I want to extract various microorganisms from the rice rhizosphere, particularly bacteria and fungi. Can anybody recommend an isolation protocol specific to the said purpose?
Take 1g of rice rhizopheric soil and add 10 ml of sterilized water and mix it well. Then, prepare a serial dilution series.
Isolation of bacteria and fungi could be achieved using standard serial dilutions and spread plate methods.
Take 10 g of rice rhizospheric soil into a 300 ml of conical flask and add 90 ml of sterile distilled water to it. Shake the flask for 30 min on a rotary shaker. Add 1 ml of suspension to 10 ml vial and shake for 2 min. Spread an aliquot (100 µl) of this suspension on Nutrient agar (NA) plated petridish. Incubate the plates at 28 ± 2°C and observe the growth of the fungus periodically. Streak bacterial colonies to other NA plates and incubate at 28 ±2°C for 3 days. Pick up the well-isolated single colony and re-streak to fresh NA plates to obtain single isolation and transfer to NA slants to make pure culture.
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