Functional characterization of cytochrome P450 CYP81A subfamily to disclose the pattern of cross-resistance in Echinochloa phyllopogon
Functional characterization of cytochrome P450 CYP81A subfamily to disclose the pattern of cross-resistance in Echinochloa phyllopogon
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DOI：
10.1007/s11103-019-00954-3
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发表时间：
2020-01-03
影响因子：
5.1
通讯作者：
Iwakami, Satoshi
中科院分区：
文献类型：
--
作者：
Dimaano, Nina Gracel;Yamaguchi, Takuya;Iwakami, Satoshi
关键信息CYP 81 A P450 s装甲Echinochloa phyllopogon对多种和几种除草剂化学品。CYP 81 A底物选择性可作为大肠杆菌交叉耐药性预测和控制的基础。phyllopogon和其他相关物种。基于代谢的除草剂抗性是对农业的一个主要威胁，因为它是不可预测的，并且可以扩展对不同化学基团和作用模式的抗性，包括现有的，新的和待发现的除草剂。有关除草剂代谢酶的信息有限，阻碍了杂草交叉抗性的预测。CYP 81 A亚家族成员在多重抗除草剂（MHR）的稗草phyllopogon先前被确定为赋予6个无关的除草剂类的交叉抗性。这表明CYP 81 A在赋予大肠杆菌不可预测的交叉耐药性中起着关键作用。因此，对该基因的9个功能基因对24个化学类群的33种除草剂的作用进行了研究。拟南芥中的异位表达鉴定了CYP 81 A，其可以赋予对多种和不同除草剂的抗性。CYP 81 A在大肠杆菌中的酶功能进一步表征。CYP 81 A在E.通过N-末端修饰、与HemA基因共表达、最适温度培养等方法，对原核表达载体进行了优化。CYP 81 A将其除草剂底物代谢成羟基化、N-/O-脱甲基化或两者兼而有之的产物。CYP 81 As赋予的交叉抗性模式针对乙酰乳酸合酶抑制剂的所有化学基团，并扩展到抑制光系统II、八氢番茄红素去饱和酶、原卟啉原氧化酶、4-羟苯基丙酮酸双加氧酶和1-脱氧-d-木酮糖5-磷酸合酶的除草剂。在MHR E中预测并证实了对除草剂嘧磺草醚、丙嗪磺隆和甲基磺草酮的交叉抗性。叶芒草本研究表明，除草剂代谢的关键酶的功能特性可以揭示交叉抗性模式，并确定适当的化学选择，以管理现有的和意外的交叉抗性在E.叶芒草
Key message CYP81A P450s armor Echinochloa phyllopogon against diverse and several herbicide chemistries. CYP81A substrate preferences can be a basis for cross-resistance prediction and management in E. phyllopogon and other related species. Metabolism-based herbicide resistance is a major threat to agriculture, as it is unpredictable and could extend resistance to different chemical groups and modes of action, encompassing existing, novel and to-be-discovered herbicides. Limited information on the enzymes involved in herbicide metabolism has hindered the prediction of cross-resistance in weeds. Members of CYP81A subfamily in multiple herbicide resistant (MHR) Echinochloa phyllopogon were previously identified for conferring cross-resistance to six unrelated herbicide classes. This suggests a critical role of CYP81As in endowing unpredictable cross-resistances in E. phyllopogon, thus the functions of all its nine putative functional CYP81A genes to 33 herbicides from 24 chemical groups were characterized. Ectopic expression in Arabidopsis thaliana identified the CYP81As that can confer resistance to multiple and diverse herbicides. The CYP81As were further characterized for their enzymatic functions in Escherichia coli. CYP81A expression in E. coli was optimized via modification of the N-terminus, co-expression with HemA gene and culture at optimal temperature. CYP81As metabolized its herbicide substrates into hydroxylated, N-/O-demethylated or both products. The cross-resistance pattern conferred by CYP81As is geared towards all chemical groups of acetolactate synthase inhibitors and is expanded to herbicides inhibiting photosystem II, phytoene desaturase, protoporphyrinogen oxidase, 4-hydroxyphenylpyruvate dioxygenase, and 1-deoxy-d-xylulose 5-phosphate synthase. Cross-resistance to herbicides pyrimisulfan, propyrisulfuron, and mesotrione was predicted and confirmed in MHR E. phyllopogon. This study demonstrated that the functional characterization of the key enzymes for herbicide metabolism could disclose the cross-resistance pattern and identify appropriate chemical options to manage the existing and unexpected cross-resistances in E. phyllopogon.