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    Plant pathology发表大豆根腐病检测靶标

      

    题目:A Phytophthora conserved transposon-like DNA element is a potential target for soybean root rot disease diagnosis

    作者:Ting-ting Dai, Jun Meng, ChenChen Lu, SuoMeng Dong, WenWu Ye, XiaoBo Zheng ,YuanChao Wang*, Department of Plant Pathology, Nanjing Agricultural University, Nanjing 210095,China

    研究摘要: Phytophthora sojae is a devastating pathogen that causes soybean Phytophthora root rot. In this study, a transposon-like fragment with multiple copies in the P. sojae genome was identified as a suitable detection target for P. sojae. The PCR primers TrapF1/TrapR1 were designed based on unique sequences derived from this transposon-like sequence. An approximately 276-bp DNA fragment was amplified using the specific primer pair TrapF1 and TrapR1. The specificity of this primer pair was evaluated against 120 isolates of P. sojae, 58 isolates from 25 other Phytophthora spp., 54 isolates of Pythium spp., and isolates of true fungi. Tests with P. sojae genomic DNA determined that a detection sensitivity of 10pg and 10fg DNA in standard PCR (TrapF1/TrapR1) and nested PCR(TrapF1/TrapR1 and TrapF2/TrapR2), respectively, was achieved. Meanwhile, PCR with TrapF1/TrapR1 primers detected the pathogen at the level of a single oospore, and even one zoospore. These primers also have been proved to be efficient in detecting pathogens from diseased soybean tissue and residues. In addition, Real-time quantitative PCR (qPCR) assays coupled with TrapF1/TrapR1 primers were developed to detect and quantify the pathogen. The results demonstrated that the TrapF1/TrapR1 and TrapF2/TrapR2 primers-based PCR assay provides a rapid and sensitive tool for the detection of P. sojae in plants and in production fields.

     


    发布时间: 2012-05-29 11:12 浏览次数:   【 打 印 】【 关 闭
     
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