Cloning and Functional Characterization of <italic style="font-style: italic">O</italic>-Methyltransferase Gene in <italic style="font-style: italic">Carthamus tinctorius</italic>

HUANG Yongming; LI Yaping; SU Ping; XIA Meng

doi:10.13422/j.cnki.syfjx.20260311

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Cloning and Functional Characterization of O-Methyltransferase Gene in Carthamus tinctorius

更新时间：2026-04-01

- Cloning and Functional Characterization of O-Methyltransferase Gene in Carthamus tinctorius
  增强出版
- Chinese Journal of Experimental Traditional Medical Formulae Vol. 32, Issue 9, Pages: 217-223(2026)
- 作者机构：
  
  1.中国中医科学院中药资源中心道地药材品质保障与资源持续利用全国重点实验室，北京 100700
  2.西南大学药学院，重庆 400715
- 作者简介：
- 基金信息：
- DOI：10.13422/j.cnki.syfjx.20260311
  CLC： R284.1;R286.0;R927
- Received：11 December 2025，
  
  Revised：2026-02-11，
  
  Accepted：25 February 2026，
  
  Online First：02 March 2026，
  
  Published：05 May 2026
- 稿件说明：
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黄永明,李娅萍,苏平等.红花O-甲基转移酶基因克隆与功能分析[J].中国实验方剂学杂志,2026,32(09):217-223.

HUANG Yongming,LI Yaping,SU Ping,et al.Cloning and Functional Characterization of O-Methyltransferase Gene in Carthamus tinctorius[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(09):217-223.
黄永明,李娅萍,苏平等.红花O-甲基转移酶基因克隆与功能分析[J].中国实验方剂学杂志,2026,32(09):217-223. DOI： 10.13422/j.cnki.syfjx.20260311.

HUANG Yongming,LI Yaping,SU Ping,et al.Cloning and Functional Characterization of O-Methyltransferase Gene in Carthamus tinctorius[J].Chinese Journal of Experimental Traditional Medical Formulae,2026,32(09):217-223. DOI： 10.13422/j.cnki.syfjx.20260311.

摘要

目的

对红花中的

-甲基转移酶基因进行全面鉴定与分析，并从中挖掘可催化黄酮类化合物甲基化的关键

-甲基转移酶基因，为解析红花中黄酮类化合物结构多样性的分子形成机制提供依据。

方法

基于课题组前期获取的红花高质量基因组数据，利用隐马尔可夫模型对红花Ⅰ型

-甲基转移酶基因进行系统鉴定。利用多种生物信息学在线工具，对鉴定得到的基因进行理化性质、基因结构、保守基序、染色体定位、基因复制事件及共线性分析。进一步通过大肠埃希菌原核表达系统对目标基因进行蛋白异源表达，并利用体外酶促反应验证其功能。

结果

共筛选出31个Ⅰ型红花

-甲基转移酶基因。从中成功克隆到一条红花

CtFOMT

1甲基转移酶基因，其编码蛋白在大肠埃希菌中实现可溶性表达，并通过Ni²⁺亲和色谱纯化获得高浓度蛋白。体外酶促试验结果表明，CtFOMT1能以

-腺苷甲硫氨酸为甲基供体，催化柚皮素4′-OH甲基化生成异樱花素；催化木犀草素4′-OH甲基化生成香叶木素，并进一步催化香叶木素3′-OH甲基化生成4′-甲基金圣草素。

结论

CtFOMT

1可催化黄酮类化合物骨架中4′-/3′-OH甲基化，推断其可能参与红花中多种4′-/3′-氧甲基黄酮类化合物生物合成。

Abstract

Objective

To comprehensively identify the

-methyltransferase （OMT） genes in

Carthamus tinctorius

and explore the key OMTs that can catalyze the methylation of flavonoids， providing a basis for understanding the molecular formation mechanism of the structural diversity of flavonoids in

C. tinctorius

Methods

The hidden Markov model was used to systematically identify the type Ⅰ OMTs from the high-quality genome data of

C. tinctorius

. A suite of bioinformatics tools was employed to systematically analyze the physicochemical properties， gene structure， conserved motifs， chromosomal localization， gene replication events， and collinearity of the identified genes. The target gene was heterologously expressed through the prokaryotic expression system of

E. coli

， and the protein function was verified by

in vitro

enzymatic reactions.

Results

A total of 31 type Ⅰ OMTs were identified.

CtFOMT

1 was successfully cloned and expressed in a soluble form in

Escherichia coli

. The recombinant protein was purified via Ni

affinity chromatography to obtain a high-concentration preparation.

In vitro

enzymatic assays demonstrated that CtFOMT1 utilized

-adenosylmethionine as the methyl donor to catalyze the methylation of the 4′-OH of naringenin， resulting in the production of isosakuranetin. A similar process occurred with the 4′-OH of luteolin， leading to the formation of diosmetin. Subsequent methylation of the 3′-OH group of diosmetin generated 4′-methylchrysoeriol.

Conclusion

CtFOMT

1 can catalyze the methylation of 4′-/3′-OH in the flavonoid skeleton. It is hypothesized that

CtFOMT

1 may play a role in the biosynthesis of various 4′-/3′-oxymethyl flavonoids in

C. tinctorius

关键词

Keywords

references

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Cloning and Functional Characterization of O-Methyltransferase Gene in Carthamus tinctorius

DOI：10.13422/j.cnki.syfjx.20260311

摘要

Abstract

关键词

Keywords

references