从混合线虫样品和植物组织中直接检测香蕉穿孔线虫的ITS-PCR方法

doi:10.3864/j.issn.0578-1752.2011.19.008

Abstract

Abstract: 【Objective】 The objective of this study is to build an ITS-PCR (internal transcribed spacer-polymerase chain reaction) method for detecting Radopholus similis (Cobb) Thorne from mixtures of different nematodes, or tissues of Musa spp. and Anthurium andraeanum. 【Method】 A specific primer pair was designed according to ITS region of rDNA from R. similis using Primer Premier 5.0. A single-step PCR method was used to specially detect the target ITS-rDNA of R. similis. 【Result】 The specificity of the designed primers was confirmed that a 518 bp PCR product was amplified from 7 populations of R. similis, not from the other 16 species including 17 populations. Using the established DNA extraction method and PCR system by the specific primers, the target ITS-rDNA fragment was specially amplified from DNA templates of the mixture of individuals of R. similis with species of Pratylenchus, Helicotylenchus, Rotylenchulus, Meloidogyne, Ditylenchus, Tylenchorhynchus, Hoplolaimus, Filenchus, Aphelenchoides, and Rhabditida, and also from the DNA mixtures of at least 3 individuals of R. similis with root tissues (2 cm long, about 0.1 g) of banana and Anthurium andraeanum. 【Conclusion】 The ITS-PCR method established in this study was proven to be a reliable and useful technique for fast and accurate detecting of R. similis from mixed nematode samples and mixtures of root tissues of banana or A.andraeanum.

Key words:

Radopholus similis, rDNA-ITS region specific primers, nematodes mixture, plant tissues, PCR detection

LIU Yi-Fan, XU Chun-Ling, ZHANG Chao, SU Xiu-Min, XIE Hui. Direct PCR Detection of Radopholus similis from Plant Tissues and Mixed Sample of Nematodes by ITS Specific Primers[J].Scientia Agricultura Sinica, 2011, 44(19): 3991-3998.

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Direct PCR Detection of Radopholus similis from Plant Tissues and Mixed Sample of Nematodes by ITS Specific Primers

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