数字农科院2.0

Species-specific PCR-based assays for identification and detection of Botryosphaeriaceae species causing stem blight on blueberry in China

文献类型： 外文期刊

作者： Xu Cheng-nan;Zhang Hong-jun;Chi Fu-mei;Ji Zhi-rui;Dong Qing-long;Zhou Zong-shan;Xu Cheng-nan;Cao Ke-qiang

作者机构：

关键词： blueberry;stem blight;PCR;Botryosphaeriaceae;species-specific primer

期刊名称： JOURNAL OF INTEGRATIVE AGRICULTURE

ISSN： 2095-3119

年卷期： 2016 年 15 卷 3 期

页码：

收录情况： SCI

摘要： Botryosphaeriaceae species are important causal agents of blueberry stem blight worldwide. Blueberry stem blight has become an important disease, potentially affecting the quality and production of blueberries in China. It is difficult and time-consuming to identify at the species level using morphological methods. The aim of this study was to develop polymerase chain reaction (PCR) assays for the diagnosis and early detection of latent infections of blueberry stems by Botryosphaeria spp. Species-specific primers, based on the ribosomal DNA internal transcribed spacer region and beta-tubulin gene, were designed and selected for use in PCR assays. Three primer pairs, Lt347-F/R for Lasiodiplodia theobromae, Np304-FIR for Neofusicoccum parvum and FaF/Bt2b for Botryosphaeria dothidea, successfully amplified specific PCR fragments of different sizes on pure cultures or from blueberry stems inoculated and naturally infected blueberry plants with three pathogens, respectively. These primers did not amplify any PCR fragments from other blueberry stem disease-associated pathogens, such as Phomopsis spp. and Pestalotiopsis spp. This PCR protocol could detect as low as 100 pg to 1 ng of purified fungal DNA. This PCR-based protocol could be used for the diagnosis and detection of these pathogens from pure cultures or from infected blueberry plants.

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