Journal of Tea Science

A Sequence Analysis and Structural Prediction on Polyphenol Oxidase in Tea

PAN Yu, WANG Kun-bo, XU Zhong-xi, XU Hui, HE Zhi-dan

Journal of Tea Science. 2008, 28(3): 157-165. doi:10.13305/j.cnki.jts.2008.03.003

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In the present study, Polyphenol Oxidase (PPO) in tea plant which was registered in biodatabase (DDBJ & Uni-Prot) were analyzed and predicted by method of bioinformatics, including composition of nuclei acid sequences and amino acid sequences, signal peptide, transmembrane topological structure, hydrophobicity or hydrophilicity, secondary and tertiary structure of protein. The results showed that PPO in tea was a hydrophilic and non-transmembrane protein without signal peptide, but had coils. The main motifs of its predicted secondary structure of had random coil, alpha helix and beta turn were spread to the whole secondary structure of protein. Its amino acid sequences included two functional domains.

Study on Chemical Modification and Anticoagulant Function in vitro of Tea Polysaccharides

LIANG Jin, ZHANG Jian-yun, CUI Ying-ying, HUANG Long-quan

Journal of Tea Science. 2008, 28(3): 166-171. doi:10.13305/j.cnki.jts.2008.03.010

Abstract ( 536 )

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The polysaccharides components of Olong tea were isolated and purified with DEAE-52 cellulose column chromatography. Sulfation, acetylation and carboxymethylation modification were separately selected to study on anticoagulant function in vitro of purified tea polysaccharides. The results showed that tea polysaccharides possess anticoagulant function, which could effectively prolonged APTT of human plasma, but had no obvious effect on PT and TT. Four polysaccharide components were separated by column, the component Ⅱ occupied 57.36% of total tea polysaccharides and it was relatively high anticoagulant activity. The anticoagulant function of tea polysaccharides component Ⅱ was further enhanced by sulfation,acetylation and carboxymethylation modification. The proportion between chemical modification reagents and polysaccharides, and modificating reaction time influenced on the modification degree of molecular structure of tea polysaccharides, and changed the anticoagulant activity accordingly.

Systematic Separation, Purification of Water-soluble Green Tea Polysaccharides and Their Immunological Activities

KOU Xiao-hong, JIANG He-yuan, CUI Hong-chun, ZHANG Jian-yong, GAO Qing-qing, YUAN Xin-yue, SHU Ai-min, LIU Xiao-hui, GAO Qi

Journal of Tea Science. 2008, 28(3): 172-180. doi:10.13305/j.cnki.jts.2008.03.011

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Water-soluble green tea polysaccharides were separated and purified by Ultrafiltration membranes with pore diameter of 150 kD, 20 kD, 6 kD in turn. All the three fractions were further separated and purified by DEAE-cellulose ion-exchange chromatography and Sephacryl S-300 column chromatography, and more than 20 green tea polysaccharide components totally were obtained. Their purity and molecular weight distribution were determined with high performance gel permeation chromatography (HPGPC) equipped with ELSD detector, and five components of homogeneous polysaccharide were obtained. The immunological activities of these green tea polysaccharides were determined by stimulated phagocytosis and nitric oxide production on RAW264.7 macrophage cell line of mice. Results showed that the tea Polysaccharides with 20 kD MW possessed significant immunological activities on NO production or phagocytosis of mice macrophage Raw 264.7 cells.

Study on the Change of the Main Biochemical Components, Representative Aromatic Compounds as well as Their Correlation of Green Tea Beverages during Storage

DOU Hong-liang, LI Chun-mei, HAO Ju-fang, HU Wan-feng

Journal of Tea Science. 2008, 28(3): 181-188. doi:10.13305/j.cnki.jts.2008.03.001

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The change of the main biochemical and aroma components of green tea beverage as well as their correction were studied. Results showed that the main biochemical components of green tea beverage changed significantly during storage. The relative concentration of total polyphenols decreased 26.6% after 12 months storage. After storing for 6 and 12 months, about 57.2% and 91.9% of EGCG was lost respectively. During storage, H₂O₂ was produced in the beverage, and the concentration of H₂O₂changed regularly. After storage for one month, about 68% of amino acid was lost. However, it was increased after 4th month of storage. The relative contents of the representative aromatic compounds including Linalool, Linalol oxide, Geraniol, β-Lonone, Hexanal, Benzaldehyde and Nerol in the beverage increased dramatically with the storage process. By using the sas software, the correlation analysis showed that change of the main biochemical components and the change of aroma were positively correlated. The oxidation of catechin and the change of H₂O₂ lead to the increase in concentration of Linalool, Linalol oxide, Geraniol, β-Lonone, Benzaldehyde and Nerol. The change of amino acid and H₂O₂ lead to the increase of Hexanal, besides, the oxidation of catechins was confirmed as a dominant factor which lead to the change of aroma in tea beverage during storage by multiple gradually regression analysis method.

Studies on Polyphenol-resisting Characteristics of Several Kinds of Agrobacterium and the Demestication of Polyphenol-resisting Strain

CANG Mei-qin, ZHOU Jian, CHENG Hao, LI Xing-hui, WANG Li-yuan

Journal of Tea Science. 2008, 28(3): 189-194. doi:10.13305/j.cnki.jts.2008.03.004

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In order to make clear the variation of polyphenol-resisting characteristics of different kinds of Agrobacterium as well as the potential in improving the polyphenol-resisting ability and the Agrobacterium-mediated genetic transformation rate by domestication culture, the inhibition effect of tea polyphenols on four kinds of Agrobacterium strains, the polyphenol-resisting demestication of Agrobacterium strains and the improvement of genetic transformation ability in demesticated polyphenol-resisting strain were studied. The results showed that tea polyphenols had obvious inhibition on the growth of different kinds of Agrobacterium, but it varied greatly among these studied Agrobacterium. A polyphenol-resisting strain EHA105AP was obtained by domestication culture procedure, the polyphenol-resisting ability of which enhanced 90.89% compared with that of the original. Meanwhile, the rate of transient expression of GUS gene induced by the domesticated strain increased 43.85% and 57.55% respectively in callus and leaf explants case.

Comparison on Research Methods for Fluoride Uptake Kinetics of Tea Plant

ZHANG Lei, RUAN Jian-yuan

Journal of Tea Science. 2008, 28(3): 195-200. doi:10.13305/j.cnki.jts.2008.03.005

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Tea plant (Camellia sinensis L) was employed as a model plant to study fluoride (F) uptake kinetic using both the general depletion and modified depletion methods. General depletion was the method that put the tea into solution of a initial F concentration of 0.1 mg/L, 0.2 mg/L, 0.6 mg/L and sampled 2 ml of the solution every 12 h until 84 h. The absorptive quantity was calculated by the difference of F concentration. Modified depletion was the method that put the tea into solution of an initial F concentration of 0.1 mg/L, 0.2 mg/L, 0.4 mg/L, 0.6 mg/L, 0.8 mg/L, 1 mg/L, 2 mg/L and sampled 2 ml of the solution after 24 h. The absorptive rate was calculated by the difference of F concentration. The results on the kinetic parameter by using of the general depletion were different from those by using the modified depletion method. It is because of the initial F concentration. The modified depletion method was a preferable method and the parameters were much reasonable by using of the plotting method and Sigma Plot than the other method. Result showed that the method V_max was 2.605(µg/g*h DW) and 3.419(µg/g*h DW), and K_m was 0.385 mg/L and 0.541 mg/L respectively.

Application of Fertilization Bio-organic System in Tea Plantation of South China

TANG Jin-chi, ZHANG Chi, ZHAO Chao-yi, TANG Hao, LI Jian-long, DAI Jun

Journal of Tea Science. 2008, 28(3): 201-206. doi:10.13305/j.cnki.jts.2008.03.006

Abstract ( 698 )

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Fertilization Bio-organic System (FBO)is applicated in Yingde tea plantation of south China. The tea quality, yield and soil physico-chemical properties were analyzed after one year. Result showed that FBO can obtain the same quality and higher yield under less chemical fertilizer. FBO can improve the physical and chemical characteristics of soil. As a new fertilization management system, FBO will not only promote the soil structure of tea plantation and decrease the fertilization of chemical fertilizer, but also improve soil properties and develop better circulation with the time elapsed. So it has wide application prospects in tea plantation in the future.

Requirement on Nitrogen, Phosphorus and Potassium by Tea Plants with Different Ages under Field Conditions

YOU Xue-qin, YANG Ya-jun, RUAN Jian-yun

Journal of Tea Science. 2008, 28(3): 207-213. doi:10.13305/j.cnki.jts.2008.03.007

Abstract ( 604 )

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In order to study on requirement on nitrogen, phosphorus and potassium by tea plants with different ages under field conditions, the biomass on up-ground parts of Longjing 43 tea plants aged 1, 2, 3, 5 and 10 years in the fall of the first year, and the young shoots, the fallen leaves and the pruning on the second year under different N, P and K application was investigated. The biomass of up-ground parts in tea plants aged 5 years and the concentration of N, P and K in tea plants increased 32, 21, 25, 29 times respectively as compared with those aged one year. The biomass and the concentration of N, P and K in tea plants aged 10 years only increased approximately by 71%, 17%, 13%, 14% respectively as compared with those aged 5 years. The annual requirement of N, P and K for tea plants aged 1~2 years under field conditions is 90.01, 5.71, 31.11 kg/hm² respectively. The annual requirement of N, P and K for tea plants aged 2~3 years is 122.74, 9.63, 45.14 kg/hm² respectively. The annual requirement of N, P and K for tea plants aged 3~4 years is 144.03, 10.33, 47.99 kg/hm² respectively. After pruning, the absorbed N, P and K in part is recovered to the soil through pruning and fallen leaves, the amounts of N, P and K in tea plants used for growth (pruning and fallen leaves are excluded) account for 43.9%, 48.7%, 61.3% of the total respectively. The annual requirement of N, P and K of young tea plants (for example, 5 years aged tea plants) is about 164.4, 12.68, 71.29 kg/hm² respectively. The concentration of N, P and K in the plucking tea plants is mainly used in the growth of tea shoots, and maintain in a stable state and only share a small part of the total (approximately 7.54, 0.53 and 5.44 kg/hm²). A research method on the requirement on N, P and K of tea plants with various growing stage under field conditions was established in this investigation, thus provides a new idea for the further investigation on the nutrient requirement of tea plants.

Characteristics of Soil Nutrients and Enzyme Activity of Ancient Tea Garden in Jingmai, Lancang, Yunnan Province

JIANG Hong, SHA Li-qing

Journal of Tea Science. 2008, 28(3): 214-220. doi:10.13305/j.cnki.jts.2008.03.008

Abstract ( 603 )

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Ancient tea gardens in Yunnan Province are regarded as the provenance of Pu-erh Tea, and the ancient tea garden (ATG) in Jingmai, Lancang County is a typical example of cultivated ATGs. In order to investigate soil nutrients and enzyme activity, the ATG soil in Jingmai and its neighboring conventional tea garden (CTG) and forest soils were sampled. Soil pH, organic matter (OM), total N (TN), total P (TP), total K (TK), alkaline hydrolyzed N (AN), available P (AP), available K (AK), Ca, Al, Mg, activity of acid phosphatase, catalase, urease and saccharase were analyzed. The results showed that (1) All the soils were acid and at a pH of 3.80~3.91, and pH in tea garden soils was lower than in forest soil. (2), The OM, N, P and soil enzyme activity were rich in top soil (0~20 cm) and poor under 20 cm in different soil layers, while K, Mg and Al showed no obvious difference in different layers. (3) OM was rich in all soils, and the contents of OM, TN and AN were CTG < ATG < forest, while TP and AP were richest in ATG. (4) Enzyme activity showed different distribution patterns in the three soils. (5) Soil pH correlated with metal elements, N existed the most significant correlation with other nutrients, and soil nutrients did certain correlation with soil enzymes, especially N, C, P, K.

Comparison on the Different Rehabilitation Methods of Heavy Pruning, Collar Pruning and Replanting

YU Ji-zhong, XU Jia-ming, HUANG Hai-tao, YANG Ya-jun

Journal of Tea Science. 2008, 28(3): 221-227. doi:10.13305/j.cnki.jts.2008.03.009

Abstract ( 665 )

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Replanting, heavy pruning and collar pruning are the main reforming methods for low yield and senescent tea plants. Tea as a perennial crop, it is necessary to evaluate these methods in the long run. A study was conducted in Hangzhou tea experimental station for at least 20 years. The effect of these methods on the yield, production value and economic benefit were compared. The results showed that heavy pruning was a better rehabilitation method for shorter period and more quickly effect, but in the long run, replanting has better economic benefit. It is better to combine replanting and heavy pruning after spring tea to get both short-term and long-term economic benefit in practice.

Synergistic Effect of Tea Saponin on Tripterygium wilfordii EC Against Larvae of Geometrid Ectropis obliqua Prout

DONG Dao-qing, CHEN Jian-ming, YU Xiao-ping, CHEN Lie-zhong, ZHANG Jue-feng

Journal of Tea Science. 2008, 28(3): 228-233. doi:10.13305/j.cnki.jts.2008.03.002

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The synergism of tea saponin on Tirpterygium wilfordii EC against larvae of geometrid Ectropis obliqua Prout was determined. The results indicated that the toxicity of tea saponin to the 2nd instar larvae of E. oblique was not obvious. When treated with leaves that had dipped in tea saponin solution at the concentration of 1,2,4 mg/ml respectively, the corrected mortality was only 27.03% ~ 37.84% on the 7th day after treatment. Toxicity test for T. wilfordii EC against the 2^nd larvae of E. obliqua was studied at the concentration of 0.8, 1.6, 3.2, 6.4 and 12.8 mg/ml respectively. On the 7th day after treatment, the corrected mortality was 27.27%, 33.78%, 40.54%, 56.76% and 63.51% respectively. There were significant synergistic actions on T. wilfordii EC with tea saponin against larvae of E. obliqua. Tea saponin at concentration of 4 mg/ml was added to T. wilfordii EC of 0.8 mg/ml,the corrected mortality of larvae of E. obliqua was significantly increased compared with tea saponin added at the concentration of 1 mg/ml, while no significant differences were found between tea saponin added at the concentration of 2 mg/ml and 4 mg/ml. Treated with T. wilfordii EC mixed with tea saponin of 2 mg/ml, the co-toxicity coefficients for the 2nd instar larvae of E. oblique on 1st, 3rd, 5th, 7th day after treatment, were 667.68, 981.61, 1 706.15 and 1 209.76 respectively. It means that the toxicities increased 5.68~11.10 times, and the amount of T. wilfordii EC decreased over 85% to reach the same effectiveness. The toxicities increased 1.00~6.91 times for the 4th instar larvae of E. obliqua, and the amount of T. wilfordii EC decreased over 50%.

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