Cheng Xingkai, Dai Tan, Hu Zhihong, Cui Tongshan, Wang Weizhen, Han Ping, Hu Maolin, Hao Jianjun, Liu Pengfei, Liu Xili
Department of Plant Pathology, China Agricultural University, Beijing, China.
Institute of Quality Standard and Testing Technology, Beijing Academy of Agriculture and Forestry Sciences, Beijing, China.
Front Microbiol. 2022 Mar 21;13:806339. doi: 10.3389/fmicb.2022.806339. eCollection 2022.
SYP-14288 is a fungicide as an uncoupler of oxidative phosphorylation, which is effective in controlling fungal pathogens like To determine whether can develop SYP-14288 resistance and possibly multi-drug resistance (MDR), an SYP-14288-resistant mutant of X19-7 was generated from wild-type strain X19, and the mechanism of resistance was studied through metabolic and genetic assays. From metabolites of treated with SYP-14288, three compounds including M1, M2, and M3 were identified according to UPLC-MS/MS analysis, and M1 accumulated faster than M2 and M3 in X19-7. When X19-7 was treated by glutathione-S-transferase (GST) inhibitor diethyl maleate (DEM) and SYP-14288 together, or by DEM plus one of tested fungicides that have different modes of action, a synergistic activity of resistance occurred, implying that GSTs promoted metabolic resistance against SYP-14288 and therefore led to MDR. By comparing RNA sequences between X19-7 and X19, six cytochrome P450s (P450s) and two GST genes were selected as a target, which showed a higher expression in X19-7 than X19 both before and after the exposure to SYP-14288. Furthermore, heterologous expression of P450 and GST genes in yeast was conducted to confirm genes involved in metabolic resistance. In results, the P450 gene and GST gene were related to fungal resistance to multiple fungicides including SYP-14288, fluazinam, chlorothalonil, and difenoconazole. It was the first report that metabolic resistance of to uncouplers was associated with P450 and GST genes.
SYP - 14288是一种作为氧化磷酸化解偶联剂的杀菌剂，对诸如[具体真菌病原体名称未给出]等真菌病原体具有控制效果。为了确定[具体真菌名称未给出]是否会产生对SYP - 14288的抗性以及可能的多药耐药性（MDR），从野生型菌株X19中获得了X19 - 7的SYP - 14288抗性突变体，并通过代谢和遗传分析研究了抗性机制。根据超高效液相色谱 - 串联质谱（UPLC - MS/MS）分析，从经SYP - 14288处理的[具体真菌名称未给出]代谢产物中鉴定出三种化合物，包括M1、M2和M3，且在X19 - 7中M1的积累速度比M2和M3快。当X19 - 7同时用谷胱甘肽 - S - 转移酶（GST）抑制剂马来酸二乙酯（DEM）和SYP - 14288处理，或用DEM加一种具有不同作用模式的受试杀菌剂处理时，出现了抗性协同活性，这意味着GST促进了对SYP - 14288的代谢抗性，从而导致多药耐药性。通过比较X19 - 7和X19之间的RNA序列，选择了六个细胞色素P450（P450s）和两个GST基因作为靶点，这些靶点在暴露于SYP - 14288之前和之后在X19 - 7中的表达均高于X19。此外，在酵母中进行了P450和GST基因的异源表达以确认参与代谢抗性的基因。结果表明，P450基因[具体基因名称未给出]和GST基因[具体基因名称未给出]与真菌对包括SYP - 14288、氟啶胺、百菌清和苯醚甲环唑在内的多种杀菌剂的抗性有关。这是关于[具体真菌名称未给出]对解偶联剂的代谢抗性与P450和GST基因相关的首次报道。
