Monika Final Thesis 6th September 2024 PDF

Evaluation of biological efficiency of Pleurotus spp. on different agro-wastes in Haryana By Monika (2022A83M) Thesis submitted to Chaudhary Charan Singh Haryana Agricultural University, Hisar in partial fulfil...

Evaluation of biological efficiency of Pleurotus spp. on different agro-wastes in Haryana By Monika (2022A83M) Thesis submitted to Chaudhary Charan Singh Haryana Agricultural University, Hisar in partial fulfillment of the requirements for the degree of MASTER OF SCIENCE IN PLANT PATHOLOGY DEPARTMENT OF PLANT PATHOLOGY COLLEGE OF AGRICULTURE CCS HARYANA AGRICULTURAL UNIVERSITY HISAR– 125004, HARYANA 2024  CERTIFICATE – I This is to certify that this thesis entitled “Evaluation of biological efficiency of Pleurotus spp. on different agro-wastes in Haryana” submitted for the degree of Master of Science in the subject of Plant Pathology to the Chaudhary Charan Singh Haryana Agricultural University, Hisar is a bonafide research work carried out by Ms. Monika, Admission No. 2022A83M, under my supervision and that no part of this thesis has been submitted for any other degree. All the assistance and help received during the course of investigation have been fully acknowledged. Dr. Satish Kumar (Major Advisor) Joint Director (Plant Pathology) SNIATTE CCS Haryana Agricultural University Hisar-125004 (Haryana)  CERTIFICATE-II This is to certify that this thesis entitled “Evaluation of biological efficiency of Pleurotus spp. on different agro-wastes in Haryana” submitted by Ms. Monika, Admission No. 2022A83M to the Chaudhary Charan Singh Haryana Agricultural University, Hisar in partial fulfillment of the requirement for the degree of Master of Science in the subject of Plant Pathology has been approved by the Student‘s Advisory Committee after an oral examination on the same. MAJOR ADVISOR EXTERNAL EXAMINER HEAD OF THE DEPARTMENT DEAN, POSTGRADUATE STUDIES  CERTIFICATE–III FORMAT FOR P.G.THESIS “It is certified that the thesis submitted by Ms. Monika, Admission No. 2022A83M, M.Sc. student of Plant Pathology department has been checked and found as per specification of the format circulated by the Dean, PGS vide Memo No. PGS/A1/09/6926-90 dated 26.8.2009”. MAJOR ADVISOR HEAD OF THE DEPARTMENT  Acknowledgements I am profoundly thankful to the divine, the merciful, for bestowing upon me the fortitude and bravery required to finalize this manuscript. I am immensely pleased to convey my profound respect and gratitude to my major advisor, Dr. Satish Kumar, Joint Director (Dept. of Plant Pathology), SNIATTE. His motivating guidance, generous assistance, steadfast encouragement, and thoughtful recommendations were crucial in reviewing the manuscript and providing essential feedback. His support played a key role in enabling me to complete this project on schedule. I am truly grateful and offer my sincerest thanks to him. I extend my deepest appreciation to member of my co-major advisor, Dr. Jagdeep Singh (Assistant Scientist, Dept. of Plant Pathology), for their noble guidance and helping nature. I also extend my gratitude to my esteemed Advisory Committee members Dr. Krishna Rolania (Dean PGS’ nominee), Assistant Scientist, Dept. of Entomology; Dr. Sardul Mann, Assistant Professor, Dept. of Nematology; Dr. Manoj Kumar, Assistant Scientist, Dept. of Maths and Statistics, for their invaluable insights, thorough evaluation and unwavering support throughout the course of this investigation. I sincerely extend my deep gratitude to Dr. Anil Kumar (Professor and Head of the Dept. of Plant Pathology), for providing the essential facilities to conduct this study. I also wish to express my deep appreciation to the faculty and non-teaching staff of the Dept. of Plant Pathology for their unwavering support throughout this research. Additionally, I am grateful to Mr. Mandeep, Laboratory Assistant at the Mushroom Technology Laboratory, CCS Haryana Agricultural University, Hisar, for his generous support at all times. I would like to express my affection to my seniors Dr. Chaman, Dr. Deepak Kaushik, Mr. Mohit, Mr. Amit Bhardwaj, Mr. Ashok Sharma, Mr. Rohit, Mrs. Simran Jeet, Mr. Sourabh for your kindness, guidance and encouragement, as well as for your timely assistance whenever I needed it. I want to extend my deepest thanks to all my dear friends Deepak, Ravinder, Pooja, Komal and lovely juniors Jyoti, Renu, Pooja for their timely help, support, a great company and for making my college life truly unforgettable. I want to express my deep sense of gratitude to my father Sh. Amar Singh, mother Smt. Krishna Devi, brothers Mr. Ashvani and Dr. Pradeep & sister-in-law Mrs. Pooja for their everlasting love, endless support, selfless care, moral inspiration and blessings have consistently create confidence in me to complete this work. I also want to express my heartfelt love for my nephew Mr. Rakshit Sharma. I just wanted to let you know how much happiness you bring into my life. Your giggles and mischievous activities make every day extra special. You’re my little star, and I’m lucky to be your Bhua! I’d like to express my heartfelt gratitude to my doggie Situ whose unconditional love and joyful spirit have brought so much happiness into my life. Your playful antics and gentle companionship are deeply appreciated. Thank you for being such a such a treasured and loving part of my life. My sincere thanks also goes to CCS Haryana Agricultural University, Hisar for providing me an opportunity of higher studies, which will be highly helpful in my future career. Date: July, 2024 Place: Hisar Monika  CONTENTS CHAPTERS DESCRIPTION PAGE (S) 1. INTRODUCTION 1-3 2. REVIEW OF LITERATURE 4-11 3. MATERIALS AND METHODS 12-19 4. RESULTS 20-46 5. DISCUSSION 47-56 6. SUMMARY AND CONCLUSION 57-61 7. BIBLIOGRAPHY i-vii  LIST OF TABLES Table Title Page(s) No. Effect of chemically pasteurized substrates and their 4.1 22 supplementation on crop cycle of Pleurotus djamor Effect of chemical pasteurised substrates and their supplementation 4.2 24 on growth characteristics and yield of P. djamor Effect of heat pasteurised substrates and their supplementation 4.3 26 on crop cycle of P. djamor Effect of heat pasteurised substrates and their supplementation on 4.4 28 growth characteristics and yield of P. djamor Effect of chemical pasteurised substrates and their supplementation 4.5 31 on crop cycle of P. pulmonarius Effect of chemical pasteurised substrates and their supplementation 4.6 33 on growth characteristics and yield of P. pulmonarius Effect of heat pasteurised substrates and their supplementation on 4.7 35 crop cycle of P. pulmonarius Effect of heat pasteurised substrates and their supplementation on 4.8 37 growth characteristics and yield of P. pulmonarius Evaluation of BC ratio upon cultivation of P. djamor on chemical 4.9 41 pasteurised substrates and their enrichment with substrates Evaluation of BC ratio upon cultivation of P. djamor on heat 4.10 42 pasteurised substrates and their enrichment with substrates Evaluation of BC ratio upon cultivation of P. pulmonarius on 4.11 44 chemical pasteurised substrates and their enrichment with substrates Evaluation of BC ratio upon cultivation of P. pulmonarius on heat 4.12 45 pasteurised substrates and their enrichment with substrates  LIST OF FIGURES Figure DESCRIPTION Page(s) No. Effect of substrates alongwith supplements and pasteurized by 4.1 different methods on crop duration and yield of Pleurotus 29 djamor Effect of substrates alongwith supplements and pasteurized by 4.2 different methods on crop duration and yield of Pleurotus 38 pulmonarius Estimation of BC ratio and Net returns of cultivation of 4.3 Pleurotus djamor on different supplements and pasteurised by 43 different methods Estimation of BC ratio and Net returns of cultivation of 4.4 Pleurotus pulmonariuis on different supplements and pasteurised 46 by different methods  LIST OF PLATES Plate DESCRIPTION Page(s) No. 1. (a-g) Substrate prepration and spawning 17 2. (a-d) Fruiting in P. djamor 27 3. (a-d) Fruiting in P. pulmonarius 36 LIST OF ABBREVIATIONS % : Per cent & : And @ : At the rate of / : Per et. al. : (et ali or et alia) and others g : Gram kg : Kilogram h : Hours min : Minutes pH : Potenz of Hydrogen viz. : videre licet e.g. : Exempli gratia i.e. : id est Fig : Figure ppm : Parts per million WP : Wettable Powder °C : Degree Celsius psi : Per square inch RBD : Randomized Block Design PDA : Potato Dextrose Agar ₹ : Rupees (Indian currency) CHAPTER-I INTRODUCTION Mushrooms, classified as Basidiomycetes, are the saprophytic fungi that thrive in damp environment and decompose organic material. Their significance lies in their crucial role in nutrient cycling (Subramanian, 1995). These are macro fungi with visible fruiting bodies, falling into four main categories: edible, medicinal, poisonous and a miscellaneous group with less defined properties, tentatively labelled as "other mushrooms" (Chang and Miles, 2004). Mushrooms are recognised as a delicious and nutritionally valuable food due to their high contents (per 100 g) of protein (23.22 g), carbohydrates (63.17 g), fibre (34.0 g), lipid content (4.71 g), total solids (9.37 g) on dry weight basis and vitamin C and phosphorus (6.67 and 107.13 mg per 100 g) on wet weight basis (Furlani and Godoy, 2007). The fresh mushrooms contain 85-95 per cent moisture, 35-70 per cent carbohydrates, 15-34.70 per cent protein, 6-10.90 per cent minerals, 10.00 per cent fat, 3-8 per cent nucleic acid on dry weight basis (Assemie and Abaya, 2022). In addition, mushrooms have also been found to have terpenoids, phenolic compounds, steroids, lectins, vitamins, antioxidants like ergothrionine and glutathione etc. In addition, it contains zinc, selenium, magnesium, calcium, manganese, iron, etc. and bioactive compounds like polysaccharides, carbohydrate-binding proteins, peptides, proteins, enzymes, polyphenols, triterpenes and triterpenoids (Alam et al., 2007, Seo and Choi, 2021). Commonly referred to as oyster mushrooms, Pleurotus species are edible fungi that are grown all over the world, primarily in South East Asia, Europe, Africa and India. The word Pleurotus comes from the Greek word pleuro, which means "laterally formed" or "lateral position of the stalk or stem." Pleurotus is a member of the class Agaricomycetes, family Pleurotaceae of the order Agaricales, subphylum Agaricomycotina of the Phylum Basidiomycota and subkingdom Dikarya of the Kingdom Fungi (Kirk et al., 2008). A few species of Pleurotus are P. ostreatus, P. columbinus, P. florida, P. salignus, P. spodoleucus and P. pulmonarius; the subspecies are P. sajor-caju, P. sapidus, P. populinus, P. calyptratus, P. dryinus, P. cornucopiae, P. djamor, P. flabellatus, P. eryngii, P. cystidiosus and P. tuber- regiu (Nadir et al., 2016). The total mushroom production in the world is 48.33 million tonnes during 2022 (FAOSTAT, 2022) with Lentinula edodes (shiitake) contributing 26.00 per cent, Auricularia spp. 21.00 per cent, Pleurotus ostreatus 16.00 per cent, Agaricus bisporus 11.00 per cent. India is the sixth largest producer of mushroom in the world (Singh et al., 2020). The oyster mushrooms contribute about 17 per cent of total mushroom production in India (Sharma et al., 2017). According to first advance estimates of 2023-24, it was found that total mushroom 1 production was 333680 metric tonnes in India whereas in Haryana it was 23900 metric tonnes (Anonymous, 2023). As per data available at Ministry of Agriculture and Farmers Welfare, Govt. of India during 2023-24, Bihar is the leading state in total mushroom production (39010 metric tonnes) followed by Odisha (34600 metric tonnes) and Maharashtra (30770 metric tonnes). Pleurotus spp. can be easily artificially cultivated and is endemic to temperate and tropical forests where it grows on dead and decomposing organic detritus, wooden logs and occasionally on dying deciduous or coniferous tree trunks (Randive, 2012). The commercial farming of mushrooms globally experienced a surge only after the conclusion of World War II (Singh et al., 2017). Pleurotus species are grown extensively on a large scale, utilizing a broad spectrum of agricultural substances, employing uncomplicated and cost-effective production techniques (Jegadeesh et al., 2018). These mushrooms can be easily cultivated on various agro-wastes like wheat straw, mustard straw, paddy straw, saw dust, cotton wastes etc. but their biological efficiency and other growth characteristics of different Pleurotus species vary according to substrates used with supplements. India produces about 500 million tonnes of crop residue per year as per Ministry of New and Renewable Energy (MNRE), Government of India, New Delhi (Anand and Kaur, 2024) The bulk of crop residue is consumed as feed, fuel and other industrial uses, but still there is a surplus of 140 million tonnes and out of these 92 million tonnes is burnt (Meena et al. 2022). The Pleurotus species demonstrated its adaptability for cultivation in both small- and large-scale farms across tropical and temperate climates, utilizing diverse lignocellulosic agricultural and forest wastes as substrates (Chang and Miles, 2004). Agro-industrial wastes possess the potential to be utilized in mushroom cultivation, thereby aiding in the alleviation of the malnutrition issues and potentially mitigating the environmental pollution (Ferdousi et al., 2020). The impact of substrate additives on the growth of oyster mushrooms can result in more productive and economical farming techniques (Sharma et al., 2023). The addition of rice bran, dry azolla, neem cake, dry biogas slurry and vermi-wash to rice straw has a significant positive effect on the oyster mushrooms' crude protein, total free amino acid, total carbohydrate and vital mineral nutrients including N, P and K (Elizabeth and Paul, 2020). Substrates require chemical and biological supplements since they only contain a portion of the nutrients that they need. The production of mushrooms can be made more productive by adding nutrient supplements to the substrates used in the process (Dubey et al., 2019). The cultivation of oyster mushrooms is characterized by its minimal spatial requirements. Therefore, it is regarded as one of the most productive sectors with a high benefit-cost ratio. Gaining more expertise in value-adding is also recommended to boost 2 producer profits (Quion et al., 2023). Economic profitability research revealed that growing Pleurotus mushrooms was a viable source of revenue for farmers and unemployed youth (Singh et al., 2018). The problem of burning of crop residues of wheat, paddy, sugarcane, mustard etc. is very common in Haryana, Punjab, U.P. etc. and therefore, it needed conversion to reusable resources like mushroom production for environment and to achieve nutritional security. Thus, keeping in mind the use of different agro-wastes to optimize the yield of different oyster mushrooms in Haryana, the present study was carried out with the following objectives: 1. To study the growth characteristics and yield of Pleurotus djamor and Pleurotus pulmonarius on different agro-wastes. 2. To evaluate the benefit-cost ratio of P. djamor and P. pulmonarius on different agro- wastes. 3 CHAPTER-II REVIEW OF LITERATURE Knowledge of mushrooms dates back to the "VEDA" era. Later, the fleshy-capped fungus in India were referred to as Chattri (Payak, 1969). Oyster mushrooms are renowned as one of the most extensively farmed edible fungi worldwide, attributed to their nutritional advantages and capability to flourish on a diverse array of agricultural waste (Sanli and Peksena, 2020). The basidiocarps of oyster mushrooms are characterized by their shell-like or oyster-shaped appearance, displaying a variety of colours such as white, cream, grey, yellow, pink or light brown. Pleurotus, a member of the phylum Basidiomycota, is farmed extensively worldwide because of its enhanced biological efficiency and inexpensive production method (Mane et al., 2007). Pleurotus species grown on non-compost and compost substrates showed varying degrees of biological effectiveness i.e. 45.33 % –120.07 % (Jatwa, 2016). The Pleurotus genus has the most farmed species of mushrooms due to its wide adaptability to temperature and climatic conditions (Zadrazil and Dube, 1992). Pleurotus djamor favours tropical and subtropical areas and is recognized for its quick fruiting, capacity to thrive on various substrates and ability to withstand high temperatures. It demonstrates exceptional adaptability within the Basidiomycota phylum by thriving on a diverse array of forest and agricultural waste material (Stamets, 2000). Pleurotus pulmonarius exhibits a broad distribution across temperate and sub-tropical forests globally. This warm-weather variant is primarily cultivated in Europe and North America (Zhang et al., 2005). Since limited work has been done on evaluation of biological efficiency of Pleurotus spp. through different agro-wastes in India as well as in other countries, therefore the review of literature pertaining to it has also been reviewed under the following headings: 2.1 Effect of substrates 2.2 Effect of supplements 2.3 Effect of substrate pasteurisation 2.4 Yield and yield attributing characters 2.5 Benefit:Cost Ratio 2.1 Effect of substrates Champawat and Chitale (2003) in his research study found that maximum yield was seen for P. citrinopileatus grown on mustard straw, followed by sorghum and bajra straws. Shah et al. (2004) compared yield attributing characters of oyster mushroom (P. ostreatus) on different substrates and found that fruiting body of oyster mushroom differed with respect to stipe length and girth and pileus width when grown in different farm substrates. Adebayo et al. (2009) assessed the productivity of P. pulmonarius across various combinations of cotton waste and cassava peel, concluding that cotton waste exhibited superior performance for cultivating it as compared to cassava peel. Chaubey et al. (2010) documented that among the 4 traditional substrates, wheat straw exhibited significant aptness, showcasing a biological efficiency of 72.0 per cent for the cultivation of P. djamor. Dehariya et al. (2010) assessed various mixtures of agricultural and household refuse as potential substrates. Their findings indicated that soybean straw was optimal among traditional substrates, while domestic waste was effective among unconventional substrates. They revealed that combinations of soybean straw + wheat straw, soybean straw + domestic waste and domestic waste + used tea leaves had resulted in increased mushroom yield. Kumar et al. (2012) examined three distinct substrates, namely wheat straw (WS), maize cob straw (MCS) and lentil straw (LS) either individually or in combination with WS in ratios of 1:2, 1:1 and 2:1 alongside nine supplements including wheat bran, rice bran, corn meal, soybean meal, groundnut cake, pigeon pea powder, lentil powder, mustard cake and neem cake, and assessed the yield performance as well as biological efficiency of P. sajor-caju, P. florida, P. flabellatus and P. sapidus. Among all the tested Pleurotus spp., the substrate combination of WS+LS (2:1) exhibited the shortest spawn run time and produced the highest yield of P. sajor-caju (572.80g with 57.30% BE) followed by P. florida (554.20g with 55.40% BE). Among all the individual and combined ratio of substrate, the WS+LS (2:1) was found to be superior. Dubey et al. (2019) conducted a study focusing on Pleurotus mushrooms to assess the influence of different substrates on the performance of P. sajor-caju. The investigation involved the utilization of rice straw (T1), wheat straw (T2), banana leaves (T3) and sugarcane bagasse (T4) as treatment options for mushroom cultivation. The duration required for colonization, spawning 19 days, exhibited notable reductions with wheat straw and banana leaves, while the fruiting phase of 20.5 days was notably observed when wheat straw was employed. The elongation of the stipe at 4.9 cm was most pronounced with rice straw succeeded by banana leaves, wheat straw and sugarcane bagasse at 4.3 cm, 3.7 cm and 3.3 cm, respectively. Similarly, the largest cap diameter was documented with rice straw at 5.1 cm, followed by wheat straw, banana leaves and sugarcane bagasse, measuring 4.1 cm, 3.5 cm and 3.3 cm, respectively, under standardized environmental conditions and cultural methodologies across all substrates. Siwulski et al. (2019) in their study on cultivation of C. maxima, G. lucidum and L. edodes resulted in higher yield and mineral content when substrate based on alder and beech sawdust was used, cultivation of A. cylindracea and F. velutipes yielded better results on substrate based on oak sawdust and flax shives, P. eryngii cultivation outcomes were similar for both substrates. The study indicated that the chemical composition of substrates could affect both the yield and the level of various toxic and nutritional elements. Alam et al. (2020) in their research on effects of composition, age and sterilization techniques of mother culture on the growth and yield of Volvariella volvacea Singer reported that the highest biological efficiency (BE) for V. volvacea was achieved using a blend of rice straw and wheat bran as the substrate. Agba et al. (2021) assessed the effects of dry plantation leaves, sawdust as well as dry plantation leaves along with sawdust in 1:3, 2:3, 3:2, 1:1 and 4:1 ratio on various 5 growth parameters, total yield and biological efficiency of P. ostreatus. They noticed that sawdust substrate provided the optimum mycelia running rate whereas larger stipes were seen on dried plantation leaves. They concluded that sawdust substrate outperformed other substrates in yield, total fruiting bodies and biological efficiency. 2.2 Effect of supplements Sangeetha (2007) documented the impact of rice bran, black gram powder, groundnut cake powder and neem cake powder at three and five percent amendments on the production of pink oyster mushrooms (Pleurotus eous) and discovered that the addition of neem cake at the five percent level considerably enhanced the production of sporophores, with the three percent level. These treatments produced fruiting bodies at the earliest (10.8 to 11 days) than others. All the other amendments, with the exception of rice bran and neem cake powder, had minimal impact on yield growth. Kumar et al. (2012) conducted an assessment involving three distinct substrates, namely wheat straw (WS), maize cob straw (MCS) and lentil straw (LS), either individually or in combinations of WS with MCS and LS at ratios of 1:2, 1:1 and 2:1. Additionally nine supplements were scrutinized including wheat bran, rice bran, corn meal, soybean meal, groundnut cake, pigeon pea powder, lentil powder, mustard cake and neem cake to evaluate their impact on yield performance and biological efficiency across Pleurotus species namely P. sajor-caju, P. florida, P. flabellatus and P. sapidus. Notably, the substrate combination of WS+LS (2:1) exhibited the shortest duration for spawn colonization among all tested Pleurotus species and yielded the highest production of P. sajor-caju (572.80g with 57.30% BE) closely followed by P. florida (554.20g with 55.40% BE). Hasan et al. (2015) investigated the impact of wheat bran supplementation at varying levels (0%, 10%, 20%, 30%, 40% and 50%) on sugarcane bagasse during spawning. They found that 40% supplementation led to the highest mycelium running rate while 30% supplementation resulted in the longest time for pinhead initiation. Additionally, 10% supplementation yielded the longest stipe length while 50% supplementation led to the shortest stipe length and diameter. Furthermore, 30% supplementation resulted in the widest pileus diameter whereas 50% supplementation resulted in the narrowest. Khan et al. (2017) explored the effects of various supplementary materials on oyster mushroom production. They enriched cotton waste with five percent of wheat bran, rice bran and gram flour and examined their impact on spawn running, yield performance and biological efficiency of P. ostreatus. The study concluded that wheat bran supplementation yielded the most favourable results, followed by rice bran and gram flour. The highest biological efficiency at 46.33% was observed in cotton waste supplemented with wheat bran followed by cotton waste supplemented with rice bran at 34.33%. Additionally, the biological efficiency of cotton waste supplemented with gram flour was recorded at 32.33%, while the lowest efficiency was noted in simple cotton waste substrate at 30.00%. Tesfay et al. (2020) evaluated the suitability of waste paper supplemented with wheat bran and maize stalk for oyster mushroom culture. 6 Various concentrations (0%, 25% and 50%) of wheat bran and maize stalk were added to waste paper. It revealed that mixture of 50% waste paper with 25% cornstalk and 25% wheat bran produced better results, with a mean fruiting body weight of 26.20 ± 19.36 g, a pileus diameter of 7.90 ± 2.66 cm, a total output of 646.4 ± 273.1 g and a biological efficiency of 646.4 ± 273 %. In contrast, a lower biological efficiency (17.92 ± 81.95 %) was obtained with waste paper (100%). Furthermore, the maximum mean stalk length (3.88 ± 0.32 cm) was found in waste paper (50%) coupled with maize stalk (50%) supplementation. Sharma et al. (2023) conducted the research on application of substrate supplements to the yield of P. ostreatus var. florida where substrate consisted of sawdust, straw (Rice and Wheat) and supplements included wheat bran, gram flour, maize flour, wheat flour, gram flour, soyabean flour. The most substantial yield was achieved in Wheat Straw (60%), Rice Straw (34%) and Gram Flour (2% of the substrate), resulting in harvests of 452.0 g and 453.0 g. 2.3 Effect of substrate pasteurization Badshah et al. (1992) conducted their research by cultivating P. ostreatus on wheat straw, sugarcane bagasse, corn cobs and sawdust by blending 120-130 g of spawn with 2 kg of substrate and depositing the mixture in sterilized polyethylene bags, which were then placed in darkness at 25°C for 2-3 weeks. Mature fruiting bodies were gathered with yields of 49.8 g/2 kg substrate (sawdust), 432.8 g/2 kg substrate (wheat straw), whereas the control group only yielded 18.5 g/2 kg substrate. Singh et al. (2003) conducted experiments where they cultivated P. sajor-caju using various agricultural residues (such as soybean, paddy, wheat and maize straw) as well as forest by-products (including coir and pine needles) and prior to its cultivation, all substrates were subjected to hot water treatment. Among these substrates, paddy straw exhibited the highest yield at 356.6 g/500g dry weight while pine needles showed a different outcome. The degradation process of these substrates involved the activity of crucial biodegrading enzymes like ligninase, hemicellulase, cellulase, xylanase and β-glucosidase, which are produced by P. sajor-caju. Ali et al. (2007) examined the effects of several pasteurization techniques on cotton waste substrate by subjecting it to hot-water treatment, steam pasteurization and chemical sterilisation with formalin. Of the individual species examined, P. florida, P. pulmonarius and P. ostreatus showed the best results from steam pasteurisation. Akhter et al. (2017) used rice straw that had been heated at 60°C, 80°C and 100°C to study the effects of hot water treatment on P. ostreatus yield, yield characteristics and contamination. Every heat treatment was maintained for one, two and three hours and the outcomes were contrasted with untreated controls (no hot water treatment). The contamination of rice straw was more prevalent in 60°C treated packets compared to non- treated spawn packages. A three-hour heat treatment at 80°C produced better results, with a BE of 57.44%, an economic yield of 280 g/kg, the greatest average pileus diameter (5.0 cm) and the largest average stipe length (3.7 cm). 7  Shrestha et al. (2021) examined the impact of various substrate sterlisation techniques (chemical, steam and hot water) on performance of P. ostreatus and concluded that steam sterlisation was better than rest two. Jeet et al. (2022) evaluated the effect of different substrate pasteurisation techniques and cultivation methods on P. florida and P. sajor-caju. The hot water treatment of wheat straw at 70°C for 20 minutes was determined to be superior to all alternative treatments. Grimm et al. (2024) examined the effects of various pasteurization and sterilization methodologies on the development and output of oyster mushrooms. Their findings indicated that employing autoclaving for substrate sterilization led to a substantial enhancement in dry yields, with increase of up to 50% compared to pasteurization techniques and regarding fresh yield, hot water pasteurization demonstrated comparable efficacy to autoclaving and proved notably superior to alternative pasteurization techniques 2.4 Yield and yield attributing characters Jiskani et al. (2003) assessed that when paddy straw in conjunction with wheat straw, sorghum straw, cotton boll locules, sugarcane debris and banana leaves was used for oyster mushroom cultivation, the pinheads initially emerged 33 days later, after spawning on a blend of soybean and paddy straw. Using paddy straw and cotton boll locules, the shortest time required for fruiting body maturity was found to be 4.25 days following pinhead development. Periasamy and Natarajan (2003) in their research study exhibited the stipe dimensions variability among the recombinant strains of P. djamor. The highest recorded lengths and widths were 2.8 cm and 1.3 cm, respectively, while the minimum measurements were 0.7 cm and 0.6 cm, respectively, the average dimensions of stipe length and width varied across different strains. Shah et al. (2004) carried out an investigation of the growing of oyster mushrooms on various substrates and analysed that a pinhead of P. ostreatus needed 24.0 days on wheat straw, 30.3 days on Wheat straw + leaves, 25.3 days on Saw + wheat straw, 29.3 days on Sawdust + leaves but sawdust and leaves required 24.3 days and 30.3 days, respectively. Iqbal et al. (2005) assessed the yield performance of oyster mushrooms on different substrates including wheat straw, paddy straw, cotton waste and various agricultural residues and indicated that the colonization period for spawn on wheat straw, paddy straw and cotton waste was 43 days, 41.3 days and 30.3 days, respectively and for pinhead 46.3 days for wheat straw, 45.3 days for paddy straw and 32 days for cotton trash were needed. The results of the period for harvesting of P. sajor-caju needed 50.7 days on wheat straw, 50.0 days for paddy straw and 37.7 days for cotton waste. Mandhare et al. (2007) made several findings during the study on the productivity and nutritional value of Pleurotus spp. on paddy straw and its impact on nutritional indices of wasted straw. They detected that the stipe length of P. eous on paddy straw was 1.97 cm. Singh et al. (2010) examined the combining of locally sourced substrates with optimal spawn rates on yields under the natural condition of western Uttar Pradesh. They found that using paddy straw with a three per cent spawn rate resulted in the most favourable 8 substrate, as evidenced by the efficient completion of spawn run and high bio-efficiency. Shukla and Jaitley (2011) conducted the experiment with wheat spawn across five distinct oyster species. Notably, P. sajor-caju exhibited a swift completion of spawn run in a mere 10.5 days, whereas counterparts like P. florida and P. djamor required 13 and 13.5 days, respectively. Alongwith it, P. sajor-caju had stipe lengths of 4.7 cm, pileus lengths of 6.42 cm and 7.25 cm. Singh et al. (2012) raised the spawn of five distinct strains of P. florida and P. sajor-caju on wheat grains to measure the length of time for spawn development. The duration of spawn development ranged from 10 to 17.7 days in case of P. florida strains, but P. sajor-caju strains took up to 9.7 to 18.3 days. Abdullah et al. (2013) employed liquid spawn for the cultivation of P. pulmonarius and observed its superior capacity to colonize sterile rubber wood sawdust in contrast to the conventional grain spawn method. Jatwa et al. (2016) elucidated that the greatest mean pileus diameter (9.3 cm, 12 cm and 12 cm) was achieved for P. florida, P. eous and P. sajor-caju, respectively when cultivated on paddy straw substrate. Conversely, the smallest mean pileus diameter (3.66 cm) was observed when cultivated on combinations of paddy straw + sorghum straw, 2.33 cm in wheat straw + sorghum straw and 4 cm in wheat straw + soybean straw for P. florida, P. eous and P. sajor- caju, respectively across various substrates. Satpal (2017) undertook an experimental investigation to determine the optimal substrate for the cultivation of P. djamor. Wheat straw, paddy straw and chickpea straw were employed individually and in a combined manner with each other at a ratio of 1:1 (w/w) for the cultivation of P. djamor. The most significant dimensions of pileus length and width (9.0 cm and 9.67 cm, respectively) were observed when wheat straw substrate was utilised. In case of harvesting, it was found that wheat straw possessed the first harvesting occurring at 30.0 days and cropping period lasted 61.0 days. Mandaviya et al. (2018) revealed in their study on growth and yield performance of pink oyster mushroom on different substrates that wheat straw was the next thickest pink oyster mushroom stipe (6.21 cm) whereas sugarcane bagasse having the maximum thickness at 6.53 cm. The thickness of the stipe was 5.43 cm on sorghum straw and 5.99 cm on paddy straw, whereas corn straw produced the lowest stipe thickness (5.04 cm). Neupane et al. (2018) observed that wheat straw substrate exhibited the most substantial cap diameter (9.87 cm) followed by banana leaves (9.17 cm), rice straw (8.96 cm), a combination of rice and wheat straw (8.11) cm and sawdust presented the smallest diameter (7.14 cm). Patar et al. (2018) scrutinized the growth dynamics and yield potential of two oyster mushroom species, P. florida and P. sajor-caju, on wheat substrate. The pinhead emergence showed an early flourish, with P. florida revealing its presence in a swift 15 days, while P. sajor-caju trailed slightly behind at 18 days. Sitaula et al. (2018) examined the effects on growth and yield performance of four different substrates viz. 100% paddy straw, maize cob + paddy straw (1:1), sugarcane bagasse + paddy straw (1:1) and sawdust + paddy straw (1:1) regarding P. ostreatus. The study revealed that the maize + paddy straw combination required the longest 9 time for the spawn run (20.50 days), whereas the paddy straw combination required the shortest time (18.25 days). Dubey et al. (2019) investigated the impact of various substrates on the performance of P. sajor-caju and the substrates tested for cultivation included rice straw (T1), wheat straw (T2), banana leaves (T3) and sugarcane bagasse (T4), the highest stipe length (4.9 cm) was recorded with rice straw followed by banana leaves, wheat straw and sugarcane bagasse at 4.3 cm, 3.7 cm and 3.3 cm, respectively. Shroff (2019) investigated five oyster mushroom strains for different growth attributes and reported that pin head commencement was 20.7 days in strain R-13-13-5, whereas other strains R-51, R-1313-04 and R-13-1303 required 23.50 days, 24.17 days and 24.67 days, respectively. The strain R-13- 13-07 required the highest number of days for pin head initiation (25.0 days). Soni et al. (2020) observed varying harvesting durations flush of P. florida on different substrates. Wheat straw exhibited the swiftest yield at 22 days, followed by mustard straw at 24 days, rice straw and a rice-wheat straw mix at 27 and 26 days, respectively. Maize straw required the longest period at 34 days. For the second flush, durations ranged from 35 to 42 days, maize straw required the longest time (42 days) and wheat straw and mustard straw needed the least (35 days), followed by rice straw at 36 days. The harvesting of third flush ranged from 47 to 50 days, with wheat straw and a rice-wheat straw combination taking the shortest time at 47 days. Vanmathi and Lakshmi (2020) investigated the effects of several substrates, including sugarcane bagasse, sawdust and paddy straw, on the development and yield of P. ostreatus and observed that the paddy straw had the largest pileus diameter (6.41 ± 0.60 cm) followed by sawdust (5.50 ± 0.06 cm) and sugarcane bagasse (4.48 ± 1.41 cm). Muswati et al. (2021) determined the influence of substrate mixing on oyster mushroom development and yield. They used seven substrates viz. (Trt1) cotton waste, (Trt2) wheat straw, (Trt3) crushed baobab fruit shells, 100%, (Trt4) cotton husk + wheat straw + crushed baobab fruit shells, 1:1:1, (Trt5) baobab fruit shells + cotton husks, 1:1, (Trt6) baobab fruit shells + wheat straw, 1:1 and (Trt7) cotton waste + wheat straw, 1:1. The treatment Trt1 produced the biggest cap diameter (5.5 cm) and it was followed by treatment Trt5 (4.8 cm). The cap diameters were 4.1 cm, 4.2 cm and 4.3 cm in Trt2, Trt7 and Trt3 as compared to mean (4.7 cm). In case of stipe thickness, cotton husk, wheat straw and crushed baobab fruit shells produced the maximum stipe girth (5.40 cm) while highest cap diameter of 5.22 cm was exhibited by cotton waste. Sen et al. (2022) assessed the production and growth behaviour of blue oyster mushrooms based on different bed materials and spawn while considering the five substrates and found that bagasse from sugarcane took the least time (16.50 days) for the spawn run, while the longest time (23.75 days) was seen in groundnut hulls. 2.5 Benefit Cost Ratio Jadhav (1996) investigated the annual yield performance of various Pleurotus species, including P. sajor-caju, P. florida, P. citrinopileatus and P. ostreatus utilizing wheat straw as the substrate. Among these species, P. florida and P. ostreatus demonstrated the 10 highest yields, reaching 721 g/kg and 434 g/kg, respectively. Notably, P. florida exhibited the highest yield compared to the other species tested. Ahmed et al. (2016) conducted an assessment of yield and nutritional composition across various oyster mushroom species. Their findings indicated that P. ostreatus exhibited the highest biological yield, recording (278 g) productivity at 55% and biological efficiency of 96%. Arathy and Das (2016) analysed the relevancy of locally accessible materials as substrates by raising pink oyster mushrooms on various substrates. These consist of sawdust, coir pith compost, banana pseudo stem, sawdust, sugarcane bagasse and coconut leaf shreds. Banana pseudo stems had the highest biological efficiency of 105.8 % followed by 92.6% of paddy straw. Hossain (2017) documented the superior yield of P. sajor-caju, amounting to 803.0 g/kg, when cultivated on paddy straw, with wheat straw yielding slightly lower at 601.0 g/kg. Gorai and Sharma (2018) in their research findings on effect of different substrates on yield potential of Pleurotus spp. in controlled mushroom house conditions utilizing three distinct substrates: paddy straw, a combination of paddy straw and sugarcane bagasse in a 1:1 ratio and sugarcane bagasse alone. They observed that the highest levels of biological efficiency were attained with paddy straw as the substrate ranging from 93.2% to 84.6%. Subsequently, the blend of paddy straw and sugarcane bagasse demonstrated moderate efficiency, falling within the range of 80.40% to 75.60%. Conversely, the exclusive use of sugarcane bagasse yielded the lowest biological efficiency ranging from 67.60% to 41.60%. Bhandaria et al. (2021) conducted a benefit-cost ratio analysis of the production of P. ostreatus mushrooms on various substrates like rice straw, khar, banana leaves and tree leaves. They recorded a total output of 1230 g, 230 g, 211 g and 480 g, respectively. The benefit cost ratio was calculated respectively to be 2.47, 0.46, 0.42 and 0.96 under rice straw, banana leaves, tree leaves and khar, respectively. They concluded that rice straw had a high benefit-cost ratio for Pleurotus species. Chandra and Saumya (2021) evaluated yield of P. djamor by utilizing various agricultural residues and achieved the highest total yield from wheat straw mushroom bags (941.13 g per bag). It was followed by a mixture substrate consisting of wheat straw with paddy straw (1:1), wheat straw with sawdust (1:1), wheat straw with paddy straw and tamarind leaves (1:1:1) and wheat straw with paddy straw and Bermuda grass (1:1:1) yielded at 811.8g, 628.3g, 533.2g and 473.9g, respectively. Kushwaha and Kushwaha (2023) studied the yield performance of six strains of Pleurotus species on wheat straw as the substrate. The highest yield at 414.4 g per bag was achieved by the PL-3 strain, followed by 399.8 g per bag in PL-2 strain and 379.2 g per bag in PL-5 strain. Conversely, the lowest yield was observed in the PL-4 strain, producing 192.6 g per bag. Additionally, they also observed that PL-3 exhibited the highest biological efficiency at 69.07%, followed by at 66.6% in PL-2 and 63% in PL-5 strain. 11 CHAPTER-III MATERIALS AND METHODS The investigation entitled “Evaluation of biological efficiency of Pleurotus spp. on different agro-wastes in Haryana” was carried out in the Mushroom Technology Laboratory, Department of Plant Pathology, CCS Haryana Agricultural University, Hisar, during 2023-24. Following heads have been used to describe the materials and methods during the investigation. 3.1 Materials 3.1.1 Equipments The investigation utilized borosilicate glasswares including test tubes, petri plates, beakers, conical flasks, volumetric flasks, glass rods, measuring cylinders, pipettes, non- absorbent cotton, funnels, etc. These items were procured from the Department of Plant Pathology, College of Agriculture, CCS HAU, Hisar. 3.1.2 Chemicals Various experiments needed dextrose, agar-agar, calcium carbonate, formaldehyde solution, carbendazim and absolute alcohol, etc. 3.1.3 Pure Culture Pleurotus djamor and Pleurotus pulmonarius pure cultures were utilized for the reported investigation. P. djamor and P. pulmonarius cultures were procured from Mushroom Technology Laboratory, Department of Plant Pathology, CCS Haryana Agricultural University, Hisar, during 2023-24. 3.1.4 Miscellaneous Inoculating needle, rubber bands, gas burner, forceps, non-absorbent cotton, calcium sulphate, calcium carbonate, wheat grains, wheat straw (WS), paddy straw (PS), mustard straw (MS), saw dust (SD), aluminium foil, polypropylene sheets, polypropylene bags etc. and wheat bran (WB), Molasses (M) were required for supplementation of substrates. 3.2 METHODS 3.2.1 Sterilization of Glassware Glasswares were sterilized in hot air oven at 180°C for 1 h. 3.2.2 Sterilization of Media Media and wheat grains were sterilized in an autoclave at 121°C temperature (15 psi) for 30 min and 2 h, respectively. The autoclavable polypropylene bags were used. 3.2.3 Sterilization of Other Equipments 12  Hot air oven and autoclave were used for the sterilization process. Inoculating needle, cork-borer, forceps, etc. were initially dipped in spirit or in 70 % ethanol and then sterilized on flame on spirit lamp until got red and then allowed to cool before use. 3.2.4 Sterilization of Substrates The substrates were chemically treated by soaking in solution of formaldehyde (1250 ppm), carbendazim (75 ppm) for overnight. On the next day, after decanting the excess water on clean sloppy surface lined with polypropylene sheet, the substrates were spread on polythene sheet for about 4 h until substrates are having 65-70 per cent moisture. It was used for spawning purpose. 3.2.5 Maintenance of Culture The pure cultures were maintained on Potato Dextrose Agar (PDA). Fungus was sub cultured at an interval of 8 days followed by its storage under controlled conditions. The cultures were further stored at 4°C and revived once a month. Seven days old mycelial cultures were used for spawn preparation. 3.2.6 Spawn Preparation 3.2.6.1 Master Spawn Preparation For the preparation of spawn, clean, bold and healthy wheat grains were employed. To eliminate potential contaminants, the grains underwent a thorough rinsing with water. Following this, the grains were subjected to boiling in water for a duration of 20 minutes to soften them. After draining off excess water, the grains were spread out on polythene sheet for cooling. Subsequently, the grains were supplemented with CaCO3 and CaSO4 @ 0.2% and 5% (w/w basis), aiming to prevent clump formation and to maintain the grains' pH 7.5. The prepared substrate was then filled into 500 ml glucose/milk bottles up to 2/3rd of the volume and sealed with non-absorbent cotton. These bottles were subsequently sterilized by autoclaving at 121°C for 2 h under a pressure of 15 psi. Upon sterilization, the bottles were allowed to cool and aseptically pure culture was introduced into them. The inoculated bottles were then incubated at 25°C in a BOD incubator for a period of 14 days to facilitate mycelial proliferation. 3.2.6.2 Commercial Spawn Preparation Commercial spawn was produced within durable polypropylene bags resistant to heat. Each bag underwent double sealing and was filled with 1 kg of wheat grain. The processing and filling of wheat grains adhered to the methodology employed in the preparation of master spawn. Following filling, the bags were subjected to sterilization in an autoclave at a pressure of 15 psi for 2 h. Subsequently, the bags were left to cool overnight before being aseptically inoculated with 20 g of master spawn. The inoculated bags were agitated to ensure homogeneous mixing of the inoculum with the grains and were then placed in a BOD incubator set at 25 ± 1°C for a period of 14 days to facilitate mycelial proliferation. 13 3.2.7 Chemically pasteurized substrates and their supplementation on growth characteristics as well as yield of P. djamor and P. pulmonarius. The preparation of substrates for wheat straw, mustard straw, paddy straw and sawdust involved chemical treatment. Each substrate, weighing 10 kg, was soaked overnight in a solution of formaldehyde (1250 ppm), carbendazim 50 WP (75 ppm). Additionally, Wheat bran (WB) was supplemented at 10 per cent of the dry weight of the substrate, while Molasses (M) was added at 2 per cent of the dry weight of the substrate. Sterlization of the substrates was carried out, followed by inoculation with wheat grain-based spawn at a rate of 10 per cent of the dry weight of the substrate. The inoculated substrates were then filled into polythene bags (40 x 60 cm) at a rate of 2.5 kg per bag of wet substrate (with 60 % moisture content). Cultivation of P. djamor and P. pulmonarius was carried out during the months of October 2023 and December 2023, respectively at Mushroom Technology Lab, CCS HAU Hisar and standard agronomic practices were followed. Statistical analysis was conducted using Randomized Block Design (RBD) with three replications for each treatment and each replication consisted of five bags. Observations were recorded for various parameters including time taken for spawn run, time taken for pin head formation (days), time taken for first flush (days), second flush (days), crop duration (days), pileus size (cm), stipe thickness (cm), yield (g/kg dry substrate), disease incidence and insect pest infestation (if any) and benefit-cost ratio. 3.2.7.1 Observations recorded i. Spawn run (days) This was recorded by counting days from the initial filling of bags to the attainment of complete mycelial growth or spawn run within each bag across all treatment groups. ii. Pinhead formation (days) Pinhead formation was observed by recording the time taken in days from the commencement of bag filling to the emergence of pinheads for each treatment. iii. Time for first flush (days) iv. Time for second flush (days) v. Crop duration (days) vi. Pileus size (cm) vii. Stipe thickness (cm) viii. Yield (g/kg dry substrate) This was calculated by weighing of fresh fruiting body each bag during cropping season. 14  ix. Disease incidence and insect pest infestation x. Benefit-Cost Ratio For each treatment under experimentation, benefit-cost ratio was calculated separately Benefit-Cost ratio = Benefit (₹) from yield / Costs (₹) for all inputs Treatment details T1 = WS alone T2 = WS+WB (10%) T3 = WS+M (2%) T4 = MS alone T5 = MS+WB (10%) T6 = MS+M (2%) T7 = PS alone T8 = PS+WB (10%) T9 = PS+M (2%) T10 = SD alone T11 = SD+WB (10%) T12 = SD+M (2%) 3.2.8 Heat pasteurized substrates and their supplementation on growth characteristics as well as yield of P. djamor and P. pulmonarius. The substrate preparation for wheat straw, mustard straw, paddy straw and sawdust involved a heating method. Each substrate, after being soaked overnight in water, underwent heating at 70℃ for a duration of 20 minutes. Wheat bran (WB) was supplemented at 10 per cent of the dry weight of the substrate, while Molasses (M) was added at 2 per cent of the dry weight of the substrate. Pasteurized substrates were then inoculated with wheat grain-based spawn at a rate of 10 per cent of the dry weight of the substrate and filled into polythene bags (40 x 60 cm) at a rate of 2.5 kg per bag of wet substrate (with 60 % moisture content). Cultivation of P. djamor and P. pulmonarius was conducted during the months of October 2023 and December 2023, respectively at Mushroom Technology Lab, CCS HAU Hisar and standard agronomical practices were employed. Statistical analysis was conducted using Randomized Block Design (RBD) with three replications for each treatment and each replication consisted of five bags. Observations were recorded for various parameters including time taken for spawn run (days), time taken for pin head formation (days), time taken for first flush (days), second flush (days) and crop duration (days), pileus size (cm), stipe thickness (cm), yield (g/kg dry substrate), disease incidence and insect pest infestation (if any) and benefit-cost ratio. 15 3.2.8.1 Observations recorded i. Spawn run (days) This was recorded by counting days from the initial filling of bags to the attainment of complete mycelial growth or spawn run within each bag across all treatment groups. ii. Pinhead formation (days) Pinhead formation was observed by recording the time taken in days from the commencement of bag filling to the emergence of pinheads for each treatment. iii. Time for first flush (days) iv. Time for second flush (days) v. Crop duration (days) vi. Pileus size (cm) vii. Stipe thickness (cm) viii. Yield (g/kg dry substrate) This was calculated by weighing of fresh fruiting body of each bag during cropping season. ix. Disease incidence and insect pest infestation x. Benefit-Cost Ratio Treatment details T1 = WS alone T2 = WS+WB (10%) T3 = WS+M (2%) T4 = MS alone T5 = MS+WB (10%) T6 = MS+M (2%) T7 = PS alone T8 = PS+WB (10%) T9 = PS+M (2%) T10 = SD alone T11 = SD+WB (10%) T12 = SD+M (2%) 3.2.9 Cultivation of P. djamor and P. pulmonarius The cultivation of P djamor and P. pulmonarius involved following steps: 3.2.9.1 Substrate preparation The oyster mushroom was cultivated on substrates i.e. paddy straw (Oryza sativa), wheat straw (Triticum aestivum), mustard straw (Brassica juncea.) and sawdust (mixture of Acacia nilotica and Eucalyptus) pasteurized by chemical method and heating method. Polythene bag of size 40 x 60 cm was filled with substrate weighing 2.5 kg. Before 16  (a) Straw chopping (b) Soaking of substrates (c) Soaking of substrates in chemical (d) Cooling of substrates after heating (e) Spreading of substrates After (f) Addition of supplement pasteurization (g) Mixing of spawn Plate 1. (a-g) Substrate prepration and spawning 17 pasteurisation and filling in bags hand-threshed straw free from leafy debris was chopped into lengths of 1.5-2” size using a chaff cutter, packed into gunny bags, and immersed in water containing 1250 ppm formaldehyde and 75 ppm carbendazim 50 WP for overnight and container was covered with a polyethylene sheet. Excess water was removed by spreading the soaked straw on a clean cemented surface. Prior to substrate filling in bags, the moisture content of the substrate 60-65 % was attained. 3.2.9.2 Spawning The pasteurized substrate got ready for filling and spawning in polybags when it cooled down to room temperature and the moisture content was about 60-65 per cent. The spawning was done in neat and clean place. Polybags 40 x 60 cm used for filling the substrate were permitted to create 10 holes of about 1 cm in diameter. In the experiment care was taken to uniform mix the spawn in the substrate and then filled in bags with gentle pressing and the mouth of the poly bag was tied with rubber bands. 3.2.9.3 Cropping After spawning, the bags were kept apart in racks at a distance of 15 cm. The temperature of the mushroom house was maintained between 23-28°C during the cropping period and the relative humidity was maintained at 80-85 per cent by frequent sprays of water on the floor and walls of the mushroom growth house. 3.2.9.4 Picking The matured mushroom was harvested before the margin of cap curls and harvested by twisting it clock wise to uproot from the bag. The harvesting of mushrooms was done carefully so that the young developing pinheads in the vicinity did not become vulnerable to any sort of damage and loss. In a single life cycle of mushroom, three flushes appeared. The harvested mushrooms were trimmed off substrate level and weighed separately for each bag at every harvest. Following the initial harvest, mushroom fruiting persisted at an interval of 7- 10 days for up to two successive flushes. 3.2.9.5 Yield The measurements in various growth parameters and yield (g) of P. djamor and P. pulmonarius was summed up bag wise and then replication wise in all treatments throughout the cropping process. 18 3.2.9.6 Benefit-Cost Ratio The determination of the Pleurotus spp. cultivation expenses per kg of mushroom production was conducted through the aggregation of fixed costs (Includes Mushroom shed) and variable costs (Includes straw, supplements, spawn, polybags, labour, chemicals etc.). Gross income was derived by multiplying the harvested weight of fresh mushrooms (kg) by the market value ₹ per kg of fresh mushrooms. Net income was then ascertained by deducting cost of cultivation from the gross income acquired, subsequently leading to the determination of the benefit-cost ratio [Total returns (₹) / Cost of cultivation (₹)]. 19 CHAPTER-IV RESULTS The present investigation entitled “Evaluation of biological efficiency of Pleurotus spp. on different agro-wastes in Haryana” was carried out in the Mushroom Technology Laboratory, Department of Plant Pathology, CCS Haryana Agricultural University, Hisar, during 2023-24. For better understanding and to cover all the objectives of the study, results are presented under the following eight headings: 4.1 Effect of chemically pasteurized substrates and their supplementation on growth characteristics as well as yield of P. djamor 4.2 Effect of heat pasteurized substrates and their supplementation on growth characteristics as well as yield of P. djamor 4.3 Effect of chemically pasteurized substrates and their supplementation on growth characteristics as well as yield of P. pulmonarius 4.4 Effect of heat pasteurized substrates and their supplementation on growth characteristics as well as yield of P. pulmonarius 4.5 Evaluation of BC ratio upon cultivation of P. djamor on chemical pasteurised substrates and their enrichment with substrates 4.6 Evaluation of BC ratio upon cultivation of P. djamor on heat pasteurised substrates and their enrichment with substrates 4.7 Evaluation of BC ratio upon cultivation of P. pulmonarius on chemical pasteurised substrates and their enrichment with substrates 4.8 Evaluation of BC ratio upon cultivation of P. pulmonarius on heat pasteurised substrates and their enrichment with substrates To ensure the study, results are presented clearly and effectively, detailed sections are provided below: 4.1 Effect of chemically pasteurized substrates and their supplementation on growth characteristics as well as yield of P. djamor 4.1.1 Effect of chemically pasteurized substrates and their supplementation on crop cycle of P. djamor The results presented in Table 4.1 revealed the data concerning the effect of chemically pasteurized substrates like wheat straw, mustard straw, paddy straw, saw dust and their supplementation i.e., wheat bran and molasses on crop cycle of P. djamor. The observations of spawn run duration were recorded upon full colonization of the mushroom substrates by the mycelium. The results on spawn run clearly showed that the time taken for spawn run of the different treatments varied from 17.3 to 35.3 days. Among the 20 different substrates supplemented with wheat bran and molasses exhibited shorter spawn run durations as compared to their respective substrates without supplementation. It was observed that wheat straw supplemented with wheat bran exhibited significantly shortest period of 17.3 days for colonisation of substrates as compared to other treatments. It was followed by 18.7 days in wheat straw enriched with molasses. The paddy straw substrates showed an intermediate spawn run durations with 23.0 days for paddy straw alone, 20.3 days for paddy straw with wheat bran and 21.7 days for paddy straw with molasses. In mustard straw substrates, the spawn run duration was noticeably longer with mustard straw alone requiring 31.3 days, which was reduced to 27.30 days when supplemented with wheat bran and 29.0 days with molasses. The saw dust required longest period of 35.3 days to complete spawn run. The duration of pinhead formation also varied significantly among different treatments which ranged from 21.3 to 42.7 days The wheat straw + wheat bran which was chemically pasteurised had significantly shortest duration for pinhead formation at 21.3 days followed by wheat straw enriched with molasses at 21.7 days and then by wheat straw alone at 23.3 days. The paddy straw also demonstrated reduced pinhead formation days with supplementation, it was 24.3 days with wheat bran and 26.3 days with molasses as compared to 28.7 days without supplementation. The mustard straw exhibited longer pinhead formation durations with 37.0 days without supplementation and it reduced to 32.7 days with wheat bran and 36.0 days with molasses supplementation. The saw dust without supplements required the significantly longest period at 42.7 days for pinhead formation. Amidst the different treatments, days required for first flush of harvesting were significantly lowest in the wheat straw + wheat bran chemically pasteurised i.e. 27.3 days, followed by 28.3 days in wheat straw enriched with molasses. The paddy straw also demonstrated significantly reduced time to the first flush with supplementation i.e., 31.3 days with wheat bran and 32.7 days with molasses as compared to 34.7 days in paddy straw alone. The mustard straw substrates required significantly more time for the first flush of harvesting with duration ranged from 40.0 days with wheat bran to 43.0 days without supplementation and upto 42.7 days with molasses. It was noticed that maximum number of 47.3 days were required for first harvest in sawdust chemically pasteurised and it was significantly longest period as compared to the other treatments. Data presented in Table 4.1 revealed that the number of days required for the second harvest were calculated by counting the days from spawning to the second harvest of mushroom. Among the different treatments, the second flush of P. djamor was completed within 37.0 to 55.3 days in different treatments of Table 4.1. It was significantly lowest at 37.0 days in wheat substrate supplemented with wheat bran and differed significantly among other treatments, followed by wheat substrate supplemented with molasses at 40.0 days The 21 paddy straw exhibited lesser times to the second flush with supplementation, ranged from 42.0 days with wheat bran to 45.3 days without supplementation and 43.0 days with molasses. The mustard straw substrates exhibited significantly longer durations to the second flush ranged from 50.0 days with wheat bran to 51.0 days. The longest period of second flush of harvest was found at 55.3 days in sawdust. The wheat straw supplemented with wheat bran exhibited significantly shortest crop duration of 52.0 days as compared to other treatments. It was followed by 52.3 days in wheat straw enriched with molasses. The paddy straw also showed reduced crop durations with supplementation, ranged from 53.7 days with wheat bran to 55.7 days without supplementation and 54.7 days with molasses. The mustard straw substrates required longer durations for crop completion, ranged from 58.3 days with wheat bran to 59.3 days without supplementation. Meanwhile saw dust required longest crop duration of 63.3 days to complete spawn run and it was significantly longest crop duration as compared to the other treatments (Table 4.1). Table 4.1: Effect of chemically pasteurized substrates and their supplementation on crop cycle of P. djamor Spawn Pinhead 1st 2nd Crop Treatment details run* formation* flush* flush* duration* (days) (days) (days) (days) (days) Wheat Straw 19.3 23.3 30.0 41.5 52.7 Wheat Straw + Wheat Bran 17.3 21.3 27.3 37.0 52.0 Wheat Straw + Molasses 18.7 21.7 28.3 40.0 52.3 Mustard Straw 31.3 37.0 43.0 51.0 59.3 Mustard Straw + Wheat Bran 27.3 32.7 40.0 50.0 58.3 Mustard Straw + Molasses 29.0 36.0 42.7 50.7 59.0 Paddy Straw 23.0 28.7 34.7 45.3 55.7 Paddy Straw + Wheat Bran 20.3 24.3 31.3 42.0 53.7 Paddy Straw + Molasses 21.7 26.3 32.7 43.0 54.7 Saw Dust 35.3 42.7 47.3 55.3 63.3 Saw Dust + Wheat Bran 32.7 38.7 44.0 52.3 60.0 Saw Dust + Molasses 34.7 40.0 47.0 54.3 62.3 CD at 5 % level 2.03 1.19 1.29 2.31 1.92 *Average of three replications and five bags per replication 22 4.1.2 Effect of chemical pasteurised substrates and their supplementation on growth characteristics and yield of P. djamor. The Table 4.2 presents results from an experiment examining the effect of chemical pasteurised substrates and their supplementation on growth characteristics like pileus size (cm), stipe thickness (cm) and yield (g/kg of dry weight of substrate) of P. djamor. The substrates supplemented with wheat bran or molasses exhibited notable improvements in both pileus size and stipe thickness as compared to their unsupplemented counterparts. The pileus size of fruit bodies of P. djamor was ranged from 3.5 to 6.7 cm among different treatments. The maximum pileus size (6.7 cm) was noticed on chemically pasteurised wheat substrate enriched with wheat bran which was highly significant than other treatments, followed by pileus size in wheat straw enriched with molasses (6.6 cm). The paddy straw substrate treatments displayed varying pileus sizes, with values ranged from 5.7 cm without supplementation to 6.2 cm with wheat bran and 5.9 cm with molasses. The mustard straw substrates showed smaller pileus size ranged from 5.2 cm without supplementation to 5.6 cm with wheat bran and 5.5 cm with molasses. The pileus size was significantly minimum at 3.5 cm in sawdust chemical pasteurised. Data provided in Table 4.2 revealed that among different treatments, highest mean stipe thickness of 1.97 cm was noticed on wheat substrate enriched with wheat bran. It was significantly more as compared to all other treatments. It was followed by wheat straw supplemented with molasses exhibiting a stipe thickness of 1.87 cm. The paddy straw treatments demonstrated varying stipe thicknesses, with values ranged from 1.65 cm without supplementation to 1.73 cm with wheat bran and 1.70 cm with molasses. The mustard straw substrates exhibited thinner stipes ranged from 1.47 cm without supplementation to 1.53 cm with wheat bran and 1.50 cm with molasses. The minimum stipe thickness of 1.41 cm was observed in sawdust. The yield was calculated replication wise by adding the fresh weight of all of the three harvests. The total yield ranged from 450.0 to 710.0 g per kg dry substrate on different treatments (Table 4.2). The highest yield (710.0 g/kg dry substrate) was recorded on chemically pasteurised wheat straw supplemented with wheat bran which was significantly higher than all the other treatments. It was followed by wheat straw enriched with molasses (682.3 g/kg dry substrate) and wheat straw alone (670.0 g/kg). The paddy straw based treatments demonstrated improved yields with values ranged from 525.0 g/kg without supplementation to 595.0 g/kg with wheat bran and 560.0 g/kg with molasses. The yield of mustard straw based substrates ranged from 475.0 g/kg to 485.0 g/kg, which was comparable to the yields of sawdust based treatments ranged from 450.0 g/kg to 468.0 g/kg. The lowest yield was obtained on chemically pasteurised sawdust (450.0 g/kg), which was not significantly better than all other treatments. 23  4.2 Effect of heat pasteurized substrates and their supplementation on growth characteristics as well as yield of P. djamor Table 4.2 Effect of chemical pasteurised substrates and their supplementation on growth characteristics and yield of P. djamor Yield* Pileus size* Stipe thickness* Treatment details (g/kg of dry weight) of (cm) (cm) substrate) Wheat Straw 6.4 1.77 670.0 Wheat Straw + Wheat Bran 6.7 1.97 710.0 Wheat Straw + Molasses 6.6 1.87 682.3 Mustard Straw 5.2 1.47 475.0 Mustard Straw + Wheat Bran 5.6 1.53 485.0 Mustard Straw + Molasses 5.5 1.50 480.0 Paddy Straw 5.7 1.65 525.0 Paddy Straw + Wheat Bran 6.2 1.73 595.0 Paddy Straw + Molasses 5.9 1.70 560.0 Saw Dust 3.5 1.41 450.0 Saw Dust + Wheat Bran 4.0 1.45 470.0 Saw Dust + Molasses 3.8 1.43 468.0 CD at 5 % level 0.20 0.15 9.86 *Average of three replications and five bags per replication 4.2.1 Effect of heat pasteurised substrates and their supplementation on crop cycle of P. djamor Table 4.3 depicted the effect of heat-pasteurized substrates and their supplementation on the crop cycle of P. djamor, involving spawn run, pinhead formation, time for first flush, time for second flush and crop duration. The results detailed in Table 4.3 illustrated varying spawn run durations across treatments, ranged from 23.0 to 39.3 days. The substrates supplemented with wheat bran or molasses exhibited shorter spawn run periods as compared to their unsupplemented counterparts. The heat pasteurised wheat straw supplemented with wheat bran achieved the significantly shortest spawn run duration of 23.0 days. It was followed by wheat straw with molasses at 23.3 days. The heat pasteurised paddy straw treatments displayed the intermediate spawn run durations i.e. 26.0 days with wheat bran and 28.7 days with paddy straw alone. The mustard straw substrates showed longer spawn run durations; it was 30.7 days with wheat bran and 32.3 days with molasses as compared to 34.7 days with mustard straw alone. Meanwhile, sawdust showed significantly longest spawn run period of 39.3 days for completion of spawn run. 24 The Table 4.3 presented the effect of heat-pasteurized substrates and their supplementation on pinhead formation of P. djamor. Among the treatments, heat-pasteurized wheat straw supplemented with wheat bran had significantly shortest duration for pinhead formation at 26.3 days. It was followed by wheat straw with molasses at 27.0 days. The paddy straw treatments demonstrated reduced times to pinhead formation with supplementation i.e. 30.0 days with wheat bran and 32.7 days with molasses compared to 34.3 days in paddy straw alone. The mustard straw substrates exhibited longer pinhead formation durations i.e., 35.7 days with wheat bran and 37.3 days with molasses as compared to 39.7 days in mustard straw without supplementation. In contrast, substrates such as sawdust without supplements demonstrated significantly longest duration for pinhead formation at 44.7 days. Among the different treatments, days required for first flush of harvesting were significantly lowest in the heat-pasteurized wheat straw enriched with wheat bran i.e. 33.0 days and followed by wheat straw enriched with molasses at 35.0 days. The paddy straw based treatments also demonstrated shorter times to first flush of harvest with supplementation i.e. 37.7 days with wheat bran and 39.0 days with molasses compared to 40.3 days in paddy straw alone. The mustard straw substrates exhibited prolonged durations to first flush, ranged from 43.0 days with wheat bran to 44.7 days without supplementation. It was noticed that maximum number of days (49.3 days) were required for first harvest in heat- pasteurized sawdust and it was significantly longest period for first flush compared to the other treatments. Data presented in Table 4.3 revealed that among the different treatments, the second flush of P. djamor was completed within durations of 43.0 to 56.3 days in different treatments. It was significantly lowest at 43.0 days in wheat substrate supplemented with wheat bran which differed significantly among other treatments. It was followed by wheat substrate supplemented with molasses at 44.0 days. The paddy straw based treatments exhibited slightly shorter duration to the second flush with supplementation i.e. 46.0 days with wheat bran and 46.7 days with molasses as compared to 47.0 days in paddy straw alone. The mustard straw substrates exhibited significantly longer durations to the second flush ranged from 51.0 days with wheat bran to 52.7 days in mustard straw alone. The longest period of second flush of harvest was found at 53.7 days in sawdust. The crop duration also varied significantly among treatments. The heat-pasteurized wheat straw supplemented with wheat bran exhibited the shortest crop duration of 54.3 days, followed by wheat straw enriched with molasses at 55.0 days. The paddy straw treatments demonstrated durations of 56.2 days with wheat bran and 56.7 days with molasses as compared to 57.0 days in paddy straw alone, indicating moderate variations. Meanwhile mustard straw substrates exhibited longer crop durations, ranged from 58.0 days with wheat 25 bran to 59.9 days without supplementation as compared to wheat and paddy straw based treatments. The significantly longest crop duration exhibited by sawdust at 68.0 days (Table 4.3). Table 4.3: Effect of heat pasteurised substrates and their supplementation on crop cycle of P. djamor Spawn Pinhead 1st 2nd Crop Treatment details run* formation* flush* flush* duration* (days) (days) (days) (days) (days) Wheat Straw 24.3 29.3 36.0 44.7 55.9 Wheat Straw + Wheat Bran 23.0 26.3 33.0 43.0 54.3 Wheat Straw + Molasses 23.3 27.0 35.0 44.0 55.0 Mustard Straw 34.7 39.7 44.7 52.7 60.0 Mustard Straw + Wheat Bran 30.7 35.7 43.0 51.0 58.0 Mustard Straw + Molasses 32.3 37.3 44.0 52.0 59.0 Paddy Straw 28.7 34.3 40.3 47.0 57.0 Paddy Straw + Wheat Bran 26.0 30.0 37.7 46.0 56.2 Paddy Straw + Molasses 28.0 32.7 39.0 46.7 56.7 Saw Dust 39.3 44.7 49.3 56.3 68.0 Saw Dust + Wheat Bran 36.3 41.3 46.7 53.7 65.3 Saw Dust + Molasses 37.7 42.7 48.0 55.0 66.7 CD at 5 % level 0.91 1.16 1.45 1.34 2.62 *Average of three replications and five bags per replication 4.2.2 Effect of heat pasteurised substrates and their supplementation on growth characteristics and yield of P. djamor The Table 4.4 presents results from an experiment examining the effect of heat pasteurised substrates and their supplementation on growth characteristics like pileus size (cm), stipe thickness (cm) and yield (g/kg of dry weight of substrate) of P. djamor. The pileus size of fruit bodies of P. djamor ranged from 4.1 to 8.3 cm among different treatments. The maximum pileus size of 8.3 cm was observed on heat pasteurised wheat substrate enriched with wheat bran which was highly significant than others, followed by pileus size in wheat straw enriched with molasses of 8.1 cm. The paddy straw based treatments showed slightly larger pileus sizes as compared to mustard and sawdust based substrates and it ranged from 7.5 cm with wheat bran to 7.2 cm without supplementation and 7.4 cm with molasses. The pileus size was significantly minimum at 4.1 cm in sawdust heat pasteurised. 26  (a) Pinhead formation (b) Fruiting body formation (c) Ready for harvesting (d) Weighing of mushroom Plate 2. (a-d) Fruiting in P. djamor Data provided in Table 4.4, revealed that among different treatments, highest stipe thickness of 2.13 cm was noticed on wheat substrate enriched with wheat bran. It was significantly more as compared to all other treatments, followed by wheat straw supplemented with molasses exhibiting a stipe thickness of 1.93 cm and 1.90 cm for wheat straw without enrichment. The paddy straw with wheat bran comprised stipe thickness of 1.80 cm showing moderate variations with other treatments. The mustard straw substrates exhibited thinner stipes, thickness ranged from 1.68 cm with wheat bran to 1.60 cm without supplementation and 1.63 cm with molasses. The minimum stipe thickness of 1.43 cm was recorded in sawdust. The total yield ranged from 392.4 to 755.0 g per kg dry substrate on different treatments (Table 4.4). The highest yield (755.0 g/kg dry substrate) was recorded on heat pasteurised wheat straw supplemented with wheat bran which was significantly higher than other treatments. It was followed by wheat straw enriched with molasses (736.4 g/kg dry 27 substrate). The yield of 693.8 g/kg was obtained for wheat straw alone. The paddy with wheat bran yielded 655.0 g/kg and paddy straw with molasses yielded 635.0 g/kg, indicating moderate variations. The mustard substrate). The yield of 693.8 g/kg was obtained for wheat straw alone. The paddy with wheat bran yielded 655.0 g/kg and paddy straw with molasses yielded 635.0 g/kg, indicating moderate variations. The mustard straw substrates showed varying yields, ranged from 551.7 g/kg with wheat bran and 522.3 g/kg with molasses as compared to 482.4 g/kg without supplementation. The lowest yield was obtained on heat pasteurised sawdust (392.4 g/kg), which was not significantly better than all other treatments. Table 4.4: Effect of heat pasteurised substrates and their supplementation on growth characteristics and yield of P. djamor Yield* Pileus size* Stipe thickness* Treatment details (g/kg of dry weight of (cm) (cm) substrate) Wheat Straw 8.0 1.90 693.8 Wheat Straw + Wheat Bran 8.3 2.13 755.0 Wheat Straw + Molasses 8.1 1.93 736.4 Mustard Straw 6.7 1.60 482.4 Mustard Straw + Wheat Bran 6.9 1.68 551.7 Mustard Straw + Molasses 6.8 1.63 522.3 Paddy Straw 7.2 1.70 605.0 Paddy Straw + Wheat Bran 7.5 1.80 655.0 Paddy Straw + Molasses 7.4 1.73 635.0 Saw Dust 4.1 1.43 392.4 Saw Dust + Wheat Bran 4.3 1.57 440.0 Saw Dust + Molasses 4.2 1.48 421.7 CD at 5 % level 0.37 0.17 12.19 *Average of three replications and five bags per replication 28 29 4.3: Effect of chemically pasteurized substrates and their supplementation on growth characteristics as well as yield of P. pulmonarius 4.3.1 Effect of chemically pasteurized substrates and their supplementation on crop cycle of P. pulmonarius The results noticed in Table 4.5 revealed the data concerning the effect of chemically pasteurized substrates like wheat straw, mustard straw, paddy straw, saw dust and their supplementation i.e. wheat bran and molasses on crop cycle of P. pulmonarius. As per Table 4.5, the results on spawn run clearly showed that the time taken for spawn run of the different treatments varied from 14.3 days to 33.3 days. Among the different substrates, those enriched with wheat bran and molasses possessed shorter spawn run durations compared to their respective substrates without supplementation. It was observed that wheat straw supplemented with wheat bran exhibited significantly shortest period of 14.3 days for colonisation of substrates as compared to other treatments. It was followed by 15.7 days in wheat straw enriched with molasses. The paddy straw based treatments demonstrated spawn run periods varying from 16.7 days with wheat bran to 18.3 days with molasses and 20.7 days without supplementation, indicating moderate variations. While sawdust required significantly longest period of 33.3 days for completion of spawn run. According to the data revealed in Table 4.5, the duration of pinhead formation varied significantly among different treatments. The chemically pasteurised wheat straw supplemented with wheat bran had significantly shortest duration for pinhead formation at 18.3 days followed by wheat straw enriched with molasses at 18.7 days. The mustard straw based substrates showed longer periods for pinhead formation, ranged from 29.0 days with wheat bran to 33.0 days with molasses and 34.0 days without supplementation. While sawdust without supplements required the significantly longest at 40.7 days for pinhead emergence. Among the different treatments, days required for first flush of harvesting were significantly lowest in the wheat straw + wheat bran chemically pasteurised i.e. 24.3 days followed by wheat straw supplemented with molasses at 25.3 days. The paddy straw based treatments demonstrated first flush duration varied from 29.0 days with wheat bran to 32.3 days without supplementation and 29.3 days with molasses, indicating moderate variability. It was noticed that maximum number of 45.3 days were required for first harvest on sawdust which was pasteurised chemically. It was significantly longest period as compared to the other treatments. Data presented in Table 4.5 also revealed that the number of days required for the second harvest differed significantly among all the treatments. The second flush of P. pulmonarius was completed within 36.3 to 54.0 days in different treatments. It was significantly shortest at 36.3 days in wheat substrate supplemented with wheat bran. It was 30 followed by wheat substrate supplemented with molasses at 37.3 days. The wheat straw alone showed number of days required for the second harvest at 38.3 days. The mustard straw substrates showed longer durations for the second flush, ranged from 45.3 days with wheat bran to 50.3 days without supplementation and 48.7 days with molasses. While longest period of second harvest was observed in sawdust at 54.0 days which was significantly longest period as compared to the other treatments. The wheat straw supplemented with wheat bran exhibited significantly shortest crop duration of 48.3 days as compared to other treatments. It was followed by 49.3 days in wheat straw supplemented with molasses. The mustard straw substrates exhibited longer crop duration, with duration ranged from 56.3 days when supplemented with wheat bran to 60.0 days without supplementation and 59.7 days with molasses. The paddy straw treatments showed range from 50.7 days with wheat bran to 52.7 days without supplementation and 52.3 days with molasses. The sawdust required longest crop duration of 63.7 days and it was significantly longest crop duration as compared to the other treatments (Table 4.5). Table 4.5: Effect of chemical pasteurised substrates and their supplementation on crop cycle of P. pulmonarius Spawn Pinhead 2nd Crop 1st flush* Treatment details run* formation* flush* duration* (days) (days) (days) (days) (days) Wheat Straw 16.3 20.7 28.3 38.3 49.7 Wheat Straw + Wheat Bran 14.3 18.3 24.3 36.3 48.3 Wheat Straw + Molasses 15.7 18.7 25.3 37.3 49.3 Mustard Straw 27.7 34.0 41.3 50.3 60.0 Mustard Straw + Wheat Bran 23.7 29.0 36.3 45.3 56.3 Mustard Straw + Molasses 26.0 33.0 39.7 48.7 59.7 Paddy Straw 20.7 26.3 32.3 41.3 52.7 Paddy Straw + Wheat Bran 16.7 21.3 29.0 39.3 50.7 Paddy Straw + Molasses 18.3 23.0 29.3 41.0 52.3 Saw Dust 33.3 40.7 45.3 54.0 63.7 Saw Dust + Wheat Bran 29.3 36.7 42.3 52.3 61.3 Saw Dust + Molasses 31.7 38.3 43.7 53.7 62.3 CD at 5 % level 1.12 1.63 1.74 1.81 1.75 *Average of three replications and five bags per replication 31 4.3.2 Effect of chemical pasteurised substrates and their supplementation on growth characteristics and yield of P. pulmonarius The Table 4.6 presents the results from an experiment examining the effect of chemical pasteurised substrates and their supplementation on growth characteristics like pileus size (cm), stipe thickness (cm) and yield (g/kg of dry weight of substrate) of P. pulmonarius. The pileus size of fruit bodies of P. pulmonarius ranged from 4.7 to 9.9 cm among different treatments. The maximum pileus size of 9.9 cm was observed on chemically pasteurised wheat substrate enriched with wheat bran which was highly significant than other treatments, followed by pileus size of 9.7 cm in wheat straw enriched with molasses. The mustard straw substrates showed smaller pileus sizes, ranged from 7.2 cm without supplementation to 7.7 cm with wheat bran and 7.5 cm with molasses. The minimum pileus size of 4.7 cm was noticed in chemically pasteurised sawdust which was significantly minimum amidst all the treatments. Data provided in Table 4.6 revealed that among different treatments, highest mean stipe thickness of 2.40 cm was noticed on wheat substrate enriched with wheat bran. It was significantly more as compared to all other treatments. It was followed by wheat straw supplemented with molasses exhibiting a stipe thickness of 2.33 cm. The paddy straw treatments resulted in stipe thicknesses varied from 2.00 cm without supplementation to 2.13 cm with wheat bran and 2.07 cm with molasses. The mustard straw substrates showed thinner stipes ranged from 1.77 cm without supplementation to 2.03 cm with wheat bran and 1.87 cm with molasses. The minimum stipe thickness of 1.47 cm was observed in sawdust which was not found significantly better over all other treatments. The total yield ranged from 505.0 to 872.4 g per kg dry substrate on different treatments. The highest yield (872.4 g/kg dry substrate) was recorded on chemically pasteurised wheat straw supplemented with wheat bran which was significantly higher than all the other treatments (Table 4.6). It was followed by wheat straw enriched with molasses (862.4 g/kg dry substrate). The paddy straw also demonstrated good yields, ranged from 715.0 g/kg without supplementation to 770.0 g/kg with wheat bran and 753.4 g/kg with molasses. The mustard straw showed lower yields, ranged from 645.0 g/kg without supplementation to 684.7 g/kg with wheat bran and 670.3 g/kg with molasses. The lowest yield was obtained on chemically pasteurised sawdust (505.0 g/kg), which was not found significantly better over all other treatments. 32 Table 4.6: Effect of chemical pasteurised substrates and their supplementation on growth characteristics and yield of P. pulmonarius Yield* Pileus size* Stipe thickness* Treatment details (g/kg of dry weight of (cm) (cm) substrate) Wheat Straw 8.6 2.17 844.7 Wheat Straw + Wheat Bran 9.9 2.40 872.4 Wheat Straw + Molasses 9.7 2.33 862.4 Mustard Straw 7.2 1.77 645.0 Mustard Straw + Wheat Bran 7.7 2.03 684.7 Mustard Straw + Molasses 7.5 1.87 670.3 Paddy Straw 7.8 2.00 715.0 Paddy Straw + Wheat Bran 8.5 2.13 770.0 Paddy Straw + Molasses 8.3 2.07 753.4 Saw Dust 4.7 1.47 505.0 Saw Dust + Wheat Bran 5.1 1.67 548.3 Saw Dust + Molasses 4.9 1.57 605.0 CD at 5 % level 0.18 0.16 8.24 *Average of three replications and five bags per replication 4.4: Effect of heat pasteurised substrates and their supplementation on growth characteristics and yield of P. pulmonarius 4.4.1: Effect of heat pasteurised substrates and their supplementation on crop cycle of P. pulmonarius The results presented in Table 4.7 revealed the data concerning the effect of heat pasteurized substrates like wheat straw, mustard straw, paddy straw, saw dust and their supplementation i.e., wheat bran and molasses on crop cycle of P. pulmonarius. The results on spawn run clearly showed that the time taken for spawn run of the different treatments significantly varied from 20.3 days to 37.7 days. Among the different treatments, substrates supplemented with wheat bran and molasses exhibited shorter spawn run durations as compared to their respective substrates without supplementation. It was observed that wheat straw enriched with wheat bran exhibited significantly shortest period of 20.3 days for colonisation of substrates as compared to all the other treatments. It was followed by 21.7 days in wheat straw enriched with molasses. The wheat straw alone took 23.0 days for spawn run completion. The mustard straw based treatments exhibited longer spawn run durations with 32.0 days for mustard straw alone, 28.0 days with wheat bran and 29.7 days with molasses. The sawdust required significantly longest period of 37.7 days to complete spawn run. It is clear from Table 4.7 that the duration of pinhead formation varied significantly among different treatments between 24.3 to 44.0 days. The heat pasteurised wheat straw 33 enriched with wheat bran has significantly shortest duration for pinhead formation at 24.3 days followed by wheat straw enriched with molasses at 24.7 days. The wheat straw alone takes 28.0 days for pinhead formation to occur. The paddy straw based treatments ranged from 28.3 days with wheat bran to 32.3 days without supplementation and 30.3 days with molasses. The mustard straw treatments showed longer durations for pinhead formation with 36.0 days for mustard straw alone, 33.3 days with wheat bran and 35.3 days with molasses. Meanwhile, sawdust required significantly longest duration at 44.0 days for pinhead formation. Among the different treatments, days required for first flush of harvesting were significantly lowest in the wheat straw + wheat bran heat pasteurised i.e. 30.3 days, followed by wheat straw enriched with molasses at 31.3 days. The wheat straw alone required 35.7 days for the first flush. The paddy straw treatments ranged from 36.0 days with wheat bran to 38.3 days without supplementation and 36.7 days with molasses. The mustard straw treatments took longer duration for first flush of harvesting with 43.3 days for mustard straw alone, 40.7 days with wheat bran and 42.0 days with molasses. It was noticed that heat pasteurised sawdust required maximum number of days for first harvest at 48.7 days and it was significantly longest period as compared to the other treatments. Data presented in Table 4.7 revealed that among all the different treatments, the second flush of P. pulmonarius was completed within 40.3 to 55.7 days. It was significantly lowest at 40.3 days in wheat substrate supplemented with wheat bran and differed significantly among other treatments. It was followed by wheat substrate supplemented with molasses at 42.7 days. The paddy straw treatments ranged from 43.3 days with wheat bran to 45.3 days without supplementation and 43.7 days with molasses. The mustard straw treatments took longer durations ranged from 49.0 days when supplemented with wheat bran to 50.3 days for mustard straw without enrichment and 49.7 days with molasses. The sawdust required longest period of 55.7 days which was significantly longest period amidst all the treatments. The wheat straw supplemented with wheat bran exhibited significantly shortest crop duration of 50.7 days as compared to other treatments. It was followed by 51.3 days in wheat straw enriched with molasses. The wheat straw alone required 53.3 days for the crop duration. The paddy straw substrates showed crop durations of 54.0 days with wheat bran, 55.7 days with molasses and 57.0 days without supplementation. The mustard straw substrates exhibited longer crop durations i.e., 58.3 days with wheat bran and 59.0 days with molasses as compared to 59.3 days in mustard straw alone. The sawdust required longest crop duration of 63.7 days and it was significantly longest crop duration as compared to the other treatments (Table 4.7).