25 March 2006, Volume 26 Issue 1

    

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  Agrobacterium Rhizogenes-Mediated High Frequency Hairy Root Induction and Genetic Transformation in Tea Plant

  ZHANG Guang-hui, LIANG Yue-rong, LU Jian-liang

  Journal of Tea Science. 2006, 26(1): 1-10. https://doi.org/10.13305/j.cnki.jts.2006.01.001

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  An efficient protocol for the establishment of transgenic high hariy root tea plant (Camellia sinensis) system induction infected by Agrobacterium rhizogenes is reported using two wild type strains of A. rhizogenes, three different co-cultivation media and six different explants. The highest induction frequency was obtained from stem segments of aseptic seedlings infected by A. rhizogenes and co-cultivated on YMB solid media with 100βmmol/L AS for 2βd. The hairy roots grew rapidly on LG₀ medium without phytohormone and produced a plenty of branches and root hairs. PCR confirmed that rolA, rolB and rolC have inserted into the genomic DNA of hairy root induced by wild type A. rhizogenes. Hairy roots were also induced from explants of tea plant infected by A. rhizogenes 15834 carrying pCAMBIA2301 binary vector harboring Bt gene, transgenic root were confirmed by PCR and GUS histochemical staining that the Bt exogenous gene was inserted and expressed.
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  Quantitative Detection of β-glucosidase, β-primeverosidase Gene Expressions in Different Leaves of Tea Plantby Real-Time PCR Analysis

  ZHAO Li-ping, CHEN Liang, WANG Xin-chao, YAO Ming-zhe

  Journal of Tea Science. 2006, 26(1): 11-16. https://doi.org/10.13305/j.cnki.jts.2006.01.002

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  β-glucosidase and β-primeverosidase are considered the most important genes related to made tea aroma. An absolute quantification method for gene expression in tea plant using real-time PCR analysis was established and, the expression of β-glucosidase and β-primeverosidase genes in different leaves of Longjing 43 young shoot were determined using this method. The results indicate that the fourth leaf of the ‘five and a bud’ shoot has the highest copy number of β-glucosidase mRNA, which is 2.86E+08 copies/μl, the 4^th leaf>the 3^rd leaf>the 5^th leaf>the 2^nd leaf>one and a bud. And the highest β-primeverosidase expression is in the one and a bud, corresponding to 4.31E+06 copies/μl, one and a bud>the 2^nd leaf>the 3^rd>the 4^th leaf> the 5^th leaf. The expression intensity and pattern of these genes, which are closely related to tea aroma, are different in the young shoot. The results illuminate that the new developed method could be effectively used to detect gene expression quantitatively in tea plant.
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  Data Mining for SSRs in ESTs and Development of EST-SSR Marker in Tea Plant (Camellia sinensis)

  JIN Ji-qiang, CUI Hai-rui, CHEN Wen-yue, LU Mei-zhen, YAO Yan-ling, XIN Ya, GONG Xiao-chun

  Journal of Tea Science. 2006, 26(1): 17-23. https://doi.org/10.13305/j.cnki.jts.2006.01.003

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  ：Totally 281 SSRs distributed in 246 ESTs were mined out, accounting for 17.68% of 1589 ESTs updated in tea. The average length of tea plant EST-SSRs searched out is 33.06βbp and the average distance of distribution is 1/2.16βkb. The dinucleotide repeat is the dominant type with repeat motif AG/CT being the most common. 19 primer pairs for EST-SSRs were designed. After testing on the annealing temperature and the concentration of primers, dNTP and MgCl₂, a suitable PCR system was established. Under the condition of reaction system developed, the primers designed were screened against genomic DNA of Longjing 43 from which most EST-SSRs were derived, and 16 primer pairs showed the amplification,accounting for 82.4% of total primers. Then the primers showing amplification were subjected to PCR for DNAs from 10 tea plant cultivars and 10 primer sets showed polymorphisms, accounting for 62.5% of primers available. Results prove that it is an effective and feasible approach to develop SSR markers based on ESTs in tea.
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  Studies on the Optimal Irradiation for Tea Seedlings Under Greenhouse Manufacturing Administration

  GU Bao-jing, CHANG Jie, ZENG Jian-ming, WANG Li-yuan, YUAN Hai-bo, GE Ying, LIAO Jian-xiong, ZHOU Jian, CHENG Hao

  Journal of Tea Science. 2006, 26(1): 24-30. https://doi.org/10.13305/j.cnki.jts.2006.01.004

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  The photosynthesis, chlorophyll content and the growth traits of the clone tea seedling grown under different light conditions (8%, 15%, 35%, 42%, 50% and 75% ambient light) under the greenhouse were compared. The photosynthetic rate (P_n), maximum photosynthetic rate (P_nmax), apparent quantum yield (AQY) and new biomass reached their maximum values at 75% ambient light condition. The content of chlorophyll, death rate, specific leaf area (SLA) and leaf area ratio (LAR) decreased with the increase of light intensity, while the ratio of chlorophyll a/b, root emerging rate of the plant, traumatic tissue formation rate, root shoot ratio (R/S) and root mass ratio (RMR) increased as the light intensity increased. 50% and 75% ambient light were beneficial for the fast seedling-breeding to increase the light transmissivity of the green house and improve the light intensity for the seedling in the tents during cloudy days.
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  Resolution of DL-theanine by Aminoacylase and the Separation of Productions

  GUO Li-yun, LIU Yi, JIA Xiao-juan, LI Zhao-lan, JIAO Qing-cai

  Journal of Tea Science. 2006, 26(1): 31-36. https://doi.org/10.13305/j.cnki.jts.2006.01.005

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  An enzymatic method for the optical resolution of DL-theanine by aminoacylases from Cunnighamella echinulata 9980 was developed. The optimum temperature, pH and substrate concentration were: 50℃, pH7.0, and 0.5mol/L N-Ac-DL-theanine. The conversion rate against N-Ac-L-theanine was 92%. Productions were purified on JK008 ion exchange resin. The yield of L-theanine was 84.3%, [α] =8.1(c=2,H₂O), complied with JP2000。
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  Study on Separation and Concentration of Theanine in De-catechined Tea Extracts by Membrane System

  XIAO Li-zheng, XIAO Wen-jun, GONG Zhi-hua, WANG Wei, LIU Zhong-hua

  Journal of Tea Science. 2006, 26(1): 37-41. https://doi.org/10.13305/j.cnki.jts.2006.01.006

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  ：By using the material of de-catechined tea extracts in tea comprehensive processing and on the basis of selecting the best pH value of liquid for separation and concentration theanine by ultra-filtration,the effect on the yield rate and purity of theanine by 2500βDa、3500βDa、5000βDa ultra-filtration and four concentrating method of 300βDa nano-filtration、200βDa nano-filtration、reverse osmosis、vacuum vaporation concentration were comparatively studied in processing the de-catechined tea extracts. The results indicated that it was beneficial to separate and concentrate theanine from de-catechined tea extracts when the value of pH was 2.8~3.5.3500βDa ultrafiltration had a good effect in separating and concentrating theanine ,the major tea polyphenol ,water soluble carbohydrate were preserved and the cut rate reached 89.90%、92.20% respectively, the yield rate and purity of theanine was 54.50%,8.92% respectuvely. The lost rate of theanine in concentrating of 300βDa nano-filtration、200βDa nano-filtration、reverse osmosis、vacuum vaporation concentration was 4.51%、3.62%、0.45%、5.15% respectively. Take all thing into account, it is feasible to separate and concentrate theanine from de-catechined tea extracts in tea comprehensive processing industry by the joint application of ultra-filtration and reverse osmosis,and the yield rate and purity of theanine can reach 54.05% and 8.53% respectively.
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  Study on the Discipline System of Tea Culture

  ZHU Hong-ying

  Journal of Tea Science. 2006, 26(1): 42-48. https://doi.org/10.13305/j.cnki.jts.2006.01.007

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  From the viewpoint of professional education, the paper aims at the framework of tea culture teaching system. Firstly, the subject of tea culture has a distinct research object. Secondly, it analyzes the related research fields, which consist of the discipline of tea science, the discipline of tea art, the discipline of tea literature, the discipline of tea economy. Finally, it explores the development of tea culture on the basis of the value system. This paper tries to present a concept and model on the teaching of tea culture.
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  Study on Blood Glucose-Depressing and Blood Lipid-Depressing Function of Pu’er Tea by High Throughput Screening Method

  ZHANG Dong-ying, LIU Zhong-hua, SHI Zhao-peng, LIU Ya-lin, FU Dong-he

  Journal of Tea Science. 2006, 26(1): 49-53. https://doi.org/10.13305/j.cnki.jts.2006.01.008

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  Discovering the active substances of Pu’er Tea on blood glucose depressing and blood lipid-depressing function was studied by high throughput screening method. In this experiment, SMMC-7721 cells were used as the model, the screening concentration of samples was established at to 50βμg/ml, and the activity of Pu’er Tea on activing PPARα、PPARδ、PPARγ was screening. The result showed that Pu’er Tea ethanol extract can active PPARδand may be have the function of lowering blood glucose and blood lipid depressing function.
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  Study on Ultrasonic Wave-Assisted Extraction of Tea

  XIAO Wen-jun, TANG He-ping, GONG Zhi-hua, XIAO Li-zheng, LI Shi, LIU Zhong-hua

  Journal of Tea Science. 2006, 26(1): 54-58. https://doi.org/10.13305/j.cnki.jts.2006.01.009

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  The extraction effect of tea quality components by the ultrasonic wave-assisted extraction and the traditional water extraction was comparatively studied, and the best technical parameters for ultrasonic wave-assisted extraction was screened out. The results showed that tea extraction by ultrasonic wave-assisted extraction is better than that by traditional water extraction under the same technical parameters condition, which are ratio of tea to water, extraction temperature, time and times. The best operating parameters of tea extraction by ultrasonic wave-assisted extraction are 1:15 ratio of tea to water, 95℃water temperature, two times extraction (1:8+1:7) and 15βmin each time, and the extracted rate of amino acid, water soluble protein, polyphenol, water soluble carbohydrate, caffeine, simple catechin, estered catechin, water extracts can increase 23.13%、36.92%、14.88%、24.40%、19.24%、26.45%、10.12%、12.27% in sequence.
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  Study on Preparation and Stability of Sodium Zinc Chlorophyllin From Tea Fresh Leaves

  LAI Hai-tao, HUANG Wan-qin

  Journal of Tea Science. 2006, 26(1): 59-63. https://doi.org/10.13305/j.cnki.jts.2006.01.010

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  Sodium zinc chlorophyllin was extracted from tea leaves,and its optimal technological condition and stability were studied. The results showed that the optimal zinc reaction condition was: the weight ratio of tea powder to ZnSO₄·7H₂O is : 5:7.1, the reaction temperature is 60℃,the reaction time is 2βh,the yield of the pigment is 2.1%. In addition, sodium zinc chlorophyllin has stable character on wate- solubilization, heating, oxidizing, reducing, raying(pH:2.8~12.78),and it would not be influenced by some common food additives and metal-salts.
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  Effects of Wet-Scenting Process and RJF on the Aroma Constituent of Jasmine Scented Tea

  YE Nai-xing, YANG Guang, ZHENG Nai-hui, YANG Jiang-fan, WANG Zhen-kang, LIANG Xiao-xia

  Journal of Tea Science. 2006, 26(1): 65-71. https://doi.org/10.13305/j.cnki.jts.2006.01.011

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  The aroma constituent of high-quality jasmine-scented tea was analyzed using SPME/GC-MS method. It was found that cis-3-hexenyl acetate, linalool, benzyl acetate and methyl anthranilate were the major fragment components in jasmine scented tea. The contents of cis-3-hexenyl acetate, linalool, methyl anthranilate, cadinene, indole, benzyl alcohol, nerolidol, geranyl acetate were significantly increased in the jasmine scented tea made by wet-scenting process. The content of aromatic components in jasmine scented tea was increased when the rate of jasmine flower to tea (RJF) was ranged from 69% to 115%. When RJF is above 96%, the content of major aromatic components in the jasmine scented tea did not increase significantly. It suggested that the absorbance of tea to the fragment of jasmine flower would be attained a balance in the RJF. It suggested that the suitable RJF is 90% to 100% for producing high-quality jasmine tea made by wet-scenting process.
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  Suppression Effect of Apanteles spp Attracted Together by Infochemicals-Luring Preparation on Ectropis oblique Larvae

  HAN Bao-yu, ZHOU Peng, FU Jian-yu, CUI Lin

  Journal of Tea Science. 2006, 26(1): 72-75. https://doi.org/10.13305/j.cnki.jts.2006.01.012

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  Spectrophotometric Determination of Aluminium in Tea Leaf Samples

  LI Jing-mei, YU Hui, JU Dian-min, WANG Yi

  Journal of Tea Science. 2006, 26(1): 76-78. https://doi.org/10.13305/j.cnki.jts.2006.01.013

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