薏苡ClWRKY46基因的克隆、表达及抗旱功能研究

doi:10.3724/SP.J.1006.2026.64073

作物学报 ›› 2026, Vol. 52 ›› Issue (11): 3232-3247.doi: 10.3724/SP.J.1006.2026.64073

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

薏苡ClWRKY46基因的克隆、表达及抗旱功能研究

王玉娇^1,**，王永乐^1,**，路献勇¹，许大凤¹，付瑜华²，朱加保^1,*

1 安徽省农业科学院经济作物研究所, 安徽合肥 230001; 2 贵州省农业科学院亚热带作物研究所, 贵州贵阳 550025

收稿日期:2026-06-10 修回日期:2026-08-21 接受日期:2026-08-21 出版日期:2026-11-12 网络出版日期:2026-08-25
通讯作者: 朱加保, E-mail: 13955611798@139.com
基金资助:
本研究由国家自然科学基金青年科学基金项目(3240151950)和安徽省农业科学院自主创新项目(2026YL023)资助。

Cloning and expression of the ClWRKY46 gene and its function in drought resistance in Coix L.

Wang Yu-Jiao^1,**,Wang Yong-Le^1,**,Lu Xian-Yong¹,Xu Da-Feng¹,Fu Yu-Hua²,Zhu Jia-Bao^1,*

1 Institute of Industrial Crops, Anhui Academy of Agricultural Sciences, Hefei 230001, Anhui, China; 2 Guizhou Institute of Subtropical Crops, Guizhou Academy of Agricultural Sciences, Guiyang 550025, Guizhou, China

Received:2026-06-10 Revised:2026-08-21 Accepted:2026-08-21 Published:2026-11-12 Published online:2026-08-25
Supported by:
This study was supported by the Young Scientists Fund of the National Natural Science Foundation of China (3240151950), and the Independent Innovation Project of Anhui Academy of Agricultural Sciences (2026YL023).

摘要/Abstract

摘要： WRKY转录因子家族广泛参与植物干旱胁迫应答调控。薏苡作为一种重要的药食兼用杂粮作物，其抗旱分子机制研究相对薄弱。本研究旨在从薏苡中克隆ClWRKY46基因并解析其序列特征和表达模式，进而通过异源表达系统鉴定该基因在干旱胁迫下的应激响应与耐受功能，为薏苡耐旱遗传改良提供候选基因与理论依据。本研究采用RT-PCR技术克隆获得ClWRKY46基因，并分析其在不同组织中及干旱、盐等胁迫条件下的表达模式；通过亚细胞定位及酵母转录激活试验明确其编码蛋白的细胞定位与转录活性；同时利用酵母和拟南芥异源表达系统，对转基因酵母和转基因拟南芥进行抗旱性表型与生理指标鉴定。结果表明，ClWRKY46蛋白含有典型WRKY结构域，属于第III类WRKY亚家族，与水稻OsWRKY45同源性最高。该蛋白定位于细胞核中，且酵母转录激活试验证实其在酵母系统中无转录自激活活性。干旱胁迫可显著诱导ClWRKY46的转录上调。在酵母中异源表达ClWRKY46可显著提高菌株在干旱胁迫下的存活率。同时，转基因拟南芥在干旱胁迫下表现出更高的萌发率和更优的生长态势，其体内超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)等抗氧化酶活性显著升高，丙二醛(MDA)含量显著降低。综上所述，薏苡ClWRKY46是定位于细胞核的WRKY转录因子，无转录自激活活性，在植物响应干旱胁迫中发挥正向调控作用，异源表达证明该编码基因能够显著增强酵母和拟南芥的耐旱能力。

关键词: 薏苡, ClWRKY46, 转录激活, 亚细胞定位, 酵母功能验证, 非生物胁迫, 抗旱性

Abstract: The WRKY transcription factor family is widely involved in regulating plant responses to drought stress. Coix (Coix L.) is an important dual-purpose crop used for medicine and food, but its molecular mechanisms of drought tolerance have been relatively understudied. This study aimed to clone the ClWRKY46 gene from coix, analyze its sequence characteristics and expression patterns, and evaluate its stress response and tolerance under drought stress using heterologous expression systems, thereby providing a candidate gene and a theoretical basis for the genetic improvement of drought tolerance in coix. The ClWRKY46 gene was cloned using reverse transcription-polymerase chain reaction (RT-PCR), and its expression patterns in various tissues and under drought and salt stresses were analyzed. Protein localization and transcriptional activity were determined via subcellular localization and yeast transcriptional activation assays. Furthermore, drought tolerance phenotypes and physiological indicators were evaluated using yeast and Arabidopsis heterologous expression systems. The results showed that the cloned ClWRKY46 encodes a protein containing a typical WRKY domain, belonging to the Group Ⅲ WRKY subfamily, and sharing the highest homology with rice OsWRKY45. The ClWRKY46 protein is localized in the nucleus and lacks transcriptional autoactivation activity in the yeast system. Drought stress significantly induced the upregulation of ClWRKY46 expression. Heterologous expression of ClWRKY46 in yeast significantly improved the survival rate of the strain under drought stress. Meanwhile, transgenic Arabidopsis lines overexpressing ClWRKY46 exhibited higher germination rates and superior growth performance under drought conditions, accompanied by significantly increased activities of antioxidant enzymes (SOD, POD, CAT) and a significant reduction in malondialdehyde (MDA) content. In conclusion, ClWRKY46 is a nuclear-localized transcription factor lacking autoactivation activity, which plays a positive regulatory role in drought stress responses. Heterologous expression demonstrated that this gene can significantly enhance the drought resistance of yeast and Arabidopsis.

Key words: Coix lacryma-jobi L., ClWRKY46, transcriptional activation, subcellular localization, yeast functional verification, abiotic stress, drought resistance

王玉娇, 王永乐, 路献勇, 许大凤, 付瑜华, 朱加保. 薏苡ClWRKY46基因的克隆、表达及抗旱功能研究[J]. 作物学报, 2026, 52(11): 3232-3247.

Wang Yu-Jiao, Wang Yong-Le, Lu Xian-Yong, Xu Da-Feng, Fu Yu-Hua, Zhu Jia-Bao. Cloning and expression of the ClWRKY46 gene and its function in drought resistance in Coix L.[J]. Acta Agronomica Sinica, 2026, 52(11): 3232-3247.

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链接本文: https://zwxb.chinacrops.org/CN/10.3724/SP.J.1006.2026.64073

https://zwxb.chinacrops.org/CN/Y2026/V52/I11/3232

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薏苡ClWRKY46基因的克隆、表达及抗旱功能研究

Cloning and expression of the ClWRKY46 gene and its function in drought resistance in Coix L.

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