Rhizobium are soil bacteria that fix
nitrogen after becoming established inside root nodules of legumes. Rhizobia require a plant host. Cannot independently fix nitrogen.they break down complex carbon compounds for energy.most living creatures depends upon inorganic froms of nitrogen, rhizobia possess a very special enzyme called nitrogenase. This enzyme allows them to convert N2 gas into ammonium.
Rhizobia , establishes a symbiotic relationship with a legume plant, providing nitrogen in exchange for a protected environment.
Material and Method
Instrument:
1. Bunsen burner
2. Laminar air flow
3. Microscope
4. Oven
5. Incubators
6. Refrigerators
7. Autoclave
8. pH indicator
9. Weighing machine
TOOL
Inoculating loop
GLASSWARE
1. Petridishes
2. Conical flask
3. Beakers
4. Measuring cylinder
5. Pipettes
6. Test tubes
Preparation of culture media
The growth of an organism on a midium is called culture. The food base that support the growth of an organism is called culture medium
Composition of YEMA medium for Rhizobium
Ingredients Gram/Lt. Mannitol 10.0 gm
Dipotassium phosphate 0.5 gm. Magnesium sulphate. 0.2gm. Sodium chloride. 0.1 gm. Yeast extract 1.0gm
Agar. 20.0gm
Congo red 0.025gm
Distilled water. 1000ml
PH of the medium should be adjusted to 7.0
Isolation of rhizobium
The root sample of healthy soyabean plant were taken from agricultural field. sterilization of the root nodules
The nodules were cut from the root in such a way that around 2mm of root portion was attached to the nodules.before crushing of the nodules , the fresh nodules were taken for surface sterilization.
Isolation of rhizobium on media
The suspension prepared with crushed nodules was used for making dilution series from 10 to 10powar 10 for diluting the microorganisms number in water.
Pour plat method
The pour plat Method is based on diluting the mixed culture with liquidified nutrient agar in such a manner that the colony formed on the plate are countable . Loopful of bacteria is inoculated into tube1 to tube 10. Each tube separately in ten plates. Solidifying medium incubate at 25C° for 24 -28h in incubator.
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