Shoots Of Tea Plant Research Articles

Root-specific theanine metabolism and regulation at the single-cell level in tea plants (Camellia sinensis)

Root-synthesized secondary metabolites are critical quality-conferring compounds of foods, plant-derived medicines, and beverages. However, information at a single-cell level on root-specific secondary metabolism remains largely unexplored. L-Theanine, an important quality component of tea, is primarily synthesized in roots, from which it is then transported to new shoots of tea plant. In this study, we present a single-cell RNA sequencing (scRNA-seq)-derived map for the tea plant root, which enabled cell-type-specific analysis of glutamate and ethylamine (two precursors of theanine biosynthesis) metabolism, and theanine biosynthesis, storage, and transport. Our findings support a model in which the theanine biosynthesis pathway occurs via multicellular compartmentation and does not require high co-expression levels of transcription factors and their target genes within the same cell cluster. This study provides novel insights into theanine metabolism and regulation, at the single-cell level, and offers an example for studying root-specific secondary metabolism in other plant systems.

Physiological and metabolomic analyses reveal the resistance response mechanism to tea aphid infestation in new shoots of tea plants (Camellia sinensis)

The tea plant (Camellia sinensis (L.) O. Kuntze) is an important woody economic crop and often attacked by various insect pests such as tea aphids (Toxoptera aurantii Boyer de Fonscolombe). To reveal tea plant responses to tea aphid infestation, tea aphid-resistant cultivar ‘Taicha13’ and -susceptible one ‘Jingfeng’ were first screened, and then widely targeted metabolomics profiling, biochemical parameter and RT-qPCR analyses were performed in three samples, TCN (tea aphid non-infested ‘Taicha13’), JFN (tea aphid non-infested ‘Jingfeng’) and JFA (tea aphid-infested ‘Jingfeng’). The results showed amnio acid (plus proline) and protein content, and amnio acid: sucrose ratio were significantly less abundant in TCN relative to JFN, suggesting TCN is low nutritional quality for tea aphids. Metabolite profiling revealed that flavonoids and JA-isoleucine (JA-Ile) were more enriched in TCN and JFA compared to JFN, proposing effective roles of flavonoids and JA-Ile in tea plant defense against tea aphid infestation. RT-qPCR results demonstrated that five JA biosynthesis genes (CsLOX2, CsLOX3, CsOPR, CsAOC and CsJAR) in exception of CsAOS, and six flavonoid biosynthesis genes (CsPAL, CsC4H, CsCHS1, CsCHS2, CsFL and CsDFR) shared similarly higher abundance in TCN than in JFN, and exhibited elevated expression levels in JFA relative to JFN, further suggesting flavonoids and JA confer tea plant resistance against tea aphid feeding. Overall, our findings uncovered that tea aphid-infestation activated defensive responses and highlighted involvement of nutritional quality, flavonoids and JA-Ile in tea plant resistance against tea aphid infestation.

Zno nanoparticles: improving photosynthesis, shoot development, and phyllosphere microbiome composition in tea plants

BackgroundNanotechnology holds revolutionary potential in the field of agriculture, with zinc oxide nanoparticles (ZnO NPs) demonstrating advantages in promoting crop growth. Enhanced photosynthetic efficiency is closely linked to improved vigor and superior quality in tea plants, complemented by the beneficial role of phyllosphere microorganisms in maintaining plant health. However, the effects of ZnO NPs on the photosynthesis of tea plants, the sprouting of new shoots, and the community of phyllosphere microorganisms have not been fully investigated.ResultsThis study investigated the photosynthetic physiological parameters of tea plants under the influence of ZnO NPs, the content of key photosynthetic enzymes such as RubisCO, chlorophyll content, chlorophyll fluorescence parameters, transcriptomic and extensive targeted metabolomic profiles of leaves and new shoots, mineral element composition in these tissues, and the epiphytic and endophytic microbial communities within the phyllosphere. The results indicated that ZnO NPs could enhance the photosynthesis of tea plants, upregulate the expression of some genes related to photosynthesis, increase the accumulation of photosynthetic products, promote the development of new shoots, and alter the content of various mineral elements in the leaves and new shoots of tea plants. Furthermore, the application of ZnO NPs was observed to favorably influence the microbial community structure within the phyllosphere of tea plants. This shift in microbial community dynamics suggests a potential for ZnO NPs to contribute to plant health and productivity by modulating the phyllosphere microbiome.ConclusionThis study demonstrates that ZnO NPs have a positive impact on the photosynthesis of tea plants, the sprouting of new shoots, and the community of phyllosphere microorganisms, which can improve the growth condition of tea plants. These findings provide new scientific evidence for the application of ZnO NPs in sustainable agricultural development and contribute to advancing research in nanobiotechnology aimed at enhancing crop yield and quality.Graphical

Dynamic DNA Methylation Regulates Season-Dependent Secondary Metabolism in the New Shoots of Tea Plants.

Plant secondary metabolites are critical quality-conferring compositions of plant-derived beverages, medicines, and industrial materials. The accumulations of secondary metabolites are highly variable among seasons; however, the underlying regulatory mechanism remains unclear, especially in epigenetic regulation. Here, we used tea plants to explore an important epigenetic mark DNA methylation (5mC)-mediated regulation of plant secondary metabolism in different seasons. Multiple omics analyses were performed on spring and summer new shoots. The results showed that flavonoids and theanine metabolism dominated in the metabolic response to seasons in the new shoots. In summer new shoots, the genes encoding DNA methyltransferases and demethylases were up-regulated, and the global CG and CHG methylation reduced and CHH methylation increased. 5mC methylation in promoter and gene body regions influenced the seasonal response of gene expression; the amplitude of 5mC methylation was highly correlated with that of gene transcriptions. These differentially methylated genes included those encoding enzymes and transcription factors which play important roles in flavonoid and theanine metabolic pathways. The regulatory role of 5mC methylation was further verified by applying a DNA methylation inhibitor. These findings highlight that dynamic DNA methylation plays an important role in seasonal-dependent secondary metabolism and provide new insights for improving tea quality.

Haem Oxygenase 1 is a potential target for creating etiolated/albino tea plants (Camellia sinensis) with high theanine accumulation.

Theanine content is highly correlated with sensory quality and health benefits of tea infusion. The tender shoots of etiolated and albino tea plants contain higher theanine than the normal green tea plants and are valuable materials for high quality green tea processing. However, why these etiolated or albino tea plants can highly accumulate theanine is largely unknown. In this study, we observed an Arabidopsis etiolated mutant hy1-100 (mutation in Haem Oxygenase 1, HO1) that accumulated higher levels of glutamine (an analog of theanine). We therefore identified CsHO1 in tea plants and found CsHO1 is conserved in amino acid sequences and subcellular localization with its homologs in other plants. Importantly, CsHO1 expression in the new shoots was much lower in an etiolated tea plants 'Huangkui' and an albino tea plant 'Huangshan Baicha' than that in normal green tea plants. The expression levels of CsHO1 were negatively correlated with theanine contents in these green, etiolated and albino shoots. Moreover, CsHO1 expression levels in various organs and different time points were also negatively correlated with theanine accumulation. The hy1-100 was hypersensitive to high levels of theanine and accumulated more theanine under theanine feeding, and these phenotypes were rescued by the expression of CsHO1 in this mutant. Transient knockdown CsHO1 expression in the new shoots of tea plant using antisense oligonucleotides (asODN) increased theanine accumulation. Collectively, these results demonstrated CsHO1 negatively regulates theanine accumulation in tea plants, and that low expression CsHO1 likely contributes to the theanine accumulation in etiolated/albino tea plants.

CsGDH2.1 negatively regulates theanine accumulation in late-spring tea plants (Camellia sinensis var. sinensis).

Theanine, a unique and the most abundant non-proteinogenic amino acid in tea plants, endows tea infusion with the umami taste and anti-stress effects. Its content in tea correlates highly with green tea quality. Theanine content in new shoots of tea plants is high in mid-spring and greatly decreases in late spring. However, how the decrease is regulated is largely unknown. In a genetic screening, we observed that a yeast mutant, glutamate dehydrolase 2 (gdh2), was hypersensitive to 40mM theanine and accumulated more theanine. This result implied a role of CsGDH2s in theanine accumulation in tea plants. Therefore, we identified the two homologs of GDH2, CsGDH2.1 and CsGDH2.2, in tea plants. Yeast complementation assay showed that the expression of CsGDH2.1 in yeast gdh2 mutant rescued the theanine hypersensitivity and hyperaccumulation of this mutant. Subcellular localization and tissue-specific expression showed CsGDH2.1 localized in the mitochondria and highly expressed in young tissues. Importantly, CsGDH2.1 expression was low in early spring, and increased significantly in late spring, in the new shoots of tea plants. These results all support the idea that CsGDH2.1 regulates theanine accumulation in the new shoots. Moreover, the in vitro enzyme assay showed that CsGDH2.1 had glutamate catabolic activity, and knockdown of CsGDH2.1 expression increased glutamate and theanine accumulation in the new shoots of tea plants. These findings suggested that CsGDH2.1-mediated glutamate catabolism negatively regulates theanine accumulation in the new shoots in late spring, and provides a functional gene for improving late-spring green tea quality.

Feedback Inhibition Might Dominate the Accumulation Pattern of BR in the New Shoots of Tea Plants (Camellia sinensis).

Brassinosteroid (BR), a kind of polyhydroxylated steroid hormone, plays an important role in physiological and biochemical processes in plants. Studies were mainly focused on BR signaling and its exogenous spraying to help enhance crop yields. Few research studies are centered on the accumulation pattern of BR and its mechanism. Yet, it is crucial to unlock the mystery of the function of BR and its cross action with other hormones. Tea (Camellia sinensis (L.) O. Kuntze) is one of the important economic crops in some countries, and new shoots are the raw materials for the preparation of various tea products. Different concentrations of exogenous BR were reported to have different effects on growth and development. New shoots of tea plants can thus be considered a valuable research object to study the accumulation pattern of BR. In this study, the quantity of five BR components (brassinolide, 28-norbrassinolide, 28-homobrassinolide, castasterone, and 28-norcastasterone) in different tissues of tea plants, including buds (Bud), different maturity of leaves (L1, L2), and stems (S1, S2) were determined by UPLC-MS/MS. A total of 15 cDNA libraries of the same tissue with three repetitions for each were constructed and sequenced. The BR-accumulation pattern and gene expression pattern were combined together for weighted gene co-expression network analysis (WGCNA). BR-accumulation-relative genes were then screened using two methods, based on the K.in value and BR biosynthetic pathway (ko00905), respectively. The result showed that photosynthesis-related genes and CYP450 family genes were actively involved and might play important roles in BR accumulation and/or its accumulation pattern. First and foremost, feedback inhibition was more likely to dominate the accumulation pattern of BR in the new shoots of tea plants. Moreover, three conserved miRNAs with their target transcriptional factors and target mRNAs had been figured out from negative correlation modules that might be strongly linked to the BR-accumulation pattern. Our study provided an experimental basis for the role of BR in tea plants. The excavation of genes related to the accumulation pattern of BR provided the possibility of cross-action studies on the regulation of BR biosynthesis and the study between BR and other hormones.

Theanine Improves Salt Stress Tolerance via Modulating Redox Homeostasis in Tea Plants (Camellia sinensis L.).

Theanine, a unique non-proteinogenic amino acid, is one of the most abundant secondary metabolites in tea. Its content largely determines green tea quality and price. However, its physiological roles in tea plants remain largely unknown. Here, we showed that salt stress significantly increased the accumulation of glutamate, glutamine, alanine, proline, and γ-aminobutyric acid, as well as theanine, in the new shoots of tea plants. We further found that salt stress induced the expression of theanine biosynthetic genes, including CsGOGATs, CsAlaDC, and CsTSI, suggested that salt stress induced theanine biosynthesis. Importantly, applying theanine to the new shoots significantly enhanced the salt stress tolerance. Similar effects were also found in a model plant Arabidopsis. Notably, exogenous theanine application increased the antioxidant activity of the shoots under salt stress, suggested by reduced the reactive oxygen species accumulation and lipid peroxidation, as well as by the increased SOD, CAT, and APX activities and expression of the corresponding genes. Finally, genetic evidence supported that catalase-mediated antioxidant scavenging pathway is required for theanine-induced salt stress tolerance. Taken together, this study suggested that salt stress induces theanine biosynthesize in tea plants to enhance the salt stress tolerance through a CAT-dependent redox homeostasis pathway.

Beauveria bassiana: as a potential microbial biocontrol agent for tea mosquito bug, Helopeltis theivora Waterhouse (Hemiptera: Miridae) in Dooars and Darjeeling, India

BackgroundIn the present study, the efficacy of two isolates of Beauveria bassiana namely, BKN20 and BKN1/14 was evaluated against the tea mosquito bug (TMB), Helopeltis theivora, Waterhouse (Hemiptera: Miridae) damaging harvestable shoots of tea plants in the Dooars and Darjeeling regions of West Bengal, India.ResultsLaboratory study revealed that, in both isolates, BKN20 was more pathogenic than BKN1/14, exhibiting 76% mortality of the test insect. The BKN20 isolate was formulated as an aqueous suspension (5%AS), and evaluated against TMB through micro-plot trials in tea plantations. The Micro-plot field study revealed a maximum of 72.19% reduction in the shoot damage due to TMB in plots sprayed with a 1000 ml/ha concentration of BKN20 5%AS containing 2 × 107 conidia/ml, as compared to the synthetic insecticide (Thiamethoxam 25%WG), where a 63.12% reduction in the shoots was recorded. Furthermore, different concentrations of the formulated BKN20 5%AS were evaluated against the test insect at 2 locations of tea gardens in the Dooars and Darjeeling regions. The results from both locations revealed that 1000 and 1200 ml/ha concentrations of BKN20 5%AS (each concentration containing 2 × 107 conidia/ml) significantly (p < 0.05) reduced the TMB population and they were more effective than Thiamethoxam 25%WG (120 g/ha). However, non-significant differences in crop yields were recorded. The formulation BKN20 5%AS was found to be non-pathogenic to non-target insects, i.e. natural enemies present in the tea ecosystem. BKN20 5%AS had no phytotoxic effect on the tea leaves, with acceptable organoleptic attributes.ConclusionThe BKN20 isolate could be commercialized as an alternative microbial insecticide to reduce the load of chemical insecticides in the tea ecosystem.

Transcriptomic and proteomic analysis of putative digestive proteases in the salivary gland and gut of Empoasca (Matsumurasca) onukii Matsuda

BackgroundInfestation by tea green leafhoppers (Empoasca (Matsumurasca) onukii) can cause a series of biochemical changes in tea leaves. As a typical cell-rupture feeder, E. onukii secretes proteases while using its stylet to probe the tender shoots of tea plants (Camellia sinensis). This study identified and analyzed proteases expressed specifically in the salivary gland (SG) and gut of E. onukii through enzymatic activity assays complemented with an integrated analysis of transcriptomic and proteomic data.ResultsIn total, 129 contigs representing seven types of putative proteases were identified. Transcript abundance of digestive proteases and enzymatic activity assays showed that cathepsin B-like protease, cathepsin L-like protease, and serine proteases (trypsin- and chymotrypsin-like protease) were highly abundant in the gut but moderately abundant in the SG. The abundance pattern of digestive proteases in the SG and gut of E. onukii differed from that of other hemipterans, including Nilaparvata lugens, Laodelphax striatellus, Acyrthosiphum pisum, Halyomorpha halys and Nephotettix cincticeps. Phylogenetic analysis showed that aminopeptidase N-like proteins and serine proteases abundant in the SG or gut of hemipterans formed two distinct clusters.ConclusionsAltogether, this study provides insightful information on the digestive system of E. onukii. Compared to five other hemipteran species, we observed different patterns of proteases abundant in the SG and gut of E. onukii. These results will be beneficial in understanding the interaction between tea plants and E. onukii.

Cellulose accumulation in the secondary cell walls is negatively correlated with the tenderness of new shoots in Camellia sinensis.

Tea, which is usually made from the new shoots of tea plants (Camellia sinensis), is one of the most popular and healthy beverages. The tenderness of new shoots plays a critical role in the production of high-quality tea; however, the factors affecting the tenderness of the new shoots of tea have not been extensively studied. Here, we show that cellulose accumulation is negatively correlated with the tenderness of new shoots, including leaves and stems, by characterizing the cellulose content and visualizing cellulose in the new shoots of two cultivars ("Huangjinya" and "Yujinxiang") via toluidine blue staining. We further found that the number of cells with secondary cell walls (SCWs) and the thickness of SCWs increased with the maturity degree of stems in both cultivars. Because cellulose is the most abundant polymer in SCW, we next identified three cellulose synthase CsCesAs, CsCesA4, 7, and 8A, through homologous alignment with Arabidopsis AtCesAs. Protein sequence analysis revealed that these proteins were closely related based on the phylogenetic analysis. We finally showed that the gene expression of the three CsCesAs was positively correlated with the maturity degree and cellulose content of the new shoots. Thus, our data suggest that cellulose accumulation, especially in the SCWs, is negatively correlated with the tenderness of new shoots in tea plants.

Effect of partial pesticide spraying on the number of major pests and damage to new shoots of tea plants

Abstract We studied the effects of partial spraying, targeting tea [Camellia sinensis (L.) O. Kuntze] plants' plucking surfaces, on the annual number of major pests and damage to new shoots. The prevention of feeding damage to shoots caused by Empoasca onukii Matsuda (Hemiptera: Cicadellidae) was more stable when spraying at high volumes. However, the annual number of E. onukii with conventional spraying (200 L/1000 m2) was similar to the control (No pesticide). On the other hand, the prevention effect on shoots and the annual number of Scirtothrips dorsalis Hood (Thysanoptera: Thripidae) in partial spraying (40 L/1000 m2) was similar to conventional spraying. Moreover, although high volumes of spraying (1000 L/1000 m2) are usually needed to control Pseudaulacaspis pentagona (Targioni-Tozetti) (Hemiptera: Diaspididae), the male adult population of P. pentagona in the control and partially sprayed field tended to be lower than that in the conventional spraying. When using pesticides harmful to P. pentagona's natural enemies, partial spraying seems beneficial. Enough pesticide spraying to widely cover the leaf layer was thought necessary to protect the tea plants' new shoots. However, the spraying method targeting only the tea plants' plucking surfaces protects the new shoots with similar efficacy. Besides, partial spraying protects pests' natural enemies, allowing additional pest population control.

Transcriptomic analyses identify albino-associated genes of a novel albino tea germplasm \u2018Huabai 1\u2019

Albinism in shoots of tea plants is a common phenotypic expression which gives the tea infusion a pleasant umami taste. A novel natural albino mutant tea germplasm containing high amino acids content was found and named as ‘Huabai 1’. ‘Huabai 1’ has white jade tender shoots under low temperature and turns green with increased temperature. In order to understand the molecular mechanism of color change in leaf of ‘Huabai 1’, transcriptome analysis was performed to identify albino-associated differentially expressed genes (DEGs). A total of 483 DEGs were identified from white shoots of ‘Huabai 1’ compared to its green shoots. There were 15 DEGs identified to be involved in phenylpropanoid biosynthesis, which account for the majority of characterized DEGs. The metabolites related to phenylpropanoid biosynthesis revealed similar expression pattern of DEGs. Furthermore, metabolic pathways such as ubiquonone, tyrosine, and flavonoid biosynthesis associated with phenylpropanoid biosynthesis could also contribute to the color change in ‘Huabai 1’ tender shoots. Protein–protein interaction analysis revealed a hub protein NEDD8 (CSA009575) which interacted with many regulated genes in spliceosome, nitrogen metabolism, phenylpropanoid biosynthesis, and other pathways. In conclusion, the findings in this study indicate that the color change of ‘Huabai 1’ tender shoots is a combined effect of phenylpropanoid biosynthesis pathway and other metabolic pathways including flavonoid biosynthesis in tea plants. Chlorophyll biosynthesis-related genes LHCII and SGR may also play some roles in color change of ‘Huabai 1’.

Effectiveness of medium pruning on the reduction of radioactive cesium concentration in new shoots of tea plants in Ibaraki Prefecture, Japan

Tea (Camellia sinensis (L.) Kuntze) plantations in the northern part of Ibaraki Prefecture in Japan, approximately 100 km southwest of the Fukushima Daiichi Nuclear Power Station (FDNPS), were contaminated with radioactive cesium (Cs) released from the power station during the Fukushima Daiichi Nuclear Disaster in 2011. Radioactive Cs concentrations in new shoots were monitored over the 3 years since June 2011. The use of conventional skiffing management resulted in 64% lower radioactive Cs concentration in October 2011 compared with that in June 2011, and the concentration continued to decrease gradually thereafter. Medium pruning to approximately 40 cm from the ground decreased radioactive Cs concentrations in the first tea crop of 2012 and 2013 by 24–25% more than conventional skiffing management.

神奈川県産のチャ新芽における放射性セシウム濃度

神奈川県産のチャ新芽における放射性セシウム濃度

Secondary control effects of a new insecticide, pyrifluquinazon, applied on the mulberry scale, Pseudaulacaspis pentagona (Targioni), against the pests infesting new shoots of tea plant

Secondary control effects of a new insecticide, pyrifluquinazon, applied on the mulberry scale, Pseudaulacaspis pentagona (Targioni), against the pests infesting new shoots of tea plant

Effect of liming and seasonal variation on lead concentration of tea plant ( Camellia sinensis (L.) O. Kuntze)

Tea is a widely consumed beverage. However, recent studies revealed that there were an increasing number of cases of tea products exceeding the former maximum permissible concentration (MPC) in China for Pb (2 mg kg −1). Tea Pb contamination is an issue affecting trade and consumer confidence. Root uptake of Pb could contribute significantly to Pb accumulation in tea leaves due to the strong acidity of many tea garden soils. We conducted pot and field experiments to evaluate the effect of liming on Pb uptake by tea plants on two highly acidic soils (pH 3.6). Additions of CaCO 3 significantly increased soil pH by up to 1 unit and decreased soil extractable Pb by up to 32%. Liming resulted in a decrease in the proportion of Pb in the exchangeable and carbonate-bound fractions, with a concurrent increase in the fractions bound to Fe/Mn oxides and residues. Liming significantly decreased Pb concentrations of fine roots, stems and new shoots of tea plants in the pot experiment. In the field experiments, the effect of liming was not significant during the first year following CaCO 3 application, but became significant during the second and third years and Pb concentration in the new shoots was decreased by approximately 20–50%, indicating that liming of acidic tea garden soils is an effective way to reduce Pb contamination of tea. The study also reveals a distinct seasonal variation, with Pb concentration in the new shoots following the order of spring > autumn > summer.

Sequencing of cDNA clones and analysis of the expressed sequence tags (ESTs) of tea plant [Camellia sinensis (L.) O. Kuntze] young shoots

AbstractThe construction of two cDNA libraries from young shoots of tea plant [Camellia sinensis cv. Longjing 43 and Anji Baicha], the sequencing of Longjing 43 cDNA clones and the analysis of expressed sequence tags (ESTs) are reported. Totally, 4320 clones from the cDNA library of Longjing 43 were sequenced, and 2963 useful sequences were obtained, corresponding to 68.6% of clones. A total of 416 clones shorter than 150 bp and 863 repeated clones were excluded, the first 1684 valid tea plant ESTs were generated. Most of the ESTs were between 300 and 700 bp, with an average of 478 bp. Six hundred and seven ESTs with known function or putative function were identified by BlastN searches against the National Center of Biotechnology Institute (NCBI) non-redundant nucleotide databases, corresponding to more than 130 functional genes in the tea plant. The rest, 1077 ESTs, were novel gene partial or full sequences. The results indicated that EST sequencing was a rapid and effective approach to identification of novel functional genes for tea plant.

Seasonal variations in biosynthetic capacity for the synthesis of caffeine in tea leaves

Abstract Seasonal variations in the metabolic fate of adenine nucleotides prelabelled with [8-14C]adenine were examined in leaf disks prepared at one-month intervals, over the course of one year, from the shoots of tea plants (Camellia sinensis L. cv Yabukita) which were growing under natural field conditions. Incorporation of radioactivity into nucleic acids and catabolites of purine nucleotides was found throughout the experimental period, but incorporation into theobromine and caffeine was found only in the young leaves harvested from April to June. Methylation of xanthosine, 7-methylxanthine and theobromine was catalysed by gel-filtered leaf extracts from young shoots (April to June), but the reactions could not be detected in extracts from leaves in which no synthesis of caffeine was observed in vivo. By contrast, the activity of 5-phosphoribosyl-1-pyrophosphate synthetase was still found in leaves harvested in July and August.