Tea Plant Camellia Sinensis Research Articles

A New Colletotrichum Species Associated with Brown Blight Disease on Camellia sinensis.

Brown blight, as the most damaging and common foliar disease of the tea plant (Camellia sinensis) in China, has been recently reported to be caused by different species of the genus Colletotrichum. During the years 2016 to 2017, tea plants in commercial tea cultivation areas of Chongqing City that reported significant incidences of brown blight disease were investigated and then analyzed using both morphological characteristics and multilocus phylogenetic analysis. The results showed that at least five species of Colletotrichum were identified, including four well-known species (Colletotrichum gloeosporioides, C. camelliae, C. fioriniae, and C. karstii) and one novel species (C. chongqingense), indicating that there is remarkable species diversity in Colletotrichum spp. present as pathogens. Results of pathogenicity analyses confirmed that C. chongqingense was the causal agent of brown blight and different isolates differed in virulence. C. chongqingense, as a novel pathogen, has never been reported as being associated with brown blight disease in tea plants or anthracnose in other host plants anywhere in the world. Knowledge of the Colletotrichum populations will facilitate further studies addressing the relationships between Colletotrichum spp. and their host plant Camellia sinensis.

Metabolics and ionomics responses of tea leaves (Camellia sinensis (L.) O. Kuntze) to fluoride stress

Tea plant (Camellia sinensis (L.) O. Kuntze) is known to accumulate high concentrations of fluoride (F) in its leaves; however, the underlying mechanism of F accumulation remains unclear. The main objective of this study was to investigate the homeostatic self-defense mechanisms of tea leaves to F supplementation (0, 5, 20, and 50 mgL−1) by metabolomics and ionomics. We identified a total of 96 up-regulated and 40 down-regulated metabolites in tea leaves treated with F. Of these different compounds, minor polypeptides, carbohydrates and amino acids played valuable roles in the F-tolerating mechanism of tea plant. After F treatments, the concentrations of sodium (Na), ferrum (Fe), manganese (Mn), and molybdenum (Mo) were significantly increased in tea leaves, whereas the aluminum (Al) was decreased. These findings suggest that the ionic balance and metabolites are attributable to the development of F tolerance, providing new insight into tea plant adaptation to F stress.

Tea flavonoid theaflavin induces apoptosis in hepatocellular carcinoma cells through improving ROS level

Flavonoids, a class of normal polyphenolic mixes, restrain cell cycle movement and instigate apoptosis. This examination was performed to explore the anticancer effect of theaflavin, a natural flavonoid found in the leaves of tea plant Camellia sinensis. Although this molecule was found to inhibit several cancer cells, the specific anticancer action in liver cancer remains unexplored, especially in human hepatocellular carcinoma (HepG2) cells. Henceforth, the present study was designed to elucidate the anticancer activity in HepG2 cells, level of reactive oxygen species (ROS) in the cancer cells and tumour cell apoptosis. The action of theaflavin in provocation apoptosis was explored through the improved ROS by MTT assay and 2,7-dichlorodihydrofluorescein diacetate (DCFH-DA) method. As per the results obtained from the MTT assay, theaflavin had cell hindrance effect on HepG2 cells. The IC50 estimation of theaflavin to hindered cell development at 25, 50 and 75 μM concentration and instigating apoptosis through ROS improvement. The progressions in mitochondrial morphology, portion conditionally that diminished cell expansion, were seen in various concentrations of the drug treatment. In this manner, theaflavin might be useful as a chemotherapeutic agent for the treatment of liver cancer.

Roles of specialized metabolites in biological function and environmental adaptability of tea plant (Camellia sinensis) as a metabolite studying model

BackgroundTea is the second most popular beverage globally after water and contains abundant specialized metabolites. These metabolites give tea unique quality and are beneficial to human health. Some secondary metabolites are produced to help plants, including tea plants (Camellia sinensis), adapt to variable environment and grow normally. Therefore, whether abundant specialized metabolites have biological functions and play roles in the environmental adaptability of tea plants is of interest. Aim of reviewResearch progress regarding the biological functions of specialized metabolites (including catechins, l-theanine, caffeine, and volatile compounds) in tea plants is summarized. Furthermore, the main and characteristic scientific questions regarding tea plant growth in contrast to other economic crops are proposed, including (i) how tea plants adapt to acid soils, (ii) why tea plants have fewer diseases, and (iii) why tea plants and tea green leafhoppers have a symbiotic relationship. Accordingly, the potential adaptive mechanism is summarized, which is related to the function of specialized metabolites in tea plants. Key scientific concepts of reviewThis is the most in-depth investigation of biological functions of volatile compounds in tea plants. Direct in vivo evidence in tea plants shows that volatile compounds help defend against insects through plant-to-plant signaling. Furthermore, abundant specialized metabolites are speculated to contribute to the environmental adaptability of tea plants. However, further in vivo evidence and exploration of relevant mechanisms are required for all aspects discussed. This review provides an important reference for basic biological research on the tea plant as a specialized metabolite studying model.

Comparation study on the metabolism destination of neonicotinoid and organophosphate insecticides in tea plant (Camellia sinensis L.)

The absorption, distribution, metabolism and primary risk evaluation data of four neonicotinoids and two organophosphate insecticides in tea plant (Camellia sinensis L.) were compared. 22 neonicotinoid metabolites and 2 organophosphate metabolites were identified. The amount ratio of each neonicotinoid metabolite to its corresponding parent (M/P) was lower than 0.076 in the treated time. The organophosphates (omethoate and methamidophos) increased sharply, with M/Ps as high as 1.111 and 0.612. The risks evaluation of insecticides and their metabolites in treated leaves on day seven showed that the chronic risk was from the lowest 0.0759 (clothianidin) to highest 43.6409% (dimethoate), and the acute risk was highest 0.0370 for all targets. The calculated combined toxicity of leaves treated with acephate reached 1.5 folds in mature, 1.5 folds in tender leaves than no metabolites, and which of dimethoate were 2.1 folds in mature and 3.7 folds in tender leaves.

Genome-Wide Identification of the Tify Gene Family and Their Expression Profiles in Response to Biotic and Abiotic Stresses in Tea Plants (Camellia sinensis).

The TIFY family is a plant-specific gene family that is involved in regulating a variety of plant processes, including developmental and defense responses. The chromosome-level genome of the tea plant (Camellia sinensis) has recently been released, but a comprehensive view of the TIFY family in C. sinensis (the CsTIFY genes) is lacking. The current study performed an extensive genome-wide identification of CsTIFY genes. The phylogenetics, chromosome location, exon/intron structure, and conserved domains of these genes were analyzed to characterize the members of the CsTIFY family. The expression profiles of the CsTIFY genes in four organs were analyzed, and they showed different spatial expression patterns. All CsJAZ genes were observed to be induced by jasmonate acid (JA) and exhibited different responses to abiotic and biotic stresses. Six of seven CsJAZ genes (CsJAZ1, CsJAZ2, CsJAZ3, CsJAZ4, CsJAZ7, and CsJAZ8) were upregulated by mechanical wounding and infestation with the tea geometrid (Ectropis obliqua), while infection with tea anthracnose (Colletotrichum camelliae) primarily upregulated the expression levels of CsJAZ1 and CsJAZ10. In addition, CsJAZs were observed to interact with CsMYC2 and AtMYC2. Therefore, the results of this study may contribute to the functional characterization of the CsTIFY genes, especially the members of the JAZ subfamily, as regulators of the JA-mediated defense response in tea plant.

Metabolite signatures of diverse Camellia sinensis tea populations

The tea plant (Camellia sinensis) presents an excellent system to study evolution and diversification of the numerous classes, types and variable contents of specialized metabolites. Here, we investigate the relationship among C. sinensis phylogenetic groups and specialized metabolites using transcriptomic and metabolomic data on the fresh leaves collected from 136 representative tea accessions in China. We obtain 925,854 high-quality single-nucleotide polymorphisms (SNPs) enabling the refined grouping of the sampled tea accessions into five major clades. Untargeted metabolomic analyses detect 129 and 199 annotated metabolites that are differentially accumulated in different tea groups in positive and negative ionization modes, respectively. Each phylogenetic group contains signature metabolites. In particular, CSA tea accessions are featured with high accumulation of diverse classes of flavonoid compounds, such as flavanols, flavonol mono-/di-glycosides, proanthocyanidin dimers, and phenolic acids. Our results provide insights into the genetic and metabolite diversity and are useful for accelerated tea plant breeding.

Regulation of the Phenolic Compounds Accumulation in the Tea-Plant Callus Culture with a Separate and Combined Effect of Light and Cadmium Ions

The effect of light and cadmium ions (Cd) on the morphophysiological characteristics and accumulation of low molecular weight phenolic antioxidants in the tea plant Camellia sinensis L. callus culture under their separate and combined action is studied. It was established that the light exposure and the cadmium intake separately led to a change in the morphophysiological characteristics of the culture, a decrease in its growth, and activation of the antioxidant system. It was noted that it was manifested in an increase in the total content of phenolic compounds, flavans, and proanthocyanidins (to a lesser extent) and did not depend on the activity of the phenolic metabolism key enzyme, L-phenylalanine ammoniac lyase. It was found that the cell response was more pronounced under the action of Cd than under the action of light. In some cases, these indicators correlated with the level of lipid peroxidation in callus cultures. It is shown that all changes were more pronounced under the combined action of light and Cd.

Transcriptomic analysis of tea plant (Camellia sinensis) revealed the co-expression network of 4111 paralogous genes and biosynthesis of quality-related key metabolites under multiple stresses

The tea plant is an essential economic plant in many countries. However, its growing season renders them vulnerable to stresses. To understand the transcriptomic influences of these stresses on tea plants, we sequenced and analyzed the transcriptomes under drought, high-temperature, and pest. Paralogs were identified by comparing 14 evolutionarily close genomes. The differentially expressed paralog (DEPs) genes were analyzed regarding single or multiple stresses, and 1075 of the 4111 DEPs were commonly found in all the stresses. The co-expression network of the DEPs and TFs indicated that genes of catechin biosynthesis were associated with most transcription factors specific to each stress. The genes playing a significant role in the late response to drought and pest stress mainly functioned in the early response to high-temperature. This study revealed the relationship between stress and regulation of QRM synthesis and the role of QRMs in response to these (a)biotic stresses.

Transcription factor CsDOF regulates glutamine metabolism in tea plants (Camellia sinensis)

Glutamine plays a critical role in ammonium assimilation, and contributes substantially to the taste and nutritional quality of tea. To date, little research has been done on glutamine synthesis in tea plants. Here, a zinc finger protein CsDOF and a glutamine synthetase (GS)-encoding gene CsGS2 from tea plant (Camellia sinensis cv ‘Shuchazao’) were characterized, and their role in glutamine biosynthesis was determined using transient suppression assays in tea leaves and overexpression in Arabidopsis thaliana. The expression patterns of CsDOF and CsGS2, the GS activity and the glutamine content of photosynthetic tissues (leaf and bud) were significantly induced by shade. Suppressing the expression of CsDOF resulted in downregulated expression of CsGS2 and reduction of the leaf glutamine content. Moreover, in CsDOF-silenced plants, the expression of CsDOF and the glutamine content under shade treatment were higher than in natural light. The glutamine content and CsGS2 transcript level were also decreased in tea leaves when CsGS2 was suppressed, while they were higher under shade treatment than in natural light in CsGS2-silenced plants. In addition, the glutamine content and GS2 transcript level were increased when CsDOF and CsGS2 was overexpressed in Arabidopsis thaliana, respectively. In binding analyses, CsDOF directly bound to an AAAG motif in the promoter of CsGS2, and promotes its activity. The study shed new light on the molecular mechanism by which CsDOF activates CsGS2 gene expression and contributes to glutamine biosynthesis in tea plants.

A predicted protein functional network aids in novel gene mining for characteristic secondary metabolites in tea plant (Camellia sinensis)

Modeling a protein functional network in concerned species is an efficient approach for identifying novel genes in certain biological pathways. Tea plant (Camellia sinensis) is an important commercial crop abundant in numerous characteristic secondary metabolites (e.g., polyphenols, alkaloids, alkaloids) that confer tea quality and health benefits. Decoding novel genes responsible for tea characteristic components is an important basis for applied genetic improvement and metabolic engineering. Herein, a high-quality protein functional network for tea plant (TeaPoN) was predicted using cross-species protein functional associations transferring and integration combined with a stringent biological network criterion control. TeaPoN contained 31,273 nonredundant functional interactions among 6,634 tea proteins (or genes), with general network topological properties such as scale-free and small-world. We revealed the modular organization of genes related to the major three tea characteristic components (theanine, caffeine, catechin) in TeaPoN, which served as strong evidence for the utility of TeaPoN in novel gene mining. Importantly, several case studies regarding gene identification for tea characteristic components were presented. To aid in the use of TeaPoN, a concise web interface for data deposit and novel gene screening was developed (http://teapon.wchoda.com). We believe that TeaPoN will serve as a useful platform for functional genomics studies associated with characteristic secondary metabolites in tea plant.

Natural Variations and Dynamic Changes of Nitrogen Indices throughout Growing Seasons for Twenty Tea Plant (Camellia sinensis) Varieties.

Tea (Camellia sinensis (L.) O. Kuntze) leaves are harvested multiple times annually accompanied by a large amount of nitrogen (N) removed. Therefore, tea plantations are characterized by high requirements of N. This study aimed to assess the variations of N-level, apparent N remobilization efficiency (ANRE), and N utilization efficiency (NUtE) and their dynamic changes during growing seasons for twenty tea varieties. The N-level was highest in the one bud with two leaves as the youngest category, followed by mature leaves attached to green-red stems, and then by aging leaves attached to grey stems. The dynamic N-level presented different profiles of “S”-, “U”-, and “S-like”-shape in the three categories of leaves during the growing seasons. Here, specifically defined ANRE indicated N fluxes in a specific category of leaves, showing that sources and sinks alternate during the period of two consecutive rounds of growth. The dynamic of averaged NUtE followed an “S”-shape. The results revealed annual rhythms and physiological characters related with N indices, which were variety dependent and closely related with the amount of N requirements at proper time. An optimized NUtE is a complex character determined by the combination of tea plantation management and breeding practices to achieve sustainable development with economic benefit.

Genome-Wide Identification of CsATGs in Tea Plant and the Involvement of CsATG8e in Nitrogen Utilization.

Nitrogen (N) is a macroelement with an indispensable role in the growth and development of plants, and tea plant (Camellia sinensis) is an evergreen perennial woody species with young shoots for harvest. During senescence or upon N stress, autophagy has been shown to be induced in leaves, involving a variety of autophagy-related genes (ATGs), which have not been characterized in tea plant yet. In this study, a genome-wide survey in tea plant genome identified a total of 80 Camellia Sinensis autophagy-related genes, CsATGs. The expression of CsATG8s in the tea plant showed an obvious increase from S1 (stage 1) to S4 (stage 4), especially for CsATG8e. The expression levels of AtATGs (Arabidopsis thaliana) and genes involved in N transport and assimilation were greatly improved in CsATG8e-overexpressed Arabidopsis. Compared with wild type, the overexpression plants showed earlier bolting, an increase in amino N content, as well as a decrease in biomass and the levels of N, phosphorus and potassium. However, the N level was found significantly higher in APER (aerial part excluding rosette) in the overexpression plants relative to wild type. All these results demonstrated a convincing function of CsATG8e in N remobilization and plant development, indicating CsATG8e as a potential gene for modifying plant nutrient utilization.

Anti-skin aging activities of green tea (Camelliasinensis (L) Kuntze) in B16F10 melanoma cells and human skin fibroblasts

IntroductionSkin aging is a multifactorial process caused by chronological changes and environmental factors. Green tea, a processed tea plant (Camellia sinensis (L) Kuntze), possesses health benefits and has been used in traditional medicines and natural products. However, the anti-skin aging effect of green tea in clinical trials has been a controversial issue. To clarify this without the interference of systemic involvement, the objectives of study were to investigate the activities of green tea against skin aging in B16F10 melanoma cells and human skin fibroblasts. MethodsGreen tea leaves were extracted to standardise the phenolic profile. The cytotoxicity and the anti-skin aging activities, including melanogenesis assay, antioxidant activity, collagen content analysis, and matrix metalloproteinase-2 inhibitory assay, were evaluated in cell culture. ResultsGreen tea in this study composed mainly of epigallocatechin gallate (1,903.77 ± 33.59 μg/mL), epigallocatechin (930.24 ± 27.04 μg/mL), and epicatechin gallate (607.84 ± 22.65 μg/mL). The cell viabilities of 0.5 % (0.0875 mg/mL) green tea, the noncytotoxic concentration, were 96.04 ± 6.76 and 93.44 ± 0.95 % in B16F10 melanoma cells and human skin fibroblasts, respectively. Green tea exhibited the activities against skin aging, including the significant suppression of melanin production via inhibition of tyrosinase and tyrosinase-related protein-2 activities, the potent antioxidant, and the significant matrix metalloproteinase-2 inhibition (p < 0.001). ConclusionsThe study results have shown that green tea may be a functional processed plant which could be used as an anti-skin aging agent in natural remedies, including food, health, and cosmetic products.

Time-series transcriptomic analysis reveals novel gene modules that control theanine biosynthesis in tea plant (Camellia sinensis).

Theanine (thea) is a unique non-protein amino acid in tea plant (Camellia sinensis) and one of the most important small molecular compounds for tea quality and health effects. The molecular mechanism that maintains thea biosynthesis is not clear but may be reflected in complicated biological networks as other secondary metabolites in plants. We performed an integrative transcriptomic analysis of tea seedlings bud and leave over the time-course of ethylamine (EA) treatment that activated thea pathway. We identified 54 consistent differentially expressed genes (cDEGs, 25 upregulated and 29 downregulated) during thea activation. Gene Ontology (GO) functional enrichment analysis of upregulated genes and downregulated genes showed that they may function as a cascade of biological events during their cooperative contribution to thea biosynthesis. Among the total cDEGs, a diversity of functional genes (e.g., enzymes, transcription factors, transport and binding proteins) were identified, indicating a hierarchy of gene control network underlying thea biosynthesis. A gene network associated with thea biosynthesis was modeled and three interconnected gene functional modules were identified. Among the gene modules, several topologically important genes (e.g., CsBCS-1, CsRP, CsABC2) were experimentally validated using a combined thea content and gene expression analysis. Collectively, we presented here for the first time a comprehensive landscape of the biosynthetic mechanism of thea controlled by a underling gene network, which might provide a theoretical basis for the identification of key genes that contribute to thea biosynthesis.

Comparative transcriptomic analysis of the tea plant (Camellia sinensis) reveals key genes involved in pistil deletion

BackgroundThe growth process of the tea plant (Camellia sinensis) includes vegetative growth and reproductive growth. The reproductive growth period is relatively long (approximately 1.5 years), during which a large number of nutrients are consumed, resulting in reduced tea yield and quality, accelerated aging, and shortened economic life of the tea plant. The formation of unisexual and sterile flowers can weaken the reproductive growth process of the tea plant. To further clarify the molecular mechanisms of pistil deletion in the tea plant, we investigated the transcriptome profiles in the pistil-deficient tea plant (CRQS), wild tea plant (WT), and cultivated tea plant (CT) by using RNA-Seq.ResultsA total of 3683 differentially expressed genes were observed between CRQS and WT flower buds, with 2064 upregulated and 1619 downregulated in the CRQS flower buds. These genes were mainly involved in the regulation of molecular function and biological processes. Ethylene synthesis–related ACC synthase genes were significantly upregulated and ACC oxidase genes were significantly downregulated. Further analysis revealed that one of the WIP transcription factors involved in ethylene synthesis was significantly upregulated. Moreover, AP1 and STK, genes related to flower development, were significantly upregulated and downregulated, respectively.ConclusionsThe transcriptome analysis indicated that the formation of flower buds with pistil deletion is a complex biological process. Our study identified ethylene synthesis, transcription factor WIP, and A and D-class genes, which warrant further investigation to understand the cause of pistil deletion in flower bud formation.

Authentication of Ginkgo biloba Herbal Products by a Novel Quantitative Real-Time PCR Approach.

Ginkgo biloba is a widely used medicinal plant. Due to its potential therapeutic effects, it is an ingredient in several herbal products, such as plant infusions and plant food supplements (PFS). Currently, ginkgo is one of the most popular botanicals used in PFS. Due to their popularity and high cost, ginkgo-containing products are prone to be fraudulently substituted by other plant species. Therefore, this work aimed at developing a method for G. biloba detection and quantification. A new internal transcribe spacer (ITS) marker was identified, allowing the development of a ginkgo-specific real-time polymerase chain reaction (PCR) assay targeting the ITS region, with high specificity and sensitivity, down to 0.02 pg of DNA. Additionally, a normalized real-time PCR approach using the delta cycle quantification (ΔCq) method was proposed for the effective quantification of ginkgo in plant mixtures. The method exhibited high performance parameters, namely PCR efficiency, coefficient of correlation and covered dynamic range (50–0.01%), achieving limits of detection and quantification of 0.01% (w/w) of ginkgo in tea plant (Camellia sinensis). The quantitative approach was successfully validated with blind mixtures and further applied to commercial ginkgo-containing herbal infusions. The estimated ginkgo contents of plant mixture samples suggest adulterations due to reduction or almost elimination of ginkgo. In this work, useful and robust tools were proposed to detect/quantify ginkgo in herbal products, which suggests the need for a more effective and stricter control of such products.

Chemical Composition of a Supercritical Fluid (Sfe-CO2) Extract from Baeckea frutescens L. Leaves and Its Bioactivity Against Two Pathogenic Fungi Isolated from the Tea Plant (Camellia sinensis (L.) O. Kuntze).

Colletotrichum gloeosporioides and Pseudopestalotiopsis camelliae-sinensis are the two most important tea plant (Camellia sinensis L.) pathogenic fungi. Interest in natural plant extracts as alternatives to synthetic chemical fungicides to control plant pathogens is growing. In this study, the volatile fraction of Baeckea frutescens L. was extracted by supercritical fluid extraction (SFE-CO2), and its chemical composition was analyzed, and investigated for its antifungal activity against C. gloeosporioides and P. camelliae. The major constituents of the volatile fraction were β-caryophyllene (28.05%), α-caryophyllene (24.02%), δ-cadinene (6.29%) and eucalyptol (5.46%) in B. frutescens SFE-CO2 extracts. The terpineol, linalool, terpinen-4-ol and eucalyptol showed strong contact antifungal activity against P. camelliae and C. gloeosporioides with median inhibitory concentration (MIC50) in the range of 0.69 μL/mL to 2.79 μL/mL and 0.62 μL/mL to 2.18 μL/mL, respectively. Additionally, the volatile fraction had high fumigation antifungal activity against P. camelliae and C. gloeosporioides with an inhibition rate between 20.87% and 92.91%. Terpineol presented the highest antifungal activity in the contact and fumigation toxicity assays. Terpineol, linalool, terpinen-4-ol and eucalyptol were associated with the most active chemical compounds in the volatile fraction against the fungi. The results suggest that B. frutescens SFE-CO2 extracts are potential ingredients to develop a natural fungicide for control of tea plant pathogens.

Regulation mechanisms of humic acid on Pb stress in tea plant (Camellia sinensis L.)

Though the interaction between humic acid (HA) and heavy metals has been widely reported, the effects of HA on the toxicity of heavy metals to plants are still in debate. In this study, the regulation mechanisms of HA on Pb stress in tea plant (Camellia sinensis L.) was investigated through hydroponic experiments, and the experimental results were explained by using transmission electron microscope (TEM), scanning transmission X-ray microscopes (STXM) and isobaric tags for relative and absolute quantitation (iTRAQ) differential proteomics. Significant alleviation of Pb stress was found with HA coexistence. TEM results showed that HA greatly mitigated the damage of cells caused by Pb stress. Compared with sole Pb treatment, the addition of HA increased the contents of pectin and pectic acid in the cell wall by 10.5% and 30.5%, while arabinose (Ara) and galactose (Gal) decreased by 20.5% and 15.9%, respectively, which were beneficial for increasing Pb adsorption capacity of the cell wall and promoting cell elongation. Moreover, iTRAQ differential proteomics analysis proved that HA strengthened the antioxidant system, promoted the synthesis of cell wall, and stabilized protein and sulfur-containing substance metabolism in molecular level. Notably, the concentration of calcium (Ca) in the cell wall of HA coexistence treatment was 47.4% higher than Pb treatment. STXM results also indicated that the distribution of Ca in the cell wall was restored with the presence of HA. This might promote the formation of the egg-box model, thus alleviating Pb stress in cells. Our results reveal the regulation mechanisms of HA on Pb detoxification in plants and provide useful information for improving the safety of agricultural products.

Physiological, biochemical and genetic responses of Caucasian tea (Camellia sinensis (L.) Kuntze) genotypes under cold and frost stress.

BackgroundCold and frost are two serious factors limiting the yield of many crops worldwide, including the tea plant (Camellia sinensis (L.) Kuntze). The acclimatization of tea plant from tropical to temperate climate regions resulted in unique germplasm in the North–Western Caucasus with extremely frost-tolerant genotypes.MethodsThe aim of the current research was to evaluate the physiological, biochemical and genetic responses of tolerant and sensitive tea cultivars exposed to cold (0 to +2 °C for 7 days) and frost (−6 to −8 °C for 5 days). Relative water content, cell membranes integrity, pH of the cell sap, water soluble protein, cations, sugars, amino acids were measured under cold and frost. Comparative expression of the following genes ICE1, CBF1, WRKY2, DHN1, DHN2, DHN3, NAC17, NAC26, NAC30, SnRK1.1, SnRK1.2, SnRK1.3, bHLH7, bHLH43, P5CS, LOX1, LOX6, LOX7 were analyzed.ResultsWe found elevated protein (by 3–4 times) and cations (potassium, calcium and magnesium) contents in the leaves of both cultivars under cold and frost treatments. Meanwhile, Leu, Met, Val, Thr, Ser were increased under cold and frost, however tolerant cv. Gruzinskii7 showed earlier accumulation of these amino acids. Out of 18 studied genes, 11 were expressed at greater level in the frost- tolerant cultivar comparing with frost-sensitive one: ICE1, CBF1, WRKY2, DHN2, NAC17, NAC26, SnRK1.1, SnRK1.3, bHLH43, P5CS and LOX6. Positive correlations between certain amino acids namely, Met, Thr, Leu and Ser and studied genes were found. Taken together, the revealed cold responses in Caucasian tea cultivars help better understanding of tea tolerance to low temperature stress and role of revealed metabolites need to be further evaluated in different tea genotypes.