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Recent trends in Microbial Fertilizers (Microbial Inoculant ). Dr.A.M.Deshmukh, Prof and Head, Dept of Microbiology, Dr.Babasaheb Ambedkar Marathwada University,Subcentre Osmanabad.M.S.s. Biofertilizer. Chemical fertilizer –Problem
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Recent trends in Microbial Fertilizers (Microbial Inoculant) Dr.A.M.Deshmukh, Prof and Head, Dept of Microbiology, Dr.Babasaheb Ambedkar Marathwada University,Subcentre Osmanabad.M.S.s
Biofertilizer • Chemical fertilizer –Problem • Petroleum crisis • Wastage in soil, • Foreign currency, • Pollution, • Health hazards Decrease in soil fertility • More demand • Costly
Advantages • Cheap • Reduce the demand • No pollution problem • Production –simple • Also supply plant growth hormone, • Supply other nutrients • Control plant pathogens • Prevent erosion of soil, • Increase availability of chemical fertilizer
Types • Nitrogen fixing- 1.Symbiotic Rhizobium,Frankia, 2.Non symbiotic Azotobacter,Blue green algae,Azospirillum,Azolla, Acetobacter,Herbaspirillum
Types • Phosphate supplier 1.Phosphate absorber 2.Phosphate solubilizer • Sulfur supplier • Potash solubilizers • Organic matter decomposer • Microbial cell mass • Liquid biofertilizer
Rhizobium • Gram negative motile bacteria • Root nodules in leguminous plants • Leg haemoglobin Red colored • Effective nodules are large pink and firm • Bacteroids in crushed root nodules • Grow on congored mannitol agar • Fix atmospheric nitrogen • Bradyrhizobium slow grower • Nitrogenase enzyme-nif gene
Rhizobium-production • Isolation,Confirmation of nodulation ability,Selection of suitable strains,Preparation of seed culture,inoculants preparation,carrier preparation and mixing with carrier,Stabilization,packaging,labeling and storage-Marketing
Selection of strain • Bacteriophage resistant • Good shelf life • Good nitrogen fixation ability • Pesticide resistant • Competitive • Withstand extreme sdoil and weather condition • Better host infection ability
Equipment • Autoclave • Hot air oven • Rotary shaker or fermentor • Refrigerator, • Mechanical mixer, Sealer, Steel trays • Balance • Chemicals and glasswares
One tonne production • 4 liter seed culture-106-107 cells/ml • 396 liter yeast extract mannitol broth-sterilized • 590 carrier material -sterilized • 10 kg gum solution-sterilized • 5000 polypropylene packets 50-75 microns with 200 g capacity
Carrier • peat.>Lignite>Charcoal>Farm yard manure, • Rice husk,press mud • Lime addition 10-15% • Autoclaved 4 h on first day 1 hr on subsequent two days at 1210C • Gamma irradiation
Adhesive • Gum Arabic,Honey,CMC,Powder milk,Sugar,Jaggery
Space • Five rooms 10x15 feet, cement floor 1.Microbial culture 2.Production 3.Carrier 4.Mixing and stabilization 5.Packeged product
Frankia-Actinorhiza • About 200 plant species Alnus,Casurina,Myrica,Discaria-Angiosperm-Forest crop • Nodulated nonlegumes • Gram positive, filamentous Slow grower form visible colony within 3-6 months • Nodules –Multilobed amber or whitish,woody 5-10 mm in diameter-about 1 g in weight
Azotobacter • Heterotrophic free living nitrogen fixing bacteria gram negative bacteria • Produce resistant structure Cyst • Produce growth promoting substances • A.chrococcum,A.vinelandii,A.beijerinckia A. Insignis,A. macrocytogenes, • A.nitrocaptans • Many strains are fungi static • Isolation-Nitrogen free mannitol medium
Azolla • A.Carolina,A.filiculodes,A.mexicana, A.microphylla,A.nilotica,A. pinnata • Anabaena azollae • 94% water,1%minerals,5%Nitrogen • Decompose in soil within 7-8 days • Stop growing when touches to soil • Difficult for transportation
Azolla • Microplot 3x1x0.2m lined with plastic paper Soil 10-15kg Cow dung 5kg Water 10mm in height Phosphates 40g Azolla 500g Temperature Pesticide • Azolla mat
BGA • 4% are nitrogen fixer • Heterocyst • 30-70% are blue green algae in moist soil • Mainly applied in rice crop
BGA • Small pit 1x1meter Lined with polyethylene sheet 10 Kg sieve soil, 250 g Super phosphate, Water10cm in height Inoculum Pesticide Na molybdate 1 g Lime Incubation 3 weeks • Algal mass
Azosprillum • Dr.Doberiner Brazil 1975 • A.brasilense,A.lipoferum,A.halopraeferans,A.amazonens • Plant-Many grasses,maize,wheat,sorghum,rice, mustard • Growth at 320-350C
Azospirillum • Isolation-0.5 cm root piece-Surface sterilized-Washed repeatedly in sterile saline water • Placed on semi solid Nitrogen free malate medium • Incubate- White pellicle
Herbaspirillum • Associative symbiont-Sugar cane • Uptake of nitrates potassium,phosphates
Acetobacter-Gluconobacter • A.diazotrophicus • Roots,stems and leaves of sugarcane • Antaginists red-rot disease • Tolerate high sugar concentration and low pH
Mycorrhizae • Uptake of nutrients Zinc, Iron, manganese, sulphur,phosphorus Production-Seeds in tray (sterile) in sterile soil and sand+ inoculum Temp-25-280C,Humidity 70-80%Light intensity 10-15 k Lux - Watered regularly After 60 day-crush roots-use as inoculum
Phosphate Solubilizers • Bacteria • Bacillus polymyxa,B.subtilis,B.subtilis, B.megaterium,Pseudomonas, Achromobacter • Fungi • Aspergillus avmori,A. niger, A. flavus,Penicillium digitatum • Actinomycetes Streptomyces,Actinomyces
Mechanism of PSM • Production of organic acids Lactic acid,citric acid butyric acid succunic acid,fumaric acid • CO2 liberated reacts with water and produce acid • Phosphatase enzyme release organic phosphate • H2S react with insoluble phosphate and produce ferrous sulphide
Potash solubilizing bacteria • Frateuria aurantia • 2.5 kg/ha mix with 200-500 kg FYM
Sulfur oxidisers • Thiobacillus ferrooxidans, • T.novellus,Ferrobacillus ferroxidans • Convert insoluble sulphides to soluble sulphate
Microbial cell mass • Many industries use microbial culture • Waste after recovery is microbial cell mass • Penicillium from penicillin factory • Contain chitin in cell wall • Increase organic matter of soil • Chitin chelates metals like Fe, cu & zn • Contain various enzymes
Application • Treatment before sowing • Seedling treatment before transplantation • Treating the soil
Application –Seed treatment • 50g crude sugar/200g gum arabic+500ml water • Heat for 15 min • Cooled • 200g biofertilizer • 10Kg seed • Dried in shade for 10-15 min • Mung bean,cowpea,Pigeon pea,lentil-20g/kg • Chick pea 30-40g/kg
Deeping seedling • 1-2 kg in 15 lit of water • 250g in 1 lit of cow dung slurry-VAM • Roots of seedling from 10-15 kg seed-for10-30min • Sugarcane sets • 1-2 kg of biofertilizer in 50-100 liter of water
Soil treatment • 500-800g biofertilizer+ 10-15 kg of FYM OR Compost-24 hr before earlier
Application • BGA- • Dried flakes-10kg/ha-10days after transplanting rice in 2-3 cm standing water • Azolla • 3-4 tonnes /ha before transplanting • 2-5 kg/ha in the crop • VAM • 3-5cm below the seed • Seedbed 5 kg/25 sq. m.
Liquid Biofertilizers • Shelf life more • Can have higher count, • No contamination • Easy for application • Easy to handle • Attractive packaging • Good field response • Easy sterilization
Labeling • Name of biofertilizer • Manufacturing date • Names of recommended crops • Batch no. • Expiry date • Weight, • Precautions for storage • Name and address of manufacturer
Quality control • Ph-6.5-7.5 • Passed through 85 mesh sieve • Moisture 30-40% • Number- as per standard • 3 pocket/1000
Why failure? • Environmental factors-pH,Organic matter,Temperature,Moisture content • Pesticide application • Application after expiry • Stored at high temp • Competition-Resident flora • Non specific • Poor quality
Precaution • Store in shade • Store at cool place • Apply pesticides and then biofertilizer • Apply in moist soil • Add molybdenum for nitrogen fixer • Prevent contamination • Prevent holes in bags • Use specific biofertilizer
Constrains • Untrained labor, • Poor standards • No rules and regulations • Uneducated farmers • 20% loss due to expiry • No storage facility • No good carrier • No marked effect