意大利青霉菌对咪鲜胺的抗性机制 Click here to download the full text Citation：马一平,张保,付艳苹,朱福兴.意大利青霉菌对咪鲜胺的抗性机制.Journal of Plant Protection,2026,53(4):1041-1053 Hits: Download times: Author Name Affiliation E-mail Ma Yiping  College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China    Zhang Bao  College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China    Fu Yanping  College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China    Zhu Fuxing  College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China  zhufuxing@mail.hzau.edu.cn  中文摘要:为明晰意大利青霉菌Penicillium italicum对咪鲜胺的抗性机制，对采自四川、江西和湖北省的意大利青霉菌咪鲜胺抗性和敏感菌株的靶标酶基因CYP51A和CYP51B进行PCR扩增和测序，并测定其相对表达量，通过农杆菌介导的遗传转化和分子对接技术解析靶标酶基因CYP51A不同点突变对其抗性水平的影响。结果显示：意大利青霉菌咪鲜胺低抗菌株靶标酶CYP51B的4个氨基酸突变与高抗菌株的氨基酸突变完全一致，而对于靶标酶CYP51A，高抗菌株在低抗菌株突变的基础上另有F170L、A181T和F501L三个突变。通过遗传转化获得的单突变体D93E、F170L和A181T对咪鲜胺均保持敏感，而F501L突变体对咪鲜胺的抗性是出发菌株的20.2倍。携带D93E、F170L和A181T点突变的CYP51A蛋白与咪鲜胺的结合自由能较出发菌株的结合自由能无明显变化，而携带F501L突变的CYP51A蛋白与咪鲜胺的结合自由能由出发菌株的-8.99 kcal/mol升高为-8.68 kcal/mol。经咪鲜胺处理后，意大利青霉菌咪鲜胺低抗菌株靶标酶基因CYP51A的相对表达量是敏感菌株的2.29倍，高抗菌株CYP51B的相对表达量是敏感菌株的2.30倍。意大利青霉菌咪鲜胺低抗和高抗菌株的两个ABC转运蛋白基因的表达较敏感菌株均未升高。综上所述，靶标酶CYP51A中的F501L突变是意大利青霉菌对咪鲜胺产生高水平抗性的主要分子机制之一，靶标酶基因CYP51A和CYP51B的高表达可能在意大利青霉菌对咪鲜胺抗性中发挥一定作用。 中文关键词:意大利青霉菌  咪鲜胺  抗药性机制  CYP51A  点突变   Molecular mechanisms of prochloraz resistance in citrus blue mold pathogen Penicillium italicum Author Name Affiliation E-mail Ma Yiping  College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China    Zhang Bao  College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China    Fu Yanping  College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China    Zhu Fuxing  College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, Hubei Province, China  zhufuxing@mail.hzau.edu.cn  Abstract:To elucidate the resistance mechanisms of citrus blue mold pathogen Penicillium italicum to prochloraz, the target enzyme genes CYP51A and CYP51B were PCR-amplified and sequenced, and their relative expression levels were determined for prochloraz-resistant and sensitive isolates collected from Sichuan, Jiangxi, and Hubei provinces. Agrobacterium tumefaciens-mediated transformation (ATMT) and molecular docking were employed to investigate the effects of different mutations in the CYP51A gene on fungal resistance levels. Compared with sensitive isolates, the target enzyme CYP51B from low-resistance (LR) isolates harbored the same four-amino acid mutations as high-resistance (HR) isolates. However, for another target enzyme CYP51A, HR isolates had three additional mutations (F170L, A181T, and F501L) compared with LR isolates. ATMT demonstrated that mutants carrying the single-point mutation of D93E, F170L, or A181T remained sensitive to prochloraz, while the F501L mutant had a resistance ratio of 20.2 compared with the parent sensitive isolate. Molecular docking showed that the binding affinity of CYP51A to prochloraz for the D93E, F170L, and A181T mutants showed no significant difference compared with the sensitive isolates, whereas the binding free energy for the mutant F501L increased from -8.99 kcal/mol to -8.68 kcal/mol. After treatment with prochloraz, CYP51A expression in LR isolates was upregulated 2.29-fold, and CYP51B expression in HR isolates was upregulated 2.30-fold, relative to sensitive isolates. In contrast, the expression levels of two ATP-binding cassette (ABC) transporter genes were not significantly increased in either LR or HR isolates. In conclusion, the F501L mutation in CYP51A was the primary mechanism for high levels of prochloraz resistance in P. italicum, and elevated expression of the target genes CYP51A and CYP51B might also contribute to prochloraz resistance. keywords:Penicillium italicum  prochloraz  resistance mechanism  CYP51A  point mutation View Full Text  View/Add Comment  Download reader

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