced by plants.
1779—J. Ingenhousz has shown that light is necessary for green plants to produce
oxygen.
1818—J. Pelletier and J. Caventou extracted a pigment from green leaves and
called it chlorophyll.
1840—J.B. Bousingault proposed a global reaction for photosynthesis.
1845—J.R. Mayer showed that solar energy is transformed in the energy of
chemical bonds.
1875—K.A. Timireazev formulated the idea of the global role of green plants.
4 Photosynthesis 67
Brief Updates
Solar energy accumulation during photosynthesis depends to a big extent on the
level of cell protection against oxidative degradation. Multiple antioxidant com-
ponents (omega-3 fatty acids, vitamin E, carotenoids, etc.)—substances, which act
to neutralize free radicals—protect both vegetal and animal cells preventing
apoptosis, because the fundamental cellular signaling processes and the mecha-
nisms of protection and adaptation are very conservative throughout the entire
living world. Multiple extracts of algae and higher plants, products of photosyn-
thesis, are capable to manipulate signaling processes in human cells and as a result,
gene expression. For example, phytoestrogens (a group of flavones that play the
role of messengers in plant—microbe interaction) mimic the activity of the human
hormone estrogen. Carotenoids, such as zeaxanthin and lutein protect the photo-
synthetic systems from the destructive action of ultraviolet light, but can also be
found in the human retina (Lutein is apparently employed by animals as an anti-
oxidant and for blue light absorption while zeaxanthin may serve as a photopro-
tectant for retina from the damaging effects of free radicals produced by blue light).
An important role in the regulation of plant life is played by the solar light spec-
trum, which, besides photosynthesis, participates in the regulation of certain
physiological processes (like growth and development).
68 4 Photosynthesis
hv
CO2 þ 2H2 S !½CH2 O þ H2 O þ S2
Photosynthesis takes place in all plant cells that contain green pigments (leaves,
branches, young stems, sepals, unripe fruits), but the organ specialized in fulfilling
this function is the leaf, which shows some features formed during a long process of
adaptation and improvement:
• a large, flat area, adapted for absorption of large amounts of solar energy and
CO2 from the atmosphere;
• an epidermis provided with stomata through which gas exchange and transpi-
ration occurs. Depending on the positioning of stomata in plants, they can be
divided in 2 groups: amphistomatic (with stomata present on both sides of the
leaf) and epistomatic (with stomata located only on one side of the leaf);
• the presence of organelles specialized for photosynthesis—chloroplasts;
• a bilayer structure, the assimilatory parenchyma being differentiated in palisade
parenchyma that plays the main photosynthetic role and spongy parenchyma with
a pronounced role in gas exchange. In young branches, seeds and unripe fruits,
assimilatory cells with chloroplasts are located in the parenchymal layers under
the epidermis. Intercellular spaces are very small which causes reduced CO2
absorption from the external environment (in comparison with green leaves);
• the presence of conducting channels (phloem and xylem) which deliver mineral
compounds and water to mesophyll cells and transport the elaborated sap with
synthesized organic compounds to all plant organs.
During evolution leaves have changed generating a great diversity, determined
by the structural changes adopted for carbon assimilation (Fig. 4.2). Some plants
4.2 The Leaf as a Specialized Photosynthesis Organ 71
Fig. 4.2 Structure of the leaf blade in plants with C3 and C4 types of photosynthesis
originating from tropical and subtropical zones (corn, sugar cane, etc.) have leaves
with a particular anatomical structure that differs from the leaves of plants growing
in temperate climate (300.000 species of plants), adapted to carry out photosyn-
thesis in certain environmental conditions. The leaves of these species are well
vascularized, the mesophyll is homogenous containing granal chloroplasts while
conducting vessels are surrounded by a compact layer of parenchymal cells,
forming a sheath of perivascular assimilatory tissue with big agranal chloroplasts
(Fig. 4.2). Perivascular sheath cells are separated from the mesophyll and from the
air of intercellular spaces by a film, resistant to carbon dioxide diffusion.
Adaptive changes developed for the fulfillment of photosynthetic functions have
been directed both towards ensuring the optimal conditions for intense absorption of
solar radiation and to protect cells from photooxidation caused by visible spectrum
radiation and UV rays. Depending on environmental conditions, the cell size, the
morphology of the assimilatory tissue, the content and ratio of basic pigments
(chlorophylls and carotenoids) change, allowing photosynthesis to proceed in con-
ditions of both strong radiation (for some desert species), and low light (for tropical
species living in the shade). In some species, the cells of the superior epidermis of the
leaves, can focus light due to their shape increasing its intensity by 15–20 times.
Leaves of the plants from sunny zones, have a small area, are thick, have a larger
number of stomata and long palisade cells with chloroplasts containing less chlo-
rophyll, but assimilating carbon more efficiently. Another measure to protect the
cellular structures from optical radiation consists in the synthesis of auxiliary
pigments with photoprotective properties. Such substances are anthocyanins,
present in higher concentrations in young and senile plants; they are often formed as
a result of plant response to a high intensity of the visible light, to ultraviolet
radiation, to low or high temperatures and to other stress factors. These red pig-
ments are located in the cells of the superior epidermis and provide an effective
screening in the green region of the spectrum in which the leaves are mostly
“transparent”. On the action of UV radiation, the synthesis of several phenolic
compounds is induced. They are accumulating in the cuticle and epidermal cells,
ensuring UV absorption and tissue protection from its damaging effect.
Other compounds playing the role of photoprotection in foliar tissues
are carotenoids, which ensure, at low concentrations, a strong absorption in the
72 4 Photosynthesis
Chloroplasts are organelles specialized for fulfilling the photosynthetic function and
represent microstructures with the length of 5–10 µm and a diameter of 2–3 µm,
with spherical, oval, discoid or ellipsoid shape. In the majority of green plants
ellipsoid chloroplasts predominate; this shape proved to be the most rational,
developing during the evolution of the vegetal world.
The number of chloroplasts varies from 20 to 100 per cell, depending on the
species, environmental conditions, foliar tissue. The plastids of the cell are con-
stantly moving, either passively with the cytoplasmic flow or actively, requiring
energy consumption and being determined by light intensity and by other factors.
The structure of chloroplasts. The fundamental substance of the chloroplasts,
called stroma, is limited to the exterior by a double lipoprotein membrane (with the
thickness of 10–30 nm) containing a large number of pores with a surface area of
30–40 nm2.
The internal membrane, that has no pores, is less permeable in comparison to the
external one, but it can be passed by molecules of trioses and amino acids. It forms
folds called thylakoids (from thylacoides—“bag-shaped”) along the longitudinal axis
of the chloroplast and which either have the form of overlayed disks (these are called
granal thylacoids) forming the structures called grana (from granum—“granule”)
or traverse the chloroplast from one edge to another—thylakoids of the stroma
(Fig. 4.3).
The coordinated activity of the nuclear and chloroplast genomes in the synthesis
of these molecular complexes is represented schematically in Fig. 4.5.
Another example of regulation of the photosynthetic apparatus by the nuclear
genome is the synthesis of the key enzyme of the dark phase (the Calvin-Benson
cycle)—ribulose-1,5-bisphosphate carboxylase (RUBISCO). The functionally
active enzyme is composed of eight small and eight large subunits (Fig. 4.6). The
bigger subunit (54 kDa) is encoded by the chloroplast DNA, and the smallest
(14 kDa)—by the nuclear DNA, which, after being synthesized (translated) in the
cytoplasm, is transported into chloroplasts, where the correct assembly of the
Fig. 4.5 The assembly of the basic ETC complexes, formed by proteins encoded by the nucleus
(yellow) and the chloroplast (green) (Hermann 1992). Violet color shows unknown origin. 1 PS II;
2 cytochrome complex b/f; 3 PS I; 4 ATPase complex
enzyme takes place in the presence of another chaperone protein of 60 kDa also
encoded by the nucleus.
Proteins encoded by the nucleus with chloroplast destination enter the organelle
using a localization signal (a peptide of about 40 aminoacids for stromal proteins
and more than 80 for thylacoidal proteins localized at the N-terminus of the protein
molecule. This region recognizes the receptors of the external membrane of the
chloroplast, triggering the translocation of the entire protein into the stroma, where,
under the action of a specific peptidase (signal peptidase) cuts the localization signal
from the protein (by hydrolyzing a peptide bond). The protein can then be included
in the corresponding poly-enzymatic complex.
The biosynthesis of proteins encoded by genes of the chloroplast genome occurs
in the stroma—the fundamental substance of the organelle, which contains ribo-
somes, RNA, DNA and other components of the proteosynthetic system. Proteins,
specific for thylacoidal membranes are distributed according to certain physical-
chemical factors, including the amount of charge on the membrane surface. Big
differences have been noticed between the localization and the ratio of protein
complexes within the thylakoids of the grana and that of the stroma. Photosystem I
and the H+-ATPase complex are located in stromal thylakoids, while photosystem
II and the proton pumps—in granal thylakoids.
The physiological activity of the chloroplast is ensured by a certain level of
hydration (on average 58–75 %) and by a whole range of organic and mineral
substances present in the stroma, in which the enzymatic reactions of photosyn-
thesis take place (the dark phase). Chloroplasts contain enzymes involved in the
process of photosynthesis, carbohydrate phospholipid and chlorophyll biosynthesis,
in reactions of chlorophyll and starch degradation etc. Some enzyme molecules are
absorbed by the chloroplast lamellas, others can be found in free state. The largest
amounts of chlorophyll and carotenoids are concentrated in thylakoids (Table 4.1).
The formation of all molecular and structural components which enable the
physiological role of the chloroplasts (Fig. 4.7) is ensured by the nucleus-cyto-
plasm-chloroplast interaction which explains the failure to obtain isolated cultures
of chloroplasts on a nutrient substrate.
The origin of chloroplasts. Chloroplasts represent a variety of the organelles
specific for plant cells—the plastids, formed from the so-called proplastids, found in
meristematic cells. Proplastids are colorless vesicles with a diameter of 0.5–1.5 µm,
delimited by a double lipoproteic membrane containing a double stranded circular
Fig. 4.7 The schematic representation of the physiological role of the chloroplast
coiled DNA molecule with a size of 130–160 thousand base pairs (bp) encoding
about 130 genes. Proplastids have no developed internal membrane system.
The cytoplasm of the mother cell contains some proplastids, which pass into the
daughter cell during division, these proplastids in turn divide to increase their
number. Thus, genetic characteristics determined by the chloroplast DNA are
transmitted only through the maternal line (cytoplasmic inheritance).
Other types of plastids from plant cells, which may be colorless (leucoplasts,
etioplasts) or colored (chloroplasts, chromoplasts) can be also formed from
proplastids.
With cell growth proplastids increase their size as well, the internal membrane is
growing more intense, forming vesicles (thylakoids), which lie parallel along the
stroma. In the presence of light, these discs arrange in a certain position relative to
the plastid axis, suffering a differentiation into stromal and granal thylacoids. This
process is accompanied by the biosynthesis of lipids, proteins, chlorophyll and their
inclusion in the structure of the membranes. In the dark, etioplasts form, which have
an internal structure similar to a crystalline network of protochlorophyllids (chlo-
rophyll a precursors lacking the phytol side chain, which can be added only in the
presence of light in angiosperms). Ethyolated tissue exposure to light causes the
reorganization of the etioplast internal structure into a membranous structure,
characteristic for chloroplasts (Fig. 4.8).
Under the influence of certain environmental factors as well as according to a
specific genetic program, chloroplasts can transform into etioplasts, amyloplasts
(plastids that deposit starch) and chromoplasts (that are forming during the
autumnal period in leaves and some fruits and flowers during their maturation,
giving them the characteristic yellow-orange color, determined by the carotenoid
pigments). Chloroplast transformation into chromoplasts is caused by the destruc-
tion of the granal thylakoide structure and the formation of a new internal structure
4.3 The Structure, Chemical Composition, Function and Origin of Chloroplasts 77
under the control of the nucleus through specific proteins encoded by it and syn-
thesized in the cytoplasm. For example, the 58 kDa polypeptide that forms com-
plexes with carotenoids represents half of all the proteins contained in the
membrane structures of the chromoplast.
The differentiation of plastids from proplastids into green photosynthesizing
chloroplasts, into white amyloplasts containing starch or into yellow-orange chro-
moplasts, full of carotenoids and their inter-conversions are controlled by their own
DNA and the nucleus-cytoplasm interaction.
The study of molecular mechanisms of plastid differentiation and transformation
represents a big interest to modern molecular biology, because this knowledge
allows to understand better the signaling systems, which ensure the coordinated
functioning of the nuclear and chloroplast genome in the plant cell.
The selective absorption of solar radiation in the visible spectrum region of 400–
700 nm at levels sufficient for photosynthesis is a particular feature of several
organic compounds (pigments), which have in their structure chromophore groups
and systems of conjugated bonds. Plastid pigments may have a different chemical
composition and are classified into three groups: chlorophylls, carotenoids and
phycobilins.
78 4 Photosynthesis
Chlorophyll pigments are present in superior plants and in algae in four forms:
chlorophyll a”, “b”, “c” and “d”. All photosynthesizing plants and all groups of
algae and cyanobacteria contain chlorophyll “a”. Chlorophyll “b” is typical for
superior plants and green algae. Brown algae contain chlorophyll “c” and red algae
—chlorophyll “d”. Photosynthesizing bacteria contain various types of bacterio-
chlorophyll (Fig. 4.9). In the seeds of certain plants (pumpkin, hemp) protochlo-
rophyll was detected. In superior plants this chlorophyll type is synthesized during
the dark phase and on light exposure transforms into chlorophyll.
Chlorophyll is one of the most complex organic substances, representing a
double ester of chlorophyllin with phytol (the alcohol of a higher carbohydrate with
20 carbon atoms and a double bond) and methyl alcohol. The most common are
chlorophyll “a” and “b” (Fig. 4.10).
The most important peculiarity of photosynthesis is the use of solar energy, that is
the energy of electromagnetic oscillations, characterized by a certain wavelength
(distance between two consecutive maximum points of a cycle), an oscillation
frequency and a speed of dispersion:
c
k¼
m
where
λ—wavelength, nm;
c—speed of light, 2.997 × 108 m/s;
ν—oscillation frequency, Hz.
Solar light includes radiation from:
The visible spectrum: violet (400–450 nm), indigo (400–450 nm), blue (450–
500 nm), green (500–570 nm), yellow (570–590 nm), orange (590–610 nm), red
(610–700 nm) as well as from the invisible one, which includes:
82 4 Photosynthesis
hc
E ¼ hm ¼
k
Table 4.2 Light energy Plant species Light utilization coefficient (%)
utilization coefficient
Corn 4.5
Barley 2.9–3.5
Oats 3.3
Spring wheat 3.3
Potato 3.0
Fall rye 2.6
Beet 2.1
magnesium in the porphyrin core. Chlorophylls absorb orange and yellow light very
weakly and do not absorb green and infrared light at all. The position of the
absorption maxima in the spectrum is influenced by the nature of solvents, the
extent of interaction between chlorophyll molecules with themselves and with other
pigments, lipids and proteins.
The absorption maxima for carotenoids lay in the violet-blue region of the
spectrum—400–500 nm (Fig. 4.17). Energy absorption in the blue-green region of
the spectrum, 70 % of the absorption is due to these pigments and only 30 %—due
to chlorophyll. This feature of carotenoid absorption is important in carrying the
process of photosynthesis during cloudy weather when blue-violet rays dominate,
because the molecules of carotenoids (in contrast to xanthophylls) function as
auxiliary pigments in transferring energy to chlorophyll molecules.
Chlorophylls absorb violet-blue light more intensively, than red light (Fig. 4.18)
but carbon assimilation in the red region of the spectrum is more efficient. The
intensity of the photosynthesis process in different regions of the spectrum is called
action spectrum. The action spectrum is similar to the absorption spectrum by the
position of the light absorption regions and photosynthesis but it reflects photo-
synthesis intensity values (rather than absorption values) which indicate the varying
efficiency of using energy of different wavelength absorbed during photosynthesis.
86 4 Photosynthesis
The processes occurring during the light phase of photosynthesis can be related to:
(1) Absorption of carbon dioxide;
(2) Absorption of solar energy and its transformation into chemical energy.
(1) Absorption of carbon dioxide from the external environment happens
through the open osteole (photoactive physiological reaction). Carbon dioxide
enters the sub substomatal cavity, from where it diffuses through the free inter-
cellular spaces to directly contact the cellulose membranes of palisade assimilatory
parenchyma, situated on the upper side of the leaf blade, or the cells of the spongy
parenchyma from the inferior side (Fig. 4.19).
88 4 Photosynthesis
Fig. 4.20 Energetic levels of the molecules and transitions between electronic states
Electrons rotate around the nucleus, but also around their own axis, creating
mechanical and magnetic states—the so-called electron spins. Spins of two elec-
trons occupying the same orbital are oriented anti-parallel. The basic energy state of
the electrons, called singlet S0 (their total spin equals zero) is established, when all
the electrons are coupled (in pairs of 2) and occupy the lowest energy orbitals.
During photon absorption, electrons jump to higher energy levels, determining
the appearance of two singlet states S1 and S2, if the electrons don’t change their
spin and triplet T1, if one of the electrons changes its spin.
The highest energy level is level two singlet (S2). At this level the electron comes
under the influence of violet-blue rays, whose quanta carry a big amount of energy. In
the first excitation state (S1), electrons can continue absorbing quanta of lower
energy. The duration of the S2 excitation state is very short (10−12 s), and is followed
by energy loss as heat and transfer of the electron without changing spin direction, to
an inferior energetic level of excitation—the level one singlet (S1). The electron can
stay in the S1-state for a longer period of time (10−9–10−8 s). The duration of the
excitation state is a thousand times longer at the triplet level (10−5–10−4 s), which
appears on energy level transformations accompanied by electron spin changing.
Only electrons in S1 and possibly T1 states participate in photochemical reac-
tions. From the excitation state, electrons go back to the fundamental state S0
through:
• transfer of electronic excitation energy to a neighboring molecule of pigment
(photochemical reaction);
• its elimination as heat, fluorescence or phosphorescence (when transitioning
from T1 to S0).
The wavelength of fluorescent light is higher than that of absorbed light, because
the amount of energy is lower compared to the absorbed energy.
In the chlorophyll molecule there are two excitation levels, which determine the
presence of two absorption peaks. The first excitation level is related to the electron
transfer to a superior energetic level in the system of conjugated double bonds
4.6 Photosynthesis Mechanism 91
(18 delocalized π-electrons); the second excitation level is related to the paired
electrons of the nitrogen and oxygen atoms from the porphyrin core.
As mentioned earlier not all chlorophyll molecules fulfill the function of pho-
tochemical energy transformation and those who do, absorb only one quantum of
light in 0.1 s and have a very short excitation period. Hence, the full use of energy
from light quanta is possible only with the participation of additional pigments that
absorb and transmit solar energy to the photochemically active molecules, thus
ensuring an efficient functioning of photosynthetic system even at low values of
light intensity.
When a light quantum is absorbed, chlorophyll or auxiliary pigments from the
antenna transit into the state of electronic excitation:
Chl þ hm ¼ Chl
The energy of the excited molecule Chl* is transferred to the neighboring pig-
ment, which transfers it, in turn, to other pigment molecules with higher and higher
absorption wavelengths characterized by a lower level of singlet excitation down to
P680 or P700:
Oxidation of the reaction center and stabilization of separated charges. The first
reactions following light absorption and chlorophyll excitation in the reaction
centers are processes involving electron transfer between different macromolecular
entities. The sequence of events occurring in the reaction center is similar in all
photosynthesizing systems. The first photochemical reaction in the reaction center
is the fast transfer of electrons (τ ≈ 10−12 s) from the photoactive pigment (P*) to
the primary acceptor (I), for instance bacteriopheophytin (BPP) in case of bacterial
photosynthesis, the monomeric form of chlorophyll (A0) or pheophytin (Phe) for
photosystem I and photosystem II, respectively (Table 4.4).
Finally, this process yields a reductant I− (donor of electrons) and a strong
oxidant P+ (acceptor of electrons):
hc þ PI ! P I ! Pþ I
This marks the second important stage of solar energy transformation within the
process of photosynthesis (charge separation in the reaction center).
The subsequent electron transfer happens outside the center of reaction, through
an electron transport chain (ETC), which unites both reaction centers by means of
cytochrome b6-f. The positive charge of the reaction center, formed during its
oxidation, is neutralized within 2 μs by acceptance of an electron, returning to its
fundamental reduced state. The function of the electron donor is performed by
cytochrome “c” in bacteria, by plastocyanine in PSI and by the tyrosine radical—a
component of the water dissociation system in the PS II.
Electron transfer mechanism. The electron is transferred from the reaction center
at long intermolecular distances from one side of the membrane to another at high
speed. For example, the electron is transferred from P* to QA, at a distance of about
50 Å in 150 ps (Fig. 4.23).
Fig. 4.23 Energetic levels of the photosystem at the different states of the: reaction center (P),
electron donor (D), primary acceptor (I) and secondary acceptors (QA and QB)
4.6 Photosynthesis Mechanism 93
Fig. 4.25 Location of electron transporting complexes (PS I, PS II and b/f) and their interaction in
thylakoid membranes
QH2 leaves the photosystem II and can easily move inside the thylacoid
membrane ensuring the connection of PSII with other transporters of the ETC.
Thus, the plastoquinone play the role of a mobile carrier of two electrons. The
plastoquinone molecule diffuses into the galactolipid layer towards the complex of
cytochromes b/f with which it connects by yielding 2ē to it. For each molecule of
plastoquinone oxidized by the cytochrome complex, two hydrogen ions are elim-
inated inside the thylacoid (QH2↔Q + 2ē + 2H+). The oxidized plastoquinone
returns back to its location in order to make a new transport of protons and elec-
trons. The b/f complex serves as an electron donor for plastocyanin—a relatively
small hydrosoluble protein (the redox reactions of the plastocyanin are accompa-
nied by a change in the valence of copper ions, Cu2++ē ↔ Cu+). The molecules of
plastocyanin move easily along the lumen of the thylakoid and transport one
electron from the b/f/complex to PS I:
thylakoid membrane and when it receives two electrons it reduces the molecule of
NADP+ to NADPH+H+ in the presence of protons from the stroma.
Chlorophyll molecules in oxidized form Pþ 700 from the PS I reaction center are
reduced and return to their initial state by receiving an electron from PS II through
the reduced plastocyanin, which is being oxidized after releasing the electron.
Thus, as a result of simultaneous functioning of both photosystems, two elec-
trons from the molecule of water dissociated by the PS II are transported through
the ETC towards NADP, forming a strong reducing agent (NADPH2).
Photo-oxidation of water and elimination of molecular oxygen. The photosyn-
thetic oxidation of water is performed by the macromolecular complex PS II, which
includes three basic structural and functional parts:
• the complex of antenna-pigments, located on proteins with a molecular mass
ranging from 25,000 to 47,000;
• the reaction center with all its basic components located on a complex formed
by 2 proteins with a molecular mass of about 32,000, called D1 and D2 which
are located across the photosynthetic membrane (Fig. 4.26);
Fig. 4.26 Redox transformations and diagram of probable location of plastoquinone molecules in
the membrane (Tihonov 1996)
4.6 Photosynthesis Mechanism 97
The ions of manganese, when reaching a high level of oxidation, determine the
oxidation of water molecules:
The resulting electrons are taken by the manganese atoms which, thus, reduce.
The protons are accumulating in the lumen of the thylakoides while oxygen diffuses
in the cytoplasm, where it is dissolved in water, passes into the intercellular spaces
in the form of gas and is eliminated into the external environment through the open
stomatal pores. The tyrosine radical (Tyr Z)—the aromatic amino acid from the D1
protein (161st from the N-terminus) which plays the role of intermediary transporter
98 4 Photosynthesis
Fig. 4.27 Location of the basic components of the water photo-oxidation complex PS II (Klimov
1996). P680 Primary electron donor; Phe, QA, QB Electron acceptor; TyrZ Secondary electron
donor; D1 and D2 Proteins of PS II; cyt b559 Cytochrome with protective function by neutralizing
separate charges in cas of photoinactivation of PS II; 17, 24, 33, 43, 47 proteins with Mn (17,000,
24,000, 33,000, 43,000, 47,000 Da)
of electrons between Pþ 680 and the Mn complex or the radical of another aromatic
amino acid—histidine, also participate in the process of accumulating oxidative
equivalents. Also, it has been found that the ions of Ca2+, Cl− and possibly HCO3−
have a significant role in the functioning of the water oxidation system.
Conjugation of electron transport with proton transport and ATP synthesis. The
transfer of electrons at the level of mobile plastoquinone conjugated with trans-
membrane crossing of the H+ ions from the stroma to the inner space of the
thylakoids (against the concentration gradient) and the continuous influx of protons
generated by the enzymatic reactions of water photo-oxidation determine their
accumulation in the lumen of thylakoids. Due to the fact that the membrane of the
thylakoids is impermeable to protons, their concentration inside the thylakoids
increases a 100–1000 times in comparison with the stroma, causing the appearance
of a gradient of protons ΔpH and of a membrane potential Δφ conditioned by the
appearance of a positive charge inside and of a negative charge outside it. The
electro-chemical potential of hydrogen ions ΔµH+ (electrical potential—Δφ and
chemical potential ΔpH) is the driving force for the process of phosphorylation. In
4.6 Photosynthesis Mechanism 99
Acyclic photophosphorylation is specific only for green plants and for algae.
It has been noticed that during the light phase of the photosynthesis the quantity
of the synthesized ATP is higher than that of NADPH+H+. This surplus of ATP is
100 4 Photosynthesis
formed only in one photosystem—PS I. In this case, the transfer of electrons by the
transporters is cyclic (closed) and only includes the acceptors from the reaction
center (A0, A1, Fe-S proteins, Fd), the mobile plastoquinone, the complex of
cytochromes b6/f and the plastocyanin. The electron of the excited molecule from
the reaction center is successively accepted by both the primary and the secondary
acceptor until it reaches Fd. From there, it is accepted by PQ, which performs the
transfer of the electrons conjugated with the transmembrane transfer of protons
(from the stroma into the thylacoid cavity), on the cytochrome complex b6/f from
where it is accepted by Pc, which is passing it to the oxidized reaction center Pþ 700
reducing it. In this case NADP+ is not reduced, and the electro-chemical potential
generated by this electron transfer provides the phosphorylation of ADP (Fig. 4.29).
The summary equation of the cyclic phosphorylation is:
ADP þ Pi þ hm ¼ ATP þ H2 O
The final products of the light phase are O2, ATP and NADPH+H+. The later are
used, at a rate of 3/2, in the second phase of photosynthesis—the enzymatic phase
of carbon assimilation by reducing it to primary sugars (CH2O). This phase of the
photosynthesis takes place in the stroma of the chloroplasts. The surplus of ATP
produced during photosynthesis is used in other processes occurring in chloroplasts
(synthesis of fatty acids, of certain amino acids, reduction of nitrites etc.).
There are different methods of reducing carbon dioxide: the Benson-Calvin cycle
(C3), the Hatch-Slack-Karpilov cycle (C4), the metabolism of organic acids in
Crassulaceae (CAM—“crassulacean acid metabolism”) and photorespiration
(Fig. 4.30).
The Benson-Calvin cycle (pentose-phosphate reduction pathway, photosynthetic
type C3) is specific for a group of superior plants and includes a cycle of enzymatic
reactions that can be grouped in 3 main stages: carboxylation, reduction and
regeneration.
The carboxylation phase, the primary CO2 acceptor is a compound with 5
carbon atoms, ribulose-1,5-diphosphate, which forms as a result of secondary
phosphorylation of ribulose-5-phosphate with the participation of ATP and of
ribulose phosphate kinase.
Under the action of ribulose phosphate carboxylase/oxygenase (RUBISCO),
ribulose-1,5-diphosphate attaches a molecule of CO2 to the second carbon atom and
one molecule of water forming an instable compound with 6 atoms of carbon,
which splits into 2 molecules of 3-phosphoglyceric acid. RUBISCO, the key-
enzyme of the photosynthesis processes is the most widely spread enzyme on earth.
It is considered that the general quantity of this enzyme is 10 million tones or
approximately 20 kg per human being.
Fig. 4.31 Identification of the large (a) and small (b) subunits of the enzyme ribulose-1,5-
diphosphate carboxylase with the indication of the amino acid sequence. M Protein markers with
known molecular weights; [1–3] proteins extracted from leaves of different genotypes of
sunflower; [4, 5] proteins extracted from calathidia of different genotypes of sunflower
4.6 Photosynthesis Mechanism 103
formed as a result of CO2 reduction, consist of 4 carbon atoms. Due to this fact, this
type of carbon assimilation is called the photosynthetic type C4. This photosyn-
thetic type, is common for more than 1000 species originating from the tropical
areas, which are adapted to conditions of intense illumination and high temperature.
Plants in which photosynthesis takes place according to the C4 cycle, have
leaves with a particular anatomic structure (Fig. 4.33).
The cells of the palisade mesophyll have a small number of chloroplasts with CO2
fixation role, while the cells of the perivascular sheath are reach in big chloroplasts
and have the function of performing photosynthesis of the C4 type. Fixation of the
carbon dioxide takes place in the cytoplasm of mesophyll cells of, through a car-
boxylation reaction of the phosphoenolpyruvate in the presence of phosphoenol-
pyruvate carboxylase, resulting in a compound with 4 atoms of carbon—oxaloacetic
acid. In chloroplasts, oxaloacetic acid in the presence of NADP+H+ formed during
4.6 Photosynthesis Mechanism 105
the light phase and of NADP-malate dehydrogenase is reduced to malic acid. In the
presence of NH4+ ions the oxaloacetic acid can be aminated resulting in aspartic acid.
The malate (or aspartate) is transported through the plasmodesms from the mesophyll
cells to the cells of the perivascular sheath, which are permeable to organic acids and
impermeable to CO2. Here, it is decarboxylated, with the formation of pyruvate and
CO2. The pyruvate from the perivascular sheath is transported back to the chloro-
plasts of mesophyll cells where it undergoes phosphorylation in the presence of ATP
and phosphopyruvate synthase, thus regenerating the primary acceptor—the phos-
phoenolpyruvic acid.
In the chloroplasts of the perivascular sheath, PS II is weakly developed, in
comparison to PS I. The ATP necessary to fix CO2 is synthesized as a result of the
cyclic transport of electrons while NADP + H+ is formed as a product of the
oxidative decarboxylation of the malate concomitantly with CO2.
Due to the fact that two types of cells with two types of chloroplasts participate
in this mechanism of photosynthesis, this particular type of photosynthesis is seen
as “cooperative photosynthesis” (Karpilov 1970).
Fixation of CO2 via C4 has some advantages:
• The phosphoenolpyruvate carboxylase enzyme has a reaction speed higher than
that of ribulose 1,5-diphosphate carboxylase and this fact determines the
accumulation of carbon dioxide in the cells of the perivascular sheath.
• Some species can perform the first stages in which the organic acids are formed
during the night while, during the day, CO2 is released with subsequent re-
assimilation in the Calvin cycle. This fact allows plants to carry some carbon
assimilation reactions during the day, even when the stomatal pores are closed
(in the absence of exogenous CO2), thus, avoiding strong water elimination. It is
considered that such peculiarities form the basis of a higher drought resistance
of this group of plants;
• Carbon dioxide accumulation in the cells of the perivascular sheath, where the
C4 type photosynthetic reaction takes place determines the stimulation of this
process and concomitantly blocks the oxidase activity of ribulose 1,5-diphos-
phate carboxylase and, respectively, photorespiration. Thus, the unnecessary
consumption of organic substances is reduced and the productivity of the plants
is increased.
Crassulacean acids metabolism (CAM). Plants from the Crassulaceae,
Liliaceae, Cactaceae families and some of the Compositae, Osteraceae species etc.
have adapted to carry photosynthesis even in regions with drought and in deserts.
These plants fixate carbon dioxide in the same manner in which C4 plants do. But
for them two stage photosynthesis is specific: one during the night and another
during the day. The stomata of these plants are open particularly during the night
when most of the gas exchange processes happen. During daylight, they close
reducing thus the loss of water through stomatal transpiration.
The reaction of CO2 fixation by the phosphoenolpyruvic acid (first carboxylation
reaction) takes place in the cytoplasm in the presence of phosphoenolpyruvate car-
boxylase. As a result of this reaction oxaloacetate forms which, under the action of
106 4 Photosynthesis
Fig. 4.34 Comparison between C4 and CAM photosynthesis. a Spatial separation of the stages,
b temporal separation of the stages
4.7 Photorespiration 107
4.7 Photorespiration
protected in this manner by keeping the chloroplasts active. Such situations occur
during drought conditions, which make the stomata close. According to this
hypothesis, photorespiration which is useful in drought conditions becomes a
parasitic process under optimal conditions of life.
Fig. 4.37 Chloroplast development and interaction of chloroplast genes with nuclear genes
(Shestakov 1998)
membranes and the photosynthetic apparatus. Also, during the same period, fol-
lowing certain signals received from the chloroplasts, several genes that determine
synthesis of proteins destined for these organelles are activated in the nucleus. For
example, the proteins of the light-harvesting complexes which are involved in the
absorption and transfer of energy towards the reaction centre and those which par-
ticipate in the transfer of electrons from PS II to PS I are encoded only by nuclear
genes, synthesized in the cytoplasm and then transported to the chloroplast.
In the last stages of chloroplast development, self-assembly of the basic mac-
romolecular protein complexes of the photosynthetic apparatus takes place with a
maximum activity photosynthesis-related genes (Table 4.6).
Most of the genes from the chloroplast genome that encode photosynthetic
proteins become inactive in the mature phase of the organelle, except for those that
encode PS II proteins (e.g. th epsbA gene with D1 as its protein expression product)
(Table 4.7). At the same time, the level of expression of many nuclear genes
remains very high.
By applying molecular biology techniques, important information was obtained
regarding the structural organization of the photosystems, the cytochrome complex
b6/f, the ATP-synthase complex (Fig. 4.37; Table 4.7).
112 4 Photosynthesis
The synchronized activity of nuclear and chloroplast genes ensure the formation
of the structures and components of the photosystems, as well as of a number of
proteins that are not directly involved in electron transfer reactions, but still have an
important role in photosynthesis. These are the enzymes of chlorophyll and carot-
enoid pigment biosynthesis as well as proteases, kinases, phosphatases, translocases
with the function of transferring certain proteins from the cytoplasm to the chloro-
plast, metal-carrying proteins, ions, cofactors and other auxiliary proteins involved in
the assembly and renewal of the photosynthetic complexes, in metabolizing degraded
proteins etc. Mutations produced in genes that encode these proteins can cause a
decrease in photosynthesis efficiency or total inhibition of the process, similarly to the
effect of inactivating many genes that encode proteins of the photosystems.
The molecular mechanisms of photosynthetic gene regulation in chloroplasts are
different from those in the nucleus. Chloroplasts genes and ribosomes are similar in
4.8 Endogenous Regulatory Elements of Photosynthesis 113
structure to the prokaryotic genes and translation apparatus. Most of the chloroplast
genes lack introns, which are ubiquitous in eukaryotic genes. Also, many genes are
grouped in clusters, forming transcription units with promoters recognized by an
RNA polymerase of a prokaryotic type. In some photosynthetic genes, in the
promoter region, regulatory sequences with functions of transcription stimulation
and inhibition were identified. At the same time, it is known that the expression of
most chloroplast genes is subject to control after mRNA formation (post trans-
criptionally) but not at the transcription level.
Differential regulation of protein synthesis in the chloroplasts is determined by
mRNA transcribed with approximately the same efficiency from different genes, but
which are then subject to different levels of processing or degradation, depending
on the requirements for certain proteins (Fig. 4.38).
The stability of mRNA molecules also represents an important target for the reg-
ulation process. Some mRNAs are subjected to very fast degradation, others are stable
for a long time, participating in the synthesis of necessary proteins. mRNA stability is
determined by mRNA-specific proteins encoded by nuclear genes, which are located
in the 5′ untranslated regions. It is considered that for each mRNA there are specific
control proteins which determine their life span, processing and translation rates.
The expression of many nuclear genes, unlike those from the chloroplasts, is
effectively regulated at the level of transcription in the presence of specific regu-
latory proteins—photoreceptors. Both the phytochrome, which absorbs red light
with short and long wavelengths and some proteins which receives blue and
ultraviolet light have photoreceptor function. Phytochromes carry the specific
regulation of different groups of genes at different ontogenetic stages. The number
of nuclear genes whose expression is regulated by light by means of photoreceptors
is high, including the cab genes which encodes proteins of the light-harvesting
complexes. The activity of the cab genes is regulated not only by phytochromes but
also by phytohormones, by the redox-dependent systems of the cell. It is considered
that phytochromes act through other regulatory proteins—intermediate mediators
which are involved in the transmission of the light signal to genes controlling the
processes of photo morphogenesis and adaptation of the photosynthetic apparatus.
Membrane regulation of photosynthesis. Photosynthesis, like most of the
natural processes, is subject to feedback control, which implies that the speed of a
process which undergoes adjustment depends on its outcome. The electron transfer
speed in the ETC of a chloroplast or a mitochondria depends on the ratio between
the amount of substrate and the products of the ATP synthesis reaction
(ADP + Pi → ATP + H2O). Proton transfer processes conjugated with ATP syn-
thesis reactions play the key role in this regulation phenomenon.
ETC functioning determines the accumulation of hydrogen ions in the internal
space of the thylakoids, resulting in pH decrease. It has been proven experimentally
that illumination determines the decrease of chloroplast pH by 2.5 units. This
causes the speed of electron transfer to decrease. This reaction takes place at the
level of plastoquinone—an ETC step where electron transfer is accomplished the
slowest. Plastoquinone oxidation rates depend on the concentration of hydrogen
ions inside the thylakoids: the higher their concentration, the slower the oxidation
of QH2 occurs. This phenomenon is explained by the fact that electron transfer from
a reduced plastoquinone QH2 and its semi reduced form—plastosemiquinone QH∙
to the b/f complex (reactions 2 and 4) is preceded by proton dissociation states in
the internal space of the thylakoids—reactions 1 and 3 (Fig. 4.39).
The QH∙ and Q∙ forms are electron donors for the b/f complex. Dissociation
reactions 1 and 3 during which these active forms are produced depend on the pH
inside of the thylakoids. The lower the pH, the lower the probability of proton
dissociation, because the high proton pressure shifts the equilibrium of reactions 1
and 3 to the left in the direction of the synthesis of the inactive protonated forms
QH2 and of the plastosemiquinone QH∙. This is why accumulation of hydrogen ions
inside the thylakoids causes the decrease of the electron transfer rate.
The amount of ATP and ADP affects indirectly the electron transfer speed
through the ETC, at the level of proton transfer from thylakoids outwards through
the ATPase complex. The electron transfer speed stays at the highest rate as long as
substrate surplus exists: ADP and Pa (Fig. 4.39). Under these conditions, proton
transport through the ATP-synthase coupled with ATP synthesis occurs.
When ADP deficiency appears ATP synthesis literally stops. At the same time,
the transport of protons from the thylakoids into the stroma stops as well, but the
4.8 Endogenous Regulatory Elements of Photosynthesis 115
Fig. 4.39 Photosynthesis control in chloroplasts (Shestakov 1998). The chloroplast under
conditions of active ATP synthesis is shown at the top, while at the bottom—in conditions of
photosynthetic control where ATP synthesis is impaired. On the left the dependence of the rate of
plastoquinone oxidation by the b/f complex on illumination duration is shown
ETC continues to function (Fig. 4.39). Thus, proton transfer, combined with that of
the electrons determines the pH decrease, causing a slowdown in the oxidation
reactions of the reduced plastoquinone. Subsequently, these events slow down the
transfer of electrons between PS II and PS I. When chloroplast ATP reserves are
exhausted through their use in the Calvin cycle and other processes in the chlo-
roplast, ADP accumulation occurs and the same sequence of events repeats: excess
of ADP, ATP synthase stimulation, increase of the internal pH, intensification of
electron transfer.
Regulation of enzymatic activity through phosphorylation. To adjust the
distribution of solar energy in chloroplasts, besides the light-harvesting complexes
linked with PS I and PS II, there exists another complex of mobile pigments, which
acts as an additional antenna. PS I and PS II are not uniformly distributed in the
membranes of the granal and stromal thylacoids. Most of the PS I components are
located on stromal thylakoids, while PS II complexes—on granal thylakoids
(Fig. 4.40).
It is assumed that under low light conditions, the mobile antenna is localized
mainly in granal thylakoids near the PS II, due to which the size of the PS II
antenna increases. And under certain specific conditions, when the need to increase
the efficiency of PS I emerges, this mobile antenna complex leaves PS II, moving
through the membrane towards stromal thylakoids, where it contacts PS I. The
116 4 Photosynthesis
Fig. 4.40 Structure of the chloroplast ETC and the ATP-synthase complex (PQ Plastoquinone,
PC Plastocyanine, Fd Ferredoxin, Cyt Cytochrome) (Purves et al. 2005)
signal that determines the translocation of the mobile antenna is the surplus of
reduced transporters in the ETC chain between photosystems I and II. This surplus
may appear when PS II operates more intensely than PS I. Movement of the antenna
towards PS I determines the “unloading” of the ETC between the photosystems
based on more intense PS I functioning. This mechanism, which regulates the
movement of the mobile light collecting complex is determined by the phosphor-
ylation of a subunit of the protein complex through the attachment of a phosphoric
radical by a protein kinase. The phosphorylated, negatively charged complexes,
situated close to each other, reject reciprocally (rejection forces are of electrostatic
nature). Following such reorganization, of the light-harvesting complexes of PS II
decreases in size and that of the PS I increases, thus, ensuring a coordinated
operation of both the photosystems. This regulating mechanism allows the photo-
synthetic apparatus to react adequately to changes in lighting conditions. When the
need to expand the PS II antenna arises (in response to varying light intensity or
spectral composition), a protein phosphatase is activated which eliminates the
phosphate group by hydrolysis. The mobile dephosphorylated complexes move
again to the granal thylakoids in order to contact PS II. Thus, by means of protein
kinase and protein phosphatase action the chloroplast can optimize its power dis-
tribution between the light harvesting antennae of PS I and PS II.
Redox regulation of photosynthetic enzymes with thiol groups of protein
derived amino acids. Another mechanism of photosynthesis regulation includes
the redox transformations of the photosynthetic proteins. The carboxylation
4.8 Endogenous Regulatory Elements of Photosynthesis 117
reaction catalyzed by the RUBISCO enzyme is the slowest in the carbon fixation
cycle. The activity of this enzyme is controlled at the level of thiol groups, which if
in an oxidized state (S-S bridges) keep the enzyme idle and when reduced (–SH),
activate the enzyme.
An intermediate between this enzyme and the ETC, which is the source of
electrons to activate RUBISCO is a specific protein thioredoxin, which is wide-
spread not only in plants but also in the animal kingdom and bacteria. Thioredoxin
is subject to redox transformations in cells by reduction of the thiol groups
(-S-S + 2 ē + 2H+ → 2-SH). In chloroplasts, the thioredoxin is reduced by
receiving two electrons from two molecules of reduced Fd in the presence of a
specific enzyme ferredoxin-thioredoxin reductase. The reduced thioredoxin oxi-
dizes yielding the electrons to RUBISCO. Thus, transition from darkness to light
induces ETC activity, formation of reduced ferredoxin molecules and activation of
the RUBISCO enzyme (ferredoxin-thioredoxin-RUBISCO). Due to ribulose
diphosphate carboxylase activity, the speed of carbon dioxide utilization in the
Calvin cycle increases. Activation of this enzyme also depends on other factors:
changes in the pH and Mg2 ion content in the chloroplast stroma, that occur when
chloroplasts are exposed to light.
By exchanging substances and energy plants are in constant contact with the
environment. The ecology of photosynthesis involves the study of photosynthesis
productivity dependence on environmental factors—light, O2 and CO2 concentra-
tion, temperature, water, humidity, minerals (Mg, N, P, K), presence of toxic
substances which may inhibit photosynthesis etc.
The dependence of photosynthesis on light intensity and spectral composition.
Plant adaptation to different light intensities happened by means of different mor-
phological and physiological changes. If seedlings are exposed to darkness, chlo-
rophyll biosynthesis stops which determines the formation and accumulation of
colorless protochlorophyllides, thus resulting in etioplasts in which the process of
photosynthesis cannot take place. Exposure of etioplasts to light is accompanied by
structural and functional changes, which cause the photosynthesis process to
resume. Also, it induces differentiation of the internal membranes system as well as
synthesis of proteins and lipids. On average, leaves absorb 80–95 % of the pho-
tosynthetically active solar spectrum (at a wavelength of 400–700 nm) and 25 % of
the infrared energy, which constitutes about 55 % of the total radiation energy.
Photosynthetic efficiency and the direction of organic biosynthesis depend on the
quality of light absorbed by the leaf. The dependence curve of the photosynthesis
process on the intensity of light has a logarithmic shape. A directly proportional
dependence is observed only at low light intensities. Red light is always present in
direct sunlight. Plants that have been grown in blue and red light differ essentially
by the composition of photosynthesis products. In blue light other compounds are
118 4 Photosynthesis
• Autotrophic plants (from Greek autos “self” and trophe “nutrition”) pro-
duce by themselves all the necessary organic material using mineral sub-
stances absorbed from the external environment. Depending on the source
of energy they use, autotrophic plants are grouped in phototrophic plans,
which use solar energy, chemoautotrophic plants, which use chemical
energy produced by oxidation of certain mineral substances (H2S, NH3
etc.) in the process of chemosynthesis.
• The total amount of carbon, fixed by means of photosynthesis during the
entire year, is about 7.8 × 1010 tones. This quantity is compensated by the
same amount of CO2 which is eliminated through respiration by hetero-
trophic organisms. Assimilated carbon amounts to about ¼ of the total CO2
reserves in the atmosphere. Every year up to 0.3–0.4 % of the total reserve
of carbon from the hydrosphere and troposphere is assimilated.
• The total production of organic substance, synthesized by the vegetation of
the planet, calculated in glucose, sums up to 4.5 × 1011 tons per year.
• The world’s annual energy consumption (5 billion people) is 3 × 1020 J—
10 % of the energy accumulated annually in the process of photosynthesis.
About 1 billion tons is used in the form of food products solely. It equals to
120 4 Photosynthesis
(2 × 1020 kcal) or 1 over 15 billion of the total energy issued by the Sun.
Thus, out of the 40 % of the Solar energy that reaches the surface of the
Earth, 2–5 % is absorbed by plants, out of which only 0.1–0.22 % is used
in the production of organic matter.
Glossary
Carotenoids Yellow and orange pigments that are found in chloroplasts and
chromoplasts that participate in light absorption as supplementary pigments and
protect the molecules of chlorophyll and other active substances from irrevers-
ible photo-destruction. One can distinguish oxygen free carotenoids (C40H56-
lycopeneα-β-γ-carotenoids) and oxidized carotenoids (C40H56O2C40H56O4-
xanthophyllsluteinzeaxanthin).
Chloroplasts Specialized organelles of the vegetal cells in which photosynthesis
takes place delimited to the exterior by two membranes—internal and external
with the second being incorporated in the homogeneous environment (stroma).
The internal membrane form the folds called stromal thylacoids and granal
thylakoids, in which all the photochemical reactions of the light phase are
carried.
The dark phase of photosynthesis A complex process that includes the sequence
of enzymatic reactions that lead to the formation of photosynthesis products and
of the organic acceptor of carbon dioxide.
The light phase of photosynthesis A phase of photosynthesis during which light
absorption and transformation of solar energy into the chemical energy of ATP
and NADPH+H+ happens. This process occurs in the active photochemical
membranes of the chloroplast and represents a system of photophysicalphoto-
chemical and chemical reactions.
Acyclic phosphorylation A process during which light energy is transformed into
the macroergic bonds of ATP and NADPH+H+. It is then followed by water
photolysis and oxygen elimination.
Cyclic phosphorylation A process during which the electron emitted by chloro-
phyll through a series of transformations returns back to the pigment. The
absorbed energy is fixed in the macroergic ATP bonds.
Photosynthetic phosphorylation A process of converting light energy quanta into
ATP.
122 4 Photosynthesis
References
Historical Background
1770—A.L. Lavoisier for the first time emphasizes the role of oxygen in living
organisms.
1897—A.N. Bach formulated the peroxide theory of biological oxidation.
1912—V.I. Palladin proposed the idea of two stages in respiration: anaerobic and
aerobic.
1912—H. Wieland demonstrated the role of oxygen in oxidation processes.
1921—O.G. Varburg showed that oxygen assimilation is inhibited by carbon
dioxide.
1925—D. Keilin discovered cytochrome oxidase.
1935—G. Embden, O. Meyerhov and I. Parnas determined the most important
products of glycolysis.
1937—H.A. Krebs described the citric acid cycle (the Krebs cycle) in animals.
1939—A.C. Ibnell discovered the presence of the Krebs cycle in plants.
1939—H.M. Kalcar and B.A. Belitzer discovered oxidative phosphorylation.
1961—V.A. Engelhardt submitted the idea of oxidative phosphorylation and aer-
obic respiration.
Brief Updates
According to proteomic studies, each cellular body contains 1,000–2,000 different
polypeptides. Approximately 15 % of the cellular proteins are located in
5 Plant Respiration 125
Respiration is one of the basic characteristics of life and represents the physio-
logical process of oxidative degradation of complex organic substances to inter-
mediary products (fermentation) or to final products (aerobic respiration).
Aerobic respiration occurs in the presence of O2 and ends up with total substrate
degradation (catabolism) and energy release. Aerobic respiration happens in all the
cells at the mitochondrial level (Fig. 5.1).
C6 H12 O6 + 6O2 = 6CO2 + 6H2 O + 686kcal=mol 2867kj=mol
126 5 Plant Respiration
Anaerobic respiration occurs in the absence of oxygen and ends up with partial
energy release, which is characteristic of anaerobic organisms:
nðCO2 Þ 6ðCO2 Þ
CR ¼ ¼ ¼1
nðO2 Þ 6ðO2 Þ
When lipids and oxygen-poor proteins are degraded CR < 1 and, with its values
ranging between 0.3 and 0.5. For organic acids CR is ranging between 1 and 4. The
less oxygen is contained in the molecules of the respiratory substrate the more free
oxygen it will need during respiration, and the lower the CR value will be.
nðCO2 Þ 4ðCO2 Þ
2C2 H2 O4 ¼ 4CO2 þ 2H2 O þ Q CR ¼ ¼ ¼4
nðO2 Þ 1ðO2 Þ
nðCO2 Þ 18ðCO2 Þ
C18 H36 O2 þ 26O2 ¼ 8CO2 þ 18H2 O þ Q CR ¼ ¼ ¼ 0:69
nðO2 Þ 26ðO2 Þ
The respiratory index also differs depending on the age of the organ and of the
organism. Thus, in green fruits the respiratory index is 0.5–0.1, in mature fruits it
varies between 2.5 and 3.0, in flax seeds it is 0.65 and in tangerines it is 2.65. CR is
of crucial importance in the process of seed germination, as well as in regulating
fruit maturation and preservation. A low CR in oleaginous plants means that much
aeration is needed during germination. In tomatoes, blueberries, plums and melons,
by the end of their growth when organic acids are still being synthesized, CR is
0.85, during the aromatic phase in the same fruits it is 1.25, and during the maturity
phase CR is as high as 2.7.
128 5 Plant Respiration
Fig. 5.2 Activation energy with and without catalyst and the mechanism of enzyme action
this group of enzymes the most important one is succinate dehydrogenase, which
transfers electrons from aerobic dehydrogenases to quinones, cytochromes or O2.
Oxidases are enzymes which transport electrons and protons to the final
acceptor—O2. These enzymes are situated on the mitochondrial cristae and par-
ticipate in the redox reactions of the electron transport chain (ETC). Given their
action mechanism, it is considered that they participate in the process of activation
of atmospheric oxygen. Depending on the number of electrons transported, oxygen
can be turned into H2O, H2O2, or O2.
The cytochrome oxidases represent a coupling between A and a3 cytochromes.
Their structure looks like that of hemoglobin, because they are formed of a por-
phyrin core with four pyrrolic groups having iron as a cofactor in the center, which
during electron transport is oxidized and reduced successively:
The transport of electrons in the electron transport chain is carried out according
to the electric potential gradient. Electrons can be transferred to oxygen only by the
cytochrome-oxidase (A+a3) (Fig. 5.3).
Polyphenoloxidases contain copper atoms as cofactors. These enzymes partici-
pate in the oxidation of both polyphenols (respiratory chromogens) and quinones
(respiratory pigments).
Peroxidase and catalase, which contain the iron cation as a cofactor are part of
the oxidase group too. Peroxidase oxygenizes the organic substrate by transferring
protons to the peroxide oxygen resulting from oxygenated water decomposition:
Fig. 5.3 The path travelled by electrons through the cytochrome system down their way to
oxygen
Efforts made by the physiologists and biochemists at the end of the nineteenth
century contributed to the elucidation of the chemistry and mechanism of the
respiration process. Two theories related to the primary processes of substrate
degradation, are recognized:
• the theory of slow oxidation, defined in 1897 by A.N. Bach;
• the theory of respiration, defined in 1915 by V.I. Palladin.
According to the theory of slow oxidation, in aerobic respiration the main role
goes to oxygen which is activated by a easily oxidizable substance (A) from the
plant body. As a result of breaking the double bond between oxygen atoms and the
coordinative bond of –O–O type, peroxides are formed. This kind of reaction is
carried out with the help of peroxidase and cytochrome oxidase enzymes, which
together with O2 create peroxide combinations. This type of reactions is assisted by
enzymes of the peroxidase and cytochrome oxidase type which form peroxidic
combinations with O2. The oxidation cycle is repeated until the complete oxidation
of the substrate:
5.3 A.N. Bach’s and V.I. Palladin’s Theories 131
The modern concept concerning the chemistry of the respiration process has been
elaborated based on the works of the following scholars: A.N. Bach, V.I. Palladin,
O. Varburg, V.A. Enghelhardt, A.I. Oparin and others. It implies carrying out a
chain of redox reactions, catalyzed by specific enzymes. In case the respiratory
substrate is represented by soluble carbohydrates, the degradation reactions com-
prise two phases:
• anaerobic degradation;
• aerobic degradation.
Between 1912 and 1928 the physiologist S.P. Kosticev, continued to research the
mechanism of respiration in different living organisms and concluded that the initial
chemistry of the respiration process in plants, animals, human beings and micro-
organisms is identical. In other words the first stage of respiration, during which
132 5 Plant Respiration
GLUCOSE
Intermediate products
In conditions of
saturation with
Complete degradation of metabolic energy, AP Various types of
the substrate occurs in takes part in fermentation take place
aerobic conditions carbohydrate in anaerobic conditions
resynthesis
hexoses are decomposed anaerobically until pyruvic acid is formed, are based on
reactions identical for all living organisms.
The subsequent metabolic paths are different. In anaerobic conditions the
pyruvic acid is subjected to degradation through fermentation, while in aerobic
conditions—through the Krebs cycle. In cases when the cell does not require
energy, resynthesis of hexoses occurs.
Thus, Kosticev proved the genetic link between respiration and fermentation, at
the level of pyruvic acid. He also determined that this acid represents half of the
glucose molecule and is the fundamental plate of the cellular metabolism for both
catabolic and anabolic processes (Fig. 5.4).
Under anaerobic conditions, plants (germinated seeds, fruits and vegetables
stored in poorly ventilated warehouses, vegetal organs that are submerged in water
during floods or are under thick layers of snow) breathe through alcoholic fermen-
tation, but this process is inefficient from the energetic point of view and accumu-
lation of ethylic alcohol in plants can be toxic for the functioning of living cells.
Glycolysis, the first stage of the respiration process was discovered by G. Embden,
O.F. Meyerhof and I.O. Parnas (it is also called the EMP path, dichotomyic path,
anaerobic phase of respiration). It happens in the cellular hyaloplasm and in
chloroplasts. This stage involves endogenous oxygen and consists of three
5.4 Respiration Mechanism 133
successive phases during which the molecule of glucose is divided into two mol-
ecules of pyruvic acid:
• activation of the hexoses with the consumption of two molecules of ATP and
their dichotomic decomposition into two phosphotrioses—unstable intermediary
products (Fig. 5.5);
• the first substrate phosphorylation, which starts with the transformation of
phosphoglyceric aldehyde into phosphoglyceric acid with energy release in the
form of reduced NADH+H+ and synthesis of one molecule of ATP;
• the second substrate phosphorylation, during which the 3-phosphoglyceric acid,
as a result of intramolecular oxidation yields the phosphate while a molecule of
ATP is formed.
Glycolysis is energetically poor compared to oxidative phosphorylation. During
it, a macroergic bond is formed—the enolphosphoric bond (the highest-energy
phosphate bond found in living organisms, −61.9 kJ/mol), the energy of which is
used for synthesizing ATP:
General reaction:
The process ends up with the production of two molecules of pyruvic acid from
a molecule of glucose. Over time, two molecules of ATP are consumed and four
molecules of ATP and two of NADH+H+ are produced.
Importance of glycolysis
• Glycolysis is the initial and common phase of aerobic respiration and
fermentation.
• It is the link between the respiratory substrate and the Krebs cycle.
• It yields two molecules of ATP and two of NADH+H+ per each molecule
of hexose.
• A series of intermediary products which can be used in different metabolic
cycles are formed.
• In chloroplasts, it represents an independent way of ATP and NADH+H+
synthesis.
• By means of glycolysis, in this cellular organelles starch breaks down into
phosphotrioses—a compound that can be transported through the chlo-
roplast membrane.
The pyruvic acid that results from glycolysis is subjected to activation during which
an acetyl-CoA molecule is formed which goes into the Krebs cycle (tricarboxylic
acid cycle). These transformations, as well as the cycle itself occur in the mito-
chondrial matrix. The Krebs cycle is a succession of oxidative decarboxylation
reactions, of dehydrogenation reactions, water fixation or elimination reactions
which result in the formation of a series of intermediary products: citric
acid → isocitric acid → ketoglutaric acid → succinic acid → fumaric
acid → malic acid → oxaloacetic acid (the latter is converted into citric acid when
pyruvate is provided and the cycle restarts).
In the mitochondrial matrix different transformations occur that can be grouped
in 5 important phases (Fig. 5.7):
1. Oxidative decarboxylation of the pyruvic acid, along with substrate dehydro-
genation in the presence of the dehydrogenase, which transports the electrons
and protons to O2 resulting in water formation. Through acetylation acetyl
coenzyme A is formed (an important metabolic and energetic compound, which
includes a macroergic bond of 11 kcal) with the participation of thiamine
136 5 Plant Respiration
FAD
#
NADH ! FMN ! ubiquinone ! cytochrome b ! cytochrome c1 ! cytochrome c !
cytochrome a1 a3 þ 0:82V H2 O
" 1=2O2 þ 2Hþ
ATP biosynthesis happens only on the internal membrane. At this stage, water
forms from atmospheric oxygen and the hydrogen resulting from the oxidation of
NADH+H+, FADH+H+.
Electron transport is an exergonic process, which occurs with release of energy
and is powered by the difference of redox potential between the initial and final
systems. During transportation of a molecule of NADH+H+ and its oxidation in the
ETC, biosynthesis of up to three molecules of ATP occurs, while in case of oxi-
dation of one molecule of FADH+H+ or FMNH+H+, the biosynthesis of up to two
ATP molecules happens. Thus, the energetic balance of respiration can be repre-
sented as follows:
• glycolysis. 4 molecules of ATP result from the glycolysis process, out of which
2 are consumed for hexose activation through phosphorylation. In addition two
molecules of NADH are produced, which are transported to mitochondria (the
process of transport also consumes energy) and result in up to 6 molecules of
ATP in the ETC.
5.4 Respiration Mechanism 139
• pyruate decarboxylation and The Krebs cycle. 15 molecules of ATP for each
molecule of pyruvic acid and consequently, 30 molecules of ATP for each
molecule of glucose are produced. Out of these, 24 are incorporated in NADH
+H+ and, 6—in FADH+H+ while two molecules of GTP are obtained separately
during the cycle.
In sum 8ATP + 30ATP = 38ATP are produced per molecule of glucose. In reality,
this number is around 30–32 or lower, due to the need to transport compounds (e.g.
the pyruvate and the NADH produced during glycolysis, the phosphate, and the
ADP) across the membrane into the mitochondrion which requires energy, but also
due to the “leakiness” of the mitochondrial membrane for hydrogen protons, which
means that not all of the hydrogen protons are used for ATP synthesis.
By taking the ratio between the amount of energy released through respiration to
the amount of energy expended on anabolic processes the following energy effi-
ciency is obtained.
1.591 kJ:2.871 kJ
55.4 %
381 kcal:686 kcal
Fig. 5.8 Pentose phosphate pathway (F. Binet and J.P. Brunel)
Fig. 5.9 Glyoxylic acid cycle. Enzymes involved: 1 citrate synthase, 2 aconitase, 3 isocitrate
lyase, 4 malate synthase, 5 malate dehydrogenase
The internal factors that determine the intensity of plant respiration are:
• Temperature. Changes in temperature can produce certain changes in the
intensity of respiration, through the influence of temperature on the activity of
enzymes which act in biological oxidation processes. Temperature influences
the process of enzymatic glycolysis of glucose and the activity of enzymes that
activate hydrogen in the respiratory chain. At low temperatures, the respiration
process is catalyzed by flavoprotein enzymes, while at higher temperatures—by
oxidases, which play a more significant role. The different roles of these
enzymes in respiration depending on temperature demonstrates the adaptive
nature of the respiration process in plants.
• Light influences the formation of the organic respiratory substrates which are
consumed during the bio oxidative processes, thus, indirectly, it influences the
respiration process of green plants. The coenzymes involved in photosynthesis
(NAD, ATP, ADP) have also an important role in respiration. Thus, ADP is
essential for the terminal phases of oxidative phosphorylation, while the
reduction of NAD during photosynthesis inhibits the pentose phosphate cycle
and aerobic respiration.
• The influence of mechanical trauma, such as cutting or biting increases the
intensity of respiration, because some synthetic processes are intensified.
• Cytoplasm hydration degree. The substances which are decomposed in plants
during oxidation, are oxidized only if they are dissolved in water. This means
that the degree of protoplasm hydration influences the intensity of respiration. In
case of a prolonged water deficit, an essential change in the organic metabolism
can occur which can lead to a considerable drop in the crop yield.
• Presence of mineral salts in the soil (nitrates, nitrites, sulfates, ammonium and
potassium salts) intensifies respiration in vegetal tissues (anionic respiration). In
this case respiration intensity is directly proportional to the amount of absorbed
ions. The NH4+ ions participate in glutamine synthesis—a process during which
NADPH+H+ is formed. Potassium has a role in maintaining mitochondrial
structures. Sulfur is part of certain respiration enzymes (acetyl-CoA), iron is an
important element in the composition of cytochromes.
• Certain chemical substances can have an inhibiting action on respiratory
enzymes. Fluoride can stop glycolysis, while malonic acid can inhibit the Krebs
cycle. Cyanides influence the oxidative phosphorylation process as well as
electron transportation in mitochondria. Arsenates, 2,4-dinitrophenol break
down macroergic intermediary precursors, which precede ATP formation by
blocking oxidative phosphorylation, even though they stimulate electron
transport in the ETC.
• The chemical composition of the air (oxygen and carbon dioxide concentra-
tion) has a great influence on the respiration process. If the pressure of carbon
dioxide in the environment is high, than the intensity of respiration will grad-
ually decrease and the plant will absorb lower and lower quantities of oxygen.
144 5 Plant Respiration
Fig. 5.10 Carbohydrate degradation via respiration with CO2 release and ATP formation (Milica
et al. 1982)
5.7 Regulation and Self-regulation of the Respiration Process 145
Glossary
References
Historical Background
Brief Updates
In order to grow and develop, plants need a big number of chemical elements that
come either directly from minerals, or from mineralization of the decaying organic
substances in the environment. Mineral elements are required in different amounts
by plants. The amounts required differ from species to species and even from variety
to variety. In addition, plants have somewhat different specific requirements which
end up determining the chemical composition of different species.
Chemical analyses showed that the vegetal matter contains more than 60
chemical elements, but only 15 are necessary to provide the normal growth and
development of plants. The basic chemical elements: carbon(IV) and oxygen,
which are obtained from the atmosphere, hydrogen which is dissociated from water
and nitrogen—derived from mineral compounds are called organogenic elements,
since they represent the basic constituents of organic substances (amino acids,
proteins, carbohydrates etc.) and are most widely spread, representing 95 % of the
dry vegetal mass. The ions of nonmetals (N, P, S) and metal ions (Ca, K, Fe, Mg,
Cu, Mb, Zn) represent the key-components in the process of mineral nutrition and
are absolutely necessary for plant growth and development. They have different
function in the vegetal organism. These functions can be grouped in:
• specific functions, in case the mineral ion has a certain role and cannot be
substituted by another element (a catalyzing and structural role, a role in
ensuring the colloidal status of the protoplasm, in electron transport during a
specific process etc.).
Mineral ions can act like biocatalyzers, in which case they:
• are part of the prosthetic groups of proteins (cytochrome enzymes, from the
electron transport chain);
• are part of the catalytic site of the enzymes (Cu in peroxidase, Fe in catalase)
• are enzyme activators (K does not enter in the composition of any organic
substance, but it activates about 50 enzymes by modifying the conformational
structure of the proteins).
Chemical elements (like Mg, Zn etc.) merge with enzymes and form compounds
called chelates, which enable enzyme interaction with their substrates. They can
also be found in the composition of specific amino acids (S), proteins (N, S),
nucleic acids and ATP (P), chlorophyll (Mg), cellular wall (Ca) etc., where they
have a structural function.
The function of protoplasm colloidal status regulation is carried by K ions which
act to decrease the viscosity of the protoplasm and by Ca ions that act to increase it
thus influencing the hydration level of protoplasmic colloids;
• nonspecific functions—when performing these functions an ion can be
substituted by another one (osmotic regulation—inflow and elimination of water
from the cell is determined by the quantity of ions and not by their type).
The physiological significance of mineral elements has been studied extensively
(Table 6.1). It is known that mineral elements influence the exchange of substances,
alter cell turgidity and the permeability of biological membranes, enter in the
6.1 Importance of Mineral Elements in Plant Nutrition 153
Plants, depending on their age, species and organ contain 90–98 % water, the rest
being represented by dry mass which contains the following average amounts of
elements: C—45 %, O—42 %, H—6.5 %, N—1.5 %. At high temperatures,
organic substances are burned and eliminated, while the remaining mineral ele-
ments form the ash, which makes up 0.2–20 % of the dry mass. The total amount of
ash varies depending on the species, on the nature and age of the organs and is
tightly connected to the composition of the soil and its humidity.
The chemical composition of the plants reflects their need in mineral elements.
Burning different organs and plant parts result in different amounts of ash. Wood,
for instance, contains—1 %; seeds—3 %; leaves—5–15 %; bark—7 % of the dry
weight. These figures demonstrate that mineral elements are concentrated in cells
and organs with a high vital activity. All the elements of the Mendeleev table are
154 6 Mineral Nutrition of Plants
part of the ash composition. This allowed the Russian scholar V.I. Vernadski to
mention the role of plants in the circuit of elements in nature (especially of the rare
ones).
The content of mineral elements in ash varies within certain limits. Depending of
the percentage of elements in the ash and in living plants, they can be classified as
follows:
• macroelements (O, C, H, S, P, Na, Ca, K, Mg), which represent at least 0.01 %
of the plant’s dry weight;
• microelements (Fe, Co, Cu, B, I, Mn, Mo) which make from 0.01 to 0.001 % of
the dry mass of the plants;
• ultra microelements (minor traces of elements).
The root is the organ which grows into the soil having the function to fixate the
plant and to absorb water and mineral salts (see: Chap. 3, The water regime.). Roots
can form mycorrhizae (symbiotic relationships between the root and some micro-
scopic fungi), bacteriorrhizae, (symbiosis between the roots and bacteria).
If the tip of a young root is analyzed with a magnifier, five regions can be
distinguished (Fig. 6.2): the root cap (calyptra), the apical meristem (division zone),
the elongation region (smooth), the root hair region (differentiation zone) and the
mature region (rough).
The root cap region protects the tip of the root and is also called the calyptra.
This region has a constant length, because new cells are constantly born to replace
old ones that disappear as a result of friction with the soil.
The apical meristem is the region where cell division promoting root growth
occurs. It gives birth to the primordial meristem.
The elongation region is a root region where cell do not divide but rather
increase their longitudinal dimensions manyfold.
The root hair region is a region where cell differentiation occurs. Here, histo-
genesis gives birth to the rhizoderm, to the epidermis and to the central cylinder of
the root. Rhizoderm cells differentiate and form cylindrical projections perpendic-
ular to the root axis, hence the name of this root zone.
The function of the root hairs is to absorb, through endosmosis, water and
mineral elements from the soil. They have a thin cellulosic membrane, a nucleus
located close to the tip and surrounded by cytoplasm, while the center of the cell is
filled by a big vacuole. Root hairs help create huge absorption surfaces that can
reach tens or even hundreds of square meters.
The maturation region has a rough dentate surface due the root hairs that have
detached in the process of constant renewal.
156 6 Mineral Nutrition of Plants
Fig. 6.2 Root structure. The main root regions and tissue types are represented
The macroelements are the mineral elements which constitute at least 0.01 % of the
plant dry weight. They are part of the tissue composition and have a key-role in
plant growth and development.
Nitrogen (N) was discovered by D. Rutherford in 1772 in the form of a gas, which
cannot maintain the processes of burning or life. It is, however, part of the most
6.5 Physiological Role of Macroelements 157
To reduce the nitrates the following are necessary: metabolic energy, the pres-
ence of proton and electron donors which are NADPH+H+ or NADH+H+ (reduced
nicotinamide adenine dinucleotides). The source of these substances in the root
system is represented by the process of respiration, while in leaves—by photo-
synthesis (nitric photosynthesis).
Thus, nitrate reduction is mainly determined by the intensity of these processes.
For proper respiration, a certain quantity of carbohydrates (respiratory substrate) is
sufficient. If the quantity of carbohydrates is reduced artificially below this level,
nitrites will not be reduced, and will accumulate instead in all the organs of the
plant. An excessive accumulation of nitrites in plants may have a negative (toxic)
impact on them.
Nitrate recovery is also stimulated significantly by light. Presumably, for these
reactions the products resulting from acyclic photophosphorylation (NADPH+H+,
ATP) can be used directly. Blue light is stimulatory in this process due to the fact
that flavin, which is an integral part of nitrate reductase, absorbs blue light and is
activated by it.
Another nitrogen source for plants is the ammonium cations, which enter plant
tissues very easily (faster than nitrites) and are used directly.
Amination reactions and amino acid biosynthesis. The ammonium cation is a
basic component of plant metabolism. Its origin can be:
• directly from the soil;
• formed as a result of nitrite reduction;
• formed as a result of protein catabolism in aging organs.
Accumulation of ammonium in cells may have a negative impact. However,
plants can sequestrate ammonium with the help of organic acids forming amides
(glutamine, asparagine etc.). This process is similar to that of forming urea in animal
organisms. When the intensity of respiration is low, or in case of carbohydrate
insufficiency, amides are not formed and NH4+ accumulates, which causes plant
intoxication.
There is a whole group of plants which accumulate a big amount of organic
acids that are used to neutralize ammonium. This caused the division of plants into:
(a) those that form amides (for instance asparagine and glutamine) and (b) those that
form ammonium salts.
6.5 Physiological Role of Macroelements 159
A change in the pH of the cytoplasm, may alter the nitrogen metabolism causing
a switch between amide and ammonium salt formation.
Amino acid and amide biosynthesis pathways in plants. During respiration,
the α-ketoglutaric and oxalic organic acids are produced as intermediary products.
These acids incorporate ammonia as a result of a direct amination reaction:
The quantity of formed asparagine and glutamine and their importance differ
depending on the plant species and environmental conditions. Glutamine is formed
in leaf and root cells. Formation of asparagine predominates during protein
catabolism in seeds. Thus, asparagine is the neutralization form of the NH3 formed
during protein degradation (the regressive branch of nitrogen exchange in plants):
The age of the plant plays an important role in amide formation. The younger the
plants, the higher its ability to form amides. In younger organs and even in younger
cells of the same organ, the intensity of amide formation is higher.
Crude sap and guttation sap both contain amides which proves that the nitrogen
absorbed from the soil is transformed into amides in the living cells of the root.
Amides in plants represent:
• the form of NH3 neutralization;
• the form of transport for nitrogen compounds;
• a building block for the synthesis of other amino acids in the process of
transamination.
Transamination and protein synthesis. Each of the amino acids which are formed
by direct amination (the glutamic and aspartic acids) are predecessors for a whole group
of amino acids. Out of the 20 aminoacids which enter in the composition of plant
proteins (proteinogenic amino acids), only two can be formed in the process of direct
amination. The others are formed as a result of transamination and retransformations.
These reactions are catalyzed by special enzymes—aminotransferases and occur with
6.5 Physiological Role of Macroelements 161
participation of the pyridoxal phosphate coenzyme (the active form of the B6 vitamin).
The role of this catalyzer consists in ligating the amino group and forming pyridoxamine
phosphate and keto acids. The most common reaction is that by which the amino group
is dissociated from the glutamic acid. Different amino acids synthesized by transami-
nation produce other amino acids by transforming the carbon backbone. Thus amides
serve as donors of amino groups.
Plants, in comparison to animals, have the ability to synthesize all the necessary
amino acids. They can be formed in different organs of the plant—leaves, roots, the
apex of the stem:
Transamination
Some amino acids are formed directly in the chloroplasts, where they are used to
synthesize proteins.
Protein synthesis occurs is more intense in meristematic tissues undergoing
development. Protein biosynthesis stops in cut leaves, which proves the require-
ment of factors formed in plant roots (mot probably a phytohormone from the group
of cytokines or a similar substance):
Deamination
(3) a high intensity of respiration and phosphorylation (ATP is necessary for all
the stages of nitrogen compound transformation: recovery of nitrates, forma-
tion of amides, activation of amino acids in the process of protein synthesis);
(4) presence of the basic components of the proteosynthetic apparatus:
• DNA—as the molecule encoding the information about the sequence of
amino acids in the protein chain,
• mRNA—the agent transmitting information from DNA to ribosomes,
• tRNA—associates the nucleotide triplets (codons) of the mRNA with their
corresponding amino acids decoding the nucleotide sequence into the
amino acid sequence;
(5) presence of the ribosomes—the structural units of the cell which carry protein
synthesis;
(6) presence of enzymes catalyzing protein biosynthesis (aminoacyl-tRNA-syn-
thetases etc.) and protein factors;
(7) Mineral elements (Mg2+, Ca2+).
The progressive branch of nitrogen exchange in plants, found mainly in young
organs, ends up with protein synthesis (primary synthesis of proteins). However, a
continuous process of protein decay also occurs in plants in parallel. Protein
renewal takes place rather rapidly (about 60 % of the proteins are renewed during
48 h).
Proteins are degraded to amino acids and further to NH3 (the regressive branch
of nitrogen exchange), which is later again neutralized by the formation of aspar-
agine and glutamine that serve for amino acid synthesis. This process allows plants
to form a new set of amino acids, which are used in protein anabolism (secondary
synthesis of proteins).
Thus, both mineral and organic nitrogen can be found in vegetal organs. Organic
nitrogen is represented by micromolecules (amides, organic acids, nitrogenous
bases etc.) and by macromolecules (proteins, nucleic acids). 80–85 % of all the
nitrogen in the vegetal mass is represented by enzymes, while in seeds, by reserve
substances such as proteins.
The symptoms of nitrogen deficiency in plants. Nitrogen deficiency causes
delay in plant growth and development, plants turn green-yellow. The lower leaves
turn yellow (chlorophyll molecules are destroyed and nitrogen compounds are
transported to younger leaves), the vegetation period is reduced, the intensity of
protein and enzyme biosynthesis is diminished, which has an impact on cellular
metabolism. In higher plants anthocyanin biosynthesis intensifies.
Atmospheric nitrogen fixation. Some plant species (leguminous plants), which
live in symbiosis with microorganisms of the Rhizobium genus have the capacity to
fix atmospheric nitrogen. About 500 species of microorganisms belonging to this
genus are known, which coexist with higher plants by forming specific root nodules
and recruiting free nitrogen from the atmosphere (Figs. 6.4 and 6.5). These
microorganisms contribute to atmospheric nitrogen conversion into ammonium
6.5 Physiological Role of Macroelements 163
Fig. 6.4 Root nodules containing nitrogen fixing bacteria of the genus Rhizobium
ions by using the necessary energy and metabolites from higher plants (Figs. 6.6
and 6.7).
The interaction between bacteria and the plant is triggered by certain flavonoids
synthesized by the roots, which induce the expression of genes in microorganisms
that are involved in nodule formation.
164 6 Mineral Nutrition of Plants
Nitrogen fixing bacteria have the enzyme nitrogenase encoded by genes of the fix
and nif type. This enzyme consists of two proteins which contain Mo and Fe and
one which contains Fe. The genes are active in anaerobic conditions, while the level
of oxygen in nodules is regulated by leghemoglobin, the protein part of which is
encoded by the plant while the heme—by the bacteria.
6.5 Physiological Role of Macroelements 165
Sulfur belongs to the category of nutritive elements which are absolutely necessary
for vital activity. This element enters the plant in the form of sulfate ions (SO42−).
Its amount in plants is relatively low, making up 0.2–1.9 % of the dry mass.
The importance of sulfur. This element is part of cysteine and methionine
which are among the most important amino acids given their role in catalytic
reactions mediated by enzymes. These amino acids can be found in a free state or as
residues in protein sequences.
Methionine is one of the essential amino acids and has some unique properties
due to the sulfur and the methyl groups. It was identified in the active centers of
many enzymes. Methionine confers hydrophobic properties to protein molecules,
which plays an important role in stabilizing their structural conformation. One of
the most important functions of sulfur in proteins and polypeptides is the partici-
pation of SH groups in the formation of covalent, hydrogen and mercaptide bonds
which determine the three-dimensional structure of proteins. Disulfide bridges—
covalent bonds between the thiol groups of, for instance, two cysteine residues
166 6 Mineral Nutrition of Plants
H2 SSOSO3 2 SO4 2
forms in plant organs depends on the intensity of sulphate reduction and assimi-
lation, on the concentration of SO42− in the nutritive environment.
A part of the sulfur which was absorbed by plants is retained in the roots. The
biggest part of it, however, moves from the roots into the xylem vessels, from
where it is transported to the fast growing young organs, which are involved in a
dynamic metabolic process.
From the leaves, sulfate and reduced sulfur forms (amino acids which contain
sulfur and glutathione) enter the phloem and are deposited. In seeds, sulfur is
present in the organic form, but when growing, it is partially converted to the
oxidized form. At the same time, during seeds maturation, reduction of sulfur and
synthesis of sulfur-containing amino acids takes place.
A summary of the process of complete sulfur reduction can be expressed by the
following formula:
CysteineHomocysteineMethionine
Phosphorus (P) is part of the essential nonmetal group and forms 0.2 % of the total
dry mass. This element enters the plant in its oxidezed PO43− form. After under-
going transformations it preserves its state and degree of oxidation.
The importance of phosphorus. In plant tissues, phosphorus is present in
organic (proteins, nucleotides, vitamins and other compounds) and inorganic form
(orthophosphoric acid and its salts).
The phosphate group determines the hydrophilicity of a phospholipid molecule
while its lipid part remains hydrophobic. This is why, at the border of phase
separation in biological membranes, the phospholipids are oriented with their
phosphate ends outside, while lipophilic groups are retained stably inside the lipid
bilayer, stabilizing the membrane.
Another unique function of phosphorus is its participation in the process of
phosphorylation of cellular proteins mediated by protein kinases. This mechanism
is a means of regulating many metabolic processes, because inclusion of the
phosphate in the protein molecule causes a redistribution of electric charges in it
and, as a consequence, leads to changes in its conformation and, consequently, its
function. Protein phosphorylation regulates such processes like ARN and protein
synthesis, is important in signal transduction and many other processes etc.
Phosphorus is the basic bioenergetic element which has an important limiting
role in plant growth and development. Its action is also crucial for entering and
exiting states of decreased metabolism (e.g. anabiosis).
The symptoms of phosphorus deficiency. The deficiency of phosphorus has an
impact on all vital processes—photosynthesis, respiration, plant growth and
development. It produces a change in leaf color into blue-green with purple and
golden shades, which is a result of the delay in protein synthesis and carbohydrate
accumulation. Leaves become smaller and thinner. Plant growth as well as fruit
ripening (maturation) is delayed. During phosphorus scarcity O2 absorption speed is
reduced and the activity of enzymes which participate in respiration is altered.
Plants are more sensitive to phosphorus deficiency during the primary stages of
their growth and development. A proper phosphorus input, at later stages, causes
fast maturation of plants and fruits.
Forms of phosphorus in nature. Phosphorus reserves in soil are relatively
small (2.3–4.4 t/ha—P2O5). Out of this quantity two thirds are represented by the
mineral salts of the ortophosphoric acid (H3PO4) and one third—by organic
compounds (from dead remnants, humus). The concentration of phosphorus in the
soil solution is low (0.1–1 mg/l). Organic compound containing phosphorus are by
large insoluble in this solution. But some agricultural species (buckwheat, peas) use
poorly soluble forms as well. Phosphorus from organic remnants and from humus is
mineralized by microorganisms and the biggest part of it is converted in poorly
soluble salts which can be assimilated by plants and converted to mobile forms.
This is possible due to the fact that roots release organic acids that acidify the
rhizosphere contributing to the sequence of transformations:
6.5 Physiological Role of Macroelements 169
Phosphorus is part of a big list of organic compounds such as: nucleic acids (DNA
and ARN), nucleotides, phospholipids, vitamins, and plays an important role in the
exchange of substances in plants.
Many vitamins which contain phosphorus and its derivatives are coenzymes and
are directly involved in cell catabolism, accelerating the substance exchange pro-
cesses. Phosphorus has the specific ability to form instable, macroergic bonds, this
representing a convenient, controllable, recyclable means of energy utilization in
different biochemical and physiological processes.
When phosphoric acid reaches the root cells, it is rapidly included in the com-
position of nucleotides forming AMP (adenosine monophosphate) and ADP
(adenosine diphosphate). Later, ATP is formed as a result of substrate and oxidative
phosphorylation (the aerobic and anaerobic phases of respiration). After just 30 s
from the moment of its absorption by the plant, phosphorus is included ATP
molecules, which is later used to activate amino acids, in the process of nucleic
acids synthesis etc.
Potassium or kalium (K) is one of the most important and necessary nutritive
elements. Its amount in plants varies between 0.5–1.2 % of the dry mass. The
content of potassium in the cell is hundreds of times higher than in the environment,
thus for quite a long time the ash was the only source to obtain this element.
The reserves of kalium in soil are significant and can be found in the following
forms:
• in colloidal particles;
• in the composition of organic remnants and microorganisms;
• as mineral salts dissolved in the soil solution.
The most accessible source of potassium are the 0.5–2.0 % of the soil reserves.
Periodic drying and wetting of the soil, the activity of the root system favors
potassium conversion into accessible forms. Fertilizers containing potassium are
water soluble and, by entering the soil, they interact with its colloids from where it
can be absorbed by plants. However potassium input displaces other ions (H+, Ca2+,
Al3+, Mg2+) from the colloids into the soil solution. Kalium fertilizers are physi-
ologically acid salts which stimulate HCI and H2SO4 accumulation in the soil
That’s why, in acid soils the efficiency of potassium fertilizers decreases.
In plants, higher amounts of potassium can be found in young, growing tissues,
which are characterized by high rates of substance exchange—meristems, cam-
bium, young leaves, buds, sprouts.
In cells, potassium can be found in the ionic form. It is not part of organic
substances, it has a high mobility and is easily recyclable. The rapid migration of
kalium from mature cells to young ones happens due to sodium, which replaces it in
cells that stopped growing.
6.5 Physiological Role of Macroelements 171
Fig. 6.11 Role of potassium and chlorine in stomatal opening and closure
172 6 Mineral Nutrition of Plants
• Potassium is absorbed by plants as cations and only forms weak bonds with
different compounds of the cell. This is one of the reasons why it creates an ionic
asymmetry and an electrochemical potential at the membrane separating the cell
and the environment.
• It is a cation which activates fermentation systems. Nowadays, more than 60
enzymes activated by potassium are known. It is needed for including phosphate
in organic compounds, in the reactions of phosphate group transfer, for protein
and polysaccharide synthesis. It participates in the biosynthesis of riboflavin—a
component of all flavin dehydrogenases.
• Potassium increases the accumulation of starch in potatoes, of sucrose in sugar
beet, of monosaccharides in fruits and vegetables, of cellulose, hemicellulose
and pectic substances in the cell wall. As a consequence, it increases the strength
of the stems in cereals, the quality of hemp and flax fibers is improving.
A sufficient supply of kalium in plants increases their resistance to diseases
caused by fungi and bacteria. If the levels drop, the quantity of Na, Mg, Ca NH3,
and of the H+ ions increases to compensate the osmotic effect, but they cannot
compensate for its specific functions.
The critical period for supplying plants with potassium is 1–2 weeks after their
appearance. However, the biggest quantity is absorbed during the period of vegetal
mass growth.
In case of scarcity, leaves start turning yellow from plant bottom to its top (from
old to young leaves). Leaves start changing color from the edges, later the edges
and the tips turn brown with red spots (rust color), then their death and decay
occurs. The scarcity is strongly felt in young and in actively growing organs, this is
why the functionality of cambium is reduced, the development of conducting tis-
sues is disturbed, the epidermis and the cuticle become thinner, the processes of cell
multiplication and elongation are delayed. Shortening of the distance between in-
ternodes leads to dwarf phenotypes. Potassium deficit causes a drop in the domi-
nating effect of the apical bud, lateral sprouts develop more intensely and plants get
a shrub-like aspect. This deficit also causes the productivity of photosynthesis to
drop. In this case, neither phosphorous nor nitrogen containing fertilizers can
replace potassium.
Calcium (Ca) is another chemical element which is important for plants
metabolism. The general quantity of Ca2+ in different plant species is of 5–30 mg/g
of the dry mass. Depending on their “attitude” towards Ca2+, plants can be divided
into three groups:
• calciphiles;
• calciphobes;
• neutral.
Legumes, buckwheat, sunflower, potatoes, cabbage and hemp are reach in cal-
cium ions. Grasses, flax and sugar beet are much poorer in calcium.
The tissues of dicotyledonous plants usually contain much more calcium than
those of the monocotyledonous ones. Ca2+ is accumulated and stored in older
6.5 Physiological Role of Macroelements 173
organs and tissues. This is due to the fact that transportation happens through the
xylem which makes its reutilization difficult. When cells grow old or when their
physiological activity drops down, Ca2+ moves from the cytoplasm into the vac-
uoles and is stored in the form of insoluble salts of the citric and oxalic acids etc.
These crystals impede the reutilization of this cation. In most crop plants, Ca2+
accumulates in vegetative organs. In the root system Ca2+ can be found in the form
of phytin and its quantity is lower than in aerial organs.
In cells, a big quantity of Ca2+ is bound by pectic substances from the cell wall.
It is also contained in chloroplasts, mitochondria and nucleus, complexed with
biopolymers.
Ca2+ performs different functions in the process of exchange of substances.
These functions depend on calcium influence on:
• the structure of membranes, determining ion circulation through them and the
bioelectrical processes;
• the processes of cytoskeleton transformation—actiniform proteins, which par-
ticipate in the processes of cytoplasmic flow, in the reversible processes during
which its viscosity is altered (conversion from sol into gel and vice versa), in the
spatial organization of the enzymatic cytoplasmic systems (e.g. glycolysis).
Calcium is necessary for processes of plants secretion. This element activates a
series of fermentative systems in the cell—dehydrogenases, α-amilases, lipases,
phosphatases etc. in this case, Ca2+ can determine the assembly of protein subunits,
can serve as a bridge between the enzymes and the substrate, can modulate enzy-
matic action allosterically. Ca2+ surplus in ionic form depresses photophosphory-
lation and oxidative phosphorylation.
The regulatory action of Ca2+ on metabolism depends on the interaction with the
intracellular receptor of calcium—calmodulin protein (recruits 4 ions of Ca2+). The
Ca2+—calmodulin complex activates several enzymatic systems.
The ions of Ca2+ have the important role to stabilize membranes. By interacting
with the negatively charged phospholipids groups they stabilize the membrane and
decrease its passive permeability. In case of a deficit, membrane permeability
increases, fragmentation and ruptures appear and the process of membranous
transport is disturbed.
By limiting the absorption of other ions into the organism, Ca2+ counters the
toxicity determined by the surplus of ammonium ions, Fe, Al and Mn, increases
plant tolerance to salts, decreases soil acidity. It is Ca2+ which performs the role of
ionic balancer in creating equilibrated physiological solutions, since its quantity in
soil is high.
In case of calcium scarcity the first to suffer are young meristematic tissues and
the root system. In multiplying cells, new cell envelopes are not formed and, as a
consequence, multinucleate cells are produced. Formation of lateral roots and root
hairs is ceased, root growth ceases. Ca2+ insufficiency causes pectic substances to
swell which leads to the appearance of mucus on cell envelopes and cell destruc-
tion. As a consequence, the roots, leaves, some parts of the stem rot and die. The
tips and the edges of the leaves first turn white and later, black, the leaf blade loses
174 6 Mineral Nutrition of Plants
its shape and twists. On the fruits, in the conducting and reserve tissues necrotic
regions appear. The structure of the plasmalemma and of the cellular organelle
membranes is distorted.
Most types of soil are rich in calcium and scarcity is very infrequent (in soils
with increased acidity and in those with high salt content).
Magnesium (Mg), in terms of its quantity in plants, occupies the fourth position
after K, Na and Ca. In higher plants, its content in the dry mass is as high as
0.02–3.1 %. It is usually found in the so called short day plants—maize, millet,
sorghum, potato, beet, and tobacco.
A kilogram of fresh leaves contains 300–800 mg of magnesium. Of them,
30–80 mg are part of chlorophyll. The biggest part of magnesium is contained in
young leaves and in the tissues in which reserve substances are stored. In seeds,
magnesium is accumulated in embryos, where its levels are several times higher
than in the endosperm. About 10–12 % of the magnesium which is part of the
chlorophyll performs a unique function in the vegetal body. Magnesium is needed
for the synthesis of protoporphyrin IX—the direct predecessor of chlorophyll.
During daylight, Mg2+ ions are released from the thylakoids into the chloroplast
stroma. When Mg2+ concentration in the stroma goes up, the RDF-carboxilase and
other enzymes are activated. It is supposed that the increase of Mg2+ concentration
(up to 5 mol/l) causes the activation of CO2 reduction.
Magnesium influences directly the enzymatic conformation the enzyme and the
proper conditions for its functioning, determining the pH of the cytoplasm as an
anti-ion of the protons. Magnesium influences a series of reactions of electron
transport during phosphorylation:
(a) NADP+ recovery;
(b) the speed of the Hill reaction;
(c) Electron transfer from PS I to PSS II.
In most of the cases, the influence of Mg2+ on other processes of substance
exchange is connected to the ability of enzyme regulation, so its importance for a
series of enzymes is unique. Mg2+ is the coenzyme of all the enzymes which
catalyze phosphate groups transport (phosphokinases, phosphotransferases, ATP-
ases, pyrophosphatases). This happens due to the ability of Mg2+ to form com-
plexes. Mg2+ is also needed for many enzymes involved in glycolysis and for those
of the Krebs cycle. In case of its scarcity, the number of mictochondrial cristae is
reduced, their form is altered and consequently they disappear. In 9 out of the 12
reactions of glycolysis, participation of activating metals is necessary and magne-
sium participates in 6 of them. Mg2+ is also necessary for the activity of enzymes
which participate in lactic and alcoholic fermentation; it accelerates the synthesis of
ethereal oils, calcium, and A and C vitamins; it is necessary for the formation of
ribosomes and polisomes, for the activation of amino acids and protein synthesis
and is used in all the processes in a quantity of at least 0.5 mol/l. Magnesium
activates ADN and ARN-polimerases.
The process of Mg2+ absorption by the plant, depends on its supply with other
cations. Thus, if a big quantity of K+ or NH4+ is contained in the soil, the level of
6.5 Physiological Role of Macroelements 175
magnesium is decreasing, especially in vegetative organs. Ca2+ and Mn2+ also act
like competitors in the process of Mg2+ absorption by plants. The Ca/Mg ratio has a
great importance in the vital activity of the plant and regulates many metabolic
processes.
When the soil pH value is decreasing, Mg2+ enters plants in lower amounts. The
insufficiency of magnesium in plants is felt at the level of 2 mg per 100 g of soil.
Mg2+ deficit manifests by the formation of yellow-green spots in the vicinity of the
veins. The edges of the leaf blade become yellow, red, orange (Fig. 6.12).
flax, the hop, the ricin, the maize and the grapevine. The symptoms of scarcity differ
from species to species and are normally exhibited by a reduction in plant growth:
short internodes in grasses, a “rosette” arrangement of the terminal leaves and
brunches, emergence of yellow spots on the leaves. In apples, apricots, plums,
cherries and grapevine zinc insufficiency causes the formation of small leaves with
chlorotic spots.
Absorption (through the differentiation region of the root) and transportation (radial
and xylemic transport) of the ions is carried out by the same pathways as water but
at a lower speed. In the process of mineral element absorption two phases have been
delimited:
• the rapid phase of absorption and desorption;
• the slow phase of absorption and desorption.
The rapid phase is linked to the absorption and desorption of the ions at the level
of the cell wall, while the slow phase—at the level of plasmalemma.
Absorption takes place through the entire root system (the total volume of
absorption), but in an active manner it occurs only at the level of the root hair region
(the active volume). The first barrier in the way of mineral elements entrance into
6.7 Mechanism of Absorption and Transport of Ions in Plants 179
the cells is the cell wall. Since it has a fibrillar structure, the ions cross the cell wall
through:
(1) diffusion;
(2) exchange of ions;
(3) adsorption.
Diffusion is a physical phenomenon through which the atoms or molecules of a
substance slowly mix with the molecules of the other substances which they con-
tact. At the cell wall level it happens through the free spaces of the macro- and
microfibrils.
Absorption is obtained through mutual ion exchange between the cells of the
plant and the soil solution, in which the cell wall has the role of a cation exchanger.
This is possible due to the carboxylic groups (-COOH) of the pectic acids, which
participate in the exchange of hydrogen ions with monovalent cations of potassium
(K+), sodium (Na+), ammonium (NH4+) as well as due to proteins, which contain
both carboxylic (-COOH) and amine (-NH2) groups. These functional groups have
the property to accumulate cations and anions from the soil solution.
The exchange of ions also happens due to the respiration process. The carbon
dioxide which is released as a result of this process, being very soluble in water,
forms carbonic acid, which dissociates to generates anions of HCO3−, CO3− and
cations of H+ able to participate in ion exchange with the soil solution. Thus, in the
cell wall and the plasmalemma one can find ions of H+, HCO3− and OH−, which are
released by the roots back into the soil, by absorbing in turn ions of NO3−, PO43−,
K+, Ca2+, Mg2+, etc.
Another mechanism of ion uptake by the cell wall is the adsorption phenome-
non. There are two types of adsorption: mechanical and by means of chelates.
The mechanical adsorption is performed via very labile, transient bonds between
the adsorbed substance and the cell wall. Adsorption by means of chelates is
stronger and is carried out with the formation of stronger bonds between mineral
ions (cations or anions) and organic substances.
Ion penetration through biological membranes, which is the next obstacle,
can be either active or passive.
Passive transport through biological membranes is carried out according to the
concentration gradient, without metabolic energy consumption and the process is
connected with diffusion. From the thermodynamic point of view, the direction of
the diffusion process is determined by the chemical potential of the substance. The
higher the concentration of the substance, the bigger the chemical potential will be.
Diffusion is oriented towards a lower chemical potential. Importantly, the direction
of ion movement is also determined by the electrostatic potential. In ion diffusion,
the values of the kinetic molecular energy count. This energy grows with tem-
perature increase and concentration increase. Ions with different charges have dif-
ferent diffusion rates through the semipermeable membranes. This fact leads to a
difference in the electric potential, which can serve as a driving force for trans-
ferring other ions. The electric potential can also appear as a result of irregular
180 6 Mineral Nutrition of Plants
Fig. 6.13 The activity of Mg2+, Na+, K+—ATPase in the calathides and leaves of different
sunflower genotypes in the flowering phase (μmol ATP/min g fresh weight) (Glijin 2002)
ions have to pass yet another barrier—the tonoplast. The permeability of the
tonoplast is lower than that of the plasmalemma. It is known that there are two
active systems of ion transport through the cell:
• located in the plasmalemma and which functions in conditions of low intra-
cellular concentrations;
• located in the tonoplast and functioning only at high intracellular concentrations,
when the cytoplasm, is saturated with these ions.
It is the second mechanism which is responsible for ion entry into the vacuoles.
Thus, ions which enter through the plasmalemma are used for cellular needs or are
transported into the neighboring cells and only their surplus enters the vacuoles.
Transportation of mineral salts in plants are carried out by means of 2 paths (radial
and xylem). The mineral substances which are accumulated in the cytoplasm of root
hair cells and in cortical cells are transported towards the xylem vessels (radial
transport) through the symplast and apoplast (Fig. 6.14). Transport of ions through
the apoplast is based on the process of diffusion of ions at the cell wall level. The
symplastic path is permanently active and occurs from cell to cell through the
plasmodesmata. Ions from the soil solution are transported through the apoplast to
the plasmalemma simultaneously in all cortical cells. The ion pumps from the
plasmalemma of root hair cells and cortical cells function in the same direction,
transporting ions from the apoplast into the symplast and vacuoles, where they
generate the turgor pressure, or from cell to cell towards the conducting vessels.
182 6 Mineral Nutrition of Plants
Transport of the crude sap from the roots towards the leaves is performed
through the xylem. The crude sap is a diluted solution which contains ions and
organic compounds which originate from the soil. The delivery of mineral sub-
stances and water to the leaves is carried out through their branching veins (xylem).
The xylem transport of minerals follows the water stream up to the level of the
foliar system, but at a lower speed.
Movement of the nutritive substances in an ascendant manner through the xylem
is a passive process. But the distribution of nutritive substances is not determined
by the intensity of respiration, but by the exchange of substances in the respective
organ and by the presence of auxins in the growing apex which has the role of a
dominant center. Its removal leads to a uniform distribution of mineral ions in all
plant organs.
The distribution of ions is determined by the functional activity of the tissue. The
biggest quantities of ions are provided to the young growing tissues.
The soil represents the main substrate for mineral nutrition and is formed by three
types of substances (solid, liquid and gaseous). It is characterized by a certain
chemical composition, pH, structure etc.
The soil solution is a physiologically equilibrated solution, in which ion
antagonism is present—reducing the negative effects of certain ions by others. The
concentration of the soil solution is 0.05–0.15 %.
Depending on the pH of the soil, selective absorption of certain ions occurs.
Anions are absorbed at a weak acidic pH, while cations—at a weak basic pH. At the
6.8 Soil as a Substrate for Plant Nutrition 183
optimal pH the monovalent anions and cations are absorbed at the same rates. The
salts that do not affect the soil pH are called neutral:
The speed of cation absorption is higher than that of anion absorption. If the soil
contains many anions it is called physiologically acid soil:
Glossary
References
Rubina AB (ed) (1980) Results of science and techniques. Plant physiology T. 4. Ion
Transportation in plants, p. 176
Şcolinik MIa (1974) Microelements in the life of a plant L. p 324
Şeveakova NI (1979) The metabolism sulfur in plants M. p 166
Sîtnic KM, Kniga NM, Musatenko LM (1972) Root physiology Kiev. p 356
Chapter 7
Plant Growth and Development
Historical Background
Brief Updates
Parasitic bacteria Agrobacterium tumefaciens and A. rhisogenes use phytohormones
to induce tumors in plants. A. tumefaciens contains the Ti plasmid, harboring the
iaaM1 and iaaH2 genes involved in the synthesis of auxin (IAA) while the ipt gene
encodes cytokinin biosynthesis. Because these genes lead to uncontrolled cell
proliferation, they were assigned to oncogenic agents.
The Arabidopsis thaliana gene Leafy (LFY) is involved in plant transition from
the vegetative to the reproductive state, contributing to the initiation of flower
formation. In addition to this gene the Apetala (AP1) and cauliflower (CAL) genes
are involved in flower formation. An opposite effect is exerted by the terminal
flower (TFL1) gene, which retains the formation of flowers, inhibiting gene
expression of LFY and AP1. Genes with similar effect were detected in rice
(OsRCN1) and rye (LpTFL).
Jasmonic acid (JA), which is synthesized from linolenic acid, by the octadecanoic
pathway has been found in higher plants and is one of the key phytohormones
involved in stress signaling. This phytohormone manifests an inhibitory effect on
photosynthesis, callus, growth, cell division and DNA replication and induces
senescence. Exogenous treatment of the Quercus ilex plant with jasmonate decreased
the rate of photosynthesis, induced an alteration in the stomatal conductance levels
and determined the elimination of volatile monoterpenes and methyl salicylate.
Vegetable bodies absorb water and mineral salts, accumulate solar energy, make
countless reactions of substance exchange. As a result of these activities plants
grow and develop. Growth and development are two specific integrated processes
of any living organism occurring simultaneously. Growth and development occur
throughout ontogeny (from gr. ontos “being” and genesis, “origin”) from zygote
until death. During ontogeny the hereditary information (genotype) is implemented
in strict relationship with the conditions of the environment which ultimately results
in a phenotype characteristic for the species.
Growth represents a combination of physiological and biochemical processes
through which the irreversible increase in volume, mass, size of the plants happen
due to tissue and organ expansion (Fig. 7.1).
Unlike animals, plant growth can occur throughout the life, because the centers
of growth (stem and root tips, shoots, cambium, phellogen) have active meriste-
matic tissues that work continuously. The process of growth in plants is rhythmi-
cally interrupted by periods of rest (latency). Latency in plants is a normal
periodical phenomenon which is genetically programmed and during which a
decrease in the intensity of life processes occurs.
Growth intensity varies depending on the species, organ, environmental condi-
tions etc. The highest rates of growth were found in mushrooms (5 mm/min) and in
190 7 Plant Growth and Development
Fig. 7.3 Tissue differentiation in different organs of the plant (Gilbert 2000)
Fig. 7.4 The transition from vegetative to reproductive development (Gilbert 2000)
Plants or organs that are in biological repose are characterized by the accumu-
lation of growth inhibitors and by certain physiological and biochemical processes
that prepare future growth. This is part of the reason why growth is not restored
even when all the necessary conditions exist.
Usually, plants enter the forced dormant state in the absence of one of the factors
necessary for growth and as soon as these factors become available, growth pro-
cesses are resumed. Plants exit forced dormancy only when the biological repose is
due to begin. Perennials fall in deep dormancy in autumn and in late winter they
long before budding they enter forced dormancy.
Either the whole organism or pats of it (seeds, tubers, roots) can be dormant.
Under certain conditions some plant organs can grow while others (buds) are
dormant. Transition to the repose state is often accompanied by loss of organs
(falling leaves or even entire shoots). It is in such a state that perennials survive
winter.
While dormancy can be different and can affect different organs differently there
are also common features that characterize the phenomenon:
(1) lack of active growth (there may be a latent growth);
(2) decrease in the intensity of metabolic processes;
(3) reduction in the amount of growth promoters.
In most of the crop plants the state of repose is controlled by photoperiodism.
Long days accelerate vegetative growth and short days lead to growth inhibition
and formation of dormant buds.
There are different ways to stop this phenomenon (etherization, hot baths,
treatment with volatile substances or low temperatures, growth stimulating sub-
stances) which are used in greenhouses for growing winter flowers. Bud and other
organ dormancy may be interrupted with gibberellins, cytokinines, ethylene and
triggered by abscisic acid.
7.2 Types of Plant Growth 193
The basis for multicellular organism growth consists in increasing the number and
size of the cells, accompanied by differentiation. Growth and development start
from a single cell and follow three phases:
Embryonic stage covers the period of cell preparation for the replication process
and the process itself which usually happens 2–6 times. At this stage, growth
intensity is not too high, because it is based on the increase in the number of cells
and less on their volume increase. During this phase accumulation of organic matter
The biology of plant growth and development includes the fundamental analysis of
the mechanisms and phenomena that underlie cellular differentiation, intra- and
intercellular interaction within the entire organism to form cells, tissues and spe-
cialized bodies. Plants, unlike animals, have a reversible plastic morphogenetic
process. Although the number of structural genes in plants and animals is almost the
same, these kingdoms differ substantially by the number of specialized cell types
and tissues. Thus, in plants were identified only a few tens of types of tissues and 60
different types of specialized cells, while in vertebrates the number of the latter is
several hundreds.
Structural and morphological differences between plants and animals are
determined by morphogenesis and development mechanisms, which were formed
during the evolution and ensured the phylogenetic separation of these kingdoms.
The primary role in the occurrence of certain features of morphogenesis is owed to
the fact that plants are fixed to the substrate, so that growth and development is
dependent on and is in direct connection with varying environmental factors.
Development in plants and animals has a cyclic character. In plant organisms the
succession of generations occurs—sporophyte–gametophyte (haploid-diploid or
sexual-sexual). During individual development highly specialized complex struc-
tures like the flowers and the fruits form from a practically undifferentiated embryo.
At the cellular level, the formation of different morphological structures occurs
through continuous functional activity of undifferentiated meristems. The rigid
cellulose-pectin cell walls exclude cell migration during morphogenesis. Plant cells
are omnipotent—a unique feature, determined by the plasticity of the plant genome,
which allows in certain circumstances, even partially differentiated cells to switch to
another program of morphogenetic development and to even ensure regeneration of
the entire organism. In animals stem cells etc. are similar to meristematic tissues,
but only embryonic stem cells have a comparable plasticity.
In terms of molecular biology, the development process represents differential
gene activation and suppression. Inclusion of each new developmental program is
reflected in the modified spectrum of gene expression products—mRNAs and
proteins. Development of modern research methods of nucleic acids and proteins
198 7 Plant Growth and Development
allowed to determine the mechanisms of temporal and spatial gene regulation which
contributes to the elucidation of plant growth and development. It was found that
only about 4 % of the polypeptides are organ-specific, while the mRNA level of
specificity is much higher—25 %. An important issue is finding the nature of these
differences and the role of specific proteins in inducing morphogenetic programs
and organ differentiation. It’s possible that these key proteins are synthesized in
small quantities during short time periods.
A key direction in the study of plant growth and development is the isolation and
functional analysis of specific genes with an important role in morphogenesis. In
this regard the creation of cDNA libraries (Fig. 7.10) and their subsequent
hybridization with DNA or RNA enables the identification of specific active genes
for different tissues or organs at different stages of vegetation.
Plant growth and development are complex processes conditioned by three key
types of factors—nutritional, genetic and hormonal.
In 1675 the Italian scientist Marcello Malpighi predicted that plants contain
substances with regulatory effect. First experiences that have shown the presence of
stimuli acting on the movement of plants were exposed by Francis Darwin and his
200 7 Plant Growth and Development
father, Charles Darwin (1880) in his famous On the Movements and Habits of
Climbing Plants.
The first scientist who worked with hormones and active extracts from plants
and introduced the concept of “hormone” in plant physiology was Fitting (1909–
1910). Phytohormones (from gr. phyton “plant” and hormaein “to stimulate, to
excite”) are natural organic compounds with relatively low molarity and varied
chemical structure, participating in the interdependent activation of cells, tissues
and organs and required in small quantities (10−6–10−11) for activation and
implementation of physiological programs. They are synthesized in specialized
tissues of higher plants and transported throughout the plant body coordinating
ontogenesis, stimulating or inhibiting the morphogenetic pathways or altering the
quantity and quality of the essential processes in the organism.
In contrast with animals that have special glands that synthesize hormones, their
biosynthesis in plants takes place mainly in the meristematic tissues of growth
centers, phytohormones are multivalent and polyfunctional (Fig. 7.11).
Traditionally phytohormones are divided into 5 groups (Table 7.1):
• auxins (derivatives of the indole-3-acetic acid);
• cytokinins (derivatives of 6-aminopurine-zeatin);
• gibberellins (tetracyclic carbonic acids of the diterpenoid class-GA3);
• abscisins (abscisic acid ABA—a sesquiterpenoid with optical activity);
• ethylene (colorless gas, unsaturated hydrocarbon with a double bond).
Similar actions have some oligosaccharides—salicylic acid, polyamines.
Brassinosteroids are also considered hormones (with stimulatory action), as well
as fusicoccins and anthesins. Novel compounds similar to cytokinins—4-phenilthio
redo salicylic acid and 1-(3-chlorophenyl)-3-(2-pyridyl) urea are also being tested.
Between different groups of hormones there are synergistic relationships (auxins
stimulate gibberellins) and antagonistic ones (between abscisic acid and auxins).
Fig. 7.11 The diagram shows gradual changes in the location of certain regions of the leaf (blue
dots) and the concentration (the size of blue dots) of free IAA production during the development
of the leaf primordium in Arabidopsis. Arrows indicate locations with the highest level of primary
production of free auxin located on the margins of the leaf blade at each stage of development
(a–d) while short arrows indicate the location of reduced auxin synthesis rates (d, e)
7.6 Endogenous Factors of Plant Growth and Development 201
Natural and synthetic substances that act as growth regulators, according to the
mode of action, are divided into (Fig. 7.12):
• growth promoters;
• growth inhibitors;
• retardants (only synthetic).
Auxins, gibberellins and cytokinins are considered stimulants and abscisins and
ethylene—inhibitory hormones. There are multiple links between them, they have a
versatile action, which depends, on one hand, on the concentration of the hormone
that has reached the target cells, on the other hand, on the tissue competence (ability
to respond, type and intensity of the response) (Fig. 7.13). These three groups of
202 7 Plant Growth and Development
substances are not acting separately. They interact in such a manner that all stages
of growth and development are the result of an equilibrium between stimulators and
inhibitors that manifest mainly in seasonal processes.
Growth stimulants (auxins, gibberellins, cytokinins, substances of the vitamin B
group, ethylene chlorohydrin, thiourea, etc.) are organic substances that stimulate
endogenous plant morphogenesis and regulate physiological correlations between
different organs of the plant. If a paste containing vitamins is applied on the growth
cone or at the basis of a leaf, we would see increased growth, the formation of
shoots and fruit components. Stimulants are used in agriculture to facilitate the
rooting of certain cuttings, in order to increase plant productivity.
Growth inhibitors (unsaturated lactones, phenolic compounds, organic acids and
flavonoids) are endogenous organic substances, which inhibit the plant physio-
logical activity and the development of organs. Growth inhibitors are present in
different organs of the plant—seeds, bulbs, tubers, shoots and reduce or cancel the
activity of stimulants and inhibit plant growth, seed germination, alter the activity of
enzymes, inducing deep dormancy. Their content increases maximally in autumn
during the transition to the dormant state, which is related to interruption of mer-
istematic tissue growth. Among natural inhibitors are: the β inhibitor, derived from
stems and roots, which inhibits seed germination inside the fruit, the abscisic acid,
derived from dormant buds that has multiple properties (anti-stimulating action,
maintenance of the repose state, young fruit detachment etc.); phlorizin is synthe-
sized only in leaves only during short days and inhibits the respiration process,
synthesis of nucleic acids and proteins; coumarin inhibits seed germination etc.
Fungicides, nematicides, insecticides and other groups of toxic substances used
in agriculture, also act by inhibiting important vital processes.
7.6 Endogenous Factors of Plant Growth and Development 203
The inhibitory effect can be noticed also during the action of physical factors
(cold or extreme weather, light excess or shortage, deficit or excess of moisture) and
chemical factors (chloropropane, butylate and alachlor impede germination, atra-
zine, simazine, propazine block photosynthesis).
Retardants are artificial substances retaining plant growth and development.
7.6.1 Auxins
ARF is a transcription factor that can enter the nucleus, where it binds the
promoter sequences of various genes and alters gene expression levels. IAA
influences polyribosomes and the activity of the nuclear apparatus including:
• RNA polymerase (RNA polymerase-1) due to the increasing content of the
transcription initiation factor ϒ;
206 7 Plant Growth and Development
Fig. 7.16 Auxin location revealed by immunostaining and viewed with a confocal microscope
(a) and expression of the genes DR5:GUS in transgenic Arabidopsis thaliana, demonstrating
histochemical localization of GUS activity during morphogenesis of leaf primordia (b–f).
a Lamina paradermal section with IAA strong marking (characteristic) (green-fluorescent staining
by secondary antibody conjugation) in chloroplasts (indicated by arrows) and a lower
concentration inside the cytoplasm of elongated cells near the fibro-vascular bundle. b Strong
expression of the GUS gene in stipules (marked with arrows), the promeristem without the GUS
gene (marked with a large arrow), foliar primordium without the GUS gene (marked with a short
arrow) and early production (low expression of the GUS gene) without IAA production (marked
with a large short arrow) in the tip of a leaf primordium. c Gene expression in all active
hydathodes (two are marked by arrows) of the leaf primordia. d Strong expression of the GUS
gene in a marginal hydathode during development (marked with arrows) and a fibro-vascular
bundle in differentiation (indicated by short arrows) with low activity of the GUS gene. e GUS
gene expression at the base of trichomes (arrows) and two venules ending free (short arrows)
associated with trichomes. f Reporter gene expression (marked with a short arrow) at the top of the
venule that ends in the primordium of the developing lamina
7.6.2 Gibberellins
Gibberellins are named after the fungus Gibberella fujikuroi, in which they have
been identified for the first time (1926). From the chemical point of view, they are
tetracycle diterpenoids. The symbol used for gibberellin notation is GA, equipped
with a numeric index starting with 1 (GA1, GA2, GA3, …). There have been
identified over 70 types of gibberellins, gibberellic acid 3 (GA3) is considered more
active. Gibberellins are found in the free state and bound with glycosides.
fertile line and its appearance after application of gibberellins GA3 tells about the
induced expression in the fertile line of a gene similar to orfH522 associated with
cytoplasmic male sterility (CMS) in sunflower.
Sequencing and comparison with the nucleotide sequence of orfH522 present in the
EMBL database (EMBL/GenBank Accession X55963) showed 99–100 % homology
with the mitochondrial gene sequence orfH522 selected for amplification (Fig. 7.19).
Consequently, the exogenous application of GA3 during budding induces the de
novo synthesis of a protein compound with the relative molecular weight of 16 kDa,
7.6 Endogenous Factors of Plant Growth and Development 209
Fig. 7.18 RT-PCR products obtained with primers specific for orfH522 (a) and actin (b). Total
RNA was extracted from calathidium of fertile SW501 (F) and sterile SW501CMS (S) plants
treated and untreated with GA3 (“+” “–”), M—marker 100 bp and 1 kb. (Duca et al. 2006)
Fig. 7.19 The nucleotide sequence of the sunflower mitochondrial orfH522 and the amplified
sequence (I)
similar to that identified in CSM lines (Fig. 7.20). These results support the idea that
GA3, causes male sterility by inducing the expression of a similar open reading
frame, because the translation products are also identical.
Biological significance. Physiological effects of gibberellins on plants are mul-
tilateral. Among the most important functions can be mentioned:
• stimulates stem elongation in dwarf plants, so many dwarfism genes are gib-
berellin deficiency genes;
• accelerates flowering in long day plants;
• stimulates caryopsis germination in cereals, stimulates fruit growth;
• determines changes in the photoperiod;
• intervenes in ceasing bud dormancy. Brings seeds out of the dormant state and
influences their germination by intensifying the formation of ribosomes and
nucleic acids, but also by permeating membranes;
• in the endosperm gibberellins are participating in endoplasmic reticulum
development, cell wall degradation and synthesis of a large number of hydro-
lytic enzymes that catabolize seed reserves and the formed metabolites ensure
embryo and seedling development (Fig. 7.21);
210 7 Plant Growth and Development
Fig. 7.20 Electrophoresis of total protein pools from the leaves (a) and inflorescences (b) of
fertile sunflower plants SW501 (F) and sterile SWS01ASC (S) treated and untreated with GA3;
M—marker (10–100 kDa); “+”—variants treated with GA3. (Duca et al. 2006)
• among the enzymes induced by gibberellins are α-amylase, some proteases, acid
phosphatase, β-gluconase, α-glucosidase and ribonuclease;
• determines the sex switch in plants, causes parthenocarpy;
• intensifies transpiration, photosynthesis and respiration;
• shows synergism with auxins, due to its action on auxinoxidases;
• by stimulating cell division, gibberellins control mitotic activity, activate
enzymes responsible for phospholipid biosynthesis.
Practical applications. Based on these properties, gibberellins have wide
application in practice—they are used to stimulate tomato fruit formation, to stop
dormancy in tubers, buds, seeds, etc.
7.6.3 Cytokinins
Fig. 7.21 The role of gibberellins in seed germination. 1 GA induces water imbibition of the
embryo; 2 GA stimulates the production of α-amylase by aleurone cells; 3 Amylase cleaves starch
reserves from the endosperm; 4 Carbohydrates fuel the growth processes of the embryo
Fig. 7.22 The gene for cytokinin biosynthesis. a Chloroplasts from seeds grown in the dark
without cytokinins; b Chloroplasts from seeds grown in the dark with cytokinins, notice the
formation of thylakoids
• mobilize and attract metabolic substances and minerals to the dominant centers;
• stimulate the exit from dormancy and neutralize apical dominance;
• activate callogenesis as a result of active cell division;
• activate differentiation of adventitious buds on stems and roots;
• involved in apical dominance, stimulate seed germination and induce anthesis;
• increase resistance to cold and toxic substances;
7.6 Endogenous Factors of Plant Growth and Development 213
Has the ability to form glycoside by interacting with glucose, thus representing
an inactive non-toxic form. ABA is also inactivated by hydroxylation in the
endoplasmic reticulum.
Biosynthesis. Is synthesized in mature leaves and fruits. ABA biosynthesis
occurs via two pathways (Fig. 7.24):
• from mevalonic acid → isopentenyl pyrophosphate → heranylpirophosphate;
• by carotenoid and violaxanthine decomposition → xanthoxin → abscisic acid.
It was found that induction of ABA synthesis occurs during genome repro-
gramming and synthesis of increased amounts of ABA-inducing polypeptides, of
which lectins are more significant (especially agglutinins in wheat).
Transport. ABA is transported from its synthesis location through the phloem
and xylem to the parenchyma, the descendent flow being three times greater than
the ascendant.
Mechanism of action. The primary action is carried in the membranes of target
cells which contain protein receptors specific for ABA, influencing the lipid phase
214 7 Plant Growth and Development
7.6.5 Ethylene
Ethylene (H2C = CH2)—a gas that is formed in plant organs, being an inhibiting
hormone and an auxin antagonist. It was found in plants and fungi and does not
216 7 Plant Growth and Development
Fig. 7.26 The axes of incubation depending on ABA concentration (Ried et al. 1990). The axes of
hibernating and non-hibernating wheat seeds incubated for 28 h at 20 °C on agar-agar containing
varying concentrations of ABA
occur in bacteria, algae, and animals. It is considered as the maturation and aging
hormone, since it stimulates apoptosis. At low concentrations (0.04–1.0 μl) it shows
strong morphogenetic effects. It was described in 1901 by Neliubov, which dem-
onstrated the role of ethylene in inhibiting stem elongation, thickening and hori-
zontal orientation. In the 20s of XXth century it was found that ethylene accelerates
fruit ripening.
Biosynthesis. Ethylene in higher plants is synthesized in the presence of light,
from methionine, with 1 amino cyclopropan-1-carbonate (ACC) as precursor,
which can serve as a transportation form.
Transport. Ethylene diffuses freely through the intercellular spaces.
Mechanism of action. Ethylene action on biological processes is very fast, which
leads to the idea that like other phytohormones, ethylene has a signaling role.
Biological significance (Fig. 7.27).
• causes epinasties;
• delays growth and development;
• stimulates organ aging;
• accelerates fruit ripening and their detachment (Fig. 7.28);
• contributes to flower, bud and leaf detachment;
• blocks the growth of leaves, plant elongation, mitosis;
• slows down polar auxin transport and removes apical dominance;
• switches the direction of growth form the longitudinal one to the transversal one,
contributing to thickening of the stems;
7.6 Endogenous Factors of Plant Growth and Development 217
Fig. 7.27 The role of ethylene in various physiological processes. (I) Germinating seed of
Arobidopsis at 4 °C for 4 days and then transferred to dark for 72 h; a Wild-type genotype grown
in the presence of ethylene at 10 μl/l; b Wild-type genotype grown without ethylene (Guzman
et al. 1990); (II) Leaf detachment
Fig. 7.28 Involvement of ethylene in fruit ripening. (I) Model of ethylene receptor action. In the
absence of ethylene, receptors (AR) actively suppress ethylene responses and fruit ripening. Before
binding ethylene receptors become inactive (IR) and ethylene responses can be initiated. Mutated
receptors (M) cannot bind ethylene and continue to actively suppress ethylene responses (Klee
2004). (II) Induction of fruit ripening in the presence of ethylene. (III) Lack of the ethylene
receptor gene in tomatoes on the left
218 7 Plant Growth and Development
Fig. 7.30 Examples of plants that require vernalization (Amasmo 2004). a Brassica oleracea,
b Hyoscyamus niger, c Arabidopsis
Other external factors affect plant growth and development and the speed of growth
depends on the intensity of all physiological processes—nutrition (carbon dioxide
and nutrients), water supply, energy and substance exchange.
Temperature. Plant growth is possible in different temperature amplitudes. Early
spring plants grow even at temperatures below 0 °C (wheat, peas at −20 °C). There
are plants which start their growth at the limit of +50 °C. For each species,
according to its characteristics and in particular, the geographical origin there are
specific temperature limits within which they can grow and develop. For each
species there are three important temperatures:
(1) The minimum—which starts growth;
(2) The optimal—providing the most favorable conditions;
(3) The maximal—which stops growth.
It was established that plants grow very intense during night. For many plants,
alternating temperatures are favorable: very high during the day and low at night.
This process was named by F. Vent, thermoperiodism.
Mineral nutrition. The increase is significantly influenced by the amount of
nutrients in the soil and, in particular, by the concentration of nitrogen. A high
concentration of nitrogen, however, stimulates plant rapid growth, inhibits differ-
entiation of organs, such as flower formation and possibly, blooming. A rich
background of minerals contributes to the formation of abundant green mass, which
is favorable for forage cultivation (so, fertilizers lead to a decrease in the abundance
of fruit and seeds).
The amount of water. During the process of growth an invaluable importance has
the water supply. Lowering the quantity of ground water leads to lower concen-
tration of water in plants, and this in turn leads to slow growth—reduced rate of cell
division and, in particular, elongation.
Light influences intensity and character of growth. Photosynthesis and organic
mass accumulation occurs more intense in daylight and cell elongation is more
intense in the dark. Light has a great influence on the process of organ formation. In
plants grown in the dark chlorophyll “b” is not synthesized, so they have a yellowish
tint and are called etiolated. Etiolated plantlets are characterized by a number of
anatomical and morphological features—a simplification of the anatomy of the stem
—there is a weak development of central cylinder tissues and mechanical tissues,
therefore long thin stems form; leaves are reduced in size. Stem and root elongation
in etiolated seedlings appeared during the evolution process because, in most cases,
seed growth occurs in the soil, in absence of light and the lack of leaves and other
mentioned peculiarities facilitate seedling passage through the soil.
It is possible that stem elongation in the absence of light is due to the lack of
growth inhibitors. During the dark phase many hormones, auxins. Disturbance of
the auxins/growth inhibitors ratio causes irregular growth.
222 7 Plant Growth and Development
Once the seedlings grow through the soil surface, internal and external changes
occur. In the dark, in dicotyledonous plants, hypocotyl is upside down and protects
the growing point from damage. Under the action of light it is straightening. In light
stem growth slows down, leaf growth accelerates and they acquire their regular
shape. Under the action of light the stem epidermis forms.
These changes are caused by red light with a wavelength of 660 nm absorbed by
the phytochrome pigment, which is a chromoprotein with molecular weight of
120,000 kDa. Phytochromes which absorb red rays are called red phytochromes
(Pr), and those that absorb red rays with the wavelength of 730 nm, are called long-
wave red phytochroms (Prf).
Phytocromes can be found in different plant organs, being involved in photo-
morphogenetic programs, including photoperiodic response in plants.
All the processes controlled by the phytochrome system can be classified into
two groups:
• processes that under the influence of red light intensify—differentiation of the
epidermis, anthocyan biosynthesis, seed germination;
• processes that are inhibited—stem growth, hypocotyl elongation.
The phytochrome system is evolutionary very old, and is also encountered in
blue-green algae and heterotrophic organisms.
Oxygen nutrition. Growing process require a lot of energy resulting from the
process of respiration. Therefore the influence of O2 reflects on growth. Reducing
the amount of O2 under 5 % retains growth. This is the result not only of disturbed
energy balance, but also of the accumulation of products of anaerobic exchange
(alcohol, lactic acid).
Excitability and movement, which are essential qualities of living matter, are also
present in plants.
Various movements can be found in plants and they are based on the changes in
the turgidity of different groups of cells and tissues.
Plant growth movements are known from ancient times, being described by
Teofrast that detected movements in Mimosa pudica and clover plants.
Physiological mechanisms of growth movements have been explained by the
1960–1970 years, when Holodnâi and Vent developed the hormone theory of
growth movements. At the basis of these mechanisms stay changes in phytohor-
mone content.
Plant movements are divided into:
• passive;
• active.
7.9 Plant Growth Movements—Tropism and Nasties 223
Passive movements are favored by the presence of adaptations that allow entire
plants or parts of their bodies to be moved by physical or biological agents from one
place to another. They are produced by metabolic energy expenditure, as deter-
mined by physical and chemical factors (movement of pollen, seeds, etc.).
Active movements are made by plants with the use of their own energy.
Regardless of the structure in which they occur (inside plant cells or organs), active
movements can be autonomous or induced.
Autonomous movements are executed under the influence of internal factors,
specific to the organism. These movements in higher plants are known as nutations.
Induced movements are determined by the direction of action and intensity
variations of some external environmental factors.
Induced movements performed by the organs of fixed plants are called tropisms
while the ones executed by plant organs under the influence of intensity variation of
environmental factors are called nastic movements.
At the cellular level both autonomous and induced movements are present.
Among these movements ciclose movements, cytoplasmic currents and chloroplast
movements can be highlighted.
Active movements are accompanied by ATP and metabolic energy expenditure.
Based on the above mentioned, we can deduce:
• intracellular movements—movements of fixed plants (tropisms, nastic
movements);
• intercellular movements—movements of cytoplasm, chloroplasts, the nucleus.
Tropisms are movements related to the unilateral direct action of a factor (light,
gravitation). These are directed growth movements towards the stimulant.
Movements oriented to excitation source are called positive movements, while
those away from this source—negative movements.
Depending on the stimulating factors that influence the physiological and bio-
chemical processes from the protoplasm, tropisms are divided into:
• geotropisms—caused by unilateral and directed action of the earth gravitational
force. Stem growth via its apex is negative geotropism, root growth—positive
geotropism;
• phototropisms—caused by the unilateral action of light. Stem growth is positive
phototropism and root growth—negative phototropism;
• seismotropisms—the movement of plants towards an object (vines that cling to
other plants, for example, beans) (Fig. 7.31);
• chemotropisms—root growth towards the source of granular fertilizers, pollen
tube growth through the stigma to the ovary (Fig. 7.32);
• hydrotropisms—growth towards the source of water in the soil;
• electrotropisms—passage of electric current through the organism of a plant
induces bending towards the positive pole.
224 7 Plant Growth and Development
Fig. 7.32 Pollen movement. Cytoarhitecture is maintained after treatment with incompatible
protein-S. Images were recorded in the apical pollen tube after 1 min. (a) after 4 min. (b) after
6 min. (c) after 8 min. (d) of -S protein induction
Nastic movements represent growth movements that are determined by the dif-
fuse action of excitatory factors. They are observed in plants with dorsoventral
symmetry and cause two types of movements: epinasties—bending downwards and
hyponasties—bending upwards (Fig. 7.33).
7.9 Plant Growth Movements—Tropism and Nasties 225
If growth is faster at the top of the organs, then movements are called epinasties.
Such kind of movements cause floral buds to open. If the increase is more intense in
the organs, the movements are called hyponastic movements.
Depending on the factor causing nastic movements, we distinguish:
• nyctinastic movements;
• seismonastic movements;
• termonastic movements.
Nyctinastic movements are determined by the alternation of day and night.
These are movements of flowers and leaves caused by changes in temperature and
light. Seismonastic movement are the phenomenon of closure and opening of the
petals in Mimosa pudica induced by touch. Termonastic movements represent
growth movement caused by the diffuse action of temperature, opening and closure
of tulip flowers depending on temperature.
Plants are integral organisms and consist of 50–60 cell types, 12–15 types of tissues
and 5–6 organs and specialized organ systems. Although they differ by their
structure and functions during growth and development, they interact through
conducting vessels forming a whole. In the growth and development processes
regulation and self-regulation of all physiological processes occurs which ensures
homeostasis (morphological, biochemical, genetic).
Self-regulation represents all the systems and mechanisms that help maintain
homeostasis in the fluctuating environmental conditions at the cellular level, based
on the interaction of the:
• genetic system;
• enzymatic system;
• membrane system.
226 7 Plant Growth and Development
After further division of the egg (the division plane is perpendicular to the
polarization axis), daughter nuclei have different conditions which create the
polarized cytoplasm. As a consequence, different genetic programs and differenti-
ation paths may be established.
The direction of cell polarization changes continuously along embryogenesis,
during the formation of primordia in the apex, during the process of hystogenesis in
the leaves, root formation in the pericycle.
Each meristematic polarized cell can divide into two different or identical cells,
in the first case there is division of differentiation, and in the second—one of
multiplication.
An important role in creating polarity in higher plants have phytohormones
which create concentration gradients in various plant organs. Heteroauxin and its
physiological analogues β-indolyl acetic acid and α-naphthylacetic acid, substances
which actively influence growth and development of the root. Acropetal gibberellic
acid acts on the acropetal current leading to the stimulation of sprouting and
development of shoots. Auxins stimulate cell growth in areas of the basal shoots
and gibberellins stimulate apical bud areas.
IAA, by circulating in a polar manner, accumulates at the bottom end which
induces morphological and genetic programs of root formation. Thus, in plants,
opposite ends of the axis differ essentially, for instance in grapevine cuttings by the
formation of roots on the morphological bottom and of shoots—from buds exposed
on the morphological top.
Phytohormones do not act directly on basipetal and acropetal currents of sub-
stances. They lead to the intensification of growth processes in the locations where
they are applied and in this manner the ability forms in these centers to attract
substances that contribute to the polarization of the plant body.
Biopotentials in higher plants are polar and are the expression of metabolic
gradients between dominant centers—apical and root portions corresponding to the
transport of water and mineral salts. In a leaf, the apical electrode is usually positive
compared to the basal electrode (≈100 mV), and the difference can also show
variations between day and night. The inner side of the leaf is negative compared to
the top surface; between the illuminated zone of a leaf and a zone which stays in
shadow, a potential difference of 50–100 mV appears in a few minutes (the shaded
portion is negative). Periods of light and dark can be alternated several times with
the same result.
Along onion root cells there are dipoles placed in series. The root tip is positive
compared to the middle part and they are negative in comparison to the core. The
magnitude is of the order of tens of millivolts. Probably in the middle zone of the
root cell dipoles change orientation (along the roots of beans spontaneous potential
oscillations were revealed with small amplitude (<2.5 mV) and with a period of a
few minutes). They are due to fluctuations in time of the currents of Na+ and K+, as
a result of self-regulation of the cellular osmolarity.
228 7 Plant Growth and Development
Glossary
References
Amasmo R (2004) Vernalisation, competence, and the epigenetic memory of winter. Plant Cell
16:2553–2559
Chaylahyan MX et al (1962) Terminologiya rosta i razvitiya vysshih rasteniy, 96 pp
Duca M, Port A, Orozco-Cardenas ML, Lovatt C (2006) Mecanisme moleculare ale
androsterilității ereditare şi induse la floarea-soarelui. Buletinul AŞM 298(1):86–93
Gilbert SF (2000) Developmental Biology, 6th edn. 709 pp
Guzman P, Ecker JR (1990) Exploiting the triple response of Arabidopsis to identify ethylene-
related mutants. Plant Cell 2:513–523
Ivanov BB (1974) Kletochnye osnovy rosta rasteniy, 223 pp
References 229
Historical Background
Brief Updates
Biorhythms are one of the ways in which organism integrity is manifested, rep-
resenting a periodic fluctuation within a range of certain biological processes or
phenomena, a controlled chronobiological variation. The notion of rhythm includes
the entire set of processes and transformations based on self-regulating mechanisms
and systems that contribute to a harmony, a strict organization of events that occur
in nature.
Biological rhythms are characteristic of all levels of organization of the living
matter—from the molecular and submolecular to the biospheric, which shows that
biorhythmicity is a universal property of living systems.
Biorhythms are recognized as adjustment mechanisms that maintain body
homeostasis, dynamic equilibrium and adaptive processes in living systems.
Biological rhythms are endogenous, being determined and regulated by genes, but
are also closely related to external environmental factors.
The temporal organization of living systems happened during the evolution of
the organic world in a certain manner. Biological rhythms are found in all
homeostatic control systems, allowing organisms to behave as complex entities,
integrated in the multitude of influences exerted by the external environment:
diurnal, monthly, seasonal. Particular attention should be paid to the fact that the
very existence of rhythmic changes in nature contributed to the evolution of
organisms, being reflected in certain aspects of phylogenesis.
One of the most important factors that dictates periodicity phenomena is the
rotation of the Earth around its axis and around the sun which reflects on the
succession of light/dark cycles, weather, droughts, floods etc. Besides this con-
tributing to the rhythmicity of life in ecosystems can be the seismic and volcanic
activity, or other local phenomena. Altogether, these factors create a rhythm which
is strongly intertwined with the life cycle of an organism or species.
Over the years scientists have been concerned with the idea of classifying bio-
logical rhythms.
234 8 Plant Biorhythms
Fig. 8.1 The circadian system in Arabidopsis and its relationship with the CO flowering gene
(Hayama and Coupland 2004). The model of the circadian clock is as follows. Phytochromes and
cryptochromes receive light and are involved in resetting the circadian horologe. ELF3 and ZTL
are intermediaries between the photoreceptors and the circadian clock. TOC1/ELF4 and LHY/
CCA1 form a negative inverse relationship with circadian oscillators. LHY/CCA1 put in action
negative regulators for TOC1 and ELF4 which positively regulate LHY/CCA1 transcription.
Oscillating functions determine the transcription stage of CO, which is a key intermediary gene
between the circadian clock and flowering. CO transcription is regulated by FKF1 and GI, whose
transcription is controlled by the circadian clock. The FKF1 protein is controlled directly by light,
and this fact allows to increase CO transcription during long day conditions. The CO protein is
also directly activated by light, and this allows CO to generate a long day signal and to activate the
flowering time gene—FT to promote flowering especially in conditions of long days. Circadian
Clock Associated 1 (CCA1), Late elongated hypocotyl (LHY), Timing of Cab Expression 1
(TOC1), Early Flowering 4 (ELF4), Early Flowering 3 (ELF3), Zeitlupe (ZTL), Flowering-time
genes Constants (CO), Flowering Locus T (FT), Flavin binding, Kelch Repeat, F-Box (FKF1),
Gigantea gene (Gl)
236 8 Plant Biorhythms
A characteristic property for plants besides the bipolar axes system is the typical
alternation of nodes with internodes, of leafs with lateral buds etc. These metamers
develop consecutively and determine a structure that optimizes the photosynthetic
activity of leaves. It should be mentioned that plants are characterized by a certain
rate by which they generate new leaves, nodes and internodes and which has been
called the plastochron, so that the physiological rhythm in the apical bud of the
plant (where the meristematic tissue is located) turns into a spatial pattern.
For instance, the presence of oscillatory processes can be observed in the
structure of the plant corm. Presence of regularity can be noticed in the layered
arrangement of the starch grains, cell walls and annual rings, in the very compli-
cated configuration of leafs, flowers, in the complex structure of the inflorescences
etc. (Fig. 8.2). It is assumed that, in sunflower, oscillations with a certain periodicity
are the phenomenon responsible for the helical arrangement of leaves on the stem
and the helical arrangement of flowers on the calathidium.
After the periodical activity of the cambium, which in temperate zones is a
seasonal phenomenon, concentric layers are forming in the secondary phloem and
xylem, which correspond to a year and are called annual rings. The width of these
layers varies depending on the climatic conditions for that particular year, namely,
the amount of light, temperature, rainfall, soil moisture, the duration of the growth
season etc.
8.2 Biological Rhythms in Plants 237
Fig. 8.4 The photoperiodic characteristic of flowering (Eriksson and Millar 2003)
• short-day plants, which flower also in the presence of a dark period longer than
the critical period of day/night alternation (Fig. 8.4);
• plants, which, in order to reach the reproduction stage, need a long day duration,
then a short one;
• plants that are flowering faster if they first pass through a period of short days,
followed by long days.
There is also the qualitative photoperiodic response of plants with photoperiodic
control, which determines the flowering of plants (short-day or long-day) in
unfavorable lighting conditions.
The photoperiodic response can be suddenly altered under the influence of other
environmental factors. It should be noted that photoperiodism which influences
plant growth, ultimately regulates also the intensity of photosynthesis.
Thus, the role of the circadian system in the signaling network is that of an
interface between the environmental factors and the internal programs. The circa-
dian clock evolved as an adaptation to the 24h rotation of the Earth around its axis
and the fluctuations of light intensity and temperature that accompany it.
Photoautotrophic organisms must be exposed to sunlight in order to carry photo-
synthesis. Thus, all plants are subjected to the day/night cycle, except for the
seedlings that germinate in the soil or the seedlings that grow and develop in the
subpolar regions of the globe. The circadian system allows organisms to anticipate
these cycles by detecting light or heat levels. The 24h biological rhythm charac-
terizing the circadian clock is not a direct response to external factors and persists
even in constant environmental conditions, with a periodicity that often differs from
24 h. The circadian system retains information from previous days. Rhythmic
processes are similar for all organisms. Molecular research on circadian rhythms in
plants have advanced mostly in Arabidopsis thaliana, although this physiological
phenomenon was also investigated in other species, among which could be men-
tioned Ipomoea nil(Pharbitis nil), Kolanchoe and Phaseolus.
The circadian rhythm is characterized by the following parameters: an endog-
enous duration of approximately 24 h (the fluctuation period), a quantitative extent
in the fluctuation of the parameters (amplitude) which is also dependent on external
stimuli, the dependence of the period on temperature, the presence of rhythmicity in
the absence of the input signal (Fig. 8.5).
Microarray analyzes showed that at least 6 % of Arabidopsis genes are expressed
rhythmically, with expression peaks at all stages throughout the day and night.
Gene expression patterns produce an impact on the physiological processes in
plants, some of which are obvious (like “sleep movements” of leaves in legumes),
while others less so. In many cases, the genes that affect a particular pathway or
process are expressed during the same phase. Multiple genes encoding the bio-
synthesis of phenylpropanoid enzymes have maximum RNA levels before dawn,
Fig. 8.6 The scheme of the circadian rhythm in Arabidopsis (Onai and Ishiura 2005)
of LHY, CCA1 and TOC1 and profoundly affect the circadian system, but they
have not yet been cloned and sequenced.
Nature provides a complex set of signals during the day/night cycle, including
changes in temperature, light quantity and quality. Thus, the combination of signals
during the day/night cycle reinforces the precision of biological clock functioning
and maintain its correct period. If the circadian clock was lagging behind, the
biological processes would occur later than during certain optimal environmental
conditions. By contrast, a faster circadian clock would be initiating the rhythmic
processes too early.
A constant signal, for instance a short light pulse transiently affects oscillatory
components and, subsequently, the length of the period resulting in either clock
acceleration or delay.
8.4.2 Temperature
Circadian rhythms are also reflected in varying temperatures during the day/night
cycles. The role of temperature in manifestation of the biological rhythms is elo-
quently represented in Arabidopsis, where, depending on the day or night, tem-
perature values have a difference of only 4 °C or even less.
8.4 The Molecular Mechanism of the Circadian Clock 243
8.4.3 Light
Exposure of the plants to light affects one of the oscillatory components, which is to
reactivate the clock. Several target-mechanisms involved in the initiation of bio-
logical rhythms have been described in plants.
The first mechanism can be determined by the involvement of PIF3 (phyto-
chrome-interacting factor 3). PIF3 is a protein that binds to DNA, including to the
G-box sequence of the CCA1 and LHY promoters, which is present in several
genes activated by light.
In enacting biological rhythms in plants proteins from the ZEITLUPE (ZTL),
Flavin-binding-Kelch-F-box (FKF) and LOV-Kelch 2 (LKP2) families might also
be involved. They contain a domain related to Period-ARNT-Sim (PAS), which
binds with the flavin chromophore in phototropin photoreceptors. ZTL mutant
phenotypes are light dependent, suggesting a possible photoreceptor role. The ZTL
protein has the potential to interact both with phyB, and cry1, which may cause
indirect dependence on light.
244 8 Plant Biorhythms
Fig. 8.7 Induction of flowering in Arabidopsis depending on day length (long day and short day)
(Hayama and Coupland 2004)
Glossary
References
Hayama R, Coupland G (2004) The molecular basis of diversity in the photoperiodic flowering
responses of arabidopsis and rice. Plant Physiol 135:677–684
Makarov V, Poznyakova V (1989) Nauka o biologicheskih ritmah, sostoyanie, problemy,
perspektivy. Biologiya v shkole, nr. 6, pp 5–10
Melnic (1985) Metronomul biologic. Chişinău
Moore-Ede MC, Sulzman PM, Fuller CA (1982) The clocks that time us. Harvard University
Press, Cambridge
Naumov SP (1989) Zoologia vertebratelor. Chişinău, Lumina
Onai К, Ishiura M (2005) PHYTOCLOCK 1 encoding a novel GARP protein essential for the
Arabidopsis circadian clock. Genes Cells 10:963–972
Polevoy VV (1982) Fitogormony. Leningrad, pp 69–74
Polevoy VV, Salamatova TS (1991) Fiziologiya rosta i razvitiya rasteniy. Leningrad, pp 158–161
Reyvn P, Jevert R, Aykhorn S (1990) Sovremennaya botanika, vol 2. M. Mir, pp 119–123
Somers DE (1999) The physiology and molecular bases of the plant circadian clock. Plant Physiol
121:9–19
Chapter 9
Elimination of Substances in Plants
Historical Background
Brief Updates
Today several hundred species of insectivorous plants (carnivorous) are known.
Similar to other typical green plants, carnivorous plants have organs required for
normal nutrition. Thus, they possess green leaves that can absorb CO2 from the air
and the process of photosynthesis takes place mostly according to the C3 type. This
type of nutrition in carnivorous plants was caused by low nitrogen content in the
soil. It is considered that nitrogen compounds extracted by insectivorous plants
from captured prey significantly enhance photosynthesis. In some representatives,
the leaves have a completely altered shape while in others they have only partially
transformed in specialized organs designed to capture insects and possess secreting
glands or other aromatic substances. Carnivorous plants are divided into three
9 Elimination of Substances in Plants 249
groups, based on the adaptations they have developed during evolution that allow
capturing the prey. The first group includes plants that capture prey with sticky
leaves. The second group is comprised of plants that have traps in the form of
vessels. Plants of the third group make quick and active movements to capture the
prey. The quick closing mechanism of the metamorphosed leaves (trap) occurs by
the rapid change in the turgidity of the mesophyll cells, with ATP consumption.
Traps belonging to the plants of the genus Pinguicula and Byblis are functioning
due to sticky substances with a high content of carbohydrates, which are secreted by
specialized tentacular glands located on the leaves and which are directly used to
capture insects. Plants belonging to the genus Utricularia and Polypompholys live
in swamps. The pyriform vesicles which grow on the leaves can be closed with a
lid. Special glands will remove water from inside the vesicle, so that the lid will be
kept closed by the external water pressure. These plants secrete a substance with a
high content of carbohydrates that will both attract prey and help fixate the closed
lid. As soon as the victim touches the hairs of the lid the later opens and the
difference in pressure pushes the lid inwards while the victim is sucked together
with water. The lid closes, water is removed from the vesicle by glands and
digestion of the prey occurs by hydrolytic decomposition of complex organic
substances, in particular proteins, followed by their absorption.
absorbed and utilized by the rhizosphere microflora. Secretions from algae and
higher plants form a chemical environment that functions as an aquatic biocenosis.
Plants can eliminate final products of metabolism—this process is called
excretion, or substances that participate in metabolism, in which case it is con-
sidered secretion.
Thus, secretion is the active elimination of specific products of the metabolism
by first transforming active components of the cellular metabolism into less active
components. The process of secretion also requires the participation of active
mechanisms of substance transport, accompanied by consumption of metabolic
energy.
Excretion is the removal of remnants, waste and final products of metabolism,
unnecessary for further growth and development of living organisms and is per-
formed along the concentration gradient without the expense of metabolic energy
(ATP). A separate phenomenon is represented by water removal from plants
through various mechanisms—transpiration, guttation (Fig. 9.1) and plant tearing.
Eliminations are represented by plant proteolytic enzymes, nectar sugars, waxes
and terpenes, water and mineral salts (Fig. 9.2) and can be grouped into:
(1) internal eliminations that remain within plants;
(2) external eliminations which are excreted in the environment (Table 9.1).
In the 1970s, taking into account the ecological role assigned to these elimi-
nations for plant interaction within the biocenosis, the term “exometabolites” was
proposed for substances eliminated by plants under normal conditions and “path-
ological eliminations” or “stress metabolites” for substances that are secreted under
stress.
9.2 Excretion
Excretion is the removal of the surplus of substances from plants (waste, toxic
waste), whose accumulation would disturb homeostasis of the internal environment.
Excretion in plants differs greatly from that seen in the animal world. These fun-
damental differences are a direct consequence of the physiology and lifestyle of the
representatives of the two kingdoms—animal and vegetable. Plants are autotrophic
organisms, which are capable of synthesizing organic matter, forming small
252 9 Elimination of Substances in Plants
amounts of waste and therefore it was not necessary during development to develop
a well-differentiated excretory system. Representatives of the animal kingdom,
being heterotrophic and using organic substances in their nutrition, evolved to
eliminate nitrogen waste as urea, uric acid and ammonia. Whereas plants do not
feed on protein and don’t possess muscle activity (key factors of nitrogenous waste
formation), they don’t form this type of waste and small amounts of ammonia
formed in the metabolic processes are eliminated by diffusion.
Plants are primary producers and synthesize sufficient amounts of all organic
compounds required for growth and development. Only the amounts of substances
immediately required for the biological processes are synthesized at any moment in
plants, which explains why the plant body does not contain surpluses of protein,
respectively, does not eliminate nitrogenous waste. But if the proteins are decom-
posed to amino acids, they are reused in the biosynthesis of other proteins.
The three final products produced as a result of cellular metabolism in plants—
O2, CO2 and H2O—can be used again by plants as initial material for other reac-
tions. In the presence of sunlight, plants form as a result of photosynthesis much
larger quantities of molecular oxygen than are necessary for respiration. Thus, the
excess of O2 is released into the atmosphere by diffusion.
Many organic substances and metabolic residues are deposited in plant dead
tissues, in bark or in leaves that fall periodically. The substances destined for
excretion are removed also through sepals, petals, fruits and seeds, although at a
smaller extent.
Mineral salt ions absorbed by the plant root system are deposited together with
other cations in the form of insoluble crystals which stay in the cell, but are neutral
for their functional activity. For example, plants absorb both Ca2+ and SO42− ions,
but sulfates are included immediately in the process of amino acid (cysteine, cystine
and methionine) synthesis, while the divalent calcium cations keep accumulating.
Neutralization takes place due to interaction with the oxalacetic and pectic acids,
forming neutral substances insoluble in water such as calcium oxalate and pectate.
The later “cements” the medial lamella of plant cells. Lack or deficiency of Ca2+
ions leads to maceration of plant tissues. Other ions (Fe2+ and Mn2+) and some
organic acids (nicotinic acid, and tannins) are transported in leaves where they
accumulate, conferring a specific color before detaching.
In the majority of aquatic plants, most of the waste products pass directly into the
environment through diffusion.
Excretion of substances in higher plants differs according to the stage of vege-
tation, the capacity of excretion substantially increasing after flowering. Towards
the end of the vegetating season a series of substances is eliminated: previously
absorbed minerals, as well as organic substances such as acids, carbohydrates,
proteins with simple structure, vitamins, hormones, enzymes.
Ageing of protoplasmic colloids accompanied by a decrease in their ability to
retain ions, makes the precipitations from the end of the vegetating season to “wash”
plants, sequestering significant quantities of ions, minerals, substantially reducing
forage and fruit quality. For instance, wheat during maturity eliminates K+, Ca2+ and
Mg2+ ions in considerable amounts while vine releases K+ and Ca2+ ions.
9.2 Excretion 253
Fig. 9.3 Effect of root system exudates on nutrient availability (Dakor and Phillips 2002). AO
Organic acids; AA amino acids, including phytosiderophores; CF phenolic compounds
Around the plant roots an ecological environment is created that is different from
that of the surrounding soil and which is called rhizosphere, formed by plant
exudates that attract microorganisms through chemoattraction (Fig. 9.3). The
microflora of the rhizosphere, especially that of legumes, produces organic acids
(e.g. Ketoglutarate) used as solubilizers for phosphoric compounds and silicates. It
was shown that the wheat roots excrete glucose, flavones and nucleotides, while pea
plants eliminate only glucose and nucleotides. Through their roots, plants also
excrete other substances: amino acids, biologically active substances. Pea plants
eliminate much larger quantities of amino acids compared to oat. In general,
legumes often excrete biotin, niacin and pantothenic acid and less frequently
riboflavin and thiamine.
9.3 Secretion
Conquering the terrestrial environment by plants and increasing their body size
have dictated the development of the central conducting system in order to supply
the cells with water. It is precisely these phenomena that have contributed to the
appearance of lignin in cell walls (Fig. 9.5). Lignin has not been detected in current
and ancient aquatic plants; in the cell walls of a number of mosses there are
aromatic compounds, while true lignin appears only in ferns. In contemporary
higher plants, lignification represents the process of cell wall impregnation with
lignin, which is deposited between the cellulose microfibrils. In this manner lig-
nification occurs in hazelnut walls, nut mesocarps, wooden vessel walls etc. Lignins
are aromatic polymers contained in the tertiary cell walls and which confer them
their specific rigidity. Lignification is a process that occurs throughout the life cycle
of plant cells and increases at the end of the differentiation process. More intensely,
cell wall lignification occurs after completion of cell growth.
On the surface of the epidermis an outer skin called the cuticle forms (Fig. 9.6).
The cuticle consists of cutin and waxes and has a thickness of 1–15 nm, having a
low permeability for water and gas. The cuticle appeared in archaic terrestrial
plants, which resemble contemporary mosses and can be detected in sediments of
the silurian period. It is assumed that, with the advent of the cuticle, terrestrial
plants have adapted to water scarcity and survived under unfavorable conditions of
life. At the same time, the cuticle protects plants from short-wave radiation and has
implications on the rates of water and CO2 exchange.
The cuticle is formed by depositing cutin produced by the cytoplasm of epi-
dermal cells on the external cell walls and contains lipid polymers with 16 and 18
carbon atoms and minor amounts of phenolic compounds. The fatty acids of these
polymers contain two or more hydroxyl groups. Kolattuduy supported the idea that
cutin biosynthesis takes place on the outside of the cell wall, using monomers
formed in the smooth endoplasmic reticulum that are transported to the surface of
the cells in the form of vesicles or droplets. The cutin layer is coated with poly-
saccharide components of the cell wall (e.g. cellulose) and forms the cuticle, a
structure containing numerous molecular pores.
The cuticle surface in many plants has a polymer layer more hydrophobic than
cutin, consisting of waxes. Waxes deposited on the external surface of the cell walls
are a mixture of cerides: esters of fatty acids and hydroxyacids (with 15–35 carbon
atoms) with alcohols (having 28–34 carbon atoms). These substances also contain
free molecules of fatty acids, alcohols, ketones, hydrocarbons, aldehydes etc. The
waxes are synthesized by epidermal cells and transported outside through the
plasmodesmata and cuticular pores. Wax components are transported through the
cell wall embedded in a lipo- or glycoprotein “capsule”, which has a hydrophilic
outer surface and can, thus, be transported through the hydrophilic cell wall. It was
indicated that transport of the precursors of epicuticular waxes happens in a dis-
solved state. These substances pass through the ectodesmata and the cuticular pores
onto the surface of the cuticle where solvents evaporate and waxes crystallize. Wax
9.3 Secretion 257
Fig. 9.7 Longitudinal section through a salt secreting gland of Tamarix aphylla. The surface
transfusion can be seen on the left side of the main wall. Small vacuole proliferation was detected
near the secreting cells and their apparent association with the plasmalemma in the region of wall
extinction (Echeverria 2000)
deposition on the outer cell walls reduces water loss from tissues through the
process of transpiration. Each type of wax is deposited in a specific pattern (we can
distinguish 14 types)—from amorphous layers (the leaves of beet, the common
bean) to various forms of filaments and tubules.
Another process, suberization represents the deposition of suberin layers on the
inside of the epidermis and endoderm cell walls. This process also occurs in
mechanically damaged cells, contributing to the isolation of the tissues that were
attacked by parasitic organisms from the healthy ones.
Gelation represents the process of excessive production of pectic substances,
which in contact with water forms mucilage. This process is found frequently in
peach fruits.
Mineralization is the impregnation of the cell wall with silicon dioxide in
grasses, sedges, diatoms etc., with salts of calcium, carbonates and, less frequently,
sulfates in some woody plants. Trough mineralization of the cell walls the latter
acquire a greater mechanical strength.
Salt-secreting glands are located in the epidermis of the leafs or stems and have a
similar structure to that of hydathodes. These glands are characteristic for some
species of halophytes and are designed to collect and remove excessive amount of
mineral salts (Fig. 9.7). Such secreting glands are found in statice, which actively
eliminates chlorine and passively eliminates sodium.
Nectary glands are made of a special type of epidermal cells and are localized on
some floral or even extrafloral components on stipels, receptacles, stamen filaments,
on the ovary or on the style (stilus) (Fig. 9.8).
258 9 Elimination of Substances in Plants
Fig. 9.8 Nectariferous glands in Dionea muscipula (a), Drosera intermedia (b) and Phyostegia
virginiana (c)
Fig. 9.9 The nectary gland in Croton glandulosus. Nectar secretion may cause the separation of
the cuticle from the outer periclinal walls of palisade cells that form the secretory epidermis
Nectariferous glands (nectaries) (Fig. 9.9) secrete substances used in pollen ger-
mination and fecundation of the ovum. Unused nectar can be absorbed by the ovary
for additional nutrition of the generative parts of the plant. Intrafloral nectaries have
the role to attract insects that carry out cross-pollination, while the extrafloral
nectaries serve to remove the excess of glucose in the periods of high photosyn-
thetic activity. The nectar has phytoncide and bactericidal properties, protecting the
ovary from various microorganisms. Extrafloral nectaries defend the plants from the
insects feed on them.
There are two possible ways of transporting the nectar in the cells of the nectary
glands. The transport of pre-nectar from cells to the nectaries can occur by passive
transport through the apoplast due to the proton pump localized in parenchymal
cells of the passage elements, which eliminates the pre-nectar from the passage
elements under pressure. Pre-nectar moves from cell to cell and through the sym-
plast, the plasmodesmata, turning into a type of nectar specific for the plant species
in the cytoplasm of the nectary gland cells, followed by its elimination. This
transport is carried with consumption of metabolic energy, as confirmed by
increasing oxygen intake during secretion and an increase in the number of mito-
chondria in the cells of the nectaries. Both the symplastic and apoplastic transport
9.3 Secretion 259
can be identified at different stages of the traveled path. At the same time, in nectary
glands, with lignified and suberized cell walls, in which the transport through the
apoplast is excluded, the transport happens exclusively through the symplast.
Elimination of the nectar from nectariferous epithelium occurs in the form of
droplets and is similar to guttation.
The morphology of the structures that eliminate terpenes are of a great diversity—
solitary cells or groups of secretory cells. Isolated secretory cells called idioblasts
can be found in different plant tissues: leaves, stems, etc. Glandular hairs represent
outgrowths consisting of one or more cells derived from the epidermis and,
sometimes, from the hypodermis. They are located on the leaves and stems of the
plants and have a terminal part formed of one or more secretory cells. Glandular
hairs from Primula, Alnus and Populus producing flavonoids are characterized by
the presence in the terminal cell of a large number of ducts of the endoplasmic
reticulum, while the hairs producing carbohydrates are characterized by well-
developed Golgi complexes. The essential oils synthesized by Mentha piperita
accumulate in the vacuoles of the secretory hairs, while stinging hairs of Urtica
dioica accumulate histamines and acetylcholine. The secretions are released
through pinocytosys in the space between the plasmalemma and the cell wall, from
where it diffuses towards the exterior, accumulating in the subcuticular space.
Glandular papillae are composed of epidermal cells, elongated outwards. They can
be found in species of the genus Rosa, Syringa, Lilium, Hyacinthus and the
secretions consist of volatile oils.
The osmophores are specialized glandular cells which secrete essential oils that
determine floral aroma in a group of plants. Various parts of the flower can morph
into osmophores accepting a hair-like, wing-like form etc.
From the essential oils almost 1,000 organic components are extracted—terp-
enes, carbohydrates, aldehydes, alcohols, ketones, phenols and others. The linalool
is characteristic for oils derived from the lily of the valley, oranges, coriander,
eucalyptus, roses, geraniums. Several phytohormones also belong to the class of
terpenes—the abscisic acid (C15H20O4), gibberellins (C19–20), steroids (C28–30). The
composition of essential oils specific for each plant species is characterized by the
presence of tens of terpenoids (Fig. 9.10).
Lactiferous cells consist of long secreting cells, branched or unbranched, arranged
in rows (unbranched lactiferous cells) or made of cells placed end to end, which have
their separation walls reabsorbed (articulated lactiferous cells). During the growth of
the lactiferous cells, a single large vacuole is formed for the accumulation of latex—
an emulsion containing 50–80 % water, as well a series of substances in the form of a
suspension: carbohydrates, tannins, glycosides, calcium malate, essential oils,
waxes, gums, resins, aromatic acids, polyisoprenic carbohydrates and sometimes
toxic alkaloids. The color of latex is white, in contact with air, it strengthens.
260 9 Elimination of Substances in Plants
Fig. 9.10 Terpenophenols in hops (Humulus lupulus). The paternal genotype produces cones
(a) used in beer fermentation. Glandular trichomes (lupulin glands), covering the cone bracts
(b) represent the main place of terpenophenol biosynthesis in hops
Among higher plants more than 400 species of carnivorous species are known that
have various mechanisms of hunting and different secretory glands. These plants, in
addition to autotrophic nutrition processes, are able to use small animals (insects,
juvenile fish) as a source of food.
The leaves of insectivorous plants turn into special traps, capable both to pho-
tosynthesize and capture prey. Usually, the latter sticks to the leaves, whose glands
eliminate a sticky mucus (as is the case of sundew—Drosera rotundifolia) or falls
into traps in the form of jugs, tubes, bags that are brightly colored and eliminate
sweet, aromatic secretions (Sarracenia, Heliamphora).
The principle of the trap is based on catching the victim with the help of quickly
closing leaves or by sucking the prey with the help of a sudden water current,
created with the help of a vacuum formed in the trap. Common to all the types of
traps is the need to attract insects with nectar and polysaccharides (in the mucus),
which are either secreted by the specialized trapping organ or by glands localized
near the trap. Responsiveness of the trap mechanisms is triggered by the irritation of
sensitive hairs that is caused by prey movement (Figs. 9.11 and 9.12).
Secretions produced by digestive glands contain proteolytic enzymes and
mucilaginous, sticky substances, ions of sodium, potassium, magnesium, calcium
and chloride in a concentration of 20 mmol. Secretions also contain amylases,
glucuronases and cell wall hydrolytic enzymes, which are produced by the endo-
plasmic reticulum and ribonucleases, proteases, phosphatases generated by the
Golgi complex. Insects stick to the polysaccharide containing mucus, which fixes
them in place.
The biological importance of the secretory processes in insectivorous plants
consists in achieving heterotrophic digestion, in addition to the process of
Fig. 9.11 Drosera intermedia. General aspect (left) and nectary glands (right)
262 9 Elimination of Substances in Plants
stability of the water content. The water regime of plants is one of the determining
factors for a normal metabolism of the organism, hence its role in homeostasis
(Table 9.2).
The main processes by which the plants are eliminating the water are transpi-
ration, tearing and guttation (Fig. 9.13).
Removing water in the form of vapors represents transpiration—the process
that is characteristic for all terrestrial plants and which occurs with varying intensity
in various plant organs (except for the roots) and in different groups of plants
(Table 9.3).
The process of transpiration does not require metabolic energy, it uses light
energy that by being absorbed is converted into heat which fuels the process. From
this point of view, the mechanism is called passive. But in terms of the suction force
it generates it exceeds the active mechanisms of absorption and movement of water
through the plant, which is carried by the root system, based on root pressure.
Removing water from the plant is essential because it ensures: crude sap ascension
264 9 Elimination of Substances in Plants
Table 9.3 Intensity of stomatal and cuticular transpiration in different groups of plants
Group of mg H2O (dm2/h)
plants Stomatal Cuticular % from stomatal
transpiration transpiration transpiration
Hydrophytes 1,800–1,400 – 60–70
Helyophytes 1,700–2,500 200–300 up to 20
Ombrophytes 500–1,000 50–250 up to 25
Pomoideae 700–1,000 120–160 10–20
Prunoideae 400–700 80–110 15–20
Vitaceae 400–500 80–90 17–24
Cactaceae 200–600 3–10 1–2
through the xylem vessels, avoidance of cell saturation with water and plant
overheating (vaporization of 1 g of water is accompanied by absorption of 2.2 kJ of
energy). Due to these reasons, transpiration is considered a necessary physiological
evil for plants.
Transpiration occurs through stomata and lenticels but also through the external
protective layers: cuticle, suber, etc.
Osteolar transpiration is correlated with the rate of water provision and their
degree of openness. Stomata have a wide distribution on the leaf surface, depending
on the species. Approximately 71 % of the woody species are hypostomatic and
almost all of the crop plants are amphystomatic.
Water from the xylemic vessels is transported through the apoplast into the
lacunar parenchyma cells and the process of evaporation takes place in the sub-
stomatal cavity. Water vapor transport through the lacunar spaces, has a lower
extent. Air from the substomatal cavity is saturated with water vapor, while the
ambient air has a low relative humidity (40–60 %). As a result, water vapor diffuses
into the environment from the substomatal cavity. After harvesting vegetables, the
stomata close in about 10 min which helps to maintain turgidity during further
processing.
Cuticular transpiration has a reduced extent due to the low permeability of the
lipid components of the cuticle. This type of transpiration makes up 1–2 % of the
total transpiration in Cactaceae plants, between 10 and 25 % in mesophilic plants
and reaches up to 60–70 % of the total transpiration in hydrophytes. In plants with
very thick cuticle, like the Hedera helix, cuticular transpiration does not occur at all.
Lenticular transpiration is evident in stems, fruits and leaves of conifers, con-
taining lenticels which are gaps in the epidermis, periderm or modified stomata. By
means of the lenticels, living cortical cells are connected to the ambient atmosphere.
When provided with abundant amounts of water and in the presence of a high
content of water in the atmosphere, several plants, especially young ones, eliminate
the raw sap through the upper leaves in the form of droplets. This phenomenon is
called guttation. In dicotyledonous plants, guttation occurs through special open-
ings called hydathodes, localized on the tips of the leaves or the lobes and can be
9.5 Water Elimination in Plants 265
noticed at night or in the morning, during spring and summer. Among cultivated
plants, considerable levels of guttation can be noticed in potatoes, cereals, primula.
Some tropical plants such as Caesalpinia gilliesii have an abundant guttation such
that under the tree water is dripping.
Guttation was observed in more than 300 genera of plants and it is characteristic
for tropical and equatorial plants. Like in the case of plant tearing guttation is an
eloquent proof of the root pressure.
At the basis of this phenomenon lies the disparity that is created between water
absorption by the roots and its elimination in the form of vapors. In grasses,
guttation is observed at a young age—elimination of water takes place not through
the hydathodes, but trough the tips of the leaves, after the layers of young cells
rupture under the internal pressure. In the guttation water of the cereals, along with
other substances (salts of calcium, potassium, phosphorus), malic acid is almost
always present. Guttation in this group of plants can be easily induced by covering
the pot in which the plant (wheat, barley and maize) grows with a glass bell for a
certain period of time. From the biological point of view, guttation can be regarded
as an adaptation of the plant in order to remove the water excess from the body.
Due to this phenomenon, the equilibrium between the absorbed and eliminated
water is maintained.
If plants couldn’t remove water excess, the later would penetrate the intercellular
spaces and would cause tissue death by suffocation. Some physiologists believe
guttation is important because, along with water, plants eliminate many toxic salts.
Besides guttation the phenomenon of plant tearing can be noticed which is
happening due to the root pressure demonstrating an active, purely physiological
water absorption and conductance mechanism. The root pressure is the force that
pumps water into the symplast of the endoderm through the conducting vessels of
the central cylinder. During the spring, when many plants don’t have leaves, the
only force that moves the water through the plants is the root pressure. The value of
the generated force depends on the accumulation of plastic substances in the cor-
tical parenchyma cells and on the energy metabolism of the cells. In some cir-
cumstances, depending on the root pressure values, the crude sap can elevate to a
certain height of the plant. This effect can be easily noticed in vines during early
spring, which when cut releases large amounts of sap. This phenomenon was called
“tearing”. It can be observed in many plants also during the summer, when the
aerial part of the plant is cut. Moreover, crude sap can be collected, and its chemical
composition–determined. Also the value of the root pressure and the osmotic
pressure of the sap can be measured. The value of the root pressure is relatively low.
In some trees and vines values of 2–3 atm, where detected which can lift water up to
15–20 m high. Once the leaves come into action the leaf suction force is generating
a counterforce in the conducting vessels and often prevent sap eliminations after
mechanical lesions. In perennial plants, this phenomenon manifests itself especially
during the spring (birch juice) while in herbaceous species—throughout the veg-
etation period. The crude sap eliminated by the process of tearing contains mineral
salts, organic substances, sugars, etc. Summer raw sap contains ash—derived ele-
ments, organic acids, but no sugar.
266 9 Elimination of Substances in Plants
Plant tearing lasts from a few days up to 5–6 months, and the eliminated water
amount varies from one species to another. A palm tree can eliminate by tearing
about 10 L of sap, a birch tree—5 L, and a vine plant—almost 1 L, with a force of
1.6 atm, elevating water to about 16–21 m.
The crude sap elimination mechanism was explained by A. Sabinin by the
different nature of the metabolism occurring in the cells surrounding the xylem
vessels, which leads to the creation of a difference in the suction force, as result of
which, at one end of the cell water is being absorbed while at the other—eliminated.
If a plant is exposed in a solution with the osmotic pressure equal to the osmotic
pressure of the raw sap, tearing ceases immediately. Tearing is closely related to the
vital activity of the plant. If soil aeration is low, the rate of water elimination by
tearing is reduced significantly.
• After elimination of waxes from the surface of the plants, their permeability for
water increase over 500 times. The osteoles of many angiosperms and gym-
nosperms are also coated with wax containing structures that are designed to
minimize water loss from the aerial niches of the osteoles. The waxy pellicle on
the osteole in some xerophytes intensifies, during the night, water condensation
from the air.
• The suberin is formed during secondary growth, causing Kaspari thickenings in
endodermal cells or impregnates the outer cells of the potato tuber, during the
mature stage.
• The leaves of Laurus nobilis contain large cells that secrete volatile oils. In the
bark of Quercus species and in the medular tissue of Rosa species cells can be
found in which substances like tannins, alkaloids and heterosides are synthe-
sized and accumulated.
• Plants from the Tiliaceae family have cells containing mucilages derived from
cytoplasm degeneration or from the cell wall gelling.
• In the petiole of Begonia, Prunus, Allium species cells that accumulate crystals
of calcium oxalate can be found.
• In cells of Liriodendron and Magnolia volatile oils are accumulating in vesicles
formed by invagination of the plasmalemma. The accumulation of these sub-
stances takes place in cells of the leaf mesophyll, of the fruit mesocarp and, in
some cases, in roots and stems.
• Secretory glands in plants of the Fabaceae family are rounded, consisting of
6–12 cells located radially, while in species of the Compozitae family they are
elongated, bicellular.
• The latex of the Papaver species contains alkaloids, that of the Ficus—protein
substances, that of the Musa—tannins while Carica papaya contains papain.
• On a 1 year old branch of Gledistria triacanthosa with a length of 20 cm
72–210 lenticels can be found, while tubers of Solanum tuberosum have a
lenticel per 1 cm2.
• A plant of Zea mays absorbs during the vegetating season about 250 L of water,
of which about 98 % is eliminated by transpiration.
Glossary
Biocoenosis The term designating a living community including also the atmo-
sphere and the soil. The biocoenosis comprises the phytocoenosis, zoocenosis
and microbiocoenosis.
Halophytes Plants that can grow on soils with high salt content due to the presence
of a number of characters and traits that have emerged under the action of
specific environmental conditions and natural selection during evolution.
Homeostasis An autoregulatory process by means of which the organism main-
tains a certain chemical composition a set of physical and morphological
parameters and can re-establish a disrupted balance, either by returning to its
original state, either by returning to an equivalent state.
Median lamella The cellular structure that appears in the telophase of the mitotic
process between two daughter cells and is composed of insoluble pectic sub-
stances (protopectin), which serve as a ligand between cells. Tissue softening in
fruits is caused by the enzymatic decomposition of pectic substances from the
middle lamella resulting in easily separating cells, a process which characterizes
the stages of maturity and senescence.
Terpenes Organic substances widely spread in plants as constituents of essential
oils, also some insects have terpenes as a component of protective substances
and pheromones. Sometimes the term designates only the corresponding car-
bohydrates while their oxidized derivatives are called monoterpenes.
References
organism. Plants try to prevent toxic gases from entering the leaves or to integrate
them in the metabolism, to immobilize the ions of heavy metals in the cell wall
reducing their toxic effect.
Historical Background
1896—B. Lindforss demonstrates the role of carbohydrates as cryoprotectants in
freezing cells.
1904—B.D. Zalenski studies the role of the anatomy of leaves in drought
resistance.
1931—I.I. Tumanov launches the idea of plant hardening in frost.
1958—Rudolf W. and H. Kapert underline the role of unfavorable factors in the
development of functional and structural disorders in plants.
1962—B.P. Stroganov demonstrates that plant adaptation to different kinds of
salinization is different.
1962—F. Ritossa reveals heat shock proteins.
1971—M.N. Christiansen finds that low temperatures cause metabolic aberrations
in plants.
10 Physiology of Plant Resistance to Unfavorable Environmental Factors 273
Brief Updates
A large group of proteins that are involved in stress responses are heat shock
proteins (HSP) or chaperons, which are synthesized in cells following exposure to
high temperatures. Such proteins are found in microorganisms, animals and plants,
presenting high structural similarity, which is indicative of their evolutionary
conservation. For example, the structure of the HSP with the Mr 70 kDa is similar
in insects, birds, mammals, fungi and plants. In plants, HSPs can have a big mass
(Mr between 60 and 110 kDa) and a small mass (with Mr of 15–35 kDa). Their
function is to bind normal proteins and prevent their aggregation induced by high
temperatures, the energy process is ATP-dependent.
HSP synthesis is initiated when temperature changes with 8–10 °C compared to
the normal. In such stressful conditions, the genetic program stops normal protein
synthesis and replaces them with stress proteins. Synthesis of mRNA encoding HSP
starts already after 5 min and the maximum is recorded after 2–4 h and followed by
decrease. It has been demonstrated that HSP are synthesized also during other stress
situations, such as, for example, the action of heavy metals.
Induction of HSP synthesis serves as a model for the study of the molecular
mechanisms regulating gene expression in the process of adaptation to stress fac-
tors. In the future it is expected to obtain transgenic plants containing genes
encoding HSP, which would contribute to an increased resistance to various
extreme conditions.
Dehydrins are proteins similar to the HSPs that are synthesized in the late stages
of embryogenesis under the influence of abscisic acid (ABA). Their elevated
content correlates with cell dehydration that occurs during this developmental stage.
In cotton there were found 18 molecules of mRNA, 13 of which can be induced by
exogenous treatment with ABA. It is assumed, therefore, that these proteins may be
involved in the protection of plants during water deficit. The expression of the
genes encoding such proteins may be induced by ABA, by low temperatures, by
drought and salinity.
Fig. 10.3 The main types of adaption of living organisms to the environmental conditions
276 10 Physiology of Plant Resistance to Unfavorable Environmental Factors
During winter, plants, especially perennial and multi-annual grasses are influenced
by unfavorable conditions caused by low temperatures. In addition, perennial plants
may suffer from other damaging phenomena such as asphyxiation under snow (e.g.
winter wheat may perish due to the thick snow layer), asphyxiation under water,
uprooting, alternating low and high temperatures and winter drought.
Asphyxiation under snow represents the death of plants trapped under a thick
layer of snow for 2–3 months under moderate winter conditions at a temperature of
about 0 °C in unfrozen ground. Snow is a bad conductor of heat, so the temperature
rises under the snow sheath to about 0 °C, causing an intensification of the respi-
ratory process. Plants start utilizing carbohydrate reserves, whose amount decrease
from 20 to 2–4 % and plants may die due to exhaustion of energy resources, a
phenomenon called inanition.
It was noticed that in thermophilic plants, a decrease in temperature is accom-
panied by a sharp increase in cytoplasm viscosity and the oxidation-phosphorylation
correlation is disturbed. Thus, plants lose resistance to adverse conditions and
10.2 Unfavorable Factors of the Winter-Spring Period 277
during spring are easily attacked by Fusarium nivale and perish. Consequently, in
areas with heavy snow crop varieties containing increased amounts of carbohydrates
are needed.
Plants often perish during the winter due to the formation of a thin crust of ice on
the surface of the soil, which causes plant uprooting. Uprooting is the process
through which the ice crust detaches the top layer of the soil together with the plant,
leading to rupture of the root and organism death. Uprooting is conditioned by the
attraction of water from the deeper layers of the soil to the ice layer formed on the
surface. If, as a result of the frost, the layer at 2.5–5.0 cm from the soil surface
freezes and if there is no further cooling, this layer begins to absorb water from
deeper soil layers. As a result, the soil crust forms a frozen ice crust which elevates
and detaches the frozen soil layer from the unfrozen one damaging plant roots.
Frost resistant varieties which are characterized by a greater extensibility of the
roots support these conditions easier, while those with poorly developed root
systems perish. In order to prevent this phenomenon, it is necessary to create
varieties with high extensibility of the root system and to use methods which reduce
the amount of water on the surface of the soil.
Another common phenomenon in certain regions of the globe are spring frosts.
During the spring time plants die from an alternation of high and low temperatures.
Thus, plants that survived the winter at −30 °C can die in the spring at less extreme
temperatures, because they have completed the processes of hardening and began
the processes of growth.
In early spring, water accumulation from melting snow can cause plant
asphyxiation under water. Typically, these phenomena occur in the spring, but can
also occur during the winter when temperatures stay for a long time around 0 °C
and plants remain for a long period under water. Under excessive humidity con-
ditions, in these plants due to oxygen shortage, aerobic respiration decreases in
intensity while anaerobic respiration intensifies.
Accumulation of toxic substances (alcohol) as a result of anaerobic respiration
leads to plant poisoning. In addition, excessive consumption of organic substances,
weakens plant fitness (Fig. 10.4).
Fig. 10.4 Influence of oxygen deficiency on various metabolic processes (Igamberdiev et al.
2004)
temperature the water regime of the plants is disturbed. The main cause of the
harmful action of low temperatures on heat-loving plants is the perturbation of the
functional activity of the membranes, as a result of the transition of the saturated
fatty acids from the liquid crystalline state to the gel state. This alters the normal
balance in substance exchange, while a longer action of this negative factor can lead
to death (Fig. 10.5).
Agricultural plant resistance to cold can be increased by hardening seeds with
low temperatures (with the periodicity of 12 h—1–5 °C, 10–20 °C). The same
10.3 Plant Resistance to Cold and Frost 279
Fig. 10.6 Acclimatization to frost in different Arabidopsis lines. a Three week plants were
acclimatized at 1 °C with a photoperiod of 16 h for 0 (NA), 1, 2 or 3 days or acclimatized for
3 days followed by deacclimatization at 21 °C for 1 day. These plants were frozen at −4 °C (left),
−7 °C (center) or −10 °C (right); b three week plants were not acclimatized (NA) or kept for
3 days at 1 °C in darkness or in conditions of a 16 h photoperiod; c Three week plants were
acclimated to a 12 h photoperiod at a high temperature (1 °C) during the day and a low temperature
(−1 °C) during the night (1 °L/−1 °D), at a low temperature during the day and a high temperature
during the night (−1 °L/1 °D), or at 1 °C (1 °L/1 °D) for 2, 3 or 5 days before freezing at −8 °C;
d three week plants were acclimatizated at 1 °C and 16 h of daily light for 2 or 3 days in the
presence or absence of 50 mM DCMU before freezing at −8 °C
method is also used for treating seedlings, except that in this case a 25 % solution of
microelements or ammonium nitrate is added for 20 h.
The frost resistance of plants is their ability to survive under the influence of low
temperatures well below 0 °C (Fig. 10.6).
It was established that freezing temperatures below −20 °C are specific to about
42 % of the globe.
Low temperatures have different effects on plants. The main causes of cell death
at low temperatures are cell dehydration and mechanical trauma caused by ice
crystal formation.
A rapid decrease in temperature in extreme conditions leads to intracellular ice
formation and cell death.
A gradual decrease in temperature, which is more likely to happen in natural
conditions, leads to ice formation in intercellular spaces. The formed ice crystals
remove the air and frozen tissues may seem transparent.
Specific physiological adaptations allow plants to partially avoid wounds that
occur after frost. It is known that in case of tissue freezing the biggest damage is
produced by the crystals of ice formed within the cell. These crystals grow, break
cell membranes and the cell dies. Increasing the concentration of dissolved sub-
stances in the vacuolar sap, mainly carbohydrates, protects plants from frost
damage, because it increases the probability of formation of large ice crystals.
Another method of protecting plants from damage caused by frost is by
increasing the quantities of proteins, which are able to recruit water into their
280 10 Physiology of Plant Resistance to Unfavorable Environmental Factors
hydration layers. Hydration water practically does not freeze. It is held around
protein molecules with larger forces which prevents formation of ice crystals.
Therefore, the more proteins of this type are synthesized in a cell, the more resistant
is the cell to frost.
It was found that many plants synthesize such proteins in autumn while in spring
they are utilized in metabolism.
Fig. 10.8 Plant organism response to water deficit (Bohnert et al. 1995). Under the influence of
drought various cellular processes can be affected. These changes allow the plant to maintain
metabolism and restore the conditions that allow continued growth under stress
seeds, small fruits etc. Long-term drought causes cytorrhysis (strong dehydration of
the cytoplasm) which destroys the integrity of the cell.
For many years it was thought that water is available to plants, as long as, the
soil moisture does not reach the wilting coefficient beyond which water remains
inaccessible. According to this conception, physiological processes, plant growth
and development proceed normally until reaching the wilting coefficient of the soil.
But recent scientific research has shown that the exchange of substances is influ-
enced even by a small shortage of water. Such a deficit occurs inside tissues and
cells long before soil moisture reaches the wilting coefficient. Water scarcity in
plants acts on a range of physiological and biochemical processes such as water
uptake, root pressure, seed germination, stomata movements, transpiration, photo-
synthesis, respiration, general enzymatic activity, etc.
Plants that had a strong water deficit even for a short period of time do not return
to a normal exchange of substances. The water balance and the degree of affection
depend on a number of internal and external factors related to the type of plant
(drought resistance, the depth of the root system penetration into the soil and its
branching, the stage of development), the number of plants per unit of area, weather
conditions (temperature and air humidity, wind, light, amount of atmospheric
284 10 Physiology of Plant Resistance to Unfavorable Environmental Factors
Fig. 10.10 Northern-blot analysis of transgenic tomato plants (Tsai-Hung et al. 2002) Total RNA
(10 µg) was extracted from control plants (M, track 1) and from T1 transgenic plants which are
characterized by an exaggerated expression of CBF1 (C repeat/dehydration-responsive element
binding factor 1) (tracks 2–9). The used cDNA samples (Arabidopsis CBF1, the GUS reporter
gene from pCAMBIA 2301, CAT1 from tomato and β-TUBULIN, rRNA) were labeled with P32
precipitation), soil-related factors (the quantity of water in the soil, the osmotic
pressure of the soil solution, the physical properties and composition of the soil).
The influence of the water deficit on metabolic processes depends largely on the
duration of drought action. Thus, in cereals a long withering period increases the
rate of RNA and protein degradation, which is leading to an increased amount of
nitrogen-containing non-protein organic compounds and their deposition in the
stem and the ear. As a result, during drought there is an increase in the quantity of
protein in seeds and a decrease of their content in leaves.
Water scarcity also affects substantially the exchange of carbon, lowering the
intensity of photosynthesis and reducing the amount of synthesized ATP during
photophosphorylation. Under the action of pedological and atmospheric drought,
the processes of translocation of photosynthesis products in other organs are slowed
down.
Drought, be it long or short, ultimately leads to lower plant productivity. In the
process of evolution, a group of plants has formed with a number of adaptations to
drought represented by false xerophytes (ephemeral and ephemeroids). To combat
the effects of drought it is required to plant forest belts in dry areas, to create
resistant varieties, able to adapt to drought conditions and to practice irrigation. In
order to increase plant resistance to drought, the method proposed by P.A. Henkel is
used, which consists in drying the swollen seeds due to which the emerging plants
will acquire xeromorphic qualities.
An important role in increasing plant resistance to drought, as shown by
K.A. Timireazev, is played by mineral fertilizers. Administration of phosphorus
during sowing, of nitrogen during tillering, and of K, B, and Cu during the critical
period significantly increases plant resistance to water scarcity. Under the influence
10.4 Plant Resistance to Drought 285
of B, Mn, Cu, Co, Mo, Zn the hydrophilic protein colloid content increases, as well
as their hydration degree, the viscosity of the protoplasm becomes higher and the
amount of “bound water”. It was demonstrated that Zn and B may favor the
formation of organic acids, which help fixate the ammonia produced during
drought, converting it into a harmless form. Microelements contribute to the for-
mation of phosphorylated compounds, which is of great importance, because the
phosphorylation processes in plants slow down during drought.
Plant growth depends largely on the amount of water available for the plants in the
soil. Approximately one third of the land area is occupied by regions with essential
moisture deficit, half of which, or about 12 % of Earth’s land is very dry. Regions
with moisture excess occupy about 9 % of the surface of the land.
If the amount of annual rainfall is greater than the amount of evaporated water
then the area is a humid area and if intense water evaporation takes place and the
rainfall is low, the area is called arid. Many agricultural regions are located in arid
zones and farming here is possible only by practicing artificial irrigation.
At the moment there is a system of measures to combat the drought, one of the
most important being artificial irrigation, which involves also administration of
mineral elements in different climatic zones.
However, for an efficient use of water resources it is important to know the
physiological parameters of the crop species that characterize their water regime.
One of the problems of irrigation is to determine the maximum and minimum
humidity allowable levels. The upper limit of allowable humidity is called soil
humidity. Exceeding the allowable upper limit of soil humidity is inadmissible
because excess water is not useful for plants, but on the contrary, is harmful because
it creates a lack of oxygen.
As the soil dries up until it reaches the humidity at which permanent wilting
occurs plants suffer increasingly from water scarcity. The humidity of constant
wilting cannot be used as an index for starting irrigation, as permanent wilting leads
to a water deficit in tissues and therefore decreases the productivity of plants.
Determination of the optimum levels of soil moisture, which do not cause physi-
ological disorders—the upper limit of optimum humidity, is important for con-
trolling the processes of plant growth and development.
In practical terms, however, it is very difficult to know these parameters. For this
purpose, various methods for their determination have been developed starting with
measurements of leaf saturation with water, of leaf osmotic pressure, of stomata
opening, of the generated suction force.
Nowadays, for different crops the values are known for the critical suction force
of the leaf, which correlates with the state of the plant that still does not cause crop
yield reduction. Identification and assessment of physiological indices for deter-
mining the forms and limits of more accurate and economical irrigation is an
286 10 Physiology of Plant Resistance to Unfavorable Environmental Factors
Currently, about 20 % of the cultivated areas and about half of the world’s arable
land are marked by salinization. According to data published by the International
Society for Soil Science, saline soils occupy an area of 3 million km2 only in
Europe and Australia.
Soils characterized by an excessive concentration of salt, may adversely affect or
even destroy the flora. In addition, improper irrigation is leading to salinization of
the soils. A harmful effect characterized by high concentrations of salt occurs after
the application of excessive doses of mineral fertilizers. Salts accumulate in the
ground, as in dry regions the soil is not washed enough by water to carry the salts in
the deeper layers of the ground. On the other hand, salts can be brought to the
surface by the upward flow of water in the process of evaporation. These soils have
a low fertility potential and the salt excess has many negative effects, including:
• increases the osmotic pressure of the soil solution making water absorption more
difficult;
• inhibits imbibition and germination of the seeds;
• inhibits the growth of the root system;
• has a toxic influence on the protoplasm;
• inhibits anabolic processes—photosynthesis and protein synthesis;
• prevents starch formation in somatic cells;
• causes oversaturation of the root system cells with salts, which leads to loss of
selectivity in the process of ion absorption, the later entering passively into the
cell in order to be accumulated in abundance in the vacuole.
In 1898 it was developed a theory according to which the action of salts on
plants consists in creating a “physiological drought” because the osmotic pressure
in plant cells is 15–20 atm compared to 30–50 atm in the soil. Plants can’t absorb
water, so these soils are called dry.
Salinization can be determined by the presence of: sulphate (Na2SO4), chloride
(NaCl), carbonate (Na2CO3) or it can be combined (Fig. 10.11).
A high saline content in the soil leads to the perturbation of the metabolic
processes. For example, it has shown that, under the action of salt excess, in plants,
the exchange of nitrogen is disturbed, leading to the accumulation of ammonia and
other toxic products.
The most harmful action on plants is played by the excess of carbonates as these
degrade in the soil forming strong bases (NaOH). An abundance of sulphates in the
soil leads to the accumulation of sulfur oxidation products that can be toxic to
plants. A high concentration of salts, in particular chlorides, can disrupt the process
of oxidation and phosphorylation i.e. plant supply with energy.
10.5 Plant Resistance to Saltiness 287
Fig. 10.12 ST overexpression (salinity resistance gene) increases resistance to salinity. The
picture shows germination of the control and transgenic ST genotypes. Four-day seedlings were
transferred to a fresh medium supplemented with 0, 50 and 100 mM NaCl and grown for 10 days
(a). Seedling root growth in the basic medium supplemented with NaCl has been established in
control samples and transgenic ST samples after 10 days (b). The white bar control samples; black
bar transgenic ST plants
from physiological drought. Under these conditions the absorption of ions from the
external environment is regulated by active transport mechanisms localized in the
cell membranes, but the control is relative. Thus, if in the external environment
toxic ions are present, they will be partially absorbed. It was established that with an
increasing soil salinity the absorption of Na+ ions and Cl− increased and of K+ and
Ca2+ decreased.
Increasing the osmotic pressure due to the accumulation of ions through
pinocytosis. An adaptive process in plants is the formation of pinocytotic vesicles
in root and cotyledonary cells of the halophytes facilitating an increase in the
osmotic pressure. It was proved experimentally in barley seedlings that, under the
influence of NaCl, pinocitotic vesicles appear that stimulate Na+ accumulation and
K+ efflux, increasing the Na/K ratio 100 times, while the Na/K ratio in halophytes
was 5 times higher. By invagination of the plasmalemma and vesicle formation in
the cytoplasm the compartmentalization of the high salt concentrations occurs, thus
protecting the enzymes from toxicity.
Increasing the resistance of the enzymatic system. Homeostasis maintenance
in halophytes is determined by increasing the resistance of the enzymatic system
that preserves its high activity under the presence of relatively high concentration of
toxic salts in the cell. It was found that enzymes isolated from halophytic plants
slow their work under the action of concentrated salt solutions, while enzymes
isolated from glycophytic plants completely lose enzymatic activity. Thus, ATP-
ases and malate dehydrogenases of halophyte species under high salt conditions had
higher levels of activity and varieties of the haloresistant Pennisetum americanum
had an elevated activity of their NAD-nitrate reductase, NADP-glutamate dehy-
drogenase, of the enzymes of glutamine and glutamate synthetase in comparison
with the same enzymes from salt sensitive species. (The high enzymatic activity of
malate dehydrogenase at high concentrations of NaCl reflects some structural
adaptations of the enzyme molecule.)
Increasing succulence represents a way to adapt to salinity. It was established
that succulent halophytes have high selectivity for chlorine absorption, which
accumulates in the intercellular space (30–35 %). Experimental work has shown
that NaCl slows down pectinesterase activity in Aster tripolium and salt is retained
by methylated pectic substances, increasing the plasticity of the cell wall and
facilitating cell elongation.
Increasing the osmotic pressure by accumulating organic substances.
Halophytes for which the accumulation of salts is toxic and whose roots have a low
permeability for salts adapted to high salinity by increasing the concentration of the
vacuolar sap, accumulating active osmotic organic compounds—carbohydrates and
organic acids, organic nitrogen in the form of glycine, beatin and proline which are
involved in osmotic regulation and act as protective substances.
Modification of protein biosynthesis. Under the influence of high salt con-
centrations protein synthesis is altered and accumulation of high molecular weight
proteins is enhanced as a measure of adaptation. The content of alkaline soluble
proteins and water increases in plants under high salinity conditions, which plays a
protective role. Metabolism rates decrease.
10.6 Regulation of Physiological Processes in Halophyte Plants 291
shown that the genes encoding vacuolar Na+/H+ antiporters increased salinity
resistance of Arabidopsis plants, cabbage, tomato, rice. Currently investigations are
carried out in order to obtain transgenic wheat plants resistant to salinity by
transferring the Na+/H+ gene to the vacuolar antiporter HNHX cloned from the
barley genome.
Currently the global public attention is drawn to the rate of ecological situation
worsening. Various issues such as continuing degradation of the environment,
reduction of plant and animal species number, decreased soil fertility, increased
content of greenhouse gases in the atmosphere, the occurrence of acid rains, climate
change, contamination of water basins, food contamination, loss of food security
requires development of a penalty system and urgent action, integrated and efficient
to solve global problems of the mankind, reducing and eliminating the negative
consequences of human impact on ecological systems. This requires the creation of
efficient mechanisms to minimize the negative impact of the economic activity on
ecological systems, the development of measures related to the implementation of
environmental management to restore ecological balance.
Pollution is characteristic to the modern, chemically intensive, mechanized
agriculture while air pollution is a common phenomenon in large cities and sur-
rounding areas. It has been found that in countries with a highly developed industry
air is polluted by transport (up to 50 %), by heating systems (18 %), by industrial
production (18 %). Considerable damages are caused by chemicals used in modern
agriculture, that don’t have analogy in nature and affect large areas. Environmental
disturbances caused by pollutants are manifested by decreasing the number and
diversity of plant and animal species by ecosystem reduction in size and degra-
dation, disruption of food chains, respectively, of the biocoenosis. On the other
hand, there is an uncontrolled multiplication of organisms (some insects, micro-
organisms) that adapt easily to these conditions due to the mechanisms of resistance
they possess.
Living organisms suffer directly or indirectly due to enhanced pollution of the
living environment. Thus the problem of environmental protection requires a
thorough study of the influence of different types of pollution on living organisms.
In this context it becomes particularly important to study the properties and
directions of the ongoing changes in natural systems, to unveil the adaptation
reactions and research the resistance mechanisms to unfavorable factors exhibited
by living organisms. Plants represent open systems that are in constant exchange of
matter and energy with the environment, they respond appropriately to adverse
conditions while the physiological and biochemical processes are perfectly coor-
dinated with the ambient factors. At the same time, pollutants generate in plants the
so-called stress conditions that represent the alteration of the processes of growth
10.7 Plant Resistance to Environmental Pollution 293
Fig. 10.13 Influence of heavy metals (Cd) on the cellular redox control (Schutzendubel and Polle
2002)
10.7 Plant Resistance to Environmental Pollution 295
Avoidance of heavy metal ion penetration into the plant cell is based on the
immobilization of heavy metal ions by the cell wall and removing the cell ligands that
act like chelating agents for metals. As a result, the plant avoids the toxic action of
heavy metals on intracellular processes. Immobilization and significant accumulation
of heavy metals happens in the free space of the cell wall or by binding metals to
specific sites of the extracellular matrix. Thus, in Agrostis tenuis, a plant tolerant to
Zn, there is a direct correlation between the ability of the cell wall to bind metal, and
the degree of resistance to the metal. Cultivation of Oryza sativa on media with toxic
concentrations of Cd and Zn and of Lupinus luteus on media with high concentration
of Cu and Pb led to the accumulation of these metals namely in the cell wall.
The accumulation of metal ions in the free space of the cell wall is determined by
the ion exchange coefficient, which depends largely on the number of histidine
groups in proteins and the number of carboxyl groups on the surface of pectic
substances from the cell wall. It is known that pectins play an important role in the
regulation of nutrient entry into the cell, including microelements. Carboxyl groups
create charges on the surface of pectins, which allows the retention of metal ions.
The higher the ion exchange coefficient, the more likely are toxic ions to be retained
in the gap of the cell wall, and vice versa.
The efficiency of heavy metal immobilization by the cell wall is also dependent
on the decrease of their activity after formation of stable chemical bonds with some
substances in the cell. It is assumed that heavy metals are bound by specific proteins
of the cell wall.
The plasma membrane serves as a selective barrier for heavy metal ions to enter
the cell. It can play an important role in the formation of plant resistance to heavy
metals, completely blocking the penetration of toxic ions into the cell.
The mechanisms involved in heavy metal detoxification and removal of these
ions from the cytoplasm is based on the property of organic compounds to bind
these metals. Plant tolerance to heavy metals may be provided by mechanisms that
participate in heavy metal detoxification and elimination from the cytoplasm, in
allowing plant cells to function normally in the presence of heavy metals or in
ensuring rapid restoration of damage caused by heavy metals.
Metal detoxification may be carried out by using organic acids, metallothioneins,
phytochelatins, glutathione and ferritins. Also to this type of mechanism is related
the active removal of heavy metal ions from the cell and appearance of tonoplast
transferases, capable to transport quickly metal ions from the cytoplasm into the
vacuole. All these mechanisms participate directly in binding or removing metal
ions from the cytoplasm.
Cosmic and terrestrial radiation on Earth existed long before the appearance of life.
The sum of all such cosmic and terrestrial irradiation form the global radiation
background.
296 10 Physiology of Plant Resistance to Unfavorable Environmental Factors
as compared to the aerial parts of the plant and the lateral roots are more sensitive
than the main, old leaves are more resistant than younger and so on, while the
pollen of the plants is more resistant than the oosphere, withstanding very high
doses (125–800 kR) of irradiation without losing germination capacity.
At the tissue level, meristematic cells and tissues, including those that divide
intensely are more sensitive than differentiated cells and tissues that divide slowly.
Specialized cells, such as stomata, are more resistant than the epidermis and the
latter, in turn, has a higher radioresistance than the meristematic tissue.
A different response to irradiation is recorded even in cellular organelles and
structures. The nucleus is more sensitive to radiation than the cytoplasm and
mitochondria. From the cell organelles, the strongest are the chloroplasts, which
undergo reduced changes even at lethal doses.
Radiosensitivity depends of the specificity of the irradiated biological material and
is determined by the seed size, the volume of the nucleus, the genetic nature of the
organism, size and number of chromosomes, DNA content and functional condition,
metabolic peculiarities of the species, the efficiency of the reparation systems etc.
The morphological and anatomical peculiarities mentioned above as well as the
physiological and biochemical particularities of the vegetal organisms are causing
the differences in the radiosensitivity among different species. Plant radiosensitivity
depends largely on the chemical composition of the cell, the metabolic particu-
larities of certain substances.
Among the biochemical components that provide increased plant resistance to
radiation are: fats, ascorbic acid, compounds with sulfide groups, growth sub-
stances, pigments “a”, oxidoreductases (peroxidase and catalase), etc.
Radioresistance is a genetically determined character, whose level of expression
depends on the internal conditions of the organism as well as on the environmental
conditions before and after irradiation (Fig. 10.14).
Fig. 10.14 Specific staining of lignin from the leaf of a mutant line of duckweed resistant to UV
radiation-mTR and sensitive to UV-PL 3FL7-11 (Jansen et al. 2001) phloroglucinol/HCI was
infiltrated in the branch with the leaves and it was studied under a light microscope. Mesophyll cell
walls are colored in red-orange.
298 10 Physiology of Plant Resistance to Unfavorable Environmental Factors
Enzymatic systems of restoration and DNA repair find the region damaged by
radiation, destroy it and restore the integrity of the DNA molecule.
At the cellular level there are some radioprotective substances: glutathione,
cysteine, ascorbic acid, metal ions and a number of enzymes and cofactors (cata-
lase, peroxidase, polyphenol oxidase, cytochromes C, NAD) which protect cells
from the damaging action of the radiation. The function of radioprotective sub-
stances is to neutralize free radicals arising from the action of the radiation, to create
local O2 insufficiency or to block reactions involving derivatives of radiochemical
processes.
At the level of the organism restoration is provided by: (a) the heterogeneity of
the population of meristematic cells; (b) asynchronous division in meristems, so
that at any time cells are involved in different mitotic stages characterized by
different radioresistance; (c) the presence in apical meristems of the mitotic back-
ground similar to the latent center, which starts to divide very actively when
division of the basic meristem stops which leads to the regeneration of both the
original cells and the meristem (d) presence of latent meristems of the type of
dormant buds which in the case of apical meristem destruction begin to function
actively and repair lesions.
Protection and repair mechanisms are not specific for plants only and hence, the
importance of studying them for solving the problem of radioresistance in both
plants and other living organisms.
Among the other types of resistance an important place has the resistance to high
concentrations of gases, which is the ability to maintain vital processes of plants
under the action of harmful gases (Polevoy 1982). The degree of plant resistance to
gases varies depending on the physical, geographical and meteorological
conditions.
It is known that the atmosphere composition varied over time along the evo-
lution of the organic world on earth. During abiogenesis Earth’s atmosphere con-
tained a number of compounds toxic for contemporary organisms (methane,
ammonia, hydrogen sulfide, carbon monoxide, etc.) which gradually changed with
the advent of life and due to other reasons as well. Nowadays, however, with the
rapid development of industry, atmospheric composition changes substantially in
record time. Presumably, the clearance of the primary earth atmosphere from these
gases was performed by early autotrophic plants, which are equipped with the
mechanisms of resistance to gas. Possibly, at that time, plants developed specific
mechanisms of resistance to toxic gases.
Contemporary flora formed in conditions when air pollution has occurred on
account of geological and chemical processes. As a result of the decrease of the
content of toxic gases, of the increase in the oxygen content of the atmosphere
(determining its oxidative character), organisms have partially lost their resistance
10.7 Plant Resistance to Environmental Pollution 299
and other stress factors, have a high resistance to gas, possibly due to their ability to
regulate the water regime and the ionic composition.
Resistance to gases can be increased by optimizing mineral nutrition and
hardening the plant material. The hardening is carried out by treating the seeds with
weak a solution of sulfuric acid or hydrochloric acid.
Air pollution affects differently the various species of trees and lower plants.
Lichens, trees like the elm, the ash tree and the pine are so sensitive to SO2 that may
serve as biological indicators of air pollution with this substance. Walnut and apple
trees are very sensitive to the harmful substances released into the atmosphere. In
these species drying of the tips of the branches and early flower detachment can
happen. In Quercus petraea and the black poplar Populus nigro significant defo-
liation and massive drying of the branches occur. Pelargonium zonale because of its
increased sensitivity to pollutants, can serve as an indicator of SO2 in the air by
presenting a yellowing of leaves, defoliation, pronounced chlorosis of the leaf blade
(at first on the edges, then progressing inwards along the leaf nerve). In Iris
germanica, Vitis vinifera, Lotus corniculatus burns on the leaves and leaf necrosis
caused by SO2 can be noticed which by reacting with water, produce H2SO3 and
H2SO4. Plant species such as tobacco are sensitive to oxidants and leaves of cab-
bage (Brassica)—to aromatic polycyclic hydrocarbons, as well as lead, iron,
chlorine.
Plants have an important role in cleaning the soil. Due to the absorption capacity
of the root system, a decrease in the content of all types of pollutants in the soil
happens. The plants are able to absorb from the soil nitroso compounds, oxides,
pesticides, heavy metals, etc.
Uptake of pollutants by plants occurs also in the aquatic environment. Aquatic
plants are involved in water purification and absorption of biogenic elements, of
hydrocarbons, phenols. In the Netherlands Scirpus is widely used for water puri-
fication. An active water purifier is Elchornia crassipes, which absorbs salts of
heavy metals, insecticides and detergents. Lemna absorbs heavy metals, nitrogen,
phosphorus, limiting their spread.
easily break down the side chain of the herbicide 2,4-D, which leads to its rapid
metabolization, whereas weeds are perishing as a result of accumulating the 2,4-D
herbicide, not due to its too high toxicity, but rather due to the excessive growth of
plants in a short time.
Crop species can detoxify from the 2,4-D herbicide either by esterification of its
carboxyl group with a monosaccharide either by forming a peptide bond with
glutamic acid. Detoxification is achieved also by hydroxylation of the aromatic
nucleus, followed by a reaction of glycosidation, or by oxidation of the side chain.
If the side chain of the herbicide is removed, it loses its hormonal activity and 2,4-
dichlorophenol is formed as an intermediate product, which by means of natural
glycosidation turns into the corresponding glycoside, which is not toxic for plants
(Fig. 10.15).
The xenobiotic transformation mechanism includes three main stages in the
detoxification of herbicides, insecticides and fungicides.
Phase I—at this stage reactions of degradation occur and of inclusion of new
functional groups in the initial structures, as a result of which the physical activity
changes as well as the hydrophilic or hydrophobic properties, the mobility of
substances, which reflects their ability to pass through the membrane and the
localization of the xenobiotics. Primary reactions are carried out with a very high
speed.
As a result of these reactions modified molecules have one or more groups with
high reactivity (−OH, −SH, −COOH, −sNH). Inclusion of new functional groups
allows further conjugation of the formed metabolites with other endogenous sub-
stances from plants.
Phase II (conjugation)—primary products are subjected to glycosidation,
esterification, conjugation with amino acids etc.
The conjugation reactions most frequently involve glucose. Reactions are pro-
duced by energy consumption in the presence of uridine diphosphoglucose (UDP-
glucose), under the action of glycosyltransferases. Depending on the nature of the
groups which reacted, O-glycosides, N-glycosides, and S-glycosides result.
Typically, glycosides and in particular the O-glycosides are highly labile substances
in vitro. In this context, their prolonged storage in plants would be impossible
without localizing them in cell compartments, where they are protected from the
action of hydrolytic enzymes. At the same time, by compartmentalizing the con-
jugates of the xenobiotics, their action on the metabolic processes of the cell is
10.9 Biochemical Mechanism of Pollutant Transformation in Plants 303
Fig. 10.16 Adaptation to abiotic stress (Arnholdt-Schmitt 2004). Typical phenotypic and
epigenetic changes at the level of the entire vegetal organism and at the cellular level in response to
phosphorus deficiency (a) and at the level of the factors involved in the global regulation of the
genome (b)
If the intensity of the factor does not exceed the lethal threshold, over a certain
time, the ratio of anabolic and catabolic products gradually returns to normal, and
initiation of the process of regeneration of the disturbed intercellular structures
306 10 Physiology of Plant Resistance to Unfavorable Environmental Factors
Glossary
References