Wang Gangzheng, Luo Yi, Wang Chen, Zhou Yan, Mou Chunye, Kang Heng, Xiao Yang, Bian Yinbing, Gong Yu Hua
Institute of Applied Mycology, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan, China.
State Key Laboratory of Applied Microbiology Southern China, Guangdong Provincial Key Laboratory of Microbial Culture Collection and Application, Guangdong Open Laboratory of Applied Microbiology, Guangdong Institute of Microbiology, Guangzhou, China.
Front Microbiol. 2020 Apr 17;11:707. doi: 10.3389/fmicb.2020.00707. eCollection 2020.
Our previous study found that RNAi decreased resistance to heat stress and infection. In this study, the structure and function of the gene was analyzed by gene cloning and overexpression in stress-sensitive strain YS55 via the -mediated transformation method. Transformants were confirmed by qRT-PCR, fluorescence observation and Southern blotting. Overexpression of in YS55 not only enhanced mycelial resistance to heat stress but also facilitated mycelial growth. In the presence of heat stress, the intracellular IAA content showed a significant increase in the two overexpression strains but only a slight change in the YS55 wild type strain. Moreover, the interaction between LeDnaJ07 and LetrpE was demonstrated via Y2H and BiFC assays. These results suggested that LeDnaJ07 may be involved in regulating IAA biosynthesis and the resistance of to heat stresses via interacting with LetrpE.
我们之前的研究发现，RNA干扰降低了对热应激和感染的抗性。在本研究中，通过基因克隆以及利用介导转化方法在应激敏感菌株YS55中进行过表达，对该基因的结构和功能进行了分析。通过实时定量聚合酶链反应、荧光观察和Southern印迹法对转化体进行了确认。在YS55中过表达不仅增强了菌丝体对热应激的抗性，还促进了菌丝体的生长。在热应激条件下，两个过表达菌株的细胞内吲哚-3-乙酸（IAA）含量显著增加，而YS55野生型菌株中只有轻微变化。此外，通过酵母双杂交（Y2H）和双分子荧光互补（BiFC）分析证实了LeDnaJ07与LetrpE之间的相互作用。这些结果表明，LeDnaJ07可能通过与LetrpE相互作用参与调节IAA生物合成以及对热应激的抗性。
