Characterizing microRNAs and their targets in different organs of Camellia sinensis var. assamica.

To comprehensively annotate miRNAs and their targets in tea plant, Camellia sinensis, we sequenced small and messenger RNAs of 9 samples of Camellia sinensis var. assamica (YK-10), a diploid elite cultivar widely grown in southwest China. In order to identify targets of miRNAs, we sequenced two degradome sequencing profiles from leaves and roots of YK-10, respectively. By analyzing the small RNA-Seq profiles, we newly identified 137 conserved miRNAs and 23 species specific miRNAs in the genome of YK-10, which significantly improved the annotation of miRNAs in tea plant. Approximately 2000 differently expressed genes were identified when comparing RNA-Seq profiles of any two of the three organs selected in the study. Totally, more than 5000 targets of conserved miRNAs were identified in the two degradome profiles. Furthermore, our results suggest that a few miRNAs play roles in the biosynthesis pathways of theanine, caffeine and flavonoid. These results enhance our understanding of small RNA guided gene regulations in different organs of tea plant.

Genome-wide identification, characterization, and expression analysis of nucleotide-binding leucine-rich repeats gene family under environmental stresses in tea (Camellia sinensis).

Plants often use nucleotide-binding leucine-rich repeats (NLRs) to recognize specific virulence proteins and activate the hypersensitive response thereby defending against invaders. However, data on NLRs and the resistance mechanism of NLR protein mediation in tea plant are extremely limited. In this study, 400 and 303 CsNLRs were identified from the genomes of C. sinensis var. sinensis (CSS) and C. sinensis var. assamica (CSA), respectively. Phylogenetic analysis revealed that the numbers in CNL groups are predominant in both CSS and CSA. RNA-Seq revealed that the expression of CsNLRs is induced by Colletotrichum fructicola, cold, drought, salt stress and exogenous methyl jasmonate. The 21 CsCNLs that are highly expressed in tea plant under biotic and abiotic stresses as well as during bud dormancy and in different tissues are identified. Gene structure analysis revealed several cis-regulatory elements associated with phytohormones and light responsiveness in the promoter regions of these 21 CsCNLs.

Identification of tea based on CARS-SWR variable optimization of visible/near-infrared spectrum.

BACKGROUND: The identification of tea varieties is essential to obtain high-quality tea that can command a high price. To identify tea varieties quickly and non-destructively, and to fight against counterfeit and inferior products in the tea market, a new method of visible / near-infrared spectrum processing based on competitive adaptive reweighting algorithms-stepwise regression analysis (CARS-SWR) variable optimization is proposed in this paper. RESULTS: The spectral data of five different tea varieties were obtained by visible / near-infrared spectrometry. The spectral data were preprocessed by the multivariate scattering correction (MSC) algorithm. First, 20 wavelength variables were selected by CARS, and then six optimal wavelength variables were selected using the SWR method, based on the CARS optimal variables. The generalized regression neural network (GRNN) classification model and probabilistic neural network (PNN) classification model were established, based on spectral information from the full wavelength, the CARS preferred wavelength variable, the SWR preferred wavelength variable, and the CARS-SWR preferred wavelength variable. CONCLUSION: It was found that the CARS-SWR-PNN model had the best classification effect by comparing different modeling results. The classification accuracy of its training set and test set reached 100%. This shows that the CARS-SWR preferred variable method combined with the visible / near-infrared spectrum is feasible for the rapid and non-destructive identification of tea varieties. © 2019 Society of Chemical Industry.

Rapid detection of quality index of postharvest fresh tea leaves using hyperspectral imaging.

BACKGROUND: The quality of fresh tea leaves after harvest determines, to some extent, the quality and price of commercial tea. A fast and accurate method to evaluate the quality of fresh tea leaves is required. RESULTS: In this study, the potential of hyperspectral imaging in the range of 328-1115 nm for the rapid prediction of moisture, total nitrogen, crude fiber contents, and quality index value was investigated. Ninety samples of eight tea-leaf varieties and two picking standards were tested. Quantitative partial least squares regression (PLSR) models were established using a full spectrum, whereas multiple linear regression (MLR) models were developed using characteristic wavelengths selected by a successive projections algorithm (SPA) and competitive adaptive reweighted sampling. The results showed that the optimal SPA-MLR models for moisture, total nitrogen, crude fiber contents, and quality index value yielded optimal performance with coefficients of determination for prediction (R2 p) of 0.9357, 0.8543, 0.8188, 0.9168; root mean square error of 0.3437, 0.1097, 0.3795, 1.0358; and residual prediction deviation of 4.00, 2.56, 2.31, and 3.51, respectively. CONCLUSION: The results suggested that the hyperspectral imaging technique coupled with chemometrics was a promising tool for the rapid and nondestructive measurement of tea-leaf quality, and had the potential to develop multispectral imaging systems for future online detection of tea-leaf quality. © 2020 Society of Chemical Industry.

Use of mineral multi-elemental analysis to authenticate geographical origin of different cultivars of tea in Guizhou, China.

BACKGROUND: The geographical origin of tea (Camellia sinensis) can be traced using mineral elements in its leaves as fingerprints. However, the role that could be played by soil mineral elements in the geographical authentication of tea leaves has been unclear. In this study, 22 mineral elements in 73 pairs of tea leaves and soils from three regions (Pu'an, Duyun, and Liping) in Guizhou, China, were determined using inductively coupled plasma mass spectrometry (ICP-MS) and inductively coupled plasma atomic emission spectrometry (ICP-AES). The mineral element concentrations were processed by multivariate statistical analysis, including one-way analysis of variance (ANOVA), correlation analysis, principal component analysis (PCA), and stepwise linear discriminant analysis (S-LDA). RESULTS: Based on a one-way ANOVA, tea leaves and soils with different origins possessed unique mineral element fingerprints. Sixteen mineral element concentrations in tea leaves were significantly correlated with those in soils (P < 0.05). The geographical origins of tea leaves were effectively differentiated using the 16 correlated mineral elements combined with PCA. The S-LDA model offered a 100% differentiation rate, and six indicative elements (phosphorus, Sr, U, Pb, Cd, and Cr) were selected as important fingerprinting markers for the geographic traceability of tea leaves. The accurate discrimination rate of geographical origin was unaffected by the cultivars of tea in the S-LDA model. CONCLUSIONS: Mineral elements in soils played an important role in the geographical authentication of tea leaves. Mineral elemental concentrations with significant correlations between tea leaves and soils could be robust, and could be used to trace the geographical origins of tea leaves. © 2020 Society of Chemical Industry.

Synchronous fluorescence spectroscopy combined with chemometrics for determination of total phenolic content and antioxidant activity in different tea types.

BACKGROUND: The aim of this study is to monitor the antioxidant activity (AA) and total phenolic content (TPC) variations of different kinds of teas depending on the infusion temperature and time by using synchronous fluorescence spectroscopy (SFS) combined chemometrics as a rapid method. In this study, black tea, oolong tea, green tea, and green tea powder (matcha) samples were brewed at 80 °C and 96 °C for 2.5 to 30 min. Synchronous fluorescence spectra were recorded at different wavelength interval (Δλ) values for optimal models. AA and TPC of tea samples were determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay and Folin-Ciocalteu methods as reference methods, respectively. Partial least square (PLS) method was used for correlation between reference methods and SFS method. RESULTS: Results showed that SFS combined with chemometrics could be an alternative rapid way to monitor TPC and AA of teas with 0.932 and 0.918 of validation R2 values in fermented teas (black tea-oolong) while with 0.961 and 0.860 of validation R2 values in non-fermented teas (green tea and green tea powder), respectively. Limit of detection (LOD) and root mean square error of prediction (RMSEP) values were ≤ 6.61 µg mL-1 and ≤ 17.42, respectively. CONCLUSION: Based on the lowest R2 value (0.860) on TPC analysis, the proposed method is more appropriate for AA analysis of green tea and green tea powder. Furthermore, infusion time was more effective for obtaining different amounts of TPC and AA in fermented tea types while only infusion temperature was effective on green tea and green tea powder samples. Therefore, obtained calibration-validation models gave better results for fermented tea types.

Characterization of genome-wide genetic variations between two varieties of tea plant (Camellia sinensis) and development of InDel markers for genetic research.

BACKGROUND: Single nucleotide polymorphisms (SNPs) and insertions/deletions (InDels) are the major genetic variations and are distributed extensively across the whole plant genome. However, few studies of these variations have been conducted in the long-lived perennial tea plant. RESULTS: In this study, we investigated the genome-wide genetic variations between Camellia sinensis var. sinensis 'Shuchazao' and Camellia sinensis var. assamica 'Yunkang 10', identified 7,511,731 SNPs and 255,218 InDels based on their whole genome sequences, and we subsequently analyzed their distinct types and distribution patterns. A total of 48 InDel markers that yielded polymorphic and unambiguous fragments were developed when screening six tea cultivars. These markers were further deployed on 46 tea cultivars for transferability and genetic diversity analysis, exhibiting information with an average 4.02 of the number of alleles (Na) and 0.457 of polymorphism information content (PIC). The dendrogram showed that the phylogenetic relationships among these tea cultivars are highly consistent with their genetic backgrounds or original places. Interestingly, we observed that the catechin/caffeine contents between 'Shuchazao' and 'Yunkang 10' were significantly different, and a large number of SNPs/InDels were identified within catechin/caffeine biosynthesis-related genes. CONCLUSION: The identified genome-wide genetic variations and newly-developed InDel markers will provide a valuable resource for tea plant genetic and genomic studies, especially the SNPs/InDels within catechin/caffeine biosynthesis-related genes, which may serve as pivotal candidates for elucidating the molecular mechanism governing catechin/caffeine biosynthesis.

De novo transcriptome and phytochemical analyses reveal differentially expressed genes and characteristic secondary metabolites in the original oolong tea (Camellia sinensis) cultivar 'Tieguanyin' compared with cultivar 'Benshan'.

BACKGROUND: The two original plants of the oolong tea cultivar ('Tieguanyin') are "Wei shuo" 'Tieguanyin'-TGY (Wei) and "Wang shuo" 'Tieguanyin'-TGY (Wang). Another cultivar, 'Benshan' (BS), is similar to TGY in its aroma, taste, and genetic make-up, but it lacks the "Yin Rhyme" flavor. We aimed to identify differences in biochemical characteristics and gene expression among these tea plants. RESULTS: The results of spectrophotometric, high performance liquid chromatography (HPLC), and gas chromatography-mass spectrometry (GC-MS) analyses revealed that TGY (Wei) and TGY (Wang) had deeper purple-colored leaves and higher contents of anthocyanin, catechins, caffeine, and limonene compared with BS. Analyses of transcriptome data revealed 12,420 differentially expressed genes (DEGs) among the cultivars. According to a Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis, the flavonoid, caffeine, and limonene metabolic pathways were highly enriched. The transcript levels of the genes involved in these three metabolic pathways were not significantly different between TGY (Wei) and TGY (Wang), except for two unigenes encoding IMPDH and SAMS, which are involved in caffeine metabolism. The comparison of TGY vs. BS revealed eight up-regulated genes (PAL, C4H, CHS, F3'H, F3H, DFR, ANS, and ANR) and two down-regulated genes (FLS and CCR) in flavonoid metabolism, four up-regulated genes (AMPD, IMPDH, SAMS, and 5'-Nase) and one down-regulated XDH gene in caffeine metabolism; and two down-regulated genes (ALDH and HIBADH) in limonene degradation. In addition, the expression levels of the transcription factor (TF) PAP1 were significantly higher in TGY than in BS. Therefore, high accumulation of flavonoids, caffeine, and limonene metabolites and the expression patterns of their related genes in TGY might be beneficial for the formation of the "Yin Rhyme" flavor. CONCLUSIONS: Transcriptomic, HPLC, and GC-MS analyses of TGY (Wei), TGY (Wang), and BS indicated that the expression levels of genes related to secondary metabolism and high contents of catechins, anthocyanin, caffeine, and limonene may contribute to the formation of the "Yin Rhyme" flavor in TGY. These findings provide new insights into the relationship between the accumulation of secondary metabolites and sensory quality, and the molecular mechanisms underlying the formation of the unique flavor "Yin Rhyme" in TGY.

The suitability of rare earth elements for geographical traceability of tea leaves.

BACKGROUND: Rare earth elements (REEs) have been used for the identification of the geographical origins of an increasing number of foods. This study analyzed the effects of geographical origin, harvest season, variety, and their interactions on REEs in tea leaves to investigate whether REEs were suitable for geographical identification of tea leaves. Tea leaves of different varieties and the corresponding soils were collected in different seasons from different areas of China. The concentrations of 14 REEs in tea leaves and soils were determined, and then analyzed with one-way analysis of variance (ANOVA), multi-way ANOVA, correlation analysis, and linear discriminant analysis. RESULTS: All factors significantly affected the contents of REEs in tea leaves. The concentrations of REEs in tea leaves were related to those in provenance soils. However, the concentrations of most REEs in tea leaves were primarily affected by the harvest season. CONCLUSION: Seasonal variations should be considered when REE fingerprinting is applied for the identification of tea for authentication purposes. © 2019 Society of Chemical Industry.

Geographical origin traceability of Keemun black tea based on its non-volatile composition combined with chemometrics.

BACKGROUND: Non-volatile compounds play a key role in the quality and price of Keemun black tea (KBT). The non-volatile compounds in KBT samples from different producing areas normally vary greatly. The development of rapid methods for tracing the geographical origin of KBT is useful. In this study, we develop models for the discrimination of KBT's geographical origin based on non-volatile compounds. RESULTS: Seventy-two KBT samples were collected from five towns in Anhui province to determine 13 KBT compounds by high-performance liquid chromatography (HPLC). Analysis of variance showed that the content of 13 compounds in KBT indicated significant differences (P < 0.05) among five towns. Three multivariate statistical models including principal component analysis (PCA), soft independent modeling of class analogy (SIMCA), and linear discriminant analysis (LDA) were built to discriminate origin. Principal component analysis effectively extracted three principal components, namely theaflavins, galloylated catechins, and simple catechins. The high sensitivity (64.5%-99.2%) was achieved of SIMCA model. To establish the discriminant functions, six variables (gallic acid, (+)-catechin, (-)-epigallocatechin gallate, theaflavin-3-gallate, theaflavin-3,3'-di-gallate, and total theaflavins) were chosen from 13 variables, and LDA was applied. This gave a satisfactory overall correct classification rate (94.4%) and cross-validation rate (88.9%) for KBT samples. CONCLUSION: The results showed that HPLC analysis together with chemometrics is a reliable approach for tracing KBT and guaranteeing its authenticity. © 2019 Society of Chemical Industry.

Transcriptome-wide identification and expression profile analysis of the bHLH family genes in Camellia sinensis.

The tea plant is an important commercial horticulture crop cultivated worldwide. Yield and quality of this plant are influenced by abiotic stress. The bHLH family transcription factors play a pivotal role in the growth and development, including abiotic stress response, of plants. A growing number of bHLH proteins have been functionally characterized in plants. However, few studies have focused on the bHLH proteins in tea plants. In this study, 120 CsbHLH TFs were identified from tea plants using computational prediction method. Structural analysis detected 23 conservative residues, with over 50% identities in the bHLH domain. Moreover, 103 CsbHLH proteins were assumed to bind DNA and encompassed 98 E-Box binders and 85 G-Box binders. The CsbHLH proteins were grouped into 20 subfamilies based on phylogenetic analysis and a previous classification system. A survey of transcriptome profiling screened 22 and 39 CsbHLH genes that were upregulated under heat and drought stress. Nine CsbHLH genes were validated using qRT-PCR. Results were approximately in accordance with transcriptome data. These genes could be induced by one or more abiotic stresses.

Genome-Wide Identification, Classification and Expression Analysis of the HSP Gene Superfamily in Tea Plant (Camellia sinensis).

Heat shock proteins (HSPs) function as molecular chaperones. These proteins are encoded by a multigene family whose members play crucial roles in plant growth, development and stress response. However, little is known about the HSP gene superfamily in tea plant. In this study, a total of 47 CsHSP genes were identified, including 7 CsHSP90, 18 CsHSP70, and 22 CssHSP genes. Phylogenetic and composition analyses showed that CsHSP proteins in the same subfamily have similar gene structures and conserved motifs, but significant differences exist in the different subfamilies. In addition, expression analysis revealed that almost all CsHSP genes were specifically expressed in one or more tissues, and significantly induced under heat and drought stress, implying that CsHSP genes play important roles in tea plant growth, development, and response to heat and drought stress. Furthermore, a potential interaction network dominated by CsHSPs, including HSP70/HSP90 organizing protein (HOP) and heat shock transcription factor (HSF), is closely related to the abovementioned processes. These results increase our understanding of CsHSP genes and their roles in tea plant, and thus, this study could contribute to the cloning and functional analysis of CsHSP genes and their encoded proteins in the future.

Metabolomic analysis reveals the composition differences in 13 Chinese tea cultivars of different manufacturing suitabilities.

BACKGROUND: Green tea and black tea are manufactured using appropriate tea cultivars in China. However, the metabolite differences relating to the manufacturing suitability of tea cultivars are unclear. In the present study, we performed a non-targeted metabolomic analysis on 13 Chinese tea cultivars using ultra-high performance liquid chromatography coupled with quadrupole-time-of-flight mass spectrometry to investigate comprehensively the metabolite differences between cultivars suitable for manufacturing green tea (GT cultivars) and cultivars suitable for manufacturing both green tea and black tea (G&BT cultivars). RESULTS: Multivariate statistical analysis and cluster analysis divided the 13 cultivars into two groups, namely GT cultivars and G&BT cultivars, which correlated with their manufacturing suitability. The GT cultivars contained higher levels of flavonoid glycosides, whereas the G&BT cultivars contained higher levels of catechins, dimeric catechins, phenolic acids and alkaloids. CONCLUSION: Metabolic pathway analysis revealed that the flavonoid pathway inclined toward the synthesis of flavonoid glycosides in GT cultivars, whereas it inclined toward the synthesis of catechins and phenolic acids in G&BT cultivars. The results of the present study will be helpful for discriminating the manufacturing suitability of tea cultivars and investigating their breeding. © 2017 Society of Chemical Industry.

Transcriptome analysis reveals self-incompatibility in the tea plant (Camellia sinensis) might be under gametophytic control.

BACKGROUND: Self-incompatibility (SI) is under genetic control and prevents inbreeding depression in angiosperms. SI mechanisms are quite complicated and still poorly understood in many plants. Tea (Camellia sinensis L.) belonging to the family of Theaceae, exhibits high levels of SI and high heterozygosity. Uncovering the molecular basis of SI of the tea plant may enhance breeding and simplify genomics research for the whole family. RESULTS: The growth of pollen tubes following selfing and crossing was observed using fluorescence microscopy. Self-pollen tubes grew slower than cross treatments from 24 h to 72 h after pollination. RNA-seq was employed to explore the molecular mechanisms of SI and to identify SI-related genes in C. sinensis. Self and cross-pollinated styles were collected at 24 h, 48 h and 72 h after pollination. Six RNA-seq libraries (SP24, SP48, SP72, CP24 CP48 and CP72; SP = self-pollinated, CP = cross-pollinated) were constructed and separately sequenced. In total, 299.327 million raw reads were generated. Following assembly, 63,762 unigenes were identified, and 27,264 (42.76 %) unigenes were annotated in five public databases: NR, KOG, KEGG, Swiss-Port and GO. To identify SI-related genes, the fragments per kb per million mapped reads (FPKM) values of each unigene were evaluated. Comparisons of CP24 vs. SP24, CP48 vs. SP48 and CP72 vs. SP72 revealed differential expression of 3,182, 3,575 and 3,709 genes, respectively. Consequently, several ubiquitin-mediated proteolysis, Ca(2+) signaling, apoptosis and defense-associated genes were obtained. The temporal expression pattern of genes following CP and SP was analyzed; 6 peroxidase, 1 polyphenol oxidase and 7 salicylic acid biosynthetic process-related genes were identified. The RNA-seq data were validated by qRT-PCR of 15 unigenes. Finally, a unigene (CL25983Contig1) with strong homology to the S-RNase was analyzed. It was mainly expressed in styles, with dramatically higher expression in self-pollinated versus cross-pollinated tissues at 24 h post-pollination. CONCLUSIONS: The present study reports the transcriptome of styles after cross- and self-pollination in tea and offers novel insights into the molecular mechanism behind SI in C. sinensis. We believe that this RNA-seq dataset will be useful for improvement in C. sinensis as well as other plants in the Theaceae family.

Transcriptomic analysis of Camellia ptilophylla and identification of genes associated with flavonoid and caffeine biosynthesis.

Camellia ptilophylla, or cocoa tea, is naturally decaffeinated and its predominant catechins and purine alkaloids are trans-catechins and theobromine Regular tea [Camellia sinensis (L.) O. Ktze.] is evolutionarily close to cocoa tea and produces cis-catechins and caffeine. Here, the transcriptome of C. ptilophylla was sequenced using the 101-bp paired-end technique. The quality of the raw data was assessed to yield 70,227,953 cleaned reads totaling 7.09 Gbp, which were assembled de novo into 56,695 unique transcripts and then clustered into 44,749 unigenes. In catechin biosynthesis, leucoanthocyanidin reductase (LAR) catalyzes the transition of leucoanthocyanidin to trans-catechins, while anthocyanidin synthase (ANS) and anthocyanidin reductase (ANR) catalyze cis-catechin production. Our data demonstrate that two LAR genes (CpLAR1 and CpLAR2) by C. ptilophylla may be advantageous due to the combined effects of this quantitative trait, permitting increased leucoanthocyanidin consumption for the synthesis of trans-catechins. In contrast, the only ANS gene observed in C. sinensis (CsANS) shared high identity (99.2%) to one homolog from C. ptilophylla (CpANS1), but lower identity (~80%) to another (CpANS2). We hypothesized that the diverged CpANS2 might have lost its ability to synthesize cis-catechins. C. ptilophylla and C. sinensis each contain two copies of ANR, which share high identity and may share the same function. Transcriptomic sequencing captured two N-methyl nucleosidase genes named NMT1 and NMT2. NMT2 was highly identical to three orthologous genes TCS2, PCS2, and ICS2, which did not undergo methylation in vitro; in contrast, NMT1 was less identical to TCS, PCS and ICS, indicating that NMT1 may undergo neofunctionalization.

Differing chemical compositions of three teas may explain their different effects on acute blood pressure in spontaneously hypertensive rats.

BACKGROUND: Heavy tea consumption is suggested to be unsuitable for hypertensive people. However, the bioactive substances in different varieties of tea leaves are very different. This study compares the effects of three Chinese teas - C. sinensis, C. ptilophylla and C. assamica var. kucha - on blood pressure (BP) and heart rate in spontaneously hypertensive rats (SHRs). RESULTS: Intragastric administration of C. sinensis extract led to an acute increase in systolic blood pressure (SBP), diastolic blood pressure (DBP) and heart rate in SHRs. However, C. ptilophylla and C. assamica var. kucha exerted no obvious influences on SBP, DBP or heart rate. Similar to the extract of C. sinensis, intragastric administration of caffeine also led to an acute increase in BP and heart rate in SHRs. In contrast, theobromine and theacrine - purine alkaloids predominantly contained in C. ptilophylla and C. assamica var. kucha, respectively - had no pressor effects. The effect of caffeine on BP was related to the regulation of plasma epinephrine and norepinephrine levels in SHRs. CONCLUSION: The different effects of C. sinensis, C. ptilophylla and C. assamica var. kucha on BP might be explained, at least partially, by the differences in the varieties and contents of purine alkaloids.

Antimicrobial activity of different tea varieties available in Pakistan.

In this antimicrobial study, various extracts of Green and Black tea (Camellia sinensis) and Lemon grass (Cymbopogon citrates) were evaluated for antimicrobial activities against six bacterial strains including both human pathogenic bacteria (Escherichia coli, Pseudomonas aeuroginosa, Staphylococcus aureus and Salmonella typhi) and plant pathogenic bacteria (Erwinia carotovora, Agro bacterium tumifaciens) and one fungal strain Candida albicans by disc diffusion susceptibility method. Of human pathogens, P. aeruginosa was most susceptible to all three different tea varieties; though rest of the strains also demonstrated prominent sensitivity. In comparison, black tea extracts were less activities than green tea and lemon grass. However, all the three tea varieties illustrated profound activity against plant pathogenic bacteria. Similarly, when extracts of tea were tested against C. albicans, green tea and lemon grass exhibited significant activity while black tea was mostly inactive.

Development and characterization of single nucleotide polymorphism markers in Camellia sinensis (Theaceae).

Tea is the second most popular non-alcoholic beverage in the world. In recent years, several molecular markers have been used in genetic studies of the tea plant. Yet, only a few single nucleotide polymorphisms (SNPs) have been reported. Here, we identified 818 putative SNPs from expressed sequence tag (EST) databases for the tea plant, which produced a frequency of 1 SNP/170 bp. A direct sequencing method was then used to verify 253 putative SNPs in genome DNA of 17 tea varieties. Fifty (20%) candidate and 299 new SNPs were identified. The haplotype polymorphism and nucleotide diversity of these markers ranged from 0 to 0.960 and 0 to 1.797%, respectively. Using these SNPs, the 17 varieties were classified into 2 groups by cluster analysis. The results indicate that Camellia sinensis-derived ESTs provide a valuable resource for SNP discovery. Furthermore, the abundance of SNPs in tea varieties is anticipated to generate the development of associated genetic studies, in addition to enhancing tea plant-breeding programs.

Classification and identification of tea diseases based on improved YOLOv7 model of MobileNeXt.

To address the issues of low accuracy and slow response speed in tea disease classification and identification, an improved YOLOv7 lightweight model was proposed in this study. The lightweight MobileNeXt was used as the backbone network to reduce computational load and enhance efficiency. Additionally, a dual-layer routing attention mechanism was introduced to enhance the model's ability to capture crucial details and textures in disease images, thereby improving accuracy. The SIoU loss function was employed to mitigate missed and erroneous judgments, resulting in improved recognition amidst complex image backgrounds.The revised model achieved precision, recall, and average precision of 93.5%, 89.9%, and 92.1%, respectively, representing increases of 4.5%, 1.9%, and 2.6% over the original model. Furthermore, the model's volum was reduced by 24.69M, the total param was reduced by 12.88M, while detection speed was increased by 24.41 frames per second. This enhanced model efficiently and accurately identifies tea disease types, offering the benefits of lower parameter count and faster detection, thereby establishing a robust foundation for tea disease monitoring and prevention efforts.

Analysis and classification of tea varieties using high-performance liquid chromatography and global retention models.

As a result of their metabolic processes, medicinal plants produce bioactive molecules with significant implications for human health, used directly for treatment or for pharmaceutical development. Chromatographic fingerprints with solvent gradients authenticate and categorise medicinal plants by capturing chemical diversity. This work focuses on optimising tea sample analysis in HPLC, using a model-based approach without requiring standards. Predicting the gradient profile effects on full signals was the basis to identify optimal separation conditions. Global models characterised retention and bandwidth for 14 peaks in the chromatograms across varied elution conditions, facilitating resolution optimisation of 63 peaks, covering 99.95 % of total peak area. The identified optimal gradient was applied to classify 40 samples representing six tea varieties. Matrices of baseline-corrected signals, elution bands, and band ratios, were evaluated to select the best dataset. Principal Component Analysis (PCA), k-means clustering, and Partial Least Squares-Discriminant Analysis (PLS-DA) assessed classification feasibility. Classification limitations were found reasonable due to tea processing complexities, involving drying and fermentation influenced by environmental conditions.