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Withering is one of the major processing steps critical for the quality of black tea. In this study, we investigated the mechanisms underlying the physicochemical changes in metabolites and gene expression during the withering process of black tea using metabolomic and transcriptomic approaches, respectively. Based on gas chromatography/mass spectrometry non-targeted metabolomic approaches (GC-MS) and ultra-high performance liquid chromatograph–tandem mass spectrometry (UHPLC-MS/MS), a total of 76 volatile compounds and 160 non-volatile compounds were identified from tea leaves, respectively. RNA-seq analysis revealed that the number of differentially expressed genes (DEGs) for the comparative combination of withering time (i.e., W4h, W6h, W8h, W10h, and W12h) compared with CK (i.e., fresh leaves) were 3634, 2906, 4127, 5736, and 7650, respectively. The core genes in starch metabolism, namely alpha-amylase (AMY) and beta-amylase (BAM), were upregulated as withering time increased. AMY and BAM contributed to the decomposition of starch to increase the soluble sugars. The content of tea leaf alcohols and aldehydes, which are the vital contributors for greenish aroma, gradually decreased as withering time increased due to the downregulation of associated genes while the compounds related to sweet and fruity characteristics increased due to the upregulated expression of related genes. Most DEGs involved in amino acids were significantly upregulated, leading to the increase in free amino acids content. However, DEGs involved in catechins metabolism were generally downregulated during withering, and resulted in a reduction in catechins content and the accumulation of theaflavins. The same trend was observed in alpha-linolenic acid metabolism-related genes that were downregulated and enhanced the reduction in grassy aroma in black tea. The weighted gene co-expression network analysis (WGCNA) of DEGs showed that one module can be associated with more components and one component can be regulated by various modules. Our findings provide new insights into the quality formation of black tea during the withering process.

This study delved into the aroma characteristics of “Qingxiang” oolong tea, analyzing six different cultivars and their processing modes. The findings showed that both cultivars and processing modes have a significant impact on the oolong tea aroma system. The study identified 18 terpenoid volatiles (VTs), 11 amino-acid-derived volatiles (AADVs), 15 fatty-acid-derived volatiles (FADVs), 3 carotenoid-derived volatiles (CDVs), and 10 other compounds in oolong tea that differentiate it from green and black tea. The turn-over stage was found to be the primary processing stage for oolong tea aroma formation. Molecular sensory analysis revealed that the “fresh” odor attribute is the basis for its aroma, while “floral and fruity” fragrances are its aroma characteristics. The perception of oolong tea as “fresh” and “floral and fruity” is influenced by the interactions of its aroma components. These findings provide a new basis for breed improvement and process enhancement in oolong tea production.

Nitrogen (N) is an important contributor in regulating plant growth and development as well as secondary metabolites synthesis, so as to promote the formation of tea quality and flavor. Theanine, polyphenols, and caffeine are important secondary metabolites in tea plant. In this study, the responses of Camellia sinensis roots to N deprivation and resupply were investigated by metabolome and RNA-seq analysis. N deficiency induced content increase for most amino acids (AAs) and reduction for the remaining AAs, polyphenols, and caffeine. After N recovery, the decreased AAs and polyphenols showed a varying degree of recovery in content, but caffeine did not. Meanwhile, theanine increased in content, but its related synthetic genes were down-regulated, probably due to coordination of the whole N starvation regulatory network. Flavonoids-related pathways were relatively active following N stress according to KEGG enrichment analysis. Gene co-expression analysis revealed TCS2, AMT1;1, TAT2, TS, and GOGAT as key genes, and TFs like MYB, bHLH, and NAC were also actively involved in N stress responses in C. sinensis roots. These findings facilitate the understanding of the molecular mechanism of N regulation in tea roots and provide genetic reference for improving N use efficiency in tea plant.

Abstract Background Heat stress factors (Hsfs) play vital roles in signal transduction pathways operating in responses to environmental stresses. However, Hsf gene family has not been thoroughly explored in tea plant (Camellia sinensis L.). Results In this study, we identified 25 CsHsf genes in C. sinensis that were separated by phylogenetic analysis into three sub-families (i.e., A, B, and C). Gene structures, conserved domains and motifs analyses indicated that the CsHsf members in each class were relatively conserved. Various cis-acting elements involved in plant growth regulation, hormone responses, stress responses, and light responses were located in the promoter regions of CsHsfs. Furthermore, degradome sequencing analysis revealed that 7 CsHsfs could be targeted by 9 miRNAs. The expression pattern of each CsHsf gene was significantly different in eight tissues. Many CsHsfs were differentially regulated by drought, salt, and heat stresses, as well as exogenous abscisic acid (ABA) and Ca2+. In addition, CsHsfA2 was located in the nucleus. Heterologous expression of CsHsfA2 improved thermotolerance in transgenic yeast, suggesting its potential role in the regulation of heat stress response. Conclusions A comprehensive genome-wide analysis of Hsf in C. sinensis present the global identification and functional prediction of CsHsfs. Most of them were implicated in a complex gene regulatory network controlling various abiotic stress responses and signal transduction pathways in tea plants. Additionally, heterologous expression of CsHsfA2 increased thermotolerance of transgenic yeast. These findings provide new insights into the functional divergence of CsHsfs and a basis for further research on CsHsfs functions.