Deeply functional identification of TCS1 alleles provides efficient technical paths for low-caffeine breeding of tea plants
文献类型： 外文期刊
作者： Wang, Yi 1 ; Liu, Yu-Fei 1 ; Wei, Meng-Yuan 1 ; Zhang, Chen-Yu 1 ; Chen, Jie-Dan 1 ; Yao, Ming-Zhe 1 ; Chen, Liang 1 ; Jin, Ji-Qiang 1 ;
作者机构： 1.Chinese Acad Agr Sci, Tea Res Inst, Minist Agr & Rural Affairs, Key Lab Biol Genet & Breeding Special Econ Anim &, Hangzhou 310008, Peoples R China
2.Yunnan Acad Agr Sci, Tea Res Inst, Yunnan Prov Key Lab Tea Sci, 2 Jingnan Rd, Menghai 666201, Yunnan, 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： 2023 年 10 卷 2 期
页码：
收录情况： SCI
摘要： Caffeine is an important functional component in tea, which has the effect of excitement and nerve stimulation, but excessive intake can cause insomnia and dysphoria. Therefore, the production of tea with low-caffeine content can meet the consumption needs of certain people. Here, in addition to the previous alleles of the tea caffeine synthase (TCS1) gene, a new allele (TCS1h) from tea germplasms was identified. Results of in vitro activity analysis showed that TCS1h had both theobromine synthase (TS) and caffeine synthase (CS) activities. Site-directed mutagenesis experiments of TCS1a, TCS1c, and TCS1h demonstrated that apart from the 225th amino acid residue, the 269th amino acid also determined the CS activity. GUS histochemical analysis and dual-luciferase assay indicated the low promoter activity of TCS1e and TCS1f. In parallel, insertion and deletion mutations in large fragments of alleles and experiments of site-directed mutagenesis identified a key cis-acting element (G-box). Furthermore, it was found that the contents of purine alkaloids were related to the expression of corresponding functional genes and alleles, and the absence or presence and level of gene expression determined the content of purine alkaloids in tea plants to a certain extent. In summary, we concluded TCS1 alleles into three types with different functions and proposed a strategy to effectively enhance low-caffeine tea germplasms in breeding practices. This research provided an applicable technical avenue for accelerating the cultivation of specific low-caffeine tea plants.
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