College of Biological and Environmental Engineering, Zhejiang University of Technology, Hangzhou, 310032, People's Republic of China.
Curr Genet. 2013 Aug;59(3):119-28. doi: 10.1007/s00294-013-0393-y. Epub 2013 May 31.
Fungal chitin synthase of classes V and VI (or VII), which contain an additional N-terminal myosin motor domain, have been shown to play important roles in pathogenesis. To study the function of BcChsVI in Botrytis cinerea, BcChs6 gene was disrupted through Agrobacterium tumefaciens-mediated transformation. The Bcchs6 disruption mutant exhibited a 45.5 % increasing in its chitin content when compared with wild strain. The qRT-PCR analysis revealed that in Bcchs6 mutant the expression of BcChs6 was significantly decreased, while the expression of BcChs2 and BcChs3a was increased when compared with wild type. It is probable that the disruption of this gene provoked a compensatory mechanism regulating the cellular response to cell wall damage. Interestingly, the radial growth of Bcchs6 mutant was drastically reduced when 50 % solute was removed from the regular PDA medium, and they were more sensitive to Calcofluor white and other cell wall disturbing chemicals. Pathogenicity assays on tomato leaves indicated that they were significantly reduced in their ability to cause disease. Our results demonstrated that BcChs6 is necessary for proper hyphal growth and pathogenicity of B. cinerea on tomato leaves.
真菌的 V 类和 VI 类（或 VII 类）几丁质合酶，含有额外的 N 端肌球蛋白马达结构域，已被证明在发病机制中发挥重要作用。为了研究 BcChsVI 在 Botrytis cinerea 中的功能，通过农杆菌介导的转化破坏了 BcChs6 基因。与野生菌株相比，Bcchs6 缺失突变体的几丁质含量增加了 45.5%。qRT-PCR 分析显示，与野生型相比，Bcchs6 突变体中 BcChs6 的表达显著降低，而 BcChs2 和 BcChs3a 的表达增加。可能是该基因的破坏引发了一种细胞对细胞壁损伤的细胞反应的补偿机制。有趣的是，当从常规 PDA 培养基中去除 50%的溶质时，Bcchs6 突变体的径向生长急剧减少，它们对 Calcofluor white 和其他细胞壁破坏化学物质更加敏感。在番茄叶片上的致病性测定表明，它们引起疾病的能力显著降低。我们的结果表明，BcChs6 对于 B. cinerea 在番茄叶片上的菌丝生长和致病性是必要的。
