15 June 2020, Volume 40 Issue 3

    

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  Research Progress on Chemical Composition and Biological Activity of Liupao Tea

  MA Wanjun, MA Shicheng, LIU Chunmei, LONG Zhirong, TANG Baojun, LIN Zhi, LYU Haipeng

  Journal of Tea Science. 2020, 40(3): 289-304. https://doi.org/10.13305/j.cnki.jts.2020.03.001

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  Liupao tea is a kind of distinctive dark tea products in Guangxi Zhuang Autonomous Region with a long history of production and marketing. It has unique flavor quality characteristics and health-care effects, which has attracted much attention in recent years. In this paper, the research progress on chemical composition and biological activity of Liupao tea over the last 20 years was reviewed, and the future research interests of Liupao tea were also discussed preliminarily.
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  Analysis of the Tea Projects Supported by National Natural Science Foundation of China from 1999 to 2019

  XIONG Xingping, ZHANG Xinzhong, YANG Zhenwei

  Journal of Tea Science. 2020, 40(3): 305-318. https://doi.org/10.13305/j.cnki.jts.2020.03.002

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  Based on the tea projects supported by National Natural Science Foundation of China (NSFC) from 1999 to 2019, a statistical analysis was carried out from the aspects of supporting year, category, department, nature of the supporting units, distribution of the supporting regions, project leaders and branches of disciplines. The current situation, characteristics and development trends of basic tea research in China were explored and the existing problems and shortcomings were discussed and analyzed. The purpose of study was to provide reference for the selection of ideas and scientific research development for tea scientists.
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  Analysis of Arbuscular Mycorrhizal Fungal Community Structure in the Rhizosphere of Different Tea Cultivars

  HE Fei, LI Donghua, BU Fan

  Journal of Tea Science. 2020, 40(3): 319-327. https://doi.org/10.13305/j.cnki.jts.2020.03.003

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  In order to enrich the arbuscular mycorrhizal (AM) fungal germplasm resources of tea plants (Camellia sinensis) in China, the community structure of AM fungi in the rhizosphere soil of different tea cultivars grown in Hanshuiyun tea garden of Ankang City, Shaanxi Province were analyzed. The results show that species richness, species and genera composition of AM fungi in the rhizosphere soil varied with tea cultivars. A total of six AM fungal species were isolated from the rhizosphere soil of Ziyang population. Likewise, five from Shancha 1, four from Longjing Changye, four from Longjing 43, and three species from Fuding Dabai. Soil collected from the rhizosphere of Longjing Changye had the highest spore density (3.57 spores per gram of dry soil), while the lowest spore density (1.10 spores per gram of dry soil) was found in the rhizosphere of Longjing 43. The highest Shannon-Wiener and Pielou evenness indices were found in the rhizosphere of Ziyang population (0.63 and 0.096), whereas the lowest values were observed in the rhizosphere of Longjing Changye (0.18 and 0.027). The maximum mycorrhizal colonization (29.5%) was found in the rhizosphere of Longjing Changye, whereas the minimum value (15.8%) was observed in the rhizosphere of Fuding Dabai. The Sorenson’s similarity coefficient of AM fungal species composition among five tested tea cultivars ranged from 0.111 to 0.750, with the highest between Longjing Changye and Longjing 43, and the lowest between Fuding Dabai and Ziyang population. The results reveal obvious differences in AM fungal community composition among the five tea cultivars. The identified AM fungal resources in rhizosphere soil are of great significance for further screening, researching AM fungi agent, and promoting the development of tea industrialization.
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  Cloning and Expression Analysis of CssHSP18.1 Gene in Camellia Sinensis

  JIANG Junmei, FANG Yuanpeng, NING Na, CHEN Meiqing, YANG Zaifu, WANG Yong, LI Xiangyang, XIE Xin

  Journal of Tea Science. 2020, 40(3): 328-340. https://doi.org/10.13305/j.cnki.jts.2020.03.004

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  The sHSPs gene family encodes a class of small molecular heat shock proteins, which are widely distributed in plants, functioned as molecular chaperones, and play an important role in plant resistance to stresses. In this study, the open reading frame (ORF) of CssHSP18.1 gene cDNA was obtained by gene cloning, which is 480 bp in length and encodes 159 amino acids. Bioinformatics analysis showed that CssHSP18.1 protein contained a typical HSP20 domain. Its molecular weight and isoelectric point are about 18.25 kDa and 5.68 respectively. Phylogenetic tree analysis showed that CssHSP18.1 has the closest relationship with quercus and apple. It was predicted that CssHSP18.1 protein was does not have signal peptide and transmembrane structure. RT-qPCR analysis showed that the expression of CssHSP18.1 under D-Mannitol treatment was lower than that in the control group. GABA could enhance the expression of CssHSP18.1 with its peak at 1 h after GABA treatment. The expression of CssHSP18.1 was upregulated upon IAA and PEG 6000 treatments, and reached the peaks at 0.5 h. Thus, GABA、IAA、PEG 6000 could induce the expression of CssHSP18.1. To obtain CssHSP18.1 soluble protein, a recombinant plasmid pET-28a-CssHSP18.1 was constructed and expressed in prokaryotic system. The expression strains, induction temperatures and induction concentrations of IPTG (isopropyl- -D-thiopyranogalactoside) were optimized. The results showed that the best expression strain of CssHSP18.1 protein was BL21 (DE3), and the best induction temperature and IPTG concentration were 30℃ and 1.2 mmol·L^-1 respectively. Finally, western blot was used to verify the expression of CssHSP18.1 protein. This study provided a basis for further study on the biological function of CssHSP18.1 gene.
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  Genetic and Phylogenetic Analysis for Resources of Camellia Sinensis from Kaihua County in Zhejiang Province

  YU Shuping, XU Liyi, WU Rongmei, WANG Liyuan, WU Liyun, WEI Kang, CHENG Hao, WANG Yongqi

  Journal of Tea Science. 2020, 40(3): 341-351. https://doi.org/10.13305/j.cnki.jts.2020.03.005

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  In this study, various tea resources from Kaihua County were collected to evaluate genetic diversity and phylogenetic relationships among individuals by SSR markers. Meanwhile, suitable core marker combinations were screened to construct fingerprint map. The results show that: (1) 14 SSR markers were polymorphic in the samples. The number of alleles per SSR locus was from 3 to 6 with the mean value of 4.14, and the average number of effective alleles was 3.08. (2) each germplasm resource could be identified by using 14 SSR markers. And based on the analysis of complex loci, the value of PE-1 and PE-3 were over 0.99 and PE-2 over 0.95, respectively, when 10 core SSR loci, as a simplified combination, were successfully screened to distinguish 36 germplasm resources. (3) 36 samples were divided into three groups based on UPGMA phylogenetic tree, and it was preliminarily speculated that five combinations might have parent-child relationship in the sample group through phylogenetic analysis. Present study indicated that tea germplasms in Kaihua County displayed highly diverse genetic backgrounds and might provide useful plant resources for breeding of new cultivars.
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  Analysis of Fat Content and Fatty Acid Composition and Absolute Content in the Tea Seeds from Southern Henan Tea Germplasms

  CHANG Yali, HUANG Xiaobing, JIANG Shuangfeng, HUANG Shuangjie, SUN Mufang, LIU Wei, GUO Guiyi

  Journal of Tea Science. 2020, 40(3): 352-362. https://doi.org/10.13305/j.cnki.jts.2020.03.006

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  The kernel percentage, fat content and fatty acid composition and absolute content of the seed kernel of 42 tea germplasm materials from Southern Henan were analyzed in this paper. The results show that the kernel percentage ranged from 33.53% to 71.60%, the fatty content was determined with Soxhlet extraction, varied from 17.77% to 38.39%, totally, 21 fatty acids were identified by GC-MS, and the range of main fatty acid contents, including palmitic acid, stearic acid, cis-oleic acid, linoleic acid and α-linolenic acid, were 2.64%-5.70%, 0.21%-1.11%,7.33%-17.29%, 0.09%-8.44% and 0.01%-0.15%, respectively. Correlation analysis shows that cis-oleic acid was significantly negatively correlated with tridecanoic acid, myristic acid and cis-eicosadienoic acid, linoleic acid was significantly negatively correlated with trans-oleic acid and cis-docosahexaenoic acid. The content ratio of saturated fatty acids, monounsaturated fatty acids and polyunsaturated fatty acids of seed kernel of Shuixian 7202 was 3.08∶12.16∶1, and others’ were around 1∶2.69∶1.37. According to higer kernel percentage and fat content of seed kernel, a total of eight tea cultivars including Zhonghuang 1, Nongkangzao, Yingshuang, Echa 11, Longjing 43, Zhongxuan 8, Laoshan 3 and Huangjinye were initially screened as candidate cultivars for constructing tea garderns harvesting leaf and seed.
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  Identification and Expression Analysis of Terpene Synthesis Related Genes during the Withering of White Tea

  CHEN Xuejin, WANG Pengjie, LIN Xinying, GU Mengya, ZHENG Yucheng, ZHENG Zhilin, YE Naixing

  Journal of Tea Science. 2020, 40(3): 363-374. https://doi.org/10.13305/j.cnki.jts.2020.03.007

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  Terpenes are the important secondary metabolites in plants and play an important role in the composition of the volatile aroma of tea plants. In this study, 141 tea plant terpenoid synthesis-related genes were identified from the tea plant genome database. Their expression specificities in different tissues were analyzed. Sixteen terpene synthetic genes which were highly expressed in the apical buds and young leaves of tea plants were screened. The results of bioinformatics methods show that the phylogenetic relationship divides the genes related to terpene synthesis of tea plant, Arabidopsis and grape into four subfamilies. The terpenoid synthesis related genes contain 5 to 14 exons and a large number of cis-related elements closely related to light response, plant growth and development, hormone and stress response according to the upstream promoter region analysis. Fluorescence quantitative detection showed that the expressions of CsMVK, CsDXS and CsGGPS were significantly up-regulated during the withering process of white tea. The expressions of CsDXR, CsMCT, CsCMK, CsMCS, CsHDS, CsGPPS and CsGGPPS showed the highest expressions at 4 h and 24 h after withering. The results of this study provided a theoretical basis for further exploring the functions of terpenoid synthesis related genes in tea.
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  Parameter Optimization and Experimental Study of Tea Twisting Machine Based on EDEM

  LI Bing, LI Weining, BAI Xuanbing, HUANG Jianhong

  Journal of Tea Science. 2020, 40(3): 375-385. https://doi.org/10.13305/j.cnki.jts.2020.03.008

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  To solve the problems caused by unstable quality in the process of tea twisting, the 6CR-40 tea twisting machine was taken as the research object in this study. The three-dimensional modeling of tea twisting machine was established based on Solidworks. The numerical simulation of the twisting process of tea twisting machine was carried out by the discrete element simulation software EDEM. The influences of various experimental factors on the performance indexes of tea twisting machine were obtained. The quadratic orthogonal rotation test was carried out and the Design-Expert was used to optimize the solution and obtain the best combination of structural parameters of twisting quality. The results showed that: when the twisting barrel rotational speed, the prismatic height, twisting plate inclination angles, forming rate of tea and breaking rate of tea were 42 r·min^-1, 10 mm, 3.8°, 88.55% and 1.83% respectively, the tea twisting machine had a good quality of twisting. The results of verification test and simulation optimization were basically consistent.
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  A Preliminary Study on the Degradation Pathway of Caffeine in Tea Microbial Solid-state Fermentation

  ZHENG Chengqin, MA Cunqiang, ZHANG Zhengyan, LI Xiaohong, WU Tingting, ZHOU Binxing

  Journal of Tea Science. 2020, 40(3): 386-396. https://doi.org/10.13305/j.cnki.jts.2020.03.009

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  In order to explore caffeine degradation products and pathways under the action of microorganisms, the dominant strains including Aspergillus sydowii NRRL250, Aspergillus pallidofulvus NRRL4789, Aspergillus sesamicola CBS137324 and Penicillium mangini CBS253.31 were screened and identified during pu-erh tea fermentation. Strains were inoculated into sun-dried green tea leaves for solid-state fermentation. High performance liquid chromatography (HPLC) was used to determine caffeine, theobromine and theophylline contents to explore the effect of microorganisms on caffeine metabolism. UHPLC-QTOF-MS was used for the metabonomic analysis of Aspergillus sydowii inoculated fermentation with sterilization treatment group (ST group) and raw material group (RM group). The results show that the dominant strains such as A. pallidofulvus NRRL4789, A. sesamicola CBS137324 and Penicillium mangini CBS253.31 had no significant effects on the metabolism of caffeine and other purine alkaloids. However, caffeine content was decreased significantly (P<0.05) with a great reduction about 83.89% during the inoculated fermentation of Aspergillus sydowii. Additionally, theophylline content was increased significantly (P<0.05) and arrived to (25.03±1.17) mg·g^-1 at the end of fermentation. While theobromine content remained stable. Therefore, Aspergillus sydowii has a profound effect on caffeine degradation metabolism. Nine metabolites related to caffeine degradation were detected by UHPLC-QTOF-MS during the inoculated fermentation, Among them, theophylline, 3-methylxanthine, 1,7-dimethylxanthine and 7-methylxanthine contents were significantly increased (P<0.05) under the action of Aspergillus sydowii which were related to N-demethylation pathway of caffeine and its related metabolites. 1,7-dimethyluric acid and 1-methyluric acid were related to the oxidation pathway of caffeine-related metabolites. It can be seen that Aspergillus sydowii is the dominant strain that can degrade caffeine and has the potential ability to convert caffeine into theophylline. Under the action of spergillus sydowii, both N-demethylation and oxidation were found in caffeine degradation metabolism, and the former was the dominant.
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  Determination of Pesticide Residue in Fresh Tea Leaves and Dry Tea by Solid Extraction and Dispersive Solid Extraction Cleanup Coupled with Tandem Mass Spectrum

  YANG Jie, ZHOU Li, YU Huan, SUN Hezhi, WANG Xinru, ZHANG Xinzhong, YANG Mei, CHEN Zongmao, LUO Fengjian

  Journal of Tea Science. 2020, 40(3): 397-406. https://doi.org/10.13305/j.cnki.jts.2020.03.010

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  A method for the determination of 9 pesticide residues in fresh and dry tea was established. Residual pesticides in samples were extracted by acetonitrile, purified by Florisil/GCB solid phase extraction column, followed by absorbent of PSA and GCB as dispersed solid phase, and determined by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and gas chromatography-mass spectrometry (GC-MS). In the spiked range of 0.005-1.000 mg·kg^-1, the average recoveries of target pesticides in fresh leaves and dried tea were 70.3%-103.9%, and the relative standard deviations (RSD) were less than 20%. In the range of 0.005-2.000 mg·kg^-1, the linear relationship of target pesticides in fresh leaves and dry tea matrix was good, with r>0.995 4. The limit of quantity (LOQ) was 0.005 mg·kg^-1. The test results of actual samples showed that the method was highly sensitive and reproducible, and can meet the requirements of multi-residue detection.
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  The Intergative Effects of Epigallocatechin-3-gallate and Vitamin C on Serum Uric Acid Levels in Hyperuricemic Mice

  XU Yan, CAI Xiaqiang, XIE Qianjin, TAI Lingling, LIU Zenghui

  Journal of Tea Science. 2020, 40(3): 407-414. https://doi.org/10.13305/j.cnki.jts.2020.03.011

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  KM male mice were used as subjects of the study. Yeast extracts (7.5 g·kg^-1) and potassium oxonate (250 mg·kg^-1) were administered to establish the hyperuricemic mice model. The study aimed to investigate the integrative effect of EGCG and vitamin C (Vc) on serum uric acid levels of hyperuricemic mice. Mice were randomly divided into 6 groups (n=6): blank group, model group, allopurinol group, EGCG group, EGCG + Vc group and Vc group. The biochemical indexes of mice were measured after 7 d of continuous administration. The results show that the serum uric acid (UA), serum urea nitrogen (BUN) and creatinine (Cr) of EGCG + Vc group were significantly lower than those of the model group (P<0.001). The combination of EGCG and Vc could remarkably inhibited the activities of adenosine deaminase (ADA) and xanthine oxidase (XOD) in the liver (P<0.05 or P<0.01) and significantly down-regulated the expression of glucose transporter 9 (GLUT9) in the kidney (P<0.001). The results of kidney slices indicated that EGCG + Vc could obviously improve the damages to the kidney in hyperuricemic mice. In addition, the integrative effect of EGCG + Vc on hyperuricemic mice was better than that of EGCG.
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  Study on Pattern Evolution and Spatial Agglomeration of Tea Production: A Case Study of Guangdong Province

  HUANG Xiujie, YAO Fei, MA Li, CHU Xialing

  Journal of Tea Science. 2020, 40(3): 415-429. https://doi.org/10.13305/j.cnki.jts.2020.03.012

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  Analyzing the evolution characteristics and agglomeration effect of tea production pattern is of great significance to the planning and layout of tea industry in Guangdong Province. In this study, the spatial gravity center model was introduced. The pattern evolution process and characteristics, and the spatial agglomeration effect of tea production in Guangdong Province were analyzed by using GIS technology and spatial autocorrelation analysis method. Results show that: (1) The planting area and yield of tea in Guangdong Province increased steadily from 1992 to 2017, and the growth rate was more obvious after 2008. (2) There were significant spatial differences in tea production in Guangdong. The northern and eastern Guangdong accounted for more than 85% of the planting area, and more than 83% of yield in Guangdong. The reduction in western Guangdong and the Pearl River Delta was obvious. (3) The center of gravity of tea production in Guangdong Province tended to move eastward and northward. The eastward shift of gravity center of tea planting area and yield reflected that the tea production has been gradually concentrated in east and north of Guangdong. (4) The spatial polarization and spillover of tea production in Guangdong Province were significant. The tea production agglomeration areas in Raoping, Chaoan, Dapu, Fengshun, Wuhua, Xingning, Yingde and Dongyuan were formed, which constituted the ‘hot spots’ of tea production in Guangdong Province, and they had stimulating effects on surrounding counties and cities. (5) Geographical environment and other natural factors were the basis for the expansion of area, the incentive and support of government policy was an important driving force for the formation of tea industry, the huge market consumption power was the direct factor of the rapid development of tea industry, the application and popularization of new cultivars and technologies were the important reasons for the expansion of tea planting area. The results indicate that the spatial agglomeration effect of tea production in Guangdong Province needs to be further strengthened. Next, it is necessary to promote the clustering development of tea production according to regional natural resources, geographical conditions and planting traditions, so as to enhance the market competitiveness of tea in Guangdong Province.