Genetic analyses of ancient tea trees provide insights into the breeding history and dissemination of Chinese Assam tea(Camellia sinensis var.assamica)

Chinese Assam tea(Camellia sinensis var.assamica)is an important tea crop with a long history of cultivation in Yunnan,China.Despite its potential value as a genetic resource,its genetic diversity and domestication/breeding history remain unclear.To address this issue,we genotyped 469 ancient tea plant trees representing 26 C.sinensis var.assamica populations,plus two of its wild relatives(six and three populations of C.taliensis and C.crassicolumna,respectively)using 16 nuclear microsatellite loci.Results showed that Chinese Assam tea has a relatively high,but comparatively lower gene diversity(H_(S)=0.638)than the wild relative C.crassicolumna(H_S=0.658).Clustering in STRUCTURE indicated that Chinese Assam tea and its two wild relatives formed distinct genetic groups,with considerable interspecific introgression.The Chinese Assam tea accessions clustered into three gene pools,corresponding well with their geographic distribution.However,New Hybrids analysis indicated that 68.48%of ancient Chinese Assam tea plants from Xishuangbanna were genetic intermediates between the Puer and Lincang gene pools.In addition,10%of the ancient Chinese Assam tea individuals were found to be hybrids between Chinese Assam tea and C.taliensis.Our results suggest that Chinese Assam tea was domesticated separately in three gene pools(Puer,Lincang and Xishuangbanna)in the Mekong River valley and that the hybrids were subsequently selected during the domestication process.Although the domestication history of Chinese Assam tea in southwestern Yunnan remains complex,our results will help to identify valuable genetic resources that may be useful in future tea breeding programs.

The protective roles of tea tree oil extracts in bovine mammary epithelial cells and polymorphonuclear leukocytes

Background: Tea tree oil(TTO) plays an important role in antibacterial activity and alleviating the inflammatory responses. Bovine mammary epithelium and polymorphonuclear leukocytes(PMNL) can actively respond to bovine mastitis infection. However, regulatory effects of TTO extracts on the innate immune response of bovine mammary epithelial cells(BMECs) and PMNL remain not reported. Therefore, aim of the study was to evaluate the effects of TTO extracts on the m RNA levels of the genes involved in the innate immune response of BMECs and PMNL.Results: Our results demonstrated that addition of 0.025% and 0.05% TTO increased the proliferation of BMECs, and significantly enhanced(P < 0.05) the viability of BMECs exposed to Staphylococcus aureus(S. aureus). An inhibitory effect was observed against the growth of S. aureus by TTO incubation. The 0.05% TTO reduced S. aureus biofilm formation, association and invasion of S. aureus to BMECs, and changed the morphological and structural features of S. aureus. The proinflammatory cytokines IL-1β, IL-6, and TNF-α were decreased(P < 0.001) by the incubation of TTO. Interestingly, the expression of IL-8 known for PMNL chemotactic function was elevated(P < 0.05) by 0.05%TTO treatment. Consistently, 0.05% TTO increased the migration of PMNL in S. aureus-exposed BMECs when compared with S. aureus treatment alone(P < 0.05). In addition, PMNL incubated with 0.05% TTO decreased the levels of NFKB inhibitor alpha(NFKBIA) and TNF-α.Conclusions: Our results indicate that use of TTO can relieve the BMECs pro-inflammatory response caused by S.aureus and promote the migration of PMNL to mount the innate immune responses, and it may be novel strategy for the treatment of bovine mastitis caused by S. aureus.

Determination of the Geographical Origin of Fruits by Using 26S rDNA Fingerprinting of Yeast Communities by PCR-DGGE： An Application to Shea Tree Fruits

International trade intensifies the process of globalization which has extended to the entire planet and extends to the entire planet. In view of the difficulties of installing these documentary systems in developing countries, in particularly the countries of sub-Saharan Africa, the new strategies of traceability emerge. Among the new tools of tracing the products of vegetable origin, a ＂biological code bar＂ based on the analysis of the DNA of micro-organisms present on the fruits is an interesting tool. A molecular technique employing 26S rDNA profiles generated by PCR-DGGE was used to detect the variation in yeast community structures of Shea tree fruits （Vitellaria paradoxa） from Senegal, Ghana and Mali. When the 26S rDNA profiles were analyzed by image analysis and multivariate analysis, distinct microbial communities were detected on Shea tree fruits. The band profiles of yeasts from different countries were specific for each location and could be used as a bar code to discriminate the origin of the fruits. This method is a new traceability tool which provides fruit products with an unique biological bar code and makes it possible to trace back the fruits to their original location.

Estimating the Age of Oil-tea Camellia Trees

To determine the age of oil-tea camellia trees, regression equations including Logistic, Mitscherlich, Gompertz, Korf, and Richards were used to calculate accumulative growth rate using basal trunk disc and investigate the relations between the age of oil-tea camellia trees and their growth rate of secondary trunk. The Gompertz equation Y=71.296 1exp （-3.874 4exp （-0.006 4t）） was the most optimal equation to simulate the accumulative growth rate of basal trunk disc. This equation could be used to estimate the age of oil-tea camellia trees that grow under similar environmental conditions. The Korf equation Y=576.900 1exp （-4.153 0x -0.314 2 ） was the best equation to describe the relation between the age and growth rate of different secondary trunks. With the adjustment coefficient and average growth of different secondary trunk discs, it is possible to predict the age of ancient oil-tea camellia trees that grow under similar environmental conditions. In addition, taking three or more discs from the same diameter group and calculating their average growth rate could lead to more accurate results. For trees that grow in different areas, environmental conditions should be carefully considered when using the above two equations to predict the age of ancient oil-tea camellia trees.

Effects of Intercropping and Shading Systems on Tea Photosynthesis and Respiration

[Objective] The aim was to provide references for constructing compound ecological tea gardens. [Method] In an ecological adult-tea garden, teas shaded by Prunus cerasoides, Prunus L., and Litsea pungens were selected and the teas without shades were taken as a control in order to explore effects of tree shading on photosynthesis, respiration and net photosynthetic intensities. [Result] In a growth cycle of one year, for teas shaded by Prunus cerasoides, Prunus L., and Litsea pungens, respiration intensity was significantly higher than that of the control; net photosynthetic intensity was extremely significant higher; photosynthesis intensity showed none rules. Both of net photosynthetic rate and intensity kept higher in winter of shaded teas. [Conclusion] It is of significance for high-yielding and high-quality teas to reduce respiration consumption and coordinate between photosynthesis and respiration given that tea grows well.

A randomized controlled trial of topical tea tree preparation for MRSA colonized wounds

Background:The prevalence of MRSA(Methicillin-resistant Staphylococcus aureus)colonized wounds in home care residents is expected to grow continuously as a result of the substantial proportion of older people requiring institutionalized care due to chronic disease and declining functional status,which contribute to more frequent skin breakdown and wound formation.Tea tree oil has been claimed to have anti-bacterial,analgesic and anti-inflammatory effects that have been suggested in many in-vitro studies to have good efficacy against MRSA.The aims of this study were to evaluate the effectiveness of 10%topical tea tree preparation to eradicateMRSA and to ascertain its influence on wound healing forMRSA-colonized wounds.Methods:It was a randomized controlled trial,single-blind study.Those with stage II or above MRSA-colonized wounds and who had given their informed consent formed the sample.The determined sample sizewasbasedonthe effect size of our previouspilot study,whichwas 0.46.Five outcome measurements were taken for the MRSA bacterial count and wound healing condition at baseline and at 1-week intervals during the 4-week dressing intervention period.Results:Thirty-two participants were recruited from two non-government nursing homes,16 in the control group and 16 in the tea tree oil group.The control group residents received routine saline gauze dressing,while the tea tree oil group residents received the 10%topical tea tree preparation dressing.In the tea tree oil group,all chronic wounds that had previously been delayed in healing were healed within 28 days without adverse reaction.MRSA was also completely eradicated in 14(87.5%)out of 16 wounds in the group receiving the 10%topical tea tree preparation.

Climatic Zoning of Tea Tree Planting in Hanzhong City

[ Objective] The research aimed to study climatic zoning of tea tree cultivation in Hanzhong. [Method] Based on climate data at 11 meteorological observatories of Hanzhong during 1971 -2010, selecting annual average extreme minimum temperature, annual average temperature, accumulative temperature ≥10 ℃ and annual rainfall as climatic zoning factors, regression model between zoning factors and geographic information was established, and comprehensive climatic zoning indicator of tea tree cultivation in Hanzhong was determined. [ Result] Tea tree cultivation in Hanzhong was divided into suitable, more suitable and unsuitable planting zones by using critical value of climatic zoning. [ Conclusion] The research could provide scientific basis for reasonable planning layout and sustainable development of tea tree in the whole city.

Critical Evaluation of <i>Melaleuca alternifolia</i>: A Review of the Phytochemical Profile, Pharmacological Attributes and Medicinal Properties in the Botanical, Human and Global Perspectives

Melaleuca (tea tree) oil has become increasingly commonly used in recent decades. The essential oil in Australia for the past 120 years is now available globally as an active component in various products. Historically, Melaleuca oil is known for its antiseptic and anti-inflammatory actions. Currently, microwave technology is applied to extract Melaleuca oil, but this extraction technology is not commercially under practice. Traditionally, Melaleuca oil extraction is possible through steam distillation of the Melaleuca alternifolia terminal branches and leaves, and the resultant extract is always either clear or pale yellowish. Melaleuca oil has been promoted as a therapeutic agent because scientific studies indicate that the Rideal-Walker (RW) coefficients of its extract composition include 8 for cymene, 16 for terpineol, 13.5 for tepinen-4-ol and 3.5 for cineole. The mode of action against bacteria is now partially elucidated, and assumptions exist. Hydrocarbons partition into biological membranes to disrupt the vital functions of Melaleuca oil, and also its components behave in the same manner. Therefore, the inhibition of respiration and the leakage of ions or loss of intracellular material and the inability to maintain homeostasis reflect the loss of membrane integrity and lysis in Melaleuca oil products containing lower than usual terpenes concentrations. Melaleuca oil possesses antifungal properties and is known exclusively for the treatment of Candida albicans. This essential oil strongly changes the permeability of Candida albican cells. C. albicans treatment with 0.25% tea tree oil leads to propidium iodide uptake. However, a significant loss of 260-nmlight-absorbing materials after staining with methylene blue occurs after 6 hours. Melaleuca extracts alter the permeability of Candida glabrata that occurs when the membrane is treated with 0.25% Melaleuca oil. Melaleuca oils possess antiviral properties but most findings evidence that this oil fights against both non-enveloped and enveloped viruses, although the range of viruses tested to date is minimal. Melaleuca oil is known for its antiprotozoal activity because it causes a 50% reduction in the growth of protozoa Leishmania major and Trypanosoma brucei at concentrations of 403 mg/ml and 0.5 mg/ml, respectively, based on the studies done (in comparison to controls). An investigation has shown that terpinen-4-ol also contributes significantly to antiprotozoal activity. Tea tree oils at 300 mg/ml killed all cells of Trichomonas vaginalis, and also anecdotal in-vivo studies evidence that Melaleuca oil may be effective in treating infections caused by Trichomonas vaginalis. This review article summarizes the developments in our understanding of the phytochemistry, bioactivity, metabolism and the economic aspects of Melaleuca alternifolia, and it details how Melaleuca alternifolia species have evolved in the ecosystem.

Research on the Preparation of an Anti-Acne Cream and Its Thermal Stability

In this paper, the preparation of tea tree oil and salicylic acid anti-acne cream was studied. The single emulsifier, compound emulsifier, thickener in the acidic system were screened, and the long-term thermal stability was investigated. The experimental results show that when the ether emulsifier, NIKKOL BC-150 and MONTANOV 68, is compounded in a mass ratio of 2:1 and the thickener is SEPIPLUS 400, the obtained product has good thermal stability, and its functional ingredients (tea tree oil and salicylic acid) did not change significantly at high temperatures.

Multiple cells of origin in cholangiocarcinoma underlie biological,epidemiological and clinical heterogeneity

Recent histological and molecular characterization of cholangiocarcinoma(CCA) highlights the heterogeneity of this cancer that may emerge at different sites of the biliary tree and with different macroscopic or morphological features.Furthermore,different stem cell niches have been recently described in the liver and biliarytree,suggesting this as the basis of the heterogeneity of intrahepatic(IH)-and extrahepatic(EH)-CCAs,which are two largely different tumors from both biological and epidemiological points of view.The complexity of the organization of the liver stem cell compartments could underlie the CCA clinical-pathological heterogeneity and the criticisms in classifying primitive liver tumors.These recent advances highlight a possible new classification of CCAs based on cells of origin and this responds to the need of generating homogenous diagnostic,prognostic and,hopefully,therapeutic categories of IH-and EH-CCAs.

Use RAPD Analysis to Classify Tea Trees in Yunnan

RAPD assessment on genetic variations of 45 tea trees in Yunnan was carried out. Eight primers selected from 40 random primers were used to amplify 45 tea samples, and a total of 95 DNA bands were amplified, of which 90(94.7%)were polymorphism. The average number of DNA bands amplified by each primer was 11.5. Based on the results of UPGMA cluster analysis of 95 DNA bands amplified by 8 primers, all the tested materials could be classified into 7 groups including 5 complex groups and 2 simple groups, which was basically identical with morphological classification. In addition, there were some speciations in 2 simple groups.

Variations in seed size and seed mass related to tree growth over 5 years for 23 provenances of Quercus acutissima from across China

The origin of a seed strongly impacts its traits, and both origin and seed traits influence seed germination and seedling development. However, in many instances, this effect on the seedling does not persist into adulthood, and little is known about how seed traits and original environment affect seedling/tree growth over time. In this study, seed size, seed mass, seedling/tree growth and origins were collected and determined for 23 provenances of Quercus acutissima from across China. Origin variables correlated well with seed size and seed mass. In stepwise multiple regressions, a longitudinal aridity index explained 49.2-68.7% of the total variation in seed size and mass, while only seed width was correlated with seedling/tree height (H) and diameter at the ground (D) from seed traits and origins. The total variance in H and D explained by the models decreased over time, for example, the R 2 value of the models for H declined from 0.477 in the first year to 0.224 in the fourth year; no models was significant in the fifth year. These results indicate that seed size, regulated by the longitudinal aridity index strongly impacted seedling and tree growth, but the strength of the influence decreased over time, and disappeared after 4 years.

Cellulase in Anoplophora glabripennis adults fed on original host tree species and non-original host trees

Cellulase activities of Anoplophora glabripennis （Motsch.） adults from two host plants （Populus simonii x p pyramidliscr cv. Opera Hsu. and Salix matsudana Koidz） fed on three different host tree species （Acer negundo Linn., S. matsudana Koidz and P simonii x P pyramidliscr cv. Opera Hsu.） were investigated. Enzyme activities of endoglucanase and β-glucosidase in the intestines of the insects were measured. The results show that there are no statistically significant differences in the enzyme activities of endoglucanase between male and female insects from the two host plants fed on three host trees, neither inβ-glucosidase. No statistically significant differences in the enzyme activities of endoglucanase and β-glucosidase were found between males and females from the two plants fed on three host tree species, except that the endoglucanase activity in males from the opera poplar fed on Hankow willow is lower than that fed on ash-leaf maple and opera poplar.

Feasibility Analysis of in silico Cloning of Functional Candidate Genes in Tea (Camellia sinensis)

[ Objective ] This study aimed to verify the feasibility of in silico cloning of functional candidate genes in tea. [ Method ] Theobroma cacao caffeine syn- thase gene BCS1 was used as a probe to search the established tea EST database using BLAST; 26 tea ESTs highly homologous to BCS1 were obtained, which were assembled using CAP （contig assembly program） of BioEdit software; subsequently, two EST configs harboring ORF were obtained, which were named TCSnewl and TCSnew2, respectively. Nucleotide sequences and deduced amino acid sequences of theses two genes were compared with those of cDNA of tea caffeine synthase gene TCS in the GenBank database that was cloned with experimental biological method. A phylogenetic tree was constructed for homalogous analysis of the deduced amino acid sequences of theses three genes. [ Result] in silico cloning of functional candidate genes in tea using a homologous gene of distantly related species as a probe is a feasible technical means. [ Conclusion] This study provided the basis for in silico cloning of other functional genes in tea.

对92个茶树菇菌株的遗传多样性分析和农艺性状的评价

为了解茶树菇(Agrocybe aegerita)种质资源的遗传多样性和筛选优良的茶树菇新品种,采用菌株拮抗试验方法观察了92株茶树菇菌株间拮抗反应及其类型,ISSR-PCR(inter-simple sequence repeat-PCR)分子标记方法对92株茶树菇菌株的遗传多样性进行了综合分析。拮抗试验将92株茶树菇菌株分为27组;筛选出的20条ISSR引物共扩增出317条清晰条带,多态性条带平均比率为82.60%;在遗传相似系数为0.742时,ISSR分子标记分析可将92株茶树菇划分为6大类群,拮抗试验和ISSR分子标记分析的结果基本一致。通过对比农艺性状分析,初步筛选出滇农5、滇农14、茶5-800、白茶、闽农5及滇农8作为工厂化生产茶树菇菌种。结果表明茶树菇的遗传多样性丰富,结合栽培出菇试验可为茶树菇品种选育和杂交育种的亲本选择提供参考。

不同茶树品种芽型工夫红茶的品质特征

为了解不同茶树品种制作的芽型工夫红茶品质差异,采用高效液相色谱技术、固相微萃取-气相色谱-质谱联用(SPME-GC-MS)技术、定量描述性分析和化学计量学对10个茶树品种芽型工夫红茶的特征滋味物质和特征香气成分进行分析。定量描述性分析结果表明,10个茶树品种芽型工夫红茶的整体香气以甜香、花香为主,滋味以甜醇、浓厚为主。不同样品中检测的16种内含成分(非挥发性成分)存在一定的差异,平均变异系数为21.63%,其中茶黄素的差异最大;在10个茶树品种红茶中共检测出49种挥发性物质,其中质量分数最高的挥发性物质类型为醇类化合物,其次为酯类化合物,酮类化合物质量分数最低,其中香叶醇质量分数最高;正交偏最小二乘判别分析(orthogonal partial least squares-discriminant analysis,OPLS-DA)能将‘金牡丹‘’福云7号’与其他茶树品种红茶区分开,且筛选出12种与芽型工夫红茶茶树品种密切相关的标志差异物。相关性分析结果表明,咖啡碱和茶多酚与回甘呈显著正相关,咖啡碱、可可碱与醇度呈显著正相关,香叶醇、顺-α,α-5-三甲基-5-乙烯基四氢化呋喃-2-甲醇、水杨酸甲酯、芳樟醇与花果香呈正相关。该研究为芽型工夫红茶的生产加工和品质提升提供了理论依据。

万州不同种植区域川中小叶种红茶滋味品质分析

为探明万州川中小叶种制作红茶滋味品质,该文以9个不同区域和表型特征的川中小叶种制备红茶,采用靶向检测方法,分析茶汤中儿茶素类、游离氨基酸类、生物碱类、黄酮醇及其苷类、有机酸类和可溶性糖组分6大类滋味成分,以感官分析,多元统计法,结合滋味贡献度(dove-over-threshold,Dot)分析川中小叶种茶树的滋味特征。结果表明:所有茶样分为4类,Ⅰ类茶样中包含C4,C5,C6,C8以及2个对比样,各滋味成分含量相对较为均衡;Ⅱ类茶样具备类似的林下栽培环境,黄酮苷类物质含量低,没食子酸、咖啡碱(caffeine,CAFF)、儿茶素类以及氨基酸类物质含量高,表现出醇厚、鲜爽度高的滋味特征;Ⅲ类茶样为叶形较大的种质,黄酮苷类、琥珀酸为其特征滋味成分,儿茶素类、茶氨酸和CAFF含量低,涩味和鲜爽度较高;Ⅳ类茶样儿茶素类、CAFF含量较高,Thea含量低,滋味浓强,但欠鲜爽。该研究对万州区不同区域川中小叶种制备红茶的品质特性进行了梳理,为该类种质资源进一步改进工艺和开发利用提供理论依据。

The Tea Tree Genome Provides Insights into Tea Flavor and Independent Evolution of Caffeine Biosynthesis

Tea is the world＇s oldest and most popular caffeine-containing beverage with immense economic, medicinal, and cultural importance. Here, we present the first high-quality nucleotide sequence of the repeat-rich （80.9%）, 3.02-Gb genome of the cultivated tea tree Camellia sinensis. We show that an extraordinarily large genome size of tea tree is resulted from the slow, steady, and long-term amplification of a few LTR retrotransposon families. In addition to a recent whole-genome duplication event, lineage-specific expansions of genes associated with flavonoid metabolic biosynthesis were discovered, which enhance catechin production, terpene enzyme activation, and stress tolerance, important features for tea flavor and adaptation. We demonstrate an independent and rapid evolution of the tea caffeine synthesis pathway relative to cacao and coffee. A comparative study among 25 Camellia species revealed that higher expression levels of most flavonoid- and caffeinebut not theanine-related genes contribute to the increased production of catechins and caffeine and thus enhance tea-processing suitability and tea quality. These novel findings pave the way for further metabolomic and functional genomic refinement of characteristic biosynthesis pathways and will help develop a more diversified set of tea flavors that would eventually satisfy and attract more tea drinkers worldwide.

茶树油对脂多糖诱导奶牛小肠上皮细胞损伤的影响

本试验旨在研究茶树油(TTO)对脂多糖(LPS)诱导奶牛小肠上皮细胞(BIECs)损伤的影响。试验选取3头健康新生中国荷斯坦犊牛的空肠组织,分离并获得BIECs进行培养。通过使用不同浓度的LPS(0、1、2和4μg/mL)和不同浓度的TTO(0、0.00625%、0.01250%、0.02500%、0.05000%和0.10000%),建立细胞炎症模型。然后,对照组以不含TTO和LPS的培养基处理细胞;LPS组以1μg/mL的LPS处理细胞12 h;LPS+TTO组采用0.01250%和0.02500%的TTO预处理细胞12 h,经水洗后,再暴露于LPS处理12 h。结果表明:1)与对照组(未添加TTO)相比,添加0.00625%、0.01250%、0.02500%和0.05000%TTO对BIECs细胞活力无显著影响(P>0.05),添加TTO显著提高跨膜电阻(TEER)(P<0.05)。与对照组相比,添加0.01250%TTO显著提高BIECs中闭锁小带蛋白-1(ZO-1)、封闭蛋白(occludin)、闭合蛋白-1(claudin-1)和闭合蛋白-4(claudin-4)的mRNA相对表达量(P<0.05),显著降低肿瘤坏死因子-α(TNF-α)的mRNA相对表达量(P<0.05)。2)与LPS组相比,LPS+TTO组TEER以及ZO-1、occludin和claudin-1的mRNA相对表达量显著提高(P<0.05),TNF-α的mRNA相对表达量显著降低(P<0.05);LPS+TTO组ZO-1蛋白相对表达量显著提高(P<0.05)。综上可知,TTO可以通过上调紧密连接蛋白表达和下调炎性因子表达,缓解LPS诱导的犊牛小肠上皮细胞屏障功能障碍和炎症损伤。

茶树精油微胶囊制备及其结构表征

为解决茶树精油稳定性差,对光、热、氧和湿度高度敏感,挥发性较强等问题,利用复合凝聚法制备微胶囊提高茶树精油稳定性。研究优化复合凝聚法制备茶树精油微胶囊的工艺,探究不同均质时间、均质速度、pH及凝聚时间对茶树精油乳液乳化效果和微胶囊的微观形态影响,并阐明茶树微胶囊包埋特性和缓释性能。研究结果表明,以明胶-六偏磷酸钠复合凝聚层为壁材,茶树精油乳液制备条件均质转速为20000 r/min,均质时间为15 min;茶树精油微胶囊复合凝聚pH为4.37,得到形态较均匀、微粒直径为0.80~1.32μm,包埋率为(44.75±0.09)%,缓释性能为65.13%。该研究为茶树精油微胶囊的制备提供必要的数据支撑。