Journal of Tea Science

Research Progress in Genetic Map of Tea Plant (Camellia sinensis)

MA Jian-qiang, YAO Ming-zhe, CHEN Liang

Journal of Tea Science. 2010, 30(5): 329-335. doi:10.13305/j.cnki.jts.2010.05.001

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Conventional breeding of tea plant is laborious and time-consuming, due to long generation, self-incompatibility and high heterozygosity. A saturated genetic map is a vital tool in the analysis of genome structure, genetic evolution, QTLs (Quantitative trait loci), gene cloning and MAS (Marker assisted selection), which may provide great potential for accelerating cultivar improvement programme in tea plant. This paper introduced the construction theory and procedure of genetic mapping, as well as research progress and problems encountered by previous workers in genetic map construction of tea plant. Finally, we proposed improved strategies for the construction of genetic map of tea plant.

Extraction and Analysis on the Aromatic Components of Pu-erh Ripe Tea and Raw Tea

HONG Tao, HUANG Zun-xi, LI Jun-jun, TANG Xiang-hua, MU Yue-lin, XU Bo

Journal of Tea Science. 2010, 30(5): 336-342. doi:10.13305/j.cnki.jts.2010.05.002

Abstract ( 546 )

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The aromatic components from Pu-erh ripe and raw tea which storaged for different years extracted by methylene chloride and ethyl ether through vacuum distillation extraction, and then investigated by GC-MS. It showed that the aromatic compounds extracted by methylene chloride were more than ethyl ether no matter they were ripe tea or raw tea. Those compounds including hydrocarbons, heterocyclic oxygen compounds, nitrogenous compounds, ketones were extracted more by methylene chloride than those extracted by ethyl ether. However, more esters and phenolic compounds were extracted by ethyl ether. The contents of benzothiazole and caffeine increased during the process of ripe tea’s storage, but the contents of phthalic acid, isobutyl octyl ester and 1,2,4-trimethoxybenzene were decreased. The contents of 5-Amino-1-ethylpyrazole, N-phenyl-1-naphthalenamine, 1,2,3-trimethoxybenzene and phthalic acid-iso-butyl-octyl ester were increased during the storaging process of raw tea, but the contents of trans-squalene, linalool, β-iso-methyl ionone and caffeine were decreased. At the same time, there were many aromatic compounds of heterocyclic oxygen compounds in ripe tea. The main components of these 10 heterocyclic oxygen compounds included 1,2,3-trimethoxybenzene. The contents of alclhols aromatic components in raw tea, such as 1-octen-3-yl, linalool, cyclohexanol, 2,6-dimethylcyclohexanol, phenethyl alcohol and cedrol.

Effect of Selenium-Zinc Interaction on the Chemical Form of Selenium of Tea Leaves in Spring Season

LIU Yang, WANG Chang-quan, LI Bing, LI Huan-xiu, DU Qian, ZHANG Wei

Journal of Tea Science. 2010, 30(5): 343-349. doi:10.13305/j.cnki.jts.2010.05.003

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Foliar spraying in different levels of Na₂SeO₃ and (CH₃COO)₂Zn·2H₂O was carried out to determine the effects of Se, Zn and their interaction on the chemical form of Se in tea leaves in spring season. The results showed that: (1) The content of Se in alcohol soluble, water soluble, salt soluble was increased when spraying Na₂SeO₃, particularly the content of Se of three kinds of chemical forms was increased significantly when the spraying concentration of Se was 200~400 μg/mL. Among there the proportion of the total alcohol soluble, Se was the highest and water soluble was higher than salt soluble, the proportion of the total of alcohol soluble and water soluble Se increased in varying degrees comparing with the control. (2)The cumulative effect of the content of Se of three kinds of chemical forms was obvious when the spraying concentration of Zn was between 0.20%~0.40%. The Se of three kinds of chemical forms assimilation was restrained when the zinc concentration increased to 1.2%. (3)The overall three kinds of forms of selenium levels were lower and there was no obvious difference of the proportion of the total of Se between alcohol soluble and water soluble when spraying Se at 50μg/mL and the different concentration of Zn, but both of them were higher than salt soluble Se. The distribution of chemical forms of selenium was similar to single Se treatment when the spraying with the other concentrations of Se and Zn. The content of Se of three kinds of chemical forms was increased obviously when the spraying of Se in 200~400 μg/mL and Zn in 0.4%~0.8%, it also raised the proportion of the total of alcohol soluble and water soluble Se comparing with the control, but the effect achieved the best when spraying Se in 200 μg/mL and Zn in 0.80%.

Isolation and Identification of “Jinhua” Fungi from the Loose Tea with“Fungus Growing”

HUANG Hao, LIU Zhong-hua, HUANG Jian-an, LI Shi, LI Juan, WU Yang, DENG Ting-ting, GONG-Xue

Journal of Tea Science. 2010, 30(5): 350-354. doi:10.13305/j.cnki.jts.2010.05.004

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A fungi called “Jinhua” that separated from Jing Xiangyi brick tea was produced by Hunan Yiyang Tea Factory in 2008. Inoculating the fungi to the raw material of Yiping Fuzhuan tea of 2008 with “Fungus growing” by controlling the conditions of environment. A strain of “Jinhua” fungi was isolated from the loose tea with fungus growing as a tested fungi through subculture purification. The epigamous and phorozoon of the tested fungi was observed with microscope, and the ascospores, and conidia of the tested fungi was observed SEM, and the tested fungi was identified in the molecular level by DNA sequencing. According to the characteristic of the morphology, microscopic, and SEM observed as well as the DNA sequencing of the tested strains, it was identified as Eurotium cristatum.

Size Distribution of Paticles and Concentration of Heavy Metals in Foliar Dust of Tea Production Areas

YANG Dong-wei, ZHANG Ming-kui

Journal of Tea Science. 2010, 30(5): 355-361. doi:10.13305/j.cnki.jts.2010.05.005

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To understand the concentrations of heavy metals in foliar dust in tea production areas, tea foliar dust samples were collected from five different tea production areas, including nearby urban, nearby mining, nearby highway, nearby village and a clean area without any pollution source. Size distribution of the dust samples was determined with a laser particle sizer, and total concentrations of heavy metals in the samples were determined with ICP-AES. The possible sources of the heavy metals in the dust were also discussed. It was found that the particulate size was distributed symmetrically with a range less than 1 mm. More than 76.52% of the dust was below 100 µm. Mean size of the dust increased in the sequence of a clean area < nearby village < nearby urban < nearby highway < nearby mining. Cd, Pb, Ni, Cu, Zn, and Hg were significantly enriched in the tea foliar dust, and concentrations of the metals ranged from 1.76 to 11.34 mg/kg for Cd, 87 to 648 mg/kg for Pb, 36 to154 mg/kg for Ni, 133 to 476 mg/kg for Cu, 165 to 1121 mg/kg for Zn, and 0.36 to 4.23 mg/kg for Hg, respectively. The concentrations varied with sampling areas, and were highest in the nearby mining, and lowest in the clean area. Both correlation analysis and principal component analysis suggested that Cd, Pb, Cu, Zn, and Hg in the dust were originated from industrial pollution, vehicle emission, and other anthropogenic sources, and Cr, Co, Ni, Al, Fe, and Mn could be attributed to natural sources.

Cloning and Expression Analysis on Citrate Synthase Gene of Tea Root under Low Phosphorous Stress

LIN Zheng-he, CHEN Chang-song, WU Ling-sheng

Journal of Tea Science. 2010, 30(5): 362-366. doi:10.13305/j.cnki.jts.2010.05.006

Abstract ( 471 )

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Citrate synthase(CS) is the key enzyme of producing citrate acid. CS gene fragments were cloned from tea roots. The GenBank accession is FJ814766 in the NCBI. Then the changes in gene expression in response to P supply were investigated by using semi-quantitative RT-PCR. The result showed that P deficiency increased the expressions of CS gene. This may be a tolerant mechanism of tea seedlings to P deficiency.

Construction of DNA Fingerprints for Tea Cultivars Originated from Hunan Province

YANG Yang, LIU Zhen, ZHAO Yang, LIANG Guo-qiang

Journal of Tea Science. 2010, 30(5): 367-373. doi:10.13305/j.cnki.jts.2010.05.007

Abstract ( 484 )

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Molecular fingerprints is very important to identify new varieties, cultivars authenticity and seedings purity. Seventeen pairs of SSR primers were used to construct the molecular fingerprints of tea varieties originated from Hunan province. Totally 41 alleles were amplified using the 17 SSR primers, the number of alleles per primer ranged from 2 to 3, with an average of 2.4. The SSR alleles were determined for bands size(bp) and scored as present(1) or absent (0). According to the genotypes code rule, the 1 and 0 results which one pair of SSR primers amplified in a variety could be converted to genotypes code of 1, 2……N following their fragment size. The results proved it’s a very useful tool for tea resource appraising, all accessions could be discriminated from a set of molecular fingerprints. Variety speciality index varied from 65.4 to 113.7, averagely 80.1, which indicated that there is great difference in speciality among all accessions.

Combined Effects of Caffeine and Catechins on the Body Weight and Lipid Metabolism in Mice

YANG Li-cong, ZHENG Guo-dong, WANG Chun-rong, LI Dong-ming

Journal of Tea Science. 2010, 30(5): 374-378. doi:10.13305/j.cnki.jts.2010.05.008

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To investigate the combined effects of caffeine and catechins on the body weight and lipid metabolism, mice was with on diets containing different concentration of caffeine and catechins for 12 weeks. Organs and intraperitioneal adipose tissues (IPAT) were weighted at the end of this period. Biochemical parameters in the serum and lipid levels in the liver were measured. The body weight gain was significantly reduced by group Ⅳ(0.06% caffeine+0.6% catechins). Weight of IPAT was remarkable reduced in all experiment groups. Serum concentration of NEFA was significantly reduced by group Ⅰ(0.03% caffeine + 0.3% catechins) and group Ⅱ(0.03% caffeine+0.6% catechins). Group Ⅲ(0.06% caffeine+0.3% catechins) and groupⅣ(0.06% caffeine + 0.6% catechins) decreased TC, TG, leptin level in serum. Liver TC and TG levels were reduced by combinations of caffeine and catechins. These results showed the caffeine and catechins combination might reduce lipid levels in the serum and the liver. It causes the suppressive effect on fat accumulation and body weight.

Comparation on the Inhibitory Effect of Tea Polyphenols and Rape Pollen Extract on Prostate Cancer Cell

ZHANG Xing-hai, GONG Shu, WANG Yue-fei, ZHOU Xiao-hong

Journal of Tea Science. 2010, 30(5): 379-383. doi:10.13305/j.cnki.jts.2010.05.009

Abstract ( 530 )

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To study the effect of tea polyphenols (TP) or rape pollen extract (RPE) on the inhibition of PC-3 cell in vitro. MTT assay was used to evaluate the inhibition activity (IC₅₀) on PC-3 proliferation, and fluorescence microscope was used to observe the apotosis cells. Results showed that water-soluble extract of pollen (A), ethanol-soluble pollen extract (B) and TP were proved to be effectively inhibited on the prostate cancer cell PC-3. The IC₅₀ of A, A+TP, B+TP and TP were 1470, 752, 82 μg/mL and 45 μg/mL, respectively. B+TP showed effect-dose and effect-time relationship. B+TP was also found to induce cell apotosis as shown by changes in nuclear morphology and DNA fragmentation.

Changes in Quality of Chimonanthus Tea during Processing

ZHOU Ji-rong, ZHENG Kai-ying, DI Ying-jie, CHUN Xiao-ya, NI De-jiang

Journal of Tea Science. 2010, 30(5): 393-398. doi:10.13305/j.cnki.jts.2010.05.010

Abstract ( 439 )

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Changes in organoleptic quality, color, aroma compounds and main chemistry components of chimonanthus tea during processing (tea dhool→firstly scented→secondly scented→thirdly scented→raising fragrance) were investigated using organoleptic appreciation, chromatic evaluation, SED in combination with GC/MS and spectrophotometry. Results showed that, the total score increased during processing, but it was slightly less in the thirdly scented tea than the secondly scented. The Ha*b* value of thirdly scented tea significantly decreased after thirdly scenting (P<0.05). The main aroma compounds of chimonanthus tea were benzyl acetate, 2-aminobenzoate-3,7-dimethyl-1,6- Octadien-3- ol, methyl salicylate, benzyl alcohol, linalool, alloocimene, naphthalene, α-ionone. The relative contents of esters and alcohols in the secondly scented tea were significantly the highest among samples (P<0.01), and that of ketones did not significantly change during processing, and that of terpenoids in raising fragrance tea were significantly more than that in other samples (P<0.05). The contents of polyphenols and chlorophyll in thirdly scented tea significantly decreased compared to that of dhool (P<0.05), but that of soluble sugar significantly increased after secondly scenting (P<0.05), and that of amino acids did not significantly change. These results also showed that two-time continuously scenting and one-time raising fragrance procedures could be used for chimonanthus tea production.

The Nonlinear Regression of Bitterness and Astringency of Main Compounds in Green Tea and the Application in Organoleptic Tests

XU Wen-ping, LI Da-xiang, ZHANG Zheng-zhu, TANG Qian, WAN Xiao-chun

Journal of Tea Science. 2010, 30(5): 399-406. doi:10.13305/j.cnki.jts.2010.05.011

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The methods of quantitative response scales and sequential analysis for organoleptic tests were employed to quantitatively evaluate the bitterness and astringency of tea infusions. Flavor characteristic curves (the relationship between taste intensity and concentration) of EGCG and caffeine were established. The mathematical functions of bitterness and astringency were built based on three main compounds namely EGCG (epigallocatechin-3-O- gallate), caffeine and rutin, and the interactions of the three compounds on bitterness and astringency were examined. The results showed that rutin could increase the bitterness of caffeine, but it had no effect on flavor of EGCG, while caffeine could enhance astringency of EGCG. The functions were used to discover the mysteries of heavy bitterness and astringency of green tea produced in Sichuan province. The results from the quantification of main tastants and flavor of twenty tea samples showed that the functions could give the explanation of 78 to 137 percents to the bitterness and 68 to 144 percents to the astringency. The heavy bitterness and astringency of Sichuan green tea were mainly caused by high contents of EGCG and caffeine.

Effect of Spreading on Treatments of different Fresh Tea Leaves the Aroma Quality of Green Tea Produced in Summer and Autumn

AO Cun, TANG De-song, GONG Shu-ying, LIU Lei, GU Zhi-lei

Journal of Tea Science. 2010, 30(5): 400-408. doi:10.13305/j.cnki.jts.2010.05.012

Abstract ( 446 )

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Green tea produced in summer and autumn using conventional methods is of low aroma quality because of the properties of fresh tea leaves. In this work, the method of spreading fresh leaves was employed in the processing of green tea in summer and autumn. The aroma quality of obtained green tea was sensorially evaluated and the aroma compounds were analyzed using GC/MS. The green tea produced using the treated fresh leaves obtained good aroma quality: 1) the bud tea had flowery smell while the control did not. 2) The components with flowery smell in tea samples of one bud and two or three leaves was much higher than the control. 3) After treatment the grassy smell of single leaves and banjhi shoots reduced and flowery smell appeared. The results indicated the proper methods of spreading could improve the aroma quality of green tea produced in summer and autumn.

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