Purple-leaf tea plants, as anthocyanin-rich cultivars, are valuable materials for manufacturing teas with unique colors or flavors. In this study, a new purple-leaf cultivar “Zixin” (“ZX”) was examined, and its biochemical variation and mechanism of leaf color change were elucidated. The metabolomes of leaves of “ZX” at completely purple, intermediately purple, and completely green stages were analyzed using ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF-MS). Metabolites in the flavonoid biosynthetic pathway remained at high levels in purple leaves, whereas intermediates of porphyrin and chlorophyll metabolism and carotenoid biosynthesis exhibited high levels in green leaves. In addition, fatty acid metabolism was more active in purple leaves, and steroids maintained higher levels in green leaves. Saponin, alcohol, organic acid, and terpenoid-related metabolites also changed significantly during the leaf color change process. Furthermore, the substance changes between “ZX” and “Zijuan” (a thoroughly studied purple-leaf cultivar) were also compared. The leaf color change in “Zijuan” was mainly caused by a decrease in flavonoids/anthocyanins. However, a decrease in flavonoids/anthocyanins, an enhancement of porphyrin, chlorophyll metabolism, carotenoid biosynthesis, and steroids, and a decrease in fatty acids synergistically caused the leaf color change in “ZX”. These findings will facilitate comprehensive research on the regulatory mechanisms of leaf color change in purple-leaf tea cultivars.

Tea [Camellia sinensis (L.) O. Kuntze] is an important economic crop, and drought is the most important abiotic stress affecting yield and quality. Abscisic acid (ABA) is an important phytohormone responsible for activating drought resistance. Increased understanding of ABA effects on tea plant under drought stress is essential to develop drought-tolerant tea genotypes, along with crop management practices that can mitigate drought stress. The objective of the present investigation is evaluation of effects of exogenous ABA on the leaf proteome in tea plant exposed to drought stress. Leaf protein patterns of tea plants under simulated drought stress [(polyethylene glycol (PEG)-treated] and exogenous ABA treatment were analyzed in a time-course experiment using two-dimensional electrophoresis (2-DE), followed by matrix-assisted laser desorption/ionization time-of-flight (MALDI-TOF) mass spectrometry (MS). Among the 72 protein spots identified by MALDI-TOF MS, 16 proteins were downregulated and two were upregulated by exogenous ABA. The upregulated proteins have roles in glycolysis and photosystem II stabilization. Twenty-one protein spots were responsive to drought stress and most participate in carbohydrate and nitrogen metabolism, control of reactive oxygen species (ROS), defense, signaling or nucleic acid metabolism. The combined treatments of exogenous ABA and drought showed upregulation of 10 protein spots at 12 h and upregulation of 11 proteins at 72 h after initiation of drought stress. The results support the importance of the role that ABA plays in the tea plant during drought stress, by improving protein transport, carbon metabolism and expression of resistance proteins.

To determine the mechanisms in tea plants responding to temperature stresses (heat and cold), we examined the global transcriptomic and metabolomic profiles of the tea plant cultivar ‘Suchazao’ under moderately low temperature stress (ML), severely low temperature stress (SL), moderately high temperature stress (MH) and severely high temperature stress (SH) using RNA-seq and high performance liquid chromatography tandem mass spectrometry/mass spectrometry (HPLC-MS/MS), respectively. The identified differentially expressed genes indicated that the synthesis of stress-resistance protein might be redirected to cope with the temperature stresses. We found that heat shock protein genes Hsp90 and Hsp70 played more critical roles in tea plants in adapting to thermal stress than cold, while late embryogenesis abundant protein genes (LEA) played a greater role under cold than heat stress, more types of zinc finger genes were induced under cold stress as well. In addition, energy metabolisms were inhibited by SH, SL and ML. Furthermore, the mechanisms of anthocyanin synthesis were different under the cold and heat stresses. Indeed, the CsUGT75C1 gene, encoding UDP-glucose:anthocyanin 5-O-glucosyl transferase, was up-regulated in the SL-treated leaves but down-regulated in SH. Metabolomics analysis also showed that anthocyanin monomer levels increased under SL. These results indicate that the tea plants share certain foundational mechanisms to adjust to both cold and heat stresses. They also developed some specific mechanisms for surviving the cold or heat stresses. Our study provides effective information about the different mechanisms tea plants employ in surviving cold and heat stresses, as well as the different mechanisms of anthocyanin synthesis, which could speed up the genetic breeding of heat- and cold-tolerant tea varieties.

Low-temperature stress is an increasing problem for the cultivation of tea (Camellia sinensis), with adverse effects on plant growth and development and subsequent negative impacts on the tea industry. Methyl jasmonate (MeJA), as a plant inducer, can improve the cold-stress tolerance in tea plants. R2R3-MYB transcription factors (TFs) are considered potentially important regulators in the resistance to cold stress in plants. However, the molecular mechanisms, by which MYB TFs via the jasmonic acid pathway respond to cold stress in the tea plant, remain unknown. In this study, physiological and biochemical assays showed that exogenous MeJA application could effectively promote ROS scavenging in the tea plant under cold stress, maintaining the stability of the cell membrane. Sixteen R2R3-MYB TFs genes were identified from the tea plant genome database. Quantitative RT-PCR analysis showed that three CsMYB genes were strongly induced under a combination of MeJA and cold-stress treatment. Subcellular localization assays suggest CsMYB45, CsMYB46, and CsMYB105 localized in the nucleus. Exogenous MeJA treatment enhanced the overexpression of CsMYB45, CsMYB46, and CsMYB105 in E. coli and improved the growth and survival rates of recombinant cells compared to an empty vector under cold stress. Yeast two-hybrid and bimolecular fluorescence complementation experiments confirmed that CsMYB46 and CsMYB105 interacted with CsJAZ3, CsJAZ10, and CsJAZ11 in the nucleus. Taken together, these results highlight that CsMYB45, CsMYB46, and CsMYB105 are not only key components in the cold-stress signal response pathway but also may serve as points of confluence for cold stress and JA signaling pathways. Furthermore, our findings provide new insight into how MYB TFs influence cold tolerance via the jasmonic acid pathway in tea and provide candidate genes for future functional studies and breeding.