Semisolid culture medium improves mycelial recovery of Agaricus subrufescens cryopreserved in cereal grains
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This study aimed to evaluate the effects of the solid and semisolid culture medium on the mycelial viability of A. subrufescens after 5-year cryopreservation at − 70 °C. Mycelia were grown in three types of whole or ground grains, with or without 5% glycerol addition in the substrate and/or in a cryotube. After 5 years of cryopreservation at − 70 °C, every treatment was thawed and recovered in malt extract culture medium with 15 (solid culture medium) or 5 g L−1 (semisolid culture medium) of agar. The semisolid recovery culture medium increased the mycelial viability recovery capacity of A. subrufescens cryopreserved for 5 years in grains with glycerol only in the cryotube, and specifically with medium-hard wheat grain without glycerol addition at all. Agar-based substrates such as malt extract agar, agar-ground grain, or the one with glycerol addition to the substrate were not effective to keep the mycelial viability, regardless of the recovery culture medium consistency. Hard and medium-hard endosperm wheat grains or hard endosperm rye grains with addition of glycerol as cryoprotectant only to the cryotube were effective to cryopreserve the fungus for 5 years without cryoprotectant addition in the substrate.
KeywordsAgaricus blazei Semisolid culture medium Water activity Mycelial cryopreservation Thawing
The authors thank Universidade Paranaense, Programa de Pós-graduação em Biotecnologia Aplicada à Agricultura, Coordenação de Aperfeiçoamento de Pessoal de Nível Superior—Brasil (CAPES)—finance code 001, Fundação Araucária, and Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) for the financial support and fellowship.
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