Presented By, Roshni.M Palb6078 Iphd Plant Biotechnology

IPhD Plant Biotechnology
ROSHNI.M
Presented by,
PALB6078
Advances in Microbial
Wednesday, June 14, 2017
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Biotechnology (1+1)
Selected strains of beneficial soil microorganisms
cultured in the laboratory and packed in a suitable
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carrier which increase the availability or uptake of
nutrients for plants
2
Biofertilizers are low cost renewable sources of plant
nutrients

Improve soil fertility and crop productivity
They are the ideal input for starting organic farming
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Maintenance of soil health
Minimize environmental pollution
Cut down the use of chemical fertilizers
3
• Launch of ‘nitragin’- a laboratory culture of rhizobia – 1895
• In India – first study and commercial production of rhizobium
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-1956 by V.N Joshi
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
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• 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
5
Important Biofertilizer Producing States

Andhra Pradesh
1000
Uttar Pradesh
1217
Maharashtra Karnataka
2925 6930
Madhya
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Pradesh 2455
3257
Kerala
6318
Gujarat
8691
Tamil Nadu
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Name of bio- Contribution Beneficiaries
fertilizer
A) Nitrogen a) Fixes 50-30 kg N/ha Pulses legumes: Cowpea, Green gram,
1) Rhizobium {Sy b) Leaves residual nitrogen Black gram, Pea, Gram
mbiotic} c) Increase yield by 10 –30% Oil legumes: Groundnut, Soyabean

d) Maintains soil fertility Fodderlegumes: Berseem, lucerne
Fodderlegumes:Subabul,Shisan,Wheat,Jow
ar,Bajra, Maize
2) Azotobacter a) Supplies 20-40mg N/g of carbon source Mustard, sunflower, banana, sugarcane,
b) Promotion of growth substances like vitamins, B grapes,papaya,watermelon, tomato, chilly
Group, IAA and Gibberellic acid ladyfinger,coconut,spices,flower,plantation
c)10-15% increase in yield crops, forest sp.
d)Maintains soil fertility
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e)Biological control ofplant disease, suppresses plant
pathogens
3. Azospirillum a) Fixes 20-40 kg Nitrogen Rice, sugarcane, fingermillet, wheat,

b) Results in increase mineral and water uptake. sorghum bajra etc.;
d) Vegetative growth and crop yield.
4. Blue Green a) 20-30 kg N/ha in submerged rice fields. Rice

Algae {bga} b) Production of growth substances like auxins, IAA,
giberellic acid 8
5. Azolla a) Fixes 40-80 kg N/ha Rice
b) Used as green manure because of large bio-mass
*Makes availability of nutrients.

*Make the root rhizosphere more lively.
*Growth Promoting Substances are produced.
*More root proliferation.
*Better germination.
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*Improve quality and quantity of produce.
*Improve fertilizer use efficiency.
*More biotic and abiotic stress tolerance.
*Improve soil health.
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*Residual Effect.
*Make the system more sustainable.
• Immobilization of microorganisms however
improves their shelf-life and field efficacy.
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• To overcome the drawbacks of other formulations,
results in extended shelf-life, and controlled
microbial release from formulations enhancing
their application efficacy.
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 The beads are
biodegradable and produce
no environmental pollution
 The released bacteria are
available for root
colonization immediately
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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 11
market
bio-fertilizers in the
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Biotechnology (1+1)
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Biotechnology (1+1)
Stages of development of biofertilizers
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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
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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 15
required level of cell count is reached
Rhizobium: YEMA(yeast extract mannitol Agar+
congored)
Azospirillum:Dobereiners mallic acid broth with Sodium
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chloride
Azatobacter: Waksmanna No.77broth
Phosbacteria: Pikovaskys broth
Pseudomonas: Kings B broth

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Trichoderma: PDB
Prepare appropriate media for specific to bacterial

inoculant in required quantity
Inoculated with specific bacterial strain for aseptic condition
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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
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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)
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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

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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
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Water holding capacity of more than 50%
Easy to process
A : Press mud ; B : Lignite : C:

Charcoal : D: Coconut Shell : E:
Rice Husk : F: Cellulose Powder :
G: Leaf Manure : H: Peat
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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
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to the sterilized carrier and mixed well by manual or
mechanical mixer
Inoculants are packed in a polythene bags sealed
with electric sealer
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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
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 Strain number
 The crops to which recommended
 Method of inoculation
 Date of manufacture
 Batch number
 Date of expiry
 Price
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 Full address
 storage instruction
 Right type of microorganism
 In Active form and in Desired numbers.
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Quality of inoculants- success or failure and acceptance or
rejection
Controlled at various stages of

Production,
Marketing and
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Application
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Seed treatment/pelleting
Root dipping
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Set treatment
Soil applications
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• Seed treatment is a most common method adopted
for all types of inoculants. The seed treatment is
effective and economic
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• The seeds are treated with biofertilizer are kept in
shed for 30 mins and then seed become ready for
sowing
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• Sugarcane, cut pieces of potato and base of banana
suckers
• Prepare the culture suspension by 1kg of biofertilizer
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with 40-50L of water (1:50)
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• 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
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• 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 28
• The mixture of biofertilizer + compost + soil
applied on land before sowing of seed or
transplanting of the main field
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• The mixture of biofertilizer and cattle
manure/soil sprinkled with water is then
broadcasted into the soil at the time of sowing
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or at the time irrigation in standing crop
SEED TREATMENT
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SOIL APPLICATION
LIQUID BIOFERTILIZER
BIOFERTIGATION
INJECTION INTO THE SOIL 30

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
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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.
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Biotechnology (1+1)

Presented By, Roshni.M Palb6078 Iphd Plant Biotechnology

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IPhD Plant Biotechnology

Wednesday, June 14, 2017

They are the ideal input for starting organic farming

Minimize environmental pollution

Cut down the use of chemical fertilizers

Wednesday, June 14, 2017

Wednesday, June 14, 2017

3. Azospirillum a) Fixes 20-40 kg Nitrogen Rice, sugarcane, fingermillet, wheat,

4. Blue Green a) 20-30 kg N/ha in submerged rice fields. Rice

Wednesday, June 14, 2017

Wednesday, June 14, 2017

Azospirillum:Dobereiners mallic acid broth with Sodium

Azatobacter: Waksmanna No.77broth

Phosbacteria: Pikovaskys broth

Pseudomonas: Kings B broth

Wednesday, June 14, 2017

Inoculated with specific bacterial strain for aseptic condition

Observe growth of the culture and estimate the population

Wednesday, June 14, 2017

Broth is checked for the population of inoculated organisms

Cells are harvested with the population load of 109 cells/ml

Wednesday, June 14, 2017

A : Press mud ; B : Lignite : C:

Wednesday, June 14, 2017

Wednesday, June 14, 2017

 In Active form and in Desired numbers.

Controlled at various stages of

• Prepare the culture suspension by 1kg of biofertilizer

INJECTION INTO THE SOIL 30

Wednesday, June 14, 2017

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