利用 CRISPR-Cas9 在大豆疫霉菌中证实了 PsORP1 基因的多个点突变赋予了对唑菌酯不同的抗性水平。
Multiple point mutations in PsORP1 gene conferring different resistance levels to oxathiapiprolin confirmed using CRISPR-Cas9 in Phytophthora sojae.
机构信息
State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling, China.
Department of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing, China.
出版信息
Pest Manag Sci. 2020 Jul;76(7):2434-2440. doi: 10.1002/ps.5784. Epub 2020 Feb 24.
BACKGROUND
Oxathiapiprolin is among the first commercial oxysterol-binding protein inhibitors (OSBPIs) developed by DuPont Corporation and shows excellent activity against plant-pathogenic oomycetes. Although more than 21 target site mutations have been identified in insensitive oomycetes, only G770V, G839W, and ΔN837 have been verified to confer oxathiapiprolin resistance in Phytophthora capsici or P. sojae. The effect of other mutations on OSBPIs sensitivity requires urgent investigation.
RESULTS
P. sojae transformants containing 16 mutations of PsORP1 were recovered using the CRISPR-Cas9 system. Transformants containing L733W, S768F, S768Y, N837Y, N837F, P861H, L863W, or I877Y showed high oxathiapiprolin resistance, with resistant factors (RFs) > 3000. Point mutations S768K, S768I, G770L, G770P, G770A, ΔG818/F819, N837I, and I877F exhibited low resistance, with RFs < 80. Phenotype assays revealed that the most highly resistant transformants showed enhanced or similar pathogenicity, oospore production, and cyst gemination. However, most transformants displayed decreased sporangia and zoospore production compared with parental wild-type P6497.
CONCLUSION
This study demonstrated that L733W, S768F, S768Y, N837Y, N837F, P861H, L863W, and I877Y in PsORP1 confer high oxathiapiprolin resistance in P. sojae.
背景
恶二唑啉是杜邦公司开发的首批商业甾醇结合蛋白抑制剂（OSBPIs）之一，对植物病原卵菌表现出优异的活性。虽然已经在不敏感的卵菌中鉴定出超过 21 个靶标位点突变，但只有 G770V、G839W 和 ΔN837 被证实能够赋予辣椒疫霉或大豆疫霉对恶二唑啉的抗性。其他突变对 OSBPIs 敏感性的影响需要紧急调查。
结果
使用 CRISPR-Cas9 系统回收了含有 16 种 PsORP1 突变的 P. sojae 转化体。含有 L733W、S768F、S768Y、N837Y、N837F、P861H、L863W 或 I877Y 的转化体表现出对恶二唑啉的高抗性，抗性因子（RF）>3000。点突变 S768K、S768I、G770L、G770P、G770A、ΔG818/F819、N837I 和 I877F 表现出低抗性，RFs<80。表型测定表明，最具抗性的转化体表现出增强或相似的致病性、卵孢子产生和胞囊萌发。然而，与亲本野生型 P6497 相比，大多数转化体的游动孢子和泡囊产生减少。
结论
本研究表明，PsORP1 中的 L733W、S768F、S768Y、N837Y、N837F、P861H、L863W 和 I877Y 赋予 P. sojae 对恶二唑啉的高抗性。
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