转录组学分析揭示BcatrB介导灰葡萄孢对嘧霉胺的抗性

BcatrB mediates pyrimethanil resistance in Botrytis cinerea revealed by transcriptomics analysis.

作者信息

Wang Shuqi, Zhang Xinyi, Zhong Shan, Wang Rong, Chen Bingwei, Ding Wanlong, Luo Hongmei, Li Yong

机构信息

State Key Laboratory for Quality Ensurance and Sustainable Use of Dao-di Herbs, Chinese Academy of Medical Sciences, Peking Union Medical College, Institute of Medicinal Plant Development, Beijing, 100193, People's Republic of China.

出版信息

Sci Rep. 2025 Apr 3;15(1):11478. doi: 10.1038/s41598-025-96041-3.

DOI:10.1038/s41598-025-96041-3

PMID:40181028

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11968863/

Abstract

Gray mold, caused by Botrytis cinerea, poses an escalating threat to ginseng production in China. Excessive application of chemical fungicides has resulted in severe resistance development. To elucidate the resistance mechanism of B. cinerea to pyrimethanil, transcriptomic comparisons were conducted between resistant (HRG21) and sensitive (FSG43) strains following exposure to pyrimethanil for 2 and 6 h. GO and KEGG analyses identified differential expression of genes associated with ABC and MFS transporters as well as methionine biosynthesis. qRT-PCR validation confirmed a marked upregulation of ABC (BcatrA, BcatrB, BcatrD, BcatrO, and Bmr3) and MFS (Bchex1 and BcmfsM2) transporter genes in HRG21, whereas in FSG43, ABC (BcatrA, BcatrB, BcatrD, and BcatrO) and MFS (Bchex1) transporter genes were downregulated. No significant transcriptional changes were observed in Mrr1, a transcription factor gene, following pyrimethanil exposure. Molecular docking analysis identified potential binding sites and interaction targets for five key transporters (BcatrB, BcatrA, BcatrD, Bmr3, and Bchex1). Furthermore, exogenous methionine supplementation partially mitigated pyrimethanil toxicity in FSG43. However, amino acid sequences of enzymes involved in methionine biosynthesis (BcmetC, BcStr2, Bcmet2, Bcmxr1, and Bcmxr2) exhibited no variation between FSG43 and HRG21. The results indicate that BcatrB plays a central role in pyrimethanil resistance, while methionine contributes only marginally to resistance mechanisms.

摘要

由灰葡萄孢引起的灰霉病对中国的人参生产构成了日益严重的威胁。化学杀菌剂的过度使用导致了严重的抗药性问题。为了阐明灰葡萄孢对嘧霉胺的抗性机制，在将抗性菌株（HRG21）和敏感菌株（FSG43）暴露于嘧霉胺2小时和6小时后，进行了转录组比较。基因本体（GO）和京都基因与基因组百科全书（KEGG）分析确定了与ABC和MFS转运蛋白以及蛋氨酸生物合成相关的基因差异表达。实时定量聚合酶链反应（qRT-PCR）验证证实，HRG21中ABC（BcatrA、BcatrB、BcatrD、BcatrO和Bmr3）和MFS（Bchex1和BcmfsM2）转运蛋白基因显著上调，而在FSG43中，ABC（BcatrA、BcatrB、BcatrD和BcatrO）和MFS（Bchex1）转运蛋白基因下调。在暴露于嘧霉胺后，转录因子基因Mrr1未观察到明显的转录变化。分子对接分析确定了五个关键转运蛋白（BcatrB、BcatrA、BcatrD、Bmr3和Bchex1）的潜在结合位点和相互作用靶点。此外，外源补充蛋氨酸可部分减轻FSG43中嘧霉胺的毒性。然而，FSG43和HRG21之间参与蛋氨酸生物合成的酶（BcmetC、BcStr2、Bcmet2、Bcmxr1和Bcmxr2）的氨基酸序列没有差异。结果表明，BcatrB在嘧霉胺抗性中起核心作用，而蛋氨酸对抗性机制的贡献很小。