An efficient Agrobacterium-mediated transformation method for the edible mushroom Hypsizygus marmoreus

文献类型： 外文期刊

第一作者： Zhang, Jin Jing

作者： Zhang, Jin Jing;Shi, Liang;Sun, Yun Qi;Zhao, Ming Wen;Ren, Ang;Feng, Zhi Yong;Zhang, Jin Jing;Chen, Hui;Sun, Yun Qi;Chen, Ming Jie;Wang, Hong;Feng, Zhi Yong;Zhang, Jin Jing;Chen, Hui;Sun, Yun Qi;Chen, Ming Jie;Wang, Hong;Feng, Zhi Yong

作者机构：

关键词： Agrobacterium-mediated transformation;Hypsizygus marmoreus;gpd;Promoter

期刊名称：MICROBIOLOGICAL RESEARCH （ 影响因子：5.415； 五年影响因子：6.038 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： Hypsizygus marmoreus is one of the major edible mushrooms in East Asia. As no efficient transformation method, the molecular and genetics studies were hindered. The glyceraldehyde-3-phosphate dehydrogenase (GPD) gene of H. marmoreus was isolated and its promoter was used to drive the hygromycin B phosphotransferase (HPH) and enhanced green fluorescent protein (EGFP) in H. marmoreus. Agrobacterium tumefaciens-mediated transformation (ATMT) was successfully applied in H. marmoreus. The transformation parameters were optimized, and it was found that co-cultivation of bacteria with protoplast at a ratio of 1000:1 at a temperature of 26 degrees C in medium containing 0.3 mM acetosyringone resulted in the highest transformation efficiency for Agrobacterium strain. Besides, three plasmids, each carrying a different promoter (from H. marmoreus, Ganoderma lucidum and Lentinula edodes) driving the expression of an antibiotic resistance marker, were also tested. The construct carrying the H. marmoreus gpd promoter produced more transformants than other constructs. Our analysis showed that over 85% of the transformants tested remained mitotically stable even after five successive rounds of subculturing. Putative transformants were analyzed for the presence of hph gene by PCR and Southern blot. Meanwhile, the expression of EGFP in H. marmoreus transformants was detected by fluorescence imaging. This ATMT system increases the transformation efficiency of H. marmoreus and may represent a useful tool for molecular genetic studies in this mushroom species

分类号： Q

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