PRODUCTION of Panus Strigellus SPAWN USING the INTERNAL SHEATH of PEACH PALM (Bactris Gasipaes) AS a SUBSTRATE

PRODUCTION OF Panus strigellus SPAWN USING THE INTERNAL SHEATH OF PEACH PALM (Bactris gasipaes) AS A SUBSTRATE

Ruby Vargas-Isla, Lucia Kiyoko Ozaki Yuyama, Jaime Paiva Lopes Aguiar and Noemia Kazue Ishikawa

SUMMARY

Mycelial growth takes place at elevated temperatures in Pa- analysis in Petri dishes of the effects of substrate moisture and nus strigellus, which makes it a promising species for cultiva- sawdust supplementation levels. Mycelial growth was evaluated tion in tropical regions. The use of locally available substrates by measuring the growth (cm/day) and vigor of the colony. A is the first step in cost-effective mushroom production. Peach humidity of 60% and supplementation of Simarouba amara saw- palm, Bactris gasipaes is grown in palm agribusinesses in Ama- dust with PPIS in a 10:1 ratio was considered the most suitable zonas State, Brazil; one of the waste products of this crop is combination. This formulation was applied for spawn prepara- its protective internal sheath (PPIS). The potential use of PPIS tion in polypropylene bags. Spawn production of P. st r igellus for P. st r igellus spawn production was evaluated through the was successfully achieved after 25 days of incubation at 35°C.

USO DE LA VAINA INTERNA DEL PIJUAYO (Bactris gasipaes) PARA LA PRODUCCIÓN DE SEMILLA-INÓCULO DE Panus strigellus Ruby Vargas-Isla, Lucia Kiyoko Ozaki Yuyama, Jaime Paiva Lopes Aguiar y Noemia Kazue Ishikawa RESUMEN

Panus strigellus presenta crecimiento micelial en temperatu- ducción de semilla-inóculo de P. st r igellus, analizando el efecto ras elevadas, favoreciendo el desarrollo de su cultivo en regio- de la humedad del substrato y los niveles de suplementación de nes tropicales. Obtener substratos localmente disponibles es el aserrín por PPIS. El crecimiento micelial fue evaluado por la primer paso para el cultivo rentable de setas. El pijuayo, Bac- medida de la colonia en cm/día y el vigor. La humedad de 60% tris gasipaes, es cultivado en las agroindustrias de palmito en y Simarouba amara suplementada con PPIS en la proporción el Estado de Amazonas, Brasil; uno de los residuos originados 10:1 fue considerado como el mejor resultado. Esta formulación de la palmera es la vaina interna protectora (PPIS). Se evaluó fue aplicada en la elaboración de la semilla-inóculo de P. st r i- en placas de Petri el potencial de uso de la PPIS para la pro- gellus con éxito después de 25 días de incubación a 35°C.

Introduction ‘peach palm’, known locally as subsistence products, and cess, generating large amounts ‘pupunha’, as ‘pijuayo’ in Peru, heart of palm production is an of unused residue. Some stud- In Brazil, heart of palm is ‘pijiguao’ in Venezuela, expanding agribusiness (Silva ies have examined the use of extracted from several genera ‘pejibaye’ in Costa Rica and and Clement, 2005). The mi- these residues in mushroom and species of palm. Although Nicaragua, ‘chontaduro’ in Co- cro-businesses of peach palm cultivation. Tonini et al. Euterpe edulis Mart. (‘juçara’) lombia and Ecuador, ‘tembé’ in in Amazonas State, including (2007) investigated the use of and E. oleracea Mart. (‘açaí’) Bolivia, and ‘pibá’ in Panama the Manaus region, produced E. edulis sheaths as a medium have been used for heart of (http://en.wikipedia.org/Bac- 45.1ton of heart of palm in for the cultivation of Lentinula palm production, more heart of tris_gasipaes). 2012 (IDAM, 2012). The pro- edodes (Berk.) Pegler in ax- palm has been produced Bactris gasipaes is the only tective internal sheath of B. enic culture and obtained suc- (Clement and Bovi, 2000) us- domesticated Neotropical palm gasipaes is discarded during cessful basidiomata produc- ing Bactris gasipaes Kunth or whose starchy-oily fruits are the palm core production pro- tion. Sales-Campos and An-

KEYWORDS / Bactris gasipaes / Lentinus strigellus / Mushroom production / Panus strigellus / Simarouba amara / Substrates / Received: 11/13/2013. Modified: 10/30/2013. Accepted: 11/25/2013.

Ruby Vargas-Isla. Agronomical Lucia Kiyoko Ozaki Yuyama. Jaime Paiva Lopes Aguiar. Wood Noemia Kazue Ishikawa. Biolo- Engineer, Universidad Nacional Nutritionist, Universidade Fed- Industry Operational Engineer, gist, Universidade Estadual de de la Amazonía Peruana eral Fluminense, Brazil. Máster Universidade de Tecnologia do Londrina, Brazil. Master in Ag- (UNAP), Peru. M.Sc. in Humid in Food Sciences, Universidade Amazonas, Brazil. Especialist in ricultural Microbiology, Univer- Tropics Agriculture and Doctor Federal do Amazonas, Brazil. Nutrition and Food Sciences, sidade Federal de Viçosa, Bra- in Botany, Instituto Nacional de Doctor in Food Sciences, Uni- Instituto de Nutrición de Centro zil. Doctor in Natural Resourc- Pesquisas da Amazônia (INPA), versidade de São Paulo, Brazil. América y Panamá, Guatemala. es, Hokkaido University, Japan. Brazil. e-mail: rubyvar9@gmail. Researcher, INPA, Brazil. Researcher, INPA, Brazil. Researcher, INPA, Brazil. com

OCT 2013, VOL. 38 Nº 10 0378-1844/13/10/733-04 $ 3.00/0 733 USO DA BAINHA INTERNA DA PUPUNHEIRA (Bactris gasipaes) PARA PRODUÇÃO DE SEMENTE-INÓCULO DE Panus strigellus Ruby Vargas-Isla, Lucia Kiyoko Ozaki Yuyama, Jaime Paiva Lopes Aguiar e Noemia Kazue Ishikawa RESUMO

Panus strigellus apresenta crescimento micelial em tempera- llus. Em placas de Petri foram analisados o efeito da umidade turas elevadas, favorecendo o desenvolvimento do cultivo em do substrato e níveis de suplementação de serragens por PPIS. O regiões tropicais. Obter substratos disponíveis localmente é o crescimento micelial foi avaliado pela medida da colônia em cm/ primeiro passo para cultivo rentável de cogumelos. A pupunha, dia e vigor. A umidade de 60% e Simarouba amara suplementada Bactris gasipaes, é cultivado nas agroindústrias de palmito no com PPIS na proporção 10:1 foi considerada o melhor resultado. Amazonas, Brasil; um dos resíduos da palmeira é sua bainha in- Esta formulação foi aplicada na elaboração da semente-inóculo terna protetora (PPIS). Neste trabalho foi avaliado o potencial de em sacos de polipropileno. Obteve-se com sucesso a semente-inó- uso do PPIS para a produção de semente-inóculo de P. st r ige- culo de P. st r igellus após 25 dias de incubação a 35°C.

drade (2010) studied Panus dust spawn are inoculated with on Campus III, INPA, to ob- tion (Vargas-Isla et al., 2012). lecomtei (Fr.) Corner (Lentinus grain spawn into the sawdust tain PPIS. The samples were Incubation temperature was strigosus Fr.) mycelia growth formulation. 3) The substrate is weighed and divided into stipe, 35°C. on B. gasipaes stipes. formulated, usually using a car- leaves, external and internal Mycelial growth was evalu- Panus strigellus (Berk.) bon source such as sawdust, sheath, and heart of palm. Fol- ated by measurement colony Overh. has very interesting sugar cane (Saccharum offici- lowing dehydration at 65°C diameter change (cm/day) and prospect for the development of narum L.) bagasse, or corn (Zea during 48h, PPIS was crushed colony vigor was visually eval- mushroom cultivation in the mays L.) cobs, supplemented in an industrial blender (LSP- uated and classified as (+) thin, Amazonas State, considering its with a nitrogen source such as 04, Siemsen Ltd., Brusque-SC, (++) medium or (+++) dense. characteristics of mycelial cereal bran. 4) Spawn are in- Brazil) and then transferred growth at 35-40°C and accentu- oculated into the substrate of and ground in a Willye TE- Substrate formulations ated umami taste (Vargas-Isla mushroom culture. 5) Spawn 680 (Tecnal, Piracicaba-SP, and Ishikawa, 2008). Screening running. 6) Fruiting induction, Brazil) grinder, using a The experiment with differ- has been conducted using 11 e.g. by thermal shock, mechani- 0.5mm-mesh sieve. Sawdust of ent substrate formulations was forest species and regional fruit cal shock, injury and illumina- S. amara and Hymenolobium carried out in two stages. First, residues of the Central Amazon tion. 7) Fructification. 8) Har- petraeum Ducke (‘angelim-pe- the PPIS and S. amara sawdust to produce the spawn of P. stri- vesting. dra’), A. aculeatum fruit shell were mixed and supplemented gellus; among these, Simarouba The spawn constitutes the (TFS), and rice bran (Oryza separately with TFS and rice amara Aubl. (‘marupá’) sawdust base for the commercial culti- sativa L.) were used for com- bran in proportions of 10:1 and and Astrocaryum aculeatum vation of edible mushrooms, parison. Each residue was ov- 5:1 (w/w), and mixed with dis- Meyer (‘tucumã’) fruit shell and spawn production is the en-dried at 65°C with air cir- tilled water to ~60% hydration formulations presented the best main challenge faced by com- culation and stored in plastic (w/v). Pure S. amara sawdust alternatives for P. strigellus mercial mushroom producers. bags at room temperature was used as a control. Second, spawn production (Vargas-Isla The current study evaluated (~25°C). sawdust of H. petraeum and S. et al., 2012). However, the fruit formulations using the B. gasi- amara was prepared and mixed shell residues are only produced paes internal sheath (PPIS) as Centesimal analysis separately with PPIS and rice in small amounts by fruit pro- an alternative for use in a sub- bran (1:1 and 10:1 w/w), with cessing facilities and are thus strate for P. strigellus spawns. The centesimal composition distilled water added to ~60% unlikely to be available on the analysis of the residues was hydration (w/v). The formula- scale required for spawn pro- Material and Methods conducted in accordance with tions were then distributed on duction. the AOAC (1998) methodology Petri plates (15 ±1 g/plate) and The main steps for mush- Microrganism (n= 3). sterilized. The experiment was room production are eight, as evaluated after five days using follows. 1) Stock culture is first The P. strigellus INPACM Substrate humidity the criteria of substrate humid- grown on sterilised media in 1464 culture from the Coleção ity test of mycelial growth Petri plates and/or in tubes. 2) de Microrganismos de Interesse For the humidity measure- (growth measurement in cm/ Spawn or inocula are prepared, Agrossilvicultural, Instituto ment, sawdust of H. petraeum day and visual determination where the spawn are any form Nacional de Pesquisas da mixed with rice bran of colony vigor). of mycelia that can be dispersed Amazônia (INPA), was used. (sawdust:supplement 10:1 w/w) into a substrate. The most com- The stock culture and inocula was used and distilled water Statistical analysis mon forms of spawn substrate for the experiments were the was added to the formulation are grain or sawdust; cereal same as those reported by Var- to reach 30, 40, 50, 60 and Each experiment was con- grains such as wheat (Triticum gas-Isla et al. (2012). 70% hydration (w/v) prior to ducted twice using five repli- aestivum (L.) Thell.) may be distribution into Petri plates cates. Analysis of variance also used for spawn production. Residues for substrate (15 ±1 g/plate) and sterilized (ANOVA) was used to examine The next step is the exponential formulation 1h at 121°C. Following sterili- the results of the experiments, expansion of the mycelial mass. zation, one mycelial disk was and the averages were com- The pure culture is transferred Bactris gasipaes stipes deposited in the center of the pared using the Tukey test at to sterilised grain. Then, saw- (1.50m height; n= 5) were cut plate containing the formula- the 1% level of significance.

734 OCT 2013, VOL. 38 Nº 10 TABLE I In Brazil, the base material in DISTRIBUTION OF FRESH MATERIAL OBTAINED FROM Bactris gasipaes * the major substrates used for ed- Parts of palm ible mushroom cultivation in the south and southeast include rice Stipe Leaves External sheath Internal sheath Heart of palm straw and sugar cane bagasse for Fresh weight (kg) 17.9 5.5 1.7 2.4 0.7 Agaricus bisporus (J.E. Lange) Percentage 63.5 19.5 6.0 8.5 2.5 Pilát; Eucalyptus sp. and Quercus acutissima Carr. for L. *Data shown represent the average of five samples of ~1.5m height. edodes; and sugar cane bagasse (Saccharum officinarum L.) for TABLE II Pleurotus spp. (Bononi and COMPOSITION (% DRY MATTER) OF THE RESIDUES OBTAINED Trufem, 1986) IN MANAUS, AM, BRAZIL * Alternative sources have also Residues Ash Protein Lipids been evaluated, such as corn cob Astrocaryum aculeatum fruit shell 3.28 ±0.049 5.55 ±0.041 24.29 ±0.081 (Eira et al., 2005), elephant Bactris gasipaes internal sheath 3.79 ±0.040 3.65 ±0.046 0.88 ±0.062 grass (Pennisetum purpureum Oryza sativa bran 3.28 ±0.407 13.07 ±0.701 14.76 ±0.002 Schum.; Donini et al., 2005; Simarouba amara sawdust 0.13 ±0.012 1.34 ±0.034 0.42 ±0.0003 Bernardi et al., 2007), coffee husks (Coffea arabica L.; Fan et *Data shown represent the average of three samples according to AOAC (1998) methods. al., 2006), peach palm stipe (B. gasipaes; Sales-Campos and Spawn production were taken out of the packing fresh mushroom marketing in Andrade, 2010), pineapple crown and cut for visual observation the Manaus region is based on (Ananas comosus (L.) Merr.), S. amara sawdust supple- (n= 5). two species: shiitake (L. ‘cupuaçú’ fruit shell (Theobroma mented with PPIS (10:1) was edodes) and shimeji (Pleurotus grandiflorum (Willd. ex Spreng.) placed in polypropylene pack- Results and Discussion spp.) imported from Sao Paulo K. Schum.), banana peel (Musa ing (23×36cm) with 800g of State. sp.), and ‘tucumã’ fruit shell wet substrate with a respira- Table I lists the quantity Supplementation with a nitro- (Aguiar et al., 2011). tor, using a ring of PVC tub- and distribution of heart of gen source is necessary because Considering both criteria of ing with 3cm height × 5cm palm and residue obtained pure S. amara sawdust contains mycelial growth, in the present diameter and hydrophobic from B. gasipaes. The total only 1.34% crude protein (Table case the best substrate condition cotton for gas exchange (Var- residue is equivalent to 97.5%, II). The PPIS contained more was provided by 60 and 70% gas-Isla et al., 2012). The of which the internal sheath protein (3.65%) than S. amara humidity (p<0.01; Figure 1). The substrate package was steri- residue represents 8.5%. Con- sawdust but less protein than best substrate moisture depends lized for 1h at 121°C. Follow- sidering these data and the rice bran (13.07%). The nutri- on the mushroom cultivation spe- ing sterilization, one mycelial 2012 production of heart of tional supplements were added cies; e.g. 55-70% substrate mois- disk was deposited in the palm from B. gasipaes in the to increase the levels of nitrogen ture for L. edodes and 60-80% center of the plate containing Amazonas State, we estimated and useable carbohydrates be- substrate moisture for Pleurotus the formulation (Vargas-Isla that over 1768ton of residue cause nitrogen levels in the saw- spp. (Chang and Hayes, 1978). et al., 2012). Incubation tem- has been discarded, indicating dust were low, which may be a The substrates formulated perature was 35°C. After 25 that this residue is readily limiting factor for decay. with S. amara sawdust provided days, the colonized substrates available. The the highest radial mycelial growth (cm/day) of P. strigellus (p<0.01; Figure 2). How- ever, both rice bran and TFS supplementation improved the colony vigor as compared with the control S. amara sawdust. Al- though the mycelial growth on PPIS formulations supplemented Figure 2. Effect of supplementing Bactris gasipaes internal sheath with rice bran Figure 1. Effect of substrate humidity on Panus strigel- and Simarouba amara sawdust on Panus strigellus mycelial growth and TFS was lus mycelial growth. Substrate formulation was a mix- at 35°C. C: control, R: rice bran, TFS: Astrocaryum aculeatum lower, the colony ture of Hymenolobium petraeum sawdust and rice bran fruit shell, PPIS: B. gasipaes internal sheath. Substrate composition vigor was at the (10:1 w/w). Data shown represent the average of five was 10:1 and 5:1 (sawdust:supplement, w/w). Data shown represent highest level; replicates and two repetitions of mycelial growth (cm/ the average mycelial growth (cm/day) in five replications and two day). Means indicated by the same letter are not sig- repetitions. Means indicated by the same letter(s) are not signifi- thus, in the sec- nificantly different (p<0.01) by the Tukey test. Colony cantly different (p<0.01) by the Tukey test. Colony vigor levels are ond experiment vigor levels are +: thin, ++: medium, and +++: dense. +: thin, ++: medium, and +++: dense. PPIS supplemen-

OCT 2013, VOL. 38 Nº 10 735 the spawn production of P a n u s mento e produção em experimen- strigellus. tos com pupunheira para palmito. Acta Amaz. 30: 349-362. Acknowledgments Donini LP, Bernardi E, Minotto E, Nascimento JS (2005) Desen- volvimento in vitro de Pleurotus The authors thank Kaoru spp. sob a influência de diferentes Yuyama for providing specimens substratos de dextrose. Arq. Inst. of B. gasipaes. This research Biol. 72: 331-338. was financed by Fundação de Eira FC, Meirelles WF, Paccola-Meire- Amparo à Pesquisa do Estado lles LD (2005) Shiitake production do Amazonas (FAPEAM), the in corncob substrates. Rev. Bras. Coordenação de Aperfeiçoa- Milho Sorgo 4: 141-148. mento de Pessoal de Nível Supe- Fan L, Soccol AT, Pandey A, Van- rior (CAPES) by the PNADB denberghe LPS, Soccol CR (2006) Effect of caffeine and Program and the Centro de Es- tannins on cultivation and fruc- tudos Integrados de Biodivers- Figure 3. Effect of supplementing sawdust with Bactris gasipaes internal tification of Pleurotus on coffee sheath on Panus strigellus mycelial growth at 35°C. R: rice bran, PPIS: idade Amazônica (CENBAM). husks. Braz. J. Microbiol. 37: B. gasipaes internal sheath. Substrate was composed of 10:1 and 1:1 RV-I acknoledges a scholarship 420-424. (sawdust:supplement, w/w). Data shown represent the average mycelial from Conselho Nacional de De- IDAM (2012) Cultura: Pupunha (pal- growth (cm/day) in five replicates and two repetitions. Means indicated senvolvimento Científico e Tec- mito). Relatório de acompanha- by the same letter are not significantly different (p<0.01) by the Tukey nológico (CNPq), Brazil. mento trimestral: Janeiro-Dezem- test. Colony vigor levels are +: thin, ++: medium, and +++: dense. bro 2012. Instituto de Desenvolvi- mento Agropecuário e Florestal REFERENCES Sustentável do Estado do Amazo- nas, Brasil. 122 pp. tation was used. 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