isolation of pure culture of mushroom oyster mushroom button mushroom

1. Isolation of pure
culture
mushroom

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

3. Introduction:
• A mushroom is the fleshy , spore-bearin
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:
1) 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.
2) Sub culture : sub culture means transfer of
mushroom culture into fresh culture media to
increase its quantity to be used for
propagation.
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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.
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8. General step in tissue culture
Selection of a suitable source of
mushroom
Sterilization of the equipment
Extraction of tissue and transferred
onto medium
Incubation
Sub-culturing
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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.
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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.
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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.
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12. Precautions:
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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.

13. Advantage of tissue culture:
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• 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:
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• 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.
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16. Thank You
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