Evolution of Benzimidazole Resistance Caused by Multiple Double Mutations of β-Tubulin in Corynespora cassiicola,Journal of Agricultural and Food Chemistry

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Evolution of Benzimidazole Resistance Caused by Multiple Double Mutations of β-Tubulin in Corynespora cassiicola
Journal of Agricultural and Food Chemistry ( IF 5.7 ) Pub Date : 2022-11-28 , DOI: 10.1021/acs.jafc.2c05912
Yushuai Mao ₁ , Haoran Li ₁ , Wen Song ₁ , Baoquan Zhao ₁ , Yiqiang Cai _{1,

2} , Jianxin Wang _{1,

2} , Mingguo Zhou _{1,

2} , Yabing Duan _{1,

2}

Affiliation

Cucumber target leaf spot caused by Corynespora cassiicola has devastated greenhouse cucumber production. In our previous study, the resistance monitoring of C. cassiicola to carbendazim was carried out, and a large number of resistant populations carrying various mutations (M163I&E198A, F167Y&E198A, F200S&E198A, or E198A) in β-tubulin were detected. However, the single-point mutations M163I, F167Y, and F200S have remained undetected. To investigate the evolutionary mechanism of double mutations in β-tubulin of C. cassiicola resistance to benzimidazoles, site-directed mutagenesis was used to construct alleles with corresponding mutation genotypes in β-tubulin. Through PEG-mediated protoplast transformation, all the mutants except for the M163I mutation were obtained and conferred resistance to benzimidazoles. It was found that the mutants conferring the E198A or double-point mutations showed high resistance to carbendazim and benomyl, but the mutants conferring the F167Y or F200S mutations showed moderate resistance. Except, the F200S mutants showed low resistance, the resistance level of the other mutants to thiabendazole seemed no difference. In addition, compared to the other mutants, the F167Y and F200S mutants suffered a more severe fitness penalty in mycelial growth, sporulation, and virulence. Thus, combined with the resistance level, fitness, and molecular docking results, we concluded that the field double mutations (F167Y&E198A and F200S&E198A) evolved from the single mutations F167Y and F200S, respectively.

中文翻译：

Corynespora cassiicola β-微管蛋白多重双突变引起的苯并咪唑耐药性的演变

由Corynespora cassiicola引起的黄瓜目标叶斑病破坏了温室黄瓜生产。我们在前期研究中对C. cassiicola对多菌灵的耐药性进行了监测，检测到大量携带β-tubulin各种突变（M163I&E198A、F167Y&E198A、F200S&E198A或E198A）的耐药群体。然而，单点突变 M163I、F167Y 和 F200S 仍未被发现。为了研究C. cassiicola对苯并咪唑的抗性β-微管蛋白双突变的进化机制，采用定点诱变构建了具有相应突变基因型的β-微管蛋白等位基因。. 通过 PEG 介导的原生质体转化，获得了除 M163I 突变外的所有突变体，并赋予了对苯并咪唑的抗性。结果发现，赋予E198A或双点突变的突变体对多菌灵和苯菌灵表现出高抗性，而赋予F167Y或F200S突变的突变体表现出中等抗性。除F200S突变体表现出低耐药性外，其余突变体对噻苯达唑的耐药水平似乎没有差异。此外，与其他突变体相比，F167Y 和 F200S 突变体在菌丝体生长、孢子形成和毒力方面遭受更严重的适应性损失。因此，结合抗性水平、适应度和分子对接结果，我们得出结论，田间双突变（F167Y&E198A和F200S&

更新日期：2022-11-28

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