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Molecular approach to characterize ectomycorrhizae fungi from Mediterranean pine stands in Portugal

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Stone pine (Pinus pinea L.), like other conifers, forms ectomycorrhizas (ECM), which have benefi cial impact on plant growth in natural environments and forest ecosystems. An in vitro co-culture of stone pine microshoots with pure mycelia of isolated ECM sporocarps was used to overcome the root growth cessation not only in vitro but also to improve root development during acclimation phase. Pisolithus arhizus (Scop.) Rauschert and Lactarius deliciosus (L. ex Fr.) S.F. Gray fungi, were col lected, pure cultured and used in in vitro co-culture with stone pine microshoots. Samples of P. arhizus and L. deliciosus for the in vitro co-cultures were collected from the pine stands southwest Portugal. The in situ characterization was based on their morphotypes. To confirm the identity of the collected material, ITS amplification was applied using the pure cultures derived from the sporo carps. Additionally, a molecular profile using PCR based genomic fingerprinting comparison was executed with other genera of Basidiomycetes and Ascomycetes. Our results showed the effective ness of the techniques used to amplify DNA polymorphic sequences, which enhances the characte rization of the genetic profile of ECM fungi and also provides an option to verify the fungus identity at any stage of plant mycorrhization.

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Pisolithus arhizus Lactarius deliciosus Pinus pinea M13-PCR ITS . Escola Superior de Tecnologias e Gestão

Citation

Ragonezi, C., Caldeira, A. T., Rosário Martins, M., Salvador, C., Santos-Silva, C., Ganhão, E., ... & Zavattieri, A. (2013). Molecular approach to characterize ectomycorrhizae fungi from Mediterranean pine stands in Portugal. Brazilian Journal of Microbiology, 44(2), 663-670.

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Sociedade Brasileira de Microbiologia