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Cloning and functional analysis of promoter of CsMIXTA associated with development of glandular trichome in industrial hemp

ZHOU Zhi-Man1,ZHANG Xiao-Yu1,GAO Feng2,DAI Zhi-Gang1,XU Ying1,CHENG Chao-Hua1,YANG Ze-Mao1,SU Jian-Guang1,TANG Qing1,*   

  1. 1 Institute of Bast Fiber Crops, Chinese Academy of Agricultural Sciences / Laboratory of Germplasm Resources, Changsha 410205, Hunan, China; 2 Yunnan Academy of Industrial Hemp, Kunming 650214, Yunnan, China
  • Received:2024-01-02 Revised:2024-06-20 Accepted:2024-06-20 Published:2024-07-12
  • Supported by:
    This study was supported by the China Agriculture Research System of MOF and MARA (Bast and Leaf Fiber Crops, CARS-16-E01) and the Science and Technology Innovation Project of the Chinese Academy of Agricultural Sciences (CAAS-ASTIP-2017-IBFC01). 

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Abstract:

CsMIXTA may play a crucial role in the morphology and development of glandular trichomes in cannabis female flowers. To investigate its regulatory mechanism, a 2199 bp promoter sequence of CsMIXTA was cloned. PlantCARE prediction identified multiple hormone response elements and stress response elements within this region. Based on this analysis, five 5′ end deletion fragments of the promoter with varying lengths were amplified. Six GUS gene expression vectors were constructed using the full-length promoter and the 5′ end deletion fragments, which were then transiently expressed in tobacco leaves and industrial hemp sugar leaves. GUS staining revealed that nucleotide positions -393 and -99 constituted the core region of the CsMIXTA promoter, containing the gibberellin response element TATC-box and the transcription initiation element TATA-box. The results also demonstrated that CsMIXTA is specifically expressed in glandular trichomes of industrial hemp. Promoter activity of the core region was confirmed by luciferase assay. Stress response analysis indicated that low temperature, abscisic acid (ABA), and gibberellin (GA3) enhanced promoter activity. These findings provide a crucial basis for further studies on the regulation of CsMIXTA.

Key words: industrial hemp, CsMIXTA promoter, transient expression, GA3 and ABA treatment

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