Biofertilizers

Definition
Biofertilizers includes selective microorganisms
like bacteria, fungi, algae which are capable of
fixing atmospheric N or convert insoluble
phosphate and other salts in the soil into forms
available to plants.
Photosynthesis: CO2 + H2O  carbohydrates (CHO) + O2

Major Cause: Nutrient

Deficiency In Soil

Nutrients are taken up primarily by the

roots in the form of an aqueous solution
in the soil
Why Biofertilizers???
Disadvantages of chemical fertilizers:
• The chemical fertilizers are used for better crop yield which
are also providing N,P.
• But use of these fertilizers in excess of recommended levels in
order to ensure high yields(generally farmers do),cause
environmental pollution.
• These environmental contamination & over supply of
nutrients can lead to negative consequences on humans and
animals.
- Ingestion of nitrate can be toxic to humans.
• It cause eutrophication.
Advantages
• Biofertilizers are the microbial inoculants
contain unique and beneficial strains of soil
microbes.
• The Biofertilizers production cost is very low
• On nutrient basis 1 tonne of Rhizobium
Biofertilizers is equivalent to 100 tonne of
fertilizer nitrogen (Verma & Bhattacharyya,
1991).
Advantages
1. Biofertilizers are supplements to chemical
fertilizers.
2. They are cheap & can help to reduce fertilizer
consumption.
3. They provide biological nitrogen directly to plants
4. They help in solubilization & mineralization of
other plants nutrients like phosphates.
5. They enhance plant growth due to release of
hormones, vitamins, auxins and other growth
promoting substances.
Advantages
6. On an average crop yield increases by 10-20
percent with their use.
7. They control and suppress soil borne diseases
8. They help in the proliferation and survival of
beneficial micro organisms in the soil
9. They improve soil properties and maintain soil
fertility.
10. They are eco-friendly and pollution free.
Types of biofertilizers
• Nitrogen fixers
1. Symbiotic: Rhizobium, Frankia
2. Non symbiotic: Azotobacter, Azospirillum, Blue
green algae, Azolla, Acetobacter

• Phosphate supplier
1. Phosphate solubiliser: Bacillus, Pseudomonas,
Aspergillus
2. Phosphate absorber: V.A.Mycorrhiza (VAM fungi)
Types of biofertilizers
• Sulphur supplier
1. Thiobacillus novellus, Aspergillus

• Organic matter decomposer and microbial cell

mass.
1. Cellulose decomposer, Lignin decomposer.
Comparative overview of chemical fertilizer and Biofertilizers
Factors Chemical fertilizers
production Industrial, centralized
Process chemical
Raw materials Fossil fuel & others imported, based on
and energy budget non-renewable energy sources, energy
intensive
Efficiency 40-45% for nitrogenous fertilizers for
upland crops and less than 33 % for
rice,25-33% or phosphatic fertilizers.
Heavy N losses due to leaching,
volatization and denitrification.P
availability decreased due to fixation.
Effect on For nitrogen- nil or low
subsequent crop
Pollution effect Exists due to discriminate use
Biological fertilizers
Biological, small scale or
decentralized
Biological
Atmospheric nitrogen for nitrogen
fixers, unavailable P for PSM and
VAM, Organic residues for compost
decomposing organisms.Indigeous
and local, based on renewable
energy sources very low energy bill.
About 90% efficiency. Losses due to
leaching, fixation are negligible.
Residual effect for nitrogen
Pollution free
 Rhizobium: legumes,
BGA-Azolla: Rice,
Targeted All crops Azotobacter, Azospirillum :
Most cereals, cotton,
sugarcane
Shelf life long Short for bacteria, long for
BGA
Accessibility Affordable section Small and marginal farmers
Irrigation More useful to irrigated field Useful for both irrigated and
dry land farming
Cost High cost input Rs. 6.0 for 1 Low cost input- 20 paise per
kg N, Rs. 14.0 for one kg kg N through Rhizobium, 50
F2O5 paise through BGA
Soil health Indiscriminate use Improves the soil health
deteriorates the soil health
Algal Biofertilizers
Nitrogen fixers

Non-symbiotic

Blue Green Algae FUNGAL BIOFERTILIZER

Phosphate solubilizers

Symbiotic Non-symbiotic
Mycorrhizae Aspergillus
Penicilum
Azotobacter

• Is free living nitrogen fixer

Microscopic observation
• Isolation is carrried out from soil
near rhizosphere area.
• Is Gram negative cocobacilli,capsule
forming,motile,cyst former.
• Forms soft, flat,milky white mucoid colonies on
Agar medium
Azotobacter colonies are often slimy,
due to synthesis of
exopolysaccharide, and
pigmented. A wet mount of an
isolated colony off the plate at 1000X
magnification. Note the large size of
the cells. Both cysts (phase bright
ovals) and vegetative cells (phase
dark bacilli) are visible.
Azotobacter colonies on N-free medium

Capsule formation by
Azotobacter chroococcum
Contributing plant microorganisms Crops benefited
nutrient
A) Nitrogen Symbiotic microbe: Pulse legume:
Rhizobium chickpea,lentil,moong,
bean,cowpea
Oil Legume:
Soybean,groundnut

Azolla Rice
Azospirillum Sorghum,Rice,Wheat,
maize,tomato,chilly
Azotobacter Vegetables:
onion,bringal,tomato,ca
bbage
Cereals:wheat,rice,maiz
e,sorghum,
sugarcane
Phosphate
• Phosphorous, the master key element is known to be involved in
functions in the plant growth & development, photosynthesis,
breakdown of sugar, energy & nutrient transfer within the plant &
expression. Phosphorous nutrition benefits plant by producing
deeper & abundant roots. So supply of this element to plant is
essential for achieving optimum crop yield.
• It is supplied through phosphatic fertilizers, animal manures,
phosphate solubilizing microorganisms.
• The P content in average soil is about 0.05%(W/W).The variation
being largely due to differences in weathering intensity & parent
material composition.
• But only 0.1% of the total P is availble to plants because of its low
solubility & its fixation in soil
Thus soil is rich in phosphorous which needs to
be harnessed.
Phosphate Fertilizers

• There are chemical fertilizers which are providing phosphate. These

fertilizers are routinely applied to promote crop yields.
• The phosphate in these fertilizers is initially available to the plant
but it rapidly reacts with soil & becomes progressively less availble
for plant uptake.This is known as chemical fixation of phosphorous.
So now phosphate solubilizing micro-organisms(PSM) dissolving
interlocked phosphates appear to have an importance.

Many fungi, bacteria are potential solubilizers of

bound phosphates in soil
Mechanism of phosphate solubilization
1) Solubilization by production of organic acid
2) Solubilization by phosphatase

Production of acid:
• The major microbiological means by which insoluble phosphorus compound are
mobilized by the production of organic acid which is accompanied by
acidification of medium.
• The organic & inorganic acids convert tricalcium phosphate to the di- &
monobasic phosphates which got available to plants.
• Examples of organic acids – citric acid, fumaric acid, malic acid, lactic acid,
gluconic acid, ketoglutonic acid, glyoxylic acid.

Solubilization by phosphatase:
The liberation of P from organic phosphate compounds is mainly due to the
action of enzymes of esterase type.

Besides these two mechanisms the production of chelating substances H2S,

CO2,mineral acids, siderophores and ion-exchange mechanisms are also
involved in P solubilization by PSMs.
Both free-living and mutualistic species found in this
group. The mutualistic species may be found in root
nodules of legumes where they are important for proper
plant nutrition. Along with the Cyanobacteria these
nitrogen fixing bacteria contain the metabolic pathways
which can "fix" atmospheric nitrogen N2 into reduced
forms (nitrate, nitrite, and ammonia) that higher plants
can use to make amino acids and therefore proteins.
There are 5 genera -- e.g., Rhizobium sp..  The
photograph below shows Rhizobium sp. in the root
nodules of a bean plant.
Need of nitrogen fixation ? ? ? ? ?
• The earth’s atmosphere contains around 78.08%
[N2] nitrogen gas.
• It cannot be used in this form by most living
organisms until it has been fixed, that is reduced
(combined with hydrogen), to ammonia.
• Nitrogen is required by all living organisms for
the synthesis of proteins, nucleic acids and other
nitrogen-containing compounds.
• Biological nitrogen fixation is the reduction of
atmospheric nitrogen to ammonia by micro-
organisms in soil.
Nitrogen Cycle
• The 2 major processes of N2 transformation are:
• Nitrification:
NH4+ → NO3- :
NH4+ →NO2-
NO2- → NO3-
• By bacteria e.g. Nitrobacter, Bacillus, Paracoccus,
Pseudomonas.
• Denitrification:
▫ NO3- → N2
• By bacteria e.g. Azotobacter, Clostridium and
Rhizobium.
Nitrogen Fixation
• N2 is the most stable form of nitrogen and high
energy is required to break the N-N triple bond.
• Therefore only microorganisms can fix nitrogen
• N2 + 8H+ +8e- →2NH3 + H2
• N2 gas is the greatest reserve of nitrogen.
• The productivity of many environs is limited by
the short supply of nitrogenous compounds.
• Nitrogen fixation is important to agriculture
and legumes such as soybean can fix
atmospheric nitrogen.
Denitrification
• Denitrification is the reduction of nitrates to
N2 or NO2.
• This process is detrimental because it removes
nitrogen the environment.
• This is of particular importance to agriculture
where nitrate fertilizers are used.
• If anoxic condition develop e.g. water logged
soil. The nitrate is remove from the soil by
dentrification.
• What do you understand by the term “anoxic conditions”?
Ammonification
• The decomposition of organic nitrogen
compounds such as amino acids and nucleotides
is called ammonification.
• In the soil much of this NH3 is converted to
amino acids by plants.
• Some NH3 is lost by evaporation especially in
dense animal populations.
• Globally this constitutes 15% of N2 released to
the atmosphere.
Nitrification
• Nitrification is the oxidation of NH3 to NO3- by
nitrifying bacteria.
• The nitrates produced is readily assimilated by
plants.
• Nitrate is soluble and is quickly leached from the
soil.
• NH4+ is +vely charged and will adhere to –vely
charged soil (clay) particles.
• NH4+ is extensively used in nitrogenous
fertilizers.
• Denitrification consumes N2 while nitrification
produces it.
Nitrogen Fixation by Legumes
• The association of nitrogen fixing bacteria with
legumes is one of the most important bacteria plant
interaction.
• Nitrogen fixing legumes include, soybean, bean,
pea, clover and alfalfa are plants with beans in
pods.
• Nitrogen fixing bacteria in plants include:
Rhizobium
Bradyrhizobium
Mesorhizobium
Azorhizobium
30000 X magnified image of Rhizobium
Various groups of Rhizobium
Rhizobium spp. Cross inoculation grouping
R. leguminosarum Pea group
R. phaseoli Bean group
R. trifoli Clover group
R. meliloti Alfalfa group
R. Llupini Lupini group
R. japonicum Soyabean group
R. spp Cowpea group

Tricalcium phosphate solubilizing activity

of Rhizobium sp. strain U9709-SC. The
strain was grown on Illmer and Shinner
agar medium supplemented with
Ca3(PO4)2 at 27 C in the dark for 3 days
O

(right) and 12 days (left). Clear zones

around colonies reveal the areas where
the tricalcium phosphate has been
solubilized.
Nitrogen Fixation
• The symbiotic relationship between plant and
nitrogen fixing bacteria results in the formation
of a root nodule.
• In the nodule N2 is converted by the enzyme
nitrogenase to ammonia.
• The ammonia is used in the synthesis of
amino acids and other cellular components.
• Under normal conditions neither Rhizobium nor
the plant can fix nitrogen.
Root
Nodules
Nitrogen Fixation
• Rhizobium can only fix N2 (diazotrophy) under
microaerophilific (reduced O2) conditions.
• This is because O2 is needed by Rhizobium but O2 also
inhibits nitrogenase.
• Rhizobia are soil bacteria that fix nitrogen after becoming
established inside root nodules of legumes (Fabaceae)
• Rhizobia require a plant host; they cannot independently
fix nitrogen.
• Morphologically, they are generally gram negative, motile,
non-sporulating rods.
Leghemoglobin
• In the nodule O2 level is reduced by
leghemoglobin.
• Leghemoglobin is synthesized only after
interaction of the plant and Rhizobium.
• 90% of legumes will fix nitrogen.
• However each nitrogen fixing bacteria will only
associate with certain legumes.
Leghemoglobin
Steps in Nodule Formation
1. Recognition of the correct partner by both plant
and bacteria.
2. Attachment of the bacteria to the plant root.
3. Invasion of the root hair by bacteria through the
formation of an infection thread.
4. Growth to the main root via the infection thread.
5. Formation of bacteroids (deformed bacteria cells)
and development of nitrogen fixing state.
6. Continued division of plant and bacteria cell and
formation of mature root nodule.
Steps in Formation of Root Nodule
Steps in Formation of Root Nodule
Problems with Biofertilizers
• Microbial fertilizers are supplementary to
chemical fertilizers but not substitute to it.
Microbial fertilizers, usually cause 20 to 30 per
cent increase inn crop production. They do not
cause marked increase in productivity like
chemical fertilizer.
• Specific fertilizer are to be used for specific
crops. This is more applicable to symbiotic
microorganisms. If non specific Rhizobium is
used as fertilizer they do not cause rood
nodulation and increase in crop production.
Problems with Biofertilizers
• Strict aseptic precaution is required during
production of microbial fertilizer. Contamination
is a common problem during microbial mass
production.
• Microbial fertilizer are sensitive to sunlight
exposure. They get killed if exposed for long time
in sunlight.
• Microbial fertilizer must be used within six
months after production when stored at room
temperature. They can be used within two years
if stored at chilling temperature.
Problems with Biofertilizers
• Efficiency of microbial fertilizer is markedly
dependent on soil character e.g. moisture
content, pH, temperature organic matter and
types of resident micro-organisms. When these
factors are unfavorable microbial fertilizer may
not be effective in increasing the soil fertility.
Thank You