title: PCR amplification and characterization of the intergenic spacer region of the ribosomal DNA in Pyrenophora graminea.
abstract: 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 Apal, BglII, DraI, EcoRV, HindIII and SacI were similar within each group and different between the two groups. Restriction patterns of IGS-PCR products digested with the 4-base cutting enzyme AluI 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 HindIII site and a BglII site that is lacking in the short product. However, the latter shows a SacI 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.
authors.affiliation: Dipartimento di Coltivazione e Difesa delle Specie Legnose, Università degli Studi di Pisa, Italy. specchia@agr.unipi.it
authors.forename: S
authors.initials: S
authors.lastname: Pecchia
authors.forename: E
authors.initials: E
authors.lastname: Mercatelli
authors.forename: G
authors.initials: G
authors.lastname: Vannacci
mesh_terms: D001203:Ascomycota / Q000235:genetics* / Q000302:isolation & purification / Q000472:pathogenicity; D001483:Base Sequence; D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics*; D004275:DNA, Ribosomal / Q000235:genetics*; D014644:Genetic Variation; D001467:Hordeum / Q000382:microbiology; D010935:Plant Diseases / Q000382:microbiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D015183:Restriction Mapping; D013696:Temperature
chemical_list: D017931:DNA Primers; D004271:DNA, Fungal; D004275:DNA, Ribosomal