Table of Content

25 March 2004 Volume 24 Issue 1

  

Anticancer Activity of Tea: Evidence from Recent Animal Experiments and Human Studies

Hervé Huang, Xin-Qing Xu

Journal of Tea Science. 2004, 24(1):  1-11.  doi:10.13305/j.cnki.jts.2004.01.001

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Hervé Huang, Xin-Qing Xu. Anticancer Activity of Tea: Evidence from Recent Animal Experiments and Human Studies[J]. Journal of Tea Science, 2004, 24(1): 1-11.

   doi:10.13305/j.cnki.jts.2004.01.001

Anticancer activities of tea and tea constituents based on scientific evidences have been established in cellular systems and the underlined molecular mechanisms of action continue to be elucidated in such systems. Several recent papers have thoroughly reviewed available data from these in vitro studies. In this article, we review data from recent studies in experimental animal models, epidemiological studies and human intervention studies/clinical trials.

Advances on DNA Molecular Markers and Gene-engineering in Tea Plants

WANG Li-yuan, CHENG Hao, ZHOU Jian

Journal of Tea Science. 2004, 24(1):  12-17.  doi:10.13305/j.cnki.jts.2004.01.002

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WANG Li-yuan, CHENG Hao, ZHOU Jian. Advances on DNA Molecular Markers and Gene-engineering in Tea Plants[J]. Journal of Tea Science, 2004, 24(1): 12-17.

   doi:10.13305/j.cnki.jts.2004.01.002

Advances on gene-engineering in tea plants are reviewed from following aspects: DNA molecular markers, isolation and cloning of genes, genetic transform system. The problems and probable development in future are also prospected.

Construction of Tender Shoots cDNA Library and Preliminary Analysis of Expressed Sequence Tags Sequencing of Tea Plant

CHEN Liang, ZHAO Li-ping, GAO Qi-kang

Journal of Tea Science. 2004, 24(1):  18-22.  doi:10.13305/j.cnki.jts.2004.01.003

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CHEN Liang, ZHAO Li-ping, GAO Qi-kang. Construction of Tender Shoots cDNA Library and Preliminary Analysis of Expressed Sequence Tags Sequencing of Tea Plant[J]. Journal of Tea Science, 2004, 24(1): 18-22.

   doi:10.13305/j.cnki.jts.2004.01.003

The construction of the first cDNA library of tea plant [Camellia sinensis cv. Longjing 43] in China and the analysis of expressed sequence tags sequencing successful ratio were reported. Total RNA was isolated from tender tea shoots using TRIZOL single-step method, mRNA separated and then the double-strand cDNA amplified by LD-PCR. After size fractionation, the ds-cDNA was ligated to λTripEX2 and recombinant bacteriophages were packaged. The cDNA library was tittered and amplified using XL1-Blue as receptor bacterium. The titer of the original library was 6.8×10⁵pfu/ml with a recombinant rate of 98.05% and 3.5×10⁵ clones in total, the amplified titer was 7.2×10⁹pfu/ml. PCR amplification suggested the inserted cDNA fragments ranged from 0.5 kb to 2 kb, mostly from 1.0 kb to 1.5 kb. The data indicate that the tea cDNA library has high titer, high recombinant percentage and large inserted fragments. Totally, 4320 clones were sequenced, 2963 useful sequences were obtained, corresponding to 68.5%. The first batch of 1687 valid tea plant ESTs were generated.

Study on the Expression of Caffeine Synthase Gene mRNA in Tea Plant

LI Yuan-hua, JIANG Chang-jun, WAN Xiao-chun

Journal of Tea Science. 2004, 24(1):  23-28.  doi:10.13305/j.cnki.jts.2004.01.004

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LI Yuan-hua, JIANG Chang-jun, WAN Xiao-chun. Study on the Expression of Caffeine Synthase Gene mRNA in Tea Plant[J]. Journal of Tea Science, 2004, 24(1): 23-28.

   doi:10.13305/j.cnki.jts.2004.01.004

The expression of caffeine synthase gene mRNA in tea plant by means of in situ hybridization was investigated. Results showed that the expressional sites is mainly located in the cell cytoplasm and nucleus of palisade tissue, but not in vacuole. The expressional sites of leaf vein are mainly in parenchyma and phloem, however, that in phloem is weaker. The expressional sites in young stalk are in phloem. The expressional sites in different cultivars are basically similar, but the expressional signal is different. Results also showed that the expressional signal of the first fresh leaf in spring is stronger than that in autumn. In terms of processing technique, the green teas which are spread 0.5 h、1 h、2 h showed apparent expressional signal, but the signal was not observed in the black teas withered for 6 h、8 h、10 h and Oolong teas shaken for 6 h、8 h、10 h.

Breeding Report on Strong Aroma and Good Quality Newly Bred Oolong Variety Rui Xiang

CHEN Rong-bing, HUANG Fu-ping, CHEN Chang-song, YOU Xiao-mei

Journal of Tea Science. 2004, 24(1):  29-32.  doi:10.13305/j.cnki.jts.2004.01.005

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CHEN Rong-bing, HUANG Fu-ping, CHEN Chang-song, YOU Xiao-mei. Breeding Report on Strong Aroma and Good Quality Newly Bred Oolong Variety Rui Xiang[J]. Journal of Tea Science, 2004, 24(1): 29-32.

   doi:10.13305/j.cnki.jts.2004.01.005

Rui Xiang was selected from progenies of natural hybrid of Huang Dan cultivar. It is a newly bred Oolong tea cultivar with strong aroma and good quality obtained through systematic selection. Its plucking begins late in April, similar to that of Tie Guanying. The average yields in three tests were above 10% higher than that of Huangdan. Obtained Oolong tea quality is better than Huangdan with heavier appearance, strong fragrance and a better taste as well. Total water soluble, polyphenols, caffeine, catechins, EGCG and free amino acids were 43.4%, 24.1%, 2.9%, 14.6%, 5.4% and 3.0%, respectively. The total aroma oil determined in its made tea was 128.98, which was greater than Huangdan. Its main aroma components were nerolidol, benzaldehyde, jasmine jactone, α-farnesene, β-ionone and cis-jasmone. Its adaptation to environment and resistance to adverse condition was similar to Huangdan.

Analysis of Genetic Relationships Among “Rucheng Baimao Cha” Plants with RAPD Method

LI Xing-hui, LIU Chun-lin, SHI Zhao-peng, LUO Jun-wu, SHEN Cheng-wen, GONG-zhihua, CHEN-xuan

Journal of Tea Science. 2004, 24(1):  33-36.  doi:10.13305/j.cnki.jts.2004.01.006

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LI Xing-hui, LIU Chun-lin, SHI Zhao-peng, LUO Jun-wu, SHEN Cheng-wen, GONG-zhihua, CHEN-xuan. Analysis of Genetic Relationships Among “Rucheng Baimao Cha” Plants with RAPD Method[J]. Journal of Tea Science, 2004, 24(1): 33-36.

   doi:10.13305/j.cnki.jts.2004.01.006

The genetic relationships of the individual plants among the “Rucheng Baimao Cha” population were studied with the RAPD method. The results were as follows: The maximum number of PCR bands of the individual plants among the “Rucheng Baimao Cha” population was 13,and the minimum number was 10. The maximum DNA polymorphism amplified by RAPD-PCR was 92.31%, while the minimum was 70.00%; the average was 85.40%. This result showed the abundant genetic diversities. The maximum genetic distances among individual plants of the “Rucheng Baimao Cha” population was 0.5791; the minimum one was 0.0882. The result from the DNA molecular dendrogram showed that the variation among individual plants of“Rucheng Baimao Cha” population was continuous. The cluster analysis by UPGMA showed that the “Rucheng Baimao Cha”population could be divided into 6 groups. This result matched the morphologic characteristics of the population. From the DNA molecular dendrogram it was showed that the key germplasm groups were protected in the Rucheng Baimao Cha Demonstration Farm in Rucheng county, Hunan province, P. R. China.

Sporogenesis and Gametogenesis of Camellia ptilophylla

AO Cheng-qi

Journal of Tea Science. 2004, 24(1):  37-40.  doi:10.13305/j.cnki.jts.2004.01.007

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AO Cheng-qi. Sporogenesis and Gametogenesis of Camellia ptilophylla[J]. Journal of Tea Science, 2004, 24(1): 37-40.

   doi:10.13305/j.cnki.jts.2004.01.007

To Camellia ptilophylla, the cytokinesis at the time of pollen mother cell meiosis conforms to simultaneous type. The arrangment of microspores in tetrad is isobilateral.The mature pollen grain is of the 2-cell type.The development of the anther wall follows Basic-type,tapetum glandular,ovules anatropous, bitegminous, tenuinucellate and the embryo sac belongs to Allium type.

Cloning of Aluminum-tolerant Associated Gene from Fungus ALF-1(Neurospora sp.) in Rhizospheric Soil of Tea Plant

LU Jian-liang, LIANG Yue-rong, WU Ying, HUANG Fu-ping

Journal of Tea Science. 2004, 24(1):  41-43.  doi:10.13305/j.cnki.jts.2004.01.008

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LU Jian-liang, LIANG Yue-rong, WU Ying, HUANG Fu-ping. Cloning of Aluminum-tolerant Associated Gene from Fungus ALF-1(Neurospora sp.) in Rhizospheric Soil of Tea Plant[J]. Journal of Tea Science, 2004, 24(1): 41-43.

   doi:10.13305/j.cnki.jts.2004.01.008

Aluminum-tolerant associated gene of fungus ALF-1(Neurospora sp.) in rhizospheric soil of tea plant was cloned using rt-PCR technique. The gene expression was induced by aluminum treatment, and its sequence was 359 base pair (bp). The opening reading frame (ORF) was 260bp which situated from 100^th to 359^thbp, and encoded 86 amino acid residues. BLAST analysis showed that the gene sequence obtained was 50% identity to that of Arabidopsis thaliana (thale cress).

Pharmacokinetics and Bioavailability of Catechin Liposome in Rabbits

SUN Zhi-liang, Liu Xiang-xin, Ling Qin-lu, Liu Zi-kui, Liu jin-fei, He Xiang rong

Journal of Tea Science. 2004, 24(1):  44-48.  doi:10.13305/j.cnki.jts.2004.01.009

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SUN Zhi-liang, Liu Xiang-xin, Ling Qin-lu, Liu Zi-kui, Liu jin-fei, He Xiang rong. Pharmacokinetics and Bioavailability of Catechin Liposome in Rabbits[J]. Journal of Tea Science, 2004, 24(1): 44-48.

   doi:10.13305/j.cnki.jts.2004.01.009

Pharmacokinetics and bioavailability of Catechin Liposome were studied in 10 healthy rabbits following single intravenous (25mg/kg) and oral administration (25mg/kg)of the drug. Plasma concentrations of. Catechin Liposome were determined by high-performance liquid chromatography. The concentration-time data were fitted to a two-compartment model following a single intravenous injection in rabbits. The main pharmacokinetic parameters were as follows: distribution half-life (t_1/2α) (0.18±0.01)h,elimination half-life (t_1/2β) (1.52±0.08)h,apparent distribution volume (Vd) (4.48±0.24)L total body clearance (Cl_B) (2.05±0.07)L/h,the area under curve (AUC) (29.20±1.00)mg/(L.h). The drug concentration-time data also were fitted to a one-compartment open model with first order absorption after a single oral administration Catechin Liposome. The pharmacokinetic parameters were as follows: t_1/2k_α (0.27±0.03)h, t_1/2k_e(1.72±0.04)h, t_max (0.87±0.05) h, C_max 6.53±0.62βmg/L, AUC 25.90±1.34βmg/(L.h), bioavailability F 88.60±5.73%. Pharmacokinetic characteristics of Catechin Liposome in healthy rabbits showed a rapid absorption, the time to reach Cmax is short, slow elimination, long half-life, large apparent distribution volume, and high bioavailability of oral administration. The results showed：after Catechin was prepared in liposome form, the tissue distribution and pharmacokinetics were changed significantly.

Study on Lipid-depressing Function of Kuding Tea Prepared from Ilex latifolia Thunb Leaves

PAN Hui-juan, LIAO Zhi-yin, YING Qi-cai, TIAN Ju-wen, XU Ying

Journal of Tea Science. 2004, 24(1):  49-52.  doi:10.13305/j.cnki.jts.2004.01.010

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PAN Hui-juan, LIAO Zhi-yin, YING Qi-cai, TIAN Ju-wen, XU Ying. Study on Lipid-depressing Function of Kuding Tea Prepared from Ilex latifolia Thunb Leaves[J]. Journal of Tea Science, 2004, 24(1): 49-52.

   doi:10.13305/j.cnki.jts.2004.01.010

The hyperlipidemia rats were fed with different dose of distillable liquid of Ilex latifolia Thunb leaves. The Transansio automatic biochemical analyzer was used to determine the serum lipid rats in normal and model groups . The results showed that the Ilex latifolia Thunb tea and yuejiancao oil have significantly effects in decreasing the serum triglyceride and total cholesterol in the hperlipidemia rats(p<0.05).It can also modulate the HDL-C 、LDL-C 、AI and R-CHR indexes. The resuls implicated that the Ilex latifolia Thunb has significantly effects in decreasing the blood lipid of hyperlipidemia and in preventing atherosclerosis.

Studies on Preparing Theaflavins from Oxidation of Tea Catechins in Vitro

WANG Kun-bo, LIU Zhong-hua, HUANG Jian-an

Journal of Tea Science. 2004, 24(1):  53-59.  doi:10.13305/j.cnki.jts.2004.01.011

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WANG Kun-bo, LIU Zhong-hua, HUANG Jian-an. Studies on Preparing Theaflavins from Oxidation of Tea Catechins in Vitro[J]. Journal of Tea Science, 2004, 24(1): 53-59.

   doi:10.13305/j.cnki.jts.2004.01.011

Preparaing theaflavins from the oxidation of tea catechins in vitro simulated fermentation were studies. Two ways of preparing theaflavins was compared. Results of enzymatic oxidation showed that the highest content of theaflavins (15.05%) was formed at 80 minutes fermentation with polyphenol oxidase. Results of chemical oxidation also showed that the optimum condition for the formation of theaflavins is using the catechins from large-leaf tea plant on the raw material with concentration of 10βmg/ml and the ratio of oxidants in 2:3:1 (catechins:K₃Fe(CN)₆:NaHCO₃).

Analysis of the Xanthophyll Cycle Component in Fresh Leaves of Tea Plant by High Performance Liquid Chromatography and Its Photo-protection

WEI Chao-ling, JIANG Chang-jun, TAO Han-zhi, HU Ying-hui

Journal of Tea Science. 2004, 24(1):  60-64.  doi:10.13305/j.cnki.jts.2004.01.012

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WEI Chao-ling, JIANG Chang-jun, TAO Han-zhi, HU Ying-hui. Analysis of the Xanthophyll Cycle Component in Fresh Leaves of Tea Plant by High Performance Liquid Chromatography and Its Photo-protection[J]. Journal of Tea Science, 2004, 24(1): 60-64.

   doi:10.13305/j.cnki.jts.2004.01.012

The xanthophyll cycle components in fresh leaves of tea plant (Camellia sinensis cv. Fudingdabai)were extracted by acetone. The pigments were analyzed with a Spherisorb column （C₁₈ , 5μm,250㎜×4.6βmm）,and were eluted using acetonitrile: methanol(85:15) and methanol:ethyl acetate(68:32) by a linear gradient. The pigments were detected by their absorbance at 445βnm. It was showed that violaxanthin, antheraxanthin and zeaxanthin from fresh leaves of tea plant were detected by HPLC, respectively, under temperatures of 15℃, 22℃ and 30℃, meanwhile, the pigments were separated well, and it is a sensitive, well-reappeared method. Tea plant had a larger xanthophyll cycle pool and a larger de-epoxidation state in higher sun-light, which indicated xanthophyll cycle plays an important role in photo-protection in tea plant.

Effect of Cadmium Ions on Photosynthetic Structure and Its Functions of Tea Leaves

SU Jin-wei, WANG Xiang-Ping

Journal of Tea Science. 2004, 24(1):  65-69.  doi:10.13305/j.cnki.jts.2004.01.013

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SU Jin-wei, WANG Xiang-Ping. Effect of Cadmium Ions on Photosynthetic Structure and Its Functions of Tea Leaves[J]. Journal of Tea Science, 2004, 24(1): 65-69.

   doi:10.13305/j.cnki.jts.2004.01.013

Tea plant was cultivated in Hoagland nutrient solution containing 0.5βmmol/L Cd²⁺ for 8 days. Tea leaves lost their green color gradually with the change in the ultrastructure of the chloroplast . At the early stage, there was a slight decrease in the number of granum thylakoids. Then, the thylakoids piled up loosely, and chloroplasts rounded off to different extents. At the late stage , the structure of the chloroplast related to thylakoid became unclear, while the thylakoid system was disintegrated abnormally with a few single bent and swell thylakoids, and some ruptured envelope of chloroplast. Cadmium ions significantly inhibited the photosynthesis activity and led to the decline in the chlorophyll content and chlorophyll a/b ratio. Further more the photochemical efficiency of PSII（Fv/Fm）and the potential activity of PSII（Fv/Fo）decreased in different degrees with increasing Cd²⁺ stress . The pollution duration resulted in the damage of photosynthetic structure in tea leaf cells.

Effects of Zuoqing Intensity on the Protein Composition, Polyphenol Oxidase and Esterase Isozymogram of the Fresh Leaves during Oolong Tea Manufacture

HUANG Fu-ping, LIANG Yue-rong, CHEN Rong-bin, CHEN Wei, KE Yu-qin, LU Jian-lian

Journal of Tea Science. 2004, 24(1):  70-74.  doi:10.13305/j.cnki.jts.2004.01.014

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HUANG Fu-ping, LIANG Yue-rong, CHEN Rong-bin, CHEN Wei, KE Yu-qin, LU Jian-lian. Effects of Zuoqing Intensity on the Protein Composition, Polyphenol Oxidase and Esterase Isozymogram of the Fresh Leaves during Oolong Tea Manufacture[J]. Journal of Tea Science, 2004, 24(1): 70-74.

   doi:10.13305/j.cnki.jts.2004.01.014

Using Rougui (Camellia Sinensis (L.) var.) fresh leaves as material, fresh leaves soluble protein content and its composition by SDS-PAGE, polyphenols oxidase activity and its isozymogram,and esterase isozymogram were investigated during Oolong tea Zuoqing procedure with different Zuoqing intensity. The results indicated that, during zuoqing process, the soluble protein content and its protein components decreased, and decreased sharply with the intensity, during the periods polypenol oxidase activity increased at the earlier stage, and decreased at the later, its isozymogram checked out 7 bands, no new bands was found during the period, and its E2 band activity apparently enhanced while its E6 activity decreased. Esterase had 7 isozymogram bands, the changes of the each band activities was not remarkable in the whole procedure. The relations of the factors mentioned above to the Oolong quality formation also were discussed in the paper.