Journal of Tea Science

Genetic Diversity of Ancient Tea Plant in Yunnan Province of China Revealed by Inter-simple Sequence Repeat(ISSR) Polymerase Chain Reaction

JI Peng-zhang, ZHANG Jun, WANG Ping-sheng, HUANG Xing-qi, XU Mei, TANG Yi-chun, LIANG Ming-zhi

Journal of Tea Science. 2007, 27(4): 271-279. doi:10.13305/j.cnki.jts.2007.04.001

Abstract ( 553 )

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Tea plant [Camellia sinensis var. assamica] populations in Yunnan province of China cultivated since ancient times are important genetic resources for future breeding of new tea cultivars. To evaluate the genetic diversity within and between Yunnan ten representative populations and to formulate conservation strategies for the ancient tea populations in Yunnan was conducted by ISSR technique in this study. The genetic diversity within populations calculated from Nei's genetic diversity (He) was estimated to be 0.281, on average, at the population level and 0.461 at the variety level, while Shannon indices (Ho) of population level and Assam variety level were 0.418 and 0.653, respectively and the percentage of polymorphic loci (P) of the ten populations ranged between 56.5% to 90.9%, respectively. Coefficients of gene differentiation (Gst) within the population was 0.391, in agreement with the result of Shannon’ diversity indices analysis (36.0%) and analysis of molecular variance (AMOVA) among population (39.7%), which illuminated that 60.9% of the variation in Assam variety were from the individuals of the population, and 39.1% of the genetic variation were from the population. The result indicated Camellia var. assamica variety possesses high genetic diversity and medium genetic differentiation was existed in the genetic variation between population. This could be caused by the highly outcrossing nature and habitat fragmentation of tea species. Conservation strategy are suggested including in situ strategies and ex situ strategies based on the observed genetic information of population.

AFLP Analysis on Genetic Diversity of Fenghuang-Dancong Tea Plant Germplasms

XIAO Li-zheng, YAN Chang-yu, LI Jia-xian, LUO Jun-wu, HE Yu-mei, ZHAO Chao-yi

Journal of Tea Science. 2007, 27(4): 280-285. doi:10.13305/j.cnki.jts.2007.04.002

Abstract ( 421 )

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Genetic diversity on 34 Fenghuang-Dancong tea plants were evaluated by AFLP fingerprinting using 5 primer combinations. The 5 primer combinations generated 438 bands and 87.6 bands were amplified by each primer pair on average, 348 bands(79.3%) polymorphic AFLP markers were detected, The result indicated that the genetic distance(GD) based on AFLP data by using DPS2000 was significant. The varience range of GD among the 34 Fenghuang-Dancong was 0.13~0.49. The GD(0.49) is highest between Zimaohuangzhixiang and Funanmilan and between Baiyedancong and Tongtianxiang, however, the GD(0.13) between Da-ansongcha and Zongsuojia is low. The dendrogram indicated that the genetic of 34 Fenghuang-Dancong germplasms have no certain relation with the aroma type.

Preliminary Study on Differential Gene Expression During Cold Acclimation in Tea Plant (Camellia sinensis)

MEI Ju-fen, WANG Xin-chao, YANG Ya-jun, LI Xing-hui

Journal of Tea Science. 2007, 27(4): 286-292. doi:10.13305/j.cnki.jts.2007.04.003

Abstract ( 495 )

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Differential gene expression during cold acclimation was studied by mRNA differential display reverse transcription-PCR (DDRT-PCR) in tea plant (Camellia sinensis). Twenty-three differentially expressed cDNA fragments were steadily re-amplified. Five fragments were confirmed by RT-PCR analysis. Among them, three were up-regulated while other two were down-regulated during the cold acclimation process. Sequencing of three differential fragments revealed homology to putative cis-zeatin-O-glucosyl-transferase of Sorghum （Sorghum bicolor）, chloroplast gene for chloroplast product of cucumber(Cucumis sativus),ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit (rbcS) mRNA of tea plant(Camellia sinensis), respectively, while the other two showed no homologous to known genes.

Effect of SOD on Anti-cancer Activity and Stability of Epigallocatechin-3-gallate

ZHANG Bin, XIA Yan-qiu, JIN Li-ji, XU Yong-ping

Journal of Tea Science. 2007, 27(4): 293-298. doi:10.13305/j.cnki.jts.2007.04.006

Abstract ( 410 )

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Effects of epigallocatechin-3-gallate (EGCG) on the growth of human colonic cancer cells HT-29, human breast cancer MCF-7 and human cervical cancer Hela were investigated by MTT assay. Cytotoxicity of EGCG on these cells was detected by determining the activity of LDH in culture medium. Induction of apoptosis in HT-29 cell after the EGCG treatment was analyzed by flow cytometry. Stability of EGCG in DMEM/F-12 was determined by HPLC and all studies above were compared in the presence or absence of SOD. Investigation showed that cells survival percentage deceased and activity of LDH in the culture medium increased after adding SOD. The apoptotic rate of HT-29 and the stability of EGCG in culture increased in the presence of SOD comparing to absence of SOD.

Study of Green Tea Polysaccharides on Lipid Peroxidation of Plasma Lipoproteins

YIN Xue-zhe, XU Hui-xian, JIN Hua

Journal of Tea Science. 2007, 27(4): 299-301. doi:10.13305/j.cnki.jts.2007.04.004

Abstract ( 434 )

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The oxygen free radical scavenging activities and anti-lipoprotein oxidative effects of green tea polysaccharides were studied. ROO·, O₂^-· and OH· scavenging activities were studied with chemiluminescence analysis, H₂O₂ scavenging activity was determined by spectrophotometry. Very low density lipoprotein (VLDL), low density lipoprotein (LDL) and high density lipoprotein (HDL) were isolated from fasting plasma by sequential ultracentrifugation at 0βh and 2βh after 20 healthy volunteers ingested green tea polysaccharides and degrees of peroxidization and susceptibilities of lipoproteins to oxidation were investigated. The results showed that green tea polysaccharides exhibited scavenging effects on ROO·, O₂^-·, OH· and H₂O₂ in vitro, significantly decreased lipid peroxides of plasma, VLDL, LDL and HDL; noticeably prolonged lag phage of LDL oxidation curve, indicating reduced susceptibility of LDL to oxidation. It is suggested that green tea polysaccharides can increase antioxidant capacities of plasma and lipoproteins, therefore may show the function of preventing cardiovascular disorders and delaying ageing.

(-)-epigallocatechin Gallate (EGCG) Enhance the Anticancer Activity Induced by Carmustine in vitro

HUANG Xiu-hua, ZHANG Dan, HAO Jin, YANG Zu-jun, REN Zheng-long

Journal of Tea Science. 2007, 27(4): 302-306. doi:10.13305/j.cnki.jts.2007.04.007

Abstract ( 483 )

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The effect of (-)-epigallocatechin gallate (EGCG) combined with carmustine on the growth inhibition and DNA damage of human lung carcinoma cell line A549 were evaluated by MTT assay and Comet assay repectively. The results showed that 50% inhibitory concentration (IC₅₀) of carmustine to A549 human lung carcinoma cell line was decreased when combined with EGCG. The 50% inhibitory concentration (IC₅₀) of carmustine alone to human lung carcinoma cell line A549 was 187.46βμg/ml, When the carmustine is combine with EGCG(25βμg/ml and 50βμg/ml, the concentration under which EGCG has no growth inhibition to A549), however, the IC₅₀ value to A549 cell line decreased to 139.56βμg/ml and 154.02βμg/ml respectively. EGCG also enhanced carmustine-induced DNA damage in A549 cell line by Comet assay. The tail moment increased from 19.02±14.60 to 33.07±10.47. So, EGCG can enhance carmustine-induced DNA damage in A549 cell line, hence enhance the anticancer activity of carmustine.

Study on the Toxicity Experiments of Fuzhuan Tea

XIAO Wen-jun, FU Dong-he, REN Guo-pu, GONG Zhi-hua, XIAO Li-zheng, LIU Zhong-hua

Journal of Tea Science. 2007, 27(4): 307-310. doi:10.13305/j.cnki.jts.2007.04.008

Abstract ( 557 )

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The toxic experiment of Fuzhuan tea was carried out by using the extracted instant Fuzhuan tea as material in order to evaluate its safety for the mice and rat. The results showed that the LD₅₀ of male mice and female mice was 14β700βmg/kg·bw and 19β600βmg/kg·bw respectively(the limit reliability was 10β600~36β000), and both of the Ames test and the semen teratogenicity results were negative. During the 30 days fed by the feeding mixed with 0.85%, 1.25%, 1.70% instant Fuzhuan Tea for the rat rats of each dose group grew well, the hemogram indexes of rats from each dosage, blood biochemical indexes and triglyceride, Urea anhydride, blood muscle anhydride and blood sugar indexes, ratio of spleen to body weight, kidney to body weight, and the histopathological characteristic of all test rats showed no significantly difference (P<0.05), which suggested Fuzhuan Tea was a no poisonous food.

Study on the Process of Machine-processed Needle-Shaped Famous Tea

YU Zhi, YANG Yan, ZHOU Ji-rong, NI De-jiang

Journal of Tea Science. 2007, 27(4): 311-315. doi:10.13305/j.cnki.jts.2007.04.009

Abstract ( 444 )

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Orthogonal regression method was adopted to optimize the process of needle tea by 6CZZ-600 tea shape machine. The result showed that shaping temperature had a significantly effect on the content of tea polyphenol and chlorophyll, and leaf weight had a significantly effect on amino acid and chlorophyll, but the turning rate had little effected on them. The content of amino acid and soluble sugar increased first, and then declined according to the rising temperature, the content of tea polyphenol and chlorophyll declined all along. Too high temperature accelerated the reducing of chlorophyll, but shaping in low temperature would lead to a boring aroma. The content of tea polyphenol and chlorophyll declined according to the increasing of leaf weight, but the content of amino acid and soluble sugar increased first, and then declined. Too much leaf would lead to bad color of green tea, but too little could not achieve good quality of tightness and thinness. Combination the demand of excellent needle tea quality and the efficiency of producing, the optimized mechanical process of excellent needle green tea was: leaf weight was 100βg to 150βg per trough, at 90℃ to 110℃, rotate rate was 184βr/min to 192βr/min.

Influence of the Scenting Technique Factors on Quality of Chimonanthus Tea

PANG Xiao-li, SI Hui-qing, LI Yong-ju

Journal of Tea Science. 2007, 27(4): 316-322. doi:10.13305/j.cnki.jts.2007.04.010

Abstract ( 380 )

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For the purpose of optimizing the technological parameter for chimoncanthus tea scenting technique, the influence on quality of chimonanthus tea for the single scenting technique factors by means of Two Regression Orthogonal Combination Test was investigated. Results showed that the technique factors affected obviously on organoleptic quality of the chimonanthus tea, but not influenced on their bio-chemical composition significantly. The water content of base tea is the most remarkable among all the factors in influencing the quality of chimonathus tea, then, in proper order, the chimonanthus flower weight percentage(%), the scenting time, and the stack temperature. The optimum combination of technological parameters is: water content 10.60%, flower weight percentage 42.20%, temperature 11.3℃, and time 32.4βh.

Positive Studies on Characteristics of Tea Consumers in Hangzhou City

TANG Yi, HUANG Han-dan

Journal of Tea Science. 2007, 27(4): 323-327. doi:10.13305/j.cnki.jts.2007.04.011

Abstract ( 459 )

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According to the results of questionnaire survey in Hangzhou city, Ordinal Logit model was applicated to explain relations between characteristics of consumers and level of tea demand. The regression results showed: custom of tea drinking showed the most important impact on tea consumption amount of Hangzhou citizen, moreover the major factor to affect purchasing price of tea is level of income. Other relevant factors include age, degree of trust in tea’s efficacy, price sensitivity and so on, whereas gender, educational background and occupation showed no obvious effect on tea demand.

Study on the Method of Fetching Unbroken Tea Color with Computer

LI Jie, QI Gui-nian

Journal of Tea Science. 2007, 27(4): 328-334. doi:10.13305/j.cnki.jts.2007.04.012

Abstract ( 426 )

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A new method of fetching unbroken tea color with computer,named L Method,has been established. The accuracy of the method was verified,and the further investigation also has been fulfilled. The results showed that the L method can fetch the color of tea accurately and quickly. Scanned with resolution 100βdpi,the credible interval(95%) of the result is: L=0,-0.45≤a≤0.45,-0.71≤b≤0.71. When the scanned area is 11βcm×11βcm or more,the color parameters of tea scanning photo may fully stand for the tea actual color. The SE is: -0.16≤L≤0.16,-0.13≤a≤0.13,b=0. There was obvious distinction between the color of unbroken tea and crush tea, so the later can't represent the tea actual color. Using computer to fetch color parameters is better than chroma meter.

A Preliminary Attempt to Establish the Digital Multiple Chemical Fingerprints of Green Tea

WANG Li-yuan, CHENG Hao, ZHOU Jian, YE Yang, LIU Xu, LU Wen-yuan

Journal of Tea Science. 2007, 27(4): 335-342. doi:10.13305/j.cnki.jts.2007.04.013

Abstract ( 491 )

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The digital multiple chemical fingerprints of nine groups of flat-shaped green tea produced in different district of Hangzhou, Lishui and Xinchang city, and made from three cultivars, including Longjing 43, jat and Yingshuang, were made by conjoining of two HPLC chromatograms, after the sample preparation procedure being optimized for better extraction and less isomerization. Also validation analysis was made by using 14 tea samples collected independently from above mentioned districts, and similarities between those 9 kinds of pattern fingerprints were analyzed. The result showed that those established pattern fingerprints well represented the characteristic attributes of those origin samples respectively. Distinct differences were found between the pattern fingerprints of samples from different district using same cultivar, or from different cultivar made in same district, but the difference between the former was not obviously as much as that between the latter. So it was reminded that premier green teas should be made from one optimum cultivar or at least several similar suitable cultivars to guarantee the unification of the flavor and the character.

Study on HPLC Method for the Analysis of Trace Pigments in Ready-to-drink Tea

PAN Shun-shun, LU Jian-liang, YANG Xiao-li, ZHENG Xin-qiang, DU Ying-ying, Devijat B, LIANG Yue-rong*

Journal of Tea Science. 2007, 27(4): 343-348. doi:10.13305/j.cnki.jts.2007.04.014

Abstract ( 376 )

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Study on HPLC method for analyzing of trace pigments in ready-to-drink tea was conducted by using different eluants with acetonitrile as mobile phase. Result showed that gradient elution was suitable for analyzing by using acetonitrile/acetic acid/water in ratio of 3/0.5/96.5(v/v/v) as mobile phase A, and using acetonitrile /methanol /chloroform in ratio of 75/15/10(v/v/v) as mobile phase B. The optimized elution time program could be described that phase B increased gradiently from 80% to 100% in early 20 minutes, then remained 100% for 15 minutes. The main pigments, i.e. neoxanthin, violaxanthin, lutein, chlorophyll b, chlorophyll a, phaeophytin b, β-Carotene and phaeophytin a, could be separated upto baseline under that condition. Verification test showed that the optimized HPLC method with good repeatability and low detection limits was suitable for measurement of trace pigments in ready-to-drink tea.

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