Cloning and Expression Analysis of AwDFRs in Himalayan Onion （Allium wallichii）

doi:10.11733/j.issn.1007-0435.2025.10.004

Acta Agrestia Sinica ›› 2025, Vol. 33 ›› Issue (10): 3165-3172.DOI: 10.11733/j.issn.1007-0435.2025.10.004

Cloning and Expression Analysis of AwDFRs in Himalayan Onion （Allium wallichii）

LIN Ying^1,3, TAN Yuan-yuan³, RAO Ying³, HUANG Qin², WANG Bo⁴, PENG Ting², LIU Guo-feng¹

1. Guangzhou Institute of Forestry and Landscape Architecture, Guangzhou, Guangdong Province 510405, China;
2. College of Life Science Technology, Huazhong Agricultural University, Wuhan, Hubei Province 430070, China;
3. College of Agriculture, Guizhou University, Guiyang, Guizhou Province 550025, China;
4. College of Plant Science Technology, Huazhong Agricultural University, Wuhan, Hubei Province 430070, China

Received:2024-12-18 Revised:2025-03-25 Published:2025-10-17

多星韭AwDFRs基因克隆及表达分析

林颖^1,3, 谭圆圆³, 饶英³, 黄覃², 汪波⁴, 彭婷², 刘国锋¹

1. 广州市林业和园林科学研究院, 广东广州 510405;
2. 华中农业大学生命科学技术学院, 湖北武汉 430070;
3. 贵州大学农学院, 贵州贵阳 550025;
4. 华中农业大学植物科学技术学院, 湖北武汉 430070

通讯作者: 刘国锋,E-mail:gfliu@mail.hzau.edu.cn;彭婷,E-mail:tpeng@mail.hzau.edu.cn
作者简介:刘国锋,E-mail:gfliu@mail.hzau.edu.cn;彭婷,E-mail:tpeng@mail.hzau.edu.cn
基金资助:
贵州省科技计划项目（黔科合基础-ZK[2022]一般095）；国家自然科学基金项目（32060365）；华中农业大学自主创新基金（2662022YJ019）资助

Abstract

Abstract: Dihydroflavonol 4-reductase （DFR） is a key enzyme in the anthocyanin synthesis pathway， and its expression is closely related to anthocyanin accumulation and flower colors. To explore the relationship between the DFR gene and flower colors in Allium wallichii， the purple and white flowers were used as materials， and DFR was cloned by RT-PCR and its expression pattern was analyzed. Four DFR allelic genes， called AwDFRs， were cloned， including two alleles AwDFR-P1 and AwDFR-P2 in the purple flowers， and two alleles AwDFR-W1 and AwDFR-W2 in the white flowers. Open reading frames （ORF） of all AwDFRs were 1155 bp， encoding 384 amino acids. Sequence alignment analysis revealed that the AwDFRs protein contained exactly the same NADPH conserved domains and substrate-specific binding domains. Phylogenetic tree analysis indicated that the AwDFRs were most closely related to onion DFR. The qRT-PCR results showed that expression of AwDFRs in petals was significantly higher than that in other tissues， and their expression increased with petal development and coloration in the pink， purple and deep purple flowers； However， AwDFRs were almost not expressed during the flower development stage in white flowers. The above results suggested that the different flower colors might be related to the differential expression of AwDFRs in A. wallichii. AwDFRs might play an important role in the anthocyanin synthesis and accumulation in A. wallichii.

Key words: Allium wallichii, Dihydroflavonol 4-reductase, Gene cloning, Expression analysis, Anthocyanins

摘要： 二氢黄酮醇4-还原酶（Dihydroflavonol 4-reductase，DFR）是花青素合成通路上的关键酶，其表达与花青素积累及花色呈现密切相关。为了探究DFR基因与多星韭（Allium wallichii）花色形成的关系，以紫花和白花多星韭为研究材料，利用RT-PCR技术克隆其DFR基因，并进行表达分析。结果克隆得到4条DFR等位基因，称作AwDFRs，包括紫花中的AwDFR-P1，AwDFR-P2和白花中的AwDFR-W1，AwDFR-W2；AwDFRs基因的开放阅读框均为1155 bp，编码384个氨基酸。序列比对分析表明，AwDFRs蛋白含有完全相同的NADPH保守结构域和底物特异性结合结构域。系统进化树分析表明，AwDFRs蛋白与洋葱DFR亲缘关系最近。qRT-PCR结果显示，AwDFRs在花瓣中的表达量显著高于其他组织，且在粉色、紫色和深紫色多星韭花中，随着花瓣发育和着色过程，AwDFRs的表达量逐渐升高；而在白花多星韭的整个花发育阶段，AwDFRs几乎都不表达。以上结果表明，多星韭花色差异形成的原因可能与AwDFRs基因的差异表达有关。AwDFRs在多星韭花青素合成积累过程中可能扮演着重要角色。

关键词: 多星韭, 二氢黄酮醇4-还原酶, 基因克隆, 表达分析, 花青素

CLC Number:

S682.1+9

LIN Ying, TAN Yuan-yuan, RAO Ying, HUANG Qin, WANG Bo, PENG Ting, LIU Guo-feng. Cloning and Expression Analysis of AwDFRs in Himalayan Onion （Allium wallichii）[J]. Acta Agrestia Sinica, 2025, 33(10): 3165-3172.

林颖, 谭圆圆, 饶英, 黄覃, 汪波, 彭婷, 刘国锋. 多星韭AwDFRs基因克隆及表达分析[J]. 草地学报, 2025, 33(10): 3165-3172.

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Cloning and Expression Analysis of AwDFRs in Himalayan Onion （Allium wallichii）

多星韭AwDFRs基因克隆及表达分析

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