Journal of Tea Science

Research Advances on the Drought-Resistance Mechanism and Strategy of Tea Plant

LIU Shengchuan, CHEN Liang

Journal of Tea Science. 2014, 34(2): 111-121. doi:10.13305/j.cnki.jts.2014.02.001

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Drought is one of the pivotal meteorological disasters affecting tea production. Responses of tea plant to drought and rehydration are very complex, so it is vital to study drought-resistance mechanism for exploring relative genetic resources, improving water use efficiency and drought resistance, developing water-saving and drought-resistance cultivation and subsequent reducing losses. Here, effects of drought stress on growth, development, production and quality of tea were presented. We try to provide a brief insight into how tea plant responds to drought and rehydration. Some recent studies on strategies for water-saving and drought-resistance cultivation and subsequent recovery had been reviewed, and major research trends were discussed.

The Effect of Mixed Planting on Tea Production, Main Predators and Insect Pests in Tea Plantations

SU Hongfei, YANG Liuxia, ZHEN Wenzhong, WANG Xinghua, YAO Xuekun, CUI Tinghong

Journal of Tea Science. 2014, 34(2): 122-128. doi:10.13305/j.cnki.jts.2014.02.003

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The tea productions, main predators and insects pests of five mixed planting tea plantations were analyzed in Puer city. The effects of mixed planting on productions of fresh tea leaves, predators and insect pests in tea plantation were investigated. The results showed that predatory spiders and Empoasca vitis were most important predator and pest respectively in five tea plantations. Numbers of Empoasca vitis in mono-tea plantation were significantly higher than mixed planting plantation. The orders of spider individual numbers in five tea plantations were as follows: mixed planting of Dimocarpus longan and tea>mixed planting of Castanea mollissina and tea>mixed planting of Alstonia scholaris and tea> mixed planting of Cunninghamia lanceolata and tea> mono-tea plantation. The pests in mixed planting of Dimocarpus longan and tea plantation were significantly less than those in other tea plantations. There were no significant differences among mono-tea plantation, mixed planting of Castanea mollissina and tea plantation, mixed planting of Alstonia scholaris and tea plantation, and mixed planting of Cunninghamia lanceolata and tea plantation, mixed planting of Dimocarpus longan and tea plantation were the best mixed types because of moderate light, highest production, most predators and less pests. There were significant negative correlations between amounts of predatory spider and Empoasca vitis as well as other insect pests in different season which can produce a good representation by used of Logarithm equation.

Composition and Stability of Soil Aggregates among Different Soil Types of Tea Garden

WANG Feng, CHEN Yuzhen, YOU Zhiming, WU Zhidan, JIANG Fuying, CHEN Zhizhi, WENG Boqi

Journal of Tea Science. 2014, 34(2): 129-136. doi:10.13305/j.cnki.jts.2014.02.004

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Based on the field survey and laboratory analysis, the soil aggregate composition and stability characteristics in five soil types (yellow soil, red earth, moisture sandy soil, alpine meadow soil and purple soil) of tea garden in Wuyishan city were investigated. The results showed that the contents of soil macro-aggregates decreased with soil depth, but micro-aggregates increased correspondingly in the five soil types of tea garden. The contents of soil macro-aggregates (>5 mm) were highest, accounting for 18.07%-89.23% of the total in yellow soil, alpine meadow soil and purple soil, the soil micro-aggregates (<0.25 mm) were the highest, accounting for 37.16%-63.91% of the total in red soil and moisture sandy soil. The proportion of R0.25 (water-stable aggregate >0.25 mm ) was between 36.08% and 97.23% (sample means 73.60%). The mean weight diameter (MWD) and geometric mean diameter (GMD) of soil aggregates in alpine meadow soil of tea garden were significantly higher than those in other soil types, followed by those in yellow soil and purple soil, the lowest was in moisture sandy soil. The fractal dimension (D) changes in the opposite trend simultaneously. The soil aggregate composition and stability were significantly correlated with soil organic carbon, total N, total P, soil bulk density, soil porosity, sand and silt, but not significant correlated with C/N, total K, pH value and clay.

Evaluation of Methods for Extraction of Quality-related Constituents in Fresh Tea Leaves

YANG Yiyang, ZHANG Jia, WANG Chuanpi, LI Xinghui, RUAN Jianyun

Journal of Tea Science. 2014, 34(2): 137-144. doi:10.13305/j.cnki.jts.2014.02.005

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Metabolomic and systems biology require the acquisition of reproducible, reliable and homogeneous biological data set, the efficient extraction method is the key step in getting the satisfactory results of investigation. The influence on the extracting effect of quality-related constituents in tea leaves was investigated on various extracting solvents(methanol, ethanol, HClO₄) and various grind methods (ball grinder and mortar). Results showed that 70% ethanol and 70% methanol proved to be more suitable solvent for extracting the amino acids in fresh tea leaves compared with 100% methanol and 100% ethanol. For tea polyphenols, 100% ethanol and 70% ethanol showed the highest extraction efficiency. 3 mol·L^-1HClO₄ showed obvious advantage over all other solvents for amino acids extraction in fresh tea leaves, however, it showed the lowest extraction efficiency in tea polyphenols extraction. The use of 70% ethanol and grinded by ball grinder for 5 min has been proved to be the most suitable method. In addition, the amino acids and tea polyphenols concentration in freeze-dried leaves and oven-dried leaves were compared, the results showed that freeze-dry process could retain more quality-related constituents in tea leaves.

Preliminary Reserch of Tea Polyphenols on Insulin Resistance

XIA Jiajie, XUE Meng, JIN Meng, WU Jueli, LIU Handeng

Journal of Tea Science. 2014, 34(2): 144-148. doi:10.13305/j.cnki.jts.2014.02.006

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To study the effects of tea polyphenols (TP) on insulin resistance in the model rats of T2DM with NAFLD and its mechanism. Adult healthy SD rats were randomly divided into six equal groups: normal control (NC) group, model group, TP group, metformin group, fat liver (FL) group and FL+TP group. In the end of two weeks treatment, the levels of fasting serum glucose, insulin, TG and the TG of liver were measured. Two weeks after treatment, the fasting blood glucose and insulin and insulin sensitivity index of the TP group and metformin group were both significantly lower than those of the model group (both P<0.05). The TP group and FL+TP group compared with model group and FL group, the TP significantly reduce the levels of TG of the liver (both P<0.05). TP can efficiently ameliorated lipid metabolism in the liver tissue of rats and improve insulin sensitivity in the rat model of type 2 diabetes mellitus with fatty liver.

The Antimicrobial Mechanism of Tea Polyphenols Against Shewanella putrefaciens

WANG Huimin, ZHU Junli, LU Haixia, YI Shumin

Journal of Tea Science. 2014, 34(2): 149-155. doi:10.13305/j.cnki.jts.2014.02.008

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Antimicrobial effect and mechanism of TP against Shewanella putrefaciens in vitro was investigated. The results showed that the minimum inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of TP against the S. putrefaciens were 1 mg·mL^-1 and 16 mg·mL^-1, respectively. Growth curves of S. putrefaciens were significantly inhibited in the addition of 2 mg·mL^-1 TP. The population of S. putrefaciens decreased about 10⁵ CFU·mL^-1 and 10⁶ CFU·mL^-1 with the 2 mg·mL^-1 TP addition after 96 h at 25℃ and 192 h at 4℃, respectively. The bactericidal activity of TP exhibited the time depending. Among the four monomer of TP, the order of bactericidal activity were EGCG>ECG>EGC>EC. The enzyme activity of Na⁺/K⁺-ATP and AKP in the culture suspension of S. putrefaciens gradually increased with the time extension in the TP treated samples. TEM images revealed that treatment of TP might lead to damage and deformation of cell. Thus, TP exhibited the good antimicrobial and bactericidal activity against S. putrefaciens by disrupting cell walls and out membrane, which cause the permeability of intracellular components and the change of cell morphology.

Study on the Inhibition Effects of EGCG on the Expression of RCCs-RAGE Axis in Diabetic Mouse Induced by Alloxan Monohydrate

ZHONG Yuan, ZHANG Jing, LIU Zhonghua, CAI Shuxian, LUO Guoan, HUANG Jian′an, WU Xianglan, QU Shaoming, JIN Lisha

Journal of Tea Science. 2014, 34(2): 156-164. doi:10.13305/j.cnki.jts.2014.02.009

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The RCCs-RAGE signal axis induced by RCCs, including AGEs and 4-HNE, plays a key role in degenerative diseases, such as diabetes and cancer. To study on inhibiting activity effects of EGCG on RCCs-RAGE, diabetic mouse induced by intraperitoneal injection Alloxan monohydrate were divided into model group, EGCG-L (10 mg·kg^-1·d^-1) group, EGCG-M (20 mg·kg^-1·d^-1) group, and EGCG-H (30 mg·kg^-1·d^-1) group according to body weight and blood glucose. After 12 days’ administration respectively to three groups of diabetic mouse by gavage, blood glucose value, insulin, sRAGE concentration, carbonyl content and fluorescence value of AGEs in serum were determinde, while the expressions of RAGE gene in kidney were detected by QPCR, RAGE protein and 4-HNE experiment by western bolt. Result showed, compared to model group, EGCG could improve diabetes symptom of mouse by significantly decreasing the formation of RCCs, including carbonyl, 4-HNE and AGEs, increasing the concentration of sRAGE in serum, inhibiting the inflammation reaction induced by RCCs-RAGE, and alleviating the oxidation stress effectively. The study revealed that EGCG inhibited the RCCs-RAGE signal axis may be one of the potential mechanisms in the treatment of degenerative diseases.

Molecular Binding Characterization with Tea Plant Volatiles of a General Odorant-binding Protein EoblGOBP2 in the Tea Geometrid, Ectropis oblique Prout (Lepidoptera: Geometridae)

ZHAO Lei, CUI Hongchun, ZHANG Linya, CHEN Ling, YU Jizhong, LI Hongliang

Journal of Tea Science. 2014, 34(2): 165-171. doi:10.13305/j.cnki.jts.2014.02.010

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As one main Lepidopteran pest in tea garden, the larvae of tea geometrid damages mainly the tea leaves, which severely reduce the yield of tea products. The olfactory system of tea geometrid plays a very important role in terms of seeking host and food. Therefore, a general odorant binding protein—EoblGOBP2, existing in the olfactory system of tea geometrid, the function binding with tea plant volatiles was studied in vitro here. Based on the cloned EoblGOBP2 gene by RT-PCR, the recombinant protein EoblGOBP2 was successfully expressed in an optimized prokaryotic prokaryotic expression system, then the binding characteristics of the purified EoblGOBP2 with 1-NPN was measured by fluorescence spectrometry. Scatchard plot analysis indicated that the dissociation constant between EoblGOBP2 and 1-NPN was 2.310 μmol·L^-1. In the competitive binding assays, to the seven candidate ligands, all of the relative fluorescence intensity of 1-NPN decreased more than 50%, which suggested that all of the tested ligands have strong binding capability with EoblGOBP2. The compound of dibutyl phthalate showed the maximum binding capability with EoblGOBP2, and their dissociation constant was 4.353 μmol·L^-1. As all of the candidate chemical ligands belong to the tea volatiles secondary metabolites, indicating that EoblGOBP2 plays an important role in the odor recognition process of olfactory identification in the olfactory system of tea geometrid.

Comparison of Growth Period and Biochemical Composition on Fujian Tea Cultivars and Genetic Diversity Analysis

CHEN Changsong, CHEN Zhihui, ZHONG Qiusheng, LIN Zhenghe, YOU Xiaomei, WANG Xiuping, YU Wenquan

Journal of Tea Science. 2014, 34(2): 172-179. doi:10.13305/j.cnki.jts.2014.02.012

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The spring phenophase, biochemical composition and genetic diversity analysis of the 40 tea cultivars with same age, which passed the national and provincial examination or approval from Fujian, were surveyed and identified under the same ecological and cultural condition for two years. The results showed that among the 26 tested oolong tea cultivars, 31% of those were early sprouting cultivars, 23% were late sprouting cultivars and 46% were medium or medium-late sprouting cultivars, among the tested 14 green tea cultivars, 29% of those were extra-early sprouting cultivars, 7% were late sprouting cultivars, and the rest 64% were early or medium sprouting cultivars. Significant differences on bud stage among tea cultivars provided a strong basis for reasonable collocation on bud stage in production. In the tested cultivars, water extracts was from 40% to 52%, tea polyphenols from 9.8% to 20.8%, free amino acids from 3% to 6.8%, and caffeine from 2.9% to 5.0%. The results showed that the contents of each biochemical component among different cultivars had a relatively significant difference. The genetic diversity of tested cultivars was relatively higher, genetic distances of Zaochunhao, Zhenghe-dabaicha, Xiapu-yuanxiaocha, and Yuemingxiang were relatively farther away, and the rest 36 cultivars could be divided into 4 main groups: Tieguanyin type, Minnan type, Fuding-dabaicha type, and Huangdan type. This tudy provided important data reference and theoretical guidance for diversity of choice in production cultivars.

Study of SNaPshot Detect SNP Markers in Tea Plant

ZHANG Chengcai, TAN Liqiang, WANG Liyuan, WEI Kang, CHENG Hao

Journal of Tea Science. 2014, 34(2): 180-187. doi:10.13305/j.cnki.jts.2014.02.014

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In order to increase the genotyping efficiency of tea SNPs and promote the application of SNPs in tea genetic breeding investigation, the feasibility of SNaPshot in SNPs genotyping of tea plant was investigated. Ten SNPs were selected from previous experiment as target SNPs. Then, these SNPs were detected in different tea cultivars by SNaPshot technology. Six among 10 SNPs were successful detected, with an accuracy of 60%. The polymorphism diversity of these SNPs was also analyzed. The value of N_A was 2, H_e ranged from 0.37 to 0.52, H_o ranged from 0.32 to 0.74, PIC ranged from 0.36 to 0.50. All these markers were shown coincide between `Longjing43` and its repeated test. The markers reported here will be useful for tea genetic linkage map construction and genetic diversity study. The SNaPshot technology will promote the genetic study and accelerate the breeding process in tea plant.

Cloning and Sequence Analysis of the N-methyltransferase Gene Family Involving in Caffeine Biosynthesis of Tea Plant

JIN Jiqiang, YAO Mingzhe, MA Chunlei, MA Jiangqiang, CHEN Liang

Journal of Tea Science. 2014, 34(2): 188-194. doi:10.13305/j.cnki.jts.2014.02.015

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Caffeine is an important bioactive compound in tea. It is synthesized from xanthosine catalysed by N-methyltransferase (NMT) that using S-adenosylmethionine (SAM) as the methyl donor. Based on the characterization of high similarity of CS (Caffeine Synthase) gene sequences, 6 NMTs were cloned from Baiye 1 by using the technique of long PCR and cloning flanking DNA sequence. Two of them are TCS1 and TCS2, one is a pseudogene, and the other three are named TCS3, TCS4 and TCS5, respectively. It was found that these genes were all composed by four exons and three introns through analysis of gene structure. The NMTs of Camellia plants were clustered into 5 groups, the similarity of TCS4 and TCS5 between with other genes is lower. These results provided useful references to dissect the genetic mechanism of caffeine from genome level in tea plant for the future.

Analysis of Differential Gene Expression on Flavonoid Biosynthesis and Its Regulation in Tea Plant (Camellia sinensis L.)

WEI Kang, WANG Liyuan, CHENG Hao, ZHANG Liqun, FENG Suhua

Journal of Tea Science. 2014, 34(2): 195-203. doi:10.13305/j.cnki.jts.2014.02.016

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The expression of genes associated with flavonoid biosynthesis and its regulation in petal, pollen, dormant and sprouting buds of tea plants were analyzed by differential gene expression. The results showed that flavonoid biosynthetic genes were highly expressed in petal, dormant and sprouting buds, but rarely expressed in pollen, which is consistent with the distribution of flavonoids in tea plant. Furthermore, twelve genes in MYB, bHLH, MADS, GST, WD40 and Homeodomain gene families were identified to be correlated with flavonoid biosynthesis in petal. Nine genes were found to be involved in the regulation of flavonoid biosynthesis in tea buds. These results will be a basis for further understanding the mechanism of flavonoid biosynthetsis and regulation in Camellia sinensis.

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