Journal of Tea Science

Research Advance on β-Glucosidase of Tea Plant

ZHOU Hanchen, LEI Pandeng, DING Yong

Journal of Tea Science. 2016, 36(2): 111-118. doi:10.13305/j.cnki.jts.2016.02.001

Abstract ( 518 )

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All material metabolisms, energy transfer, growth and development in tea plant require the participation of enzymes, which are also of great importance for tea quality formation. β-glucosidase catalyzes the hydrolysis of tea glycoside precursors. The produced aroma substances are not only involved in plant defense reaction against diseases and pests, but also important for tea quality. The β-glucosidase of Camellia sinensis is highly similar to β-glucosidase gene family members of arabidopsis, rice and maize in expression sites, subcellular localization, protein structure and phylogenetic evolution. Under biotic and abiotic stresses, the β-glucosidase of Camellia sinensis is immediately induced in the early stage. This paper mainly focused on the bio-informatic and gene expression in tea plant studies of Camellia sinensis β-glucosidase. Meanwhile, the changes of β-glucosidase in tea processing were also reviewed.

Research Advance in Synthesis and Pharmacological Effects of EGCG Derivatives

LIU Min, RAO Guowu, HUA Yunfen

Journal of Tea Science. 2016, 36(2): 119-130. doi:10.13305/j.cnki.jts.2016.02.002

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(-)-Epigallocatechin-3-gallate (EGCG), one of the important catechins in tea, has efficient bioactivity of cell experiment in vitro and animal models in vivo. However, it was not being fully utilized because of the disadvantages of poor liposolubility and stability, as well as low bioavailability, which was resulted from its ‘polyhydroxy’ chemical structure. Excitingly, the molecular modification would be used to improve the physicochemical character of EGCG. This review will summarize the methods on molecular modification of EGCG and investigation of biological activities, which hopes to provide worthful references to the further study of EGCG.

Genetic Diversity Analysis of Tea Germplasm in Shaanxi Province Based on SCoT Marker

CHEN Xi, ZHANG Yu, LI Jiao, XI Yanjun, ZHANG Yanqing

Journal of Tea Science. 2016, 36(2): 131-138. doi:10.13305/j.cnki.jts.2016.02.003

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Genetic diversity of 50 tea germplasm materials in Shaanxi was investigated. Taking 6 representative Shaanxi tea germplasm materials as templates, 38 of 121 SCoT marker primers were screened out, which showed specific bands and highly polymorphism. The results showed that SCoT markers could reveal high genetic diversity among tea germplasm, and each primer can detect out 4 to 17 alleles. On average, 11 allelic variations could be detected by each SCoT marker. 414 DNA fragments were amplified via 38 pairs of SCoT primer, of which 400 showed polymorphism and the ratio (PPB) was 96.62%. In addition, genetic polymorphism information content of each SCoT locus was between 0.56 and 0.99, the average of which was 0.90 for the tested materials, and the genetic similarity coefficient was between 0.59 and 0.71. The research showed that SCoT marker could be used in the genetic diversity research of tea germplasm. Basing on SCoT marker, genetic diversity level of these Shaanxi tea germplasms were relatively high in the genetic diversity.

Design and Process Optimization of a Continuous Production Line on Needle-type Premium Green Tea

CHEN Gensheng, YUAN Haibo, XU Yongquan, JIANG Yongwen, DENG Yuliang, YU Liaoyuan, YU Shuping, YIN Junfeng

Journal of Tea Science. 2016, 36(2): 139-148. doi:10.13305/j.cnki.jts.2016.02.004

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According to the quality and processing characteristics of needle-type premium green tea, a strongly practicable, energy-efficient and highly qualified needle-type premium green tea continuous production line which consisted of the fresh tea leaves artificial spreading machine, electromagnetic heat roller fixation machine, multiphase continuous fixation-shaping machine, rubbing machine, far-infrared raising fragrant machine and other energy efficient facilities was organized. The processing procedures of the continuous production line including spreading, fixation, fixation-shaping and baking were investigated and a complete set of processing technology about the line on needle-typepremium green tea was proposed. The results showed that the best process parameters were: temperature at 18-25℃ and humidity at 60%-90% for fresh tea leaves spreading, the leaf weight of 125 kg·h^-1, baking temperature at 270℃ and time of 65 second for the fixing, 240℃(38 Hz）, 210℃(40 Hz) and 180℃(45 Hz) for the multiphase continuous fixation-shaping, and drying temperature at 40-55℃, baking temperature at 100℃, speed of 30 kg·h^-1 for the rotary continuous drying.

Effect of Light Intensity on Dynamic Changes of Main Biochemical Components of Tea Fermented Leaves

YU Luting, YUAN Haibo, WANG Weiwei, CHEN Gensheng, HUA Jinjie, JIANG Yongwen

Journal of Tea Science. 2016, 36(2): 149-159. doi:10.13305/j.cnki.jts.2016.02.005

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By using two leaves and a bud shoot of Fuding Dabai as fresh leaves material, the dynamic effects of six different fermenting light intensities (0 lx, 3 000 lx, 6 000 lx, 9 000 lx, 12 000 lx and 15 000 lx) on major biochemical components of tea fermented leaves were studied. Tea samples were collected every 30 min and the contents of tea polyphenols, flavonoids, total amino acids, soluble protein, soluble sugar, catechin compositions, theaflavins and amino acids were detected. The results showed that with the increasing of fermentation time, the contents of tea polyphenols and catechins compositions decreased gradually. The contents of flavonoids, total amino acids and soluble sugar showed opposite trends. Both the soluble protein and theaflavins components increased at first and then dropped, with its peak at around 120 min. Comparing different light intensity processings, 15 000 lx strong light treatment significantly decreased the contents of tea polyphenols, catechins compositions, the accumulation of flavonoids and the amino acid compositions. The highest contents of soluble protein and soluble sugar were identified in samples under 9 000 lx light treatment. While, the highest contents of theaflavins components and theanine were found in samples under 3 000 lx light treatment. The sensory evaluation also showed that the liquor color and taste of samples under 3 000 lx light treatment got the highest score, indicating a good quality.

Biological Characterization of Fungi Involved in Fu Brick Tea Fermentation

ZHAO Renliang, TAN Jihui, LU Qinhua, WU Dan, ZHU Qi

Journal of Tea Science. 2016, 36(2): 160-168. doi:10.13305/j.cnki.jts.2016.02.006

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It was assessed that the growth characteristics and succession rules of fungi involved in the fermentation of Fu brick tea during the winter and summer seasons in Hunan Province, China. 27 fungal strains were isolated and identified, and it was investigated that the growth and sporulation on different media, different carbon and nitrogen sources, and under different temperatures. The results showed that all fungi were able to grow on culture medium containing tea juice. Mycelium grew faster and were more prone to spore production in culture media with monosaccharides or disaccharides as the carbon source and organic nitrogen as the nitrogen source. The optimum growth temperature ranged at 25-30℃ for most fungi. The temperature higher than 30℃ or lower than 25℃ inhibited the growth of fungi. However, there were some differences between strains, it meant the different stain and succession rules of fungal strains during Fu brick tea fermentation in different seasons were closely related to the biological characteristics of fungi.

The Investigation of the Protection Effects and Mechanism of EGCG on Kidney Ischemia Reperfusion Injury

LIU Hongguo

Journal of Tea Science. 2016, 36(2): 169-174. doi:10.13305/j.cnki.jts.2016.02.007

Abstract ( 415 )

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The study aims to investigate potential protection and mechanism of epigallocatechin gallate（EGCG） on kidney ischemia reperfusion injury. The results showed that EGCG treatment improved renal dysfuction, pathlogical change of kidney tissues, and reduced the expression of Wnt3a, β-catenin, MDA, IL-6, IFN-γ and TNF-α, but increased the levels of CAT, GP_X and SOD, suggesting that EGCG can protect the kidney from ischemia-reperfusion injury in rats which is associated with suppressing the activation of Wnt/β-catenin/p53 signaling pathway and inhibiting inflammation and oxidative stress.

Analysis of Aroma Components in Big-leaf Tea Plant (Camellia sinensis var. assamica) Flower Using Headspace Solid-phase Microextraction Coupled with GC/MS

WANG Chen, LÜ Shidong, LIAN Ming, MENG Qingxiong

Journal of Tea Science. 2016, 36(2): 175-183. doi:10.13305/j.cnki.jts.2016.02.008

Abstract ( 654 )

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The volatile components of tea flower (Camellia sinensis var. assamica), which was collected from tea garden in Pu-erh district, were extracted by fully automated headspace solid-phase microextraction (HS-SPME), identified by gas chromatography-mass spectrometry (GC/MS), and compared with the volatile components of Pu-erh green tea from same tea garden. The results showed that 9 and 67 kinds of volatile components were identified in tea flower and Pu-erh raw tea, respectively. Among them, hydrocarbons and alcohols were two major volatile components. Comparison showed that all volatile components of the tea flower were also found in Pu-erh green tea. Our results suggests that the tea flower may have similar aroma to tea leaves, and can also be mixed with tea leaves to improve the aromatic flavor.

Nondestructive Measurement of Moisture of Black Tea in Drying Process Based on Near Infrared Spectroscopy

CHEN Lin, DONG Chunwang, GAO Mingzhu, YE Yang

Journal of Tea Science. 2016, 36(2): 184-190. doi:10.13305/j.cnki.jts.2016.02.009

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Moisture is an important index of tea drying effect and quality. To understand rapid detection of moisture in black tea, a nondestructive testing method was proposed based on near infrared spectroscopy (NIR). The diffuse reflectance spectra of 226 tea samples were scanned in the range of 1 000-1 799 nm. These samples were from 6 drying processes. Moisture contents of samples were immediately measured after spectral scanning. The original spectrum data were proposed by the Standard Normal Variate Transformation (SNVT). Two regression algorithms including Partial Least Square (PLS) and Synergy Interval Partial Least Square (siPLS) were used to develop models for determination of moisture contents respectively. The result showed that both models had high accuracy, but the performance of model with siPLS was better. It contained 13 spectral intervals combined with 4 subinterval and 6 principal component factors. The root mean square for prediction (RMSEP) and the correlation coefficient (Rp) reached 0.0395 and 0.9593, respectively. It showed that it is feasible to measure moisture content of black tea during drying process.

Contents of Glomalin-related Soil Protein and Its Correlations with Soil Factors in the Rhizosphere of Tea Plant [Camellia Sinensis(L.) O. Kuntze]

GAO Xiubing, XING Dan, CHEN Yao, ZHOU Fuyu, ZHAO Huafu, CHEN Juan, GUO Can, ZHOU Yufeng

Journal of Tea Science. 2016, 36(2): 191-200. doi:10.13305/j.cnki.jts.2016.02.010

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Glomalin-related soil protein (GRSP) is a kind of glycoprotein containing metal ions that are secreted by arbuscular mycorrhizal fungi (AMF). It plays an important role in maintaining physiological functions of AMF and stabilization of soil organic carbon and soil aggregate. In this paper, the contents of GRSP and its correlations with soil factors were analyzed in the rhizosphere of tea plant [Camellia Sinensis(L.) O. Kuntze]. Total glomalin-related soil protein (T-GRSP) and easily extractable glomalin-related soil protein (EE-GRSP) as well as soil factors were investigated by Bradford and conventional soil factors analysis method in four key tea planting areas (Meitan county Shiqian county Guiding county and Douyun city) of Guizhou province southwest China. The results showed that GRSP levels were dependent on both rhizospheres of different tea cultivars and planting areas. The range of T-GRSP and EE-GRSP were 5.71-22.84 mg·g^-1 and 2.35-7.91 mg·g^-1; with average of 12.96 mg·g^-1 and 4.88 mg·g^-1 respectively. Correlation analysis showed that T-GRSP was significant positive correlation with hydrolysable nitrogen (N_h); organic matter (OM) and available potassium (P_a). EE-GRSP was positive correlated with N_h P_a and OM but negative correlated with soil pH. The result of multiple linear regression equation showed that different soil factors showed different impact on the content of T-GRSP and EE-GRSP. For T-GRSP: N_h > P_a > OM. For EE-GRSP: OM > P_a. These results showed the GRSP was remarkably abundant in the rhizosphere of tea plant and closely related with soil factors; so the content of GRSP may be an appropriate index for evaluating soil quality in tea garden.

Inhibition of Biofilm Development and Spoilage Potential in Shewanella baltica by Epigallocatechin Gallate

YE Xiaofeng, ZHANG Fang, ZHU Junli, ZHANG Lei, XIE Dumei

Journal of Tea Science. 2016, 36(2): 201-209. doi:10.13305/j.cnki.jts.2016.02.012

Abstract ( 451 )

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The anti-biofilm activity of catechin monomers from tea polyphenols against S. baltica, as the specific spoilage organism in marine product, was investigated. Effects of catechin monomer with the highest inhibitory activity on biofilm formation and spoilage potential of S. baltica were further assessed. The results showed that all five monomer of catechins significantly repressed biofilm formation of S. baltica, in which epigallocatechin gallate (EGCG) exhibited the highest activity. The minimum inhibitory concentration (MIC) of EGCG against S. baltica was 40 µg·mL^-1, and EGCG at the sub-inhibitory concentration of 10-30 µg·mL^-1 had no effect on its growth. EGCG at 30 µg·mL^-1 inhibited significantly the activity of quorum sensing (QS) autoinducers, AI-2, cyclo-(L-Pro-L-Leu) and cyclo-(L-Pro-L-Phe) (P<0.05), with reduction by 63.95%, 27.43% and 21.94% respectively. The biofilm development, swimming motility and protease activity also decreased considerably after incubation with sub-MIC of EGCG, and exhibited a dose dependent effect, which presented the reduction by 55.18%, 66.71% and 46.67% in the presence of 30 μg·mL^-1 EGCG. Hence, our results indicated that the sub-MIC of EGCG could interfere with S. baltica QS system, and inhibit the biofilm formation and spoilage potential of S. baltica. The present study suggested the potential of EGCG as QS inhibitor in controlling food spoilage bacterial.

An Efficient Identification of Tea Cultivars in Fujian with A Strategy of Cultivar Identification Diagram(CID) Based on Fluorescent Labeled SSR Markers

WANG Rangjian, YANG Jun, KONG Xiangrui, GAO Xiangfeng

Journal of Tea Science. 2016, 36(2): 210-218. doi:10.13305/j.cnki.jts.2016.02.013

Abstract ( 499 )

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The authenticity identification of tea cultivars is an important part of the research on tea germplasm resources, and the precondition of intellectual property protection of tea cultivars, 45 SSR markers were selected to synthesize fluorescent labeled SSR primer pairs based on the principle of uniform distribution in the genome of the tea plant. Using these markers, 54 clonal tea cultivars in FUJIAN province were amplified by PCR and 9 SSR markers were selected as the core primer pairs after genotyping. The polymorphism information content (PIC) of the core primer pairs was more than 0.5, and the number of all alleles was varied from 3 to 5, the number of genotypes was varied from 5 to 10. These core primer pairs were suitable for identifying the authenticity of tea cultivars for 54 Fujian clonal tea cultivars due to high polymorphism and convenient to genotype. The cultivar identification diagram (CID) was then established using 6 of these core primer pairs. The CID provided primer pairs and its corresponding genotype, by which any cultivar in the CID could be identified. The capacity of the CID can be expanded so as to facilitate the authenticity identification of more clonal tea cultivars. The method was considered to be easy, fast, accurate, stable and practical for the authenticity identification of tea cultivars. The CID is important for the protection of intellectual property of tea cultivars, and will be helpful for the healthy and sustainable development of tea breeding.

Cloning and Bioinformatical Analysis of Anthocyanin Synthase Gene and Its Promoter in Camellia sinensis

JIN Qifang, CHEN Zhidan, SUN Weijiang, LIN Fuming, XUE Zhihui, HUANG Yan, TANG Xiuhua

Journal of Tea Science. 2016, 36(2): 219-228. doi:10.13305/j.cnki.jts.2016.02.014

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The full length cDNA sequence of anthocyanin synthase gene（CsANS）was cloned from ‘Wuyi qizhong C18’ (Camellia sinensis) using rapid amplification of cDNA ends（RACE）technology. The promoter of CsANS was isolated using Genome Walking technology. The gene expression of ANS under different shading treatments was analyzed by the real-time PCR. The results showed that the full length cDNA of CsANS was 1 000 bp, and it contained a 957 bp open reading frame (ORF), which encoding a protein of 320 amino acid, including two conservative functional domains, DIOX-N and 20G-Fell-Oxy. A sequence containing 1 010 bp was isolated and found to be the promoter of CsANS, which contained many cis-acting elements related to anthocyanin biosynthetic pathway, such as the core promoter element TATA-box and light responsive element (including ACE, GT1-motif and Sp1), circadian (cis-acting regulatory element involved in circadian control). The real-time PCR analysis revealed that the gene was highly expressed under the sunshine (CK), while lowly expressed under the 75% shading. This study reveals that the expression of CsANS was regulated by light intensity.

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