Development of a specific PCR assay for the detection of Rhizoctonia solani AG 1-IB using SCAR primers

被引:44
作者
Grosch, R.
Schneider, J. H. M.
Peth, A.
Waschke, A.
Franken, P.
Kofoet, A.
Jabaji-Hare, S. H.
机构
[1] Grossbeeren Erfurt eV, IGZ, Inst Vegetable & Ornamental Crops, Erfurt, Germany
[2] McGill Univ, Dept Plant Sci, Montreal, PQ H3A 2T5, Canada
关键词
disease diagnosis; phylogenetic analysis; Rhizoctonia solani; subgroup identification;
D O I
10.1111/j.1365-2672.2006.03125.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Aims: The aim of this study was to develop a specific and sensitive identification method for Rhizoctonia solani AG 1-IB isolates based on phylogenetic relationships of R. solani AG-1 subgroups using rDNA-internal transcribed spacer (rDNA-ITS) sequence analysis. Methods and Results: A neighbour-joining tree analysis of 40 rDNA-ITS sequences demonstrated that R. solani AG-1 isolates cluster separately in six subgroups IA, IB, IC, ID, IE and IF. A molecular marker was generated from a random amplified polymorphic DNA fragment (RAPD). After conversion into a sequence-characterized amplified region (SCAR), a specific primer set for identification of subgroup AG 1-IB was designed for use in a polymerase chain reaction (PCR). The primer pair amplified a single DNA product of 324 bp. Conclusions: R. solani AG-1 subgroups were discriminated by sequence analysis of the ITS region. The designed SCAR primer pair allowed an unequivocal and rapid detection of R. solani AG 1-IB in plant and soil samples. Significance and Impact of the Study: Sequence analysis of the rDNA-ITS region can be used for differentiation of subgroups within AG-1. The use of the developed SCAR primer set allowed a reliable and fast identification of R. solani AG 1-IB and provides a powerful tool for disease diagnosis.
引用
收藏
页码:806 / 819
页数:14
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