Characterisation and identification of arbuscular mycorrhizal fungi species by PCR/RFLP analysis of the rDNA internal transcribed spacer (ITS)

Annals of Microbiology (Impact Factor: 1.55). 01/2008; 58(2):341-344. DOI: 10.1007/BF03175340

ABSTRACT The polymerase chain reaction coupled with restriction fragment length polymorphism analysis (PCR/RFLP) was applied to distinguish
arbuscular mycorrhizal fungi (AMF) species from an impacted semiarid soil. The ITS1-5.8S-ITS2 region was amplified with the
primers ITS1 and ITS4 and the products of amplification were digested with the restriction enzymes HinfI, MboI and AluI. The
obtained banding pattern, except for the AluI, allowed the distinction at the molecular level of the AMF species:Paraglomus occultum, Glomus mosseae, Glomus intraradices andGlomus etunicatum. The results showed that this technique has a potential to be used as a marker to differentiate AMF species with high phylogenetic

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    ABSTRACT: Resumo – O objetivo deste trabalho foi caracterizar fenotípica e genotipicamente isolados de espécies de fungos micorrízicos arbusculares (FMA) mantidos em cultura pura e avaliar a aplicabilidade da técnica PCR-DGGE desenvolvida para Gigaspora, na identificação molecular de espécies de FMA pertencentes a outros gêneros. A caracterização fenotípica das espécies foi realizada de acordo com critérios morfológicos, descritos pela taxonomia, e com uso de descrições originais das espécies presentes na literatura especializada. A análise genotípica foi feita com base na discriminação específica da região V9 do 18S rDNA, que permitiu a diferenciação das espécies e não revelou qualquer diferença entre os isolados geográficos de Glomus clarum, e entre os de Glomus etunicatum. Isto indica a aplicabilidade da técnica para a avaliação da pureza genética e discriminação de espécies de FMA. Termos para indexação: caracterização morfológica, identificação molecular, iniciadores, Glomeromycota, taxonomia.
    01/2010; 458:806-896.
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    ABSTRACT: The objective of this work was to characterize phenotypically and genotypically isolates of arbuscular mycorrhizal fungi (AMF) maintained in pure culture and to evaluate the applicability of PCR-DGGE analysis, developed for Gigaspora, for molecular identification of AMF species belonging to other genres. The species phenotypic characterization was done according to morphological criteria, as described by taxonomy, and according to original descriptions of species published in the specialized literature. The genotypic analysis was made through specific discrimination of the V9 region in the 18S rDNA, which allowed the distinction of species and showed no difference among geographical isolates of Glomus clarum, and among those of Glomus etunicatum. This indicates the applicability of this technique for assessment of genetic purity and discrimination of AMF species.
    Pesquisa Agropecuária Brasileira 08/2010; 45(8):886-896. · 0.66 Impact Factor

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