Micología Aplicada International ISSN: Colegio de Postgraduados México

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1 Micología Aplicada International ISSN: Colegio de Postgraduados México Bernabé-González, T.; Cayetano-Catarino, M. Cultivation of Pleurotus pulmonarius on substrates treated by immersion in alkaline water in Guerrero, Mexico Micología Aplicada International, vol. 21, núm. 1, enero, 2009, pp Colegio de Postgraduados Puebla, México Available in: How to cite Complete issue More information about this article Journal's homepage in redalyc.org Scientific Information System Network of Scientific Journals from Latin America, the Caribbean, Spain and Portugal Non-profit academic project, developed under the open access initiative

2 Mi c o l o g i a Ap l i c a d a In Cultivation t e r n i o n a l, of 21(1), Pl e u r2009, o t u s pp , Be r k e l e y, CA, U.S.A. Cu lt i vat i o n o f Pl e u r o t u s p u l m o n a r i u s o n s u b s t r at e s t r e at e d b y i m m e r s i o n in alkaline water in Guerrero, Mexico T. Be r n a b é-go n z á l e z a n d M. Cay e ta n o-cata r i n o Academic Unit of Biological and Chemical Sciences, Autonomous University of Guerrero, Av. Lázaro Cárdenas s/n, Chilpancingo 39090, Guerrero, Mexico. teobernaglez@hotmail.com Accepted for publication August 15, 2008 ABSTRACT Pleurotus pulmonarius was cultivated on dry banana leaves (Musa paradisiaca) or dry palmareca leaves (Chrysalidocarpus lutescens), using two substrate treatments. Substrates were immersed in water containing 2% lime for 24 h and used for mushroom cultivation, in comparison with the method of immersing substrates in hot water at ±80 C for 1 h. The highest mushroom production was obtained on banana leaves immersed in alkaline water, reaching 120.1% of biological efficiency in a period of 61 days. Biological efficiency in the other treatments varied between %. Substrates and treatments studied are suitable for low-cost and small-scale production of oyster mushrooms. Key words: Agricultural by-products, mushroom cultivation, Pleurotus pulmonarius, alkaline substrate treatment. INTRODUCTION Oyster mushrooms (Pleurotus spp.) are distributed worldwide, and several species are grown commercially on a large and small scale in many countries. As primary decomposers capable of degrading lignocellulose, they are cultivated on a variety of treated and even untreated agricultural by-products 2. Diverse methods of substrate treatment have been used involving the use of chemicals (e.g.,

3 20 T. Be r n a b é-go n z á l e z a n d M. Cay e ta n o-cata r i n o fungicides, formaldehyde, lime, hydrogen peroxide, biodegradable detergents), fermentation processes, hot water, and steam 9,11. The immersion of substrates in hot water was developed by Kurtzman in Later, he refined it 8. Variations of this treatment (±80 C, 1 h) are common in the cultivation of oyster mushrooms in Mexico 1,4,5. Alternative rustic procedures have also been tested. Villa Cruz et al. 13 studied a mixture of coffee pulp and corn cobs, treated only with a water solution of lime (2%) or fungicide (100 ppm benomyl), as substrate for the cultivation of P. ostreatus. They recorded biological efficiencies (BEs) of %, and found no significant differences in relation with those from the same substrate fermented for several days. Hernández et al. 6 cultivated P. ostreatus on a mixture of grass (70%) and coffee pulp (30%), pre-soaked in a water solution of 2% lime and fermented (2-3 days), reaching BEs of 59-93%. Contreras et al. 3 reported BEs ranging from % for P. ostreatus, following the same procedure but using a water solution of 0.5% lime. In this study, we cultivated P. pulmonarius using dry banana leaves and on dry palmareca leaves, a common ornamental palm, as substrates. Mushroom yield, BE, and production rate were recorded on these substrates treated with a water solution of 2% lime for one day, and compared with the method of treating substrates with hot water (±80 C) for one hour. MATERIALS AND METHODS Strain. Pleurotus pulmonarius (Fr.) Quél. (strain IE-4) was used in all experiments. This strain is deposited at the culture collection of the Institute of Ecology (Xalapa, Mexico). It was maintained and subcultured on potato dextrose agar (PDA, Bioxon). Spawn preparation. This was carried out according to a method previously described 11. Sorghum grains (Sorghum vulgare Pers.) were soaked in water for 24 h, drained, autoclaved (121 C for 1 h) in polypropylene bags (ca. 150 g/bag), cooled, and inoculated with P. pulmonarius. Inoculated bags were incubated at 29 C for two weeks in darkness. Substrates. Dry banana leaves (Musa paradisiaca L.) were obtained from the banana-producing region at Tecoanapa, Guerrero, Mexico. Dry palmareca leaves (Chrysalidocarpus lutescens H. Wendl.) were collected from public gardens of Chilpancingo city, Guerrero. These substrates were fragmented (3-5 cm) manually using a machete. Treatment with lime. Substrates were submerged separately in a tap water solution of 2% lime [Ca(OH) 2 ] for 24 h using plastic trays. Treatments were kept at ambient temperature, including banana leaves (BTL) and palmareca leaves (PTL) treated with lime. BTL and PTL had the same ph of 9.0, and a moisture content of 76.9% and 63.0%, respectively. Treatment with hot water. Substrates were wetted separately by immersion in tap water for 24 h. Thereafter, they were treated by immersion in hot water at 80 C for 1 h. Treatments were banana leaves (BTH) and palmareca leaves (PTH) treated with hot water. BTH and PTH had the same ph of 6.0, and a moisture content of 78.8% and 65.5%, respectively. Mushroom cultivation. Ten samples (ca. 3 kg, fresh weight) of each substrate treatment were placed into plastic bags (50 x 70 cm), and inoculated with 150 g of spawn (5%, w/w). All inoculated bags were incubated

4 Cultivation of Pl e u r o t u s 21 at 26±3 C in darkness until first primordia appeared. In the fruiting room, there were daylight conditions through windows, and the minimum temperature was 24 C, the maximum temperature was 29 C, and the relative humidity was 75-80%. Variables studied were: the number of days until primordia formation, mature mushroom yield during three flushes (fresh weight of mushrooms/fresh weight substrate x 100), biological efficiency (BE, fresh weight of mushrooms/dry weight substrate x 100), and production rate (PR, BE/total days of production) 1,2. Experimental design and statistical analysis. Data were processed by analysis of variance in a complete randomized bifactorial design. Significant differences between means were analyzed by the Tukey s multiple range test (α = 0.05%). RESULTS AND DISCUSSION Musa leaves and Chrysalidocarpus leaves were suitable substrates for mycelial growth and fruiting of P. pulmonarius (Fig. 1). There was no contamination by moulds in all samples studied, although several samples treated with lime showed some bacterial contamination at the end of experiments. First primordia appeared earlier on Chrysalidocarpus leaves treated with hot water (PTH, 28 days), and Musa leaves treated with lime (BTL, 30 days). On Musa leaves treated with hot water (BTH) and Chrysalidocarpus leaves treated with lime (PTL), primordia appeared 36 and 37 days after spawning (Table 1). The highest total mushroom production was obtained on BTL (832.5 g), followed by BTH (516.7 g), reaching a biological efficiency (BE) of 120.1% and 81.2%, respectively (Table 2). In these cases, the mushroom yield was 27.7% (BTL) and 17.2% (BTH), while the production rate (PR) was 2.0% (BTL) and 1.2% (BTH). Their total production cycle was similar, being of 61 days for BTL and 63 days for BTH. PTL and PTH treatments showed lower total mushroom production ( g), BE ( %), yield ( %), and PR ( %). The BE obtained on BTL (120.13%) with P. Table 1. Period after spawning for primordia formation of Pleurotus pulmonarius on substrates studied. Primordia appeared (days after spawning) Treatment 1st 2nd 3rd Total days of production cycle Musa leaves treated with lime 30±5.6 b 1 42±5.6 b 57±6.2 b 61±6.2 b Musa leaves treated with hot water (±80 C, 1 h) 36±4.2 a 48±3.1 ab 59±5.0 ab 63±5.0 ab Chrysalidocarpus leaves treated with lime 37±3.1 a 50±3.4 a 64±3.1 a 68±3.3 a Chrysalidocarpus leaves treated hot water 28±4.6 b 42±6.2 b 54±7.6 b 58±7.5 b 1 Different letters in the same column indicate significant differences according to Tukey s multiple range test (α = 0.05).

5 22 T. Be r n a b é-go n z á l e z a n d M. Cay e ta n o-cata r i n o Table 2. Average yield of fresh mushrooms from Pleurotus pulmonarius cultivated on substrates studied. Harvest (g) Treatment DWS 1st 2nd 3rd Total Biological Yield Production (kg) efficiency (%) rate (%) (%) Musa leaves treated with lime ± a 1 310±43.81 a 172±31.87 a 832.5±99.90 a ±14.54 a 27.75±3.36 a 2.03±0.32 a Musa leaves treated with hot water (±80 C, 1 h) ±30.80 b 177.5±42.24 b 158.5±30.91 a 516.7±54.52 b 81.24±8.57 b 17.22±1.72 b 1.29±0.17 b Chrysalidocarpus leaves treated with lime ±29.87 b 187.4±43.24 b 73.5±25.93 b 460.2±49.05 b 41.46±4.41 c 15.34±1.64 b 0.61±0.07 c Chrysalidocarpus leaves treated with hot water ±61.19 b 143.9±72.36 b 82.4±37.32 b 465.2±81.41 b 44.95±7.86 c 15.51±2.57 b 0.78± 0.18 c DWS= Dry weight substrate. 1 Different letters in the same column indicate significant differences according to Tukey s multiple range test (α = 0.05). pulmonarius is similar to that recorded on other substrates, such as fermented tequila maguey bagasse (84.0%) 10 and fermented sugar cane bagasse (51.0%) 4. However, it was lower than the BE recorded on the coffee pulp (138%) 12. Overall analysis showed that the BTL treatment was significantly different from the rest of the treatments BTH, PTL, PTH. In fact, leaves from Chrysalidocarpus showed poor water Fig. 1. Old fruit bodies of Pleurotus pulmonarius cultivated on Musa leaves treated with lime. retention capacity, and accordingly low moisture content (63.0%). Substrates and treatments studied are suitable for lowcost and small-scale production of oyster mushrooms. LITERATURE CITED 1. Bernabé-González, T., M. Cayetano-Catarino, A. Adán-Díaz and M. A. Torres-Pastrana Cultivo de Pleurotus pulmonarius sobre diversos subproductos agrícolas de Guerrero, México. Revista Mexicana de Micología 18: Chang, S. T. and P. G. Miles Edible Mushrooms and Their Cultivation. CRC. Press, Boca Raton. 345 pp. 3. Contreras, E. P., M. Sokolov, G. Mejía and J. E. Sánchez Soaking of substrate in alkaline water as a pretreatment for the cultivation of Pleurotus ostreatus. Journal of Horticultural Science & Biotechnology 79: Guzmán-Dávalos, L., C. Soto and D. Martínez- Carrera El bagazo de caña de azúcar como substrato para la producción de Pleurotus en Jalisco. Revista Mexicana de Micología 3: Guzmán-Dávalos, L., D. Martínez-Carrera, P. Morales and C. Soto El cultivo de hongos comestibles (Pleurotus) sobre el bagazo del maguey de la industria tequilera. Revista Mexicana de Micología 3:

6 Cultivation of Pl e u r o t u s Hernández, D., J. E. Sánchez and K. Yamasaki A simple procedure for preparing substrate for Pleurotus ostreatus cultivation. Bioresource Technology 90: Kurtzman, R. H., Jr From straw to supper in five days. Pp In: Proceedings of Seminar on Mushroom Research and Production. Ed. I. Husain. Agricultural Research Council, Karachi, Pakistan. 8. Kurtzman, R. H., Jr Production and cultivation of mushrooms in the west, particularly Europe and North America. Food Reviews International 13: Oei, P Mushroom Cultivation III. Backhuys Publishers, Leiden. 429 pp. 10. Soto-Velazco, C., L. Guzmán-Dávalos and L. Villaseñor Substrates for cultivation of Pleurotus in Mexico, I. Tequila maguey bagasse (Agave tequilana). Mushroom Journal for the Tropics 11: Stamets, P Growing Gourmet and Medicinal Mushrooms. Ten Speed Press, Berkeley, California. 552 pp. 12. Velázquez-Cedeño, M. A., G. Mata and J.-M. Savoie Waste-reducing cultivation of Pleurotus ostreatus and Pleurotus pulmonarius on coffee pulp: changes in the production of some lignocellulolytic enzymes. World Journal of Microbiology & Biotechnology 18: Villa-Cruz, V., G. Huerta-Palacios and J. E. Sánchez-Vázquez Fermentation of a mixture of corn-cobs and coffee pulp for the cultivation of Pleurotus ostreatus. Micología Neotropical Aplicada 12: