College of Forestry, Sichuan Agricultural University, Chengdu 611130, China.
National Forestry and Grassland Administration Key Laboratory of Forest Resources Conservation and Ecological Safety on the Upper Reaches of the Yangtze River, Chengdu 611130, China.
Int J Mol Sci. 2022 Aug 31;23(17):9908. doi: 10.3390/ijms23179908.
can cause twig blight of the walnut (), resulting in great economic losses and ecological damage. We performed proteomic tandem mass tags (TMT) quantification of two strains with different substrates, BH01 in walnut substrate (SW) and sterile water (SK), and BH03 in walnut substrate (WW) and sterile water (WK), in order to identify differentially expressed proteins. We identified 998, 95, and 489 differentially expressed proteins (DEPs) between the SK vs. WK, SW vs. SK, and WW vs. WK comparison groups, respectively. A phylogenetic analysis was performed to classify the ABC transporter proteins annotated in the TMT protein quantification into eight groups. Physicochemical and structural analyses of the 24 ATP-binding cassette (ABC) transporter proteins revealed that 14 of them had transmembrane structures. To elucidate the functions of these transmembrane proteins, we determined the relative expression levels of ABC transporter genes in strains cultured in sodium chloride, hydrogen peroxide, copper sulfate, and carbendazim mediums, in comparison with pure medium; analysis revealed differential upregulation. To verify the expression results, we knocked out the gene and compared the wild-type and knockout mutant strains. The knockout mutant strains exhibited a higher sensitivity to antifungal drugs. Furthermore, the virulence of the knockout mutant strains was significantly lower than the wild-type strains, thus implying that plays a role in the drug resistance of and affects its virulence.
可引起核桃枝枯病(),造成巨大的经济损失和生态破坏。我们对在核桃基质(SW)和无菌水(SK)中生长的 菌株 BH01 和在核桃基质(WW)和无菌水(WK)中生长的 菌株 BH03 这两种具有不同基质的菌株进行了串联质量标签(TMT)定量蛋白质组学分析,以鉴定差异表达蛋白。我们分别在 SK 与 WK、SW 与 SK 和 WW 与 WK 比较组中鉴定到 998、95 和 489 个差异表达蛋白(DEPs)。我们对 TMT 蛋白定量注释的 ABC 转运蛋白进行了系统发育分析,将其分为 8 个组。对 24 个 ATP 结合盒(ABC)转运蛋白的理化性质和结构分析表明,其中 14 个具有跨膜结构。为了阐明这些跨膜蛋白的功能,我们测定了在氯化钠、过氧化氢、硫酸铜和多菌灵培养基中培养的菌株中 ABC 转运基因的相对表达水平,与纯培养基相比,发现这些基因发生了差异上调。为了验证表达结果,我们敲除了 基因,并比较了野生型和敲除突变菌株。敲除突变菌株对抗真菌药物的敏感性更高。此外,敲除突变菌株的毒力明显低于野生型菌株,这表明 基因在 菌株的耐药性中发挥作用,并影响其毒力。