Functional genetic analysis of the leucinostatin biosynthesis transcription regulator lcsL in Purpureocillium lilacinum using CRISPR-Cas9 technology
Functional genetic analysis of the leucinostatin biosynthesis transcription regulator lcsL in Purpureocillium lilacinum using CRISPR-Cas9 technology
复制标题
利用 CRISPR-Cas9 技术对紫紫紫霉中亮氨抑素生物合成转录调节因子 lcsL 进行功能遗传分析
DOI：
10.1007/s00253-019-09945-2
复制
发表时间：
2019-08-01
影响因子：
5
通讯作者：
Xie, Bingyan
中科院分区：
文献类型：
--
作者：
Jiao, Yang;Li, Yan;Xie, Bingyan
淡紫紫青霉是一种很有前途的防治植物寄生线虫和植物病原菌的药剂。Leucinostatins是由淡紫青霉产生的脂肽家族，由20个基因组成的基因簇合成、修饰和调控。序列分析表明，lcsL，在lcs簇的基因，是一个假定的bZIP转录因子。在这项研究中，引入了CRISPR-Cas9系统来提高同源重组的效率，以破坏lcsL。在lcsL破坏突变体中，簇中基因的表达显著降低，并且leucinostatins的输出降低到检测不到的水平。在lcsL过表达菌株中，簇中基因的表达和leucinostatins的产量都增加。野生型和突变体对致病疫霉的拮抗作用也与基因表达和leucinostatins的产量一致。这些结果表明，基因lcsL是至关重要的调节leucinostatins的合成。
Purpureocillium lilacinum is a promising commercial agent for controlling plant-parasitic nematodes and plant pathogens. Leucinostatins are a family of lipopeptides produced by P. lilacinum that are synthesized, modified, and regulated by a gene cluster consisting of 20 genes. Sequence analyses have indicated that lcsL, a gene in the lcs cluster, is a putative bZIP transcription factor. In this study, the CRISPR-Cas9 system was introduced to increase the efficiency of homologous recombination for the disruption of lcsL. The expression of genes in the cluster was significantly reduced in lcsL disruption mutants, and the output of leucinostatins was decreased to undetectable levels. In the lcsL overexpression strain, the expression of genes in the cluster and the yield of leucinostatins were all increased. The antagonism of both the wild type and mutant against Phytophthora infestans was also consistent with the gene expression and the output of leucinostatins. These results indicate that the gene lcsL is crucial for the regulating the synthesis of leucinostatins.