FREE INFORMATION GUIDE TO BASIC MUSHROOM CULTIVATION
Just follow step by step guidelines if you do not have time for hands on training. This will give you idea on how to produce spawn, fruiting bags making, pasteurization, inoculation, planting, harvesting and marketing.
Tissue Culture:Materials Needed:
Potatoes: 200 gr. Dextrose: 20 gr. Agar powder: 20 gr. Water: 1 liter. Cotton (gauze)
Note: Inspect potatoes for spots or rot. Purchase dextrose and Agar of commercial grade.
Wash thoroughly and cut potatoes into one-centimeter cubes; leave on or peel off the skin.
Clean small flat bottles (small whiskey bottles as a container can be used).Place potatoes in one liter of water. Simmer for 15 - 20 minutes.Remove potatoes & keep the broth as clear as possible.
Add water to broth to reach one liter of liquid PDABring water to stove. Add dextrose followed by agar. Slowly stir continuously with minimum speed until completely dissolved.Pour liquid PDA in bottle until you reach 5 - 10 mm high.Plug bottle with cotton.Place bottles in autoclave at 121 degrees Centigrade for 20 - 30 minutes to ensure complete sterilization.
Let cool down to around 37 degrees Centigrade. After PDA medium is settled in bottle, transfer all bottles to clean shelf in the clean room.
Check for contamination (contamination can be seen when dark spots or lines occur).
Selecting Tissue CultureMaterials Needed:
Special needle (insulated handle)
Alcohol lamp
Alcohol
Cotton (gauze)
Ultra Violet lamp
Matches or lighter
Bottles with PDA
Laminar flow cabinet (or protected environment)
Select a strong mushroom for culture.
Healthy.
Not too mature, not too young.
Not too humid (at least 2-3 hours after watering)
With a stiff stalk
Make sure it is clean and far from any contaminated mushroom.
Clean the room, all necessary tools, inside and outside the laminar flow cabinet with alcohol. Transfer PDA bottles and necessary tools into the chamber.
Place all cleaned materials inside laminar flow. Turn on UV lamp and laminar flow. After 10-15 minutes, turn off UV lamp but leave laminar flow for the duration of the operation.
Clean both hands and bottles with alcohol and insert hands into the cabinet.
Hold needle with 2 fingers in a 45 degree angle, flame needle to disinfect until the needle turns red.
Make sure it does not touch any surface after flaming.
While needle cools down (15-20 seconds - hold needle not to touch anything or place it on the clean surface of a glass).
Using other fingers, tear mushroom lengthwise (DO NOT use knife to cut).With the needle, cut a small piece (2 mm x 2 mm) of fleshy tissue from inside the mushroom (in the middle between the cap and the stalk).
Make sure that it is clean and did not touch the outside of the mushroom.Flame around the mouth of the bottle. Using other fingers, remove cotton plug of PDA bottle in front of flame to secure against contamination.
Insert the needle in the bottle and inoculate by placing small piece of cut mushroom in the middle of the PDA’s surface. Make sure the piece of mushroom does not touch anything before entering the PDA bottle.
Close bottle immediately near the flame with cotton plug.
Note: the bottom of the bottle should always be lower than the mouth of the bottle and the mouth of the bottle should remain near the flame at all times.
Label bottles and indicate:
Date, type of mushroom, mother spawn #.Whether from tissue culture or PDA to PDA, from the time of incubation to full growth mycelium will take about 10 - 15 days. (Depending on species).Keep PDA bottles with mycelium on clean shelf.Check infection by other fungi in the bottle everyday. Also check growth rate.
After mycelium covers whole PDA medium, keep mature mycelium in cool place or in the refrigerator in the vegetables section.Check for contamination. Separate contaminated bottles. Transfer contaminated bottles to clean.
MULTIPLYING SPAWN ON SORGHUM SEEDS Materials Needed:
Sweet Sorghum seeds
Bottles (flask type)
Cotton (gauze)
Paper squares 7 cm x 7 cm
Rubber bands
Alcohol lamp
Alcohol bottle
Grains must:
Have been recently harvested
Contain few broken kernels
Little contamination
No fungi, no insects
No more than 12% humidity
Soak sorghum for one night; 2 liters of water per 1 kg of grain. Wash and strain sorghum seeds to remove all water. Steam sorghum seeds for 30-45 minutes to soften grains and cook about 25%.
Drain water and spread sorghum seeds to cool down and decrease moisture.
Fill ¾ of bottle with sorghum seeds.Carefully prepare cotton plugTightly plug mouth of bottle with cotton and leave out for ventilation.Transfer all prepared bottles to the sterilization chamber.
Close chamber. Fire-up burner or stove to heat chamber. Make sure to release all air from the chamber before starting. Keep pressure in the chamber at 15 lb./sq.inch. or 121 degrees Celsius for 30 minutes for small chambers and 45 minutes for medium chambers.
Let bottles cool down.Transfer bottles to a clean and cool place. Bottles must be cleaned and well prepared.Prepare the well verified PDA bottles Clean laminar flow chamber using alcohol.
Transfer PDA, sorghum seed bottles, paper and rubber bands in laminar flow chamber. Light UV lamp for 10 - 15 minutes before starting. Place needle in alcohol.
Turn off UV. Clean both hands with alcohol and insert hands into the chamber.
Using 2 fingers, take out needle, pass through fire as to burn alcohol, and disinfect needle. Make sure the needle turns red.
After the needle cooled down to normal state, use needle to cut small square (5mm x 5mm) of PDA with mycelium (white color).
Close bottle immediately. Remain near flame at all times. Using other hand flame around the mouth and shoulder of the sorghum seed bottle.Using other fingers, open spawn bottle near flame to avoid contamination. Insert needle and inoculate sorghum seeds with PDA mycelium by placing small square piece in the middle of the bottle. Make sure the PDA mycelium does not touch anything before entering the sorghum seeds bottle.
Note: The mouth of the bottle should be near the flame. The mouth should remain higher than the bottom part at all times. Do not touch mouth of bottle with piece of PDA.
Close bottle immediately.Place square paper over cotton and tie with plastic neck or rubber band. Label inoculated sorghum bottles writing: Date, Spawn no., ref., and inoculation time.
Note: It takes about 10 - 15 days to get full-grown sorghum grain mycelium, depending on the species.
Keep mature sorghum seeds in a cool place or in the vegetable compartment of the refrigerator. Check for infection regularly.Remove contaminated bottles. Transfer contaminated bottles to cleaning site. Clean bottles as normal glassware.
Observe and collect data. Take notes to draw conclusions.
Note: A loss of about 3% is to be expected.
PRODUCING SUBSTRATE BAGS
Materials Needed:
Plastic bags (polypropylene 8” x 16”)
Plastic necks (about 5 cm diameter)
Cotton plugs Cover filters (square piece of filter paper)
Rubber bands Sawdust (cover top of substrate pile with rice sacks to maintain humidity)
Rice bran
Calcium sulfate
Calcium carbonate
Magnesium sulfate
Substrate is the material used to grow mushrooms. This material or substrate is a mixture of all ingredients or “food” necessary for mushrooms to develop. Although sawdust is the most common and easy to use basic material for making mushroom substrate bags, other alternate and sometimes lower cost materials can be used. For example, in Asia, because of intense rice cultivation, rice straw can be used since straw is readily available in most rural areas. Because of its lower cost (and local availability) it may be better suited as a substrate than sawdust. Furthermore, rice straw generally generates higher yield and better quality mushrooms; both texture and taste of mushrooms are improved when using straw instead of sawdust. Nevertheless, straw needs to be prepared before use requiring harder work, and fermentation for a period of 9 - 12 days.
Preparing rice straw as substrate:
Put straw in grinder to reduce its size. Soak paddy straw 100 Kg. Ferment for 3 days. Turn over the pile, then mix with 2% lime and ferment it again 3 days. Turn over the pile again, mix with 0.2% magnesium sulfate, and ferment 3 more days. The last turn over makes the straw readily composted for using as substrate. Check moisture and for a composted smell. The substrate is ready for packing. Then pack in size 8” x 16” PP. Bags.
Substrate preparation:
100 kg Sawdust Add to sawdust 5 kg Rice bran 2 kg Calcium sulfate 1 kg Calcium carbonate 0.2 kg Magnesium sulfate 0-1 kg Sugar
Note: Substrate recipe should serve as a reference. Recipe can be changed by adding some rice flour, sticky rice flour, corn flour, cassava peels, cotton waste, Soya-bean residue, and other nutritious agricultural waste. In cool climates, it is possible to use additives or complementing materials up to 20%. Beware: for hot climatic zones, do not use more than 7.5% additives. (If rice straw, recipe needs to be modified as above mentioned) Weigh all components using scale.Mix well all ingredients in mixer or manually using shovels or paddles.Add water to keep moisture content between 65-75 %. Make sure all ingredients are well mixed and that there are no lumps.Sitting on floor, fill plastic bags with substrate using small shovel.
Compact substrate by hitting bags with empty bottle, hand.Or,use compacting machine. Place plastic ring on bag.Pull out top of bag through plastic neck. Fold-down on plastic ring.Tie with rubber band.Introduce stick with pointed head through plastic neck of bag to make hole almost to the bottom of the bag; DO NOT TOUCH BOTTOM of bag. Check weight of bags (should be between 800-1000 grams per bag).Prepare plastic caps to close bags by adding cotton.