Daxiang Li

New triterpenoids and other constituents from a special microbial-fermented tea-Fuzhuan brick tea.

Fuzhuan brick tea, a special microbial-fermented tea prepared from the leaves of Camellia sinensis var. sinensis, is a traditional beverage used in China throughout history. Phytochemical investigation of this material led to the identification of three new triterpenoids, 3beta,6alpha,13beta-trihydroxyolean-7-one (1), 3beta-acetoxy-6alpha,13beta-dihydroxyolean-7-one (2), and 3beta-O-(8-hydroxyoctanoyl)-12-oleanene (3), together with 11 known compounds, friedelin (4), beta-amyrone (5), beta-amyrin (6), alpha-spinasterone (7), alpha-spinasterol (8), 22,23-dihydro-alpha-spinasterone (9), 22,23-dihydro-alpha-spinasterol (10), alpha-phytol (11), alpha-tocopherol (12), alpha-tocoquinone (13), and caffeine (14). The structures of 1-13 were determined by spectroscopic and chemical methods. Compounds 1 and 2 are the first two examples of triterpenoids possessing a 6-hydroxy-7-one function. All of the compounds, except 6, 8, 10, 11, and 14, were isolated from tea and Camellia spp. for the first time. The antibacterial activities of 1 were assessed against some enteric pathogenic microbes. Compound 1 showed no cytotoxic activity against A-549, Bel-7402, and HCT-8 cell lines.

Discrimination of the production season of Chinese green tea by chemical analysis in combination with supervised pattern recognition.

High-performance liquid chromatography (HPLC) has been used to quantify levels of free amino acids, catechins, and caffeine in Chinese green tea. Levels of free amino acids and catechins in green tea leaves show obvious variation from spring to summer, which is useful information to identify the production season of commercial green tea. Supervised pattern recognition methods such as the K-nearest neighbor (KNN) method and Bayesian discriminant method (a type of linear discriminant analysis (LDA)) were used to discriminate between the production seasons of Chinese green tea. The optimal accuracy of the KNN method was ≤97.61 and ≤94.80% as validated by resubstitution and cross-validation tests, respectively, and that of LDA was ≤95.22 and ≤93.54%, respectively. Compared with LDA, the KNN method did not require a Gaussian distribution and was more accurate than LDA. The KNN method in combination with chemical analysis is recommended for discrimination of the production seasons of Chinese green tea.

Adsorptive removal of fluoride from drinking water using porous starch loaded with common metal ions

In this study, porous corn starch was loaded with Zr, Al, Fe or La to produce the composites PS-Zr, PS-Al, PS-Fe and PS-La. Fluoride adsorption from water was tested at different biosorbent dosages, contact times, solution pH values and initial fluoride concentrations. The biosorbents were characterized by microscopy and spectroscopy. PS-Zr was shown superior defluoridation capacity over a pH range of 3.0-9.0. The adsorption process could be described by the Langmuir isotherm model and the Lagergren pseudo-second-order kinetic model. The maximum fluoride adsorption capacity calculated to be for PS-Zr was 25.41mg/g. Our results revealed that PS-Zr could be employed as an effective biosorbent for removal of fluoride from drinking water.

Draft genome sequence of Camellia sinensis var. sinensis provides insights into the evolution of the tea genome and tea quality

Significance A high-quality genome assembly of Camellia sinensis var. sinensis facilitates genomic, transcriptomic, and metabolomic analyses of the quality traits that make tea one of the world’s most-consumed beverages. The specific gene family members critical for biosynthesis of key tea metabolites, monomeric galloylated catechins and theanine, are indicated and found to have evolved specifically for these functions in the tea plant lineage. Two whole-genome duplications, critical to gene family evolution for these two metabolites, are identified and dated, but are shown to account for less amplification than subsequent paralogous duplications. These studies lay the foundation for future research to understand and utilize the genes that determine tea quality and its diversity within tea germplasm. Tea, one of the world’s most important beverage crops, provides numerous secondary metabolites that account for its rich taste and health benefits. Here we present a high-quality sequence of the genome of tea, Camellia sinensis var. sinensis (CSS), using both Illumina and PacBio sequencing technologies. At least 64% of the 3.1-Gb genome assembly consists of repetitive sequences, and the rest yields 33,932 high-confidence predictions of encoded proteins. Divergence between two major lineages, CSS and Camellia sinensis var. assamica (CSA), is calculated to ∼0.38 to 1.54 million years ago (Mya). Analysis of genic collinearity reveals that the tea genome is the product of two rounds of whole-genome duplications (WGDs) that occurred ∼30 to 40 and ∼90 to 100 Mya. We provide evidence that these WGD events, and subsequent paralogous duplications, had major impacts on the copy numbers of secondary metabolite genes, particularly genes critical to producing three key quality compounds: catechins, theanine, and caffeine. Analyses of transcriptome and phytochemistry data show that amplification and transcriptional divergence of genes encoding a large acyltransferase family and leucoanthocyanidin reductases are associated with the characteristic young leaf accumulation of monomeric galloylated catechins in tea, while functional divergence of a single member of the glutamine synthetase gene family yielded theanine synthetase. This genome sequence will facilitate understanding of tea genome evolution and tea metabolite pathways, and will promote germplasm utilization for breeding improved tea varieties.

Molecular Cloning and Characterization of Galactinol Synthases in Camellia sinensis with Different Responses to Biotic and Abiotic Stressors

Galactinol synthase (GolS) is a key biocatalyst for the synthesis of raffinose family oligosaccharides (RFOs). RFOs accumulation plays a critical role in abiotic stress adaptation, but the relationship between expression of GolS genes and biotic stress adaptation remains unclear. In this study, two CsGolS genes were found to be highly up-regulated in a transcriptome library of Ectropic oblique-attacked Camellia sinensis. Three complete CsGolS genes were then cloned and characterized. Gene transcriptional analyses under biotic and abiotic stress conditions indicated that the CsGolS1 gene was sensitive to water deficit, low temperature, and abscisic acid, while CsGolS2 and CsGolS3 genes were sensitive to pest attack and phytohormones. The gene regulation and RFOs determination results indicated that CsGolS1 was primarily related to abiotic stress and CsGolS2 and CsGolS3 were related to biotic stress. GolS-mediated biotic stress adaptations have not been studied in depth, so further analysis of this new biological function is required.

Effect of Medium Supplements on Agrobacterium rhizogenes Mediated Hairy Root Induction from the Callus Tissues of Camellia sinensis var. sinensis.

Tea (Camellia sinensis L.) is recalcitrant to Agrobacterium-mediated genetic transformation largely due to the bactericidal effects of tea polyphenols and phenolics oxidation induced by necrosis of explant tissue over the process of transformation. In this study, different antioxidants/adsorbents were added as supplements to the co-cultivation and post co-cultivation media to overcome these problems for the transformation improvement. Tea-cotyledon-derived calli were used as explants and Agrobacterium rhizognes strain ATCC 15834 was used as a mediator. Results showed that Agrobacterium growth, virulence (vir) gene expression and browning of explant tissue were greatly influenced by different supplements. Murashige and Skoog (MS) basal salts medium supplemented with 30 g·L−1 sucrose, 0.1 g·L−1 l-glutamine and 5 g·L−1 polyvinylpolypyrrolidone (PVPP) as co-cultivation and post co-cultivation media could maintain these parameters better that ultimately led to significant improvement of hairy root generation efficiency compared to that in the control (MS + 30 g·L−1 sucrose). Additionally, the reporter genes β-glucuronidase (gusA) and cyan fluorescent protein (cfp) were also stably expressed in the transgenic hairy roots. Our study would be helpful in establishing a feasible approach for tea biological studies and genetic improvement of tea varieties.

Brewing and volatiles analysis of three tea beers indicate a potential interaction between tea components and lager yeast

Green tea, oolong tea and black tea were separately introduced to brew three kinds of tea beers. A model was designed to investigate the tea beer flavour character. Comparison of the volatiles between the sample of tea beer plus water mixture (TBW) and the sample of combination of tea infusion and normal beer (CTB) was accomplished by triangular sensory test and HS-SPME GC-MS analysis. The PCA of GC-MS data not only showed a significant difference between volatile features of each TBW and CTB group, but also suggested some key compounds to distinguish TBW from CTB. The results of GC-MS showed that the relative concentrations of many typical tea volatiles were significantly changed after the brewing process. More interestingly, the behaviour of yeast fermentation was influenced by tea components. A potential interaction between tea components and lager yeast could be suggested.

Critical factors determining fluoride concentration in tea leaves produced from Anhui province, China.

This study investigated the fluoride present in tea plants (Camellia sinensis (L.) O. Kuntze) and its relationship to soils, varieties, seasons and tea leaf maturity. The study also explored how different manufacturing processes affect the leaching of fluoride into tea beverages. The fluoride concentration in the tea leaves was significantly correlate to the concentration of water-soluble fluoride in the soil. Different tea varieties accumulated different levels of fluoride, with varieties, Anji baicha having the highest and Nongkang zao having the lowest fluoride concentration. In eight different varieties of tea plant harvested over three tea seasons, fluoride concentration were highest in the summer and lowest in the spring in china. The fluoride concentration in tea leaves was directly related to the maturity of the tea leaves at harvest. Importantly, the tea manufacturing process did not introduced fluoride contamination. The leaching of fluoride was 6.8% and 14.1% higher in black and white tea, respectively, than in fresh tea leaves. The manufacturing step most affecting the leaching of fluoride into tea beverage was withering used in white, black and oolong tea rather than rolling or fermentation. The exposure and associated health risks for fluoride concentration in infusions of 115 commercially available teas from Chinese tea markets was determined. The fluoride concentration ranged from 5.0 to 306.0mgkg(-1), with an average of 81.7mgkg(-1). The hazard quotient (HQ) of these teas indicated that there was no risk of fluorosis from drinking tea, based on statistical analysis by Monte Carlo simulation.

Dietary Supplement of Large Yellow Tea Ameliorates Metabolic Syndrome and Attenuates Hepatic Steatosis in db/db Mice

Yellow tea has been widely recognized for its health benefits. However, its effects and mechanism are largely unknown. The current study investigated the mechanism of dietary supplements of large yellow tea and its effects on metabolic syndrome and the hepatic steatosis in male db/db mice. Our data showed that dietary supplements of large yellow tea and water extract significantly reduced water intake and food consumption, lowered the serum total and low-density lipoprotein cholesterol and triglyceride levels, and significantly reduced blood glucose level and increased glucose tolerance in db/db mice when compared to untreated db/db mice. In addition, the dietary supplement of large yellow tea prevented the fatty liver formation and restored the normal hepatic structure of db/db mice. Furthermore, the dietary supplement of large yellow tea obviously reduced the lipid synthesis related to gene fatty acid synthase, the sterol regulatory element-binding transcription factor 1 and acetyl-CoA carboxylase α, as well as fatty acid synthase and sterol response element-binding protein 1 expression, while the lipid catabolic genes were not altered in the liver of db/db mice. This study substantiated that the dietary supplement of large yellow tea has potential as a food additive for ameliorating type 2 diabetes-associated symptoms.

Large Yellow Tea Attenuates Macrophage-Related Chronic Inflammation and Metabolic Syndrome in High-Fat Diet Treated Mice

Large yellow tea is a traditional beverage in China with a unique toasty flavor. A preliminary study using 3T3-L1 cells indicated that large yellow tea possessed more potent lipid-lowering efficacy than green, black, dark, and white teas. In the present study we further investigated its influence on metabolic syndrome in a high-fat diet (HFD) mouse model with an emphasis on dose response. Thirty-two C57BL/6 male mice were randomly divided into 4 groups: low-fat diet (LFD), HFD, HFD + 2.5% large yellow tea hot-water extract (YT, equivalent to 10 cups of tea daily for humans), HFD + 0.5% YT. Our data indicated that YT treatment for 12 weeks significantly reduced body weight, liver weight, and adipose tissue weight of the mice; lowered serum insulin and leptin; and raised serum adiponectin with dose effect. H&E staining showed that the HFD group exhibited significant enlargement of adipose cell sizes and the corresponding decrease of adipose cell numbers, which were dose-dependently attenuated in both YT groups. IHC results revealed that YT decreased macrophage recruitment in the liver, epididymal adipose tissue, and subcutaneous adipose tissue and depressed serum inflammatory cytokines including TNF-α, MCP-1, IFN-γ, IL-6, and IL-1β, in a dose-dependent manner. In addition, YT decreased serum glucose, TC, TG, LDL-C, and HDL-C, as well as ameliorated glucose intolerance and insulin resistance independent of dose. Overall, YT would be a unique tea with dose-independent antihyperglycemic and robust lipid-lowering efficacies.