Establishment of an efficient transformation system for Pleurotus ostreatus

文献类型： 外文期刊

第一作者： Lei, Min

作者： Lei, Min;Wu, Xiangli;Zhang, Jinxia;Huang, Chenyang;Lei, Min;Wang, Hexiang

作者机构：

关键词： Agrobacterium tumefaciens-mediated transformation;Fluorescence assay;Hygromycin;Pleurotus ostreatus

期刊名称：WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY （ 影响因子：3.312； 五年影响因子：3.58 ）

ISSN： 0959-3993

年卷期： 2017 年 33 卷 12 期

页码：

收录情况： SCI

摘要： Pleurotus ostreatus is widely cultivated worldwide, but the lack of an efficient transformation system regarding its use restricts its genetic research. The present study developed an improved and efficient Agrobacterium tumefaciens-mediated transformation method in P. ostreatus. Four parameters were optimized to obtain the most efficient transformation method. The strain LBA4404 was the most suitable for the transformation of P. ostreatus. A bacteria-to-protoplast ratio of 100: 1, an acetosyringone (AS) concentration of 0.1 mM, and 18 h of co-culture showed the best transformation efficiency. The hygromycin B phosphotransferase gene (HPH) was used as the selective marker, and EGFP was used as the reporter gene in this study. Southern blot analysis combined with EGFP fluorescence assay showed positive results, and mitotic stability assay showed that more than 75% transformants were stable after five generations. These results showed that our transformation method is effective and stable and may facilitate future genetic studies in P. ostreatus.

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