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Bio-fertilizers- A Gateway to Sustainable Agriculture

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 Issue

Popular Kheti Special

on
Volume -1, Issue-4 (October-December), 2013 Organic
Available online at www.popularkheti.info Farming
© 2013 popularkheti.info
ISSN:2321-0001 1(4) 2013
Bio-
Bio-fertilizers- A Gateway to Sustainable Agriculture
Balaram Mohapatra1*, Deepak Kumar Verma2, Anindita Sen3, Bipin Bihari Panda4 and Bavita Asthir5
1Environmental Molecular Microbiology Lab, Department of Biotechnology,

Indian Institute of Technology (IIT), Kharagpur (West-Bengal) -721302, India

2Department of Agricultural and Food Engineering,

Indian Institute of Technology (IIT), Kharagpur (West-Bengal) -721302, India

3Department of Biotechnology, Haldia Institute of Technology (HIT),

Haldia (West-Bengal) - 721651, India

4Department of Soil Science and Agronomy, Central Rice Research Institute (CRRI),

Cuttack (Odisha) -753006, India

5Department of Biochemistry, Punjab Agricultural University (PAU),

Ludhiana (Punjab) – 141004, India

*Email of corresponding author:
author: balarammohapatra09@gmail.com

Bio-fertilizers are one of the best modern tools and gift of our agricultural science as a
replacement to our conventional fertilizers. Conventional fertilizers contain compost;
household wastes and green manure and are not as effective as chemical fertilizers. So,
farmers often try to use chemical fertilizers in the field for crop development. But
obviously the chemical fertilizers are not environment friendly because of their
chemical toxicity that can cause water, air and soil pollution and can spread cancer
causing agents. Moreover, they may destroy the fertility of the soil in a long run.
Scientists have developed the way of organic farming by use of “Bio-fertilizers” to
prevent chemical pollution in farm lands. Bio-fertilizer contains microorganisms which
promote the adequate supply of nutrients to the host plants and ensure their proper
development of growth and regulation in their physiology. Living microorganisms are
used in the preparation of Bio-fertilizers which have specific functions to enhance
plant growth and reproduction. Bio-fertilizer being essential components of organic
farming play vital role in maintaining long term soil fertility and sustainability.

Introduction
In the past few decades, environmental pollution has become one of the world's major concerns,
as a number of toxic compounds, originating mostly from industrial and agricultural activities
which have effects of bioaccumulation and bio magnifications. Early detection and replacement of

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toxic chemical compounds in the environment, particularly in water and soil and their biological
effects on organisms has therefore become increasingly important. Past trends in conventional
Indian agriculture, including monoculture without crop rotation, overuse of inorganic fertilizers,
and wide-scale applications of broad-spectrum organophosphate pesticides, have hindered the role
of naturally occurring microorganisms. Onset of green revolution and the increasing use of
chemical fertilizers in agriculture could make the country self-dependent in food production but
on the contrary it deteriorated the environment and caused harmful impacts on living beings. The
excess use of chemical fertilizers in agriculture is costly and also have various adverse effects on
soils i.e. depletes water holding capacity, soil fertility and disparity in soil nutrients. Hence, the
need to develop some low cost effective and eco-friendly fertilizers which would work without
disturbing the nature arose. Now, certain species of micro-organisms are being widely used which
have the unique ability to provide natural products that could serve as a good substitute for
chemical fertilizers.

What are Bio-

Bio-fertilizers?
A live formulation of micro-organisms (bacteria, fungi and algae) are used for fertilization of farm
lands in the aspect of fixation of N2, solubilization and mobilization of phosphorus, increasing
organic carbon content, balanced C/N ratio, plant growth promotion by increasing nutrient
absorption, antagonistic activity against plant pathogens, production of hormones etc that are
beneficial for agriculture. The Gazette of India (2006) defined bio-fertilizer as a product
containing carrier based (solid or liquid) living micro-organisms that are agriculturally useful in
terms of nitrogen fixation, phosphorous solubilization or nutrient mobilization. Bio-fertilizers
were promoted through integrated plant nutrient systems (IPNS) that involved combining
fertilizers, organic/green manures and bio-fertilizers to sustain crop production, maintaining soil
productivity, health and diversity.

Why Bio-
Bio-fertilizers?
As a safe alternative to chemical fertilizer, it minimizes the ecological disturbance and are cost
effective, eco-friendly and when they are required in bulk can be generated at the farm itself.
They increase crop yield upto 10-40% and fix nitrogen upto 40-50 Kg and parental inoculums are
sufficient for growth and multiplication of organisms in subsequent years. They improve soil
texture, pH, WHC and other properties of soil. They produce plant growth promoting substances
IAA, amino acids, vitamins etc. Bio-fertilizers contained 3.5% - 4% nitrogen, 2% - 2.5%
phosphorus and 1.5% potassium. In terms of N: P: K, it was found to be superior to farmyard

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manure and other type of manure. Microbes (Table. 1) are effective in inducing plant growth,
enhance seed germination and root and shoot growth, decomposition of organic materials and
enrichment of compost.
Table. 1. Microbes used as Bio-
Bio-fertilizers
Groups Examples
N2 Fixing Bio
Bio--fertilizers
Free-living Beijerinkia, Azotobacter,Anabaena,Nostoc,
Symbiotic Rhizobium, Frankia,Anabaena azollae
Associative Symbiotic Azospirillum
P Solubilizing Bio-
Bio-fertilizers
Bacillus megaterium,Bacillus subtilis, Bacillus circulans,
Bacteria
Pseudomonas striata
Fungi Penicilliumspp, Aspergillusawamori
P Mobilizing Bio-
Bio-fertilizers
Arbuscularmycorrhiza Glomus spp., Gigaspora spp., Acaulospora spp.
Ectomycorrhiza Laccaria spp., Pisolithus sp., Boletus sp., Amanita spp.
Ericoid mycorrhizae Pezizellaericae
Orchid mycorrhiza Rhizoctoniasolani
Bio-
Bio-fertilizers for Micro nutrients
Silicate and Zinc solubilizers Bacillus spp.
Plant Growth Promoting Rhizobacteria
Pseudomonas Pseudomonas fluorescens
Source:http://www.agritech.tnau.ac.in/org_farm/orgfarm_biofertilizertechnology

Types of Bio-
Bio-fertilizers Available in Market
1. For Nitrogen:
1. Rhizobium for legume crops.
2. Azotobacter/Azospirillum for non-legume crops.
3. Blue Green Algae (BGA) and Azolla for low land paddy.
2. For Phosphorous:
1. Phosphatika for all crops to be applied with Rhizobium, Azotobacter, Azospirillum
and Acetobacter
3. For Enriched Compost:
1. Cellulolytic fungal culture
2. Phosphotika and Azotobacterculture

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Microorganisms Used as Bio-

Bio-fertilizers
1. Nitrogen Fixing Bacteria
Rhizobia: Nitrogen, as a principal component of cell biomolecules (Amino acid, proteins,
enzymes), all plants need it as a sole source for their metabolism. Mostly leguminous plants
require high amount due to their protein demand and can’t fix atmospheric N2 due to high
energy expenditure. So, Legumes plants have root nodules, where atmospheric nitrogen fixation
is done by bacteria belonging to genera, Rhizobium, Bradyrhizobium, Sinorhizobium,
Azorhizobium and Mesorhizobium collectively called as rhizobia, belong to α-Proteobacteria.
When rhizobial culture is inoculated in field, pulse crops yield can be increased due to rhizobial
symbiosis (Dubey, 2001). Rhizobium can fix 15-20 N/ha and increase crop yield upto 20% in
pulses.
Table.2. Rhizobium-
Rhizobium-plant Associations
Rhizobium Host Plant
R. meliloti Medicago, Melilotus, Trigonellaspp.
R. leguminosarum
bv. Viciae Pisum, Vicia, Lathyrus, Lens spp.
bv. Trifolii Trifoliumspp.
bv. Phaseoli Phaseolus vulgaris
R. ciceri Cicerarietinum
R. tropici Phaseolus vulgaris, Leucaenaspp., Macroptiliumspp.
R. etli Phaseolus vulgaris
R. galegae Galegaofficinalis, G. Orientalis
R. fredii Glycine max, G. soja, and other legumes
B. japonicum Glycine max, G. soja, and other legumes
B. elkanii Glycine max, G. soja, and other legumes
A. caulinodans Sesbaniaspp. (stem nodulating)
Source: vanRhijnand Vanderleyden (1995)
Azorhizobium: It is a stem nodulating and nitrogen fixing symbionts of the stem nodule, it also
produce large amount of IAA that promotes plant growth.
Bradyrhizobium: Bradyrhizobium is a good nitrogen fixer. Bradyrhizobium strain inoculation
with Mucuna seeds enhances total organic carbon, N2, phosphorus and potassium in the soil,
increases plant growth and consequently plant biomass, reduction in the weed population and
increased soil microbial population.

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2. Diazotrophs
Azotobacteracae (e.g. Azotobacter)
Azotobacter): They are the free living, non-symbiotic, aerobic,
photoautotrophic bacteria. They secretes vitamins, gibberellins, naphthalene, acetic acid and
other substances that inhibit certain root pathogens and improves root growth and uptake of
plant nutrients. It occurs in the roots of Paspalumnotatum (tropical grasses) and other spp. and
adds 15-93 Kg N/ha/annum on P. notatum roots. Azotobacter indicum occurs in acidic soil in
sugarcane plant roots. It can apply in cereals, millets, vegetables and flowers through seed,
seedlings soil treatment.
Spirillaceae (e.g. Azospirillum and Herbaspirillum)
Herbaspirillum): These are gram negative, free living,
associative symbiotic and non-nodule forming, aerobic bacteria, occurs in the roots of dicots and
monocot plants i.e. corn, sorghum, wheat etc. It is easy to culture and identify. Azospirillum is
found to be very effective in increasing 10-15% yield of cereal crops and fixes N2 upto 20-40%
Kg/ha. Different A. brasiliense strains inoculation in the wheat seed causes increase in seed
germination, plant growth, plumule and radicle length. Herbaspirillum species occurs in roots,
stems and leaves of sugarcane and rice. They produce growth promoters (IAA, Gibberillins,
Cytokinins) and enhance root development and uptake of plant nutrients (N, P & K).
Acetobacterdiazotrophicus: Another diazotroph is Acetobacterdiazotrophicus occurs in roots,
stem and leaves of sugarcane and sugar beet crops as nitrogen fixer and applied through soil
treatment. It also produces growth promoters e.g. IAA and helps in nutrients uptake, seed
germination, and root growth. This bacterium fixes nitrogen upto 15kg /ha/year and enhance
upto 0.5 – 1% crop yield (Gahukar, 2005-06).
Cyanobacteria (Blue green algae): Nostoc, Anabaena, Oscillatoria, Aulosira, Lyngbya etc. are the
prokaryotic organisms and phototropic in nature. They play an important role in enriching paddy
field soil by fixing atmospheric nitrogen and supply vitamin B complex and growth promoting
substance which makes the plant grow vigorously. Cyanobacteria fixes 20-30 Kg/N/ha and
increase 10-15% crop yield when applied at 10 Kg/ha. Both free-living as well as symbiotic
cyanobacteria (blue green algae) used in rice cultivation in India.
Azolla – Anabaena symbiosis:
symbiosis: It is a free floating, aquatic fern found on water surface having a
cyanobacterial symbiont Anabaena azollae in their leaves. It fixes atmospheric nitrogen in paddy
field and excrete organic nitrogen in water during its growth and also immediately upon
trampling. Azolla contributes nitrogen, phosphorus (15-20 Kg/ha/month), potassium (20-25
kg/ha/month) and organic carbon etc. and increases 10-20% yield of paddy crops and also
suppresses weed growth. Azolla also absorbs traces of potassium from irrigation water and can be

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used as green manure before rice planting. Azolla spp. are metal tolerant hence, can be applied
near heavy metal polluted areas.
3. Phosphate Solubilising Bacteria:
Bacteria: Solubilization of inorganic tricalcium and rock phosphate
occurs by most of bacteria belong to genus: Pseudomonas, Bacillus, Acrobacter, Nitrobacter,
Escherichia, Serratia spp. and speciallyPseudomonas striata, Bacillus polymyxa are the bacteria
have phosphate solubilising ability. ‘Phosphobacterin’ are the bacterial fertilizers containing cells
of Bacillus megatherium var. phosphaticum, prepared firstly by USSR scientists. They increased
about 10 to 20 % crop yield and also produces plant growth promoting hormones which helps in
phosphate solubilising activity of soil.
4. Phosphate Solubilizing Fungi:
Fungi: Some fungi also have phosphate dissolving ability e.g.
Aspergillus niger, Aspergillus awamori, Penicillium digitatum etc. All the microbes solubilize
phosphate by production of organic acids.
5. Silicate Solubilizing Bacteria:
Bacteria: Microorganisms are capable of degrading silicates and aluminum
silicates. During the metabolism of microbes several organic acids are produced and these have a
dual role in silicate weathering. They supply H+ ions to the medium and promote hydrolysis and
the organic acids like citric, oxalic acid, Keto acids and hydroxy carbolic acids which from
complexes with cations, promote their removal and retention in the medium in a dissolved state.
Most of soil born Bacillus spp. can be used as silicates solubilizers.
6: Plant Growth Promoting Rhizobacteria (PGPR):
(PGPR): The group of bacteria that colonize roots or
rhizosphere soil and beneficial to crops are referred to as plant growth promoting rhizobacteria
(PGPR). They are also called as microbial pesticides e.g. Bacillus spp. and Pseudomonas
fluorescence. Serratia spp. and Ochrobactrum spp. are able to promote growth of plants. The
PGPR inoculants currently commercialized that seem to promote growth through at least one
mechanism; suppression of plant disease (termed Bio-protectants), improved nutrient acquisition
(termed Bio-fertilizers), or phyto-hormone production (termed Bio-stimulants). Species
of Pseudomonas and Bacillus can produce as yet not well characterized phyto-hormones or
growth regulators that cause crops to have greater amounts of fine roots which have the effect of
increasing the absorptive surface of plant roots for uptake of water and nutrients. These PGPR
are referred to as Bio-stimulants and the phyto-hormones they produce include indole-acetic
acid, cytokinins, gibberellins and inhibitors of ethylene production.
7. Mycorrhiza:
Mycorrhiza: Mycorrhizas are symbiosis between some specific root inhabiting fungi and plant
roots and used as Bio-fertilizers. They absorb nutrients such as manganese, phosphorus, iron,

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sulphur, zinc etc. from the soil and pass it to the plant. Mycorrhizal fungus increases the yield of
crops by 30-40% and also produces plant growth promoting substances.
8. VAM fungi or Endomycorrhiza:
Endomycorrhiza: They form arbuscles in the roots of crop plants and enhance
nutrient uptake: phosphorus and other nutrients that are responsible for plant growth
stimulation including roots and shoot length. VAM also enhances the growth of black pepper and
protects from Phytophthora capsici, Radopholus similis and Melvidogyne incognita (Anandraj et
al., 2001). VAM fungi enhance water uptake in plants and also provide heavy metals tolerance to
plants..
Application of Bio-
Bio-fertilizers
Seed treatment:
treatment: One packet of the inoculants (200 g) is mixed with 200 ml of rice kanji to make
slurry. The seeds required for an acre are mixed in the slurry so as to have a uniform coating of
the inoculant over the seeds and then shade dried for 30 minutes. The shade dried seeds should
be sown within 24 hours. One packet of the inoculant (200 g) is sufficient to treat 10 kg of seeds.
Seedling root Dip:
Dip: It has better application for transplanted crops. Two packets of the inoculant
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:
application: 1kg of the inoculant is mixed with 20 kg of dried and powdered farm yard
manure and then broadcasted in one acre of main field just before transplanting.
Mass Production of Bio-
Bio-fertilizers
The mass production of Bio-fertilizers involves three stages:
Stage-
Stage-1: Culturing of microorganisms
Stage-
Stage-2: Processing of carrier material
Stage-
Stage-3: Mixing the carrier and the broth culture and packing
Bio-fertilizers are carrier based preparations containing efficient strain of nitrogen fixing or
phosphate solubilizing microorganism prepared in suitable medium and mass produced in
fermentor with proper aeration, temperature, growth conditions and without any undesired
microbial contaminations. Bio-fertilizers are formulated usually as carrier based inoculants. The
organic carrier materials are more effective for the preparation of bacterial inoculants. Peat soil,
lignite, vermiculite, charcoal, press mud, farmyard manure and soil mixture can be used as carrier
materials. The neutralized peat soil/lignite are found to be better carrier materials for bio-
fertilizer production which is cheaper, locally available, inert, having high WHC and organic
content. The bacterial culture drawn from the fermentor is added to the sterilized carrier and
mixed well by manual (by wearing sterile gloves) or by mechanical mixer then sealed at room
temperature.

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Table 3. Microorganisms as Bio-

Bio-fertilizers for
for different crops
Microorganism Nutrient fixed Host Crops for
(Kg/ha/year)1 which Used
Actinorrhizae 150 kg N/ha For certain non-legumes mainly
(Frankia spp.) trees & Shrubs
Algae 25 kg N/ha Rice
Azolla 900 kg N/ha Rice
Azospirillum 50 to 300 kg N/ ha Non-legumes like maize, barley,
oats, sorghum, millets sugarcane,
rice etc
Rhizobium 0.026 to 20 kg N / ha Legumes like pulses, peas,
Groundnut, soybean, beans, and
clover
Azotobacter 10-20 kg N /ha Cereals, millets, cotton, vegetables
Mycorrhizae (VAM) Solubilize food phosphorus (60%) Many tree species, wheat, sorghum,
ornamentals
Phosphate solubilizing Solubilize about 50-60% of them Soil application for all crops
Bacteria and fungi fixed phosphorus in the soil
Sources: Mall et al., (2013)
Table.4. Recommended liquid bio- bio-fertilizers and its application method, quantity to be used for
different crops are as follows:
follows
Recommended Application Quantity to be
Bio-
Bio-fertilizer method used Crops
Rhizobium Seed treatment 200ml/acre Chickpea, pea, Groundnut,
soybean, beans, Lentil, Green
gram, Black gram, Cowpea and
pigeon pea
Azotobacter/Azospirillum Seed treatment 200ml/acre Wheat, oat, barley
Azospirillum Seed treatment 200ml/acre Rice
Azotobacter Seed treatment 200ml/acre Mustard, seasum, Linseeds,
Sunflower, castor
Azotobacter Seed treatment 200ml/acre Pearl millets, Finger millets, kodo
millet
Azospirillum Seed treatment 200ml/acre Maize and Sorghum
Azotobacter Seed treatment 200ml/acre Forage crops and Grasses
Azotobacter Soil treatment 400ml/acre Tea, Coffee
Azotobacter Soil treatment 2-3 ml/plant Rubber, Coconuts
Rhizobium Soil treatment 1-2 ml/plant Leguminous plants/ trees
Note: Doses recommended when count of inoculum is 1 x 108 cells/ml then doses will be ten times more besides
above said Nitrogen fixers, Phosphate solubilizers and potash mobilizers at the rate of 200 ml/ acre could be applied
for all crops.

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Storage of Bio-
Bio-Fertilizer Packets
• The packet should be stored in a cool place away from the heat or direct sunlight.
• The packets may be stored at room temperature or in cold storage conditions in lots in
polythene /gunny bags.
• The population of inoculant in the carrier inoculant packet may be determined at 15 days
interval. There should be more than 109 cells / g of inoculant at the time of preparation.
Constraints in Bio-
Bio-fertilizer Technology
In spite of low cost, eco-friendliness, several constraints limit the application or implementation
of the technology. The constraints may be environmental, technological, infrastructural,
financial, human resources, unawareness, quality, marketing, etc.
Technological constraints:
onstraints: Use of improper, less efficient strains for production, lack of qualified
technical personnel, good quality carrier material, production of poor quality inoculants without
understanding the basic microbiological techniques and short shelf life of inoculants.
Infrastructural constraints:
onstraints: Lack of essential equipments, non-availability of suitable facilities for
production, space availability for laboratory, production, storage etc.
Financial constraints:
onstraints: Non-availability of sufficient funds and problems in getting bank loans and
less return by sale of products in smaller production units.
Environmental
Environmental constraints:
onstraints: Seasonal demand for Bio-fertilizers, simultaneous cropping
operations and short span of sowing/planting in a particular locality, soil characteristics like
salinity, acidity, drought, water logging, etc.

Conclusion
As a boon for farmers, Bio-fertilizers being essential components of organic farming play vital
role in maintaining long term soil fertility and sustainability. Bio-fertilizers would be the viable
option for farmers to increase productivity per unit area in organic farming for an era of
prosperity and clean environment.

References
Anandraj M, Venugopal MN, Veena SS, Kumar A and Sarma YR. 2001.Ecofriendly management of
disease of species.Indian species. 38 (3): 28-31.
Dubey RC. 2001. A text book of biotechnology. S. Chand & Company Ltd. New Delhi.
Gahukar RT 2005-06. Potential and use of bio-fertilizers in India.Evermans science. XL (5): 354-361.
Mall RK, Verma DK, Tripathi HC, Pathak RK and Asthir B. 2013. Bio-fertilizers in Context of
Farmers and Agriculture in India.Indian Farmer’s Digest.46 (1): 16-18.
vanRhijn P and Vanderleyden J. 1995. The Rhizobium-Plant Symbiosis.Microbiology Review.59 (1):
124–142.

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