Rougui Wuyi Rock tea (WRT) with special flavor can be affected by multiple factors that are closely related to the culturing regions of tea plants. The present research adopted non-targeted metabolomics based on liquid chromatography–mass spectrometry (LC-MS) and gas chromatography–mass spectrometry (GC-MS), aroma activity value method (OAV), and chemometrics to analyze the characteristic metabolites of three Rougui WRTs from different culturing regions. The results of sensory evaluation showed that the three Rougui Wuyi Rock teas had significantly different flavor qualities, especially in taste and aroma. Rougui (RG) had a heavy and mellow taste, while cinnamon-like odor Rougui (GPRG) and floral and fruity odor Rougui (HGRG) had a thick, sweet, and fresh taste. The cinnamon-like odor was more obvious and persistent in GPRG than in RG and HGRG. HGRG had floral and fruity characteristics such as clean and lasting, gentle, and heavy, which was more obvious than in RG and GPRG. The results of principal component analysis (PCA) showed that there were significant metabolic differences among the three Rougui WRTs. According to the projection value of variable importance (VIP) of the partial least squares discriminant analysis (PLS–DA), 24 differential non-volatile metabolites were identified. The PLSR analysis results showed that rutin, silibinin, arginine, lysine, dihydrocapsaicin, etc. may be the characteristic non-volatiles that form the different taste outlines of Rougui WRT. A total of 90 volatiles, including aldehydes, alcohols, esters, and hydrocarbons, were identified from the three flavors of Rougui WRT by using GC-MS. Based on OAV values and PLS-DA analysis, a total of 16 characteristic volatiles were identified. The PLSR analysis results showed that 1-penten-3-ol, α-pinene, 2-carene, β-Pinene, dehydrolinalool, adipaldehyde, D-limonene, saffron aldehyde, and 6-methyl-5-hepten-2-one may be the characteristic volatiles that form the different aroma profile of Rougui WRT. These results provide the theoretical basis for understanding the characteristic metabolites that contribute to the distinctive flavors of Rougui WRT.

Tea aroma is a key indicator for evaluating tea quality. Although notable success in tea aroma improvement has been achieved with heterosis breeding technology, the molecular basis underlying heterosis remains largely unexplored. Thus, the present report studies the tea plant volatile heterosis using a high-throughput next-generation RNA-seq strategy and gas chromatography–mass spectrometry. Phenotypically, we found higher terpenoid volatile and green leaf volatile contents by gas chromatography–mass spectrometry in the F1 hybrids than in their parental lines. Volatile heterosis was obvious in both F1 hybrids. At the molecular level, the comparative transcriptomics analysis revealed that approximately 41% (9027 of 21,995) of the genes showed non-additive expression, whereas only 7.83% (1723 of 21,995) showed additive expression. Among the non-additive genes, 42.1% showed high parental dominance and 17.6% showed over-dominance. Among different expression genes with high parental dominance and over-dominance expression patterns, KEGG and GO analyses found that plant hormone signal transduction, tea plant physiological process related pathways and most pathways associated with tea tree volatiles were enriched. In addition, we identified multiple genes (CsDXS, CsAATC2, CsSPLA2, etc.) and transcription factors (CsMYB1, CsbHLH79, CsWRKY40, etc.) that played important roles in tea volatile heterosis. Based on transcriptome and metabolite profiling, we conclude that non-additive action plays a major role in tea volatile heterosis. Genes and transcription factors involved in tea volatiles showing over-dominance expression patterns can be considered candidate genes and provide novel clues for breeding high-volatile tea varieties.

In this study, complementary metabolomic and proteomic analyses were conducted on the solar- and indoor-withered oolong tea leaves, and freshly plucked leaves as the control, for the purpose to reveal the mechanisms underlying the initial formation of some flavor determinants during the early stage of oolong tea processing. As a result, a total of 978 non-volatile compounds and 152 volatile compounds were identified, the flavonoids and several esters were differently accumulated in various tea samples. In total, 7048 proteins were qualitatively and quantitatively determined, the analysis on pathway enrichment showed that phenylpropanoid, flavonoid metabolisms, and protein processing in endoplasmic reticulum were the major pathways discriminating the different tea samples. The joint protein–metabolite analysis showed that the multiple stresses such as dehydration, heat, and ultra-violet irradiation occurred during the withering step induced the dynamic and distinct changes in the biochemical network in the treated leaves compared to fresh leaves. The significant decreases in flavonoids, xanthine alkaloids, and several amino acids contributed to the alleviation of bitter or astringent taste of withered leaves, although the decomposition of L-theanine resulted in the loss of umami flavor over the solar-withering step. Moreover, the fruity or floral aromas, especially volatile terpenoids and phenylpropanoids/benzenoids, were retained or accumulated in the solar withered leaves, potentially aiding the formation of a better characteristic flavor of oolong tea made by indoor withered tea leaves. Distinct effects of solar- and indoor-withering methods on the flavor determinant formation provide a novel insight into the relationship between the metabolite accumulation and flavor formation during the withering step of oolong tea production.