The effect of maturity of tea leaves and processing methods on the formation of milky flavor in white tea - A metabolomic study.

Targeted metabolomics combined with chemometrics were applied to investigate the flavor profiles of 4 white tea samples, which were produced from different maturity fresh tea leaves with different withering methods. Mature leaves that underwent novel withering process at higher temperature (28-30℃) and humidity (75 ± 3 %) (MN) were characterized by intense milky flavor. The content of free amino acids, catechins, and soluble sugars in MN were significantly lower than that in the other 3 tea samples, resulting in a sweet and mellow taste with low bitterness. Meanwhile, MN possessed the highest intensity of milky aroma, which could be mainly attributed to the existence of dihydro-5-pentyl-2(3H)-furanone and 2-pentyl-furan as the key volatile substances with coconut and creamy fragrance. These findings provide insight into the substance foundations of milky flavor, and identified leaf maturity and processing method as the determining factors of the milk-flavored white tea (MFWT).

Study on the dynamic change of volatile components of white tea in the pile-up processing based on sensory evaluation and ATD-GC-MS Technology.

The pile-up processing has a great impact on the flavor of white tea. To investigate the effects of the volatile accumulation of white tea with different piling thickness treatments, tea leaves from different thickness treatments were subjected to sensory quantitative description analysis and ATD-GC-MS detection in this study. As a result, 122 volatile components were identified from white tea with different treatments. A total of 8 key compounds, including isovaleraldehyde, isobutyraldehyde, 2-methyl-butanal, 1-octene-3-ol, linalool, pentanoic acid, hexanal and 1-hexanol were screened out using multivariate statistical analysis, which were characteristic components of grassy, floral-fruity, pekoe aroma and sweet flavors. The results of the selected key characteristic volatile compounds were consistent with the sensory quantitative description. The aroma of mid-pile dried tea (MD) was exhibited a harmonious and pleasant overall flavor. This study provides a novel insight into the accumulation of volatile during the withering step of white tea production.

Comparative volatiles profiling in milk-flavored white tea and traditional white tea Shoumei via HS-SPME-GC-TOFMS and OAV analyses.

White tea is a mildly fermented tea processed with withering and drying. Milk-flavored white tea has a unique milk flavor compared to the traditional white tea. Little is known about the aromas that make white tea taste milky. Here we conducted the volatile profiling via headspace solid-phase microextraction (HS-SPME)-gas chromatography-time-of-flight mass spectrometry (GC-TOFMS) and chemometrics to explore the key volatiles making milk-flavored white tea taste milky. Sixty-seven volatiles were identified, with 7 volatiles (OAV > 1 and VIP > 1) were characterized as the typical aromas. Green and light fruity scent volatiles, such as methyl salicylate, benzyl alcohol, and phenylethyl alcohol, were richer in TFs than MFs. Strong fruity and cheese aromas, such as dihydro-5-pentyl-2(3H)-furanone, 2-pentyl-furan, (E)-6,10-dimethyl-5,9-undecadien-2-one, and hexanal, were more abundant in MFs than TFs. Dihydro-5-pentyl-2(3H)-furanone, recognized as coconut and creamy aroma, should be the essential volatile for milky flavor. Also, (E)-6,10-dimethyl-5,9-undecadien-2-one and 2-pentyl-furan may contribute to the milk scent formation.

Dynamic change of the carotenoid metabolic pathway profile during oolong tea processing with supplementary LED light.

Carotenoid-derived volatiles are important contributors to tea aroma quality. However, the profile of the carotenoid pathway and carotenoid-derived volatiles (CDVs) artificial regulation in oolong tea processing has yet to be investigated. In the present work, the content and varieties of carotenoid-derived volatiles, the genome-wide identification of carotenoid cleavage dioxygenase (CsCCD) gene family, the expression level of CsCCD and other key genes in the carotenoid pathway, and the profile of carotenoid substances were analyzed by multi-omics and bioinformatics methods with innovative postharvest supplementary LED light during oolong tea processing. The results showed that during oolong tea processing, a total of 17 CDVs were identified. The content of ß-ionone increased up to 26.07 times that of fresh leaves and its formation was significantly promoted with supplementary LED light from 0.54 µg/g to 0.83 µg/g in the third turning over treatment. A total of 11 CsCCD gene family members were identified and 119 light response cis-acting regulatory elements of CsCCD were found. However, the expression level of most genes in the carotenoid pathway including CsCCD were reduced due to mechanical stress. 'Huangdan' fresh tea leaves had a total of 1 430.46 µg/g 22 varieties of carotenoids, which mainly composed of lutein(78.10%), ß-carotene(8.24%) and zeaxanthin(8.18%). With supplementary LED light, the content of antherxanthin and zeaxanthin in xanthophyll cycle was regulated and CDVs such as α-ionone, ß-ionone, pseudoionone, damascenone, 6,10-dimethyl-5,9-undecadien-2-one, citral, geranyl acetate and α-farnesene were promoted significantly in different phases during oolong tea processing. Our results revealed the profile of the carotenoid metabolism pathway in oolong tea processing from the perspective of precursors, gene expression and products, and put forward an innovative way to improve CDVs by postharvest supplementary LED light.

Metabolomics integrated with machine learning to discriminate the geographic origin of Rougui Wuyi rock tea.

The geographic origin of agri-food products contributes greatly to their quality and market value. Here, we developed a robust method combining metabolomics and machine learning (ML) to authenticate the geographic origin of Wuyi rock tea, a premium oolong tea. The volatiles of 333 tea samples (174 from the core region and 159 from the non-core region) were profiled using gas chromatography time-of-flight mass spectrometry and a series of ML algorithms were tested. Wuyi rock tea from the two regions featured distinct aroma profiles. Multilayer Perceptron achieved the best performance with an average accuracy of 92.7% on the training data using 176 volatile features. The model was benchmarked with two independent test sets, showing over 90% accuracy. Gradient Boosting algorithm yielded the best accuracy (89.6%) when using only 30 volatile features. The proposed methodology holds great promise for its broader applications in identifying the geographic origins of other valuable agri-food products.

Aroma characteristics of Wuyi rock tea prepared from 16 different tea plant varieties.

Wuyi rock tea (WRT) is famous for its long history and unique characteristic of floral, fruity and nutty flavors. This study investigated the aroma characteristics of WRTs prepared from 16 different oolong tea plant varieties. The sensory evaluation results showed that all WRTs had an 'Yan flavor' taste, and the odor was strong and lasting. Roasted, floral and fruity odors were the prime aroma profiles for WRTs. Furthermore, a total of 368 volatile compounds were detected using HS-SPME-GC-MS and analyzed with OPLS-DA and HCA methods. The volatile compounds heterocyclic compounds, esters, hydrocarbons, terpenoids and ketones were the major aromatic components of the WRTs. Specifically, the volatile profiles among newly selected cultivars were comparatively analyzed, and 205 differential volatile compounds were found with variable importance in the projection (VIP) values above 1.0. These results indicated that the aroma profiles of WRTs were mainly dependent on the cultivar specificities of volatile compounds.

Integrated metabolomic and transcriptomic analysis reveal the effect of mechanical stress on sugar metabolism in tea leaves (Camellia sinensis) post-harvest.

Sugar metabolites not only act as the key compounds in tea plant response to stress but are also critical for tea quality formation during the post-harvest processing of tea leaves. However, the mechanisms by which sugar metabolites in post-harvest tea leaves respond to mechanical stress are unclear. In this study, we aimed to investigate the effects of mechanical stress on saccharide metabolites and related post-harvest tea genes. Withered (C15) and mechanically-stressed (V15) for 15 min Oolong tea leaves were used for metabolome and transcriptome sequencing analyses. We identified a total of 19 sugar metabolites, most of which increased in C15 and V15. A total of 69 genes related to sugar metabolism were identified using transcriptome analysis, most of which were down-regulated in C15 and V15. To further understand the relationship between the down-regulated genes and sugar metabolites, we analyzed the sucrose and starch, galactose, and glycolysis metabolic pathways, and found that several key genes of invertase (INV), α-amylase (AMY), ß-amylase (BMY), aldose 1-epimerase (AEP), and α-galactosidase (AGAL) were down-regulated. This inhibited the hydrolysis of sugars and might have contributed to the enrichment of galactose and D-mannose in V15. Additionally, galactinol synthase (Gols), raffinose synthase (RS), hexokinase (HXK), 6-phosphofructokinase 1 (PFK-1), and pyruvate kinase (PK) genes were significantly upregulated in V15, promoting the accumulation of D-fructose-6-phosphate (D-Fru-6P), D-glucose-6-phosphate (D-glu-6P), and D-glucose. Transcriptome and metabolome association analysis showed that the glycolysis pathway was enhanced and the hydrolysis rate of sugars related to hemicellulose synthesis slowed in response to mechanical stress. In this study, we explored the role of sugar in the response of post-harvest tea leaves to mechanical stress by analyzing differences in the expression of sugar metabolites and related genes. Our results improve the understanding of post-harvest tea's resistance to mechanical stress and the associated mechanism of sugar metabolism. The resulting treatment may be used to control the quality of Oolong tea.

Secondary injury and pro-inflammatory macrophages increase osteophyte growth and fracture healing in canine atrophic nonunion.

OBJECTIVES: In this study, we used a canine high-energy fracture model to examine the relationship between the early inflammatory reaction in adjacent tissues and the ability for osteophyte growth, aiming to identify causes that lead to atrophic nonunion inflammatory disease and to provide new strategies for prevention and treatment. MATERIALS AND METHODS: Forty-eight models of canine femoral high energy fractures were prepared and randomly divided into groups A and B (n=24 in each group). Dogs in both groups underwent open reduction and 6-hole plate internal fixation. Group A models were re-opened, and muscle near the bone was scraped at 14 d after the operation. On days 3, 17, 28, and 42 after fracture, 6 experimental dogs were euthanized per group, and the fracture specimens were used to examine pathologic changes and the growth of callus in the fractured end and its adjacent tissues. RESULTS: At day 14, neutrophil infiltration, with no macrophage recruitment, no mesenchymal cell proliferation, and no fracture healing cascade were observed in the adjacent tissues of both groups. Immediately after the second injury was performed in group A, many macrophages were seen, and mesenchymal cells proliferated, which initiated vigorous osteophyte growth and led to osteophyte healing. Atrophic nonunion was observed in group B without secondary injury. CONCLUSION: Macrophage recruitment deficiency in adjacent soft tissue in early surgery for high-energy fractures may be an important cause of atrophic nonunion. Secondary injury inflammation can effectively recruit mononuclear macrophages, generate osteoclasts, re-initiate the growth of osteophytes, and promote fracture healing.

Transcriptome and metabolite analyses provide insights into zigzag-shaped stem formation in tea plants (Camellia sinensis).

BACKGROUND: Shoot orientation is important for plant architecture formation, and zigzag-shaped shoots are a special trait found in many plants. Zigzag-shaped shoots have been selected and thoroughly studied in Arabidopsis; however, the regulatory mechanism underlying zigzag-shaped shoot development in other plants, especially woody plants, is largely unknown. RESULTS: In this study, tea plants with zigzag-shaped shoots, namely, Qiqu (QQ) and Lianyuanqiqu (LYQQ), were investigated and compared with the erect-shoot tea plant Meizhan (MZ) in an attempt to reveal the regulation of zigzag-shaped shoot formation. Tissue section observation showed that the cell arrangement and shape of zigzag-shaped stems were aberrant compared with those of normal shoots. Moreover, a total of 2175 differentially expressed genes (DEGs) were identified from the zigzag-shaped shoots of the tea plants QQ and LYQQ compared to the shoots of MZ using transcriptome sequencing, and the DEGs involved in the "Plant-pathogen interaction", "Phenylpropanoid biosynthesis", "Flavonoid biosynthesis" and "Linoleic acid metabolism" pathways were significantly enriched. Additionally, the DEGs associated with cell expansion, vesicular trafficking, phytohormones, and transcription factors were identified and analysed. Metabolomic analysis showed that 13 metabolites overlapped and were significantly changed in the shoots of QQ and LYQQ compared to MZ. CONCLUSIONS: Our results suggest that zigzag-shaped shoot formation might be associated with the gravitropism response and polar auxin transport in tea plants. This study provides a valuable foundation for further understanding the regulation of plant architecture formation and for the cultivation and application of horticultural plants in the future.

Integrative Transcriptomic and Metabolic Analyses Provide Insights into the Role of Trichomes in Tea Plant (Camellia Sinensis).

Trichomes, which develop from epidermal cells, are regarded as one of the key features that are involved in the evaluation of tea quality and tea germplasm resources. The metabolites from trichomes have been well characterized in tea products. However, little is known regarding the metabolites in fresh tea trichomes and the molecular differences in trichomes and tea leaves per se. In this study, we developed a method to collect trichomes from tea plant tender shoots, and their main secondary metabolites, including catechins, caffeine, amino acids, and aroma compounds, were determined. We found that the majority of these compounds were significantly less abundant in trichomes than in tea leaves. RNA-Seq was used to investigate the differences in the molecular regulatory mechanism between trichomes and leaves to gain further insight into the differences in trichomes and tea leaves. In total, 52.96 Gb of clean data were generated, and 6560 differentially expressed genes (DEGs), including 4471 upregulated and 2089 downregulated genes, were identified in the trichomes vs. leaves comparison. Notably, the structural genes of the major metabolite biosynthesis pathways, transcription factors, and other key DEGs were identified and comparatively analyzed between trichomes and leaves, while trichome-specific genes were also identified. Our results provide new insights into the differences between tea trichomes and leaves at the metabolic and transcriptomic levels, and open up new doors to further recognize and re-evaluate the role of trichomes in tea quality formation and tea plant growth and development.

Neuroprotective and Anti-Amyloid ß Effect and Main Chemical Profiles of White Tea: Comparison Against Green, Oolong and Black Tea.

White tea (WT) is one of six tea types originally derived from Fujian Province, China. White tea is known for its health-promoting properties. However, the neuroprotective and anti-aggregatory properties of WT against the hallmark toxic Alzheimer's protein, Aß have not been investigated. In this study, WT, green tea (GT), oolong tea (OT) and black tea (BT) were manufactured using tea leaves from the cultivar Camellia sinensis (Jin Guanyin). The protective effects of these tea extracts were then studied under oxidative stress conditions via t-bhp and H2O2 exposure, in addition to Aß treatment using a PC-12 cell model. Each tea type failed to rescue PC-12 cells from either t-bhp or H2O2-mediated toxicity, however each extract exerted significant protection against Aß-evoked neurotoxicity. Results of the Thioflavin T Kinetic (ThT) and TEM assay showed that Aß aggregate formation was inhibited by each tea type. Additionally, TEM also supported the different anti-aggregatory effect of WT by modifying Aß into an amorphous and punctate aggregate morphology. Higher accumulated precedent or potential neuroprotective compounds in WT, including ECG''3Me, 8-C-ascorbyl-EGCG, GABA and Gln, in addition to flavonol or flavone glycosides detected by using UPLC-QTOF-MS and UPLC-QqQ-MS, may contribute to a favourable anti-aggregative and neuroprotective effect of WT against Aß.

Expression patterns of alpha-amylase and beta-amylase genes provide insights into the molecular mechanisms underlying the responses of tea plants (Camellia sinensis) to stress and postharvest processing treatments.

MAIN CONCLUSION: The alpha-amylase and beta-amylase genes have been identified from tea plants, and their bioinformatic characteristics and expression patterns provide a foundation for further studies to elucidate their biological functions. Alpha-amylase (AMY)- and beta-amylase (BAM)-mediated starch degradation plays central roles in carbohydrate metabolism and participates extensively in the regulation of a wide range of biological processes, including growth, development and stress response. However, the AMY and BAM genes in tea plants (Camellia sinensis) are poorly understood, and the biological functions of these genes remain to be elucidated. In this study, three CsAMY and nine CsBAM genes from tea plants were identified based on genomic and transcriptomic database analyses, and the genes were subjected to comprehensive bioinformatic characterization. Phylogenetic analysis showed that the CsAMY proteins could be clustered into three different subfamilies, and nine CsBAM proteins could be classified into four groups. Putative catalytically active proteins were identified based on multiple sequence alignments, and the tertiary structures of these proteins were analyzed. Cis-element analysis indicated that CsAMY and CsBAM were extensively involved in tea plant growth, development and stress response. In addition, the CsAMY and CsBAM genes were differentially expressed in various tissues and were regulated by stress treatments (e.g., ABA, cold, drought and salt stress), and the expression patterns of these genes were associated with the postharvest withering and rotation processes. Taken together, our results will enhance the understanding of the roles of the CsAMY and CsBAM gene families in the growth, development and stress response of tea plants and of the potential functions of these genes in determining tea quality during the postharvest processing of tea leaves.

An Efficient Extraction Method for Fragrant Volatiles from Jasminum sambac (L.) Ait.

The sweet smell of aroma of Jasminum sambac (L.) Ait. is releasing while the flowers are blooming. Although components of volatile oil have been extensively studied, there are problematic issues, such as low efficiency of yield, flavour distortion. Here, the subcritical fluid extraction (SFE) was performed to extract fragrant volatiles from activated carbon that had absorbed the aroma of jasmine flowers. This novel method could effectively obtain main aromatic compounds with quality significantly better than solvent extraction (SE). Based on the analysis data with response surface methodology (RSM), we optimized the extraction conditions which consisted of a temperature of 44°C, a solvent-to-material ratio of 3.5:1, and an extraction time of 53 min. Under these conditions, the extraction yield was 4.91%. Furthermore, the key jasmine essence oil components, benzyl acetate and linalool, increase 7 fold and 2 fold respectively which lead to strong typical smell of the jasmine oil. The new method can reduce spicy components which lead to the essential oils smelling sweeter. Thus, the quality of the jasmine essence oil was dramatically improved and yields based on the key component increased dramatically. Our results provide a new effective technique for extracting fragrant volatiles from jasmine flowers.