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A Duplex PCR Assay for Rapid Detection of Phytophthora nicotianae and Thielaviopsis basicola

文献类型： 外文期刊

作者： Liu, Na 1 ; Jiang, Shijun 2 ; Feng, Songli 1 ; Shang, Wenyan 1 ; Xing, Guozhen 1 ; Qiu, Rui 3 ; Li, Chengjun 3 ; Li, Shuj 1 ;

作者机构： 1.Henan Agr Univ, Coll Life Sci, Zhengzhou 450002, Henan, Peoples R China

2.Henan Agr Univ, Coll Plant Protect, Zhengzhou 450002, Henan, Peoples R China

3.Henan Acad Agr Sci, Key Lab Green Preservat & Control Tobacco Dis & P, Tobacco Res Inst, Xuchang 461000, Peoples R China

关键词： duplex PCR; molecular detection; Phytophthora nicotianae; Thielaviopsis basicola; tobacco

期刊名称：PLANT PATHOLOGY JOURNAL （ 影响因子：1.795； 五年影响因子：2.314 ）

ISSN： 1598-2254

年卷期： 2019 年 35 卷 2 期

页码：

收录情况： SCI

摘要： A duplex PCR method was developed for simultaneous detection and identification of tobacco root rot pathogens Phytophthora nicotianae and Thielaviopsis basicola. The specific primers for P. nicotianae were developed based on its internal transcribed spacer (ITS) regions of ribosomal gene, ras gene and hgd gene, while the specific primers for T. basicola were designed based on its ITS regions and beta-tubulin gene. The specificity of the primers was determined using isolates of P. nicotianae, T. basicola and control samples. The results showed that the target pathogens could be detected from diseased tobacco plants by a combination of the specific primers. The sensitivity limitation was 100 fg/mu l of pure genomic DNA of the pathogens. This new assay can be applied to screen out target pathogens rapidly and reliably in one PCR and will be an important tool for the identification and precise early prediction of these two destructive diseases of tobacco.

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