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

Wang Yu-Jiao1,**,Wang Yong-Le1,**,Lu Xian-Yong1,Xu Da-Feng1,Fu Yu-Hua2,Zhu Jia-Bao1,*   

  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:2026-08-25
  • 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).

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

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