2.1. deffered spot assay were performed for yeast strains.

2.1.
Revival of Bacterial Cultures

A total 79 strains of indigenously isolated Bacillus sp.
were revived from old glycerol stocks.A loopful of glycerol stock was streaked
on pre-incubated nutrient agar plates and incubated at 370C for 24
hours. Next day the isolated colony was transferred to nutrient agar slant and
kept at 40C in refrigerator for short term storage. Whereas for long
term storage 15 % v/v glycerol stocks were prepared by mixing 24hour old
bacterial culture in nutrient broth with 30% sterile glycerol in a ratio 1:1
and stored at -20o C.

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M2.2
ReIdentification of Bacterial Isolates

M2.3
Revival and Maintenance of Fungal Cultures

All fungal cultures were revived from time to time  and maintained on Sabroud Dextrose agar and
purity was checked through Lactophenol Cotton blue mount. All fungal stocks
were stored on SDA slants at 40C for short term storage. Soil stocks
were also prepared for long term storage of fungal culture. Sterile soil was
inncoulated with the fresh fungal spore suspension in sterile saline( approx.
109 /ml) and incubated at room temperature.

M2.4
PrimaryScreening for antifungal activity

To check the ability of producer for antifungal activity,
both simultaneous and deffered spot assay were performed for yeast strains.

M2.4.1Simaltaneous
Spot Assay for Yeast Sp.

Producer bacterial isolates were spot inoculated on preincubated
nutrient agar plate with sterile inoculating needle. The plate is then
overlayed with soft SDA (0.7% agar) seeded with yeast spore suspension containing
approx. 109 spores/ml and allowed to solidify and incubated at room
temperature for 48 hours.

M2.4.2
Deffered Spot Assay for Yeast Sp.

Indicator bacterial cultures were spotted on pre-incubated
nutrient agart plates and incubated at 370C for 24 hours. The
following day, the same plate is overlaid with soft SDA containing yeast spore
suspension (109 spores/ml) ,allowed to solidify and incubated at
room temperature for 48 hours .

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