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As important factors to oolong tea quality, the accumulation and dynamic change in aroma substances attracts great attention. The volatile composition of oolong tea is closely related to the precursor contents. Fatty acids (FAs) and their derivatives are basic components of oolong tea fragrance during the postharvest process. However, information about the precursors of FAs during the postharvest process of oolong tea production is rare. To investigate the transformation of fatty acids during the process of oolong tea production, gas chromatograph–flame ionization detection (GC-FID) was conducted to analyze the composition of FAs. The results show that the content of total polyunsaturated FAs initially increased and then decreased. Specifically, the contents of α-linolenic acid, linoleic acid and other representative substances decreased after the turn-over process of oolong tea production. The results of partial least squares discrimination analysis (PLS-DA) showed that five types of FAs were obviously impacted by the processing methods of oolong tea (VIP > 1.0). LOX (Lipoxygenase, EC 1.13.11.12) is considered one of the key rate-limiting enzymes of long-chain unsaturated FAs in the LOX-HPL (hydroperoxide lyase) pathway, and the mechanical wounding occurring during the postharvest process of oolong tea production greatly elevated the activity of LOX.

Aroma is an important index of tea quality. The volatile C6‐compounds formed from linoleic and linolenic acids in tea leaf lipids are essential components of tea. C6‐compounds are formed and transformed during the postharvest process of tea leaves. However, the metabolic flux of these C6‐compounds, the activities of related enzymes, and the transcription of related genes during the postharvest process of oolong tea remain unclear. In this study, the chemical profiles of C6‐aldehydes and C6‐alcohols, the pattern of ADH enzyme activity, and the level of CsADH gene expression during the postharvest process of oolong tea were investigated. We found that the turnover process had a positive effect on the accumulation of C6‐alcohols and simultaneously induced ADH activity, especially during the withering stage. The expression of CsADH peaked during the turnover stage. The relative expression level of CSA019598 typically increased during the postharvest process. Correlation analysis demonstrated that CSA019598 expression increased as ADH activity increased. This finding suggests that CSA019598 may play a prominent role in regulating ADH. These results advance our understanding of C6‐compound formation during the postharvest process of oolong tea. We aim to evaluate how green leaf volatiles affect the enzymatic formation and genetic transcription of aromatic compounds in oolong tea in future studies., (a) ADH reduces C6‐Aldehydes into C6‐alcohols during the postharvest process of oolong tea under mechanical wounding conditions. (b) CSA019598 screened from tea plant genome might be an available CsADH gene which plays a major regulatory role during the postharvest process of oolong tea.

Aroma is an essential quality indicator of oolong tea, a tea derived from the Camellia sinensis L. plant. Carboxylic 6 (C6) acids and their derivative esters are important components of fatty acid (FA)-derived volatiles in oolong tea. However, the formation and regulation mechanism of C6 acid during postharvest processing of oolong tea remains unclear. To gain better insight into the molecular and biochemical mechanisms of C6 compounds in oolong tea, a combined analysis of alcohol dehydrogenase (ADH) activity, CsADH2 key gene expression, and the FA-derived metabolome during postharvest processing of oolong tea was performed for the first time, complemented by CsHIP (hypoxia-induced protein conserved region) gene expression analysis. Volatile fatty acid derivative (VFAD)-targeted metabolomics analysis using headspace solid-phase microextraction–gas chromatography time-of-flight mass spectrometry (HS-SPEM-GC-TOF-MS) showed that the (Z)-3-hexen-1-ol content increased after each turnover, while the hexanoic acid content showed the opposite trend. The results further showed that both the ADH activity and CsADH gene expression level in oxygen-deficit-turnover tea leaves (ODT) were higher than those of oxygen-turnover tea leaves (OT). The C6-alcohol-derived ester content of OT was significantly higher than that of ODT, while C6-acid-derived ester content showed the opposite trend. Furthermore, the HIP gene family was screened and analyzed, showing that ODT treatment significantly promoted the upregulation of CsHIG4 and CsHIG6 gene expression. These results showed that the formation mechanism of oolong tea aroma quality is mediated by airflow in the lipoxygenase–hydroperoxide lyase (LOX-HPL) pathway, which provided a theoretical reference for future quality control in the postharvest processing of oolong tea.

Tea (Camellia sinensis (L.)) is an important economic plant. Light is the earliest external signal factor during the postharvest processing of oolong tea, and the solar withering is an indispensable process for aroma formation. In this study, Tieguanyin was used to analyze the effect of sunlight on aroma metabolism, which indicated that the main aroma compounds were significantly increased during solar withering for 15 min compared to the indoor withering. In addition, differentially expressed genes related to aroma metabolism were identified and quantified using the high-throughput Illumina RNA-Seq technology. The expression levels of key regulatory genes were consistent with the results from the gas chromatography-time of flight mass spectrometry (GC-TOF-MS) analysis, especially in terpenoid metabolic pathway, which showed that aroma metabolism could significantly respond to the short-term light, while its expression level was easily inhibited by the up-regulation of heat shock protein. Taken together, those data provides further insights into the mechanisms, contributing to aroma metabolism of tea plant.

Tea is one of three major non-alcoholic beverages that are popular all around the world. The economic value of tea product largely depends on the post-harvest physiology of tea leaves. The utilization of quantitative reverse transcription polymerase chain reaction is a widely accepted and precise approach to determine the target gene expression of tea plants, and the reliability of results hinges on the selection of suitable reference genes. A few reliable reference genes have been documented using various treatments and different tissues of tea plants, but none has been done on post-harvest leaves during the tea manufacturing process. The present study selected and analyzed 15 candidate reference genes: Cs18SrRNA, CsGADPH, CsACT, CsEF-1α, CsUbi, CsTUA, Cs26SrRNA, CsRuBP, CsCYP, CselF-4α, CsMON1, CsPCS1, CsSAND, CsPPA2, CsTBP. This study made an assessment on the expression stability under two kinds of post-harvest treatment, turn over and withering, using three algorithms—GeNorm, Normfinder, and Bestkeeper. The results indicated that the three commonly used reference genes, CsTUA, Cs18SrRNA, CsRuBP, together with Cs26SrRNA, were the most unstable genes in both the turn over and withering treatments. CsACT, CsEF-1α, CsPPA2, and CsTBP were the top four reference genes in the turn over treatment, while CsTBP, CsPCS1, CsPPA2, CselF-4α, and CsACT were the five best reference genes in the withering group. The expression level of lipoxygenase genes, which were involved in a number of diverse aspects of plant physiology, including wounding, was evaluated to validate the findings. To conclude, we found a basis for the selection of reference genes for accurate transcription normalization in post-harvest leaves of tea plants.