Need of project:
There is need of increasing nitrogen percentage in soil. Hence there is need to fix nitrogen from air by using nitrogen fixing bacteria. In vigyan Ashram Rhizobium as biofertilizer is not available. Hence we want to isolate and product make it available
Objective:
Isolation of Rhizobium
Mass multiplication of Rhizobium
Production of Rhizobium
Date: 16/07/2021
Aim: Isolation and mass multiplication of Rhizobium.
Equipment:
– Test tube, Flask, Spreader ,Sterile distilled water, Root nodules of fenugreek plant ,Yeast extract mannitol agar, petri plates.
Procedure :
Part A : Sample Collection
Leguminous roots of fenugreek plants were collected from pabal village .
Roots nodules were separated from the roots of plants.
Root nodules were primarily washed with 70% ethanol solution. And this solution was used further as a sample.
Root nodules were crushed in sterile distilled water. And is further used as stock culture.
Date: 17/07/2021
| Sr.No. | Content | Amount |
| 1. | Mannitol | 4 gm |
| 2. | Di potassium phosphate(K2PO40) | 0.2 gm |
| 3. | Magnesium sulphate(MgSo4) | 0.008 gm |
| 4. | Sodium Chloride(Nacl) | 0.4 gm |
| 5. | Yeast extract | 0.04 gm |
| 6. | Agar Agar | 10 gm |
| 7. | Calcium carbonate (CaCo3) | 1.2 gm |
| 8. | PH | |
| 9. | 1% congo red | 1 ml |
| 10. | Distilled Water | 400 ml |
Date: 18/07/2021
Part B : Isolation of rhizobium from stock culture.
20 plates, 10 test tubes and 200 ml of distilled water was sterile by using autoclave.
Yeast extract manitol agar was poured in sterile petri plates.
Date: 20/07/2021
Dilution of stock culture was prepared.
Dilution was poured on yeast extract mannitol agar plates by using spread plate techniques and 4 quadrant streaking method.
. Plates were incubated for 2-3 days at 35 degree C.
Result :
No growth is observed because by mistake UV light is turn On for 15 min on Stock culture.
Date:22/07/2021
1.Again Stock culture was prepare.
2. Dilution was poured on yeast extract mannitol agar plates by using spread plate techniques and 4 quadrant streaking method.
3. Plates were incubated for 2-3 days at 35 degree C.
Date: 24/07/2021
Result :
All isolates of rhizobium bacteria shown growth in 2 days. white, translucent , elevated colony was seen.
