Open AccessPCR amplification and characterization of the intergenic spacer region of the ribosomal DNA in Pyrenophora graminea
Author(s) -
Pecchia Susanna,
Mercatelli Elisabetta,
Vannacci Giovanni
Publication year - 1998
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1998.tb13178.x
Subject(s) - ribosomal dna , intergenic region , ribosomal intergenic spacer analysis , biology , ribosomal rna , dna , genetics , gene , phylogenetics , genome
Successful amplification of the whole intergenic spacer region of the nuclear ribosomal repeat (IGS) in Pyrenophora graminea was obtained with a PCR‐based assay. Single amplification products showed length differences. Depending on the length of the IGS‐PCR product, ca. 3.8 or 4.4 kb, two groups of isolates could be identified. The RFLP patterns of isolates obtained with the 6‐base cutting enzymes Apa I, Bgl II, Dra I, Eco RV, Hin dIII and Sac I were similar within each group and different between the two groups. Restriction patterns of IGS‐PCR products digested with the 4‐base cutting enzyme Alu I were polymorphic among isolates in spite of their IGS‐PCR product length. In order to characterize the long and short IGS‐PCR products the restriction map is shown. The long product shows an additional Hin dIII site and a Bgl II site that is lacking in the short product. However, the latter shows a Sac I site that is not present in the long IGS‐PCR product. Therefore, the described PCR‐RFLP analysis of the IGS appears to be a useful tool to resolve genetic variation between P. graminea isolates.