Abstract:This paper describes for the first time in vitro mycorrhization between the two wild edible boletes (Boletus edulis and Suillus sibiricus) with Pinus gerardiana. The synthesis was carried out in a controlled growth chamber using peat, vermiculite, fungal medium and mycelial inoculum of each fungi in test tubes. The test tubes were regularly observed for mycorrhization. The seedlings of P. gerardiana were picked after five months of inoculation to examine symbiotic association between its root system with B. ed… Show more
The present investigations are aimed to synthesize in vitro ectomycorrhizae between Pinus gerardiana and two gilled ectomycorrhizal (ECM) mushrooms (Amanita ceciliae and Lactarius sanguifluus). To carry out in vitro synthesis, pure cultures of ECM mushrooms (A. ceciliae and L. sanguifluus) were isolated on Potato Dextrose Agar (PDA) and Modified Melin-Norkans (MMN) Medium respectively. The synthesis was achieved successfully in the surfaced sterilized seedlings of P. gerardiana geminated under aseptic conditions by using vermiculite, peat, medium for ECM fungi and inoculum of each fungus in the test tubes. Mycorrhization was checked periodically in the test tubes. P. gerardiana seedlings were lifted from test tubes after five months to observe ectomycorrhizae formation on the root system with A. ceciliae and L. sanguifluus. The synthesized ectomycorrhizae were dark brown in case of A. ceciliae whereas in case of L. sanguifluus the colour of ECM roots was yellowish brown. Anatomy of synthesized ectomycorrhizae with both ECM fungi showed fully developed fungal mantle and Hartig net. The seedlings with ECM synthesis showed a significant effect on the growth and development.
The present investigations are aimed to synthesize in vitro ectomycorrhizae between Pinus gerardiana and two gilled ectomycorrhizal (ECM) mushrooms (Amanita ceciliae and Lactarius sanguifluus). To carry out in vitro synthesis, pure cultures of ECM mushrooms (A. ceciliae and L. sanguifluus) were isolated on Potato Dextrose Agar (PDA) and Modified Melin-Norkans (MMN) Medium respectively. The synthesis was achieved successfully in the surfaced sterilized seedlings of P. gerardiana geminated under aseptic conditions by using vermiculite, peat, medium for ECM fungi and inoculum of each fungus in the test tubes. Mycorrhization was checked periodically in the test tubes. P. gerardiana seedlings were lifted from test tubes after five months to observe ectomycorrhizae formation on the root system with A. ceciliae and L. sanguifluus. The synthesized ectomycorrhizae were dark brown in case of A. ceciliae whereas in case of L. sanguifluus the colour of ECM roots was yellowish brown. Anatomy of synthesized ectomycorrhizae with both ECM fungi showed fully developed fungal mantle and Hartig net. The seedlings with ECM synthesis showed a significant effect on the growth and development.
En México, los hongos ectomicorrizógenos silvestres (HEcM) son un importante recurso forestal no maderable y son aprovechados tradicionalmente para su autoconsumo y venta. La conservación in vitro de cepas de HEcM es más compleja que en especies saprófitas, debido a las relaciones fisiológicas existentes con sus hospederos. En este trabajo se presenta un método de crioconservación de cepas de HEcM utilizando vectores no convencionales y la posterior evaluación de su viabilidad in vitro. Para ello, cinco cepas de HEcM (Boletus aff. edulis, Boletus sp., Helvella sp. y Lactarius indigo [2]) fueron criogenizadas (-196 °C) durante 30 días, utilizando como vectores acículas de pino y semillas de mijo estériles. Posteriormente, se determinó la capacidad de dos de las cepas recuperadas para formar morfotipos de ectomicorrizas in vitro inoculados en plántulas de Pinus montezumae. Los resultados de la primera etapa mostraron altos porcentajes de recuperación de los micelios criogenizados en ambos vectores (98% - 100%). En cuanto a la micorrización in vitro, las plántulas no micorrizadas (control) no sobrevivieron, mientras que las micorrizadas mostraron valores de sobrevivencia entre 33% y 100%, con incremento de la longitud de sus tallos. Además, se observó la formación de morfotipos dicotómicos de color ocre oscuro, así como el desarrollo del manto fúngico en las raíces micorrizadas después de cuatro meses de inoculación. El método empleado para la criogenización de las cepas de HEcM posibilita la conservación de los micelios por períodos prolongados, manteniendo activa su capacidad de micorrización.
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