title: Design and evaluation of PCR primers for denaturing gradient gel electrophoresis analysis of plant parasitic and fungivorous nematode communities.
abstract: A PCR-DGGE primer pair, Tyl2F-Tyl4R, specific to plant parasitic and fungivorous nematodes was designed based on the 18S rRNA gene. The results of community analysis using the primers showed that they are specific to the order Tylenchida. This primer pair detected species belonging to Tylenchida with high sensitivity and high resolution. The number of detected species of plant parasitic and fungivorous nematodes and their band intensity were much improved compared with PCR-DGGE analysis using the SSU18A-SSU9R primer, which is commonly used for nematode community analysis. It was confirmed that using a group-specific primer was effective for nematode community analysis with PCR-DGGE.
authors.affiliation: Upland Farming Research Division, NARO Hokkaido Agricultural Research Center, Shinsei-minami 9–4, Memuro-cho,Kasai-gun, Hokkaido 082–0071, Japan. kushida@affrc.go.jp
authors.forename: Atsuhiko
authors.initials: A
authors.lastname: Kushida
mesh_terms: D000818:Animals; D044822:Biodiversity; D017931:DNA Primers / Q000235:genetics*; D058645:Denaturing Gradient Gel Electrophoresis / Q000379:methods*; D008969:Molecular Sequence Data; D010944:Plants / Q000469:parasitology*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012337:RNA, Ribosomal, 18S / Q000235:genetics; D017422:Sequence Analysis, DNA; D017187:Tylenchida / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification
chemical_list: D017931:DNA Primers; D012337:RNA, Ribosomal, 18S