PRASHANT KUMAR SHARMA Biotech Lab, Training and Demonstration Centre, AMBIKAPUR (CHHATTISGARH) INDIA

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Asian J. Bio Sci. (2007) Vol. 2 No. 2 : (131-136) 131 Effect chemical supplementation on the and biological efficiency Pleurotus sajor caju grown in three different lignocellulosic wastes in Chhattisgarh, India PRASHANT KUMAR SHARMA Biotech Lab, Training and Demonstration Centre, AMBIKAPUR (CHHATTISGARH) INDIA (Accepted : August, 2007) In recent years the mushroom technology has traveled far a head. The domestication various mushroom species has been tried globally, many which are now commercially cultivated for food as well as medicinal purposes for an amateur and pressional cultivator. The production mushroom has become important factor, which does not promote the growers. The important priority pression is to maximize the production mushroom by using various techniques. Different concentration carbon sources i.e. lactose (1.0%, 2.0% and 3.0%), nitrogen sources i.e. peptone (1.0%, 2.0% and 3.0%), inorganic chemicals i.e. Mgso 4 (1.0%, 2.0% and 3.0%), vitamin sources i.e. vit. B-complex (100, 200 and 300 mg/l) and growth promoter i.e. EDTA (0.1%, 0.2% and 0.3%) were screened to determine the most suitable concentration for better Pleurotus sajor caju. Three lignocellulosic agricultural waste material i.e. wheat straw, paddy straw and leman grass straw used for cultivation Pleurotus sajor caju It was found that carbon source and inorganic chemical gave maximum and biological efficiency in 2.0% concentration in wheat straw. On that basis lactose and Mgso 4 were proved superior carbon source and inorganic chemical, respectively. peptone, the semisolid protein as nitrogen source gave maximum and biological efficiency in 3.0% concentration in lemon grass straw while in case growth promoter it was observed maximum in 0.2% concentration in wheat straw. vitamin B-complex gave maximum and biological efficiency in 300-mg/l concentrations in wheat straw, among the three lignocellulosic agricultural wastes material used as substrate, best growth and high Pleurotus sajor caju was obtained in wheat straw. Key words : Chemical supplement, Biological efficiency, Lignocellulosic waste. INTRODUCTION The cultivation edible fungi has been accepted as the easiest on farm biotechnology for pritable removal various lignocellulosic agricultural, industrial and forestry by-products, specially in the developing countries, although about 200 species edible fungi have been reported from India, only three viz., Pleurotus spp., Agraricus bisporus and Volvariella spp. are preferred for commercial cultivation and Pleurotus spp. alone constitutes about 65% the total mushrooms production in the country (Munjal, 1982). These species mushrooms belong to the class basidiomycotina and exhibit two distinct phases in their life cycle, a vegetative phase called hyphae which branch and extend into mycelium and a reproductive phase characterized by the development a fleshy sporophore referred to as mushroom fruit body. It is this structure, which is prized for its high nutritive value and culinary properties (Mehta, 1990). Mushrooms are a class heterotrophic fungi and due to the absence chlorophyll in their cells they completely depend on the substrate for all their nutritional requirement carbon, water, nitrogen and minerals. In any cultivation programme on mushroom the primary requiste is preparing a suitable substrate. Pleurotus sajor caju can grow on a variety fresh lignocellulosic residues requiring very little pretreatment (Bano et al., 1982). Mushrooms are used as delicious flavored food and having nutritional value between meat and vegetables, mushrooms are rich in protein, vitamin and minerals. It is low caloric food with very high potassium, sodium ratio and without starch as well as cholesterol. Vast quantities renewable lignocelluloses wastes are generated every year in developing countries like India with economics, which are basically agricultural. However mushroom spp. have been reported to grow and successfully on many plant wastes. In special reference chhattisgarh 36.4 lakh ton agricultural waste material left after animal consumptions can be used as substrates for mushroom production.

132 SHARMA Present communication, different concentrations carbon source (lactose), nitrogen source (peptone), inorganic chemicals (Mgso 4 ), growth promoters (EDTA) and vitamin source (B- Complex) were screened to determine the most suitable concentration for better, biological efficiency and suitable substrates Pleurotus sajor caju. The aim various different chemical supplementations is to get maximum production fruiting bodies but it is an intricate problem. Moreover, the knowledge, shell help to plan and prepare for the crop and equip one with a physiological tool to control the quality, quantity, timing and other characters the mushroom cultivation. MATERIAL AND METHODS This work was carried out in Biotech lab training and demonstration center, Ambikapur, chhattisgarh during april 2006 to december 2006, parental strains Pleurotus sajor caju were provided by mushroom biotechnology lab, india gandhi agriculture university, raipur, Chhattisgarh. The cultures were maintained on malt extract agar medium with regular sub-cultures at monthly intervals, spawn was prepared on wheat grain following standard procedure (Upadhyay et al., 1997). 15 days old spawn was used for experimentation. Different concentration carbon sources i.e. lactose (1.0%, 2.0% and 3.0%), nitrogen sources i.e. peptone (1.0%, 2.0% and 3.0%), inorganic chemicals i.e. Mgso 4 (1.0%, 2.0% and 3.0%), vitamin sources i.e. vitamin B-complex (100, 200 and 300 mg/l) and growth promoter i.e. ethylene diamine tetracetic acid (0.1%, 0.2% and 0.3%) were selected for the research purpose. In any mushroom cultivation programme the primary requisite in preparing a suitable substrate. Pleurotus sajor caju can grow on a variety fresh lignocellulosic residues requiring very little pretreatment (Yadav et al., 1998), Three lignocellulosic agricultural waste i.e. wheat straw, paddy straw and lemongrass straw used for cultivation Pleurotus sajor caju. straw, paddy straw and lemon grass straw was water socked over night in 1% bavistin and 2% formaldehyde solution, at the following day, the substrate was spread clean floor to drain f excess water. Polythene begs was prepared by layering methods. All the carbon sources i.e. lactose (1.0%, 2.0% and 3.0%), nitrogen sources i.e. peptone (1.0%, 2.0% and 3.0%), inorganic chemicals i.e. Feso 4 and Mgso 4 (1.0%, 2.0% and 3.0%) were supplemented into the substrate just before the spawning in the case growth promoters i.e. ethylene diamine tetracetic acid (0.1%, 0.2% and 0.3%) & vitamin source i.e. vitamin B-complex (100, 200 and 300 mg/l) 20 ml solution different concentrations were sprayed, respectively on the surface begs and at the time pinhead and after the fruiting bodies, the bags were incubated in cultivation room at 25 0 C-30 0 C for spawn after completion spawn, time pinhead, and biological efficiency were separately recorded for each treatment at the time each flush. After cultivation Pleurotus sajor caju the effect chemical supplementation the fruiting bodies is dried in sunlight and than prepared powder form and biochemical analysis was done and found that biochemical content Plurotus sajor caju. RESULTS AND DISCUSSION Pleurotus sajor caju has been recognized as a highly potential converter cheap cellulosic material in to valuble protein at a very nominal cost. The results obtained during the present investigation are presented in the Table 1 to 6 and photographs. All the sets supplemented with carbon source (Lactose) took shorter time for spawn and primordial than control (16 days), the and biological efficiency was observed maximum in carbon source (lactose 2%) 940 grams, 94.0% in wheat straw, compared with control 650 gram and 65.0% biological efficiency. Minimum was recorded in carbon source i.e. 580 gram in lactose 3% concentration in lemon grass straw. These are evidences that increase in carbohydrate beyond an optimum point results in an Absolute as well as relative decrease in growth fungus, (H.S.Sohi 1992), In comparison to lactose for different straw lactose 3% lemon grass straw (580 gram) was proved to be less effective to others. The similar findings were also reported by Singh (2005). The sets treated with peptone took different time for spawn, wheat straw (13 days), paddy straw (12 days) and lemon grass (16 days) compared with control (17 days), they showed variation in day harvest (table-2), it was observed maximum in peptone 3% concentration lemon grass straw (29 days). The increasing concentrations caused positive effect on and biological efficiency with in a certain limits. It was observed the maximum in peptone 3% concentration in lemon grass straw (900 gram and 90.0 %). This was due to peptone that serves additional available nitrogen to fungus and thus stimulates fungal growth.it also helped in maintaining high cellulase activity and cell mass synthesis, various workers have

EFFECT OF CHEMICAL SUPPLEMENTATION OF P. SAJOR CAJU 133 Table 1 : Effect different concentration carbon source (Lactose) on and Biological efficiency Lactose-1% 14 18 23 4 890 89.0 Lactose-2% 14 18 23 4 940 94.0 Lactose-3% 14 20 25 4 770 77.0 Lactose-1% 13 20 26 4 772 77.2 Lactose-2% 13 20 26 4 770 77.0 Lactose-3% 13 19 25 4 695 69.5 Lactose-1% 18 24 30 4 661 66.1 Lactose-2% 17 24 30 4 670 67.0 Grass Lactose-3% 18 24 29 4 580 58.0 Control 16 20 26 4 650 65.0 Fig. 1 : Steps involve cultivation Pleurotus sajor caju. also reported that addition organic source nitrogen enhances the Pleurotus sajor caju (R. Naraian, 2002). In our investigation, it was found that beyond certain concentration, and biological efficiency was decreased and recorded to the minimum peptone 1 % in wheat straw (600 gram) Sivaprakasam (1980), however, stated that the increasing nitrogen level increased biomass production. This was probably due to competitors, appeared after first flush that exhausted nutrient material from the substrates. The sets supplemented with inorganic chemicals Mgso 4 was found that gave maximum and biological efficiency in 1.0% concentration in wheat

134 SHARMA Fig. 2 : Lactose 2% concentration in straw. Table 2 : Effect different concentration Nitrogen source (Peptone) on and Biological efficiency straw (880 grams and 88.0%). These results are similar to findings Verma (2005) who observed higher and biological efficiency in 0.5% concentration while working with Volvariella volvacea as a test fungus. He also stated that the use inorganic chemicals capable interacting with phenolics could protect the side chains extra cellular enzymes, important during the fruiting process as well as substrate utilization. The use micronutrient at low concentrations may intervene the importance in enzymatic reaction. Several workers also used inorganic sources in the supplementation various substrates, increasing Pleurotus spp. productivity. The sets treated with vitamin B- complex shows Peptone-1% 13 16 21 4 600 60.0 Peptone-2% 13 16 21 4 630 63.0 Peptone-3% 13 16 21 4 605 60.5 Peptone-1% 12 17 23 4 705 70.5 Peptone-2% 12 17 23 4 720 72.0 Peptone-3% 12 18 22 4 695 69.5 Peptone-1% 16 21 27 4 710 71.0 Peptone-2% 16 22 27 4 780 78.0 Grass Peptone-3% 16 22 29 4 900 90.0 Control 17 19 24 4 611 61.1 Table 3 : Effect different concentration Inorganic chemical (MgSo 4 ) on and Biological efficiency MgSo 4-1% 13 17 24 4 880 88.0 MgSo 4-2% 13 17 24 4 795 79.5 MgSo 4-3% 13 17 23 4 760 76.0 MgSo 4-1% 14 19 26 4 770 77.0 MgSo 4-2% 13 19 26 4 810 81.0 MgSo 4-3% 14 19 25 4 695 69.5 MgSo 4-1% 16 24 30 4 530 53.0 MgSo 4-2% 16 24 31 4 685 68.5 Grass MgSo4-3% 16 23 30 4 730 73.0 Control 15 20 27 4 688 68.8

EFFECT OF CHEMICAL SUPPLEMENTATION OF P. SAJOR CAJU 135 Table 4 : Effect different concentration Vitamin source (B-Complex) on and Biological efficiency Vit.B-100 mg/l Spray 12 14 19 4 765 76.5 Vit.B-200 mg/l Spray 12 14 19 4 780 78.0 Vit.B-300 mg/l Spray 12 15 19 4 860 86.0 Vit.B-100 mg/l Spray 13 16 22 4 692 69.2 Vit.B-200 mg/l Spray 13 17 21 4 696 69.6 Vit.B-300 mg/l Spray 14 18 22 4 705 70.5 Vit.B-100 mg/l Spray 16 20 26 4 760 76.0 Vit.B-200 mg/l Spray 16 19 26 4 765 76.5 Grass Vit.B-300 mg/l Spray 16 19 25 4 715 71.5 Control 15 17 23 4 690 69.0 Table 5 : Effect different concentration Growth promoter (EDTA) on and Biological efficiency EDTA-0.1% Spray 13 16 19 4 775 77.5 EDTA-0.2% Spray 13 16 19 4 900 90.0 EDTA-0.3% Spray 13 16 19 4 865 86.5 EDTA-0.1% Spray 12 17 21 4 690 69.0 EDTA-0.2% Spray 12 17 21 4 770 77.0 EDTA-0.3% Spray 12 17 21 4 685 68.5 EDTA-0.1% Spray 17 23 28 4 705 70.5 EDTA-0.2% Spray 17 24 28 4 716 71.6 Grass EDTA-0.3% Spray 17 24 30 4 690 69.0 Control 15 20 23 4 657 65.7 Table 6 : Comparative chemical analyses Pleurotus. sajor caju in different supplement and substrates (100 gm dry wt. basis) Contents - Lactose- 2% - Peptone-3% - MgSo 4-1% - EDTA-0.2% CONTROL - straw - straw - straw - straw Protein 47.96 47.90 47.94 47.99 47.93 Sugar 0.288 0.281 0.280 0.271 0.285 Starch 0.110 0.122 0.123 0.110 0.120 Fibers 1.077 0.984 1.109 1.092 1.084 Ash 0.990 0.991 0.969 0.973 0.974 Moisture 90.92 90.89 90.94 90.93 90.95

136 SHARMA maximum and biological efficiency in 300mg/ltr i.e. 860gram and 86.0% in wheat straw. The and biological efficiency increased with increasing order concentrations. All the sets took equal time for spawn and pin (13 days wheat straw, 12 days paddy straw and 17 days lemon grass), they showed lesser time for crop when treated with growth promoter EDTA (Table 5). Among the three lignocellulosic agricultural wasted used as substrate, best growth and high P. sajor caju was obtained in wheat straw. REFERENCES Bano, Z. and Rajarathnam, S. (1982). In tropical mushroom (eds. S.T. Chang and T.H.Quimio). The Chinese Univ. Press, Hong Kong, pp: 363-380. Sohi, H.S. (1992). Importance edible mushrooms in India diet and factors responsible for low production cultivated mushrooms, Indian phytopathology, 45(2): 147-157 (1992). Mehta, K.B. (1990). Mushroom Recipes, Technical Bulletin No.3 Solan, India, National Centre for Mushroom Research and Training. Pp-17-19. Munjal, R.L. (1982). Prospects mushroom cultivation in India. Indian Phytopathology, 35: 1-13. Singh Rochica (2005).Studies on some larger fungi Faizabad with reference to their eco-physiological characteristics. Ph.D. Thesis, Dr. R.M.L.Avadh Univ., Faizabad, pp- 114-115. Sivaprakasam, K. (1980). Studies on oyster mushroom (Pleurotus sajor caju (Fr.) Singer). P.h.D. Thesis, Tamil Nadu Agric. Univ. Coimbatore, pp- 115-116. Naraian, R. (2002). Effect nitrogen supplementation on the fruitbodies and changes in enzyme prile Pleurotus florida. Ph.D. Thesis, Dr. R M. L. Avadh University, Faizabad, pp-19-21. Upadhyay, R.C. and Fritsche, W. (1997). In Advance in Mushroom Biology and Production (eds. R.D. Rai, B.L. Dhar and R.N. Verma) MSI, Solan (HP), pp: 281-290. Verma, D.N. (2005). Studies on cultivation Volvariella volvacea (Bull. Ex. Fr.) Singer with special reference to certain organic and inorganic amendments, pp-23-27. Yadav, M.C., Dhar, B.L., Verma, R.N. and Kamal, S. (1998). International Symposium: Science and Cultivation Mushroom, Nanjing, October-1988, pp- 12-15.