School of Life Sciences, Chongqing University, Chongqing, 401331, People's Republic of China.
Chongqing Engineering Research Center for Fungal Insecticides, Chongqing, 401331, People's Republic of China.
Appl Microbiol Biotechnol. 2017 Jun;101(12):5033-5043. doi: 10.1007/s00253-017-8290-9. Epub 2017 Apr 19.
In fungi, the Ca/calcineurin signaling pathway is critical in mediating growth, morphology, stress responses, and pathogenicity. Crz1 is a calcineurin-responsive zinc finger transcription factor. Here, MaCrz1 was identified and functionally characterized in the entomopathogenic fungus Metarhizium acridum by characterization of a targeted gene knockout strain. Conidia of the ΔMacrz1 mutant were aberrant in cell surface features and lacked the characteristic hydrophobic rodlet layer, and the mutant displayed increased sensitivity to oxidative stress, cell wall perturbing agents, heat stress, and ultraviolet irradiation as compared to the wild-type and complemented strains. Insect bioassay using locusts revealed decreased virulence for the ΔMaCrz1 mutant, with defects in the ability of the mutant to penetrate the host cuticle. The ΔMaCrz1 mutant also showed greatly reduced chitin and β-1,3-glucan level in the cell wall. Transcriptomic profiling revealed genes involved in cell wall synthesis, conidiation, stress tolerance, and calcium transport that were downregulated in the ΔMaCrz1 mutant. Our results demonstrate that MaCrz1 plays important roles in stress susceptibility and pathogenicity, and provides clues as to the genes and pathways targeted by the transcription factor.
在真菌中,Ca/calcineurin 信号通路在介导生长、形态、应激反应和致病性方面起着至关重要的作用。Crz1 是一种钙调神经磷酸酶反应性锌指转录因子。在这里,通过靶向基因敲除菌株的表征,鉴定并功能表征了昆虫病原真菌绿僵菌中的 MaCrz1。与野生型和互补菌株相比,ΔMacrz1 突变体的分生孢子在细胞表面特征上异常,缺乏特征性的疏水性棒状层,并且对氧化应激、细胞壁破坏剂、热应激和紫外线照射的敏感性增加。使用蝗虫进行的昆虫生物测定显示,ΔMaCrz1 突变体的毒力降低,突变体穿透宿主表皮的能力受损。ΔMaCrz1 突变体在细胞壁中的几丁质和β-1,3-葡聚糖水平也大大降低。转录组分析显示,参与细胞壁合成、分生孢子形成、应激耐受和钙转运的基因在ΔMaCrz1 突变体中下调。我们的结果表明,MaCrz1 在应激易感性和致病性方面发挥着重要作用,并为转录因子靶向的基因和途径提供了线索。