CsMYB73 negatively regulates theanine accumulation mediated by CsGGT2 and CsGGT4 in tea shoots (Camellia sinensis).

Theanine metabolism is a necessary biological process during the planting and production of tea that determines tea quality. There is currently little knowledge about the transcriptional regulation of theanine metabolism in tea plants. In this study, we demonstrated that γ-glutamyl-transpeptidase CsGGT4, as a homologous protein of the theanine hydrolase CsGGT2, exhibited a higher theanine synthesis catalytic efficiency. Homology modeling and molecular docking showed that differential protein structures between CsGGT2 and CsGGT4 implied their different biological functions in tea plants. Theanine content correlated significantly with the expression of CsGGT2, CsGGT4 and the transcription factor CsMYB73 in tea shoots from different seasons. Additionally, CsMYB73 was confirmed to act as a nucleus-localized transcription factor (TF), directly interacts with the CsGGT2 and CsGGT4 promoters, serving as an activator of CsGGT2 and a suppressor of CsGGT4. Consequently, this leads to a negative association with theanine accumulation in tea shoots. Furthermore, the continuous increase in CsMYB73 produced a significantly increase in CsGGT2 expression and inhibited CsGGT4 expression. The present study reveals that the degradation of theanine has been observed to increase, concomitantly with the inhibition of theanine synthesis, resulting in a significant decline in the accumulation of theanine in tea shoots during the process of seasonal greening in 'Huangkui' leaves. This study contributes to the broader comprehension of the intricate transcriptional regulatory hierarchy that governs the metabolism of theanine in tea shoots, offering novel approaches for managing tea plantations and enhancing tea quality.

Combined multi-omics approach to analyze the flavor characteristics and formation mechanism of gabaron green tea.

Gabaron green tea (GAGT) has unique flavor and health benefits through the special anaerobic treatment. However, how this composite processing affects the aroma formation of GAGT and the regulatory mechanism was rarely reported. This study used nontargeted metabolomics and molecular sensory science to overlay screen differential metabolites and key aroma contributors. The potential regulatory mechanism of anaerobic treatment on the aroma formation of GAGT was investigated by transcriptomics and correlation analyses. Five volatiles: benzeneacetaldehyde, nonanal, geraniol, linalool, and linalool oxide III, were screened as target metabolites. Through the transcriptional-level differential genes screening and analysis, some CsERF transcription factors in the ethylene signaling pathway were proposed might participate the response to the anaerobic treatment. They might regulate the expression of related genes in the metabolic pathway of the target metabolites thus affecting the GAGT flavor. The findings of this study provide novel information on the flavor and its formation of GAGT.

Role of Endophytic Bacteria in the Remobilization of Leaf Nitrogen Mediated by CsEGGT in Tea Plants (Camellia sinensis L.).

As an important economic plant, tea (Camellia sinensis) has a good economic value and significant health effects. Theanine is an important nitrogen reservoir, and its synthesis and degradation are considered important for nitrogen storage and remobilization in tea plants. Our previous research indicated that the endophyte CsE7 participates in the synthesis of theanine in tea plants. Here, the tracking test confirmed that CsE7 tended to be exposed to mild light and preferentially colonized mature tea leaves. CsE7 also participated in glutamine, theanine, and glutamic acid circulatory metabolism (Gln-Thea-Glu) and contributed to nitrogen remobilization, mediated by the γ-glutamyl-transpeptidase (CsEGGT) with hydrolase preference. The reisolation and inoculation of endophytes further verified their role in accelerating the remobilization of nitrogen, especially in the reuse of theanine and glutamine. This is the first report about the photoregulated endophytic colonization and the positive effect of endophytes on tea plants mediated and characterized by promoting leaf nitrogen remobilization.

A hierarchical model of ABA-mediated signal transduction in tea plant revealed by systematic genome mining analysis and interaction validation.

As a critical signaling molecule, ABA plays an important role in plant growth, development and stresses response. However, tea plant [Camellia sinensis (L.)], an important economical perennial woody plant, has not been systematically reported in response to ABA signal transduction in vivo. In this study, we mined and identified the gene structure of CsPYL/CsPP2C-A/CsSnRK gene families in the ABA signal transduction pathway through the genome-wide analysis of tea plants. Spatiotemporal expression and stress response (drought, salt, chilling) expression patterns were characterized. The results showed that most members of CsPYLs were conserved, and the gene structures of members of A-type CsPP2Cs were highly similar, whereas the gene structure of CsSnRK2s was highly variable. The transcription levels of different family members were differentially expressed with plant growth and development, and their response to stress signal patterns was highly correlated. The expression patterns of CsPYL/CsPP2C-A/CsSnRK2 gene family members in different tissues of tea plant cuttings after exogenous ABA treatment were detected by qRT-PCR, and the hierarchical model of ABA signaling was constructed by correlation analysis to preliminarily obtain three potential ABA-dependent signaling transduction pathways. Subsequently, the protein interaction of the CsPYL4/7-CsPP2C-A2-CsSnRK2.8 signaling pathway was verified by yeast two-hybrid and surface plasmon resonance experiments, indicating that there is specific selectivity in the ABA signaling pathway. Our results provided novel insights into the ABA-dependent signal transduction model in tea plant and information for future functional characterizations of stress tolerance genes in tea plant.