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Drought and Heat Stress-Mediated Modulation of Alternative Splicing in the Genes Involved in Biosynthesis of Metabolites Related to Tea Quality

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Abstract

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.

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Funding

This research was subsidized by the Significant Application Projects of Agriculture Technology Innovation in Shandong Province (SD2019ZZ010), the Technology System of Modern Agricultural Industry in Shandong Province (SDAIT-19-01) and the Special Foundation for Distinguished Taishan Scholar of Shangdong Province (no. ts201712057), the Livelihood Project of Qingdao City (19-6-1-64-nsh), the Project of Agricultural Science and Technology Fund in Shandong Province (2019LY002, 2019YQ010), the Key Research and Development Project of Shandong Province (2019LYXZ009).

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Authors and Affiliations

  1. Tea Research Institute, Shandong Academy of Agricultural Sciences, 276800, Rizhao, China

    Y. Q. Ding, K. Fan, Y. Wang, L. T. Sun, C. Qiu & Z. T. Ding

  2. College of Horticulture, Nanjing Agricultural University, 210095, Nanjing, China

    W. P. Fang & X. J. Zhu

  3. Tea Research Institute, Chinese Academy of Agricultural Sciences, 310008, Hangzhou, China

    L. Chen

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Contributions

Yiqian Ding and Kai Fan contributed equally to this article. Writing-Original Draft Preparation, Yiqian Ding; Writing-Review & Editing, Kai Fan and Zhaotang Ding; Methodology, Yu Wang and Liang Chen; Validation, Yiqian Ding and Litao Sun; Data Curation, Xujun Zhu; Software, Wanping Fang and Chen Qiu; Funding Acquisition, Zhaotang Ding.

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Abbreviations: ADH, alcohol dehydrogenase-like; AGXT2, alanine-glyoxylate aminotransferase 2; ALT, alanine transaminase; ANR, anthocyanidin reductase; argB, acetylglutamate kinase; CAD, cinnamyl alcohol dehydrogenase; CCD1, carotenoid cleavage dioxygenase 1; CHI, chalcone isomerase; DFR, dihydrofavonol-4-reductase; DFRA, dihydrofavonol-4-reductase-like; DXR, 1-deoxy-D-xylulose 5-phosphate reductoisomerase; DXS, probable 1-deoxy-D-xylulose-5-phosphate synthase; F3H, flavanone 3-hydroxylase; FLS, flavonol synthase; FPPS, farnesyl pyrophosphate synthase; GAD, glutamate decarboxylase; GDH, glutamate dehydrogenase; GGPPS1, geranylgeranyl diphosphate synthase 1; GOGAT1 (NADH), NADH-glutamate synthase; GS, glutamine synthetase; β‑GLU, beta-glucosidase; IDI, isopentenyl diphosphate isomerase; ilvE, branched-chain-amino-acid aminotransferase; leuA, 2-isopropylmalate synthase A-like; leuB, 3-isopropylmalate dehydrogenase; leuC, 3- isopropylmalate dehydrogenase large subunit; leuD, 3-isopropylmalate dehydrogenase small subunit; LIS, linalool synthase; LOX, lipoxygenase; lysC, aspartokinase; NADP-GDH, NADP-specific glutamate dehydrogenase; NAGS, amino-acid acetyltransferase; P5CDH, delta-1-pyrroline-5-carboxylate dehydrogenase; P5CS, pyrroline-5-carboxylate synthetase; PAL, phenylalanine ammonia-lyase; PMK, phosphomevalonate kinase; SHMT, serine hydroxy-methyltransferase; TES, (−)-alpha-terpineol synthase; thrA, bifunctional aspartokinase/homoserine dehydrogenase 1, chloroplast-like; thrC, threonine synthase, chloroplast-like; TPS, terpenoid/terpene synthase; VAT, acetolactate synthase small subunit.

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Ding, Y.Q., Fan, K., Wang, Y. et al. Drought and Heat Stress-Mediated Modulation of Alternative Splicing in the Genes Involved in Biosynthesis of Metabolites Related to Tea Quality. Mol Biol 56, 257–268 (2022). https://doi.org/10.1134/S0026893322020042

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