RESUMEN
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 15â, 20â, 25â and 30â, 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 chalcone synthase (CsCHS) and trans-cinnamate 4-monooxygenase (CsC4H) mediated by the signal transduction factors auxin-responsive protein IAA (CsIAA) and DELLA protein (CsDELLA), respectively, which promoted the biosynthesis of EGCG; IAA, GA3 and GA1 up-regulated the expression of CsCHS and anthocyanidin synthase (CsANS) mediated by CsIAA and CsDELLA, respectively, via the transcription factor WRKY DNA-binding protein (CsWRKY), and promoted the biosynthesis of EGCG; JA, JA-Ile and MeJA jointly up-regulated the expression of carbonic anhydrase (CsCA) and down-regulated the expression of glutamate decarboxylase (CsgadB) mediated by the signal transduction factors jasmonate ZIM domain-containing protein (CsJAZ), and promoted the biosynthesis of theanine; JA, JA-Ile and MeJA also jointly inhibited the expression of CsgadB mediated by CsJAZ via the transcription factor CsWRKY and AP2 family protein (CsAP2), which promoted the biosynthesis of theanine; IAA inhibited the expression of adenylosuccinate synthase (CspurA) mediated by CsIAA via the transcription factor CsWRKY; GA1 and gibberellin 4 (GA4) inhibited the expression of CspurA mediated by CsDELLA through the transcription factor CsWRKY, 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.
RESUMEN
Epigallocatechin-3-gallate (EGCG), a flavoured and healthy compounds in tea, is affected by the ecological factors. However, the biosynthetic mechanisms of EGCG in response to the ecological factors remian unclear. In this study, a response surface method with a Box-Behnken design was used to investigate the relationship between EGCG accumulation and ecological factors; further, integrative transcriptome and metabolome analyses were performed to explore the mechanism underlying EGCG biosynthesis in response to environmental factors. The optimal environmental conditions obtained for EGCG biosynthesis were as follows: 28â, 70 % relative humidity of the substrate, and 280 µmol·m-2·s-1 light intensity; the EGCG content was increased by 86.83 % compared to the control (CK1). Meanwhile, the order of EGCG content in response to the interaction of ecological factors was as follows: interaction of temperature and light intensity > interaction of temperature and relative humidity of the substrate > interaction of light intensity and relative humidity of the substrate, indicating that temperature was the dominant ecological factors. EGCG biosynthesis in tea plants was found to be comprehensively regulated by a series of structural genes (CsANS, CsF3H, CsCHI, CsCHS, and CsaroDE), miRNAs (miR164, miR396d, miR5264, miR166a, miR171d, miR529, miR396a, miR169, miR7814, miR3444b, and miR5240), and transcription factors (MYB93, NAC2, NAC6, NAC43, WRK24, bHLH30, and WRK70); further, the metabolic flux was regulated and converted from phenolic acid to the flavonoid biosynthesis pathway based on accelerated consumption of phosphoenolpyruvic acid, d-erythrose-4-phosphate, and l-phenylalanine in response to ambient changes in temperature and light intensity. Overall, the results of this study reveal the effect of ecological factors on EGCG biosynthesis in tea plants, providing novel insights for improving tea quality.
Asunto(s)
Camellia sinensis , Camellia sinensis/química , Transcriptoma , Metaboloma , Té/químicaRESUMEN
BACKGROUND: Catechins are crucial in determining the flavour and health benefits of tea, but it remains unclear that how the light intensity regulates catechins biosynthesis. Therefore, we cultivated tea plants in a phytotron to elucidate the response mechanism of catechins biosynthesis to light intensity changes. RESULTS: In the 250 µmol·m- 2·s- 1 treatment, the contents of epigallocatechin, epigallocatechin gallate and total catechins were increased by 98.94, 14.5 and 13.0% respectively, compared with those in the 550 µmol·m- 2·s- 1 treatment. Meanwhile, the photosynthetic capacity was enhanced in the 250 µmol·m- 2·s- 1 treatment, including the electron transport rate, net photosynthetic rate, transpiration rate and expression of related genes (such as CspsbA, CspsbB, CspsbC, CspsbD, CsPsbR and CsGLK1). In contrast, the extremely low or high light intensity decreased the catechins accumulation and photosynthetic capacity of the tea plants. The comprehensive analysis revealed that the response of catechins biosynthesis to the light intensity was mediated by the photosynthetic capacity of the tea plants. Appropriately high light upregulated the expression of genes related to photosynthetic capacity to improve the net photosynthetic rate (Pn), transpiration rate (Tr), and electron transfer rate (ETR), which enhanced the contents of substrates for non-esterified catechins biosynthesis (such as EGC). Meanwhile, these photosynthetic capacity-related genes and gallic acid (GA) biosynthesis-related genes (CsaroB, CsaroDE1, CsaroDE2 and CsaroDE3) co-regulated the response of GA accumulation to light intensity. Eventually, the epigallocatechin gallate content was enhanced by the increased contents of its precursors (EGC and GA) and the upregulation of the CsSCPL gene. CONCLUSIONS: In this study, the catechin content and photosynthetic capacity of tea plants increased under appropriately high light intensities (250 µmol·m- 2·s- 1 and 350 µmol·m- 2·s- 1) but decreased under extremely low or high light intensities (150 µmol·m- 2·s- 1 or 550 µmol·m- 2·s- 1). We found that the control of catechin accumulation by light intensity in tea plants is mediated by the plant photosynthetic capacity. The research provided useful information for improving catechins content and its light-intensity regulation mechanism in tea plant.