Polymorphism in the internal transcribed spacer (ITS) of the ribosomal DNA of 26 isolates of ectomycorrhizal fungi

Genetics and Molecular Biology

Eliane A. Gomes²  Maria Catarina M. Kasuya¹  Everaldo G. De Barros¹  Arnaldo C. Borges¹  Elza F. Araújo¹ 

[1] ,Embrapa Milho e Sorgo Núcleo de Biologia Aplicada Sete Lagoas MG ,Brazil

关键词: DNA fingerprinting;    ectomycorrhizal fungi;    internal transcribed spacer (ITS);    PCR-RFLP;    ribosomal DNA (rDNA);   

DOI  :  10.1590/S1415-47572002000400018

来源: SciELO

【 摘 要 】

Inter- and intraspecific variation among 26 isolates of ectomycorrhizal fungi belonging to 8 genera and 19 species were evaluated by analysis of the internal transcribed sequence (ITS) of the rDNA region using restriction fragment length polymorphism (RFLP). The ITS region was first amplified by polymerase chain reaction (PCR) with specific primers and then cleaved with different restriction enzymes. Amplification products, which ranged between 560 and 750 base pairs (bp), were obtained for all the isolates analyzed. The degree of polymorphism observed did not allow proper identification of most of the isolates. Cleavage of amplified fragments with the restriction enzymes Alu I, Hae III, Hinf I, and Hpa II revealed extensive polymorphism. All eight genera and most species presented specific restriction patterns. Species not identifiable by a specific pattern belonged to two genera: Rhizopogon (R. nigrescens, R. reaii, R. roseolus, R. rubescens and Rhizopogon sp.), and Laccaria (L. bicolor and L. amethystea). Our data confirm the potential of ITS region PCR-RFLP for the molecular characterization of ectomycorrhizal fungi and their identification and monitoring in artificial inoculation programs.

【 授权许可】

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