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

Abstract

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.

Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi

“…Intragenomic variation, such as the existence of multiple paralogous or nonorthologous copies within single fruiting bodies of basidiomycetes (54,55) and ascomycetes (56) or within axenic cultures (51), may lead to higher estimates of infraspecific variability (57,58) or generation of barcodes that act only as representative sequences of multiple variable repeats (59,60). Highly variable lengths and high evolutionary rates for the nuclear ribosomal cistron in species of Cantharellus, Tulasnella (Cantherellales, Basidiomycota) (61)(62)(63), and some lichens (53) may provide challenges for sequencing and analysis.…”

Assessing the Genetic Diversity of <i>Alternaria Bataticola</i> in South Africa using Molecular Markers

“…The rDNA ITS2 region was amplified following a modified protocol described by Gomes et al [24] using the two primers (5'-TCCGTAGGTGAACCTGCGG-3'; '5'-GCTGCGTTCTTCATCGATGC-3') [25]. The PCR products were sequenced at Inqaba Biotech (Pretoria, South Africa).…”

Sekuen Internal Transcribed Spacer (ITS) DNA ribosomal Oncobasidium theobromae dan jamur sekerabat pembanding Internal Transcribed Spacer (ITS) sequences of ribosomal DNA Oncobasidium theobromae and other related fungi as comparison

“…Organisme eukaryotik mempunyai dua daerah ITS; ITS-1 terletak di antara 18S gen dan 5.8S gen, dan ITS-2 terletak di antara 5.8S dan 28S gen. Ketiga gen ribosom tersebut mempunyai tingkat konservasi yang sangat tinggi (Gomes et al, 2002;O'Brien et al, 2005). Daerah ITS biasanya mengalami perubahan atau mutasi sehingga dapat berbeda atau bervariasi di antara spesies.…”