Ding Y Q, Fan K, Wang Y, Fang W P, Zhu X J, Chen L, Sun L T, Qiu C, Ding Z T
Tea Research Institute, Shandong Academy of Agricultural Sciences, Rizhao, 276800 China.
College of Horticulture, Nanjing Agricultural University, Nanjing, 210095 China.
Mol Biol (Mosk). 2022 Mar-Apr;56(2):321-322. doi: 10.31857/S0026898422020057.
Alternative splicing (AS) regulates mRNAs at the post-transcriptional level to affect both their amounts and the protein function. However, little is known about the roles of AS in regulation of biosynthesis of amino acids, flavonoids, and volatile compounds in tea plants. In this study, we used Iso-seq and transcriptome deep sequencing (RNA-seq) to identify AS events, and analyzed the expression of respective mRNAs in tea plants under drought (DS), heat stress (HS), and their combination (HD). By RT-PCR, we validated the AS events in nine genes involved in the biosynthesis of amino acids and flavonoids. The genes accumulating AS transcripts under DS, HS, and HD conditions included those encoding for anthocyanidin reductase (ANR), dihydrofavonol-4-reductase-like (DFRA), and chalcone isomerase (CHI). Similarly, genes directly or indirectly involved in the biosynthesis of volatile compounds such as lipoxygenase (LOX), terpenoid/terpene synthase (TPS), and 1-deoxy-D-xylulose 5-phosphate reductoisomerase (DXR) also had AS events. Our study revealed that AS might specifically regulate the biosynthesis of amino acids in tea plants under stressful conditions. Moreover, we suggest that the AS events within the ANR and DFRA transcripts might play an important role in the regulation of flavonoid biosynthesis under DS, HS, and HD conditions. This study improved our understanding of the genetic drivers of the changes in the content of bioactive ingredients of tea plants subjected to abiotic stresses.
可变剪接(AS)在转录后水平调控mRNA,从而影响其数量和蛋白质功能。然而,关于AS在茶树氨基酸、黄酮类化合物和挥发性化合物生物合成调控中的作用,我们知之甚少。在本研究中,我们利用全长转录组测序(Iso-seq)和转录组深度测序(RNA-seq)来鉴定AS事件,并分析了茶树在干旱(DS)、热胁迫(HS)及其组合(HD)条件下各mRNA的表达情况。通过逆转录聚合酶链反应(RT-PCR),我们验证了参与氨基酸和黄酮类化合物生物合成的9个基因中的AS事件。在DS、HS和HD条件下积累AS转录本的基因包括编码花青素还原酶(ANR)、类二氢黄酮醇-4-还原酶(DFRA)和查尔酮异构酶(CHI)的基因。同样,直接或间接参与挥发性化合物生物合成的基因,如脂氧合酶(LOX)、萜类/萜烯合酶(TPS)和1-脱氧-D-木酮糖-5-磷酸还原异构酶(DXR)也存在AS事件。我们的研究表明,AS可能在胁迫条件下特异性调控茶树氨基酸的生物合成。此外,我们认为ANR和DFRA转录本中的AS事件可能在DS、HS和HD条件下黄酮类化合物生物合成的调控中发挥重要作用。本研究增进了我们对遭受非生物胁迫的茶树生物活性成分含量变化的遗传驱动因素的理解。