title: Molecular approach to characterize ectomycorrhizae fungi from Mediterranean pine stands in Portugal.
abstract: Stone pine (Pinus pinea L.), like other conifers, forms ectomycorrhizas (ECM), which have beneficial 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 collected, 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 sporocarps. Additionally, a molecular profile using PCR based genomic fingerprinting comparison was executed with other genera of Basidiomycetes and Ascomycetes. Our results showed the effectiveness of the techniques used to amplify DNA polymorphic sequences, which enhances the characterization of the genetic profile of ECM fungi and also provides an option to verify the fungus identity at any stage of plant mycorrhization.
authors.affiliation: Instituto de Ciências Agrárias e Ambientais Mediterrânicas, Universidade de Évora, Núcleo da Mitra, Évora, Portugal.
authors.forename: Carla
authors.initials: C
authors.lastname: Ragonezi
authors.forename: A Teresa
authors.initials: AT
authors.lastname: Caldeira
authors.forename: M Rosário
authors.initials: MR
authors.lastname: Martins
authors.forename: Cátia
authors.initials: C
authors.lastname: Salvador
authors.forename: Celeste
authors.initials: C
authors.lastname: Santos-Silva
authors.forename: Elsa
authors.initials: E
authors.lastname: Ganhão
authors.forename: Krystyna
authors.initials: K
authors.lastname: Klimaszewska
authors.forename: Amely
authors.initials: A
authors.lastname: Zavattieri
mesh_terms: D016000:Cluster Analysis; D004271:DNA, Fungal / Q000737:chemistry / Q000235:genetics; D018744:DNA, Plant / Q000737:chemistry / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D038821:Mycorrhizae / Q000145:classification* / Q000235:genetics / Q000254:growth & development / Q000302:isolation & purification*; D010802:Phylogeny; D028223:Pinus / Q000382:microbiology*; D011174:Portugal; D017422:Sequence Analysis, DNA
chemical_list: D004271:DNA, Fungal; D018744:DNA, Plant; D021903:DNA, Ribosomal Spacer
keywords: ITS; Lactarius deliciosus; M13-PCR; Pinus pinea; Pisolithus arhizus