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mushroom Spawn preparation

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Debajit Rabha

This document provides an introduction to mushroom spawn preparation. It defines mushrooms and their life cycle. Mushrooms are nutritious and can be cultivated with low investment for profit. Spawn is the seed of mushrooms and is prepared by growing fungal mycelium in sterile grains. The process of spawn preparation involves making a pure culture, mother spawn in bottles, and commercial spawn in bags. Proper storage, transport, and precautions are needed to prevent contamination of the spawn.

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Mushrooms: Introduction and spawn preparation Presented by Debajit Rabha Asst. Prof. of Botany Pandit Deendayal Upadhyaya Adarsha Mahavidyalaya, Amjonga, Goalpara, Assam
Contents Introduction What is Mushroom? Why Mushroom? Success story Process of Mushroom cultivation What is Spawn? Preparation of Spawn Storage of spawn Transport of spawn Precautions
Introduction Mushrooms are nature's hidden treasures of nutrition The mushrooms were used as food since long back, probably from 3000 B.C. as per ancient Indian literature. Since that time, the mushrooms are being consumed in different countries like Greece, Egypt, France etc. The Greeks and Romans described mushrooms as food for the god.
What Are Mushrooms ? The mushroom is a fruiting body of microorganisms called fungi, commonly belonging to Basidiomycotina and also Ascomycotina. The mycelium, as a whole, is the non-reproductive, vegetative part of the mushroom found in soil or other organic matter. The mushroom is composed of an underground part (mycelium) and an aboveground, often edible part that is also the reproductive organ.
A labeled mushroom
Life cycle of Mushroom
Why Mushroom? Good taste and unique flavour Easy to Cultivate with short crop duration Medicinal appeal Less investment and high return Self employment (Suitable for woman) Less time and labour intensive Utilization of agricultural waste High nutritional value Suitable for small and landless farmers (space!) High market demand Spent mushroom substrate can be used for vermicomposting Asias climate suitable for mushroom cultivation
Food value Protein -Most mushrooms have a high protein content, usually around 20-30% by dry weight. Fiber -Helps lower cholesterol and is important for the digestive system. Vitamin D -Essential for the absorption of calcium. Copper -Aids in helping the body absorb oxygen and create red blood cells. Selenium -An antioxidant that helps neutralize free radicals. Mushrooms contain more selenium than any other biopropuct. Potassium -An extremely important mineral that regulates blood pressure and keeps cells functioning properly Other important minerals -Such as p, zn, and mg. Low levels of fat, calories, and sodium No cholesterol
Success story National Award Hemanta Nath, a farmer from Assam received the Progressive Mushroom Grower Award from Indian Council of Agricultural Research Directorate of Mushroom Research (ICAR DMR), Solan, Himachal Pradesh. Source: The Hill Times, 13/0922 Started from 2011 Then Oyster=10 kg/day Now Oyster=150 kg/day Milky= 10 kg/day Employs Men=4 Women=8 trademark brand 2019 Kath Fu
Hemanta Nath
Commercially cultivated mushroom in India Sl no. Common name Scientic name Growing season 1 Button Mushroom Agaricus spp. April-October Kharif 2 Milky Mushroom Calocybe spp. 3 Oyster Mushroom Pleurotus spp. Nov- Feb Rabi 4 Paddy Straw Mushroom Volvariella spp.
Commercially cultivated mushroom Oyster Mushroom
Button Mushroom
Paddy straw Mushroom
Milky Mushroom
Production of Mushroom in India
Production of Mushroom in India Button 73% Oyster 16% Paddy straw 8% Milky 3%
Mushroom cultivation Spawn preparation: Spawn is the mushroom seed. It is prepared by growing fungal mycelium in grains under sterile conditions. Composting: Compost is prepared by mixing paddy straw with number of organic materials like cow dung and inorganic fertilizers. It is kept at about 50oC for one week. Spawning: Spawn is sown on compost. Casing: Compost is covered with a thin layer of soil. It gives support to the growing mushroom, provides humidity and helps regulate the temperature.
Mushroom cultivation (continued) Pinning Mycelium starts to form little bud, which will develop into mushroom. Those little white buds are called pins. Harvesting Mushroom grow better in 15oC - 23oC. They grow 3 cm in a week which is the normal size for harvesting. In the third week the first flush mushroom can be harvested.
Major steps in Mushroom cultivation
What is Spawn? Spawn is the seed ! of mushroom. The spawn (seed of mushroom) is a pure culture of the mycelia grown on a special medium. The medium is prepared by the grains of wheat, rye, sorghum or bajra along with some ingredients.
Spawn production Spawn laboratory Spawn culture media Pure culture Preparation of mother spawn Preparation of commercial spawn Precaution
Spawn laboratory Equipment of spawn laboratory Autoclaves One boiling vessel, Laminar air flow chamber refrigerator BOD incubator pH meter Glassware Chemicals (for culture media) Non-absorbent cotton Polypropylene bags or bottles Air conditioning equipments Steel racks Exhaust fans Filters
Equipment Required Laminar air flow chamber BOD incubator Autoclave
Spawn Culture media Potato dextrose agar medium : Boil 250 g peeled potatoes in one litre of distilled water till these become soft and filter. Add 20 g dextrose and 20 g agar to the filtrate and raise volume of water to one litre. pH of the medium is maintained at 6.5. Fill about 5ml of medium in each test tube. Autoclave test tubes at 15 lb. psi for 20 to 30 minutes.
Pure culture 1. Tissue culture : especially the lower portion of the cap where the gill plate joins the stem is considered the best tissue for excision. The mushroom is cut lengthwise from cap downwards. With a flamed needle, a small piece of the internal tissue of the broken mushroom is cut and inserted into culture media bottle. The inoculated bottles are incubated at 25+1oC preferably in darkness for about 10-15 days. 2. Spore culture : Collect aseptically large sized healthy mushroom with membrane (veil) still intact, sterilize the surface, mount on a wire stand over a Petridish under sterilized glass beaker. The spores are stored under sterile condition in a refrigerator for future use. These spores can be used for direct inoculation of wheat extract agar medium or Lambert's agar medium
Pure Culture
Pure Culture
Preparation of mother spawn Soak the sorghum or wheat grains in clean water to remove chaffy and damaged grains. Cook the grains in a vessel for 30 minutes just to soften them. Take out the cooked grains and spread evenly on the platform to remove excess water. At 50% moisture level mix calcium carbonate (CaCO3) thoroughly with the cooked, dried grains @ 20 g / Kg . Fill the grains in saline bottles up to 3/4th height (approximately 300-330 g / bottle), insert a PVC ring , bold the edges of the bag down and plug the mouth tightly with non-absorbent cotton wool. The plugs are covered with aluminum foil.
Continued Arrange the bags inside an autoclave and sterilize under 20- lbs. pressure for 2 hours. Take out the bags after cooling and keep them inside the culture room and put on the UV light. After 20 minutes put off the UV light and start working in the culture room. These bottles are gently shaked on 5th and 10th day for distributing the inoculum evenly in the bottles. This spawn prepared using pure culture mycelium as inocculant is referred as mother spawn. Fully colonized mother spawn bottles can be used for inoculating commercial spawn bags after two to three weeks.
Preparation of Spawn bags
Preparation of commercial spawn Commercial spawn can be prepared in polypropylene bags (heat resistant). Normally for half and one kg spawn, the bags should be of 35 x 17.5 cm and 40 x 20 cm in size, respectively. Polypropylene bags should have double sealing at the bottom. After filling the grains a polypropylene neck (height 2 cm, dia. 4 cm) is placed near top by passing polypropylene bag through PP ring and folding back the bag (Fig. 5.6). The bags are plugged with non-absorbent cotton.
Continued These are then sterilized at 22 p.s.i. pressure for 1.5 to 2 hours. Autoclaved bags are shaked well before inoculation so that the grains absorb the water droplets accumulated inside the bags. The sterilized bags are kept on the laminar flow under . The bags are incubated under aseptic conditions using master spawn as inoculum. Ten to fifteen grams of grains of master spawn are used for inoculating one bag and one bottle of master spawn is sufficient for inoculating 25 to 30 commercial spawn bags.
Production of mother and commercial spawn
Preparation of Mother Spawn
Preparation of Commercial Spawn
Production of spawn
Spawn storage Fully colonized bags, that is, the bags in which mycelium has covered all the grains and spawn is ready to use, can be stored in cold room (+4-6属C) for 1-2 months for future use.
Temperature requirement for storage and incubation of different mushrooms (0C)
Spawn transport It is not only important to purchase good quality spawn, but it is also important to transport and store properly. The spawn is transported from one place to another in refrigerated vans or during night when temperature does not rise above 32属C. It is important that spawn bags are not exposed to heat and dust during transport.
Precautions We should prefer to use freshly prepared spawn because the mycelium is in the state of active growth. We should inspect all the bags to ensure that no bag is contaminated, that is, having green, black growth of other fungi. We should record name of the strain, when it was inoculated and on which date it was ready, etc. We may transport spawn from one place to another in refrigerated vans or during night when temperature does not rise above 32 属C. It is important that spawn bags are not exposed to heat and dust during transport. The spawn can be stored in any cool place away from dust but should not be frozen. We should also ensure that the cotton plugs are opened only when the spawn is to be used. During spawning if we notice some bag with contamination, it should be removed and hands sterilised before continuing with the spawning.
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mushroom Spawn preparation

  • 1. Mushrooms: Introduction and spawn preparation Presented by Debajit Rabha Asst. Prof. of Botany Pandit Deendayal Upadhyaya Adarsha Mahavidyalaya, Amjonga, Goalpara, Assam
  • 2. Contents Introduction What is Mushroom? Why Mushroom? Success story Process of Mushroom cultivation What is Spawn? Preparation of Spawn Storage of spawn Transport of spawn Precautions
  • 3. Introduction Mushrooms are nature's hidden treasures of nutrition The mushrooms were used as food since long back, probably from 3000 B.C. as per ancient Indian literature. Since that time, the mushrooms are being consumed in different countries like Greece, Egypt, France etc. The Greeks and Romans described mushrooms as food for the god.
  • 4. What Are Mushrooms ? The mushroom is a fruiting body of microorganisms called fungi, commonly belonging to Basidiomycotina and also Ascomycotina. The mycelium, as a whole, is the non-reproductive, vegetative part of the mushroom found in soil or other organic matter. The mushroom is composed of an underground part (mycelium) and an aboveground, often edible part that is also the reproductive organ.
  • 6. Life cycle of Mushroom
  • 7. Why Mushroom? Good taste and unique flavour Easy to Cultivate with short crop duration Medicinal appeal Less investment and high return Self employment (Suitable for woman) Less time and labour intensive Utilization of agricultural waste High nutritional value Suitable for small and landless farmers (space!) High market demand Spent mushroom substrate can be used for vermicomposting Asias climate suitable for mushroom cultivation
  • 8. Food value Protein -Most mushrooms have a high protein content, usually around 20-30% by dry weight. Fiber -Helps lower cholesterol and is important for the digestive system. Vitamin D -Essential for the absorption of calcium. Copper -Aids in helping the body absorb oxygen and create red blood cells. Selenium -An antioxidant that helps neutralize free radicals. Mushrooms contain more selenium than any other biopropuct. Potassium -An extremely important mineral that regulates blood pressure and keeps cells functioning properly Other important minerals -Such as p, zn, and mg. Low levels of fat, calories, and sodium No cholesterol
  • 9. Success story National Award Hemanta Nath, a farmer from Assam received the Progressive Mushroom Grower Award from Indian Council of Agricultural Research Directorate of Mushroom Research (ICAR DMR), Solan, Himachal Pradesh. Source: The Hill Times, 13/0922 Started from 2011 Then Oyster=10 kg/day Now Oyster=150 kg/day Milky= 10 kg/day Employs Men=4 Women=8 trademark brand 2019 Kath Fu
  • 11. Commercially cultivated mushroom in India Sl no. Common name Scientic name Growing season 1 Button Mushroom Agaricus spp. April-October Kharif 2 Milky Mushroom Calocybe spp. 3 Oyster Mushroom Pleurotus spp. Nov- Feb Rabi 4 Paddy Straw Mushroom Volvariella spp.
  • 17. Production of Mushroom in India Button 73% Oyster 16% Paddy straw 8% Milky 3%
  • 18. Mushroom cultivation Spawn preparation: Spawn is the mushroom seed. It is prepared by growing fungal mycelium in grains under sterile conditions. Composting: Compost is prepared by mixing paddy straw with number of organic materials like cow dung and inorganic fertilizers. It is kept at about 50oC for one week. Spawning: Spawn is sown on compost. Casing: Compost is covered with a thin layer of soil. It gives support to the growing mushroom, provides humidity and helps regulate the temperature.
  • 19. Mushroom cultivation (continued) Pinning Mycelium starts to form little bud, which will develop into mushroom. Those little white buds are called pins. Harvesting Mushroom grow better in 15oC - 23oC. They grow 3 cm in a week which is the normal size for harvesting. In the third week the first flush mushroom can be harvested.
  • 20. Major steps in Mushroom cultivation
  • 21. What is Spawn? Spawn is the seed ! of mushroom. The spawn (seed of mushroom) is a pure culture of the mycelia grown on a special medium. The medium is prepared by the grains of wheat, rye, sorghum or bajra along with some ingredients.
  • 22. Spawn production Spawn laboratory Spawn culture media Pure culture Preparation of mother spawn Preparation of commercial spawn Precaution
  • 23. Spawn laboratory Equipment of spawn laboratory Autoclaves One boiling vessel, Laminar air flow chamber refrigerator BOD incubator pH meter Glassware Chemicals (for culture media) Non-absorbent cotton Polypropylene bags or bottles Air conditioning equipments Steel racks Exhaust fans Filters
  • 24. Equipment Required Laminar air flow chamber BOD incubator Autoclave
  • 25. Spawn Culture media Potato dextrose agar medium : Boil 250 g peeled potatoes in one litre of distilled water till these become soft and filter. Add 20 g dextrose and 20 g agar to the filtrate and raise volume of water to one litre. pH of the medium is maintained at 6.5. Fill about 5ml of medium in each test tube. Autoclave test tubes at 15 lb. psi for 20 to 30 minutes.
  • 26. Pure culture 1. Tissue culture : especially the lower portion of the cap where the gill plate joins the stem is considered the best tissue for excision. The mushroom is cut lengthwise from cap downwards. With a flamed needle, a small piece of the internal tissue of the broken mushroom is cut and inserted into culture media bottle. The inoculated bottles are incubated at 25+1oC preferably in darkness for about 10-15 days. 2. Spore culture : Collect aseptically large sized healthy mushroom with membrane (veil) still intact, sterilize the surface, mount on a wire stand over a Petridish under sterilized glass beaker. The spores are stored under sterile condition in a refrigerator for future use. These spores can be used for direct inoculation of wheat extract agar medium or Lambert's agar medium
  • 29. Preparation of mother spawn Soak the sorghum or wheat grains in clean water to remove chaffy and damaged grains. Cook the grains in a vessel for 30 minutes just to soften them. Take out the cooked grains and spread evenly on the platform to remove excess water. At 50% moisture level mix calcium carbonate (CaCO3) thoroughly with the cooked, dried grains @ 20 g / Kg . Fill the grains in saline bottles up to 3/4th height (approximately 300-330 g / bottle), insert a PVC ring , bold the edges of the bag down and plug the mouth tightly with non-absorbent cotton wool. The plugs are covered with aluminum foil.
  • 30. Continued Arrange the bags inside an autoclave and sterilize under 20- lbs. pressure for 2 hours. Take out the bags after cooling and keep them inside the culture room and put on the UV light. After 20 minutes put off the UV light and start working in the culture room. These bottles are gently shaked on 5th and 10th day for distributing the inoculum evenly in the bottles. This spawn prepared using pure culture mycelium as inocculant is referred as mother spawn. Fully colonized mother spawn bottles can be used for inoculating commercial spawn bags after two to three weeks.
  • 32. Preparation of commercial spawn Commercial spawn can be prepared in polypropylene bags (heat resistant). Normally for half and one kg spawn, the bags should be of 35 x 17.5 cm and 40 x 20 cm in size, respectively. Polypropylene bags should have double sealing at the bottom. After filling the grains a polypropylene neck (height 2 cm, dia. 4 cm) is placed near top by passing polypropylene bag through PP ring and folding back the bag (Fig. 5.6). The bags are plugged with non-absorbent cotton.
  • 33. Continued These are then sterilized at 22 p.s.i. pressure for 1.5 to 2 hours. Autoclaved bags are shaked well before inoculation so that the grains absorb the water droplets accumulated inside the bags. The sterilized bags are kept on the laminar flow under . The bags are incubated under aseptic conditions using master spawn as inoculum. Ten to fifteen grams of grains of master spawn are used for inoculating one bag and one bottle of master spawn is sufficient for inoculating 25 to 30 commercial spawn bags.
  • 34. Production of mother and commercial spawn
  • 38. Spawn storage Fully colonized bags, that is, the bags in which mycelium has covered all the grains and spawn is ready to use, can be stored in cold room (+4-6属C) for 1-2 months for future use.
  • 39. Temperature requirement for storage and incubation of different mushrooms (0C)
  • 40. Spawn transport It is not only important to purchase good quality spawn, but it is also important to transport and store properly. The spawn is transported from one place to another in refrigerated vans or during night when temperature does not rise above 32属C. It is important that spawn bags are not exposed to heat and dust during transport.
  • 41. Precautions We should prefer to use freshly prepared spawn because the mycelium is in the state of active growth. We should inspect all the bags to ensure that no bag is contaminated, that is, having green, black growth of other fungi. We should record name of the strain, when it was inoculated and on which date it was ready, etc. We may transport spawn from one place to another in refrigerated vans or during night when temperature does not rise above 32 属C. It is important that spawn bags are not exposed to heat and dust during transport. The spawn can be stored in any cool place away from dust but should not be frozen. We should also ensure that the cotton plugs are opened only when the spawn is to be used. During spawning if we notice some bag with contamination, it should be removed and hands sterilised before continuing with the spawning.