Development of a nested-PCR assay for the rapid detection of Pilidiella granati in pomegranate fruit

文献类型： 外文期刊

第一作者： Yang, Xue

作者： Yang, Xue;Hameed, Uzma;Zhang, Ai-Fang;Zang, Hao-Yu;Gu, Chun-Yan;Chen, Yu;Yang, Xue;Zhang, Ai-Fang;Zang, Hao-Yu;Gu, Chun-Yan;Chen, Yu;Yang, Xue;Zhang, Ai-Fang;Zang, Hao-Yu;Gu, Chun-Yan;Chen, Yu;Hameed, Uzma;Xu, Yi-Liu;Xu, Yi-Liu

作者机构：

期刊名称：SCIENTIFIC REPORTS （ 影响因子：4.379； 五年影响因子：5.133 ）

ISSN： 2045-2322

年卷期： 2017 年 7 卷

页码：

收录情况： SCI

摘要： Pilidiella granati, a causal agent of twig blight and crown rot of pomegranate, is an emerging threat that may cause severe risk to the pomegranate industry in the future. Development of a rapid assay for the timely and accurate detection of P. granati will be helpful in the active surveillance and management of the disease caused by this pathogen. In this study, a nested PCR method was established for the detection of P. granati. Comparative analysis of genetic diversity within 5.8S rDNA internal transcribed spacer (ITS) sequences of P. granati and 21 other selected fungal species was performed to design species-specific primers (S1 and S2). This primer pair successfully amplified a 450 bp product exclusively from the genomic DNA of P. granati. The developed method can detect 10 pg genomic DNA of the pathogen in about 6 h. This technique was successfully applied to detect the natural infection of P. granati in the pomegranate fruit. The designed protocol is rapid and precise with a high degree of sensitivity.

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