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Abstract Theanine (Thea) is a unique metabolite in tea plants, but its physiological functions remain elusive. A low soil pH increases cadmium (Cd) availability, affecting the quality of tea plant products. In this study, we found that Thea reversed the Cd-induced reduction in free amino acid (FAA) and caffeine (CAF) in the young tea leaves, as well as the down-regulation in the expression of nitrate transporters CsNRT1.2 and CsNRT2.5, and genes responsible for the nitrogen (N) assimilation. We demonstrated that Thea could alleviate Cd-induced oxidative stresses and enhance photosynthesis. Moreover, an ATP-binding cassette (ABC) transporter, CsABCG11.2, could uptake distinct Cd substrates and the five major amino acids in tea plants. Heterologous expression of CsABCG11.2 in yeast indicated a competitive absorption between Cd and Thea in a concentration-dependent pattern. CsABCG11.2-overexpressing Arabidopsis plants exhibited increased sensitivity to Cd due to enhanced Cd concentration, accumulation in the shoots, and reduction in the primary root length. Exogenous application of Thea at environmentally regular levels attenuated the adverse effects of Cd-induced growth inhibition and chlorosis in CsABCG11.2-overexpressing Arabidopsis plants. Knockdown of CsABCG11.2 tea plants significantly lowered Cd levels in young shoots. Our results suggest that Thea plays beneficial roles in alleviating Cd stress directly or indirectly by modulating CsABCG11.2-mediated Cd uptake and translocation within plants.

Tea tree is a fluorine (F)-enriched plant, leading to much concern about the safety of drinking tea from tea tree (Camellia sinensis (L.) Kuntze). Tea tree is a perennial leaf-harvested crop, and tea production in China is generally categorized as spring tea, summer tea and autumn tea in its annual growth rounds. However, the seasonally dynamic changes of F content and accumulation in the leaves and its drinking safety are poorly understood. In this study, 85 tea varieties cultivated under the same conditions were investigated to analyze the seasonal variation of F content and it’s relationships with F accumulation, aluminum (Al), calcium (Ca) and manganese (Mn) and hazard quotient (HQ) in young leaves (one bud and two leaves, YL) and mature leaves (canopy leaves, ML). The average F contents and accumulations were 350 mg kg−1 and 203 g ha−1 in YL, and they were 2451 mg kg−1 and 2578 g ha−1 in ML, respectively, with F mainly accumulated in ML. As the growing season progresses, the F content showed a gradual increase in YL, while a decrease in ML, inferring that F may be redistributed from mature leaves to young leaves. Additionally, the F content was quite different among tea varieties which are suitable for processing oolong tea, green tea, and black tea, with higher F accumulation in oolong tea varieties than in green and black tea varieties. Moreover, F content and accumulation could be obviously affected by the geographical origin of the tea tree varieties, with significantly higher F content in the varieties from F rich fluorite belts than other regions. Furthermore, F content and accumulation showed a significant positive correlation with the content of Al and Mn (p < 0.05). Based on a daily tea consumption of 8.7 g, the HQ was investigated to show that the proportion of tea leaves with HQ

Withering is one of the major processing steps critical for the quality of black tea. In this study, we investigated the mechanisms underlying the physicochemical changes in metabolites and gene expression during the withering process of black tea using metabolomic and transcriptomic approaches, respectively. Based on gas chromatography/mass spectrometry non-targeted metabolomic approaches (GC-MS) and ultra-high performance liquid chromatograph–tandem mass spectrometry (UHPLC-MS/MS), a total of 76 volatile compounds and 160 non-volatile compounds were identified from tea leaves, respectively. RNA-seq analysis revealed that the number of differentially expressed genes (DEGs) for the comparative combination of withering time (i.e., W4h, W6h, W8h, W10h, and W12h) compared with CK (i.e., fresh leaves) were 3634, 2906, 4127, 5736, and 7650, respectively. The core genes in starch metabolism, namely alpha-amylase (AMY) and beta-amylase (BAM), were upregulated as withering time increased. AMY and BAM contributed to the decomposition of starch to increase the soluble sugars. The content of tea leaf alcohols and aldehydes, which are the vital contributors for greenish aroma, gradually decreased as withering time increased due to the downregulation of associated genes while the compounds related to sweet and fruity characteristics increased due to the upregulated expression of related genes. Most DEGs involved in amino acids were significantly upregulated, leading to the increase in free amino acids content. However, DEGs involved in catechins metabolism were generally downregulated during withering, and resulted in a reduction in catechins content and the accumulation of theaflavins. The same trend was observed in alpha-linolenic acid metabolism-related genes that were downregulated and enhanced the reduction in grassy aroma in black tea. The weighted gene co-expression network analysis (WGCNA) of DEGs showed that one module can be associated with more components and one component can be regulated by various modules. Our findings provide new insights into the quality formation of black tea during the withering process.

In plants, the lysine and histidine transporter (LHT) family represent a class of proteins that mediate the uptake, translocation, and utilization of amino acids. The tea plant (Camellia sinensis) is a perennial evergreen with a relatively high level of amino acids. However, systematic identification and molecular characterization of the LHT gene family has rarely been reported in tea plants. In this study, 22 CsLHTs were identified from the ‘Shuchazao’ genome and classified into two groups. The modeled three-dimensional structure and the conserved domains presented a high similarity among the LHTs proteins. Moreover, it was predicted that a few genes were conserved through the analysis of the physiochemical characters, structures and cis-elements in promoters. The expression patterns in tea plants revealed that CsLHT7 was mainly expressed in the roots, and CsLHT4 and CsLHT11 exhibited relatively high expression in both the roots and leaves. Moreover, the expression of all three genes could be induced by organic nitrogen. Additionally, heterogeneous expression of CsLHT4, CsLHT7 and CsLHT11 in Arabidopsis thaliana decreased the aerial parts biomass compared with that in WT plants while significantly increased the rosette biomass only for CsLHT11 transgenic plants versus WT plants. Overall, our results provide fundamental information about CsLHTs and potential genes in N utilization for further analysis in tea plants.

Many attentions have been previously focused to identify the multiple biochemical components related to tea quality and health benefits, however, the natural variation of biochemical components present in tea germplasm has not been adequately evaluated. In this study, the main biochemical components, leaf morphological and yield characteristics were evaluated for four rounds of tea leaves in a panel of 87 elite tea cultivars suitable for black, green, or oolong tea. Significant variations were observed among the tea cultivars, as well as seasonal differences in the levels of the free amino acid (FAA), caffeine (CAF), tea polyphenols (TP), water extract (WE) and TP to FAA ratio (TP/FAA). Results showed that the average levels of FAA showed a seasonal change, with the highest level of 4.0% in the 1st spring tea in the cultivars suitable for green tea and the lowest of 3.2% in summer tea in the cultivars suitable for black tea. The average CAF content was highest 3.2% in the cultivars suitable for oolong tea in the 1st spring and the lowest 2.5% in the cultivars suitable for green tea in summer. Limited seasonal and varietal variations were noticed in the average levels of WE among the three categories of tea. In addition, significant natural variation of the morphological characteristics, bud length varying from 2.5 cm to 8.7 cm, bud density from 190.3 buds · m-2 to 1 730.3 buds · m-2, mature leaves biomass from 128.4 kg · hm-2 to 2 888.4 kg · hm-2, and yield component traits of 100 buds (one bud with two leaves) dry weight from 3.7 g to 37.7 g, tea yield/round from 444.6 kg · hm-2 to 905.3 kg · hm-2, were observed. The aim of our evaluation was not only to identify the advantages of seasonal and clonal variations but also to provide a new viewpoint for their further application. Representative accessions were selected from the germplasm to promote the establishment of an inherent biochemical constituent expressing the quality of black, green, and oolong tea. The findings might be utilized to establish early selection criteria to enhance the tea breeding and production program.

Surface-enhanced Raman scattering (SERS) provides an alternative rapid detection method for pesticide residues in food, but fenvalerate possesses poor affinity to the novel metal substrate, thus restricting its analysis. To break this bottleneck, a SERS-active platform with an Ag/rGO composite structure was engineered using a facile method for fenvalerate detection. Ag nanoparticles with a 60 nm diameter can grow evenly on the top and bottom of rGO layers under intense ultrasonic oscillation, and rGO in hybrid material acts as an ideal hotspot holder between the gaps of Ag nanoparticles, not only allowing the interaction area to be enhanced both electromagnetically and chemically but also enabling the capture and enrichment of fenvalerate pesticide molecules into the “hotspot” area to improve detection sensitivity. Ag/rGO composite substrate possesses superior SERS performance with an ultralow detectable concentration of 4-aminothiophenol (10−10 M) and good reproducibility, endowing the material with a better enhancement effect than pure Ag nanoparticles. When used as the SERS substrate for fenvalerate detection, Ag/rGO composite material showed excellent performance in both experiments and theoretical calculation, with the limit of detection (LOD) of fenvalerate being as low as 1.69 × 10−5 mg/kg and a detection model with an R2 of 99.2%, demonstrating its exciting potential as a SERS substrate for pesticides detection.

Nine tea cultivars planted in Enshi were selected and processed into “Lichuan black tea”. Sensory evaluation showed that cultivar had the greatest influence on taste and aroma quality, including sweetness, umami and concentration of taste, as well as sweet and floral fragrances of aroma. The non-volatile and volatile components were identified by UPLC-Q-TOF/MS and GC–MS, and PCA analysis showed good separation between cultivars, which could cause the difference in quality. Baiyaqilan, Meizhan and Echa 10 had a floral aroma, with obvious difference in their aromatic composition from other cultivars. Moreover, Echa 10 also had a strong sweet aroma. The key aroma components in Echa 10 (with the largest cultivation area) were further investigated by GC-O-MS combined with odor activity value (OAV) analysis, included β-damascenone, phenylethylaldehyde, nonenal, geraniol, linalool, jasmonone, (E)-2-nonenal, β-cyclocitral, (E)-β-ocimene, methyl salicylate, β-ionone, 2,6,10,10-tetramethyl-1-oxaspiro[4.5]dec-6-ene, citral, β-myrcene, nerol, phenethyl alcohol, benzaldehyde, hexanal, nonanoic acid, and jasmin lactone.

This study delved into the aroma characteristics of “Qingxiang” oolong tea, analyzing six different cultivars and their processing modes. The findings showed that both cultivars and processing modes have a significant impact on the oolong tea aroma system. The study identified 18 terpenoid volatiles (VTs), 11 amino-acid-derived volatiles (AADVs), 15 fatty-acid-derived volatiles (FADVs), 3 carotenoid-derived volatiles (CDVs), and 10 other compounds in oolong tea that differentiate it from green and black tea. The turn-over stage was found to be the primary processing stage for oolong tea aroma formation. Molecular sensory analysis revealed that the “fresh” odor attribute is the basis for its aroma, while “floral and fruity” fragrances are its aroma characteristics. The perception of oolong tea as “fresh” and “floral and fruity” is influenced by the interactions of its aroma components. These findings provide a new basis for breed improvement and process enhancement in oolong tea production.

This study fristly investigated the taste quality formation and leaf conducting tissue changes in six types of Chinese tea (green, black, oolong, yellow, white, and dark) made from Mingke No.1 variety. Non-targeted metabolomics showed the vital manufacturing processes (green tea-de-enzyming, black tea-fermenting, oolong tea-turning-over, yellow tea-yellowing, white tea-withering, and dark tea-pile-fermenting) were highly related to their unique taste formation, due to different fermentation degree in these processes. After drying, the retained phenolics, theanine, caffeine, and other substances significantly impacted each tea taste quality formation. Meanwhile, the tea leaf conducting tissue structure was significantly influenced by high processing temperature, and the change of its inner diameter was related to moisture loss during tea processing, as indicated by its significant different Raman characteristic peaks (mainly cellulose and lignin) in each key process. This study provides a reference for process optimization to improve tea quality.

Abstract Background The major aluminum (Al) detoxication mechanism of tea plant (Camellia sinensis), as an Al hyperaccumulator plant, is the fixation of almost 70% of Al in the cell walls. Pectin is the primary constituent of cell walls, a degree of methylation of pectin polysaccharides regulated by the pectin methylesterase (PME) genes can greatly affect the Al binding capacity. The knowledge on PME gene family in tea plant is still poor. Results We identified 66 (CsPME1-CsPME66) PME genes from C. sinensis genome. We studied their protein characterization, conserved motifs, gene structure, systematic evolution and gene expression under Al treatments, to establish a basis for in-depth research on the function of PMEs in tea plant. Gene structures analysis revealed that the majority of PME genes had 2–4 exons. Phylogenetic results pointed out that the PME genes from the same species displayed comparatively high sequence consistency and genetic similarity. Selective pressure investigation suggested that the Ka/Ks value for homologous genes of PME family was less than one. The expression of CsPMEs under three Al concentration treatments was tissue specific, eight PME genes in leaves and 15 in roots displayed a trend similar to of the Al contents and PME activities under Al concentration treatments, indicating that the degree of pectin de-esterification regulated by PME was crucial for Al tolerance of tea plant. Conclusions Sixty-six CsPME genes were identified for the first time in tea plant. The genome-wide identification, classification, evolutionary and transcription analyses of the PME gene family provided a new direction for further research on the function of PME gene in Al tolerance of tea plant.

A non-targeted metabolomics approach and sensory evaluation, coupled with multivariate statistical analysis, systematically uncover the impact of the rolling time on the quality parameters of black tea. GC-MS analysis reveals that a moderate extension of rolling time favorably contributes to the accumulation of characteristic aroma components in black tea. The volatile components reach their highest concentration in black tea samples processed during an 80-min rolling period. UHPLC-Q-TOF/MS analysis demonstrates a substantial decrease in the contents of catechins and flavonoids with an increase in rolling time. Simultaneously, the production of theaflavins, coupled with the degradation of green bitterness volatiles (GBVs), significantly contributes to the formation of endogenous aroma components in black tea. These findings underscore the close relationship between rolling time control and black tea quality, emphasizing that a moderate extension of the rolling time fosters the development of improved black tea flavor quality. The comprehensive quality evaluation indicates that the optimal duration is 80 min. However, the initial 0 to 20 min of rolling is a crucial phase for the genesis and transformation of black tea quality. This study offers valuable insights into the influence of rolling time on black tea quality, potentially enhancing future studies of rolling technology. It provides theoretical guidelines for optimizing the processing of Gongfu black tea.

Tea polysaccharide (TPS) is a bioactive compound extracted from tea. It has raised great interest among researchers due to its bioactivity. However, few studies focused on the diversity of TPS in its compositions and antioxidant activity. This study collected 140 different tea varieties from four tea germplasm gardens in China, and their TPSs in tea shoots were extracted. The extraction efficiency, composition contents, including neutral sugar, uronic acid, protein, and tea polyphenols, and the scavenging abilities of hydroxyl radical (·OH) and superoxide radical (O2-·) of 140 TPSs were determined and analyzed. The results showed significant differences in the compositions and antioxidant activities of TPS extracted from different tea varieties. By applying hierarchical clustering analysis (HCA), we selected nine tea varieties with high TPS extraction efficiency and 26 kinds of TPS with high antioxidant capacity.

Nitrogen (N) is an important contributor in regulating plant growth and development as well as secondary metabolites synthesis, so as to promote the formation of tea quality and flavor. Theanine, polyphenols, and caffeine are important secondary metabolites in tea plant. In this study, the responses of Camellia sinensis roots to N deprivation and resupply were investigated by metabolome and RNA-seq analysis. N deficiency induced content increase for most amino acids (AAs) and reduction for the remaining AAs, polyphenols, and caffeine. After N recovery, the decreased AAs and polyphenols showed a varying degree of recovery in content, but caffeine did not. Meanwhile, theanine increased in content, but its related synthetic genes were down-regulated, probably due to coordination of the whole N starvation regulatory network. Flavonoids-related pathways were relatively active following N stress according to KEGG enrichment analysis. Gene co-expression analysis revealed TCS2, AMT1;1, TAT2, TS, and GOGAT as key genes, and TFs like MYB, bHLH, and NAC were also actively involved in N stress responses in C. sinensis roots. These findings facilitate the understanding of the molecular mechanism of N regulation in tea roots and provide genetic reference for improving N use efficiency in tea plant.

Fresh tea leaves, both single bud and one leaf with a bud, were used as the test materials in this study. The variation in the main mechanical properties, such as texture and tensile properties, during the processing of Longjing tea was examined by using texture profile analysis (TPA) and stress–strain tensile tests. The plasticity showed a trend of first increasing and then decreasing during the processing, whereas the elasticity displayed the opposite tendency. The flexibility reached a maximum during the fixing stage and then slowly declined with a relatively small change. Initially, the maximum force dropped down and then gradually elevated later.Both the tensile strength and the fracture strain indicated an upward movement at the beginning and then a downfall afterward. The elastic modulus changed little before the final panning stage, then raised significantly. The correlation analysis revealed that the flexibility of tea leaves was highly positively correlated with water content. At water content of 30% and 50%, the plasticity and flexibility of tea leaves reached a clear peak and the maximum force was at a low level, which is suitable for the shaping of Longjing tea. The results also demonstrated that the main mechanical properties of different tender materials change differently during the processing. The research findings can provide parameters for optimizing the mechanical design and processing technology of Longjing tea.

In this study, the effects of different sun withering degrees (75% (CK), 69% (S69), 66% (S66), 63% (S63), and 60% (S60) water content in the withered leaves) on black tea sensory quality were investigated by means of sensory evaluation plus metabolomics analysis. Sensory evaluation results showed higher sensory quality scores for the black tea in S69–S66, due to better freshness, sweeter taste, and a sweet and even floral and fruity aroma. Additionally, 65 non-volatile components were identified using Ultra Performance Liquid Chromatography-Quadrupole-Time of Flight-Mass Spectrometry (UPLC-Q-TOF/MS). Among them, the content increase of amino acids and theaflavins was found to promote the freshness and sweetness of black tea. The aroma of tea was analyzed using combined Solvent Assisted Flavor Evaporation-Gas Chromatography-Mass Spectrometry (SAFE-GC-MS) and Headspace-Solid Phase Micro Extract-Gas Chromatography-Mass Spectrometry (HS-SPME-GC-MS), and 180 volatiles were identified, including 38 variable importance in projection (VIP) > 1 (p < 0.05) and 25 Odor Activity Value (OAV) > 1 volatiles. Statistical analysis revealed 11 volatiles as potential major aroma differential metabolites in black tea with a different sun withering degree, such as volatile terpenoids (linalool, geraniol, (E)-citral, and β-myrcene), amino-acid-derived volatiles (benzeneethanol, benzeneacetaldehyde, and methyl salicylate), carotenoid-derived volatiles (jasmone and β-damascenone), and fatty-acid-derived volatiles ((Z)-3-hexen-1-ol and (E)-2-hexenal). Among them, volatile terpenoids and amino acid derived volatiles mainly contributed to the floral and fruity aroma quality of sun-withered black tea.

The quality of traditional sunlight-dried orange black tea can be affected by weather variations, leading to its quality instability. This study investigated the feasibility of replacing sunlight drying with a new hot-air drying method in orange black tea production. The hot-air-dried orange black tea showed better sensory quality than the traditional outdoor-sunlight-dried tea, with a harmonious fruity aroma and sweet–mellow taste. The content of polyphenols and other quality components in the peel and tea leaves was significantly higher after hot-air drying than after sunlight drying. GC-MS analysis showed that the total number of volatile components of hot-air-dried tea (3103.46 μg/g) was higher than that of sunlight-dried tea (3019.19 μg/g). Compared with sunlight-dried orange black tea, the hot-air-dried orange black tea showed higher total antioxidant capacity, with an increase of 21.5% (FRAP), 7.5% (DPPH), and 17.4% (ABTS), as well as an increase of 38.1% and 36.3% in the inhibitory capacity on α-glucosidase and α-amylase activities. Further analysis of the effects of different drying temperatures (40, 45, 50, and 60 °C) on the quality of orange black tea showed that the tea quality gradually decreased with the increase in drying temperature, with the most obvious decrease in the quality of orange black tea at the drying temperature of 60 °C. Low-temperature (40 °C) dried tea had better aroma coordination, higher fruit flavor, greater sweet–mellow taste, and higher retention of functional active substances in orange peel and black tea. In summary, compared with traditional sunlight drying, the hot-air drying method could reduce the drying time from 90 h to 20 h and improve the sensory quality and functional activity of orange black tea, suggesting it can replace the traditional sunlight drying process. This work is significant for improving the quality of orange black tea in practical production.

Green tea processed by Echa 10 was shown to have a fresh and mellow taste as well as clean aroma with a clear honeysuckle fragrance. The colors of different Enshi green teas are closely related with the content of chlorophyll and chlorophyllide. The five green teas also vary in their aroma style. Echa 10 imparts a special honeysuckle fragrance, which was further analyzed by molecular sensory analysis and the formation of this honeysuckle fragrance was attributed to the key components of dodecane, octadecane, phenethyl alcohol, and jasmonone. In aroma evaluation, Echa 10 green tea showed the best performance, which is mainly related with the content of geraniol, linalool, phenethyl alcohol, and benzyl alcohol. Additionally, Echa 10 scored the highest in taste evaluation, which is mainly determined by the contents and ratios of tea polyphenols, amino acids, caffeine, and soluble sugars.

For tea plants, nitrogen (N) is a foundational element and large quantities of N are required during periods of roundly vigorous growth. However, the fluctuation of N in the tea garden could not always meet the dynamic demand of the tea plants. Autophagy, an intracellular degradation process for materials recycling in eukaryotes, plays an important role in nutrient remobilization upon stressful conditions and leaf senescence. Studies have proven that numerous autophagy-related genes (ATGs) are involved in N utilization efficiency in Arabidopsis thaliana and other species. Here, we identified an ATG gene, CsATG101, and characterized the potential functions in response to N in A. thaliana. The expression patterns of CsATG101 in four categories of aging gradient leaves among 24 tea cultivars indicated that autophagy mainly occurred in mature leaves at a relatively high level. Further, the in planta heterologous expression of CsATG101 in A. thaliana was employed to investigate the response of CsATG101 to low N stress. The results illustrated a delayed transition from vegetative to reproductive growth under normal N conditions, while premature senescence under N deficient conditions in transgenic plants vs. the wild type. The expression profiles of 12 AtATGs confirmed the autophagy process, especially in mature leaves of transgenic plants. Also, the relatively high expression levels for AtAAP1, AtLHT1, AtGLN1;1, and AtNIA1 in mature leaves illustrated that the mature leaves act as the source leaves in transgenic plants. Altogether, the findings demonstrated that CsATG101 is a candidate gene for improving annual fresh tea leaves yield under both deficient and sufficient N conditions via the autophagy process.

This paper investigated the effect and mechanism of theaflavins (TFs) on fluoride (F−) uptake and transport in the Caco-2 cell model through structural chemistry and transcriptome analysis. The results showed that the four major TFs (TF, TF3G, TF3′G and TFDG) at a 150 μg/mL concentration could all significantly decrease F− transport in Caco-2 cells after 2 h of treatment and, at 2 μg/mL F− concentration, the F− transport was more inclined to efflux. During transport, the F− retention in Caco-2 cells was significantly increased by TF3G while it was clearly decreased by TF. The interaction between TFs and F− was analyzed by Raman spectroscopy and isothermal titration calorimetry, and F− was shown to affect the π bond vibration on the benzene ring of TFs, thus influencing their stability. Additionally, F− showed weak binding to TF3G, TF3′G and TFDG, which may inhibit F− transport and absorption in the Caco-2 cell line. Transcriptome and RT-PCR analysis identified three key differentially expressed genes related to cell permeability, and TFs can be assumed to mediate F− transport by regulating the expression of permeability-related genes to change cell monolayer permeability and enhance cell barrier function; however, this needs to be further elucidated in future studies.

Steamed green tea has unique characteristics that differ from other green teas. However, the alteration patterns of non-volatile metabolites during steamed green tea processing are not fully understood. In this study, a widely targeted metabolomic method was employed to explore the changes in non-volatile metabolites during steamed green tea processing. A total of 735 non-volatile compounds were identified, covering 14 subclasses. Of these, 256 compounds showed significant changes in at least one processing step. Most amino acids, main catechins, caffeine, and main sugars were excluded from the analysis. The most significant alterations were observed during steaming, followed by shaping and drying. Steaming resulted in significant increases in the levels of most amino acids and their peptides, most phenolic acids, most organic acids, and most nucleotides and their derivates, as well as some flavonoids. Steaming also resulted in significant decreases in the levels of most lipids and some flavonoids. Shaping and drying caused significant increases in the levels of some flavonoids, phenolic acids, and lipids, and significant decreases in the levels of some amino acids and their peptides, some flavonoids, and some other compounds. Our study provides a comprehensive characterization of the dynamic alterations in non-volatile metabolites during steamed green tea manufacturing.

This study investigated the effect of different fermentation humidities (55%, 65%, 75%, 85% and 95%) on congou black tea quality and bioactivity. Fermentation humidity mainly affected the tea′s appearance, aroma and taste quality. The tea fermented at low humidity (75% or below) showed a decrease in tightness, evenness and moistening degree, as well as a heavy grassy and greenish scent, plus a green, astringent and bitter taste. The tea fermented at a high humidity (85% or above) presented a sweet and pure aroma, as well as a mellow taste, plus an increase of sweetness and umami. With increasing fermentation humidity, the tea exhibited a drop in the content of flavones, tea polyphenols, catechins (EGCG, ECG) and theaflavins (TF, TF-3-G), contrasted by a rise in the content of soluble sugars, thearubigins and theabrownins, contributing to the development of a sweet and mellow taste. Additionally, the tea showed a gradual increase in the total amount of volatile compounds and in the content of alcohols, alkanes, alkenes, aldehydes, ketones and acids. Moreover, the tea fermented at a low humidity had stronger antioxidant activity against 2, 2-Diphenyl-1-picrylhydrazyl (DPPH) and a higher inhibiting capability on the activities of α-amylase and α-glucosidase. Overall results indicated the desirable fermentation humidity of congou black tea should be 85% or above.

Peach leaf orange dark tea (ODT) is a fruity tea made by removing the pulp from peach leaf orange and placing dry Qingzhuan tea into the husk, followed by fixing them together and drying. Since the quality of traditional outdoor sunlight fixing (SL) is affected by weather instability, this study explored the feasibility of two new fixing methods, including hot air fixing (HA) and steam fixing (ST). Results showed that fixing method had a great impact on ODT shape, aroma, and taste. Compared with SL and ST, HA endowed ODT with higher fruit aroma, mellow taste, better coordination, and higher sensory evaluation score. Physical–chemical composition analysis showed that SL-fixed orange peel was higher than HA- or ST-fixed peel in the content of polyphenols, flavonoids, soluble protein, hesperidin and limonin, while HA has a higher content of volatile substances and contains more alcohols, aldehydes and ketones, and acid and esters than ST and SL. Activity analysis showed that HA was superior to ST or SL in comprehensive antioxidant activity and inhibitory activity against α-glucosidase. Comprehensive results demonstrated that HA has better performance in improving ODT quality and can replace the traditional SL method in production.

Thiamethoxam, a nicotinic pesticide used worldwide, can cause great harm to the environment and even to human health, and aptamers, known as chemical antibodies, have high affinity and specificity for the target, as well as great potential in detecting small molecules such as pesticides. In this paper, we report a highly sensitive biosensor system for thiamethoxam residue detection based on aptamer technology. After 15 rounds of screening with the pressurized GO-SELEX technology, we found that the aptamer libraries of the 5th and 9th rounds showed high affinity by the capture method. Four candidate aptamers were obtained by high-throughput sequencing and secondary structure prediction. Among them, the aptamer named Thi-5R-18 from the 5th round was demonstrated to possess the highest affinity by isothermal titration calorimetry, with a dissociation constant (Kd) of 4.935 × 10−5 M. The results of molecular docking showed that thiamethoxam and Thi-5R-18 were combined with bases G-15, A-19, and T-71 through hydrogen bonding and π–π interaction.Thi-5R-18 was used as a recognition element to construct a AuNPs colorimetric aptasensor, achieving an ultralow detection limit of 0.37 nM. More importantly, this colorimetric aptasensor can be used for quantitative detection of thiamethoxam on tea leaves, with a recovery of 96.94%~105.86%. This study provides a highly sensitive biosensor for detection of thiamethoxam residue.

The detection of bifenthrin is closely related to the adsorption of SERS substrates. In this study, superparamagnetic Fe3O4@Au MNPs coated with GO were used to detect the adsorption and enrichment of bifenthrin molecules with benzene rings. Firstly, the thermal solvent method synthesized Fe3O4 magnetic nanoparticles (MNPs) with a particle size of ~250 nm. Next, polyethylene imide (PEI) was used as an intermediate layer to modify the surface of Fe3O4 to form a positively charged ultra-thin polymer middle layer. Next, the gold shell was developed by adsorption of ~20 nm AuNPs, without affecting the magnetic properties. Then, the additional amount of colloidal gold and GO on SERS performance was systematically studied. Using crystal violet (CV) as the probe, we investigated the SERS performance of composite nanomaterials. The lowest detected concentration reached 10−8 mol/L, confirming that the composite functional material had good SERS activity and magnetic properties. Finally, the substrate was used to detect bifenthrin in an acetone solution, and the lowest detection concentration was 10−8 mol/L. These results showed that the prepared GO/Fe3O4@Au MNPs were efficient SERS substrates that could detect bifenthrin pesticide residue with high sensitivity.

Tea plants are widely grown all over the world because they are an important economic crop. The purity and authenticity of tea varieties are frequent problems in the conservation and promotion of germplasm resources in recent years, which has brought considerable inconvenience and uncertainty to the selection of parental lines for breeding and the research and cultivation of superior varieties. However, the development of core SNP markers can quickly and accurately identify the germplasm, which plays an important role in germplasm identification and the genetic relationship analysis of tea plants. In this study, based on 179,970 SNP loci from the whole genome of the tea plant, all of 142 cultivars were clearly divided into three groups: Assam type (CSA), Chinese type (CSS), and transitional type. Most CSA cultivars are from Yunnan Province, which confirms that Yunnan Province is the primary center of CSA origin and domestication. Most CSS cultivars are distributed in east China; therefore, we deduced that east China (mainly Zhejiang and Fujian provinces) is most likely the area of origin and domestication of CSS. Moreover, 45 core markers were screened using strict criteria to 179,970 SNP loci, and we analyzed 117 well-Known tea cultivars in China with 45 core SNP markers. The results were as follows: (1) In total, 117 tea cultivars were distinguished by eight markers, which were selected to construct the DNA fingerprint, and the remaining markers were used as standby markers for germplasm identification. (2) Ten pairs of parent and offspring relationships were confirmed or identified, and among them, seven pairs were well-established pedigree relationships; the other three pairs were newly identified. In this study, the east of China (mainly Zhejiang and Fujian provinces) is most likely the area of origin and domestication of CSS. The 45 core SNP markers were developed, which provide a scientific basis at the molecular level to identify the superior tea germplasm, undertake genetic relationship analysis, and benefit subsequent breeding work.

A dsRNA segment was identified in the fungus Melanconiella theae isolated from tea plants. The complete dsRNA sequence, determined by random cloning together with RACE protocol, is 2,461 bp in length with an AU-rich content (62.37%) and comprises a single ORF of 2,265-nucleotides encoding an RNA-dependent RNA-polymerase (RdRp, 754 amino acids in size). The terminus sequences can fold into predicted stable stemloop structures. A BLASTX and phylogenetic analysis revealed the dsRNA genome shows similarities with the RdRp sequences of mitoviruses, with the highest identity of 48% with those of grapevine-associated mitovirus 20 and Colletotrichum fructicola mitovirus 1. Our results reveal a novel member, tentatively named Melanconiella theae mitovirus 1 (MtMV1), belongs to the family Mitoviridae. MtMV1 is capsidless as examined by transmission electron microscope, efficiently transmitted through conidia as 100 conidium-generated colonies were analyzed, and easily eliminated by hyphal tipping method combined with green-leaf tea powder. MtMV1 has a genomic sequence obviously divergent from those of most members in the family Mitoviridae and some unique characteristics unreported in known members. This is the first report of a mycovirus infecting Melanconiella fungi to date. [ABSTRACT FROM AUTHOR]

The oolong tea aroma is shown to consist of cultivar aroma and technical aroma in this study based on the aroma differences between oolong tea products of cultivars of different suitability, as determined by correlation analysis of olfactory, sensory, electronic nose, and GC-MS data. Human senses were significantly affected by the aroma components, which included eight terpene metabolites (β-Ocimene, (Z)-Furan linalool oxide, linalool, (3E)-4,8-Dimethyl-1,3,7-nonatriene, (E)-Pyranoid linalool oxide, γ-Elemene, Humulene, (Z,E)-α-Farnesene), three carotenoid metabolites (β-Ionone, (Z)-Geranylacetone and 6-methyl-5-Hepten -2-one), three lipid metabolites ((Z)-3-Hexenyl (Z)-3-hexenoate, Butanoic acid hexyl ester, and (Z)-Jasmone), four amino acid metabolites (Methyl salicylate, Geranyl isovalerate, indole, and Phenylethyl alcohol), and six thermal reaction products (2-Pentylfuran, Octanal, Decanal, (E,E)-2,4-Nonadienal, (Z)-2-Decenal, and (E)-2-Undecenal). Meanwhile, several aroma compounds (such as (E)-Nerolidol and α-Farnesene), mainly comprising the “technical aroma” formed in the processing mode, were noted to be less closely related to cultivar suitability. This study sheds light on the aroma characteristics of different tea cultivars for oolong tea processing.

Amplicon sequencing is a powerful tool for analyzing the fungal composition inside plants, whereas its application for the identification of etiology for plant diseases remains undetermined. Here, we utilize this strategy to clarify the etiology responsible for tea leaf brown-black spot disease (LBSD), a noticeable disease infecting tea plants etiology that remains controversial. Based on the ITS-based amplicon sequencing analysis, Didymella species were identified as separate from Pestalotiopsis spp. and Cercospora sp., which are concluded as the etiological agents. This was further confirmed by the fungal isolation and their specific pathogenicity on diverse tea varieties. Based on the morphologies and phylogenetic analysis constructed with multi-loci (ITS, LSU, tub2, and rpb2), two novel Didymella species—tentatively named D. theae and D. theifolia as reference to their host plants—were proposed and characterized. Here, we present an integrated approach of ITS-based amplicon sequencing in combination with fungal isolation and fulfillment of Koch’s postulates for etiological identification of tea plant disease, revealing new etiology for LBSD. This contributes useful information for further etiological identification of plant disease based on amplicon sequencing, as well as understanding, prevention, and management of this economically important disease.

Changes in key odorants of shaken black tea (SBT) during its manufacture were determined using headspace solid-phase microextraction (HS-SPME) combined with comprehensive two-dimensional gas chromatography–time-of-flight mass spectrometry (GC × GC–TOFMS) and multivariate data analysis. A total of 241 volatiles was identified, comprising 49 aldehydes, 40 esters, 29 alcohols, 34 ketones, 30 aromatics, 24 alkenes, 17 alkanes, 13 furans, and 5 other compounds. A total of 27 volatiles had average relative odor activity values (rOAVs) greater than 1, among which (E)-β-ionone, (E,Z)-2,6-nonadienal, and 1-octen-3-one exhibited the highest values. According to the criteria of variable importance in projection (VIP) > 1, p < 0.05, and |log2FC| > 1, 61 discriminatory volatile compounds were screened out, of which 26 substances were shared in the shaking stage (FL vs. S1, S1 vs. S2, S2 vs. S3). The results of the orthogonal partial least squares discriminate analysis (OPLS-DA) differentiated the influence of shaking, fermentation, and drying processes on the formation of volatile compounds in SBT. In particular, (Z)-3-hexenol, (Z)-hexanoic acid, 3-hexenyl ester, (E)-β-farnesene, and indole mainly formed in the shaking stage, which promoted the formation of the floral and fruity flavor of black tea. This study enriches the basic theory of black tea flavor quality and provide the theoretical basis for the further development of aroma quality control.