BIO-FERTILIZERS

Presented by –
Lakshmi giri
831/18
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 Introduction
 As we know ,with increase in population the demand in agriculture is also
increasing ,it has become important for us to increase the productivity by
using various fertilizers, insecticides ,and 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 us to use a different

remedy for the production of various bio-fertilizers .

 Bio-fertilizers are the substances that contain micro-organisms, which when
added to the soil increases the crop yield and promotes plant growth .
 Bio-fertilizers adds nutrients through the natural processes of nitrogen
fixation ,solubilising phosphorus and stimulating plant growth.

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 WHAT IS BIO FERTILIZER?
• Micro-organisms employed to enhance the availability of
nutrients by nitrogen fixation of atmospheric bacteria and
by solublizing soil phosphorus , to the crops are called bio-
fertilizers .

• Of these , biological nitrogen fixation offers an

economically attractive and ecologically sound route
for augmenting nutrient supply .
• Thus various species of Rhizobium,Blue green algae,
Cynobacteria,Azolla ,Azotobacter ,Mycorrhiza etc ,
plays significant role in agriculture .
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Types of bio-fertilizers
• Symbiotic nitrogen fixers
Bacterial • Free living nitrogen fixers.
• Phophate solublising bacteria

• AMF(arbuscular mycorrhizal fungi)

Fungal

• Blue green algae in association

Algal with azolla
• Anabena,Nostoc,Ocillatoria

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 A .Bacterial fertilizers
 The live cell of bacteria used as a biofertilizers

 These microbes contains unique gene called as Nif-Gene

which makes them capable of fixing nitrogen .
 The nitrogen fixing bacteria works under two condition ,
 Symbiotically
 Free living bacteria (non-symbiotically )
 The symbiotic bacteria make an association with crop
plants through forming nodules in their roots .
 Where as the free living bacteria do not for any association
but live freely and fix atmospheric nitrogen.

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 1.Symbiotic nitrogen fixing bacteria
• Most important symbiotic Nitrogen fixing bacteria is Rhizobium
and Azospirillum .

RHIZOBIUM :-
 These are gram-negative soil bacteria capable of forming root
nodules in most leguminous plants and in some non-leguminous
plants .
 These are micro aerobics .
 In some cases stem nodules are also produced.
 Rhizobium cells contains nif-genes for nitrogen fixation on
megaplasmid .
 It can fix nitrogen from 50 to 150 kg/ha.
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 Organism Activity used in Scientific crop name
crops
1. Azorhizobium N-fixation Acculeata Sesbania rostrata
Caulinodans Nodulates the
stem
2.R.leguminosarum N-fixation Pea Pisum sativum
Nodulates Khesari Lathyrus ativus
lentil Lens culinaris
3 .R.phaseoli N-fixation Beans Phaseolus multiflorus
Nodulates Kidney/french Phaseolus vulgaris
beans
4. R.meliloti N-fixation Lucerne Medicago sativa
Nodulates Fenugreek Trifolium –foenumgraecum

5. R.trifoli N-fixation Clover Trifolium sp.

Nodulates Egyptian clover Trifolium –alexandrinum

6 R.lupini N-fixation White lupines Lupinus alba

Nodulates Lupines Lupinus sp.

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 Contd.
• R.lupines are relatively slow growing ,however R.japonium( the
cowpea rhizobium ) is the slowest growing rhizobial sps.
• The cow pea rhizobium is now known as Bradyrhizobium -
nodulates soyabean (Glycine max.).
• Other species of Rhizobium are known as fast growing Rhizobia.
• Other rhizobial species cow pea miscellany nodulates
sunnhemp,cluster beans ,lablab bans ,pigeonpea etc…
• The National facility for rhizobium culture collections at
IARI,New Delhi serves as a respository for Rhizobium culture .
• The ability to fix nitrogen depend upon both host and bacterial
genes .
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 Mechanism :-
The bacteria enters the root through root hairs ,the interactions being highly specific and progressing
through several steps .

Ultimately results in nodule formation

Inside the nodule many bacterial cells change into non-diving bacterioids ,which produces Nitrogenase

Atmospheric Nitrogen Nitrogenase Amonia

Nitogenase is highly sensitive to oxygen and protected by pink pigments Leghaemoglobin which binds to
oxygen .The pink pigment is produced by the legumes and is present in the nodules .
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 Azospirillum:-
• It is mainly present in grass family like
sugarcane ,jowar,wheat,bajra etc.
• Inhabits both root cells as well as surrounding
of the roots .
• Forms symbiotic relation and increase
Nitrogen –fixing potential of the cereal plant .
• Field inoculation of crops with Azospirillum is
estimated to save 15 to 25 kgN/ha .
• It is commercially exploited for the use as
nitrogen supplying Bio-Fertilizer .
• Culture of these bacteria are conserved at
Microbial type culture collection at institute
of Microbial Technology ,Chandigarh .

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 2.Free living bacteria
• Large number of free living or non-symbiotic
bacteria (does not form nodules but makes
association by living in the
Rhizosphere)present in soil.
• Commonly used free living bacteria are :-
Azotobacter
Klebsiella

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 Azotobactor
• Azotobactor is a heterotrophic free living
nitogen fixing bacteria present in alkaline
and neutral soils.
• It is commonly occurring species in arable
soil of India .
• It uses organic matter present in soil to fix
nitrogen Asymbiotically .
• It is capable of fixing up to 30kgN/ha/yr .
• Apart from N-fixation it also synthesize
growth promoting substances such as
auxins and gibberellins .
• Azotobactor requires large amount of
organic carbon for its growth .

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 B. Algal fertilizers
Blue green algae
• Free living nitrogen fixers Cynobacteria commonly called as blue
green algae .
• These are photosynthetic ,prokaryotic organism ,which fix
nitrogen Asymbiotically .
• Some cynobacteria form symbiotic association
• Nitrogen fixation takes place in specialized cells called Heterocyst
• BGA is very common in fields and unlike azotobactor BGA is not
inhibited by the presence of chemical fertilizers .
• If no chemical fertilizers are added ,inoculation of algae can
result in 10-14%increase in crop yield.
• BGA are beneficial in certain crops like vegetables ,cotton and
sugarcane .
• For example of algal biofertilizers are :-
 Anabena
 Nostoc
 Oscillatoria 15
..
Usually ,composite culture containing two or more genera are
used for field inoculation since they are often superior to
single stain inoculations .

Cynobacteria produce Nitrogenase and nitrogen fixation occurs

in specialized structure called heterocyst

In which Nif-gene region becomes reoranised which is essential

for n-fixation

Heterocyst also acts as oxygen-proof compartments,which

protects Nitogenase from oxygen inactivation.

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 Azolla
• Azolla is a tiny freash water fern common in ponds , ditches and rice
fields .
• It has been used as a biofertilizer for a rice in all major rice growing
countries including India,Thailand ,Korea ,Philippines ,Brazil and
West Africa .
• Azolla is widely used in veitnam as biofertilisers but it has not
become popular in india due :-
– its sensitivity to high temperature during summers
– Necessity for assured and adequate water supply
– And susceptibility to insects and diseases
• The nitrogen fixing work is accomplished by symbiotic relationship
between the fern and BGA ,Anabena azollae.
• Azolla also provide K , P , Zn and Fe to the crop .
• It fixes 4 to 5% of nitrogen . 17
 …

• Cynobateria in addition to nitrogen fixation

,accumulates biomass ,which improves the
physical properties of soil ,produce growth
promoting substance and are useful in
reclaimation of alkaline soil.
• The are used for rice ,the inoculum being
introduced in the field about 10 days after
transplantation .
• Respositories for the culture of these micro-
organisms are , National centre for
conservation and Utilization of blue green
algae, IARI , New Delhi ,and National Facility
for Marine Cynobacteria at Bharatidasan
University , Tiruchirapalli .
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 Thiobacillus,Bacillus,etc..
• These are phosphate solubilizing
bacteria .
• They convert non-available inorganic
Phosphorus present in soil into an
available form utilized by crop plants .
• They produce iron chelating substances
e.g.,Pseudobectin called Siderophores.
• It chelate the iron present in the root
zone .
• As a result , this iron becomes non-
available to harmful micro-organisms
and , this manner ,crop plants are
protected from them.

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 C. Fungal Fertilizers

Mycorhiza
• A Mycorrhiza is a symbiotic association between a green plant and a fungus
(Glomus,.etc )
• They form two types of association with plant roots based on their location :-
 Ectomycorrhiza :- If the fungus is located at the surface of roots.
• It alleviates metal toxicity (Al and other heavy metal toxicity )at least in the case of
forest tree seedling .
 Endomycorrhiza :- If the fungus is located inside the roots.
 Fungi role:-
 These fungi converts non-available phosphorus into available form.
 They also supplies other minerals and water from soil .
 They produce growth promoting substances
 They also protect plant against soil pathogen
 Plant role :-
 It produces the organic molecules like sugar by photosynthesis and supplies to
the fungi
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  Arbuscular Mycorrhizal fungi
• An example of commercial application of Mycorrhizal fungi is found in Citrus
in U.S.A .
• Mycorrhizal fungi called AMF(Abuscular Mycorrhizal fungi).
• AMF penetrates the cortical cells of roots of a vascular plant forming
Arbuscules .
• They are inoculated with seedlings before they are transplanted in to the
field .
• The seedling with AMF survive better after transplantation (mortality rate
=zero or near zero)and grow faster .
• AMF are being exploited gainfully in India for establishing trees and other
plants on lands that are deficient in nutrients ,e.g .,wastelands or degraded
lands and on flyash overburdened sites .
• In case of crops like vegetables and flowers , It can increase 30-50% yield .
• AMF are mass produced in association with potted plants ,roots of plants
grown in aquaculture and roots cultured in vitro. 21
..

• A germplasm bank of AMF cultures was estabilished at Tata

Energy Research Institute , New Delhi in 1994.
• The bank has 240 cultures of ectomycorrhizal fungi .

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 Large scale production
• Vary critical task to choose efficient strain of nitrogen fixing
or phosphate solubilizing micro-organism .
• The main criteria for selecting an efficient nitrogen fixer are
its abilities to
I. fix a nitrogen over a range of environmental conditions
II. Compete with other strains
III. Multiply in broth and survive in carrier,
IV. Survive in seed pallets
V. Persists in soil
VI. Form nodules in case of Rhizobium
VII. Continue to fix nitrogen in presence of soil nitrogen
VIII.Migrate in soil and colonize it
IX. Be stable during storage .
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• Rhizobium inoculum is produced in shake-flasks or fermenters using appropriate
growth medium.
• After a culture attains the desired cell density , contamination is checked before
mixing it with inert and sterile carrier e,g. peat,which is subsequently used as
inoculums for coating seeds.
• Mass production of Azotobacter ,Azospirillum and p-solubilizing bacteria also incudes
starter culture preparation and and through fermenters using specific broth /media .
• BGA are multiplied in troughs or small tanks and is harvested , dried and stored in
gunny bags .algal flakes are used at 10kg/ha for rice fields .
• Azolla is vegitatively mass cultured in small field nurseries containing 10cm standing
water and irrigation channels .
• Each nursery supplied with 10g superphosphate and inoculated with 8kg fresh Azolla

to yield 40-50kg fresh Azolla per plot in 15 days .

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Seeds coated with bio-fertilizers

Mass production of AZOLLA

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.

• The longevity of bio-fertilizers depend upon storage and

distribution systems .in conditions like :-
i. Above 40 degree celsius temperature organism in biofertilizer packet
die.
ii. Damaged during transit(in hot season).etc
• BGA can be stored for longer period then others.
• BGA can survive more than 15 months while the self life of
Rhizobium,Azotobacter and Azospirillum is not more than 6
months.
• Satisfactory temperature considered is 20-25 degree celsius.
• In production units, biofertilizers are stored 4 degree celsius
temperature for longer life .
• Deep freezing is avoided ,as it damages bacterial cell membrane .

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Status
• NifTAl (U.S.A)played a major role in the production of Rhizobium
inoculants .
• The govt. of philippines implemented the National Azolla as programme
(NAAP) in 1982.For the use of Azolla as an organic fertilizers for lowland
rice fields .
• In India ,DBT , new Delhi and ICAR,New Delhi are making efforts to
popularise biofertilizers .
• Govt. has provided 20 lakh rupees for setting up of bio-fertilizers units .
• In 2005 the production of fertilizers was estimated as about 10,000 metric
tons .
• The Ministry of Agriculture ,government of India has established the
National Bio-fertilizer Development Centre at Ghaziabad with 6 regional
centres at Bangalore, Bhubaneshwar,Jabalpur,Hisar,Imphal and Nagpur.
• A range of standard quality control procedures in terms of
authentication ,maintenance , recognition of strains , test of purity etc
.,have also been prepared . 28
 Advantages
• The relevance of bio-fertilizers is increasing rapidly since chemical
fertilizers :-
I. Utilize petroleum (nitogenous fertilizers)
II. Are costly
III. Are short in supply
IV. Damage the environment .
• For example ; about 10%of ground water samples in Punjab
contain more NO3- than the max. permissible limit prescribed by
WHO.
• In contrast ,bio-fertilizers are :-
i. Low cost inputs
ii. Lead to soil enrichment
iii. Are compatible for long term sustainability.
iv. Eco-friendly
v. Pose no danger to the environment .
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 Disadvantages
• The acceptability of bio-fertilizers has been
rather low chiefly because they do not
produce ,spectacular and consistent responses
.
• The conditions under which bio-fertilizers will
be effective are not clearly established, and
quality of inoculum is an area of concern.
• The amount added by them are not enough to
adequately meet the total needs of crops for
high yields.
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Thank you

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