Zhang Yongjun, Zhao Jianhua, Fang Weiguo, Zhang Jianqing, Luo Zhibing, Zhang Mi, Fan Yanhua, Pei Yan
Biotechnology Research Center, Southwest University, Chongqing, People's Republic of China.
Appl Environ Microbiol. 2009 Jun;75(11):3787-95. doi: 10.1128/AEM.01913-08. Epub 2009 Apr 10.
Beauveria bassiana is an economically important insect-pathogenic fungus which is widely used as a biocontrol agent to control a variety of insect pests. However, its insecticide efficacy in the field is often influenced by adverse environmental factors. Thus, understanding the genetic regulatory processes involved in the response to environmental stress would facilitate engineering and production of a more efficient biocontrol agent. Here, a mitogen-activated protein kinase (MAPK)-encoding gene, Bbhog1, was isolated from B. bassiana and shown to encode a functional homolog of yeast HIGH-OSMOLARITY GLYCEROL 1 (HOG1). A Bbhog1 null mutation was generated in B. bassiana by targeted gene replacement, and the resulting mutants were more sensitive to hyperosmotic stress, high temperature, and oxidative stress than the wild-type controls. These results demonstrate the conserved function of HOG1 MAPKs in the regulation of abiotic stress responses. Interestingly, DeltaBbhog1 mutants exhibited greatly reduced pathogenicity, most likely due to a decrease in spore viability, a reduced ability to attach to insect cuticle, and a reduction in appressorium formation. The transcript levels of two hydrophobin-encoding genes, hyd1 and hyd2, were dramatically decreased in a DeltaBbhog1 mutant, suggesting that Bbhog1 may regulate the expression of the gene associated with hydrophobicity or adherence.
球孢白僵菌是一种具有重要经济意义的昆虫病原真菌,被广泛用作生物防治剂来控制多种害虫。然而,其在田间的杀虫效果常受不利环境因素影响。因此,了解参与环境胁迫响应的基因调控过程将有助于构建和生产更高效的生物防治剂。在此,从球孢白僵菌中分离出一个编码丝裂原活化蛋白激酶(MAPK)的基因Bbhog1,并证明其编码酵母高渗甘油1(HOG1)的功能同源物。通过靶向基因替换在球孢白僵菌中产生了Bbhog1缺失突变体,与野生型对照相比,所得突变体对高渗胁迫、高温和氧化胁迫更敏感。这些结果证明了HOG1 MAPK在非生物胁迫响应调控中的保守功能。有趣的是,ΔBbhog1突变体的致病性显著降低,最可能的原因是孢子活力下降、附着于昆虫表皮的能力降低以及附着胞形成减少。在ΔBbhog1突变体中,两个编码疏水蛋白的基因hyd1和hyd2的转录水平显著降低,表明Bbhog1可能调控与疏水性或粘附相关基因的表达。