Resistance to the novel fungicide pyrimorph in Phytophthora capsici: risk assessment and detection of point mutations in CesA3 that confer resistance.
Resistance to the novel fungicide pyrimorph in Phytophthora capsici: risk assessment and detection of point mutations in CesA3 that confer resistance.
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辣椒疫霉对新型杀菌剂吡吗啉的抗性：风险评估和 CesA3 中赋予抗性的点突变的检测
DOI：
10.1371/journal.pone.0056513
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Liu X
中科院分区：
文献类型：
--
作者：
Pang Z;Shao J;Chen L;Lu X;Hu J;Qin Z;Liu X
吡吗啉是一种新型杀菌剂，对辣椒疫霉菌具有很高的抑制活性。我们研究了辣椒疫霉对丁酰吗啉产生抗性的风险。226株辣椒疫霉的基线敏感性，通过菌丝生长抑制测试，显示单峰分布，平均EC 50值为1.4261（±0.4002）µg/ml。通过在体外以约1×10−4的频率重复暴露于嘧啶吗啉，获得了12个嘧啶吗啉抗性突变体。突变体的抗性系数在10.67 ~ 56.02之间。突变体在无嘧菌灵培养基上转10次后，对嘧菌灵的抗性稳定。健身在孢子形成，孢子萌发，和致病性的梨形抗突变体是相似的或小于在亲本野生型菌株。然而，在用推荐的最大剂量的丁酰吗啉处理的离体辣椒叶片和辣椒植株上，具有高水平丁酰吗啉抗性的突变体的毒力大于野生型。结果表明，辣椒疫霉菌对丁酰吗啉产生抗性的风险为低至中等。在具有高水平的啶虫吗啉抗性的突变体中，啶虫吗啉和CAA杀菌剂氟吗啉、烯酰吗啉和双炔酰菌胺的EC 50值呈正相关。这表明纤维素合成酶3（CesA 3）中的点突变可赋予对丁酰吗啉的抗性。CesA 3在具有高水平的丁酰吗啉抗性的分离株和亲本分离株中的比较表明，在位置1077处从谷氨酰胺到赖氨酸的氨基酸变化导致突变体中稳定的高抗性。基于点突变，建立了检测辣椒疫霉菌对丁酰吗啉抗性的等位基因特异性PCR方法。
Pyrimorph is a novel fungicide with high activity against the plant pathogen Phytophthora capsici. We investigated the risk that P. capsici can develop resistance to pyrimorph. The baseline sensitivities of 226 P. capsici isolates, tested by mycelial growth inhibition, showed a unimodal distribution with a mean EC50 value of 1.4261 (±0.4002) µg/ml. Twelve pyrimorph-resistant mutants were obtained by repeated exposure to pyrimorph in vitro with a frequency of approximately 1×10−4. The resistance factors of the mutants ranged from 10.67 to 56.02. Pyrimorph resistance of the mutants was stable after 10 transfers on pyrimorph-free medium. Fitness in sporulation, cystospore germination, and pathogenicity in the pyrimorph-resistant mutants was similar to or less than that in the parental wild-type isolates. On detached pepper leaves and pepper plants treated with the recommended maximum dose of pyrimorph, however, virulence was greater for mutants with a high level of pyrimorph resistance than for the wild type. The results suggest that the risk of P. capsici developing resistance to pyrimorph is low to moderate. Among mutants with a high level of pyrimorph resistance, EC50 values for pyrimorph and CAA fungicides flumorph, dimethomorph, and mandipropamid were positively correlated. This indicated that point mutations in cellulose synthase 3 (CesA3) may confer resistance to pyrimorph. Comparison of CesA3 in isolates with a high level of pyrimorph resistance and parental isolates showed that an amino acid change from glutamine to lysine at position 1077 resulted in stable, high resistance in the mutants. Based on the point mutations, an allele-specific PCR method was developed to detect pyrimorph resistance in P. capsici populations.
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