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Presented by,
ROSHNI.M
PALB6078
IPhD Plant Biotechnology
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
1
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
2
Selected strains of beneficial soil microorganisms
cultured in the laboratory and packed in a suitable
carrier which increase the availability or uptake of
nutrients for plants
Biofertilizers are low cost renewable sources of plant
nutrients
Improve soil fertility and crop productivity
They are the ideal input for starting organic farming
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
3
Maintenance of soil health
Minimize environmental pollution
Cut down the use of chemical fertilizers
• Launch of ‘nitragin’- a laboratory culture of rhizobia – 1895
• In India – first study and commercial production of rhizobium
-1956 by V.N Joshi
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
4
• National biofertilizer development centre- Ghaziabhad
• Central sector scheme- NPDB- National Project on
Development and use of Biofertilizers
• Financial Assistance increased from 13 lakh to 20 lakh per unit
• Government plays a dominant role in marketing by three ways
I. State government via district level officers and village level
workers
II. State marketing federation via co-opertive bodies and
farmers
III. State agro industries co-operation via agro service centres
Wednesday,June14,2017
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Biotechnology(1+1)
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Biotechnology(1+1)
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Important Biofertilizer Producing States
6930
3257
8691
6318
2455
2925
1000
1217
Uttar Pradesh
Karnataka
Kerala
Tamil Nadu
Andhra Pradesh
Maharashtra
Madhya
Pradesh
Gujarat
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
7
Name of bio-
fertilizer
Contribution Beneficiaries
A) Nitrogen
1) Rhizobium {Sy
mbiotic}
a) Fixes 50-30 kg N/ha
b) Leaves residual nitrogen
c) Increase yield by 10 –30%
d) Maintains soil fertility
Pulses legumes: Cowpea, Green gram,
Black gram, Pea, Gram
Oil legumes: Groundnut, Soyabean
Fodderlegumes: Berseem, lucerne
Fodderlegumes:Subabul,Shisan,Wheat,Jow
ar,Bajra, Maize
2) Azotobacter a) Supplies 20-40mg N/g of carbon source
b) Promotion of growth substances like vitamins, B
Group, IAA and Gibberellic acid
c)10-15% increase in yield
d)Maintains soil fertility
e)Biological control ofplant disease, suppresses plant
pathogens
Mustard, sunflower, banana, sugarcane,
grapes,papaya,watermelon, tomato, chilly
ladyfinger,coconut,spices,flower,plantation
crops, forest sp.
3. Azospirillum a) Fixes 20-40 kg Nitrogen
b) Results in increase mineral and water uptake.
d) Vegetative growth and crop yield.
Rice, sugarcane, fingermillet, wheat,
sorghum bajra etc.;
4. Blue Green
Algae {bga}
a) 20-30 kg N/ha in submerged rice fields.
b) Production of growth substances like auxins, IAA,
giberellic acid
Rice
5. Azolla a) Fixes 40-80 kg N/ha
b) Used as green manure because of large bio-mass
Rice
Wednesday,June14,2017
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Biotechnology(1+1)
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AdvancesinMicrobial
Biotechnology(1+1)
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*Makes availability of nutrients.
*Make the root rhizosphere more lively.
*Growth Promoting Substances are produced.
*More root proliferation.
*Better germination.
*Improve quality and quantity of produce.
*Improve fertilizer use efficiency.
*More biotic and abiotic stress tolerance.
*Improve soil health.
*Residual Effect.
*Make the system more sustainable.
• Immobilization of microorganisms however
improves their shelf-life and field efficacy.
• To overcome the drawbacks of other formulations,
results in extended shelf-life, and controlled
microbial release from formulations enhancing
their application efficacy.
Wednesday,June14,2017
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Biotechnology(1+1)
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 The beads are
biodegradable and produce
no environmental pollution
 The released bacteria are
available for root
colonization immediately
at seed germination.
 Stored at ambient
temperature over a long
period without loss of
bacterial content
 storage requires a limited
space, and the quality
control of a number of
bacteria in the bead is
simple
Wednesday,June14,2017
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Biotechnology(1+1)
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Biotechnology(1+1)
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bio-fertilizers in the
market
Wednesday,June14,2017
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Biotechnology(1+1)
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Biotechnology(1+1)
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Stages of development of biofertilizers
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
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Mass production
Isolated bacterial cultures were subculture in to nutrient
broth
The cultures were grown under shaking condition at 30±2°C
The culture incubated until it reaches maximum cell
population of 10¹º to 10¹¹
Under optimum condition this population level could be
attained within 4-5 days for Rhizobium 5-7 days for
Azospirillum and 6-7 days for Azotobacter.
The culture obtained in the flask is called Starter culture
For large scale production , inoculum from starter culture is
transferred in to large flasks / fermentor and grown until
required level of cell count is reached
Rhizobium: YEMA(yeast extract mannitol Agar+
congored)
Azospirillum:Dobereiners mallic acid broth with Sodium
chloride
Azatobacter: Waksmanna No.77broth
Phosbacteria: Pikovaskys broth
Pseudomonas: Kings B broth
Trichoderma: PDB
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Biotechnology(1+1)
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Biotechnology(1+1)
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Prepare appropriate media for specific to bacterial
inoculant in required quantity
Inoculated with specific bacterial strain for aseptic condition
Incubated at 30±2ºC for 5-7 days in rotary shaker
Observe growth of the culture and estimate the population
( starter culture)
The above the media is prepared in large quantities in
fermentor
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
18
Sterilized and cooled well
Media in a fermentor is inoculated with the log phase of culture
grown in large flask (usually 1-2 % of inoculum is sufficient)
Cells are grown in fermentor by providing aeration & continuous
stirring
Broth is checked for the population of inoculated organisms
Cells are harvested with the population load of 109 cells/ml
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
19
A : Press mud ; B : Lignite : C:
Charcoal : D: Coconut Shell : E:
Rice Husk : F: Cellulose Powder :
G: Leaf Manure : H: Peat
The use of ideal carrier material is necessary for the
production of good quality of biofertilizer
Ideal carrier material should be
Cheaper in cost
Locally available
High organic matter content
No toxic chemical
Water holding capacity of more than 50%
Easy to process
Wednesday,June14,2017
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Biotechnology(1+1)
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Biotechnology(1+1)
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Preparation of inoculants packet
Neutralized and sterilized carrier material is spread in
a clean, dry, sterile metallic or plastic
Bacterial culture drawn from the fermentor is added
to the sterilized carrier and mixed well by manual or
mechanical mixer
Inoculants are packed in a polythene bags sealed
with electric sealer
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
22
Specification of the polythene bags
 Polythene bags should be of low density grade
 Thickness of bag should be around 50-75 micron
 Packet should be marked with the
 Name of the manufacture
 Name of the product
 Strain number
 The crops to which recommended
 Method of inoculation
 Date of manufacture
 Batch number
 Date of expiry
 Price
 Full address
 storage instruction
Wednesday,June14,2017
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Biotechnology(1+1)
23
23
Right type of microorganism
In Active form and in Desired numbers.
Quality of inoculants- success or failure and acceptance or
rejection
Controlled at various stages of
Production,
Marketing and
Application
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
24
Seed treatment/pelleting
Root dipping
Set treatment
Soil applications
Wednesday,June14,2017
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Biotechnology(1+1)
25
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Biotechnology(1+1)
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• Seed treatment is a most common method adopted
for all types of inoculants. The seed treatment is
effective and economic
• The seeds are treated with biofertilizer are kept in
shed for 30 mins and then seed become ready for
sowing
• Sugarcane, cut pieces of potato and base of banana
suckers
• Prepare the culture suspension by 1kg of biofertilizer
with 40-50L of water (1:50)
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
27
• The seedlings are uprooted from nursery and cleaned
their roots in water dipped in solution of biofertilizer
and kept in atleast 20 mins and transplant immediately
• Ratio about 1:10
• For root dipping : Dissolve the 1 pkt of biofertilizer with
20 litres of water (200-300 plants)
• One packet in 2 litres is sufficient to treat 200-300 sets
under cutting method
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
28
• The mixture of biofertilizer + compost + soil
applied on land before sowing of seed or
transplanting of the main field
• The mixture of biofertilizer and cattle
manure/soil sprinkled with water is then
broadcasted into the soil at the time of sowing
or at the time irrigation in standing crop
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
29
LIQUID BIOFERTILIZER
SEED TREATMENT
SOIL APPLICATION
BIOFERTIGATION
INJECTION INTO THE SOIL
Wednesday,June14,2017
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Biotechnology(1+1)
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Biotechnology(1+1)
31
Disadvantages
Biofertilizers require special care for long-term storage
because they are alive.
Must be used before their expiry date.
If other microorganisms contaminate the carrier
medium or if growers use the wrong strain, they are
not as effective.
Biofertilizers lose their effectiveness if the soil is too
hot or dry.
Wednesday,June14,2017
AdvancesinMicrobial
Biotechnology(1+1)
32

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Biofertilizers production and their applications

  • 1. Presented by, ROSHNI.M PALB6078 IPhD Plant Biotechnology Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 1
  • 2. Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 2 Selected strains of beneficial soil microorganisms cultured in the laboratory and packed in a suitable carrier which increase the availability or uptake of nutrients for plants
  • 3. Biofertilizers are low cost renewable sources of plant nutrients Improve soil fertility and crop productivity They are the ideal input for starting organic farming Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 3 Maintenance of soil health Minimize environmental pollution Cut down the use of chemical fertilizers
  • 4. • Launch of ‘nitragin’- a laboratory culture of rhizobia – 1895 • In India – first study and commercial production of rhizobium -1956 by V.N Joshi Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 4
  • 5. • National biofertilizer development centre- Ghaziabhad • Central sector scheme- NPDB- National Project on Development and use of Biofertilizers • Financial Assistance increased from 13 lakh to 20 lakh per unit • Government plays a dominant role in marketing by three ways I. State government via district level officers and village level workers II. State marketing federation via co-opertive bodies and farmers III. State agro industries co-operation via agro service centres Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 5
  • 6. Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 6 Important Biofertilizer Producing States 6930 3257 8691 6318 2455 2925 1000 1217 Uttar Pradesh Karnataka Kerala Tamil Nadu Andhra Pradesh Maharashtra Madhya Pradesh Gujarat
  • 8. Name of bio- fertilizer Contribution Beneficiaries A) Nitrogen 1) Rhizobium {Sy mbiotic} a) Fixes 50-30 kg N/ha b) Leaves residual nitrogen c) Increase yield by 10 –30% d) Maintains soil fertility Pulses legumes: Cowpea, Green gram, Black gram, Pea, Gram Oil legumes: Groundnut, Soyabean Fodderlegumes: Berseem, lucerne Fodderlegumes:Subabul,Shisan,Wheat,Jow ar,Bajra, Maize 2) Azotobacter a) Supplies 20-40mg N/g of carbon source b) Promotion of growth substances like vitamins, B Group, IAA and Gibberellic acid c)10-15% increase in yield d)Maintains soil fertility e)Biological control ofplant disease, suppresses plant pathogens Mustard, sunflower, banana, sugarcane, grapes,papaya,watermelon, tomato, chilly ladyfinger,coconut,spices,flower,plantation crops, forest sp. 3. Azospirillum a) Fixes 20-40 kg Nitrogen b) Results in increase mineral and water uptake. d) Vegetative growth and crop yield. Rice, sugarcane, fingermillet, wheat, sorghum bajra etc.; 4. Blue Green Algae {bga} a) 20-30 kg N/ha in submerged rice fields. b) Production of growth substances like auxins, IAA, giberellic acid Rice 5. Azolla a) Fixes 40-80 kg N/ha b) Used as green manure because of large bio-mass Rice Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 8
  • 9. Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 9 *Makes availability of nutrients. *Make the root rhizosphere more lively. *Growth Promoting Substances are produced. *More root proliferation. *Better germination. *Improve quality and quantity of produce. *Improve fertilizer use efficiency. *More biotic and abiotic stress tolerance. *Improve soil health. *Residual Effect. *Make the system more sustainable.
  • 10. • Immobilization of microorganisms however improves their shelf-life and field efficacy. • To overcome the drawbacks of other formulations, results in extended shelf-life, and controlled microbial release from formulations enhancing their application efficacy. Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 10
  • 11.  The beads are biodegradable and produce no environmental pollution  The released bacteria are available for root colonization immediately at seed germination.  Stored at ambient temperature over a long period without loss of bacterial content  storage requires a limited space, and the quality control of a number of bacteria in the bead is simple Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 11
  • 15. Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 15 Mass production Isolated bacterial cultures were subculture in to nutrient broth The cultures were grown under shaking condition at 30±2°C The culture incubated until it reaches maximum cell population of 10¹º to 10¹¹ Under optimum condition this population level could be attained within 4-5 days for Rhizobium 5-7 days for Azospirillum and 6-7 days for Azotobacter. The culture obtained in the flask is called Starter culture For large scale production , inoculum from starter culture is transferred in to large flasks / fermentor and grown until required level of cell count is reached
  • 16. Rhizobium: YEMA(yeast extract mannitol Agar+ congored) Azospirillum:Dobereiners mallic acid broth with Sodium chloride Azatobacter: Waksmanna No.77broth Phosbacteria: Pikovaskys broth Pseudomonas: Kings B broth Trichoderma: PDB Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 16
  • 17. Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 17 Prepare appropriate media for specific to bacterial inoculant in required quantity Inoculated with specific bacterial strain for aseptic condition Incubated at 30±2ºC for 5-7 days in rotary shaker Observe growth of the culture and estimate the population ( starter culture) The above the media is prepared in large quantities in fermentor
  • 18. Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 18 Sterilized and cooled well Media in a fermentor is inoculated with the log phase of culture grown in large flask (usually 1-2 % of inoculum is sufficient) Cells are grown in fermentor by providing aeration & continuous stirring Broth is checked for the population of inoculated organisms Cells are harvested with the population load of 109 cells/ml
  • 19. Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 19 A : Press mud ; B : Lignite : C: Charcoal : D: Coconut Shell : E: Rice Husk : F: Cellulose Powder : G: Leaf Manure : H: Peat The use of ideal carrier material is necessary for the production of good quality of biofertilizer Ideal carrier material should be Cheaper in cost Locally available High organic matter content No toxic chemical Water holding capacity of more than 50% Easy to process
  • 21. Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 21 Preparation of inoculants packet Neutralized and sterilized carrier material is spread in a clean, dry, sterile metallic or plastic Bacterial culture drawn from the fermentor is added to the sterilized carrier and mixed well by manual or mechanical mixer Inoculants are packed in a polythene bags sealed with electric sealer
  • 22. Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 22 Specification of the polythene bags  Polythene bags should be of low density grade  Thickness of bag should be around 50-75 micron  Packet should be marked with the  Name of the manufacture  Name of the product  Strain number  The crops to which recommended  Method of inoculation  Date of manufacture  Batch number  Date of expiry  Price  Full address  storage instruction
  • 23. Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 23 23 Right type of microorganism In Active form and in Desired numbers. Quality of inoculants- success or failure and acceptance or rejection Controlled at various stages of Production, Marketing and Application
  • 25. Seed treatment/pelleting Root dipping Set treatment Soil applications Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 25
  • 26. Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 26 • Seed treatment is a most common method adopted for all types of inoculants. The seed treatment is effective and economic • The seeds are treated with biofertilizer are kept in shed for 30 mins and then seed become ready for sowing
  • 27. • Sugarcane, cut pieces of potato and base of banana suckers • Prepare the culture suspension by 1kg of biofertilizer with 40-50L of water (1:50) Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 27
  • 28. • The seedlings are uprooted from nursery and cleaned their roots in water dipped in solution of biofertilizer and kept in atleast 20 mins and transplant immediately • Ratio about 1:10 • For root dipping : Dissolve the 1 pkt of biofertilizer with 20 litres of water (200-300 plants) • One packet in 2 litres is sufficient to treat 200-300 sets under cutting method Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 28
  • 29. • The mixture of biofertilizer + compost + soil applied on land before sowing of seed or transplanting of the main field • The mixture of biofertilizer and cattle manure/soil sprinkled with water is then broadcasted into the soil at the time of sowing or at the time irrigation in standing crop Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 29
  • 30. LIQUID BIOFERTILIZER SEED TREATMENT SOIL APPLICATION BIOFERTIGATION INJECTION INTO THE SOIL Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 30
  • 31. Wednesday,June14,2017 AdvancesinMicrobial Biotechnology(1+1) 31 Disadvantages Biofertilizers require special care for long-term storage because they are alive. Must be used before their expiry date. If other microorganisms contaminate the carrier medium or if growers use the wrong strain, they are not as effective. Biofertilizers lose their effectiveness if the soil is too hot or dry.