The biosynthesis of EGCG, theanine and caffeine in response to temperature is mediated by hormone signal transduction factors in tea plant (Camellia sinensis L.)

Abstract

As the main flavor components of tea, the contents of epigallocatechin-3-gallate (EGCG), theanine and caffeine are regulated by ambient temperature. However, whether the biosynthesis of EGCG, theanine and caffeine in response to temperature is regulated by endogenous hormones and its mechanism is still unclear. In this study, tea cuttings cultivated in the phytotron which treated at different temperatures 15℃, 20℃, 25℃ and 30℃, respectively. The UPLC and ESI-HPLC-MS/MS were used to determine the contents of EGCG, theanine, caffeine and the contents of phytohormones in one leaf and a bud. The results showed that indoleacetic acid (IAA), gibberellin 1(GA1) and gibberellin 3 (GA3) were significantly correlated with the content of EGCG; Jasmonic acid (JA), jasmonate-isoleucine (JA-Ile) and methyl jasmonate (MeJA) were strongly correlated with theanine content; IAA, GA1 and gibberellin 4 (GA4) were significantly correlated with caffeine content at different temperatures. In order to explore the internal intricate relationships between the biosynthesis of these three main taste components, endogenous hormones, and structural genes in tea plants, we used multi-omics and multidimensional correlation analysis to speculate the regulatory mechanisms: IAA, GA1 and GA3 up-regulated the expressions of <i>chalcone synthase</i> (<i>CsCHS</i>) and <i>trans-cinnamate 4-monooxygenase</i> (<i>CsC4H</i>) mediated by the signal transduction factors <i>auxin-responsive protein IAA</i> (<i>CsIAA</i>) and <i>DELLA protein</i> (<i>CsDELLA</i>), respectively, which promoted the biosynthesis of EGCG; IAA, GA3 and GA1 up-regulated the expression of <i>CsCHS</i> and <i>anthocyanidin synthase</i> (<i>CsANS</i>) mediated by <i>CsIAA</i> and <i>CsDELLA</i>, respectively, via the transcription factor <i>WRKY DNA-binding protein</i> (<i>CsWRKY</i>), and promoted the biosynthesis of EGCG; JA, JA-Ile and MeJA jointly up-regulated the expression of <i>carbonic anhydrase</i> (<i>CsCA</i>) and down-regulated the expression of <i>glutamate decarboxylase</i> (<i>CsgadB</i>) mediated by the signal transduction factors <i>jasmonate ZIM domain-containing protein</i> (<i>CsJAZ</i>), and promoted the biosynthesis of theanine; JA, JA-Ile and MeJA also jointly inhibited the expression of <i>CsgadB</i> mediated by <i>CsJAZ</i> via the transcription factor <i>CsWRKY</i> and <i>AP2 family protein</i> (<i>CsAP2</i>), which promoted the biosynthesis of theanine; IAA inhibited the expression of <i>adenylosuccinate synthase</i> (<i>CspurA</i>) mediated by <i>CsIAA</i> via the transcription factor <i>CsWRKY</i>; GA1 and gibberellin 4 (GA4) inhibited the expression of <i>CspurA</i> mediated by <i>CsDELLA</i> through the transcription factor <i>CsWRKY</i>, which promoted the biosynthesis of caffeine. In conclusion, we revealed the underlying mechanism of the biosynthesis of the main taste components in tea plant in response to temperature was mediated by hormone signal transduction factors, which provided novel insights into improving the quality of tea.