Molecular Biotechnology Laboratory of Triticeae Crops, Huazhong Agricultural University, Wuhan, PR China.
Fungal Genet Biol. 2010 Mar;47(3):205-15. doi: 10.1016/j.fgb.2009.11.003. Epub 2009 Nov 24.
Natural resistance of wheat against Fusarium head blight (FHB) is inadequate and new strategies for controlling the disease are required. Chitin synthases that catalyze chitin biosynthesis would be an ideal target for antifungal agents. In this study, a class I chitin synthase gene (CHS1) from Fusarium asiaticum, the predominant species of FHB pathogens on wheat in China, was functionally disrupted via Agrobacterium tumefaciens-mediated transformation. Specific disruption of the CHS1 gene resulted in a 58% reduction of chitin synthase activity, accompanied by decreases of 35% in chitin content, 22% in conidiation, and 16% in macroconidium length. The Deltachs1 mutant strain had a growth rate comparable to that of the wild-type on PDA medium but had a 35% increase in the number of nuclear cellulae and exhibited a remarkably increased sensitivity to osmosis stresses. Electron microscopy revealed substantial changes occurring in cell wall structures of the macroconidium, ascospore, and mycelium, with the most profound changes in the mycelium. Furthermore, the Deltachs1 mutant displayed significantly reduced pathogenicity on wheat spikes and seedlings. Re-introduction of a functional CHS1 gene into the Deltachs1 mutant strain restored the wild-type phenotype. These results reveal an important in vivo role played by a CHS1 gene in a FHB pathogen whose mycelial chitin could serve as a target for controlling the disease.
小麦对赤霉病(FHB)的天然抗性不足,因此需要新的策略来控制这种疾病。催化几丁质生物合成的几丁质合酶将是抗真菌剂的理想靶标。在这项研究中,通过根癌农杆菌介导的转化,功能破坏了来自中国小麦赤霉病主要病原菌亚洲镰刀菌的 I 类几丁质合酶基因(CHS1)。CHS1 基因的特异性破坏导致几丁质合酶活性降低 58%,同时几丁质含量降低 35%,产孢量减少 22%,大分生孢子长度减少 16%。与野生型相比,Deltachs1 突变菌株在 PDA 培养基上的生长速度相当,但核细胞数量增加了 35%,对渗透压胁迫的敏感性显著增加。电子显微镜显示,大分生孢子、子囊孢子和菌丝细胞壁结构发生了实质性变化,其中菌丝变化最为明显。此外,Deltachs1 突变体在小麦穗和幼苗上的致病性显著降低。将一个功能性 CHS1 基因重新引入 Deltachs1 突变菌株中,恢复了野生型表型。这些结果揭示了一个在体内发挥重要作用的 CHS1 基因在一种赤霉病病原体中的作用,其菌丝几丁质可以作为控制这种疾病的靶标。