mRNA-miRNA analyses reveal the involvement of CsbHLH1 and miR1446a in the regulation of caffeine biosynthesis in Camellia sinensis
文献类型： 外文期刊
作者： Jin, Qifang 1 ; Wang, Zhong 1 ; Sandhu, Devinder 4 ; Chen, Lan 1 ; Shao, Chenyu 1 ; Shang, Fanghuizi 1 ; Xie, Siyi 1 ; Huang, Feiyi 5 ; Chen, Zhenyan 1 ; Zhang, Xiangqin 1 ; Hu, Jinyu 1 ; Liu, Guizhi 1 ; Su, Qin 1 ; Huang, Mengdi 1 ; Liu, Zhonghua 1 ; Huang, Jianan 1 ; Tian, Na 1 ; Liu, Shuoqian 1 ;
作者机构： 1.Hunan Agr Univ, Key Lab Tea Sci, Minist Educ, Changsha 410128, Peoples R China
2.Hunan Agr Univ, Natl Res Ctr Engn & Technol Utilizat Bot Funct Ing, Changsha 410128, Peoples R China
3.Hunan Agr Univ, CoInnovat Ctr Educ Minist Utilizat Bot Funct Ingre, Changsha 410128, Peoples R China
4.USDA, US Salin Lab, Agr Res Serv, Riverside, CA 92504 USA
5.Hunan Acad Agr Sci Natl Small & Medium Leaf Tea Pl, Tea Res Inst, Changsha 410128, Peoples R China; Hunan Agr Univ, Changsha 410128, Hunan, Peoples R China
期刊名称：HORTICULTURE RESEARCH （ 影响因子：8.7； 五年影响因子：9.0 ）
ISSN： 2662-6810
年卷期： 2024 年 11 卷 2 期
页码：
收录情况： SCI
摘要： Caffeine, a primary flavor component in tea, has been the subject of intense research. With the goal of shedding light on the complex regulatory processes governing caffeine biosynthesis in tea plants, liquid chromatography coupled with mass spectrometry (LC-MS), transcriptomics, and small RNA analyses were employed on diverse tea cultivars such as 'Jianghua Kucha' [including 'Xianghong 3' (XH3H) and 'Kucha 3' (KC3H)], 'Fuding Dabaicha' (FDDB), 'Yaoshan Xiulv' (YSXL), and 'Bixiangzao' (BXZ). The results showed that the caffeine level in 'Jianghua Kucha' was significantly higher than that in other tea plant cultivars. In addition, weighted gene co-expression network analysis indicated that that the CsbHLH1 gene might play a pivotal role as a potential hub gene related to the regulation of caffeine biosynthesis. Subcellular localization analysis showed that the CsbHLH1 protein was localized in the nucleus of the cells. Moreover, CsbHLH1 suppresses the transcription of TCS1 by binding to the TCS1 promoter, as evidenced by a yeast one-hybrid assay, an electrophoretic mobility shift assay, and dual luciferase analysis. In addition, a microRNA, miR1446a, was identified that directly cleaves CsbHLH1, leading to an increase in caffeine levels. Therefore, our findings imply that CsbHLH1 binds to the TCS1 promoter (-971 to -1019 bp) to reduce its expression, thereby negatively regulating caffeine biosynthesis. On the other hand, miR1446a enhances the biosynthesis of caffeine by suppressing the expression of CsbHLH1. This work enhances our understanding of the molecular mechanisms of caffeine biosynthesis in tea plants and offers potential directions for manipulating caffeine levels in future tea cultivation.
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