Highly-efficient liposome-mediated transformation system for the basidiomycetous fungus Flammulina velutipes

The Journal of General and Applied Microbiology

Ang Ren¹  Dongdong Chen¹  Mingwen Zhao¹  Hanshou Yu²  Liang Shi²  Chao Xu² 

[1] Key Laboratory of Microbiological Engineering of Agricultural Environment, Ministry of Agriculture;College of Life Sciences, Nanjing Agricultural University

关键词: Flammulina velutipes;    liposome-mediated transformation;    transformation efficiency;   

DOI  :  10.2323/jgam.2016.10.003

学科分类：微生物学和免疫学

来源: Applied Microbiology, Molecular and Cellulrar Biosciences Research Foundation

【 摘 要 】

Flammulina velutipes is a well-known edible mushroom cultivated all over the world. However, because of the low transformation frequency, the expensive instruments required, and the complicated, time-consuming procedures necessary, there is insufficient genetic research on F. velutipes. In this study, we report a liposome-mediated transformation (LMT) system for the genetic transformation of F. velutipes. Using the LMT system, we obtained 82 ± 4 stable F. velutipes transformants per 105 protoplasts, which is a clear increase in transformation frequency compared to the other methods used. We were able to detect the expression of an EGFP reporter gene in the F. velutipes transformants using fluorescence imaging assays. Furthermore, we used this method to transfer the laccase gene into F. velutipes and found that the transcriptional level and enzymatic activity increased in these transformants. Mitotic stability analysis showed that all of the selected transformants remained mitotically stable, even after five successive rounds of sub-culturing. These results demonstrate a new transgenic approach that will facilitate F. velutipes research.

【 授权许可】

Unknown