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Present Situation and Prospect of Mushroom Cultivation in Nepal

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Present Situation and Prospect of Mushroom Cultivation in Nepal Lecture 3 Effect of genetic factors, temperature, light, humidity, medium on cultivation of mushrooms. Techniques of Commercial cultivation of some important mushrooms, Single spore isolation/pure culture and spawn production techniques, Present situation and prospect of mushroom cultivation in Nepal. Mitesh Shrestha Genetic Factors • For breeding work in edible species it is desirable to know how fruiting is controlled genetically as well as nutritionally and physiologically. Genetic Factors Temperature • The conditions that are favorable for reproduction are always less favorable for growth. • An increase in temperature generally increases enzymatic activity. In that part of the growth curve that is linear, the growth rates approximately double for each 100C increase in temperature (i.e., the Q10 value is 2). • High temperatures inactivate enzymes with a resulting effect on metabolism and, consequently, on growth. The failure of a fungus to grow at high temperature may be the result of inability to synthesize a needed vitamin, for example. Effect of Temperature, Light and Humidity Temperature (in oC) <0 = no growth >0 – 10 = Very very slow growth >10 – 15 = Slow growth .15 – 22 = Good growth >22 – 25 = very fast growth >25 = may case harm to mycelia and its growth Pholiota adiposa • Mycelial growth was found to occur over the range of 5 to 33°C; whereas fruiting occurred only from 13 to 24°C. The optimal temperature for mycelial growth was 27°C and for fruiting body formation 18 to 21°C. Light • The growth of most fungi is not sensitive to light, although strong light may inhibit or even kill (possibly a temperature effect). • One exception to this latter statement concerning S. commune is of interest. Bromberg and Schwalb have demonstrated that light is required in S. commune for the formation of primordia and also for the early stages of fruiting body development in which short cylindrical stipes with terminal apical pits are formed. • It has been reported that this inhibition by strong light may be overcome by the addition of natural materials, such as yeast extract, to the medium. A possible interpretation for this phenomenon is that the light may have destroyed certain vitamins, which are then replaced by those present in the natural material. • Because light is actually inhibitory to the development of primordia and affects stipe elongation and pileus expansion of the button mushroom (Agaricus bisporus), this mushroom is grown in caves, tunnels, or mushroom houses in the dark. Light (sun light / electrical light / no light ) • Intense light • Semi light • Dark • Open (without shade) – In forest – In land / field • Shady (in forest, Hut / house / covered area) – semi shady – Shady • Dark • Semi dark Humidity • It is generally recognized that most fungi require high moisture levels. • A relative humidity of 95 to 100% and a moisture content of the substrate between 50 and 75% support maximum growth of most Basidiomycetes; but, of course, there are fungi adapted to lower moisture concentrations. • The extreme is the germination of powdery mildew spores that occurs at 0% relative humidity. • Another exception is the dry rot fungus, Serpula lacrymans, whose mycelial strands can grow through substrates lacking moisture as a result of translocation of nutrients farther back in the hyphae and by the formation of “metabolic water.” Humidity • <0 = no symptom of growth • >0 – 15 = very very slow growth • >15 – 30 = very slow growth • > 30 – 50 = medium growth • >50 – 70 = Good grwoth • > 70- 80 = Fast growth • >80 – 100 = very fast growth but may cause harm Aeration • The components of the air that are of greater importance to most fungi are oxygen and carbon dioxide. Most fungi are obligate aerobes, but many will also grow in reduced levels of oxygen. • The mushrooms of Basidiomycetes may be malformed in the presence of too much respiratory carbon dioxide - a fact that emphasizes the need for proper ventilation in mushroom growing houses. • The effect of high concentrations of carbon dioxide on the development of fruiting bodies was observed with elongation of the stipe. • The influence of carbon dioxide on fruiting body development of A. bisporus was extensively studied and reported as abnormal pileus formation as well as extensive stipe elongation. • The vegetative growth of A. bisporus has been shown to require low levels of carbon dioxide with an optimum concentration in the range of 0.1 to 0.5% reported by San Antonio and Thomas pH • It is well known from experimental studies that the optimal pH values for fruiting may differ from those for growth. It is also known that species differ in their optimal pH values for fruiting. • During the course of an experiment the pH value of the medium may change because the fungus has produced metabolites, e.g., organic acids, that affect the hydrogen ion concentration. • Media that are strongly buffered or that are periodically neutralized by the addition of base or acid may not fruit because the pH value required for the metabolic reactions necessary for fruiting is not reached. pH Media • Carbon – Carbon sources provide for both the structural and energy requirements of the fungal cell. The fungi are quite versatile in utilization of carbon compounds. There are fungal species that utilize various polysaccharides, monosaccharides, organic acids, amino acids, certain alcohols, polycyclic compounds, and natural products such as lignin and cellulose as carbon sources. • Nitrogen – Chitin, a polysaccharide of common occurrence in the cell walls of many fungi, also contains nitrogen. • Minerals – Phosphorous, Potassium, Magensium etc. • Vitamins – Thiamine, Biotin, Nicotinic Acid, Pantothenic Acid etc. Six steps for growing Mushroom • Making Mushroom Compost • Finishing the Compost • Spawning • Casing • Pinning • Cropping Materials necessary for cultivation Water for cleaning or washing Autoclave, incubators, Inoculating chamber, Laminar Air flows Spirit lamp. Rectified spirit, lighter Cotton non absorbent Inoculating needle Blades Test tubes petri dishes. Paddy strwa, Wood logs, Eledtric boarer Wax Wheat, Chemicals Plastic bags, wooden trays, Rubber bands Chemicals and materials necessary PDA (for slants) Boil and pour into test tubes and Strelize 1. Potato raw 200 g. 2. Dextrose sugar 15 g. 3. Agar Agar 20g. Or 25 grms of PDA in 1000 ml of water For making spawn (growing mycelium on wheat seeds) 1. Wheat seeds 20 kg. 2. Lime 30 – 25 gr. Wash and soak wheat grains for one day, mix lime, fill the bags, plug them, sterelixe Preparation of PDA medium for mushrooms Making spawn for cultivation Culture tubes of mushrooms Making spawn for cultivation Substrate for growing Agaricus bisporus Cow dung . Horse manure, droppings Paddy straw, Chopped straws Chemicals – Ammonium sulphate, Potassium, Urea Calcium carbonate Water Temperature thermometer Wooden wracks, trays Process For paddy straw mushrooms 1. Select good straw 2. Chop upper and lower parts and throw 3. Cop whole part in to 3- 4 “ 4. Wash throughly by water 5. Soak in water for one day 6. Wash throughly by water 7. Keep in shady place for 2 – 3 hours, drain water 8. Pack up straw in plastic bags with grains of mushroom spawn 9. Seal with rubber bads Chemicals necessary For composting ( For One ton farm yard manure / organic manure/ Dungs + Paddy straw mix) 1. Ammonium sulphate 5.6 kg 2. Super phosphate 5.6 kg - At first and second 3. Lime 20.0 kg – At first and second 4. Gypsum 16 kg - At first and second 5. Copper sulphate 2.0 Oz. – At first Process Mix in the stacks / heaps / debris breaking in definite ratio in each turnover after one week Turn over the heaps sprinkling the water after each three days Process For compost + straw mushrooms 1. Make heaps of dungs (one ton each) 2. Make dust of dung / coarse garined dungs 3. Mix with small pices of straw 2-3 “ (clean) 4. Make heaps / stacks 5. Sprinkle water regularly by turning 6. Add chemical fertilizers 7. Keep in shady place and turn the compost after 2 – 3 days 8. Do not let exceed the temperature over 110 –120c Spawning and Casing the trays 1. Fill trays with well made manure 2. Make ½” hole with glass rods 3. insert spawn grains 4. Cover slightly with manure 5. Cover the trays with moist papers 6. See development of mycelium after 15 – 20 days 7. Remove the paper 8. Case the trays with sterilized soli Stages in Growing Agaricus bisporus Pleurotus eryngii Flammulina velutipes Materials necessary for Pleurotus cultivation : sGo] Rofp v]tL k/fn M % lsnf] Paddy straw e':;f sf6\g] s}FrL M ! Uff]6f Chopping tool 8|||«|||\|d jf kmf]l; / kf]6f;L M ! Uf]f6f Space, Drum, vessel, Knfl:6s y}nf -!@ !^ OGr_ M % j6f Plastic bag × Knfl:6s l;6 M ! ld6/ Plastic sheet Knfl:6s af6f M ! Uf]f6f Plastic vessel bfp/f M !) lsnf] Rofpsf] lapm M ! af]tn Spawn / seed of mushroom 8f]/L M $ 6's|f Rubber band / Rope k|ljlw k/fnsf] u'0f:t/ M ;kmf . xl/of]kg gePsf] . k/fn 6'qmf kfg]{ M @ b]lv # OGr . k/fnsf] 6'qmf tf}ng] M % s]hL k|lt af]tn -@%) u|f aLpm _ k/fn lehfpg] M @ b]lv # 306f . k/fn w'g] M # b]lv $ k6s . kfgL tsf{pg] M % b]lv ^ 306f jf ! /ft k/fn akmfpg] M ! b]lv !=% 306f - /f]u, ls/f zq' lgoGq0fsf nflu_ k/fn lr:ofpg] M % b]lv ^ 306f Mushroom huts for cultivation Substrate for growing Pleurotus ostreatus & Lentinus sajor-caju Paddy straw, Chopped straws Chemicals – Ammonium sulphate, Potassium, Urea Calcium carbonate Water Temperature thermometer Plastic bags or other containers Agaricus bisporus • In Phase I of the process (outdoor composting), locally available raw materials are arranged into piles which are periodically turned and watered. The initial breakdown of the raw ingredients by microorganisms takes place in Phase I.
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