Journal of Tea Science

Genetic Diversity of Camellia taliensis from Yunnan Province of China Revealed by AFLP Analysis

JI Peng-zhang, WANG Yun-gang, JIANG Hui-bing, TANG Yi-chun, WANG Ping-sheng, ZHANG Jun, HUANG Xing-qi

Journal of Tea Science. 2009, 29(5): 329-335. doi:10.13305/j.cnki.jts.2009.5.001

Abstract ( 534 )

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Camellia taliensis is a critically species endemic to southern Yunnan, China. We assessed the genetic variability within and among eleven populations of this species using AFLP marker. At species level: Nei’s (1973) gene diversity (He) was 0.099, and Shannon’s Information index (Ho) 0.178, at the population level: He=0.083, Ho =0.137. A low level of genetic differentiation among populations was detected based on Gst =0.1606 (16.1%), Shannon’s diversity index (16.04%), and AMOVA (19.03%). Pairwise genetic identity (I) values among populations ranged from 0.971 to 0.997. There was no correlation between genetic and geographic distance among the populations studied. The influence of human activity and forest fragmentation may play a prominent role in creating this species’s current endangered status. Conservation strategies are suggested including in situ strategies and ex situ strategies based on the observed genetic information of population.

Prokaryotic Expression of α-tubulin Gene of Camellia sinensis and Preparation of α-tubulin Polyclonal Antibody

TAN Zhen, TONG Xin, FANG Chao, JIANG Chang-jun, CHEN Cong

Journal of Tea Science. 2009, 29(5): 336-340. doi:10.13305/j.cnki.jts.2009.5.002

Abstract ( 505 )

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According to the relevant complete mRNA sequence of α-tubulin gene of Camellia sinensis (accession NO. DQ340766), the coding region of α-tubulin gene was amplified using RT-PCR method, and its products were expressed in Escherichia coli using the pET-32a(+) vector. The fusion protein was expressed in the form of inclusion body when induced with isopropyl-D-thiogalatopyranoside (IPTG), then was purified to immunize the rabbit to produce the polyclonal antibody against α-tubulin. The specificity of the antibody was confirmed by Western blot analysis. The results of Western blot revealed well specificity of the antibody against α-tubulin in Camellia sinensis plant.

Phylogenetic Analysis of Interspecies in Section Thea Through SSR Markers

WANG Li-yuan, LIU Ben-ying, JIANG Yan-hua, DUAN Yun-shang, CHENG Hao, ZHOU Jian, TANG Yi-chun

Journal of Tea Science. 2009, 29(5): 341-346. doi:10.13305/j.cnki.jts.2009.5.003

Abstract ( 497 )

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For the purpose of investigating the transferability and polymorphism information content (PIC) of different types of SSR primers, PCR amplification for 41 accessions from section thea was conducted in this investigation. The results showed that the SSR primers form Ser. Sinensis were highly transferable among section Thea. The polymorphic and transferable primers account for 60% of EST-SSR markers. Then the genetic diversity among 12 species of section Thea was evaluated through SSR analysis. The number of alleles in different loci ranged from 2 to 6, with average 4.21 per locus. The phyolgenetic dendrogram was constructed by the UPGMA.

Molecular Cloning and Real-Time PCR Analysis of Flavone SynthaseⅡ Gene Full-length cDNA from the Tea Plant

QIAO Xiao-yan, MA Chun-lei, CHEN Liang

Journal of Tea Science. 2009, 29(5): 347-354. doi:10.13305/j.cnki.jts.2009.5.004

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Using RT-PCR and RACE techniques, flavone synthase Ⅱ (FSⅡ) gene, the key enzyme in synthesize flavone, was cloned in the tea plant. The FS II belongs to cytochrome P450 superfamily. The GenBank Accession No. is FJ169499 in the NCBI. The FSⅡ gene cDNA had 1 824 bp in full-length with 1 605 bp in coding region, encoding 534 amino acids. The putative molecular weight was 60.4 kD. The expression levels of flavone synthaseⅡ gene in spring ‘two and a bud’, mature leaves, petals, stamen and pistils, and seeds of the tea plant were assessed using SYBR Green I Real-time PCR. The expression in mature leaves was significantly higher than that in the other four tissues.

ISSR Markers for Discriminating Tea Germplasm Resources from Yunnan Province

LIU Ben-ying, WANG Li-yuan, LI You-yong, TANG Yi-chun, HE Wei, CHENG Hao, WANG Ping-sheng

Journal of Tea Science. 2009, 29(5): 355-364. doi:10.13305/j.cnki.jts.2009.5.005

Abstract ( 458 )

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It was of much importance using minimum primers to obtain the maximum identification ability．Molecular identification of 134 tea germplasm resources using inter-simple sequence repeat (ISSR) markers was conducted in this study. There were 3 independent ways to identify tea germplasms: a) unique ISSR markers;b) unique band patterns and c) a combination of the band patterns provided by different primers．The result showed that the presence of 10 unique ISSR markers and the absence of 15 unique markers obtained from 12 primers made it possible to identify 21 tea germplasms, including ‘Xiangzhuqingdashancha’ and so on. Using 54 unique band patterns of primer UBC811 could identify 35 tea germplasms, including ‘Hainandayecha 1’ and so on. A1l the 134 tea germplasms could be entirely identified by the band patterns combination of primer UBC811, UBC835, ISSR2 and ISSR3, which was successfully used to construct the ISSR fingerprinting for discriminating 134 tea germplasms from Yunnan Province.

Study on the Changes of Non-galloylated Catechins and Relative Enzymes in Tea Shoots

ZHANG Xian-lin, GAO Li-ping, XIA Tao, LIU Ya-jun, GAO Ke-jun

Journal of Tea Science. 2009, 29(5): 365-371. doi:10.13305/j.cnki.jts.2009.5.006

Abstract ( 523 )

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The changes of non-galloylated catechins and relative enzymes as well as the composition of catechins in different parts of tea shoots and in different seasons were investigated in this paper. The results showed that the contents of non-galloylated catechins except GC increased gradually from bud to the fourth leaf. Both of two synthetical enzymes related to non-galloylated cahechins, the activity of DFR/LAR declined gradually, but the activity of ANR was the highest in the third leaf. The result of correlation analysis showed that the contents of total catechins in different tea leaves presented a prominent positive correlation with the activity of DFR/LAR,but not with the activity of ANR.

Taste Characteristics of Purple Tea Leaf and the Reduction of Bitterness and Astringency

ZHAO Xian-ming, WANG Xiao-shi, DU Xiao

Journal of Tea Science. 2009, 29(5): 372-378. doi:10.13305/j.cnki.jts.2009.5.007

Abstract ( 500 )

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The quality of the green tea made from purple tea leaf, which is common in tea production, is not so good for its evident bitterness and astringency. In this paper, the main bitterness and astringency characteristic components of purple tea leaf were separated and determined, and the taste characteristics of those components were quantitatively evaluated by the threshold and index of bitterness and astringency. The results showed that the content of anthocyanidins was as high as 1.14%, and the contents of anthocyanidins and catechins of purple tea leaf were 57 and 1.13 times to those of green tea leaf, their higher content was mainly responsible to the bitterness and astringency of purple tea leaves. The order of threshold of those components was caffeine (0.30) < anthocyanidins (0.40) < catechins (0.80), and the order of bitterness and astringency index was catechins (3.32) > caffeine (2.13) > anthocyanidins (0.57), which was also used to evaluate the effect of components on the taste of tea. Furthermore, adding 0.2% casein into purple tea leaf during green tea processing could reduce the bitterness and astringency, the quality could be promoted, the concentration of tea polyphenols in tea infusion could reduce 19.5%, and the bitterness and astringency index decreased from 3.70 to 2.98. The research provided a technological method to reduce the bitterness and astringency of purple tea leaves. By using the polyphenol-protein complex reaction model, the relative precipitation rate (R_A) was used as the index to evaluate the inhibiting ability of casein to the bitterness and astringency of polyphenols.

Study on the Analytical Method of EGCG 3″Me by HPLC in Tea

SUN Ye-liang, LV Hai-peng, LIN Zhi, LI Xing-hui

Journal of Tea Science. 2009, 29(5): 379-384. doi:10.13305/j.cnki.jts.2009.5.008

Abstract ( 471 )

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EGCG3″Me was isolated by preparative HPLC system from tea and identified by NMR and LC/MS. The influence of different extracting method on the EGCG3″Me content in tea was discussed, and a simple high performance liquid chromatography analytical method of EGCG3″Me was developed. Results showed that the analytical method showed a good linear relationship between the peak area and content of EGCG3″Me (R²=0.9998) and high reproducibility. The average recovery rate of EGCG3″Me was 95.48% and the relative standard deviation(RSD)was 3.31%. The EGCG3″Me concentration in tea infusion can reach highest when the tea sample was immersed in distilled water for 5 min at 90℃.

Study on the Characteristic of Aroma Components in Green Tea with Chestnut-like Aroma

YE Guo-zhu, JIANG Yong-wen, YIN Jun-feng, YUAN Hai-bo, ZHANG Rui-lian, WANG Zhi-lan, SHEN Dan-yu, WANG Fang, CHEN Jian-xin

Journal of Tea Science. 2009, 29(5): 385-394. doi:10.13305/j.cnki.jts.2009.5.009

Abstract ( 611 )

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E Mei MaoFeng tea with chestnut-like aroma was processed under different temperature and time to change the intensity and type of the aroma. The treated teas and E Mei MaoFeng tea were classified into two types—teas with chestnut-like aroma and others without chestnut-like aroma, according to the organoleptic evaluation. After analysis on the differences of the two types of teas using Duncan’s multiple tests, principal component analysis was applied to observe and analyze the distribution of the selected aroma components and the tea samples. The results demonstrated that the teas with chestnut-like aroma included remarkably high contents of β-ionone, nerolidol, phytol, 1,4-eicosadiene, 5,8,11,14-eicosatetraenoic acid, ethyl ester, (all-Z)-, 2,6-di-tert-butylbenzoquinone, pentadecane,2-methyl-, heptadecane, hexadecane,2,6,10-trimethyl-, but the content of dodecanol,3,7,11-trimethyl- was remarkably lower than the later. The ten aroma components have a good effect of distinguishing the teas with chestnut-like aroma from those without chestnut-like aroma. On the other hand, nonanal, and cis-jasmone are also worthy of attention.

Research on Non-Destructive Measure of Moisture Content for Packaged Tea Based on the LC Resonant Sensor

HAN An-tai, GUO Xiao-hua, CHEN Zhi-qiang, SUN Yan-wei

Journal of Tea Science. 2009, 29(5): 395-401. doi:10.13305/j.cnki.jts.2009.5.010

Abstract ( 503 )

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The design of a wireless, passive LC resonant sensor based on classic circuit theory, and its application for quick, non-destructive measured of moisture content of packaged tea were described in this paper. The proposed LC resonant sensor is an inductive-capacitive resonant circuit, which is made of a single planar capacitor and spiral inductor printed on a humidity-sensitive insulating medium substrate. To measure the moisture content of packaged tea, the LC resonant sensor is embedded inside the tea package. As the variation of the humidity inside the tea package, the humidity-sensitive insulating medium substrate absorbs the water vapor, and the planar capacitor’s capacitance and the sensor’s resonant frequency were changed, which is remotely detected through a coil connected to the instrument for measuring moisture content outside the tea package. Therefore, the moisture content of packaged tea can be indirectly determined by measuring the change in sensor’s resonant frequency. The experiment results showed that the proposed method is reasonable, and is comprised with the GB method, no significant deviation is found between the measuring results. The difficult technological problem in how to quick, accurate and non-destructive measuring moisture content of packaged tea is solved.

Experimental Analysis of Combustion Gas of Tea-seed Oil

MA Rong-chao, LI Zhang-lin, WU Wei-xiong

Journal of Tea Science. 2009, 29(5): 402-406. doi:10.13305/j.cnki.jts.2009.5.011

Abstract ( 441 )

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In the investigation on the combusting performance of tea seed oil on the S195-Diesel Engine, the exhaust experiments of degreased tea seed oil was conducted to measure its exhaust value, such as smoke, HC, NO_x, and CO. Besides, the microscopic structure of carbon particle of combustion gas was determined by the electronic scanning microscope. The results showed that it is feasible to combust degreased tea seed oil, or mixture of tea seed oil with diesel oil on the Diesel Engine with no modification of its essential structure because of its good combusting performance, less exhaust and carbon deposit.

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