基于Ypt1基因为靶标的木香疫病病组织及病田土壤分子检测技术(英文)

Molecular Detection of Phytophthora tentaculatain Infected Aucklandia lappa Tissues andSoil on the Basis of Ypt1 Gene

A species-specific PCR assay was established for rapid and accurate detection of the oomycete pathogen Phytophthora tentaculata in diseased plant tissues and infected soil.A pair of species-specific primers Pt1/Pt2 were designed on the basis of Ras-related protein(Ypt1) gene sequences of the Phytophthora species.PCR amplification with the Pt primers resulted in a 386 bp product only from isolates of P.tentaculata.The detection threshold with Pt primers was 100 pg of genomic DNA.A nested PCR procedure was developed using Ypt1F/Ypt1R as the first-round amplification primers and Pt1/Pt2 as the second-round primers,which increased the detection sensitivity 100-fold to 1 pg.PCR using these Pt primers can also be used to detect P.tentaculata in naturally infected plant tissues and soil.The PCR-based method developed in this study provides a rapid and sensitive tool for detection of P.tentaculata.

A species-specific PCR assay was established for rapid and accurate detection of the oomycete pathogen Phytophthora tentaculata in diseased plant tissues and infected soil. A pair of species-specific primers Ptl/Pt2 were designed on the basis of Ras-related protein （ Yptl ） gene sequences of the Phytophthora species. PCR amplification with the Pt primers resulted in a 386 bp product only from isolates of P. tentaculata. The detection threshold with Pt primers was 100 pg of genomic DNA. A nested PCR procedure was developed using YptlF/YptlR as the first-round amplification primers and Pt1/Pt2 as the second-round primers, which increased the detection sensitivity 100-fold to 1 pg. PCR using these Pt primers can also be used to detect P. tentaculata in naturally infected plant tissues and soil. The PCR-based method developed in this study provides a rapid and sensitive tool for detection of P. tentaculata.