Development of a simple and efficient transformation system for the basidiomycetous medicinal fungus Ganoderma lucidum
文献类型: 外文期刊
第一作者: Shi, Liang
作者: Shi, Liang;Fang, Xing;Li, Mengjiao;Mu, Dashuai;Ren, Ang;Zhao, Mingwen;Tan, Qi
作者机构:
关键词: Ganoderma lucidum;Agrobacterium tumefaciens-mediated transformation;Promoter;Fluorescence assay;Histochemical staining assay
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: 2012 年 28 卷 1 期
页码:
收录情况: SCI
摘要: In this study, we report the development of a simple and efficient system for genetic transformation of the medicinal fungus Ganoderma lucidum. Various parameters were optimized to obtain successful Agrobacterium tumefaciens-mediated transformation. Co-cultivation of bacteria and protoplast at a ratio of 1,000:1 at 25A degrees C in medium containing 0.2 mM acetosyringone was found to be the optimum condition for high efficiency transformation. Four plasmids, each carrying a different promoter driving the expression of an antibiotic resistance marker, were tested. The construct carrying the Ganoderma lucidum glyceraldehyde-3-phosphate dehydrogenase (GPD) promoter showed good transformation efficiency, whereas constructs with the GPD promoter from ascomycetes were ineffective. Our analysis showed that over 70% of the transformants tested remained mitotically stable even after five successive rounds of subculturing. We were able to detect the expression of EGFP and GUS reporter genes in the Ganoderma lucidum transformants by fluorescence imaging and histochemical staining assays respectively. Our results demonstrate a new transgenic approach that will facilitate Ganoderma lucidum research.
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