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CamK-DB: A k-mer MinHash fingerprint database for reference-free genotyping of Camellia accessions.

Tea (Camellia sinensis L.), a major global economic crop in Asia, poses challenges for genetic identification because its highly heterozygous, repetitive genome reduces the efficacy of conventional single-nucleotide polymorphism (SNP) and microsatellite markers, and interspecific hybridization further complicates the situation. To address these issues, CamK-DB was developed as a reference-free Camellia fingerprinting database built on MIKE MinHash sketches. We curated 418 candidate resequencing datasets, and built a database using standardized 5× genome-coverage fingerprints. Each accession is stored as a MIKE. jac fingerprint generated with k = 21 and recommended sketch/pre_cnt = 2000. CamK-DB provides a command-line interface for data management and a custom C++ query engine that computes top-10 matches using Jaccard similarity, complemented by a QT-based graphical interface for interactive analysis. This resource offers a robust and scalable framework for precise and routine germplasm identification, genomic phylogenetic inference, and strategic breeding program design. CamK-DB (database and code) is publicly available at https://github.com/sc-zhang/CamK-DB. CamK-DB binaries are provided for Windows 10/11 and Linux (x86_64, glibc ≥ 2.27).

Databases, Genetic

Streptomyces xingningensis sp. nov. and Streptomyces rhizosphaerae sp. nov. isolated from the rhizosphere soil of Camellia oleifera.

Two novel actinobacteria strains, designated YCB024T and YCB041T, were isolated from rhizosphere soil samples of Camellia oleifera collected from Xingning City, Guangdong Province, PR China. Both strains exhibited mycelial filaments and intertwined hyphal cells. Phylogenetic analysis revealed that strain YCB024T exhibited the highest 98.8% 16S rRNA gene similarity with Streptomyces yogyakartensis NBRC 100779T, Streptomyces javensis NBRC 100777T and Streptomyces violaceusniger NBRC 13459T, and strain YCB041T exhibited the highest 98.9% similarity with Streptomyces albospinus NBRC 13846T. The average nucleotide identities (ANI) and digital DNA-DNA hybridization (dDDH) values among the two strains and their closely related species indicated that they were clearly different from other known Streptomyces species. The draft genome sizes of the two strains were 6.9 Mbp and 9.5 Mbp with DNA G+C contents of 70.4 mol% and 71.7 mol%. The major cellular fatty acids in strain YCB024T included iso-C14 : 0, iso-C15 : 0, anteiso-C15 : 0, iso-C16 : 0 and C16 : 0, and strain YCB041T included iso-C15 : 0, anteiso-C15 : 0, iso-C16 : 0 and C16 : 0. The major polar lipids of the two strains were diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylinositol mannoside or unidentified aminophospholipid. The respiratory quinones of both strains included MK-9 (H6), MK-9 (H8) and MK-10 (H4). Based on phylogenetic analysis, ANI and dDDH values, physiological and chemical properties, strains YCB024T and YCB041T represent two novel species of the genus Streptomyces, for which the names Streptomyces xingningensis sp. nov. (type strain YCB024T=GDMCC 4.314T=JCM 36249T) and Streptomyces rhizosphaerae sp. nov. (type strain YCB041T=GDMCC 4.469T=JCM 38170T) are proposed, respectively.

Streptomyces

Systematic Identification and Functional Characterisation of Colletotrichum fructicola Effectors During Camellia oleifera Colonisation.

Camellia oleifera is an important woody oil crop in southern China, but its production is severely threatened by anthracnose caused by Colletotrichum fructicola. C. fructicola deploys secreted effector proteins to establish infection. However, systematic identification and functional characterisation of C. fructicola effector genes upregulated during infection remains largely unexplored. Here, we integrated genome-wide secretome prediction with RNA-seq data from C. oleifera leaves inoculated with C. fructicola to identify candidate effectors induced during infection, followed by functional screening, targeted gene deletion, complementation and pathogenicity assays. Five novel effectors required for C. fructicola full virulence were identified, all of which suppressed Bax-induced cell death in Nicotiana benthamiana. Targeted deletion of the corresponding genes in C. fructicola reduced lesion areas by 47%-78% on C. oleifera leaves and by up to 67% on apple fruits, whereas complementation restored their virulence to wild-type levels. Transcriptomic profiling of infected postharvest C. oleifera fruits identified differentially expressed genes enriched in GO terms related to copper ion response, as well as in KEGG pathways associated with phenylpropanoid biosynthesis, taurine and hypotaurine metabolism, and plant-pathogen interactions. In addition, superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) activities, as well as malondialdehyde content, were altered in C. oleifera leaves inoculated with the effector-deletion mutants compared with those infected with the wild-type strain, suggesting that these effectors may contribute to the modulation of host oxidative stress responses. Taken together, our findings provide genetic and physiological evidence that these effectors contribute to C. fructicola virulence, offering potential targets for anthracnose control.

Colletotrichum

Genome-wide identification and evolutionary analysis of the ERF-VII gene family in the tea plant (Camellia sinensis) and functional characterization of CsRAP2.2 in response to cold stress.

The ERF-VII gene family, a critical branch of the AP2/ERF superfamily, is central to plant stress adaptation. However, its evolutionary history and function in tea plant (Camellia sinensis) remain unclear. Here, we performed integrated evolutionary, genomic, and functional analyses of ERF-VII genes across 14 plant lineages and 20 tea plant cultivars. The phylogenetic analysis revealed that ERF-VII proteins originated after vascular plant divergence, coinciding with the emergence of the N-terminal MCGGA/I motif linked to the oxygen-dependent N-degron pathway. Gymnosperms retained few conserved members, whereas angiosperms exhibited lineage-specific expansion-extensive in monocots via whole-genome duplication, moderate in eudicots with functional diversification. Pan-genome analysis across 20 tea plant cultivars further revealed varietal differences in ERF-VII gene distribution. Transcriptome profiling via the Tea Plant Information Archive identified CsRAP2.2 as a cold-inducible ERF-VII member with sustained expression under low-temperature stress. Functional assays demonstrated that silencing CsRAP2.2 reduced cold tolerance, while overexpression in tea leaves and heterologous expression in Arabidopsis thaliana enhanced cold tolerance by maintaining photosystem II efficiency, reducing membrane lipid peroxidation, and improving antioxidant capacity. Weighted gene co-expression network analysis positioned CsRAP2.2 as a regulatory hub integrating cold, hormone, and oxygen-sensing pathways. These results clarify the evolutionary trajectory of ERF-VII genes and establish CsRAP2.2 as a core cold-tolerance regulator in tea plant. These findings may inform future breeding of cold-resilient tea cultivars.

Camellia sinensis

Establishment of a cBSA-mediated miRNA delivery system in Camellia sinensis and functional validation of the Cs-miR163/CsSK1 module in cold stress response.

Cold stress severely limits tea (Camellia sinensis) yield and quality. MicroRNAs (miRNAs) are key post-transcriptional regulators of plant cold responses; however, in vivo functional validation in tea plants is hindered by the lack of efficient genetic transformation and nucleic acid delivery systems. In this study, a cationized bovine serum albumin (cBSA)-mediated miRNA delivery system was established in tea plants. The cold-responsive miRNA Cs-miR163 and its target gene CsSK1 (a negative regulator of cold tolerance) were used as a model. Direct cleavage of CsSK1 mRNA by Cs-miR163 was confirmed by 5' RLM-RACE and GUS transient expression assays, and enhanced cold tolerance was demonstrated in Arabidopsis overexpression lines. The cBSA preparation protocol was optimized, yielding stable cBSA/miRNA complexes with high protective capacity across temperatures of 15-35 °C and pH 4.5-7.2. Delivery parameters were systematically evaluated; optimal conditions were determined as 2 mg/mL cBSA with 10 nM miRNA and solution uptake into 3-cm cuttings for 5 days, enhancing miRNA delivery efficiency by approximately 48-fold. Transmission electron microscopy provided direct ultrastructural evidence that cBSA/miRNA nanocomplexes are internalized into tea plant cells via adsorptive-mediated endocytosis involving electrostatic membrane adsorption, membrane invagination, and cytoplasmic release. Under optimized conditions, cBSA-mediated delivery of Cs-miR163 silenced CsSK1 expression by approximately 72%, reduced relative electrolyte leakage and ROS accumulation, and markedly enhanced cold tolerance. The regulatory role of the Cs-miR163/CsSK1 module was clarified, and the established system provides a promising strategy for functional genomics in woody plants that warrants further testing in additional species and tissues.

Camellia sinensis

Mining of CtOPR2 as an essential gene regulating jasmonic acid mediated insect tolerance in Camellia tachangensis through genome-wide association studies.

Jasmonic acid (JA), a pivotal lipid-derived phytohormone, serves as a critical regulator in plant growth and defense mechanisms. However, the genetic mechanisms of OPR2 gene in JA-dependent biotic defenses of Camellia tachangensis have rarely been investigated. In this study, we performed a genome-wide association study to analyze 100,720 high-quality single nucleotide polymorphisms (SNPs) among 350 tea accessions from Guizhou province to identify genetic variations associated with JA. Analysis showed C. tachangensis displayed higher levels of JA content, further analysis identified 60 high-quality SNPs and nine candidate genes related to JA. Among them, CtOPR2 encoding 12-oxophytodienoic acid reductase 2 is responsible for catalyzing the conversion of 4,5-didehydro-JA (4,5-ddh-JA) to JA. The expression level of CtOPR2 in three tea accessions with different JA content was consistent with the dynamic changes of JA content. The expression level of synthetic (AOS, AOC, and ACX) and responsive (WRKY18 and MYC2) genes were significantly decreased and increased in asODN-CtOPR2-treated shoot tips and transgenic tobacco lines overexpressing CtOPR2, respectively, which were consistent with the JA content. These results further revealed that CtOPR2 gene played essential roles in promoting JA biosynthesis. A significant reduction in insect bite area was observed on transgenic tobacco leaves compared to wild-type leaves in feeding experiments with Spodoptera litura, highlighting that the positive regulatory function of CtOPR2 gene in JA-mediated immune responses. This study provides a robust theoretical foundation for marker-assisted selection breeding in tea, aimed at developing high-JA germplasm with potentially enhanced pest resistance for cultivation in Guizhou.

Journal Article

Carcinogenicity of Camellia sinensis (tea) and some tannin-containing folk medicinal herbs administered subcutaneously in rats.

In an attempt to correlate the high incidence of esophageal carcinoma in natives of certain places with their habit of using herbaceous folk medicines, we performed bioassays of several plant extracts and the fractions prepared from them. Fourteen extracts and fractions from 6 plants were injected sc into NIH Black rats. The tannin fractions from Quercus falcata pagodaefolia, Diospyros virginiana, and Camellia sinensis were very active and produced tumors at the injection site in 66% or more of the treated animals. Tannin fractions from 3 other plants and total aqueous extracts from 5 to 6 tested plants were also tumorigenic rats. The induced tumors were malignant fibrous histiocytomas similar, if not identical, to those encountered in humans. The experiment indicated a possibility of induction of tumor in man by the tested plant materials.

Animals

CsMYB219 and CsMYB196 influence epigallocatechin gallate biosynthesis in tea plant (Camellia sinensis) by regulating CsSCPL1A gene expression.

Epigallocatechin gallate (EGCG) is the most abundant and biologically active catechin in tea leaves and has been widely utilized in the development of functional foods. EGCG is catalyzed by serine carboxypeptidase-like 1A (CsSCPL1A) acyltransferases in tea plants. Although CsSCPL family genes are regulated by several transcription factors (TFs), systematic studies on their regulation by MYB TFs are lacking. This study integrates targeted metabolomics, transcriptomics, DNA-protein, and protein-protein interaction analyses to elucidate the transcriptional regulation of EGCG biosynthesis-related genes CsSCPL4 and CsSCPL5-1 by R2R3-MYB TFs. CsMYB219 and CsMYB196 can specifically bind to CsSCPL4 and CsSCPL5-1 promoters and activate their expression. CsMYB196 also interacted with CsTT8a and CsTTG1 to activate the transcription activity of CsSCPL4 and CsSCPL5-1 promoters by forming a MYB/bHLH/WD40 (MBW) complex. Promoter truncation assays delineated MYB-responsive cis-elements in CsSCPL4 (-613 to -1 bp with enhancers at -1967 to -1622) and CsSCPL5-1 (-503 to -296 bp). Silencing of CsMYB219 and CsMYB196 by virus-induced gene silencing (VIGS) assay significantly reduced the expression levels of CsSCPL4 and CsSCPL5-1 and EGCG content in tea leaves. Transient overexpression of CsMYB219 and CsMYB196 in tea leaves upregulated CsSCPL4 and CsSCPL5-1 expression and elevated EGCG content. These findings enhance our understanding of the regulatory network underlying EGCG biosynthesis in tea plants and provide a solid foundation for future genetic improvement of tea plant cultivars.

Catechin

Systematic mining and characterization of metal transporter families regulating zinc homeostasis provide insights into metal homeostasis in Camellia sinensis.

BACKGROUND AND AIMS: Zinc is essential for tea plant growth and quality formation, yet its homeostatic mechanisms remain poorly understood. This study identified metal transporter families regulating zinc homeostasis, analyzed their evolution, structure, and expression, and clarified zinc uptake, transport, detoxification networks, and their links to metabolism. METHODS: This study identified zinc homeostasis-related metal transporter families in the tea plant genome, characterized their structural features and expression profiles across tissues and developmental stages through integrative bioinformatics and transcriptomic analyses, and delineated the molecular mechanisms underlying zinc uptake, translocation, and detoxification by systematically integrating published evidence. RESULTS: This study identified 74 metal transporter genes from six families: 13 CsZIPs, 12 CsNRAMPs, 10 CsHMAs, 10 CsYSLs, 14 CsMTPs, and 15 CsCAXs in the 'Shuchazao2' genome, revealing closer affinity to woody species than to Arabidopsis. These proteins exhibit conserved domains, diverse subcellular localizations (cell membrane, vacuole, chloroplast, and Golgi apparatus), and tissue-specific expression with abundant stress/hormone-responsive cis-elements. At the plant-soil interface, tea plants mobilize rhizospheric zinc via proton and organic acid secretion; CsYSLs, CsNRAMPs, and CsZIPs mediate zinc uptake, aided by arbuscular mycorrhizal fungi (AMF) and plant growth-promoting rhizobacteria (PGPR) that expand root absorption zones. Xylem CsHMAs and phloem CsYSLs coordinate root-to-shoot zinc translocation, and vacuolar transporters (CsMTPs, CsCAXs), cell wall immobilization, and antioxidant systems alleviate high-zinc stress injury. CONCLUSIONS: These findings collectively delineate an integrated zinc "acquisition-distribution-buffering" network in tea plants, offering a repertoire of candidate genes with potential utility in zinc biofortification breeding and improving acid soil adaptation. Further experimental validation, including tea transgenesis, zinc-stress qRT-PCR, and heterologous functional complementation, is essential to substantiate their biological roles.

Camellia sinensis

Further investigation of the biosynthesis of caffeine in tea plants (Camellia sinensis L.). Methylation of transfer ribonucleic acid by tea leaf extracts.

1. The tRNA methyltransferase activity in vitro of leaves, cotyledons and roots of 85-day-old tea seedlings was studied. 2. The activity of extracts prepared from tea leaves with Polycar AT (insoluble polyvinylpyrrolidine) had optimum pH7.7 and was greatly influenced by thiol compounds, but only slightly by metal ions and ammonium acetate. 3. The activities of extracts, expressed per mg of protein, were as follows: roots greater than leaves greater than cotyledons. The only methylated base isolated after incubation with these preparations was 1-methyladenine. 4. The results did not support the view of involvement of methylation of nucleic acids in caffeine biosynthesis in tea plants. In contrast, it is suggested that theophylline is synthesized from the specific methylated precursor in nucleic acids, namely 1-methyladenylic acid, via 1-methylxanthine.

Caffeine

Dietary fat-dependent changes in hepatic cholesterogenesis and the activity of 3-hydroxy-3-methylglutaryl-CoA reductase in fasted-refed rats.

Effects of various dietary fats on the activity of 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase and sterol and fatty acid synthesis from [1-14C]acetate and [2-14C]mevalonate were examined in the liver from fasted-refed rats. Rats fasted for 2 days were refed a fat-free diet or diets containing various fats (tricaprylin, trilaurin, trimyristin, tristearin, camellia oil, or safflower oil) at the 10% level for 1, 3, or 7 days. The activity of HMG-CoA reductase was restored to about one-half of the pre-fasting levels in all groups after refeeding for 1 day and increased to above the pre-fasting levels after 3 days, with the exception of safflower oil, the rise was especially noticeable when fat-free, tricaprylin, and tristearin diets were fed. After 7 days, the activity of HMG-CoA reductase, except for rats refed tristearin, was decreased to levels that were far below those observed after 1 day-refeeding. This was particularly marked with tricaprylin, trilaurin, and camellia oil. The response of sterogenesis resembled that of the reductase. Dietary fat-dependent modification of fatty acid synthesis from [1-14C]acetate was first demonstrated after 7 days. Hepatic esterified cholesterol tended to accumulate and the deposition was marked after 3 days of refeeding. However, fat-dependent alterations of this parameter were remarkable on day 7. The concentration of plasma cholesterol also showed dietary fat-dependent changes after refeeding. Dietary fats appear to play an important role not only in the regulation of hepatic HMG-CoA reductase and sterol synthesis, but also in the overall processes of cholesterol dynamics.

Animals

Alginate-based edible coating incorporating green tea extract for preserving postharvest quality and safety of white mushrooms (Agaricus bisporus).

This study aimed to evaluate the effects of a sodium alginate based edible coating incorporated with green tea extract (GTE) (Camellia sinensis) on the postharvest quality attributes and antimicrobial activity against Listeria monocytogenes in white mushrooms during refrigerated storage. The phenolic profile of GTE was characterized, and its minimum inhibitory concentration (MIC) against L. monocytogenes (1.6&#xa0;mg/mL) was determined. Sodium alginate coatings, with (ALG-GTE) or without GTE (ALG) at MIC (1.6&#xa0;mg/mL), were characterized (functional groups, solubility in water, moisture, thickness, water contact angle and color) for their chemical and physical properties. The effects of ALG-GTE coatings on quality parameters (firmness, weight loss, color, pH, sugars and organic acids), enzymatic activity [polyphenol oxidase (PPO), peroxidase (POD) and pectin methylesterase (PME)], antimicrobial activity against L. monocytogenes (5 log CFU/g), and surface characteristics (3D optical profilometry) were assessed in white mushrooms (Agaricus bisporus) during refrigerated storage (8&#xa0;days, 4&#xa0;&#xb1;&#xa0;1&#xa0;&#xb0;C, 90-95% RH). The ALG-GTE coatings preserved sugar composition, particularly rhamnose, reduced organic acids accumulation and delayed weight and firmness loss, reduced color changes, and decreased PME activity in coated white mushrooms. L. monocytogenes counts decreased by 1.4 log CFU/g after 1&#xa0;day, and no viable cells were detected after 2&#xa0;days (< 1.5 log CFU/g) in ALG-GTE coated white mushrooms. In addition, ALG-GTE coated white mushrooms exhibited smoother surfaces than uncoated samples. These findings highlight the potential of ALG-GTE coatings as a sustainable alternative capable of improving the microbiological safety and delaying the postharvest changes in fresh mushrooms.

Agaricus

Regulation by dietary fats of 3-hydroxy-3-methylglutaryl-Coenzyme A reductase in rat liver.

The effects of various dietary fats on the activity of 3-hydroxy-3-methylglutaryl-Coenzyme A (HMG-CoA) reductase in rat liver microsomes, the rate-limiting enzyme in cholesterogenesis, were examined. A series of experiments demonstrated the dependency of the HMG-CoA reductase activity on the nature of dietary fats. When saturated fats with chain length of 12 to 18 were the dietary sources and were fed at the 10% level for 19 days, feeding fats with shorter chain fatty acids caused a lower enzyme activity compared to those with longer chain fatty acids. The activity was also regulated by the degree of unsaturation of dietary fats; when safflower oil, camellia oil or tristearin were fed at the 10% level for 18 days, the higher the unsaturation, the lower the activity. When trimyristin or tripalmitin were fed at the 10% level for 14 days, addition of essential fatty acid, at the level of minimum daily requirement (1% was replaced by safflower oil), did not affect the enzyme activity. Through the rate of incorporation of mevalonate into cholesterol in the 12,500 x g supernatant fraction of the liver was also found to be influenced by the types of dietary fats, the extent of the response appeared much smaller than that of HMG-CoA reductase. No consistent correlation between the HMG-CoA reductase activity and the content of microsomal cholesterol or cholesteryl ester and the fatty acid composition of microsomal lipids was observed.

Animals

[The effect of rooibos tea on iron absorption].

A study was carried out to determine if rooibos tea (Aspalathus linearis) has a deleterious effect on iron absorption similar to that of ordinary tea (Camellia sinensis). Three groups of volunteers, each consisting of 10 healthy young men comparable with regard to iron status and body dimensions, were studied. After ingestion of 1 muCi 59Fe and 16 mg of elemental iron, group A drank rooibos tea, group B ordinary tea and group C boiled water. Iron absorption was measured 14 days later with a whole body counter. Mean iron absorption after ingestion of rooibos tea, ordinary tea, and water was 7,25%, 1,70% and 9,34% respectively. In contrast to ordinary tea (P less than 0,0001), rooibos tea did not affect iron absorption significantly. The ascorbic acid content of rooibos tea varies between 121,8 and 154,9 mumol/l, depending on the method of preparation. Rooibos tea contains small amounts of iron, magnesium, phosphorus, sodium, chloride and potassium. These findings have practical nutritional and therapeutic implications.

Adult

Integrated metabolomic, transcriptomic, and proteomic analyses reveal changes in the non-volatile metabolite profile of LED light-withered oolong tea.

LED light withering is a crucial method for overcoming weather limitations and enhancing the quality of oolong tea. To elucidate the underlying molecular mechanisms, this study simulated solar spectra using multiwavelength LED light and compared the resulting metabolic, transcriptomic, and proteomic profiles during the enzymatic-catalysis process (ECP) in oolong tea processing. Results indicated that LED light withering altered gene expression and protein regulation of secondary metabolism, particularly in the flavonoid biosynthesis pathway. These shifts encompassed key quality-related compounds, including flavonoids (quercetin-3-O-rhamnoside, dihydroquercetin), amino acids (L-asparagine, L-histidine), guanosine 5'-monophosphate (GMP), and carbohydrates. Furthermore, LED light withering accelerated tea leaf water loss, influenced gene expression involved in photosynthetic cellular components (chloroplasts, thylakoids), increased ascorbate peroxidase regulation under stress, and subsequently modulated energy metabolism and signal transduction in tea leaves. This study offers molecular theoretical framework for the controlled light-withering of oolong tea under bad weather and the associated improvements in its quality.

Camellia sinensis

Multi-omics insights into aroma formation in congou black tea during fermentation.

This study used multi-omics technologies to analyze aroma formation during Congou black tea fermentation. Volatile compounds were analyzed by headspace solid phase microextraction coupled with gas chromatography mass spectrometry using two columns of different polarity. Fermentation increased total volatile normalized peak area fivefold, with alcohols, aldehydes, and acids increasing over sevenfold. 29 differential metabolites were screened, including amino acid derived phenylacetaldehyde, phenylethanol, and 2-methylbutanal; fatty acid derived (E,E)-2,4-heptadienal, hexanal, and 1-hexanol; and isoprenoid derived linalool, geraniol, and beta ionone. Transcriptomic, proteomic, and enzyme analyses revealed that biosynthesis contributed to early accumulation of amino acid and isoprenoid derived aromas, whereas ortho quinone mediated Strecker degradation and free radical induced fatty acid auto oxidation dominated generation of amino and fatty acid derived aromas during middle and late fermentation. In conclusion, aroma formation during fermentation results from biosynthesis and non-enzymatic oxidation, with the latter possibly dominating amino and fatty acid derived aromas.

Fermentation

Enhancement flavor quality in Zhao'an Baxian oolong tea through enhanced turning-over process.

A systematical investigation on the effects of turning-over intensity on the flavor formation of Zhao'an Baxian oolong tea (ZBT) was performed, through a comparative analysis of heavy turning-over (HT) and light turning-over (LT) treatments in this study. The tea samples were subjected to proteomic and metabolomic analyses, combined with quantitative descriptive analysis (QDA) and electronic sensory (E-tongue/E-nose) evaluation. The results demonstrate that HT significantly reduced the content of bitter and astringent compounds, such as catechins and flavonol glycosides, while promoting the accumulation of umami-related amino acids. Concurrently, HT enhanced the biosynthesis of key floral and fruity volatiles, such as &#x3b2;-ocimene, geraniol, benzaldehyde, jasmone by activating stress-responsive metabolic pathways. These coordinated biochemical changes, driven by enzyme-catalyzed reactions in response to prolonged mechanical wounding and environmental stress, collectively improved the overall sensory profile of ZBT. These findings provide a mechanistic foundation for improving ZBT production, with clear implications for quality control and flavor-oriented product development.

Tea

Cis-regulatory evolution of CsANS1 drives cultivar variation in anthocyanin accumulation in tea plants.

Anthocyanins, a ubiquitous class of water-soluble phytochemicals renowned for their chromatic diversity and potent bioactivity, are integral to the phenotypic and metabolic plasticity of higher plants. Using an integrative multi-omics approach that combines transcriptomic and metabolomic profiling, we identified anthocyanin synthase (CsANS1) as the key genetic determinant responsible for interspecific variation in anthocyanin accumulation among tea plants. Architectural comparison of promoter regions revealed a 192-bp variation insertion in the CsANS1 cis-regulatory region with potential functional significance. This insertion was strictly conserved in anthocyanin-rich (purple-leaf) cultivars, including both natural and hybrid genotypes, but entirely missing in anthocyanin-deficient (green-leaf) cultivars. Dual-luciferase assays confirmed that this insertion enhances promoter activity. Additionally, we delineated a tripartite regulatory axis comprising CsmiR156b, CsSPL9, and CsMYB75 which orchestrates the spatiotemporal modulation of CsANS1 expression and, consequently, anthocyanin biosynthesis. Collectively, these findings provide a mechanistic paradigm for anthocyanin polymorphism in tea plants, implicating both cis-regulatory evolution and transcriptional network synergy as pivotal drivers of phytochemical diversification.

Anthocyanins