蛹虫草
突变体
生物
转化(遗传学)
清脆的
分子生物学
生物化学
基因
植物
冬虫夏草
作者
Hong Sang Choi,S.J Park,Jeill Oh,Cheol Soo Kim,Gi‐Ho Sung,Hyunkyu Sang
出处
期刊:Fems Microbiology Letters
[Oxford University Press]
日期:2023-01-01
卷期号:370
标识
DOI:10.1093/femsle/fnad072
摘要
Abstract Cordyceps militaris, an entomopathogenic ascomycete, produces edible medicinal mushrooms known to have medicinal and therapeutic functions. To develop the genetic transformation system in C. militaris, green fluorescent protein (GFP) mutants of C. militaris were generated by PEG-mediated protoplast transformation. The CRISPR/Cas9 ribonucleoprotein (RNP) targeting the class III histidine kinase of C. militaris (CmHk1) was then delivered into protoplasts of C. militaris through the transformation system. Mutations induced by the RNP in selected mutants were detected: 1 nt deletion (6 mutants), 3 nt deletion with substitution of 1 nt (1 mutant), insertion of 85 nts (1 mutant), 41 nts (2 mutants), and 35 nts (5 mutants). An in vitro sensitivity assay of the mutants indicated that knockout of CmHk1 reduced sensitivity to two fungicides, iprodione and fludioxonil, but increased sensitivity to osmotic stresses compared to the wild type. Summing up, the CRISPR/Cas9 RNP delivery system was successfully developed, and our results revealed that CmHk1 was involved in the fungicide resistance and osmotic stress in C. militaris.
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