Key Laboratory of Tea Quality and Safety Control, Ministry of Agriculture, Tea Research Institute, Chinese Academy of Agricultural Sciences, No. 9 Meiling South Road, Xihu District, Hangzhou, 310008, Zhejiang, China.
BMC Plant Biol. 2021 Nov 9;21(1):521. doi: 10.1186/s12870-021-03254-5.
Shoot branching is one of the important agronomic traits affecting yields and quality of tea plant (Camellia sinensis). Cytokinins (CTKs) play critical roles in regulating shoot branching. However, whether and how differently alternative splicing (AS) variant of CTKs-related genes can influence shoot branching of tea plant is still not fully elucidated.
In this study, five AS variants of CTK biosynthetic gene adenylate isopentenyltransferase (CsA-IPT5) with different 3' untranslated region (3' UTR) and 5' UTR from tea plant were cloned and investigated for their regulatory effects. Transient expression assays showed that there were significant negative correlations between CsA-IPT5 protein expression, mRNA expression of CsA-IPT5 AS variants and the number of ATTTA motifs, respectively. Shoot branching processes induced by exogenous 6-BA or pruning were studied, where CsA-IPT5 was demonstrated to regulate protein synthesis of CsA-IPT5, as well as the biosynthesis of trans-zeatin (tZ)- and isopentenyladenine (iP)-CTKs, through transcriptionally changing ratios of its five AS variants in these processes. Furthermore, the 3' UTR AS variant 2 (3AS2) might act as the predominant AS transcript.
Together, our results indicate that 3AS2 of the CsA-IPT5 gene is potential in regulating shoot branching of tea plant and provides a gene resource for improving the plant-type of woody plants.
分枝是影响茶树(Camellia sinensis)产量和品质的重要农艺性状之一。细胞分裂素(CTKs)在调控分枝中起着关键作用。然而,CTKs 相关基因的可变剪接(AS)变体是否以及如何影响茶树的分枝还没有完全阐明。
本研究从茶树中克隆了 5 种具有不同 3'非翻译区(3'UTR)和 5'UTR 的 CTK 生物合成基因腺苷酸异戊烯基转移酶(CsA-IPT5)的 AS 变体,并研究了它们的调控作用。瞬时表达分析表明,CsA-IPT5 蛋白表达、CsA-IPT5 AS 变体的 mRNA 表达与 ATTTA 基序的数量之间存在显著的负相关。通过外源 6-BA 或修剪诱导的分枝过程研究表明,CsA-IPT5 通过转录改变其 5 种 AS 变体的比例,调节 CsA-IPT5 的蛋白合成以及反式玉米素(tZ)和异戊烯腺嘌呤(iP)-CTKs 的生物合成,从而调控分枝过程。此外,3'UTR AS 变体 2(3AS2)可能作为主要的 AS 转录本发挥作用。
综上所述,我们的研究结果表明,CsA-IPT5 基因的 3AS2 可能参与调控茶树的分枝,为改良木本植物的株型提供了基因资源。