OM SAKTHI

ADHIPARASAKTHI HORTICULTURAL COLLEGE

G.B. NAGAR, KALAVAI, RANIPET DISTRICT,
TAMILNADU – 632506.
AGM-451 BIO-INOCULANTS PRODUCTION
TECHNOLOGY(0+10)

COURSE TEACHER : Mrs.M.CHITRA

S.No. ID No. Students name
1. 2019022001 AAKASH.K
2. 2019022004 AARTHI.S
3. 2019022006 ABISHEK.S
4. 2019022012 ARUN KUMAR.E
5. 2019022014 ASHWINI.M
6. 2019022017 BOOMIKA.V
7. 2019022022 DHANAVIJAYAKUMAR.D
8. 2019022025 DHAYALAN.E
9. 2019022028 DIVYASREE.N.R
10. 2019022031 GOKUL.R
S.No. ID No. Students name
11. 2019022033 GUNNAL.S.A
12 2019022035 GURUPRASATH.R
13 2019022039 HARISH KUMAR.A
14 2019022040 HELAL.H
15 2019022041 HEMAVATHY.D
16 2019022042 ISHWARYALAKSHMI.G
17 2019022044 JAYAGANESH.A
18 2019022045 JEEVITHA.V
19 2019022047 KAMALESHRAJAN.H
20 2019022061 MAMTHA.V
S.No. ID No. Students name
21 2019022064 MOHANRAJ.D
22 2019022074 OVIYA.M.J
23 2019022086 PRIYANKA.M
24 2019022098 SHIVA HARSHITH.T
25 2019022099 SIVANESH.B
26 2019022107 TAMILARASI.S
27 2019022111 VIGNESHVARA.R
28 2019022112 VIKAASH.K
29 2019022117 PAVATHARANI.M
30 2019022119 SUCHITHRA.R
 INTRODUCTION
Bioinoculants:
Bioinoculants are the beneficial soil
amendments that use microbes for promoting
plant growth and development.
BIOFERTILIZER DEFINITION(VESSEY, 2003):
 Biofertilizer can be defined as biological products
containing living microorganisms
 Applied to seed, plant surfaces, or soil
 Promote growth by several mechanisms such as
increasing the supply of nutrients, increasing root
biomass or root area and increasing nutrient uptake
Sample Footer Text
capacity of the plant 5
megatheria VA Mycorrhizae
striata
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 IMPORTANCE OF BIOFERTILIZER:

Biofertilizer production aims to supplement the soil with the essential nutrients
that already exists in it

Maintain microbial consortia in soil

Maintenance of symbiotic relationships

Increase yield by 10 to 25%

Promotes crop health to certain extent

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 Rhizobium
(BEIJERNICK,1888)

Domain Bacteria  Endosymbiotic nitrogen fixers

Phylum Pseudomonadota
 Gram negative rod shaped soil
Class Alphaproteobacteria bacteria
Order Hyphomicrobiales
 Species:
Family Rhizobiaceae
Genus Rhizobium R.leguminosarum,R.alamii,
R.japonicum,R.phaseoli,
R.trifoli and R.
metallidurans

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 ISOLATION OF Rhizobium
• PREPARATION OF ROOT SUSPENSION(BEIJERNICK,1888):

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• SERIAL DILUTION (ROBERT KOCH,1883):

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• PLATING TECHNIQUE: STREAK PLATE

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 Incubation Colony
Media:
characters :
Microscopic
CRYEMA period: 3 – 5 view
days gummy raised
(Vincent 1974)
colonies

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PURIFICATION OF Rhizobium

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 Azospirillium
(DR. JOHANNA
DÖBEREINER,1970)
Domain Bacteria • Spiral-shaped associative N-fixing
bacteria
Phylum Pseudomonadota
• Distributed in soils and grass roots
Class Alphaproteobacteria
Order Rhodospirillales • Gram negative bacteria

Family Azospirillaceae • Species : Azospirillum brasilense and

Azospirillum lipoferum
Genus Azospirillum
• Produces hormones such as indole
acetic acid (IAA), gibberellic acid
(GA), cytokinins and vitamins

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 ISOLATION OF Azospirillum

• STERILIZATION OF PADDY ROOT BITS:

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• INOCULATION OF ROOT BIT:

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Media:Malic acid Colony characters:
or NFB semi-solid Media colour
Incubation period:
media(Dobereiner changes from
2 – 3 days @ 26 to
and Day, 1976) yellow to blue and
32c
formation of white
pellicle

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 Azotobacter
(BEIJERNICK,1901)
Domain Bacteria
Phylum Pseudomonadota
• Non-symbiotic, aerobic, free-living, n-
Class Gammaproteobacteria
fixing soil bacterium
Order pseudomonadales • Rod shaped Gram negative bacteria
Family Pseudomonadaceae • Species are : Azotobacter insignis,
Genus Azotobacter A.Beijerinckii, A.Chroococcum,
A.macrocytogens, A.Paspali And
A.Vinelandi

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 ISOLATION OF Azotobacter

• ROOT ISOLATION OF AZOTOBACTER:

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 Incubation Black to brown
Media: Microscopic
period: 3 – 4 coloured
Waksmann View
days @ 26 to pigmented
NO.77 media
32c colonies

Sample Footer Text 20

 CASE STUDY OF GROWTH OF Azotobacter IN
DIFFERENT METHODS

Isolation through serial Isolation through root isolation Isolation through root isolation
dilution at 10−3 with sterilization techniques with cleaning the root bits in
distilled water

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 Glucanoacetobacter
(Cavalcante and
Dobereiner 1988)

Domain Bacteria • Rod-shaped,gram negative, aerobic,

Phylum Pseudomonadota N-fixing bacteria
Class Alphaproteobacteria • Capable of growth at Ph 3
Order Rhodospirillales • Inoculation with acetobacter is
Family Acetobacteraceae recommended for sugar cane

Genus Glucanoacetobacter • It is also called as black urea

• Also solubilize insoluble forms of P.

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 ISOLATION OF Glucanoacetobacter
(Cavalcante and Dobereiner, 1988)
• PREPARATION OF ROOT SUSPENSION:

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TRANSFER OF ROOT SUSPENSION TO TEST TUBE WITH LGI MEDIA

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 Colony
Media:LGI semi- Incubation period: characters:orange
solid media 2 – 3 days @ 26 to coloured pellicle is
32 formed on the test
tube

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 CASE STUDY ON GROWTH OF
Glucanoactobacter IN DIFFERENT MEDIA
COMPOSITION

WITHOUT CANE JUICE WITH CANE JUICE

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 PHOSPHOBACTERIA

SPECIES:
• Bacillus megatherium var. phosphaticum,
• Bacillus polymyxa
• Bacillus subtilis
• Pseudomonas striata

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 ISOLATION OF PHOSPHOBACTERIA FROM
SOIL
Serial dilution (Robert Koch,1883):

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PLATING TECHNIQUE:

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 Incubation
Media: Colony
period: 2 – 3
pikovskaya characters:clear
days @ 26 to
media halo zones
32c

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 Plant Growth Promoting Rhizobacteria(KLOEPPER
AND SCHROTH,1970)
PGPR EXAMPLE :
• Azotobacter ,
• Azospirillum,
• Azoarcus,
• Klebsiella,
• Arthrobacter,
• Pseudomonas fluorescence,
• Rhizobium,
• Bacillus.
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 ISOLATION OF PGPR
SERIAL DILUTION (ROBERT KOCH,1883) AND PLATING
TECHNIQUE:

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Media:kings B Incubation Colony character Microscopic
medium period: 3 – 4 :flurosence view
days @ 26 to 32c colonies

Sample Footer Text 33

 Pink Pigmented Facultative
Methylotrophs(PPFM)
( GREEN AND BOUSEFEILD,1983)
Domain Bacteria  Aerobic, gram-negative
Phylum Pseudomonadota bacteria
Class Alphaproteobacteria
 Rod shaped bacteria
Order Hyphomicrobiales
Family Methylobacteriaceae PPFM EXAMPLES :
Genus Methylobacterium Methylobacterium, Methylirubrum.

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LEAF IMPRINT TECHNIQUE
Media:Ammonium
Colony
Mineral Salt Incubation period:5
characters:Pink
Media(Green and days @ 26 to 32c
coloured colonies
Bousefeild,1983)

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MASS PRODUCTION OF BACTERIAL
BIOFERTILIZERS

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 MASS PRODUCTION OF BACTERIAL INOCULANTS

Broth culture Mixing of broth

culture with carrier
material
Lignite+calcium Packaging
carbonate
 EXAMINATION OF AM INFECTION IN
ROOTS

INTRODUCTION:
• Mycorrhiza –means root fungus, reported by A.B.Frank in 1885.

• This fungus absorbs the available p from soil and transports

directly to root cortex.
CLASSIFICATION OF MYCORRHIZA FUNGI

Mycorrhiza

Endomycorrhiza Ectomycorrhiza

Arbuscules
Vesicles

Ericiod Orchid Arbuscular

 STEPS IN EXAMINATION OF AM FUNGI
Root Clearing and Staining Technique

Bleaching

Acidification

Staining

AM Fungi in
Destaining Microscopic
View
Microscopic Observation
 RECOVERY OF VAM FUNGAL SPORES
FROM SOIL
TECHNIQUE: WET SIEVING AND DECANTING( GERDEMANN
AND NICOLSON 1963)
 MASS PRODUCTION OF AM FUNGI
(MOSSE AND HEPPER,1975)

Material required:
• 5 kg - Sterile Soil
• 25 kg - Vermiculite
• 1kg -VAM Inoculum
• Maize seeds
• Nutrients:
• 2g of Urea
• 2g of Single Super
Phosphate
• 1g of MOP
 AZOLLA
Kingdom Plantae
• Azolla spp. are free floating Clade Tracheophyte
freshwater ferns.
Division Polypodiophyta
• Float on the water surface
individually or in mats. Class Polypodiopsida
• Also known as duckweed ferns. Order Salviniales
• Sizes (dia)- ranges from 1/3 to 1 inch
(1-2.5 cm ) to 6 inches (15 cm or Family Salviniaceae
more.
Genus Azolla
• SPECIES OF AZOLLA FRONDS
• A.filiculoides
• A.caroliniana
• A.mexicana
• A.microphylla (U.P)
• A.nilotica
• A.pinnata (TN,AP)
• Anabaena azollae
• A.spiculata
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DIFFERENT SPECIES OF AZOLLA

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MASS PRODUCTION OF AZOLLA IN CEMENT
TANKS
Materials required
Cement tank /pits

cattle dung

super phosphate
• fresh azolla inoculum(Azolla pinnata, Azolla spiculata is used as inoculum – source of
inoculum – KVK virinjipuram)
 CHARACTERISTICS OF Azolla spiculata

• Azolla spiculata(mosquito fern) forms dense mats on the surface of still or slow-moving water
bodies
• Under the microscope, the leaves are intricately arranged in a compact and overlapping pattern,
forming a floating mat on the water surface
• Each individual leaflet is about 0.5 to 1.5 millimeters in size and has a flattened shape
• The upper surface of the leaflets is covered with a waxy cuticle, giving them a glossy appearance
• The lower surface of the leaflets contains numerous tiny root-like structures called rhizoids,
which aid in absorbing nutrients from the water

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MASS PRODUCTION OF AZOLLA
 ESTIMATED BCR RATIO
Fixed cost (non-recurring)
Equipment Quantity required Cost (Rs)
Glass double distillation set 5L/hr 1 11,000
capacity
Incubator 1 6999
Compound microscope 1 10000
Refrigerator 1 24990
Inoculation chamber with UV 1 35000
Polythene sealer 1 840
Chemical balance 1 3460
Total 92289

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Variable cost (recurring cost)
Material Quantity Cost
Carrier material 25 kg 1250
Broth 20 L 310
Polythene bags 1000 1225
Recurring expenses - 1500
Total 4282.5

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Salary component:
Salary components Value(Rs)
One microbiologist (skilled) per month 15,000
One production assistant (unskilled) per 8,000
month
Miscellaneous 1250
Total 17,050

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• Production Cost for 500 Packets = Rs.30839
• Gross returns for 500 packets = Rs.60000
• Thus, Net Return is Rs.29161

BCR = 1:2.1

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 THANK
YOU

Sample Footer Text 54