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.

Comparative Effect of Ca, K, Mn and B on Post-Drought Stress Recovery in Tea [<i>Camellia sinensis</i>(L.) O <i>Kuntze</i>]

Tea (Camellia sinensis) is one of the most important economic crops. Being perennial in nature, tea plant often experiences natural drought, which affects its growth and productivity. The present investigation was undertaken to understand the mechanism of post-drought stress recovery on rehydration and the effect of nutrients in the recovery process of the selected clones of Camellia sinensis L. (TV-1, TV-20, TV-29 & TV-30). The results demonstrated that decrease in relative water content (RWC), dry mass of leaf and antioxidants like-ascorbate and glutathione in all the tested clones, as a result of imposed water stress, which caused damage was not permanent. Increase in phenolic content with decrease in O2-, H2O2 and lipid peroxidation was indication of the recovery of stress induced oxidative damage following the post stress rehydration. Further, the post drought recovery was enhanced by foliar spray of K, Ca, Mn & B. Differential activities of enzymes like SOD, CAT, POX, GR and PPO in response to foliar spray of nutrients in rehydrated plant improved the recovery process. The present study reveals that the tested nutrients (K, Ca, Mn & B) showed some positive response in influencing growth and antioxidative responses during post drought recovery process, where K and Ca showed comparatively better effect in improving post drought recovery potential in tea plant.

Quantitative Analysis of ATP Sulfurylase and Selenocysteine Methyltransferase Gene Expression in Different Organs of Tea Plant (<i>Camellia sinensis</i>)

Tea plant (Camellia sinensis) has unique biological features for the study of cellular and molecular mechanisms, an evergreen broad-leaved woody plant which can accumulate selenium in soil abundant of Selenium. Expression of the genes related to Selenium (Se) metabolism is an adaptation to the soil environment for a long period. The purpose of the present study was to explore if there exist differences of expression about these genes in tea plant between growing in Selenium-abundant and normal soil. A quantitative real-time reverse transcription polymerase chain reaction (Q-RT-PCR) assay was done for quantification of ATP sulfurylase (APS) and selenocysteine methyltransferase (SMT) mRNA normalized to Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene in tea plant. Young leaves, mature leaves and tender roots from tea plants growing in soil abundant of Selenium were respectively obtained from Shitai County, Anhui Province, and also the relevant materials of the selenium un-enriched tea plant planted at agricultural garden of Ahui Agriculture University were taken as control for real-time PCR analysis. The results showed that APS1, APS2 and SMT expression levels for either young or mature leaves in selenium-enriched tea plant were lower than that in ordinary (selenium un-enriched) tea plant. In contrast, the APS1, APS2 and SMT expression level of roots in selenium-enriched tea plant were all higher than that in ordinary tea plant. APS1 gene expression level of roots in selenium-enriched tea plant was about 1.6 times higher than that in the ordinary tea plant, APS2 gene expression level was about 4.8-fold higher than that in the ordinary tea plant, SMT gene expression level was about 3.3 times higher than that in the ordinary tea plant. Among various tissues of selenium-enriched tea plant, APS1 gene relative expression level of young leaves was similar to or slightly higher than mature leaves, and the one of roots was the lowest among them;APS2 gene relative expression level of young leaves was similar to or slightly higher than the roots, and the one of mature leaves was the lowest among them;SMT gene relative expression level of young leaves was similar to or slightly higher than mature leaves, and the one of roots was the highest among them. Our results suggest that there existed correlation between selenium and expression levels of these genes.

Identification of miRNAs and target genes regulating catechin biosynthesis in tea (Camellia sinensis)

Micro RNAs（miRNAs） are endogenous non-protein-coding small RNAs that play crucial and versatile regulatory roles in plants. Using a computational identification method, we identified 55 conserved miRNAs in tea（Camellia sinensis） by aligning miRNA sequences of different plant species with the transcriptome library of tea strain 1005. We then used quantitative real-time PCR（qRT-PCR） to analyze the expression of 31 identified miRNAs in tea leaves of different ages, thereby verifying the existence of these miRNAs and confirming the reliability of the computational identification method. We predicted which miRNAs were involved in catechin synthesis using ps RNAtarget Software based on conserved miRNAs and catechin synthesis pathway-related genes. Then, we used RNA ligase-mediated rapid amplification of c DNA ends（RLM-RACE） to obtain seven miRNAs cleaving eight catechin synthesis pathway-related genes including chalcone synthase（CHS）, chalcone isomerase（CHI）, dihydroflavonol 4-reductase（DFR）, anthocyanidin reductase（ANR）, leucoanthocyanidin reductase（LAR）, and flavanone 3-hydroxylase（F3H）. An expression analysis of miRNAs and target genes revealed that miR529d and miR156 g-3 p were negatively correlated with their targets CHI and F3H, respectively. The expression of other miRNAs was not significantly related to their target genes in the catechin synthesis pathway. The RLM-RACE results suggest that catechin synthesis is regulated by miRNAs that can cleave genes involved in catechin synthesis.

Induced Polyploidy as a Tool for Increasing Tea(Camellia sinensis L.) Production

Tea （Camellia sinensis L.） represents different ploidy levels. In the present paper, we reviewed the recent data on the diploid, aneuploid and polyploid formed their origin and chemically induced polyploidy and predicted it role in teagenetic improvement for better yield. Different polidy levels had different effects on tea physiology. Tetraploid and triploid had more vigour and hardness due to increased size of cells, while triploid could have even more vigours due to increased size of cells and sterilities. Chemically induced polyploidy had been found an important tool for improving plant physiology and production, therefore, induced polyploids should be produced to overcome the problem of low yield and limited rainfall in tea growing areas.

Peroxidase Inhibition and Antioxidant Activity of Bulk-Marketed Black Tea (<i>Camellia sinensis</i>L.) from the Democratic Republic of the Congo

The widely known leaves of Camellia sinensis (L.) derive the most consumed beverage globally. All over the world, like in the Democratic Republic of the Congo (DR Congo), they are marketed in the form of packaged and in bulk black tea. The aim of this study was to report the microscopic botanical features, the chromatographic fingerprints, the antioxidant activities, and the peroxidase inhibition of bulk-marketed black teas consumed in DR Congo. The microscopic analysis of powders from unpackaged black tea marketed in DR Congo allowed the identification of specific microscopical features, which are characteristic to C. sinensis (L.) such as numerous asterosclerites, numerous fragments of unicellular trichomes;fragments of abaxial epidermis consisting of cells with slightly wavy walls, numerous anomocytic stomata, and the detection of biological contaminants, e.g. the dust mites. TLC and HPLC analysis indicated that flavonoids (hyperoside, rutin) and phenolic acids (chlorogenic acid, gallic acid) were major phenolic compounds, along with other secondary metabolites. Aqueous extracts from all bulk-marketed black teas exhibited good antioxidant activities at the concentrations range of 1 - 20 μg·mL-1 using in vitro ABTS and DPPH assays. These extracts showed equally inhibitory effects on myeloperoxidase (MPO) and horseradish peroxidase (HRP) activities at the concentration ranges of 1 - 20 μg·mL-1 and 20 - 100 μg·mL-1 respectively. Bulk-marketed black teas consumed in DR Congo possess antioxidant and anti-inflammatory properties comparable to packaged Ceylon tea largely marketed. The observed bioactivities support the health benefits of the consumed black tea by Congolese people, despite the storage conditions to be improved in order to preserve its nutritional and therapeutic assets.

In vitro anti-hyaluronidase activity of Sri Lankan low grown orthodox orange pekoe grade black tea(Camellia sinensis L.)

Objective:To access the anti-hyaluronidase activity of Sri Lankan low grown orthodox orange pekoe(OP) grade black tea with a view to develop an anti-aging skin formulation.Methods:Five concentrations(0.125,0.250,0.500,1.000 and 2.000 mg/mL) of black tea brew(BTB) were made using a freeze dried sample of Sri Lankan low grown orthodox OP grade black tea which was prepared according to international organization for standardization specification.Epigallocatechin gallate(EGCG) was used as the reference agent(concentrations tested:0.012,0.025,0.050,0.100 and 0.200 mg/mL).Anti-hyaluronidase activity of BTB and EGCG in vitro were ascertained spectrometrically using hyaluronic acid(from rooster comb)and bovine testicular hyaluronidase.Results:The results revealed that BTB had moderate[IC_(50)=(1.09±0.12) mg/mL]and dose dependent(r^2=0.94) anti-hyaluronidase activity.EGCG also exhibited dose dependent(r^2=0.93,P<0.05) anti-hyaluronidase activity which was superior[IC_(50)=(0.09±0.00) mg/mL]to BTB.Conclusions:Sri Lankan low grown orthodox OP gradc black tca has promising antihyaluronidase activity in vitro and has the potential to be used as an anti-aging cosmaceutical.In addition,it may prove useful as a beverage in the management of allergy,some joint diseases and cnvenomation.

Screening of Adoptive Elite Tea (Camellia sinensis) Clones

The research screening of adoptive elite tea clones was conducted at NTRI, Mansehra during 2011-2012. Nine clones 101Aa, 105aa, 108aa, 561aa, ll7aa, 219ab, 470bb and 180bd were evaluated for seedling performance. Randomized complete block design was used with three replications. Data was recorded on various morphological characters after 8 months. The results showed that high survival percentage, shoot length, number of roots plant-1, number of leaves plant1 and root length were observed in clone 105aa. While the highest fresh weight and dry weight of leaves were observed in clones 117aa and 105aa. The clone 105aa was drought resistant, high survival percentage and root growth. On the basis of the results, clone 105aa was recommended for cultivation through cuttings in the hilly areas of Pakistan where unequal rainfall distribution was a major hitch.

Genome-Wide Characterization of the Cellulose Synthase Gene Superfamily in Tea Plants(Camellia sinensis)

The cellulose synthase gene superfamily,including Cellulose synthase A(CesA)and cellulose synthase-like(Csl)gene families,is responsible for the synthesis of cellulose and hemicellulose,respectively.The CesA/Csl genes are vital for abiotic stress resistance and shoot tenderness regulation of tea plants(Camellia sinensis).However,the CesA/Csl gene family has not been extensively studied in tea plants.Here,we identified 53 CsCesA/Csl genes in tea plants.These genes were grouped into five subfamilies(CsCesA,CsCslB,CsCslD,CsCslE,CsCslG)based on the phylogenetic relationships with Arabidopsis and rice.The analysis of chromosome distribution,gene structure,protein domain and motif revealed that most genes in CsCesA,CsCslD and CsCslE subfamilies were conserved,whereas CsCslB and CsCslG subfamily members are highly diverged.The transcriptome analysis showed that most CsCesA/Csl genes displayed tissue-specific expression pattern.In addition,members of CsCslB4,CsCesA1/3/6,CsCslB3/4,CsCslD3,CsCslE1 and CsCslG2/3 subfamilies were up-regulated under drought and cold stresses,indicating their potential roles in regulating stress tolerance in tea plants.Furthermore,the expression levels of CsCslG2_6 and CsCslD3_5 in different tissues and cultivars,respectively,were positively correlated with the cellulose content that is negatively related with shoot tenderness.Thus,these two genes were speculated to be involved in the regulation of shoot tenderness in tea plants.Our findings may help elucidate the evolutionary relationships and expression patterns of the CsCesA/Csl genes in tea plants,and provide more candidate genes responsible for stress tolerance and tenderness regulation in tea plants for future functional research.

Functional Components of Tea(Camellia Sinensis)Germplasms Collected in Korea

This study was carried out to obtain useful information and to search quality related components for breeding of tea by analysis of Korean local tea leaves at Sulloc-Cha R&D center in Jeju Island,Korea.It was also conducted to measure the major agronomic characteristics of 300 local germplasms.These results allowed us to select some superior lines as genetic resources for green-tea breeding and research genotypes.

茶(Camellia sinensis L.)对铜的吸收与累积

对广东省5个大型茶场中不同土壤背景值的8个茶园进行了土壤及茶(CamelliasinensisL.)树各部位的铜含量分析调查。地处我国南方的广东省茶园,土壤中铜含量在2.39～53.05mg kg之间。有效态铜含量相对较低,仅在0.45～3.17mg kg之间。茶树中各部位铜含量大小依次为枝条>幼叶>根>成熟叶。茶树枝条和幼叶铜含量较高,平均分别达14.5和14.3mg kg,远高于成熟叶铜含量水平(9.7mg kg)。而同时期相应茶场的茶叶制成品铜含量则较幼叶(一芽两叶)铜的含量高。土壤中的总钾、有效磷和总铜等都对茶树各部位铜的累积产生较大影响,但只有土壤有效磷含量对铜在茶树体内向枝条部分的转移和累积影响达显著程度,枝条铜含量与土壤有效磷之间的相关系数r=-0.74,呈显著负相关(P<0.05)。

茶(Camellia sinensis)蜂花粉及其蜂粮的营养成分研究

测定中国茶 (Camellia sinensis)的新鲜蜂花粉、5 d、1 0 d、1 5 d茶蜂粮中水解氨基酸的含量 ,茶蜂粮中平均含量为2 3 .3 6% ,比新鲜茶蜂花粉高 0 .2 6% ;新鲜茶蜂花粉和混合 (5～ 1 0 d)茶蜂粮样品中游离氨基酸含量分别为 1 0 3 7.46mg/1 0 0 g和 80 1 .75 mg/1 0 0 g。混合 (5～ 1 0 d)茶蜂粮和新鲜茶蜂花粉样品中维生素 A、B1 、B2 、C、D、E的含量 (mg/1 0 0 g)分别是0 .1 9、0 .1 5、1 .1 5、0 .3 1、0 .0 1 5 6、1 0 .2和 0 .79、0 .0 9、2 .74、1 .2 0、0 .0 1 5 8、6.6。新鲜茶蜂花粉和混合茶蜂粮样品中软脂酸、硬脂酸、亚油酸、亚麻酸的含量分别为 1 .3 %、0 .2 %、0 .1 6%、2 .5 %和 1 .1 %、1 .0 %、0 .3 %、1 .0 %。1 d茶蜂粮中的过氧化氢酶、淀粉酶的活力比新鲜茶蜂花粉高 ,而过氧化物歧化酶的活力则比新鲜茶蜂花粉略低 ;随着酿制时间的延长 ,茶蜂粮中过氧化氢酶、过氧化物歧化酶的活力下降 。

茶(Camellia sinensis)花粉及其蜂粮中花粉粒的显微结构

采用光学显微镜和扫描电镜观察茶蜂花粉及其不同酿制时间的蜂粮中花粉粒的外部形态和萌发沟,未发现花粉壁和花粉粒结构被破坏的迹象.通过体外模拟消化试验,采用光学显微镜和透射电镜观察,发现经模拟消化的蜂粮中花粉粒内含物外吐的数量比蜂花粉多,而且随着酿制时间的延长,其内含物外吐的花粉粒所占的比例增加.结果提示,同种粉源的蜂粮比蜂花粉更易于被消化.

外源24-表油菜素内酯诱导茶树(Camellia sinensisL.)耐热性的生理机制

以茶树品种龙井43为试验材料,研究外源24-表油菜素内酯(EBR)处理对高温胁迫下茶树叶片光合特性、叶绿素荧光参数、丙二醛(MDA)含量和抗氧化酶活性的影响。研究发现:叶面喷施EBR能显著提高高温胁迫下茶树叶片的净光合速率(Pn),气孔导度(Gs),Rubisco最大羧化速率(V_(c,max))、Rubisco的最大再生速率(J_(max))和PSⅡ最大光化学效率(F_v/F_m);同时外源EBR处理后,高温胁迫下茶树叶片中的抗坏血酸过氧化物酶(APX)、过氧化氢酶(CAT)、超氧化物歧化酶(SOD)和过氧化物酶(POD)活性显著提高,而MDA含量则显著下降。外源EBR主要通过解除非气孔因素的限制、促进光合碳反应的进行来缓解高温胁迫对茶树叶片光合作用的抑制;同时EBR能够有效利用多种抗氧化途径以清除其细胞内的自由基和活性氧,最终缓解高温胁迫对茶树的伤害。

Promoter and coding sequence diversity of CsCCD1 may contribute to the differential accumulation of floralβ-ionone in fresh tea leaves

The carotenoid-derived volatileβ-ionone makes an important contribution to tea fragrance.Here,we qualitatively and quantitatively analysed 15 carotenoids in tea leaves of 13 cultivars by UHPLC-APCI-MS/MS.The 13 cultivars were divided into two groups by PCA(Principal Component Analysis)clustering analysis of their carotenoid content,and OPLS-DA(Orthogonal projections to latent structures)indicated that the levels ofβ-carotene(VIP=2.89)and lutein(VIP=2.30)were responsible for much of the variation between the two groups.Interestingly,theβ-carotene toβ-ionone conversion rates in Group 1 were higher than in Group 2,while theβ-carotene content was significantly lower in Group 1 than in Group 2.Theβ-ionone content was significantly higher in Group 1.Pearson Correlation Coefficient calculation between the transcription level of candidate genes(CsCCD1 and CsCCD4)and the accumulation ofβ-ionone indicated that CsCCD1 may involve in the formation ofβ-ionone in 13 cultivars.Prokaryotic expression and in vitro enzyme activity assays showed that‘Chuanhuang 1’had an amino acid mutation in carotenoid cleavage dioxygenases 1(CsCCD1)compared with‘Shuchazao’,resulting in a significantly higherβ-ionone content in‘Chuanhuang 1’.Sequence analysis showed that‘Chuanhuang 1’and‘Huangdan’had different CsCCD1 promoter sequences,leading to significantly higher CsCCD1 expression andβ-ionone accumulation in‘Chuanhuang 1’.These results indicated that the promoter and coding sequence diversity of CsCCD1 might contribute to the differential accumulation ofβ-ionone in different tea cultivars.

大叶茶(Camellia sinensis var. assamica)的命名、模式及自然分布

大叶茶(Camellia sinensis var. assamica)在全球广泛种植,是商品茶叶的重要原料来源。本文系统地回顾了大叶茶学名的合格发表及其命名模式的研究进展。依据《深圳法规》条款41.4,Griffith在1854年将茶组[Camellia sect. Thea (L.) Griff.]作为新等级名称、而非新类群名称合格发表。老挝茶因其叶背具腺点、花柱离生而被移出茶组。Masters在1844年并未合格发表Thea assamica,此名称最早由Hooker于1847年合格发表,Steenis于1949年提出的新组合Camellia sinensis (L.) Kuntze var. assamica (Hook.) Steenis应为大叶茶的完整学名。英国邱园馆藏标本W. Griffith s.n.(K000939670)在2021年被指定为大叶茶的新模式。本文同时总结了迄今归并的7个大叶茶的异模式异名的原白和模式。尽管Darlington和Ammal在1945年将大叶茶作为物种处理并提出新组合名称C.assamica,多学科证据却支持大叶茶是茶的变种。假如大叶茶的物种地位被接受,那么1838年发表的大叶茶的异模式异名C. theifera将取代C. assamica而获得物种阶元上的优先权,届时,广泛使用的种加词“assamica”或可依据《深圳法规》条款14提议作为保留名称。本文同时总结了大叶茶在中国、印度、老挝、缅甸、泰国和越南的自然分布点,并讨论了它在不同原产国的自然资源的保护和利用现状。

Physiological Foundations of Sustainability <i>Camellia sinensis</i>(L.) O. Kuntze and <i>Corylus pontica</i>C. Koch. in the Conditions of Humid Subtropics of Russia

The study of the water status and activity of catalase in Chinese tea plants (Camellia sinensis (L.) O. Kuntze) and hazelnut (Corylus pontica C. Koch.). The indicators, which not only describe the physiological state of plants in the stressful period, but also allow it to identify more adaptive varieties within a given area was determined. Analysis of catalase activity data in Chinese tea and hazelnut revealed the presence of similar patterns: there is a change in enzymatic activity in response to hydrothermal factors. It is established that the stable varieties Camellia sinensis (L.) O. Kuntze and Corylus pontica C. Koch. characterized by a more active catalase, which is of particular importance during droughts. The water regime parameters variance scale for Camellia sinensis was developed, which allows differentiating varieties into groups of varying degrees of stability. It is shown that all methods that are used to estimate stability of plant species to extreme environmental conditions are based on several basic principles connected with peculiarities of adaptation mechanisms. The result was a selection of diagnostic indicators proposed for assessing adaptability: the water content of the leaf tissue, water-holding capacity of leaves, and concentration of cellular juice sprouts and activity of catalase.

Antimicrobial activity of green tea extract against isolates of methicillin-resistant Staphylococcus aureus and multi-drug resistant Pseudomonas aeruginosa

Objective:To evaluate antibacterial activity of the Indonesian water soluble green tea extract,Camellia sinensis,against clinical isolates of methicillin-resistantStaphylococcus aureus (S.aureus)(MRSA)and multi-drug resistant Pseudomonas aeruginosa(MDR-P.aeruginosa).Methods:Antimicrobial activity of green tea extract was determined by the disc diffusion method and the minimum inhibitory concentration(MIC)was determined by the twofold serial broth dilutions method.The tested bacteria using in this study were the standard strains and multi-drug resistant clinical isolates of S.aureus and P.aeruginosa,obtained from Laboratory of Clinical Microbiology,Faculty of Medicine,University of Indonesia.Results:The results showed that the inhibition zone diameter of green tea extracts forS.aureus ATCC 25923 and MRSA were(18.970依0.287)mm,and(19.130依0.250)mm respectively.While the inhibition zone diameter forP.aeruginosa ATCC 27853 and MDR-P.aeruginosawere(17.550依0.393)mm and(17.670依0.398)mm respectively.The MIC of green tea extracts againstS.aureus ATCC 25923 and MRSA were 400μg/mL and 400μg/mL,respectively,whereas the MIC for P.aeruginosa ATCC 27853 and MDR-P.aeruginosawere 800μg/mL,and 800μg/mL,respectively.Conclusions:Camellia sinensisleaves extract could be useful in combating emerging drug-resistance caused by MRSA andP.aeruginosa.