Functional characterization of NADPH-cytochrome P450 reductase and cinnamic acid 4-hydroxylase encoding genes from Scoparia dulcis L.

IF 4.1 3区生物学 Q1 PLANT SCIENCES

Botanical Studies Pub Date : 2020-03-02 DOI:10.1186/s40529-020-00284-4

Yoshimi Yamamura, Ayaka Mabuchi

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引用次数: 3

Abstract

Background: Most plant cytochrome P450 (P450) proteins need to be supplied with electrons from a redox partner, e.g. an NADPH-cytochrome P450 reductase (CPR), for the activation of oxygen molecules via heme. CPR is a flavoprotein with an N-terminal transmembrane domain, which transfers electrons from NADPH to the P450 via coenzymes flavin adenine dinucleotide and flavin mononucleotide.

Results: In this study, a novel CPR (SdCPR) was isolated from a tropical medicinal plant Scoparia dulcis L. The deduced amino acid of SdCPR showed high homology of > 76% with CPR from higher plants and belonged to the class II CPRs of dicots. Recombinant SdCPR protein reduced cytochrome c, ferricyanide (K₃Fe(CN)₆), and dichlorophenolindophenol in an NADPH-dependent manner. To elucidate the P450 monooxygenase activity of SdCPR, we isolated a cinnamic acid 4-hydroxylase (SdC4H, CYP73A111) gene from S. dulcis. Biochemical characterization of SdCPR/SdC4H demonstrated that SdCPR supports the oxidation step of SdC4H. Real-time qPCR results showed that expression levels of SdCPR and SdC4H were inducible by mechanical wounding treatment and phytohormone elicitation (methyl jasmonate, salicylic acid), which were consistent with the results of promotor analyses.

Conclusions: Our results showed that the SdCPR and SdC4H are related to defense reactions, including the biosynthesis of secondary metabolites.

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茯苓nadph -细胞色素P450还原酶和肉桂酸4-羟化酶编码基因的功能研究。

背景:大多数植物细胞色素P450 (P450)蛋白需要由氧化还原伙伴提供电子，例如nadph -细胞色素P450还原酶(CPR)，通过血红素激活氧分子。CPR是一种具有n端跨膜结构域的黄素蛋白，它通过辅酶黄素腺嘌呤二核苷酸和黄素单核苷酸将电子从NADPH转移到P450。结果:从热带药用植物东Scoparia dulcis L.中分离到一种新的CPR (SdCPR)，其氨基酸与高等植物的CPR同源性> 76%，属于薯蓣属II类CPR。重组SdCPR蛋白以nadph依赖的方式还原细胞色素c、铁氰化物(K3Fe(CN)6)和二氯酚吲哚酚。为了研究肉桂酸羟化酶的P450单加氧酶活性，我们从肉桂酸中分离出一个肉桂酸4-羟化酶(SdC4H, CYP73A111)基因。SdCPR/SdC4H的生化表征表明，SdCPR支持SdC4H的氧化步骤。Real-time qPCR结果显示，机械损伤处理和植物激素(茉莉酸甲酯、水杨酸)诱导均可诱导SdCPR和SdC4H的表达水平，这与启动子分析结果一致。结论:我们的研究结果表明SdCPR和SdC4H与防御反应有关，包括次生代谢物的生物合成。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Botanical Studies PLANT SCIENCES-

CiteScore

4.80

自引率

2.90%

发文量

审稿时长

13 weeks

期刊介绍： Botanical Studies is an open access journal that encompasses all aspects of botany, including but not limited to taxonomy, morphology, development, genetics, evolution, reproduction, systematics, and biodiversity of all plant groups, algae, and fungi. The journal is affiliated with the Institute of Plant and Microbial Biology, Academia Sinica, Taiwan.