Submitted by:

Anusha Aditya (4th sem)

Rollno:1060015
 Bio fertilizer

Bio fertilizers are not fertilizers. “Bio fertilizer” is a substance

which contains living microorganisms which, when applied to
seed, plant surfaces, or soil, colonizes the rhizosphere or the
interior of the plant and promotes growth by increasing the
supply or availability of primary nutrients to the host plant.

Why bio fertilizers?

It’s a microbial green revolution. Bio fertilizers are having it

sown advantages over chemical fertilizers and it is
economically and environmental friendly too. With the
increasing demand in agriculture it has become important for
us to increase the Productivity by using various fertilizers
insecticides Pesticides .But with the tremendous use of these
products the soil has been affected badly because of the
depletion in the essential minerals of the soil. So to overcome
this problem it has become important for all of us to use a
different remedy for the production of various bio fertilizers.
They are the best at economic value.

Types of bio fertilizers:

 Nitrogen bio fertilizers

 Compost bio fertilizers
 Phosphorous bio fertilizers
 Nitrogen fixing fertilizers:

Symbiotic: Rhizobium

It belongs to rhizobiaceae family, the rhizobium bacteria

present in the nodules of these crops are not always efficient.
Therefore, the competitive, efficient bacteria are isolated,
screened, selected and produced as carrier based inoculants.

Morphology:

1) Unicellular, cell size less than 2µ wide. Short to medium rod,

pleomorphic
2) Motile with peritricus flagella
3) Gram negative
4) Accumulate poly β-hydroxyl butyrate granules.

Physiology:

1) Nature : chemo heterotrophic, symbiotic with legume

2) C source: supplied by legume through
photosynthesis, mono & disaccharide.
3) N source: fixed from atmosphere.
4) Respiration: aerobic.
5) Growth: fast ( rhizobium), slow (Brady rhizobium)
6) Doubling time: fast grower- 2-4 hours slow grower 6-12 hours.
7) Growth media : YEMA
 Recommended for :

Pulses: chickpea, pea, lentil, black gram, green gram, cowpea,

pigeon pea.

Oil seeds: soybean, groundnuts.

Non symbiotic: Azospirillum, Azotobacter.

Azotobacter

It belongs to azotobacteriaceae .It produces growth promoting

substances which improve seed germination and growth of
extended root system. It produces polysaccharides which
improve soil aggregation. Azotobacter suppresses the growth of
saprophytic and pathogenic micro-organism near the root
system of crop plants.

Morphology:

1) Cell size: Large ovoid cells, size ranging from 2.0-7.0×1.0-2.5µ.

2) Cell character: polymorphic
3) Accumulate poly β-hydroxyl butyrate granules.
4) Gram reaction: negative.
Physiology:

1) Nature: chemo heterotrophic, free living

2) C source: a variety of carbon source ( mono, di
and certain polysaccharide) organic acids.
3) N sources: Nitrogen through fixation, amino acid, NH4, NO3
4) Respiration: aerobic
5) Growth media: Ashby Jensen’s medium
6) Doubling time: 3 hours.

Contribution :

1) 20-40 mg BNF/g of C source in laboratory condition equivalent

to 20-40 kg N/ha.
2) Production of growth promoting substance like
vitamins of B groups, indoleacetic acid and gibberellin acid.
3) Biological control of plant disease by suppressing Aspergillus,
Fusarium.

Recommended for:

Rice, wheat, millets, other cereals, cotton, vegetable, sunflower,

mustard, flowers.

Increase in yield: 20 to 30%

Azospirillium

It belongs to family spirillaceae. The bacteria have been found to live

within the root of sorghum, bajra and rage plants. They are
chemoheterotrophic and association in nature secrete growth
regulatory substance
The use of azospirillium inoculants help in increasing yield of millets. It
significantly increase the growth, chlorophyll content and mycorrhyzal
infection in root. Increased growth and nutrient uptake by barley plants
were observed when seed were co-inoculated with A. baselines and
Glomus vermiform.

Morphology:

1) Cell size: curved rod, 1mm in diameter, size and shape vary.
2) Accumulate: poly β-hydroxyl butyric acid.
3) Gram reaction: negative
4) Development of white pellicles 2-4mm below the surface of NFB
medium.
Physiology:

1) Nature: chemoheterotrophic, associative.

2) C source: organic acid, L-
arabinose,
D-gluconate, D-fructose.
3) N sources: nitrogen through fixation, amino acids, NH4, NO3
4) Respiration: aerobic, micro aerobic.
5) Growth media: N free bromothymol blue (NBF)
6) Doubling time: 1hr in ammonia containing
medium, 5.5 to 7hr on malate containing semi-
solid medium.
 Contribution:

1) 20-40 mg N/g malate under laboratory condition equivalent to 20-40 kg

N/ha.
2) Results in increase mineral and water uptake, root
development, vegetative growth and crop yield.

Recommended for: Rice, millets maize, wheat, sorghum, sugarcane and

co-inoculants.

Response: Average increase in yield : 15-30%.

Phosphate solubilizing bio fertilizer

Phosphorus is one of the most important plant nutrients and may be

critical nutrient for the optimum growth of plants. Most of our soils are
in available forms of phosphorus required phosphate application.
In the rhizosphere of crops will render insoluble soil phosphate
available to plants due to production and secretion of organic acid
by them. The use of this bio fertilizer will also increase the
availability of phosphate from rock phosphate applied directly
even to neutral to alkaline soil or when used for preparation of
phosphor-compost. Phosphate solubilizing micro-organism
include efficient strain of bacteria, fungi, yeast and actinomycetes
in that order

Bacteria Morphology:

1) Cell size: rod shape, 1.1 to 2.2µm in diameter.

2) Gram reaction: for Bacillus positive and for Pseudomonas
negative
3) Transparent zones of clearing around microbial colonies
indicate extent of Phosphate solubilization.

Physiology:
1.)Nature: chemoheterotrophic.
2.) C source: Glucose is the main C source but
they can utilize other carbon sources.
 ADVANTAGES OF BIOFERTILIZERS:-

1) They help to get high yield of crops by making the soil rich
with nutrients and useful microorganisms necessary for the
growth of the plants.

2) Bio fertilizers have replaced the chemical fertilizers as

chemical fertilizers are not beneficial for the plants. They
decrease the growth of the plants and make the environment
polluted by releasing harmful chemicals.

3) Plant growth can be increased if bio fertilizers are used, because

they contain natural components which do not harm the plants but
do the vice versa.

4) If the soil will be free of chemicals, it will retain its fertility which
will be beneficial for the plants as well as the environment, because
plants will be protected from getting any diseases and environment
will be free of pollutants.
5) Bio fertilizers destroy those harmful components from the soil
which cause diseases in the plants. Plants can also be protected
against drought and other strict conditions by using bio fertilizers.

6) Bio fertilizers are not costly and even poor farmers can make use of them.

7) They are environment friendly and protect the environment against

pollutants.

Disadvantages

 Much lower nutrient density -- requires large amounts to get enough

for most
crops.
 Requires a different type of machine to apply than chemical fertilizers.
 Sometimes hard to locate in certain areas odor.
 Why do we need bio fertilizers……?????????????

An estimate shows 100million tons of fixed N2 is required

for global food production. Chemical fertilizers is the most
common practice to increase crop yields Besides the cost
factor the use of chemical fertilizers is associated with
environmental pollution.
\
 Process of making bio fertilizer:

• Bio fertilizers are usually prepared as carrier-based

inoculants containing effective microorganism.

• Incorporation of microorganisms in carrier material

enables easy-handling, long-term storage and high
effectiveness of bio fertilizers.

• Among various types of bio fertilizers, bacterial

inoculant is one major group which includes rhizobia,
nitrogen-fixing rhizobacteria, plant growth
promoting
rhizobacteria, phosphate-solubilizing bacteria.

• Basically, the carrier-based inoculant of these bacteria can

be prepared by a common procedure.
 The most common way of inoculation

Seed inoculation

The inoculant(bacteria-carrier mixture) is mixed with water to

make slurry- form, and then mixed with seeds. In this case, the
carrier must be a form of fine powder. To achieve the tight coating
of inoculant on seed surface, use of adhesive, such as gum arabic,
methylethylcellulose, sucrose solutions, and vegetable oils, is
recommended. Seed inoculation may not always be successful, i.e.
the inoculation resulted in low nodule. occupancy of the inoculated
rhizobial strain, or low establishment of the inoculated
rhizobacterial strain. This might be due to low population and/or
low survival of the inoculated bacterial strain on the seed surface
and in the soil.

“soil inoculation” will be adopted, whereby a large population of a

bacterial strain can be introduced into the soil. For soil inoculation
in general, granular inoculant is placed into the furrow under or
alongside the seed. This enhances the chance for the inoculated
strain to be in contact with plant roots.
 Phosphate solubilizing culture

Rhizobium culture
 Azospirillum
culture

Azotobacter and Rhizobium

Carrier material:

Various types of material are used as carrier for seed or soil

inoculation. For preparation of seed inoculant, the carrier
material is milled to fine powder with particle size of 10 -40
μm.

The properties of a good carrier material for seed

inoculation are:
(1) Non-toxic to inoculant bacterial strain.
(2) Good moisture absorption capacity.
(3) Easy to process and free of lump-forming materials.
(4) Easy to sterilize by autoclaving or gamma-irradiation.
(5) Available in adequate amounts
(6) Inexpensive.
(7) Good adhesion to seeds, and
(8) Good pH buffering capacity.
(9) Need less non-toxic to plant, is another important property.

Essential criteria for carrier selection relating to

survival of the inoculant bacteria should
be considered :

(1) Survival of the inoculant bacteria on seed. Seeds are not

always sown immediately after seed coating with the
inoculant bacteria. The bacteria have to survive on seed
surface against drying condition until placed into soil.
(2) Survival of the inoculant bacteria during the storage period.
(3) Survival of the inoculant bacteria in soil.
 After being introduced into the soil, the inoculants bacteria
have to compete with native soil microorganisms for the
nutrient and habitable niche, and have to survive against
grazing protozoa. Such carrier materials that offer the available
nutrient and/or habitable micro-pore to the
inoculants bacteria will be desirable. In this sense, materials with
micro-porous structure, such as soil aggregate and charcoal, will
be good carrier for soil inoculants.

Sterilizations:

Carrier sterilization is autoclaving. Carrier material is packed in

partially opened, thin-walled polypropylene bags and autoclaved

for 60 min at 121 ºC. It should be noted that during

autoclaving, some materials
changes their properties and
produce toxic substance to
some bacterial strains.
Inoculant Parameters Sterile Non-sterile

Population of beneficial High Variable

bacteria

Choice of materials to be Many materials are not easily Almost unlim

used as carriers sterilized or change their
chemical and physical
composition upon sterilization

Labor requirements Skilled & expensive Mostly unsk

Sterile production space Large and costly Not needed

Monitoring of contamination Essential for quality Essential for

control of the product product

Total cost of production High Much lower

Longevity High Relatively lo

Sterilization equipment Huge autoclave machine Not needed

required they are very costly
 New Trends in Formulations Using Unconventional
Synthetic Materials

These polymers have demonstrated potential as bacterial carriers

that offered substantial advantages :

 These formulations encapsulate the living cells protect

the microorganisms against many environmental
stresses, and release them to the soil, gradually but in
large quantities.
 when the polymers are degraded by soil microorganisms,
usually at the time of seed germination and seedling
emergence.
 They can be stored dried at ambient temperatures for
prolonged periods, offer a consistent batch quality and a
better defined environment for the bacteria, and can be
manipulated easily according to the needs of specific
bacteria.
 These inoculants can be amended with nutrients to
improve the short-term survival of the bacteria upon
inoculation.
Encapsulated Formulation
• The encapsulation of microorganisms into a polymer
matrix is still experimental in the field o f bacterial-
inoculation technology.
• At present there is no commercial bacterial product using this
technology.
• The concept underlying immobilized microbial
cells, is to entrap beneficial microorganisms
into a matrix.
• The formulation (bacteria-matrix) is then fermented in a bacterial
growth medium.
The main goal of these industrial formulations is to maintain
the cells entrapped in an active form for as long as possible.
Any premature release of the microorganisms from these
encapsulated forms is undesirable.

Encapsulated bacterial formulations in agriculture have at

least two distinctly different goals from those of the
fermentation industry:

a) To temporarily protect the encapsulated microorganisms from the

soil environment
and microbial competition, and

b) To release them gradually for the colonization of plant roots.

 Alginate

Alginate is the material most commonly used for

encapsulation of microorganisms. The resulting
inoculation are used for various purposes:

 The immobilization of cell organelles and enzymes,

 The application of biological control agents and mycoherbicides

 To increase the stability of recombinant plasmids in the host

cells.
 REGULATION AND CONTROL OF CONTAMINATION OF
COMMERCIAL INOCULANTS

Naturally, an inoculant should contain a level of bacteria

sufficient to inoculate plants and produce an economic gain. The
required level of bacteria cannot be established as a general
standard because it varies from one bacterial species to another.
Only rhizobial inoculants have legally established standards.

Quality control methods to determine the number of

bacteria within the inoculant are not standardized either.
To measure the bacterial number, commonly known
methods in microbiology are used; the traditional Plate
Count methods, Most Probable Number.
 Examples of New Commercial Microbial Inoculants

Commercial microbial inoculants of other beneficial microorganisms have begun to appear

on the market on a small scale. These include "Azogreen", a French-approved Azospirillum
inoculant.

COST OF DEVELOPMENT AND MARKETING

The cost of developing a new product by the agrochemical industry has been estimated at
over $80 million US and rising . The development of resistance to pesticides may shorten the
commercial life of these products and thus their potential return. The development of
bacterial inoculants is claimed to be cheaper than that of agrochemicals.
The following are some factors that reduce the costs of development of bacterial
inoculants which makes them attractive to the agrochemical industry:

(i) Reduced registration costs compared to those of

chemical-product test programs t hat are well- established and costly.

(ii) Reduced registration time decreases the time span from first screening to market,
thus increasing revenues

(iii) The possibility of developing bacterial products for small markets. Since the cost
involved in bringing a new chemical to the marketplace is so large, the product must be
targeted to a market large enough to have a good return
on investment. This limits the choice of crops to the major crops only.

(iv) Although fermentation is costlier than chemical production, the

fermentation plant is more versatile.
Other motivational steps for the agrochemical industry to develop
bacterial inoculants might be:

• It is less likely that pathogens will develop resistance as fast as they do to chemical
products.

• Some bacterial inoculants, especially those that use an organism employing a

single mechanism against the pathogen, can also develop resistance.

• They are "environment friendly". The "natural“ tag of bacterial inoculants (especially
those that are non engineered and indigenous) make them more acceptable in the
public eye, and especially t o the "Green movement" pressure groups, than chemicals.
 Market requirement

First, all the considerations mentioned above (efficient strains, optimized formulations, cost-
effective production, and good and practical inoculation techniques) are not sufficient to launch a
new product on the market nor guarantee its success. The following practical variables should be
considered:

(i) The product must be efficient and reliable in large-scale field trials and especially under "real life"
conditions.

(iii) Obviously, patents on industrial processes and registration of biological products must be
secured

(iv) For every potential customer country, a market survey must be done which examines
customer demand, market size, and expected selling price.
 CONCLUSIONS AND FUTURE PROSPECTS

 The agrochemical industry is more sympathetic now to the concept of bacterial

inoculants than it has been previously. There is a genuine interest in developing
bacterial products that are reliable and that can act as complements to chemicals
already on the market

 Greenhouse crops are also primary targets for commercial inoculants

 Pioneering transgenic plants are already in the field expressing

insecticidal proteins of B. thuringiensis in cotton plants, making them
resistant to various insect pests.

 A gradual and modest increase in the use of bacterial inoculants is to be expected.

 Agriculture in developed countries is definitely the major promoter of microbial

inoculants that are "environmentally friendly“.