Plant Physiology I:

Introduction:
Cell division, expansion, cell-cell communication:
Cell division:
How does plant organ-shape arise?
Organization of cells creates strict geometric shapes.
Formative vs proliforative cell division in the plant root.
- Formative cell division serves to increase the number of cell files in the root.
- Proliferative division serves to extend the length of an existing file of cells in the
root. (Fig. 1).

Formative
cell division

Proliforative
cell division

Figure 1: Two different directions of cell division.

-

1) Organismal theory of plant development Expansion leads, mitosis follows:

- Cell size homeostasis: Anisotropic expansion of plant organs results in surfacevolume ratios in excess of efficient transport and cell metabolism. The overly
large cell divides in order to partition the existing cell volume between two,
smaller cells, both being run more efficiently than the one large cell.
-

Support for Expansion leads, mitosis follows:

-

Within a given plant tissue, cellular volume and surface area serviceable by a
single nucleus are limited and strictly correlated with the C-value (number of
haploid genomes) in the nucleus.

In yeast, mutations permitting the early entry into the cell cycle produces
abnormally small daughter cells. These cells then delay the next round of
mitoses until they have grown to approximate wild-type cell size. In plants,
the larger of two daughter cells resulting from unequal cytoplasmic
partitioning re-enters the cell cycle before the smaller cell.

Ethylene stimulated elongation of deepwater rice plants results from GA

stimulation of pre-existing cells to elongate. Later, after the cells have
elongated considerably, they commence division.

2) Mitosis leads, expansion follows:

-

Support for Mitosis leads, expansion follows:

-

Experiments have shown that in transgenic plants ectopically expressing

Mitotic cyclin cells are driven into the cell cycle, forcing them to divide. These
artificially replicated cells then expand. The Mitosis leads, expansion
follows scenario seems to suggest that the default for a newly divided cell
is.expansion.

The same yeast mutants mentioned above provide an argument FOR mitosis
leads, expansion follows! Due to the mutation, the yeast cells divide
producing smaller cells which then "catch up" to the wild type by expansion.

Additionally, GA has been suggested to be a mitogen, initiating mitosis.

How do you stop dividing?:

In vertebrate cells, withdrawal from the cell cycle is mediated by small
proteins that inhibit cyclin-dependent kinases (CDKs), proteins involved in
priming the cell for DNA synthesis, leading to mitosis. Hence, the cells become
"trapped" in the G1 resting phase. This fits well with the knowledge that the
mutations in yeast mentioned above that permit premature entry into the cell
cycle resulting in smaller than usual daughter cells negate a tyrosine-kinase
responsible for phosphorylating a tyrosine on CDK1, leading to its inactivation.
Hence, in the yeast mutant, CDK1 is not inactivated and this leads to premature
mitosis and smaller cells.
In plants cells there is evidence to suggest that the same mechanism
may be in place. Cyclin-dependent kinase inhibitors (CDIs) have been
identified in maize endosperm arrest.

Synthesis: There may be two mechanisms at work, morphogenetic (formative) cell

divisions are overriding more fundamental "default" divisions (proliferative) that
are the default for expanding cells to maintain cell size homeostasis. Thus, the
large variation in plant organ shape arise from selectively stopping cell division
(formative) in some cells.
Cell expansion:
Water uptake drives cell expansion in plants which is an irreversible increase in cell volume. By
taking up water into the central vacuole, plants have developed an economical method of
increasing their size by orders of magnitude while maintaining approximately the same
cytoplasmic volume, albeit, now dispersed over the periphery of a larger cell. Water uptake into
the vacuole to increase turgor, will not be effective in driving cell expansion unless the rigid cell
wall is somehow induced to weaken, allowing the pressure within to force the cell wall to extend.
This is not analogous to air forcing a balloon to expand since the wall of the balloon becomes
increasingly thin until it ultimately fails. The plant cell wall deposits and incorporates new wall
material into the expanding cell wall so that no wall thinning occurs. The ability of the cell wall to
expand and incorporate new cell wall material while under stress is the result of its complex
makeup and precise partial disassembly by some select wall modifying enzymes.
There are five main components to plant cell walls which in dicots have the following approximate
percent compositions; 1) cellulose (~30% wall dry weight); 2) hemicelluloses (~30% wall dry
weight); 3) pectins (~35% wall dry weight); 4) proteins (~1-5% wall dry weight) and; 5) water
(~75% wall wet weight). Ions such as calcium make up a sixth component but in truly negligible
amounts relative to the first 5.
Cellulose is the backbone of the cell wall, imparting to it its ability to withstand expansion, or to
expand and dictate the directionality of this expansion. Cellulose in the cell wall is a rigid semicrystalline polymer that is laid down on the inner cell wall surface by a complex assemblage of
proteins arranged in a rosette often referred to as the terminal rosette since it is found at both
termini of elongating microfibrils. The rosette is embedded in the plasmamembrane spanning it
and

presumably anchored to and travelling along actin polymers aligned on the inner surface of the
cell membrane. How the cellulose microfibrils are arranged determines if and how the cell
expands. If the microfibrils are secreted in a non-parallel manner (isotropically), then the
microfibrils may prevent cell expansion since there is no direction the cell may extend without
breaking a microfibril (Fig.2). To date, there have been no reports of cellulases produced by

This figure is of a plant cell wall in

w h i c h t h e c e l l u l o s e m icrofibrils h a v e
b e e n l a i d d o w n i s o t r o p i c a l l y. T h e
m icrofibrils w ill either resist cell
expansion in any direction or slip past
each other allowing equal cell
enlargement in all directions at once in
a p r o c e s s c a l l e d p o l y m e r c r e e p .

Isotropic or
no growth

In this figure, during plant cell

enlargement, control of the direction of
cell growth is imposed by arranging
t h e m icrofibrils p e r p e n d i c u l a r t o t h e
d i r e c t i o n o f g r o w t h . T h e m icrofibrils
resist expansion laterally much more
than they do longitudinally (in the
direction of the arrows). This control of
plant cell expansion is crucial to plant
development since it is the direction
and amount of cell expansion that
determines final organ shape and size
in plants.

Anisotropic
(directional)
growth
Cellulose
Hemicellulose
Pectin
Cell wall proteins
Figure 2: Diagram of the plant cell wall from two different cells, one that is not expanding or
expanding isotropically and a second expanding directionally or anisotropically.
plants that are capable of hydrolyzing crystalline microfibrils. The cellulases that have been
discovered are all thought to act on hemicellulose substrates such as xyloglucan. Alternatively, in
response to internal signals, actin filaments can arrange themselves in parallel with the result that
the enzyme rosettes extruding cellulose microfibrils to the inner wall surface exterior to the
plasmamembrane lay down the microfibrils in parallel (anisotropically, Fig. 2). Hence, the
configuration of the cellulose in the stress bearing region of the cell wall resembles a spring or
slinky. It is difficult to expand the spring laterally but much easier to extend the spring
longitudnally.
The hemicellulosic component of plant cell walls is thought to be intimately associated with the
cellulose microfibrils, either laying along the microfibrils for much of their length associated by

hydrogen bonds, or actually embedded in the interior of the microfibrils. The hemicellulosic
constituents are delivered to the cell wall from Golgi derived vesicles in which they are
synthesised. Upon arrival at the plasma membrane the vesicles fuse with the membrane and
release the contents into the wall. These wall components are then somehow incorporated into
the stress bearing wall, probably being linked together enzymatically to form longer, more highly
branched polymers than were synthesized in the Golgi. This process appears to be operational in
the deposition of xyloglucan into the cell wall of plants using xyloglucan endo-transglycosylase
(XET) to polymerize short, vesicle-deposited xyloglucan chains into larger polymers of
xyloglucan. Additionally, the simultaneous extrusion of semi-crystalline cellulose at the inner wall
surface and a marked affinity of xyloglucan for cellulose ensures their tight association, even the
possibility of xyloglucan embedding itself into the elongating microfibril. To permit cell wall
expansion, these hemicelluloses must be either disassociated from the cellulose microfibrils and
themselves by a recently discovered enzyme christened expansin or disentangled from the
matrix usually though partial polymer hydrolysis by endo-glycosylases (EGases).
Pectin forms a gel in which the other cell wall components are embedded. Although it is usually
thought of as existing in the middle lamella between cells, cementing them together, pectin is
found throughout the plant cell wall. Pectin is initially deposited at the inner cell wall surface in
discrete lengths by Golgi derived vesicles where it is formed. This pectin is highly methyl
esterified when deposited making it resistant to hydrolysis and relatively unreactive with cell wall
calcium and other ions. Upon deposition, the pectin is typically de-esterified by pectin methyl
esterase (PME) imparting to it a negative charge and allowing it to form rigid configurations and
bind tenaciously to calcium. It is also now susceptible to modification by pectin hydrolysing
enzymes (polygalacturonases, PGs). The degree to which pectin is hydrated, its degree of
esterification, and the bonds it is participating in are all thought to control its porosity which in
turn determines the sieve size of the cell wall, dictating what size particle, protein or
polysaccharide, can pass through the matrix.
Protein in the cell wall can be structural, such as extensins thought to impart inflexibility to the
cell wall, or enzymatic, such as XET, EGases, PME, PG, etc. Although they comprise only a
small percentage of the total cell wall by dry weight, they are absolutely crucial to its ability to
expand.
Water is often ignored as a major component of the living, dynamic plant cell wall but, besides
imparting solvent and lubricational properties, it also determines the concentration of the
components of the wall and their hydration determines wall porosity. Its influx into the vacuole of
cells dictates the turgor they are capable of exerting and no plant cell, however disposed to cell
wall loosening, can expand without turgor pressure.
Plant cell walls isolated from regions undergoing natural expansion are susceptible to elongation
due to a decrease in pH, so-called acid growth. Protease treatment has been shown to eliminate
acid growth from plant cell walls normally responding to decreased pH. This led to Daniel
Cosgrove isolating two proteins from cucumber hypocotyls that potentiate acid growth. These
were named expansins and are purportedly soley responsible for acid growth of plant cell walls
without cleaving any cell wall constituent. The expansin-mediated growth of plant cell walls is
enhanced by pretreatment of cell walls by hemicellulases and presumably this also is the case in
vivo, but the hemicellulases tested to date contribute little to cell wall expansion relative to
expansin. Yet, in plant parts that normally do not expand, the cells are resistant to acid-mediated
elongation. There is therefore, a point in the life of a cell where it becomes rigidly fixed in size and
unresponsive to molecules and environments that promoted cell elongation in the tissue at some
earlier developmental stage. This is termed rigidifying the cell wall and generally fixes cell size
irreversibly. The process of regidification is thought to involve the reduction of cell wall loosening
processes, a more comprehensive cross-linking of cell wall components resisting cell wall
expansion, and an alteration in the components of the cell wall, stiffening it against extension.
Cell-cell communication:

Cell-cell communication in plants includes the regulation of plant growth and development by
phytohormones, the receptor-ligand signaling evident in pollen-stigma interactions leading to selfincompatibility responses, and intercellular trafficking of substances through plasmadesmata
(PD). PD are unique to the plant, a consequence of how cell division occurs in these organisms.
The fact that; 1) there is incomplete separation of the cytoplasm during cytokinesis and; 2) a cell
plate forms de novo across the cell except for regions of cytoplasmic continuity between the two
daughter cells, permits the cytoplasmic continuity between the cell clones. Hence, every set of
daughter cells are symplastically continuous through what has become known as primary,
unbranched PD. Cells that are not derived from the same mother cell can also form PD
connections and do so frequently. The mechanism by which these PD are formed varies from that
of primary PD in that secondary branched PD can form de novo in preexisting cell walls. They
usually consist of multiple cytoplasmic strands that converge between the two cells in a central
cavity before branching into multiple channels again to enter the second cell.
Symplastic domains: The cells of the embryos of those plants studied to date have been shown to
be all symplastically continuous. However, soon after the completion of germination, this
symplastic continuity is disrupted so that assemblages of cells in the seedling lose their PD
connections with other assemblages. This establishes groups of cells that, although in direct
symplastic contact with each other, are isolated symplastically from additional groups of cells
setting up domains of cells that, although in close proximity to each other may be in very different
physiological states. The symplastic continuity of cells and their isolation from neighboring cells
has vast implications for plant cell differentiation which is dependent on cell position. Evidence
from cell ablation studies has shown that more mature cells in the plant root tip dictate to recently
divided cells more distal in the same file, their developmental fate. Moreover, there exists a
mechanism to dictate the direction of propagation of such a signal so that laterally adjacent cells
are unaffected by signals produced in the more mature cell, only those cells within the same file
and less mature than the cell producing the signal are influenced. One mechanism allowing this
type of developmental control would be to have the cells within a file, all clones of their initial at
the root apex, symplastically continuous with each other through primary PD and symplastically
isolated from adjacent files of cells. This is only a hypothesis but is a possible explanation for this
developmental phenomenon. In support of this hypothesis are two observations made with
diffusable dyes. The first is that the more mature cells of the arabidopsis epidermis are
symplastically isolated from the underlying cells of the cortex. Additionally, root hairs are also
symplastically isolated from the epidermal cells surrounding them. A mutant in cell wall
development (knolle) artificially maintains symplastic connections between symplastic domains
not normally associated. As a probable result, the knolle mutant has many abnormalities in
development. The guard cells comprising the stomata are completely symplastically isolated by
the time the stomata are completely differentiated. The PD connections between the guard cells
and their surrounding cells are lost though protein degradation of PD components.
Plasmodesmatal connections can also undergo developmental changes during the differentiation
of plant cells. In young buds and flowers of Setcreasea purpurea the PD size exclusion limits
(SEL) of stamen hairs are less than 1000 Daltons while in senescing flowers, the SEL is greater
than 4.4 kD. SEL also increases due to environmental stimuli. Anoxia increases SEL in wheat
roots and osmotic stress increases SEL in pea roots while changes in light quality alters
symplastic trafficking rates in maize. Additionally, turgor differences between adjacent cells can
close existing plasmadesmata so that the cells are symplastically isolated despite the existence
of plasmadesmata. Differences in turgor pressure gating plasmadesmata is a sound strategy to
prevent catastrophic dehydration should one cell in a symplastic domain become injured.
Plasmadesmata are also closed by callose plugs in the case of injury to the cell.
The developmental changes that regulate PD function and SELs has profound implications for
plant-virus interactions. Plant viruses have been shown to be capable of altering the SEL of PD
considerably. However, this ability to alter SEL is modified by host developmental stage and
physiological state. Hence, arabidopsis plants grown under long-days mature early and are

resistant to the cauliflower mosaic virus (CaMV) while those grown under short-days mature
slowly and are not as resistant to CaMV infection. By anology, if pathogenic molecules/organisms
can move systemically though PD, then it is anticipated that endogenous protective molecules, so
called pathogen related (PR) molecules, may also turn out to be targeted to PD for further
transport into neighboring cells.
Plasmadesmata are complex organelles, certainly not a simple "hole-in-the-wall".

Closed
Plasmodesmata

Extracellular
sphincter

Open
Plasmodesmata

Neck
Central
cavity

Cell wall

Plasmamembrane
Desmotubule

Cytoplasmic
sleeve

Redrawn from The Plant Cell 9: 1047.

Figure 3: Diagramatic representation of one model of plasmodesmata structure.
The cytoplasmic sleeve of plasmodesmata appears to be subdivided into many smaller
microchannels (Fig. 3). The "spokes" detected in the lumen of the plasmodesmata are probably
comprised of actin, a protein that has been localized to the PD. Alterations in the size exclusion
limit of PD has led to intensive study of various viral and plant proteins thought to function in the
modification of the SEL of the PD. These proteins are capable of transporting select, very large
proteins and nucleic acid polymers through the plasmodesmata. These so called movement
proteins are currently under intense scrutiny to determine how, mechanistically, they permit such
trafficking. With their discovery has come the realization that the PD probably play a hither to
vastly undervalued role in cell-to-cell communication in plants.
Models of transport through PD: Cell biologists use inert, usually fluorescent, dextrans of
precisely defined size to determine the SEL of plasmodesmata. Using these dextrans in concert
with plant or viral movement proteins has revealed some interesting results regarding the
probably method of transport through PD. Evidence exists that there is more than one type of
protein facilitated movement through PD. Relatively short distances between mesophyll cells
leads to short PD which, upon being dilated by movement proteins in the presence of large
dextran molecules, permit the transport of both since the whole route is gated open

simultaneously. However, longer PD linking trichome cells with other cells may either; 1) be
capable of unfolding proteins with associated transport signals and transporting them through the
undilated PD to refold on the other side or; 2) gate the PD open very quickly and propagate the
open state of the PD along the PD so that the PD is not open from neck to neck simultaneously in
response to a movement protein signal. Either scenario results in the trafficking of the movement
protein only and the exclusion of the dextrans from transport into neighboring cells unlike
movement through PD in mesophyll cells.