isolation of pure culture.pptx

isolation of pure culture of mushroom
oyster mushroom
button mushroom

1. Isolation of pure culture mushroom

2. Submitting to_ Dr. Sneha J. Mistry 2 z Demonstrating by_ Patat Mayank (4011120020) Patel jay (4011120022)

3. Introduction: A mushroom is the fleshy , spore-bearing fruiting body of a fungus. The isolation of a pure culture of mushroom involves the process of separating individual fungal cells from other contaminants present in the original sample. Then growing these isolated cells into a pure culture on a nutrient medium. Introduction 3

4. Isolation: The process of strain isolation involves transferring these individual growth, or sector away from each other, so that each can be expanded and fruited to determine the best performing strains. Pure culture: A pure culture is a sample of one type of microorganism, grown in a controlled environment. Mycelium of mushroom that grows on culture media for spawn production. Introduction 4

5. Method of isolation of pure culture: Tissue culture : tissue culture is used to bring edible mushroom to pure culture so that the mushroom fungus can further be used to prepare spawn. Sub culture : sub culture means transfer of mushroom culture into fresh culture media to increase its quantity to be used for propagation. 5

6. Preparation of pure culture by Tissue culture method:

7. Instruments: Petri plates. Agar medium such as potato dextrose agar (PDA). Tweezers, needles, and spatulas. Sterilization flame of alcohol lamp. Parafilm or tape. Ethanol for burning and preparation of 70% alcohol as disinfectant. 7

8. General step in tissue culture 8

9. Procedure: Sterilize the laminar air flow. Pass UV light radiation for 15 minutes in laminar air flow. After, do all process in laminar air flow. Selection of healthy and mature fruiting body of the mushroom. Use hand gloves and cut mushroom with razor. 9

10. Procedure: Take a tissue in square shape from stip portion of mushroom. Sterile tweezer with burning lamp. Pour tissue in 0.1 Hgcl2 solution for 1 min. in petri dish. Wash it with sterile distilled water for three times in respective petri dish. 10

11. Procedure: For drying it, place tissue on blotting paper for minute. Put dried tissue in prepared PDA medium in petri dish. After, close petri dish with parafilm tape, and covered petri dish with silver coil paper. Put that petri dish in incubator under appropriate condition for 8-10 days. 11

12. Precautions : 12 Maintain sterile condition to prevent contamination. Use proper equipment. Work quickly because the longer tissue is exposed to air, the grater risk of contamination. Use a laminar air flow hood to maintain sterile environment. Monitor temperature and humidity. Properly label cultures to keep records. This Photo by Unknown Author is licensed under CC BY-SA-NC

13. Advantage of tissue culture: 13 Tissue culture allows for clonal propagation of mushroom, which ensures genetic uniformity Tissue culture can be used to produce disease-free plantlets. Tissue culture can produce a large number of plantlets in a short period of time. That is requires very little space compared to other method. Reduced the long term labor cost and increased productivity.

14. Disadvantage of tissue culture: 14 Tissue culture requires a sterile laboratory environment and high cost equipment. Tissue culture is a complex process that requires technical expertise. Limited genetic diversity. Tissue culture requires rigorous quality control measures to ensure that the cultures are free from contamination and are healthy. This method require frequent monitoring and testing.

15. Sub-culture: When tissue becomes evident growing out of the transferred mushroom pieces without contamination. Than the new isolate is sub-cultured onto new growth medium under aseptic condition. New culture are assigned unique numbers and maintained as mother culture. 15

16. Thank You 16