BIOFERTILIZERS

INTRODUCTION
A large population of specific
beneficial micro-organisms which
enhance the fertility and
productivity of soil
They fix atmospheric Nitrogen or
solubilize soil Phosphorous
Based on renewable sources of
energy

 Very often microorganisms are not as

efficient in natural surroundings as one
would expect them to be and therefore
artificially multiplied cultures of
efficient selected microorganisms play
a vital role in accelerating the
microbial processes in soil.
Several microorganisms and their
association with crop plants are being
exploited in the production of
Biofertilizers. They can be grouped in
different ways based on their nature
and function.

TYPES OF BIOFERTILIZERS
1. Symbiotic Nitrogen Fixers: Rhizobium
2. Asymbiotic Nitrogen Fixers: Azatobacter
3. Algal Biofertilizers: BGA(BLUE GREEN ALGAE)
4. PSBS: Pseudomonas
5. Cellulolytic Bacteria/Fungi
6. Mycorhiza : VAM

NITROGEN FIXING
BIOFERTILIZERS
Nitrogen is one of the main components
of living cells. Even though the
atmosphere constitutes of Nitrogen as
the major gas living organisms are
unable to use Nitrogen as it exists in the
air.
Nitrogen combines with Hydrogen to
form Ammonia and with Oxygen to
form Nitrates

Biological Nitrogen Cycle

Symbiotic Nitrogen Fixers

Symbiotic nitrogen fixation occurs in plants
that harbor nitrogen-fixing bacteria within
their tissues. The best-studied example is
the association
between legumes and bacteria in the
genus Rhizobium.
Each of these is able to survive
independently (soil nitrates must then be
available to the legume), but life together is
clearly beneficial to both. Only together can
nitrogen fixation take place.

The bacteria enter an epithelial cell of the root; then migrate into the
cortex
. The cortex cells then begin to divide rapidly forming a nodule.
The Rhizobium also go through a period of rapid multiplication
within the nodule cells. Then they begin to change shape and lose their
motility. The bacteroids, as they are now called, may almost fill the
cell. Only now does nitrogen fixation begin.
Root nodules are not simply structureless masses of cells. Each
becomes connected by the xylem and phloem to the vascular system of
the plant.
The legume is certainly helpful in that it supplies nutrients to the
bacteroids with which they synthesize the large
amounts of ATP needed to convert nitrogen (N2) into ammonia (NH3)
In addition, the legume host supplies one critical component
of nitrogenase the key enzyme for fixing nitrogen.

Asymbiotic Nitrogen Fixers

Asymbiotic nitrogen fixation is of great importance
agriculturally.
Some bacteria which are important non-symbiotic
bacteria are Acetobacter, Achromobacter,

Azospirillum, Clostridium, Enterobacter,

Pseudomonas, Rhodospirillum, Xanthobacter etc.

Diazotrophic bacteria like species of Azospirillum,

Azotobacter, Clostridium and Bacillus are commonly
associated with higher plants. The dinitrogen fixing
bacteria are widespread and it has led to interest in
determination of their importance in agriculture. The
contribution of asymbiotic nitrogen is a dominant
source in terrestrial ecosystems.

Nitrogen Fixation by

Rhizobium

Infection thread with Nodule formation

Azotobacter
The Azotobacter is of the family Azotobacteraceae, it is a cyst forming
species and is generally regarded as a free-living aerobic nitrogen fixing
bacteria. The Azotobacter s are large Gram negative bacteria, they are
obligatory aerobic rods, and they are capable of fixing nitrogen
asymbiotically. The application of Azotobacter and Azospirillum improves
the yields of annual and perennial grasses.

Azospirillum
The members of this genus are mostly associated with the roots and
rhizospheres of large number of agriculturally important crops and cereals.
Azospirillum can enhance the growth of plants and yield a wide range of
economically important crops in different types of soil and climatic
conditions. The beneficial aspects of Azospirillum are for the production of
phytohormones, reduction of nitrate and nitrogen fixation. They are known
to be associated with diazotrophs. Despite the property of nitrogen fixation
they also attribute in the development of root as they produce growth
promoting substances and increase rate of water and mineral intake.

Azotobacter colonies

ALGAL BIOFERTILIZERS
These help in Nitrogen fixation.
BGA are photoautotrophic,
prokaryotic algae which are highly
recommended as Biofertilizers.
These include both unicellular as well
as filamentous forms. Nostoc,
Anabaena are some of the filamentous
forms which help in N2 fixation.

Many cyanobacteria fix atmospheric nitrogen, and a large

number of them do so under aerobic conditions. The nitrogen
sources most commonly used by cyanobacteria are nitrate,
ammonium, urea, and dinitrogen.
Cyanobacteria confine nitrogenase to heterocysts,
differentiated cells specialized in nitrogen fixation, some
other, unicellular as well as filamentous, strains express the
nitrogenase activity in the dark periods of light-dark growth
cycles.
Heterocyst-forming species are specialized for nitrogen
fixation and are able to fix nitrogen gas
into ammonia, nitrites or nitrates , which can be absorbed by
plants and converted to protein and nucleic acids
Cyanobacteria fulfill vital ecological functions in the world's
oceans, being important contributors to global carbon and
nitrogen budgets.

BGA Culture

PHOSPHATE SOLUBILIZING
BACTERIA AS BIOFERTILIZERS

The use of phosphate solubilizing bacteria as inoculants

simultaneously increases P uptake by the plant and crop yield.

Strains from the genera Pseudomonas, Bacillus and Rhizobium are

among the most powerful phosphate solubilizers.

The principal mechanism for mineral phosphate solubilization is

the production of organic acids, and acid phosphatases play a
major role in the mineralization of organic phosphorous in soil.

Genetic manipulation of phosphate-solubilizing bacteria to

improve their ability to improve plant growth may include
cloning genes involved in both mineral and organic phosphate
solubilization, followed by their expression in selected
rhizobacterial strains

Phosphate solubilization takes place through various microbial

processes / mechanisms including organic acid production and proton
extrusion.
Phosphorus solubilization is carried out by a large number of
saprophytic bacteria and fungi acting on sparingly soluble soil
phosphates, mainly by chelation-mediated mechanisms.
The PSB dissolve the soil P through production of low molecular
weight organic acids mainly gluconic and keto gluconic acids, in
addition to lowering the pH of rhizospheres.
In certain cases phosphate solubilization is induced by phosphate
starvation

USES OF PSB
Encourages early root development
Fosfofix produce organic acids like malic, succinic,
fumaric, citric, tartaric and alpha ketoglutaric acid
which hastens the maturity and thereby increases the
ratio of straw as well as the total yield.
Increases compatibility of the other beneficial
microbes with the plants
Stimulates formation of fats and convertible starches
Helps in rapid cell development in the plants and
consequently increases the resistance towards disease

Culture of Phosphobacteria

CELLULOLYTIC BACTERIA
AND FUNGI
Cellulolytic Bacteria and Fungi are the most
important microorganisms for the
decomposition of organic matter due to
their degradation abilities. When large
amounts of extracellular enzymes, essential
for the degradation of substrates, such as
lignocelluloses, are produced, the
decomposition of this organic matter can
occur.

By doing this they enrich the nutrient

capacity of the soil thus acting as excellent
Biofertilizers.

Sporangia of anaerobic
fungi(electron microscope)

MYCORHIZA AS
BIOFERTOLIZERS

Mycorrhiza is the product of an association between a fungus and

plant root. Vesicular-arbuscular mycorrhiza (VAM) is formed by
the symbiotic association between certain phycomycetous fungi
and angiosperm roots

The fungus colonizes the root cortex forming a mycelial network

and characteristic vesicles (bladder-like structures) and
arbuscules (branched finger-like hyphae).

The arbuscules are the most characteristic structures, formed

intracellular and probably having an absorptive function. The
vesicles are terminal swellings of hyphae formed inter and
intracellularly having a storage function

Glomus, Gigaspora, Acaulospora, Entrophospora Sclerocystis and

Scutellospora are some examples.

VAM as Biofertilizers
Mycorrhiza plays a very important role
on enhancing the plant growth and
yield due to an increase supply of
phosphorus to the host plant.
Mycorrhizal plants can absorb and
accumulate several times more
phosphate from the soil or solution
than nonmycorrhizal plants. Plants
inoculated with endomycorrhiza have
been shown to be more resistant to
some root diseases

Schizosaccharomyces octosporus

Commercial production of
biofertilizers
The contamination of the soil due to excessive use of
chemical fertilizers and pesticides is one of the major
problems facing agriculture. Hence the alternative is use
biological nitrogen fixation technology for maintenance of
soil health and sustainable agriculture

At present Biofertilizers using microorganisms are

produced in large scale industrially and are available for
field application.

Biofertilizer production technology comprises three

important steps:
1) development of strains
2) upscale of biomass
3) preparation of inoculants

CULTURE MEDIA
Rhizobium : Yeast extract mannitol broth. Mannitol10.0 gK2; HPO4 -0.5 g; MgSO4 7H2O-0.2 g; NaCl-0.1 g;
Yeast extract-0.5 g; Agar 20.0 g; Distilled
water 1000.0 ml
Azotobacter :Waksman medium No.77 (N-free
Mannitol Agar Medium for ) Mannitol:10.0 g;
CaCO3 :5.0 g; K2HPO4:0.5g; MgSO4.7H2O:0.2g;
NaCl:0.2 g ; Ferric chloride: Trace
MnSO4.4H2O:Trace N-free washed Agar:15.0 g ;
pH:7.0 Distilled Water:1000 ml

STEPS
Pure bacterial broth with high cell count is blended
aseptically with sterilized carrier such as peat,
charcoal and/or lignite so as to obtain a moist
powdered formulation having high population of
desired microbes
Bacterial strains are usually developed and
maintained by research laboratories

For mass production of Rhizobium and Azotobacter in

pilot scale biofertilizer technology the following
equipments are required: (1) Twenty liter fabricated
fermenter fitted with an air pressure pump, (2)
horizontal rotary shaker, (3) vertical autoclave with
pressure regulator, (4) incubator, (5) refrigerator, (6)
inoculation chamber (with provision of UV-C), (7)
distillation apparatus, (8) microscope, (9) polythene
sealer and (10) air-conditioning provision

Region specific and environmental stress compatible

bacterial strains isolated from various agroclimatic regions
are obtained from authentic sources, grown in slants and
transferred to liquid broth in the rotary shaker to prepare
mother culture. The cultures are grown in large scale in
the fermenter for up to five days, harvested in batch
culture mode and then mixed with unsterile forest soil:
charcoal in a ratio of 1:3.
The shade dried bacteria and the carrier
are mixed and packed, 200 g each in polythene pack, and
stored for use as biofertilizer for the desired crop
. Quality of inoculants in the biofertilizer pack is one of the
most important factors resulting in their success or failure
and acceptance or rejection by the farmers. Basically the
quality means the presence of right type of
microorganism in active form and in desired numbers.

Production of Biofertilizers

MODE OF Application of
Biofertilizers
Seed treatment
One packet of the inoculants is mixed with 200 ml of rice kanji to
make a slurry. The seeds required for an acre are mixed in the
slurry so as to have a uniform coating of the inoculants over the
seeds and then shade dried for 30 minutes. The shade dried seeds
should be sown within 24 hours. One packet of the inoculants
(200 g) is sufficient to treat 10 kg of seeds.

Seedling root dip

This method is used for transplanted crops. Two packets of the
inoculants is mixed in 40 liters of water. The root portion of the
seedlings required for an acre is dipped in the mixture for 5 to 10
minutes and then transplanted.
Main field application
Four packets of the inoculants is mixed with 20 kgs of dried and
powdered farm yard manure and then broadcasted in one acre of
main field just before transplanting.

LIQUID BIOFERTILIZERS
1.Contains special cell protectants or substances that encourage
formation of resting spores or cysts.
2. Contains special nutrients that ensure longer shelf life, Better
survival on seeds and soil and tolerance to adverse conditions.
3. Liquid formation ensures that the product is easy to handle and
apply.

4. Since the organisms are stabilized during production, distribution

and storage, the activity is enhanced after the contact and interaction
with the target crops.
5. No loss of properties due to storage up to 55 degrees.
6. Greater potentials to fight with native population.
7. Dosages are 10 times lesser than the carrier-based bio-fertilizers.
8. Very high enzymatic activity since contamination is nil.

USES OF BIOFERTILIZERS IN
AGRICULTURE

They supplement chemical fertilizers for meeting the integrated

nutrient demand of the crops.

They can add 20-200 kg N/ha year (e.g.. Rhizobium sp 50-100 kg

N/ha year ;Azospirillum , Azotobacter : 20-40 kg N/ha /yr; Azolla :
40-80 kg N/ha; BGA :20-30 kg N/ha) under optimum soil
conditions and thereby increases 15-25 percent of total crop yield.

They can at best minimize the use of chemical fertilizers not

exceeding 40-50 kg N/ha under ideal agronomic and pest-free
conditions.

Application of Biofertilizers results in increased mineral and

water uptake, root development, vegetative growth and nitrogen
fixation.

Some Biofertilizers (e.g., Rhizobium BGA, Azotobacter sp)

stimulate production of growth promoting substance like
vitamin-B complex, Indole acetic acid (IAA) and Gibberellic acids
etc.

USES

Phosphate mobilizing or phosphorus solubilizing Biofertilizers /

microorganisms (bacteria, fungi, mycorrhiza etc.) converts
insoluble soil phosphate into soluble forms by secreting several
organic acids and under optimum conditions they can solubilize /
mobilize about 30-50 kg P2O5/ha due to which crop yield may
increase by 10 to 20%.

Mycorrhiza or VA-mycorrhiza (VAM fungi) when used as

Biofertilizers enhance uptake of P, Zn, S and water, leading to
uniform crop growth and increased yield and also enhance
resistance to root diseases and improve hardiness of transplant
stock.

They liberate growth promoting substances and vitamins and

help to maintain soil fertility.

They act as antagonists and suppress the incidence of soil borne

plant pathogens and thus, help in the bio-control of diseases.

Nitrogen fixing, phosphate mobilizing and cellulolytic

microorganisms in bio-fertilizer enhance the availability of plant
nutrients in the soil and thus, sustain the agricultural production
and farming system.

Azolla
Azolla is a free floating water fern that
floats in water and fixes nitrogen in
association with the nitrogen fixing blue
green algae, Anabaena azollae. Azolla is
considered to be a potential biofertilizer in
terms of nitrogen contribution to rice. Long
before its cultivation as a green manure,
Azolla has been used as a fodder for
domesticated animals such as pigs and
ducks. In recent days, Azolla is very much
used as a sustainable feed substitute for
livestock especially dairy cattle, poultry,
piggery and fish.

Advantages of biofertilizers
High yield of crops are obtained.
They are eco-friendly.
They produce better yields than
chemical Biofertilizers
They contain natural products which
are harmless to the plant
They are cost effective
They are easy to handle

Precautions to be taken
The bacterial inoculants should not
be mixed with insecticides,
fungicides, herbicides, etc.
The inoculum treatment should
always be the last or the final step

Constraints or
disadvantages
Though the biofertilizer technology is a
low cost, eco-friendly technology,
several constraints limit the
application or implementation of the
technology the constraints may be
environmental, technological,
infrastructural, financial, human
resources, unawareness, quality,
marketing, etc. The different
constraints in one way or other
affecting the technique at production,
or marketing or usage.

Technological constraints
Use of improper, less efficient strains for
production.

Lack of qualified technical personnel in

production units.
Unavailability of good quality carrier material or
use of different carrier materials by different
producers without knowing the quality of the
materials.
Production of poor quality inoculants without
understanding the basic microbiological
techniques
Short shelf life of inoculants.

Infrastructural constraints
Non-availability of suitable facilities for
production
Lack of essential equipments, power supply,
etc.
Space availability for laboratory,
production, storage, etc.

Lack of facility for cold storage of inoculant

packets

Financial constraints and

Environmental constraints
Non-availability of sufficient funds and problems in
getting bank loans
Less return by sale of products in smaller production
units.
Seasonal demand for biofertilizers
Simultaneous cropping operations and short span of
sowing/planting in a particular locality
Soil characteristics like salinity, acidity, drought,
water logging, etc.

Awareness on the technology

and Marketing constraints
Unawareness on the benefits of the technology.
Problem in the adoption of the technology by the farmers
due to different methods of inoculation.
No visual difference in the crop growth immediately as that
of inorganic fertilizers.
Unawareness on the damages caused on the ecosystem by
continuous application of inorganic fertilizer.
Non availability of right inoculant at the right place in
right time.
Lack of retain outlets or the market network for the
producers.

conclusions
The use of biofertilizers is a
promising future for a healthier
environment and thus their use
should be made practical and
frequent. They are healthy, ecofriendly and thus are pollution free
and harmless.

Presentation by:
Deesha r

Tenzing n

Sreeja Nair

ESTHER JAMIR

Sharvarika poovaiah

SUSHMITHA N

Harshita r

MARTINA

Shruthi n

RABIA A

Kibria mariam

SYEDA AFREEN

Twinkle s

AFREEN BANO

ARCHITHA S

THANK YOU