Uproszczona metoda molekularnej identyfikacji nicieni z rodzaju Bursaphelenchus, należących do grupy xylophilus, z wykorzystaniem reakcji PCR ze starterami specyficznymi

• Autorzy: Filipiak A. , Tomalak M.

Warianty tytułu

EN

Simplified method for molecular identification of nematodes from genus Bursaphelenchus belonging to the xylophilus group by using the PCR reaction with specific primers

• Języki publikacji: PL

Abstrakty

EN

Both native European species, Bursaphelenchus mucronatus and B. fraudulentus, are relatively common and harmless to trees. They belong to the xylophilus group, which includes also the quarantine pest B. xylophilus – the causal agent of the pine wilt disease. They can be difficult to distinguish morphologically from each other and from B. xylophilus. Therefore, reliable methods of taxonomic identification of these species are therefore of particular interest to plant quarantine services. PCR amplification with specific primers enables rapid and precise species identification, even from a single nematode. In 2004, Matsunaga and Togashi designed specific primers for B. xylophilus and B. mucronatus. The aim of this study was to design specific primers for B. fraudulentus. The specificity of the newly designed primers was tested on eight isolates of B. fraudulentus which originated from different parts in Europe (Austria, Germany, Poland, Russia and Ungarn). For all isolates, PCR amplification resulted in products of identical length of 617 bp, but no PCR products were generated for B. xylophilus and B. mucronatus. The PCR amplification with primer sets specific for B. xylophilus (XF and XR), B. mucronatus (MF and MR) and those for B. fraudulentus (FF and FR) designed in this study resulted in amplicons of different lengths (557, 210 and 617 bp, respectively), which can be easily distinguished in agarose gels.

Słowa kluczowe

PL

Bursaphelenchus; Bursaphelenchus fraudulentus; Bursaphelenchus mucronatus; Bursaphelenchus xylophilus; identyfikacja patogenow; lancuchowa reakcja polimerazy; metody badan; metody molekularne; nicienie chorobotworcze; pasozyty roslin; sekwencje nukleotydowe; startery specyficzne

EN

Bursaphelenchus; Bursaphelenchus fraudulentus; Bursaphelenchus mucronatus; Bursaphelenchus xylophilus; pathogen identification; polymerase chain reaction; research method; molecular method; pathogenic nematode; plant parasite; nucleotide sequence; specific primer

• Wydawca: -
• Czasopismo: Progress in Plant Protection
• Rocznik: 2010
• Tom: 50
• Numer: 1
• Opis fizyczny: s.107-112,fot.,bibliogr.

Twórcy

autor

Filipiak A.

• Instytut Ochrony Roślin – Państwowy Instytut Badawczy, ul.Władysława Węgorka 20, 60-318 Poznań

autor

Tomalak M.

Bibliografia

• Burgermeister W., Metge K., Braasch H., Buchbach E. 2005. ITS-RLFP patterns for differentiation of 26 Bursaphelenchus species (Nematoda: Parasitaphelenchidae) and observations on their distribution. Russian J. Nematol. 13: 29–42.
• Ferris V.R., Ferris J.M., Faghihi J. 1993. Variation in spacer ribosomal DNA in some cystforming species of plant parasitic nematodes. Fundamental and Applied Nematology 16: 177–184.
• Filipiak A., Jakubowska A., Tomalak M. 2010. Species-specific polymerase chain reaction primers for a simple detection of Bursaphelenchus fraudulentus (Nematoda: Parasitaphelenchidae). Nematology 12 (1): 157–160.
• Kanzaki N. 2008. Taxonomy and systematics of the nematode genus Bursaphelenchus (Nematoda: Parasitaphelenchidae). p. 44–66. In: ,,Pine Wilt Disease” (B.G. Zhao, K. Futai, J.R. Sutherland, Y. Takeuchi, eds.). Springer Japan, Tokyo, Japan, 459 pp.
• Matsunaga K., Togashi K. 2004. A simple method for discriminating Bursaphelenchus xylophilus and B. mucronatus by species-specific polymerase chain reaction primer pairs. Nematology 6 (2): 273–277.
• Tomalak M. 2004. Ocena przydatności dostępnych metod do identyfikacji taksonomicznej kwarantannowego nicienia – węgorka sosnowca [Bursaphelenchus xylophilus (Steiner et Bührer) Nickle]. Prog. Plant Protection/Post. Ochr. Roślin 44 (1): 452–461.
• Vrain T.C. 1993. Restriction fragment length polymorphism separates species of the Xiphinema americanum group. J. Nematol. 25: 361–364.