Medicinal Mushroom Cultivation

Ganoderma lucidum
Lentinus edodes
Pleurotus sajor-caju
Maitake(Morchella deliciosa

1. MEDICINAL

2. GROUP :- 4 Rahul Purohit Pradeep Kamal Saurabh Deepshika Anjali Hemant Rajendra (Leader) Shubham Abhishek

3. Ganoderma lucidum Lentinus edodes Pleurotus sajor-caju Maitake ( Morchella deliciosa ) MEDICINALLY IMPORTANT MUSHROMS

4. RAW MATERIAL Vegetable waste Forest litter Straw from Wheat/Rice Wheat bran

5. Reishi ( Ganoderma lucidum ) Introduction Reishi ( Ganoderma lucidum ) is pharmacologically as well as commercially the most importat medicinal mushroom in the world. Current global trade of about 2 billion dollers ; trade in India has crossed Rs. 100 crores annually through imports from Malaysia and China.

6. Production Reishi is grown on the saw dust of th e broad-leaved trees (mango, poplar, coconut, shisham ). Sawdust, obtained from the sawmills, is amended with 20% wheat bran and is wetted to a level of 65% moisture. Calcium sulphate (gypsum) and calcium carbonate(Chalk powder) are added to get a pH of 5.5 . The mixed substrate (700 g dry wt; 2.1 kg wet) is filled in polypropylene bags the mouth of which is then plugged with cotton after putting a plastic ring exactly like wheat grain spawn pack of mushrooms in poly bags.

7. The bags are then sterilized in autoclave at 22 p.s.i . for 2 hrs. After cooling, the substrate is spawned with wheat grain or saw dusts pawn @ 3% on the dry weight basis, as it is comparatively a slow growing fungus. Spawn run(incubation) is done at 28-35 °C in the closed rooms (high carbon dioxide) and darkness. After the complete spawn run (bags white all over), which takes about 25 days,

8. polythene top is cut at the level of the substrate totally exposing the top side and proper conditions for fruiting or pinning (temp. 28 °C,1500 ppm CO2, 800 lux light, 95% RH) are provided. Once the pins have grown up enough to form the cap which is indicated by the flattenning of the whitish top of the pin head, humidity is reduced to 80% RH and more fresh air is introduced (1000 ppm CO2). Once the cap is fully formed, which is indicated by yellowing of the cap margin (which is otherwise white),temperature is lowered to 25 °C and RH is further reduced to 60% for cap thickening, reddening and maturation of the fruit bodies.

9. Full maturity is indicated, when the cap is fully reddish brown and spores are shed on the top of the cap (see the photograph).

10. Harvesting is done by the tight plucking, holding the root with one hand and pulling up with another. Scissors and knives can also be used but no residual bud is left after harvesting. One cycle of the growing takes 10-15 days. After harvesting the first flush, conditions for pinning are again switched on (i.e. 28 °C, 95%RH, 1500 ppm CO2, 800 lux light) for staring and completing the second flush.

11. One crop takes about four months. Harvested mushrooms, after washing with water, are dried at low temperature (<50 °C) in the cabinet driers, preferably at 35 °C in the dehumidifying cabinet drier. Freeze drying is, however, the best. Reishi mushroom has very high dry matter (45% i.e. 450 g dry from 1 kg fresh).

12. Lentinula edodes Lentinula edodes is a kind of wood rot fungus. In nature, it grows on dead tree trunks or stumps. Generally speaking, the C:N ratio in the substrate should be in the range from 25 to 40: 1 in the vegetative growth stage and from 40 to 73: 1 in the reproductive stage. If nitrogen source is too rich in the reproductive phase, fruiting bodies of the mushroom are usually not formed and developed .

13. The optimum temperature of spore germination is 22-26oC. The temperature for mycelial growth ranges from 5-35o C. The optimum temperature is 23-25o C. Lentinula edodes belongs to low temperature mushrooms, the initial and development temperature of fruiting body formation is in the range of 10-20o C and the optimum temperature of fructification for most varieties of the mushroom is about 15oC. Some variety can fruit in higher temperatures, e.g. 20-23o C, usually they grow faster and have a bigger and thinner cap ( pileus ), thin and long stalk ( stipe ). Their fruiting bodies are easily opened and become flat grade mushrooms, which are considered to be low quality. The optimum pH of the substrate used in making the mushroom bag/log is about 5.0-5.5.

14. Culture media and preparation : The mushroom can grow on a variety of culture media and on different agar formulations, both natural and synthetic:- Synthetic media are often expensive and time-consuming in preparation; hence they are not commonly used for routine purposes. The potato dextrose agar, or PDA, is the simplest and the most popular medium for growing the mycelium of the mushroom.

15. Four formulas in the preparation of the substrate for the cultivation of the mushroom are given here as reference. Sawdust 82%, wheat bran 16%, gypsum 1.4%, Potassium phosphate, dibasic 0.2%, and lime 0.4%. Sawdust 54%, spent coffee grounds 30%, wheat bran 15%, and gypsum 1%. Sawdust 63%, corncob powder 20%,wheat bran 15%, Calcium superphosphate1% and gypsum 1%. Sawdust 76%, wheat bran 18%, corn powder 2%, gypsum 2%, sugar 1.2% Calcium superphosphate 0.5% and urea 0.3%

16. Pleurotus sajor-caju Pleurotus sajor-caju (grey oyster mushroom) is comparable to the high temperature, required for fructification. This mushroom suitable for tropical/subtropical areas. Its cultivation is easy with relatively less complicated procedures (Chang and Miles, 2004, Kaul and Dhar , 2007).

17. . Biological nature : The temperature for growth of mycelium is 10-35oC. The optimum growing temperature of the mycelium is 23-28o. The optimum developmental temperature of the fruiting body is 18-24oC. The optimum pH of the substrate used in making the mushroom bag/bed is 6.8-8.0. The C:N ratio in the substrate is in the range of 30-60: 1. A large circulation of air and reasonable light are required for the development of the fruiting bodies. Procedure :-

18. 2) Examples of spawn substrates: Wheat grain + 1.5% gypsum or lime. Cotton seed hull 88%, wheat bran 10%, sugar 1% and gypsum 1%. Sawdust 78%, wheat bran 20%, sugar 1% and gypsum 1%. Sawdust 58%, spent coffee grounds/spent tea leaves 20%, water hyacinth/cereal straw 20%, sugar 1% and gypsum 1%. 3) Examples of cultivation substrates: Cotton seed hull 95%, gypsum 2%, lime 1% and Calciumsuperphosphate 2%. Rice straw 80%, cotton waste 18%, gypsum 1% and lime 1%. Water hyacinth 80%, cereal straw 17%, gypsum 2% and lime 1 %.

19. 4.Maitake ( Morchella deliciosa ) Materials and methods Thirty one collections of wild morels were made in Tasmania . Vegetative isolates were obtained from 21 collections and single spore isolates were obtained from 5 of these collections

20. Field sampling of wild Morels To ensure a variety of genetic material was available, morels were collected throughout Tasmania, Australia . Where possible, three ascocarps at different developmental stages, were collected from each site and given a collection number. Each ascocarp was removed with the first 15 mm of substrate attached to the base of the stipe and loosely wrapped in paper prior to insertion of each ascocarp into a separate paper bag . In the laboratory each collection was photographed, measured and described and a sporulating ascocarp selected for spore collection. To ensure that the ascocarp did not dry out too quickly and to enhance spore collection, the sporulating structure was wrapped in grease proof paper, aluminium foil and placed in a separate paper bag until spore collection was complete , or the ascocarp had dried

21. Vegetative cultures Axenic vegetative cultures were obtained from:- Malt extract agar :- (0.5 MEA; g l-1: agar, 15; malt extract, 10 ). Potato dextrose agar :- (0.5 PDA; g l-1: agar , 7.5; potato dextrose agar, 20) In 90 mm disposable Petri plates and incubated in the dark at 25°C until growth was established.

22. Single spore isolates All single spore isolates were grown from freshly germinated ascospores taken from one ascocarp . Where possible, 50 single spore isolates (SSIs) were obtained from a single ascocarp . Spores were placed in sterile distilled water, and a dilution series prepared . At each step the spore suspension was aseptically transferred to 0.5 MEA in a 90 mm Petri plate, and spread over the agar surface. Plates were incubated in the dark at 25 ºC with spore germination checked at 24 hours after inoculation and then at 12 hourly intervals. The resulting fungal colony was observed at days 10, 30, 50 and 60.

23. Precautions Keep the spawn running room dark so that spawn running will be faster. Periodically sprinkle water on sand layer to maintain the required conditions. Never spray any insecticides on the mushroom beds. In case of contamination, the contaminated block should be remove to spot well away from the house and buried in a pit or burnt. Broken pieces of the mushroom stalk, while harvesting , should not be left on the blocks. If the stalk breaks, it should be removed entirely from the bed.

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