Transformation of Phomopsis asparagi with green fluorescent protein using protoplasts

文献类型： 外文期刊

第一作者： Zhang, Yueping

作者： Zhang, Yueping;Qu, Huaxiang;Zhao, Ping;Wu, Luofa;Zhou, Jingsong;Tang, Yongping;Luo, Shaochun;Chen, Guangyu;Qu, Huaxiang

作者机构：

关键词： green fluorescent protein; host-pathogen interaction; Phomopsis asparagi; protoplast transformation; transformants

期刊名称：CANADIAN JOURNAL OF PLANT PATHOLOGY （ 影响因子：2.442； 五年影响因子：1.993 ）

ISSN： 0706-0661

年卷期： 2018 年 40 卷 2 期

页码：

收录情况： SCI

摘要： Phomopsis asparagi is a fungal pathogen that causes destructive stem blight disease in garden asparagus worldwide. The host-pathogen interactions and molecular mechanisms of its pathogenicity are poorly understood. In this study, a protoplast transformation approach mediated by polyethylene glycol (PEG) was established for P. asparagi, and green fluorescent protein (GFP) expression transformants were successfully obtained. The transformants were mitotically stable when grown on potato dextrose agar (PDA) plates amended with 100 mu g mL(-1) of hygromycin B after five successive subcultures without selective pressure. All of the analysed transformants were further confirmed by polymerase chain reaction (PCR) amplification with primers specific to the hygromycin B phosphotransferase (hph) gene and Southern blotting. Moreover, these transformants showed similar pathogenicity on garden asparagus spears compared with the wild type, although a difference in growth rate was observed in oatmeal agar (OA) plates. These GFP expression transformants will serve as useful tools for investigating host penetration and the invasive growth of this devastating fungal pathogen, as well as defining the nature of the host-pathogen interaction.

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