Evaluation of Various Agro-Wastes for Production of Pleurotus spp. (P. florida, P. sajor-caju and P. eous)

P. florida, P. eous and P. sajor-caju was cultivated on different agro-wastes viz. paddy straw, wheat straw, sorghum straw, soybean straw, paddy + wheat straw (1:1), paddy + sorghum straw (1:1), paddy + soybean (1:1), wheat + soybean straw (1:1), wheat + soybean straw (1:1) and soybean + sorghum straw (1:1) to determine the effect of these agro waste on yield, growth and biological efficiency. Paddy straw showed significantly highest yield of 1248.3, 1348.7 and 1275.7 g/1.5 kg of dry substrate of P. florida, P. eous and P. sajor-caju respectively and biological efficiency of 83.22 %, 89.90 % and 84.99 % with minimum days for spwan run completion, pinhead initiation, development of mature fruiting bodies of P. florida (14, 15 and 15 days), P. eous (18, 19 and 19 days) and P. sajor-caju (21, 24 and 23 days) respectively.

Oyster mushroom (i.e.Pleurotus spp.) is commonly called as Dhingri in India because of its oyster like shape.Pleurotus is an efficient lignin degrading mushroom and can grow well on different types of lignocellulolosic materials.Different species of Pleurotus can grow well in variable temperature conditions; hence they are ideally suited for cultivation throughout the year in various regions of tropical country like India (Ahmed et al., 2009).For many reasons the fungi of the Pleurotus genus have been intensively studied in many different parts of the world; they have high gastronomic value.They are able to colonize and degrade a large variety of lignocellulosic residues, they require shorter growth time when compared to other edible mushrooms, they demand few environmental controls, their fruiting bodies are not very often attacked by diseases and pests and they can be cultivated in a simple and cheap way (Patrabansh and Madan, 1997).
An attractive feature of oyster mushrooms is that they can utilize a large variety of agricultural waste products and transform the lignocelluloses biomass in to high quality food, flavor and nutritive value (Quimio, 1978;Bano and Rajarathanam, 1982;Jain and Vyas, 2003).
Oyster mushroom posses the appropriate enzymatic mechanism for the transformation of complex organic macromolecules into simple compounds have been exploited as the means for biodegradation of a wide range of plant litter due to their particular ability for selective delignification (Mayson & Verachtert, 1991;Martinez et al., 1994).Most agricultural residues are rich in lignocelluloses compounds whose handling and disposal often problematic.Wheat straw, Soybean straw, Paddy straw and Sugarcane bagasses are the substrates of interest, Since they are produced in large quantities and rich in cellulose and lignin.The potential of bioconversion of lignocelluloses waste into value added products is emphasized in earlier studies (Philippoussis & Zervakis 2000; Poppe, 2000).
Mushrooms are consumed for their deliciousness and nutritive value.Mushrooms are excellent as a food as they provide a full protein diet containing twenty one amino acids besides vitamins and minerals.Being easily digestible (70-90%), mushroom protein is considered superior to vegetable proteins.Several mushrooms have been screened for their bio active properties and many compounds such as polysaccharides mainly ²-D glucan, heteroglycans, hexose correlated compounds, pachymanans, proteioglycans, terpenoides such as ganoderic acid, ganoderol, ganodermic acid and compounds like germanium and ergosterol have been isolated and commercialized (Hobbs, 1996;Wani et. al., 2010).
The commercial cultivation of various Pleurotus species including P. sajar-caju, P. eous, P. florida, P. flabellatus, P. ostreatus etc. Pleurotus florida is white oyster mushroom it is white in colour from primordial / pin head formation to maturity.The pileus of this mushroom with thin margin, smooth and pileus thickness is lesser as compares to P. ostreatus and P. sajor-caju.The mushroom looks like a white disc, growing on a thick stipe with decurrent gills extending to the base of the stipe.This mushroom grew excellently at 18-22 °C temperature range but can grow up to 28°C.
Various Pleurotus species have been shown to possess a number of medicinal properties, such as antitumor, immunomodulatory, antigenotoxic, antioxidant, anti-inflammatory, hypochloesterolaemic, antihypertensive, antiviral and antimicrobial activity (Gregori et al, 2007).

MATERIALS AND METHODS
Prepartion of pure culture of P. florida, P. sajorcaju and P. eous.
Matured pileus/cap of P. florida, P. sajorcaju and P. eous was placed in the sterile glass petriplats (90 mm) lined with dark black coloured drawing sheet paper, facing gills underside covered with lid and kept as such for a over night.Next day morning abundant white coloured circular spore print on paper sheet was obtained.From this spore prints, spores were gently lifted with the wire loop and transfered on autoclaved and cooled PDA medium in glass petriplates under Laminar air flow cabinet.These plates were then incubated at 20 0 C in an incubator.After a week of incubation, profused whitish, cottony growth was developed.From these plates, pure culture of P. florida, P. sajor-caju and P. eous were preapred on PDA slants in glass test tubes and preserved in refrigrator.

Prepartion of master spawn
Aperantly healthy, unbroken and clean wheat/bajra grains were paraboiled in clean water (1:1 w/v).After boiling, excess water was drained of by spreading the grains on wire mesh/sieve.Then these were spread on surface steralized (4% formalin) polythene sheet to which calcium sulphate @ 2% and calcium carbonate @ 5% were mixed on dry weight basis.These grains were filled in glass conical flasks (200 g/flask), plugged with non-absorbant cotton and then steralized in autoclave at 15 Lbs pressure for 20 min, for two consecutive days.After steralization, the flasks were transfered to Laminar-Air-Flow Cabinet, allowed to cool at room temperature and inoculate with 4-5 mycelial discs (5mm) of pure cultures of P. florida, P. sajor-caju and P. eous and incubated at 20 0 C.These flasks were shaken intermittently to faciliate through spreading on the mushroom mycelium on the grains.After three weeks of incubation, the grains in flasks were fully covered with the mycelium of P. florida, P. sajor-caju and P. eous.Thus the master spawn was prepared.

Prepartion of commercial spawn
Commercial spawn was prepared using 250-300 guage polypropylene bags 8 x 12 inch 2 wheat/bajra grains are paraboiled, amended with Calcium sulphate and Calcium carbonate as detailed under 3:3:2 filled in polypropylene bags (200 g/bag), closed the openings of bags using PVC pipe (2.5cm dia.) piece and plugged the pipes with non absorbant cotton and autoclaved at 15 Lbs pressure for 20 minutes.The autoclaving was done for two consecutive days.Later these bags were shifted to Laminar-Air-Flow Cabinet, allowed cool at room temperature and inoculted with a sponful of master spawn and incubated with at 20°C temperature.The bags were shaken intermittently to facilate through spread of mushroom mycelium.During incubation of the bags were regularly examined for mould infestation if any and contaminated bags were immadiately discarded to avoid build up of contamination in the vicinity.Within 18-20 days of incubation, mushroom mycelium was fully covered on the grains surface and thus the commercial spawns of P. florida, P. sajor-caju and P. eous were preapred .These commercial spawn bags were stored in refrigrator and used for spawning the substrates.

Evaluation of agro-wastes
Standard polythene bag method of Pleurotus spp.mushroom cultivation (Zakia et al. 1979).

Substrate preparation Physical sterilization of straws
The following substrates are used to cultivation i.e. paddy straw, wheat straw, sorghum straw, soybean straw, paddy + wheat straw (1:1), paddy + sorghum straw (1:1), paddy + soybean (1:1), wheat + soybean straw (1:1), wheat + soybean straw (1:1) and soybean + sorghum straw (1:1).The dry straws substrates were chopped to small pieces (3-5 cm long).The chopped substrate were weighed and then soaked in cold water for 12 hours.After soaking substrates were be taken out and excess of water drained off.After draining excess of water these straws were weighed.These straws were then sterilized in autoclave at 20 Ibs psi for 20 minutes.After autoclaving the straws were cooled down to ambient temperature and used for filling the polythene bags.

Cultivation steps
The polypropylene bags of the size 35 × 55 cm 2 (100 gauge thickness) will be sterilized by dipping in 2 % formalin prior to use and lower corners of the bags was tied with the string so that the bed assumes a round shape after filling the straw and were filled with sterilized substrates and multilayered spawning @ 2 percent of wet weight of the substrate.The bags was filled up to their 90 percent capacity and mouths will be closed tightly with threads with the help of sterilized needle, about 20-25 minute holes all round the filled bags was made.
A spawned substrate bag was kept in mushroom house where the temperature and humidity were maintained around 20-25ÚC and 80-90 %, respectively with sufficient light and ventilation for 20 days.After completion of spawn run the bags were removed by cutting longitudinally with sharp blade and these beds was kept on bamboo racks/platform at 15-18ÚC temperature and 80-90 % relative humidity for cropping.Pinhead initiation was evident within 3-4 days after removal of poly bags.The beds were maintained up to the harvest of the third flush, which was completed in 35-40 days after sowing.A small layer of substrate was scrapped off from all the side of the beds after each harvest.

RESULTS AND DISCUSSION
Evaluation of various agro-wastes for production of P. florida, P. eous and P. sajor caju.The experiment on productivity were carried out to study the effect of different substrates i.e. paddy straw, wheat straw, sorghum straw, soybean straw, paddy + wheat straw (1:1), paddy + sorghum straw (1:1), paddy + soybean (1:1), wheat + soybean straw (1:1), wheat + soybean straw (1:1) and soybean + sorghum straw (1:1) on number of day required for completion of spawn run, number of days required for pinhead intiation, number of days required for development of matured fruiting bodied, number of pinhead, number of matured fruiting bodies, diameter of stipe, pileus diameter, yield, moisture content, dry weight and biological efficiency of P. florida, P. eous and P. sajor-caju.

Days required for completion spawn run, pinhead initiation and matured fruiting bodies
The data on average number of days required for completion of spawn run, pinhead initiation and matured fruiting bodies of P. florida, P. eous and P. sajor-caju in the table 1, 2 nad 3. The analyzed of variance indicated that the treatments significantly affected the number of days required for completion of spwan run of P. florida, P. eous and P. sajor-caju .It was observed that treatment T 1 (100 % Paddy straw) required minimum days (14, 15 and15 days) to complete spawn run followed    The number of days required for pinhead initiation of P. florida, P. eous and P. sajor caju on different substrates were recorded.Paddy straw took minimum number of days in tratment T 1 (18,19 nad 18 days) to produce pinhead in mushroomrespectively.While highest number of days were taken in T 7 (50 % paddy straw + 50 % sorghum straw; 25 days) for P. florida , T 10 (50 % soybean straw + 50 % sorghum straw; 30 days) for P. eous and T 8 (50 % wheat straw + 50 % soybean straw; 28 days) for P. sajor caju respectivly.
The number of days required for matured fruiting bodies of P. florida on different substrates were recorded.Minimum number of days required for development of mature fruiting bodies was 21, 24 and 23 days was recorded in the treatment T 1 in which paddy straw was used respectively.The maximum number (28 days) of days to reach maturity stage were observed in T 5 (50 % paddy straw + 50 % sorghum straw; 28 days) for P. florida, T 10 (50 % soybean straw + 50 % sorghum straw; 35 days) for P. eous and T 8 (50 % wheat straw + 50 % soybean straw; 32 days) P. sajor caju respectivly.
Similar variation in days required for spawn run, pinhead initiation and matured fruiting bodies has been recorded by earlier workers (Iqbal et. al, 2011;Jafarpour et. al, 2011;Musieba et. al, 2012;and Pala et. al, 2012).

Average number of pinhead/ bed and matured fruiting bodies/bed
The average number of pinhead and matured fruiting bodies of P. florida, P. eous and P. sajor-caju on different substrates were recorded and showed in table 4. The maximum average number of pinhead 140.33 mm, 142.67 mm and 147.33 mm were recorded in the treatment T 1 in which Paddy straw was used as a substrate for P. florida, P. eous and P. sajor-caju respectivly and The minimum number of pinhead were recorded in T 8 (wheat straw + soybean straw; 117.33 mm), T 9 (wheat straw + sorghum straw; 121.33 mm and T 10 (soybean straw + sorghum straw; 121.67 mm) for P. florida, P. eous and P. sajor-caju respectivly.
The greater number of pinhead is due to moisture content present in the growing substrates.Similar variation in number of pinhead has been reported by several workers in the past (Mondal et. al, 2010;Patil et. al, 2012).
The average number number of matured fruiting bodies/bed of P. florida, P. eous and P. sajor-caju were recorded the maximum matured fruiting bodies (116.33 mm, 121.33 mm and 118.67 mm) were obtained in treatment in which paddy straw was used as a substrates respectivly.The minimum average matured fruiting bodies were recorded in the treatment in which T 9 wheat straw + sorghum straw (95.33 mm), T 10 soybean straw + sorghum straw (92.66 mm) and T 10 soybean straw + sorghum straw (92.33 mm) was used as a substrate respectivly.
The maximum number of fruiting bodies was recorded on Paddy straw amongest the different substrates in P.florida, P.eous and P.sajorcaju.The least number of matured fruiting bodies was recorded with wheat + sorghum straw in P.florida and soybean + sorghum straw in P. eous and P. sajor-caju.Similar variation in number of matured fruiting bodies has been recorded by several workers (Jadhav et. al, 1996;Patil et. al, 1996;Dhoke, 1998;Mandhare, 2000, Mondal et. al, 2010and Survase, 2012).

Average pileus diameter and stipe diameter
The data pertaining to pileus diameter of P. florida, P. eous and P. sajor-caju on different substrates was presented in Table 3.
The average pileus diameter of P. florida, P. eous and P. sajor-caju on different substrates was recorded The highest average pileus diameter (9.33 cm², 12 cm² and 12 cm² ) was obtained in the treatment in which paddy straw used for three spp respectively.The lowest average diameter of pileus was recorded in the treatment T 7 paddy straw + sorghum straw (3.66 cm²), T9 wheat straw + sorghum straw (2.33 cm²) and T8 wheat straw + soybean straw (4 cm²) for P. florida, P. eous and P. sajor-caju on different substrates respectively.
The average diameter of stipe of P. florida, P. eous and P. sajor-caju on different substrates was recorded that the data clearly indicated that significantly highest average stipe diameter (3.66 cm², 4 cm 2 and 4.33 cm 2 ) in treatment in which paddy straw used as a substrate respectivly and the lowest average stipe diameter was recorded in treatment T9 wheat + sorghum straw, T8 wheat + soybean straw (1.66 cm 2 ) and T7 paddy + sorghum straw (1.33 cm 2 ) used as substrate respectively for the following three spp P.florida, P.eous and P.sajor-caju.
The pileus diameter and stipe diameter of P.florida, P.eous and P.sajor-caju has been influenced by different substrates and here to the similar observation were recorded.The pileus diameter and stipe diameter of P.florida, P. eous and P. sajor-caju maximum on Paddy straw amongst the different substrates and least on wheat + sorghum straw and soybean + sorghum straw.Such a variation in size of pileus and stipe of P.florida, P.eous and P.sajor-caju due to different substrates was recorded by different workers in the past (Patil, 1996;Dhoke, 1998;Mandhare, 2000;Mane et. al, 2007;Mondal et. al, 2010 andPatil et. al, 2012).

Fresh yield of mushroom
The average yield of P.florida, P.eous and P.sajor-caju on different substrates was recorded as follow.The mean data revealed that the maximum yield (1248.33 g/1.5 kg dry substrate) for P.florida ,(1348.33g/1.5 kg dry substrate) for P.eous and (1275 g/1.5 kg dry substrate) for P.sajor-caju was recorded in treatment T 1 in which paddy straw used as a substrate respectivly, which was found to be higher than all other treatment and the minimum total yield (795 g/1.5 kg dry substrate) was weighed in treatment T8 wheat straw + soybean straw used as substrate for P.florida, (856 g/1.5 kg dry substrate) in T8 wheat straw + soybean straw for P.eous and (759 g/1.5 kg dry substrate) was weighed in treatment T10 in which soybean straw + sorghum straw used as substrate for P.sajor-caju respectivly.
The yield of P. florida, P. eous and P. sajor-caju has been influenced by different substrates.The yield of P. florida was maximum with paddy straw and followed by Paddy + wheat straw and Wheat straw and least on Wheat + soybean straw.The yield of P. eous was maximum with paddy straw and followed by Wheat straw and Paddy + wheat straw and least on Wheat + soybean straw.The yield of P. sajor-caju was maximum with paddy straw and followed by Wheat straw and Paddy + wheat straw and least on Wheat + sorghum straw.Similar diffrential yield of P. florida, P. eous and P. sajor-caju with different substrates has been reported by ( Kirbag and Akyuz., 2008;Pandey et. al, 2008;Ingale and Ramteke., 2010;Jafarpour et. al, 2011;Raja andGanesh, 2013 andSharma et. al, 2013).

Dry weight of mushroom
The average dry weight of P. florida, on different substrates was recorded as follow.The maximum average dry weight (125.22 g) was noticed in treatment T5 in which paddy straw + wheat straw used as a substrate Minimum dry weight (89.33 g) was recorded in treatment T8 in which wheat straw + soybean straw used as a substrate.The average dry weight of P. eous on different substrates was recorded in between 80.33 to 139.22 g.The maximum average dry weight (139.22 g) was noticed in treatment T2 in which wheat straw used as a substrate and Minimum dry weight (80.33 g) was recorded in treatment T10 in which soybean straw + sorghum straw used as a substrate and average dry weight P. sajor-caju on different substrates the maximum average dry weight (122.66 g) was noticed in treatment T1 in paddy straw used as a substrate and Minimum dry weight (80.55 g) was recorded in treatment in which wheat straw + soybean straw used as a substrate.
The average dry weight of P.florida, P.eous and P.sajor-caju differed with substrates used for cultivation.The highest dyr weight of P. florida was recorded with Paddy + Wheat straw and followed by Wheat straw amongst different substrates.The average dry weight of P. florida was recorded in present investigatin ranged between 125.22 -91.06 g.The highest dyr weight of P. eous was recorded with Wheat straw amongst different substrates.The average dry weight of P. eous was recorded in present investigatin ranged between 139.22 -80.33 g.The highest dyr weight of P. sajor-caju was recorded with Wheat straw amongst different substrates.The average dry weight of P. sajor-caju was recorded in present investigatin ranged between 120.3 -80.55 g. similar variation in respect of dry weight of P.florida, P.eous and P.sajor-caju were reported by different workers in the past ( Ingale and Ramteme, 2010 and Patil et. al, 2012).

Moisture content of mushroom
The moisture content of dehydrated air dried mushroom of P. florida, P. eous and P. sajorcaju had been estimated and are presented in Table 16.
The result indicated that the moisture In present investigation, highest moisture content of P.florida was evident with Wheat + Soybean straw amongst the substrates.In P. eous highest moisture content was evident with paddy straw and in P. sajor-caju highest moisture content was evident with paddy straw followed by wheat + soybean straw.Similar variation in moisture content of Pleurotus species has been reported in the past (Dundae et. al, 2009;Sayed et. al, 2009;Hassan et. al, 2010 andPatil et. al, 2010).

Biological efficiency of P. florida, P. eous and P. sajor-caju
The effect of substrates on yield contributing charecters such as biological efficiency was varied ( Table 5).
It was necessary to calculate percentage of biological efficiency because certain substrates were denser than other.The conversion percentage from dry substrate weight to fresh mushroom weight (biological efficiency) was determined.The different substrates showed different responde on biological efficiency of mushroom (Table 16).
In P. florida, P. eous and P. sajor-caju maximum biological efficiency 83.22 %, 89.90 % and 84.99 % of mushroom was noticed in treatment in which paddy straw used as substrate respectively and minimum biological efficiency (52.99 %) of mushroom was recorded in treatment T8 wheat straw + soybean straw used as substrate in P. florida, (57.06 %) in treatment T8 wheat straw + soybean straw used as substrate in P. eous and (50.59 %) of mushroom was recorded in treatment T10 soybean straw + sorghum straw used as substrate in P. sajor-caju respectivly.
Similar diffrential biological efficiency of P. florida, P. eous and P. sajor-caju with different substrates has been reported by (

CONCLUSION
The result of present investigation revealed that in P. florida, P. eous and P. sajor-caju the number of days required for spawn run, pinhead initiation and matured fruiting bodies was minimum on paddy straw.The number of pinhead and matured bodies was maximum on Paddy straw.Stipe and pileus are important edible parts of mushroom.Highest pileus diameter and stipe diameter was recorded on paddy straw.The average yield and biological efficiency of matured fruiting bodies was maximum on Paddy straw amongst the different substrates.Maximum dry weight was recorded on Paddy + Wheat straw in P. florida and P. sajor-caju and wheat straw in P. eous.The highest moisture content was recorded on wheat + soybean straw in P. florida and paddy straw in P. eous and P. sajor-caju.Paddy straw for cultivation of P. florida, P. eous and P. sajor-caju superior

Table 1 .
Effect of various substrates on vegetative and reproductive growth of P. florida, P. eous and P. sajor-caju

Table 2 .
Effect of various substrates on number of pinhead, matured fruiting bodies, size of pileus diameter and stipe diameter of P. florida, P. eous and P. sajor-caju