Agriculture and Natural Resources 50 (2016) 186e191 Contents lists available at ScienceDirect Agriculture and Natural Resources journal homepage: http://www.journals.elsevier.com/agriculture-and- natural-resources/ Original article Selection of Macrocybe crassa mushroom for commercial production * Tanapak Inyod,a, b Suriya Sassanarakit,b Achara Payapanon,c Suttipun Keawsomponga, a Department of Biotechnology, Faculty of Agro-Industry, Kasetsart University, Chatuchak, Bangkok 10900, Thailand b Thailand Institute of Scientific and Technological Research (TISTR), Technopolis, Klong 5, Klong Luang, Pathumthani 12120, Thailand c Department of Agriculture, 50 Phaholyothin Road, Chatuchak, Bangkok 10900, Thailand article info abstract Article history: Macrocybe crassa or Tricholoma crassum (Berk.) Sacc. (synonym) is a wild, edible mushroom in Thailand. Received 14 September 2015 This mushroom has a large fruiting body with meaty texture and a delicious taste. However, this Accepted 14 March 2016 mushroom is not commercially cultivated at a large scale. In this study, five strains of M. crassadDOA, Available online 25 June 2016 DOA-1, DOA-4, DOA-7 and DOA-10dwere cultivated on a ratio of rubber tree sawdust to fine rice bran to magnesium sulfate (MgSO4.7H2O) to calcium oxide (CaO) of 100:3:0.2:1 (weight per weight). Their Keywords: growth, productivity and characteristics of the fruiting body were investigated. The results revealed that Macrocybe crassa DOA-10 gave the highest yield (215.10 g per 0.6 kg of substrate) with 59.26% biological efficiency. Nutritional value Productivity Observation showed that its pileus diameter, stalk diameter and stalk length were 6.78 cm, 2.75 cm and Quality 13.54 cm, respectively. The dried samples of DOA-10 contained protein, carbohydrates, ash and fat at Substrate 13.71%, 68.08%, 12.06% and 2.49% respectively. Moreover, they contained high contents of both macro- nutrients and micronutrients: potassium (43,100 mg/kg), calcium (492.00 mg/kg), magnesium (1046.66 mg/kg), sodium (936.33 mg/kg), iron (283.21 mg/kg), zinc (53.79 mg/kg) and manganese (17.55 mg/kg). These results confirmed that DOA-10 is a promising strain for commercial cultivation. Copyright © 2016, Kasetsart University. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/). Introduction They produce a fleshy convex pileus and can be found in every region of Thailand and in neighboring countries (Payapanon and Mushroom cultivation is a major agro-industry and it has Srijumpa, 2008). Several related species such as Tricholoma albo- become popular as a source of healthy and functional food, and its brunneum, Tricholoma flavovirens, Tricholoma paneolum, Tricholoma production is also increasing (Aida et al., 2009). Macrocybe crassa or equestre, Tricholoma terreum and Tricholoma georgii, are found in Tricholoma crassum (Berk.) Sacc. (its synonym) is a wild, edible Europe and America while only a few edible species of this family, mushroom and non-toxic (Teaumroong et al., 2002; Pradhan et al., such as Tricholoma matsutake and Macrocybe crassum, are found in 2010). It is a fungus belonging to the genus Macrocybe, in the family the Asian regiondin Japan, Thailand and Sri-Lankad(Huffman of Tricholomataceae, and the order of Agaricales (Chang and Hayes, et al., 1989). In Thailand, M. crassum has various common names 1978). It has a high production yield with outstanding, enormous, among local people such as hed-tin-rad (northeast), hed-jan (north), fleshy basidiomata which may exceed 80 kg fresh weight and it hed-hua-sum (south) and hed yai or hed-tub-tao-khao (central) shows high biological efficiency, which is more than for many (Petcharat, 1996). Natural M. crassa is generally rather expensive edible mushrooms (Corner, 1993). The large size and flavorful and rare, because it is usually found only once a year, particularly in fruiting bodies of some Macrocybe species make them a valuable the rainy season (Teaumroong et al., 2002). Moreover, varying the source of food, such as lower cholesterol and immuno-modulating substrate media for mushroom cultivation resulted in different effects in humans (Hoshi et al., 2005). Species of Macrocybe were yields due to biological and chemical differences between the formerly placed within Tricholoma, but segregated in the section substrate media and their quality which may been due to the ge- Leucorigida morphologically on the presence of clamp connections, notype of the mushroom strains (Ragunathan and Swaminathan, and ecologically on the absence of ectomycorrhizal associations 2003). However, it can easily and successfully be cultivated on (Pegler et al., 1998). rubber tree sawdust (Payapanon and Srijumpa, 2008). However, still no work has been done to determine the suitability of this locally available lignocellulosic waste for cultivation or to deter- * Corresponding author. mine the most cost compatible strains under environmental E-mail address: [email protected] (S. Keawsompong). http://dx.doi.org/10.1016/j.anres.2016.06.006 2452-316X/Copyright © 2016, Kasetsart University. Production and hosting by Elsevier B.V. This is an open access article under the CC BY-NC-ND license (http:// creativecommons.org/licenses/by-nc-nd/4.0/). T. Inyod et al. / Agriculture and Natural Resources 50 (2016) 186e191 187 conditions. The main objectives of this research were to evaluate rehydrated basidiocarps using a sharp surgical blade. The sections the yield parameters of different strains of M. crassa through were immersed in a diluted solution of methyl blue stain and traditional cultivation methods on rubber tree sawdust substrates covered with cover slips. The color and size of basidiospores were and to determine their growth and the productivity and charac- examined. teristics of the fruiting body to identify the best strain of M. crassa mushroom most suitable for commercial cultivation. Nutritional analysis Materials and methods The protein, fat, ash, crude fiber and total carbohydrate contents were determined using the procedures recommended by Mycelial development of Macrocybe crassa mushroom Association of Official Analytical Chemists (1995) and Manzi et al. (2001). The mineral contents of M. crassa mushroomsdpotassium fi Mycelial discs (10 mm diameter) of ve native strains (with (K), calcium (Ca), magnesium (Mg), sodium (Na), iron (Fe), zinc d substantially high production yield) DOA, DOA-1 (collected from (Zn), copper (Cu)dand the manganese (Mn), lead (Pb), cadmium Bangkok province), DOA-4 (collected from Chiang Rai province), (Cd) and arsenic (As) contents were determined using atomic ab- DOA-7 (collected from Chai Nat province) and DOA-10 (collected sorption spectrophotometry (Willard et al., 1988). Total phosphorus d from Prachuap Khiri Khan province) were obtained from the was measured using the phosphorus-vanodomolybdate method strain bank of the Department of Agriculture, Ministry of Agricul- (Munson and Nelson, 1990). The glucan concentration was ture and Cooperative, Bangkok, Thailand, and were carefully measured using a Mushroom and Yeast Glucan Assay Kit (K-YBGL transferred to the upper surfaces of prepared sorghum grains. 09/2009; Megazyme International Ireland Ltd.; Bray, County Inoculated grain bottles were tightly secured using moist cotton Wicklow, Ireland). wool and covered with sterile aluminum foil. They were kept in dark, sterile cabinets at ambient room temperature for 14 d until Statistical analysis they were fully colonized. The substrates employed in the pro- duction and in-depth mycelial development assessments of Mac- A completely randomized design was used in all experiments. rocybe were prepared using rubber tree (Hevea brasiliensis) sawdust Data collected were subjected to ANOVA at the 5% level of signifi- mixed with fine rice bran, magnesium sulfate (MgSO4.7H2O) and cance using the SAS version 9.1 software package (SAS Institute, calcium oxide (CaO) in the ratio of 100:3:0.2:1 (weight per weight). 2005). Water was added to adjust the moisture content to 65%. Samples of the substrate mixture (each approximately 600 g) were used to fill 1 kg capacity autoclavable polypropylene bags to only three- Results and discussion quarters of their capacity and then steamed for 3 h in a large cast-iron steamer. When the temperature in the mushroom spawn Mycelial development of Macrocybe crassa mushroom cooled down to room temperature, 10e15 seeds of grains spawn were added to the bags and incubated at approximately 25 C for The findings showed that mycelial colonies of all strains of spawn running until the bags were fully covered with mycelia. Data M. crassa were white and cottony, with abundant aerial hyphae. The were collected on a daily basis for the mycelium running rate mycelia running through the substrate could be observed 3e4d (MRR), days required to complete spawn running (DCS), primordial after spawn addition, and the isolates slowly progressed down the initiation (DFPI) and the number of fruiting bodies (NFB). formulations completely ramifying them at different rates. The daily mycelial growth rates of different strains are presented in Production on sterile substrate Table 1. The mushroom strain DOA-10 had the fastest complete spawn running of 35 d on the sawdust substrate whereas DOA, The upper end of the bags was cut off and the spawn mush- DOA-4 and DOA-7 took about 37 d for full saturation. Yadav et al. rooms were neatly arranged in a basket and placed on shelves in (2013) reported that the spawn running of Tricholoma sp. in poly- the growing room. The surface of the compost was covered with a thene bags was completed in 30e35 d. The highest MRR was 2.5e5 cm thick layer of steamed casing soil, which was necessary to observed for DOA-10 (0.42 cm/d) which was significantly different initiate fruiting. After casing, the relative humidity of the room was from the MRR of DOA-7 (0.39 cm/d), DOA-1 (0.39 cm/d), DOA-4 maintained between 85% and 90%.