Journal of Tea Science

Camellia Ptilophylla and Specific Chemical Components, Theirs Health Beneficial Effects

WU Wenliang, TONG Tong, HU Yao, ZHOU Hao, YIN Xia, ZHANG Shuguang

Journal of Tea Science. 2021, 41(5): 593-607. doi:10.13305/j.cnki.jts.20210917.002

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Cocoa tea (Camellia ptilophylla Chang) is a specific tea resource in China, and theobromine (TB) and gallocatechin gallate (GCG), as the dominant chemical components of cocoa tea, have various health effects. This paper summarized intervention effects and mechanisms of cocoa tea, TB and GCG on cardiovascular diseases, cancers, obesity, diabetes, neurodegenerative diseases, dental diseases, respiratory diseases and kidney diseases, etc. This review would provide a theoretical reference for the development of functional cocoa tea products.

Analysis on the Achievements of Tea Science and Technology Awards at Provincial and Ministerial Level and above in China from 2008 to 2019

XIONG Xingping, ZHANG Xinzhong, LI Hongbin, SUN Liang, YAO Mingzhe

Journal of Tea Science. 2021, 41(5): 608-618. doi:10.13305/j.cnki.jts.20210917.001

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Based on the national, provincial and ministerial science and technology awards in the field of tea science in China from 2008 to 2019, statistical analyses on the award, grade, year of award, the first completed person, the first completed unit, the main research fields, and the cooperation between industry, university and research unit were performed. The current status, characteristics and support trends of tea science and technology awards in China were clarified, aiming to provide ideas for Chinese tea science and technology personnel to cultivate achievement, determine scientific research topics and apply for awards.

Analysis of Genetic Diversity and Population Structure of 32 Tea Landraces in China

LI Changle, GE Yue, YAN Meilin, LI Hui, LIN Qingqing, WANG Pu, ZHAO Hua, WANG Mingle, WANG Yu, GUO Fei, NI Dejiang

Journal of Tea Science. 2021, 41(5): 619-630. doi:10.13305/j.cnki.jts.20210804.002

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Tea plant populations are natural populations of tea plants. They are cultivated in specific production areas and contain individuals with different economic and biological traits. There are great differences among individuals and there are many types. They are important materials for studying the evolutionary relationship and breeding of tea plants. In this study, 30 pairs of SSR primers distributed on 15 linkage groups were selected to analyze the genetic diversity and population structure of 32 tea plant populations from 12 provinces, in order to provide a reference for the selection of tea breeding parents and the inference of evolution route. In this study, a total of 149 alleles were obtained with an average of 5.96 for each SSR marker. The average polymorphism information content of primers was 0.660. Shannon's diversity index of 32 tea populations ranged from 0.691 to 1.089, with an average of 0.954. The observed heterozygosity ranged from 0.253 to 0.633, with an average of 0.510. The expected heterozygosity ranged from 0.430 to 0.653, with an average of 0.590. The average genetic differentiation coefficient (Fst) of tea plant population was 0.205, indicating a high level of genetic differentiation. The results of clustering based on Nei's genetic distance and population structure analysis were consistent. The germplasm to be tested was divided into 4 major types, with obvious regional distribution.

Cloning and Expression Analysis of CsWRKY17 Transcription Factor in Tea Plants

LIU Miaomiao, ZANG Liansheng, SUN Xiaoling, ZHOU Zhongshi, YE Meng

Journal of Tea Science. 2021, 41(5): 631-642. doi:10.13305/j.cnki.jts.2021.05.001

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WRKY transcription factors, a super family of plant transcription factors, play an essential role in the regulation of plant defense responses to herbivores. While the roles of herbivore-related WRKY transcription factors are well established in grass plants, their roles in woody plants are still largely unknown. Here, we cloned a WRKY transcription factor, named CsWRKY17. CsWRKY17 has a full length of 1 141 bp, contains a 987 bp open reading frame, and encodes 328 amino acids. Based on the conserved domain analysis, CsWRKY17 belongs to the WRKY Ⅱ subfamily, containing one conserved WRKY domain and a typical C₂H₂-type zinc finger motif. Homology alignment and phylogenetic tree analysis show that CsWRKY17 has the closest relationship with AtWKRY11 and AtWRKY17 in Arabidopsis thaliana. Moreover, CsWRKY17 exhibited a tissue specific expression, and was also induced by mechanical wounding, tea geometrid (Ectropis oblique) attack, simulated herbivory, and exogenous phytohormone treatments like JA. Transient expression experiments indicate that it might play a role in the nucleus. Taken together, we proposed that CsWRKY17 is a potential regulator of herbivore-induced defense responses against herbivores in tea plants through JA, ABA, GA and BR signaling. Our study paved the way for molecular analysis of herbivore-related WRKY genes in tea plants, and provided a good genetic resource and theoretical basis for future studies of pest-resistant genes and breeding of tea plants.

Study on the Differences in Ligand-binding Function and Mode of the Antennal High-abundance Odorant-binding Proteins EoblOBP9 and EoblOBP11 of the Tea Geometrid, Ectropis obliqua Prout

YAN Yuting, WU Fan, ZHANG Yali, FU Xiaobin, CUI Hongchun, HAN Baoyu, LI Hongliang

Journal of Tea Science. 2021, 41(5): 643-653. doi:10.13305/j.cnki.jts.2021.05.002

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As an important lepidopteran pest in tea gardens, the male and female adults of Ectropis obliqua Prout can perceive and distinguish the chemical information of the external environment through the olfactory system, which helps its behaviors such as mating and selection of oviposition locations. In this process, Odorant-binding proteins (OBPs) highly expressed in the antennae of adults may play an important role. In view of this, this study focused on the two OBPs high expressed in the antennae of the adult tea geometrid, namely EoblOBP9 and EoblOBP 11. First, it was found that both EoblOBP9 and EoblOBP11 contain six conserved cysteines, the typical characteristics of the OBPs family, while the isoelectric points are predicted to be acidic and basic, respectively, indicating their difference in the primary structural properties. Both recombinant proteins were obtained using prokaryotic expression technology, and the corresponding polyclonal antibodies were prepared by immunizing mice. It was verified that they were indeed expressed specifically in the antennae of male adults by Western blot. Their binding abilities with 20 candidate ligands were tested by fluorescence competitive binding experiments, and the results show that both of them were compatible with the two sex pheromone components (Z3,Z9-6,7-epo-18: Hy and Z3,Z6,Z9-18: Hy) and a plant volatile component (trans-2-hexenal) has a strong affinity. Meanwhile, two OBPs also showed different ligand binding spectra. For example, EoblOBP9 had a strong affinity with 1-penten-3-ol, benzyl alcohol, acetophenone and benzaldehyde, indicating that the binding profiles were wider; while EoblOBP11 had a strong affinity with α-terpineol with a more specific binding mode. Molecular docking analysis also supports this conclusion. In addition, it was predicted that the active sites of both proteins are located at the C-terminus, which also shows the commonality of ligand binding. In conclusion, this study shows that although both OBPs were specifically expressed in the adult tea geometrid with high abundance, they are both conservative and different in terms of structural properties and biochemical binding profiles. The general characters and significant differences are coexist, showing the high abundance and diversities of the OBPs. This can also explain an adaptation mechanism of the tea geometrid olfactory system in the face of the complex and changeable external environment.

Decomposition Characteristics of Lignin and Cellulose in Different Litters of Ecological Tea Gardens in Mountainous Areas of Guizhou

LIU Shaqian, YANG Rui, HOU Chunlan, MA Juebing, GUO Jiarui

Journal of Tea Science. 2021, 41(5): 654-668. doi:10.13305/j.cnki.jts.2021.05.003

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The litter decomposition bag method was used to analyze the decomposition of five single litter types and nine mixed litter types in Guizhou Jiu'an Ecological Tea Garden with Cinnamomum glanduliferum, Betula luminifera, Cunninghamia lanceolata and Pinus massoniana leaf litters and Camellia sinensis trims as the research objects. The decomposition rate and release characteristics and differences of lignin and cellulose were studied. The results show that in a single type of litter, the litter decomposition rate of broad-leaved species was higher than that of coniferous species. The lignin content of litter of broad-leaved species showed an increasing trend. While that of coniferous species showed a downward trend as a whole. The content of cellulose in litter also showed a decreasing trend. According to the analysis of the release law of lignin and cellulose, both single litter release law and mixed litter cellulose release law were direct release. There were three release laws for single litter lignin, and 5 laws for mixed litter lignin. According to the analysis of the decomposition characteristics of 9 kinds of mixed litter, 44.4% of mixed litter decomposition showed additive effect, 11.2% mixed decomposition showed antagonistic effect, and 44.4% mixed decomposition showed synergistic effect. While 22.2% of mixed litter lignin release showed promoting release, and 55.6% mixed litter lignin release promoted the enrichment. A total of 11.1% of the mixed litter cellulose appeared to promote the enrichment, and 22.2% of the mixed litter cellulose was to promote the release. This study provided a theoretical basis for further maintaining the fertility in tea gardens.

Preventive Effect of Green Brick Tea on Non-alcoholic Fatty Liver Disease via Gut Microbiota Changes

ZHOU Tingting, CHEN Guiting, CAO Nan, HE Jiangang, HE Gongwei, XIAO Changyi, LI Shigang

Journal of Tea Science. 2021, 41(5): 669-680. doi:10.13305/j.cnki.jts.20210914.001

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The preventive effect of green brick tea (GBT) on the mouse non-alcoholic fatty liver disease (NAFLD) model was studied by affecting changes in gut microbiota. C57BL/6 mice were randomly divided into 5 groups, including normal control group (NC), model control group (MC), positive drug control group (PC), low-dose group (LD) and high-dose group (HD) of GBT. A NAFLD model was established by feeding mice with high-fat diet, and supplemented with low and high doses of GBT water extract and positive drug (Xuezhikang) respectively. The body weight, food utilization efficiency, liver weight, liver index, TC, LDL-C/HDL-C and ALT contents of mice were determined. Liver tissue pathological sections were observed by HE staining and Oil Red O staining. ELISA method was used to detect changes of IL-1β and IL-18 in liver tissue. The changes of gut microbiota were analyzed by high-throughput sequencing in 16 S rDNA V3-V4 region, and Spearman's correlation analysis method was used to analyze the correlation between gut microbiota and NAFLD phenotype. Compared with the model control group, the body weight, food utilization efficiency, liver weight, liver index, serum TC, LDL-C/HDL-C, ALT, liver tissue TC, IL-1β, and IL-18 contents of mice in the GBT group were significantly reduced, and the degree of liver disease was improved. Gut microbiota analysis and correlation analysis show that the species abundance of Bacteroides decreased, and it was positively correlated with the NAFLD phenotype. The species abundance of Lactobacillus, Alloprevotella, and Saccharibacteria_genera_incertae_sedis increased, and they were negatively correlated with the NAFLD phenotype. Bacteroides and Lactobacillus had the strongest correlation with NAFLD phenotype. Green brick tea has a certain preventive effect on NAFLD, and its effect may be related to the changes in gut microbiota.

Effects of Fu Brick Tea on Inflammation and Intestinal Microflora Diversity in Mice with DSS-induced Ulcerative Colitis

HUANG Xiangxiang, TAN Ting, YU Lijun, WANG Kunbo, HUANG Jian'an, XU Shiyu, LIU Zhonghua

Journal of Tea Science. 2021, 41(5): 681-694. doi:10.13305/j.cnki.jts.2021.05.004

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In this study, we investigated the effects of Fu brick tea on the anti-inflammation and intestinal microflora of ulcerative colitis (UC) mice induced by Dextran sulfate sodium (DSS). Sixty C57BL/6 female mice were randomly divided into the normal control group, 10 mg·kg-1 EGCG control group, high dose brick tea (300 mg·kg-1, HBT) control group, DSS model group, DSS＋10 mg·kg-1 EGCG group, DSS＋low dose brick tea (100 mg·kg-1, LBT) group, DSS＋middle dose brick tea (150 mg·kg-1, MBT) group and DSS＋HBT group, with the control groups n=5 and the treatment groups n=9. After 1 week of DSS modeling, the mice were gavaged for 4 weeks. The mice were executed after 5 weeks of the treatments. The histopathological changes of mice colon were observed, and the disease activity index (DAI) of mice colitis was assessed. The levels of IL-6, IL-8, IL-1β, TNF-α and IFN-γ inflammatory factors in mice serum and the expression levels of JAK2 and STAT3 genes in mice colon tissues were measured. The intestinal microflora of mice was analyzed by PCR-DGGE and clone sequencing techniques. The results show that compared with the DSS model group, the quality of survival and colonic tissue morphology of mice were significantly improved after feeding EGCG and different concentrations of brick tea, and the levels of inflammatory factors IL-6, IL-8, IL-1β, TNF-α and IFN-γ in mice serum were significantly reduced (P<0.05). Moreover, the expressions of JAK2 and STAT3 genes were significantly reduced (P<0.05) JAK2/STAT3 inflammatory signaling pathway was inhibited; and intestinal microflora diversity and richness were increased significantly. The dominant bacterial flora in different treatment groups were changed as well. In conclusion, Fu brick tea can ameliorate DSS-induced UC injury by inhibiting JAK2/STAT3 inflammatory signaling pathway and modulating intestinal microflora diversity.

Dynamic Changes of Physiological Responses and Antioxidant Enzyme Activities in Tea Root System under Severe Shading and Removal of Shading

JIAO Haizhen, SHAO Chenyu, CHEN Jianjiao, ZHANG Chenyu, CHEN Jiahao, LI Yunfei, SHEN Chengwen

Journal of Tea Science. 2021, 41(5): 695-704. doi:10.13305/j.cnki.jts.2021.05.005

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Shading is one of the important measures to improve the quality of summer and autumn tea. There were a lot of studies on the effect of shading on tea leaves, but there were few reports on the root system. In this study, a pot experiment was used to explore the dynamic changes of physiological indexes and antioxidant enzyme activities of ‘Xiangfeicui’ and ‘Jinxuan’ under 95% shading treatment. Data were collected after shading for 0, 4, 12 d and the 4 d after regaining light. The results show that the number of new absorbing roots and lateral roots of tea plants increased after shading for 12 d, and decreased on the 4th day after regaining light. The trends of root vigor changes were the same. The soluble sugar content of ‘Xiangfeicui’ increased after shading, and the soluble sugars of ‘Xiangfeicui’ and ‘Jinxuan’ increased by 5.82% and 8.04% on the 4th day after regaining light. On the 4th day of heavy shading, the APX and POD activities of the two cultivars reached the peaks. On the 12th day of shading, the activities of SOD and APX the highest. The APX activity of tea roots after regaining light was significantly higher than that without shading. This study reveals that the roots of tea plants were osmotically regulated by increasing the content of soluble sugars, enhancing root vigor to absorb and transport nutrients, and increasing antioxidant enzyme activities to scavenging free radicals in response to heavy shade environments. Under a shading environment (95%), ‘Jinxuan’ was more shade-tolerant, and regaining light might cause a certain degree of stress to the roots of tea plants.

Design of End Effector for Picking Tea Shoots

XU Lijia, LIU Qi, DAI Jianwu, CHEN Xiaohu

Journal of Tea Science. 2021, 41(5): 705-716. doi:10.13305/j.cnki.jts.2021.05.006

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In order to solve the selective picking of tea shoots, based on the physical characteristics of tea shoots, an end effector of picking tea shoots with clip was designed in this study. The GUI module of MATLAB software and SolidWorks software were used to optimize the structural parameters of the picking end effector,and the optimal combination structural parameters were obtained. The kinematics simulation analysis was feasible and met the requirements of clamping force. The model of picking end effector was tested in tea garden, and the results show that the missed picking rate of one bud and one leaf was 2.8%, and the picking integrity rate was 91%. While the missed picking rate of one bud and two leaves was <3%, and the picking integrity rate was about 94% in March. The experimental results verified that the structural design and parameters of the picking end effector are feasible, which provided a theoretical basis for the selective picking of tea shoots, and a technical support for the subsequent research and development of tea shoot picking machine.

Rapid Determination of 24 Pesticide Residues in Tea by in-Syringe Dispersive Solid Phase Extraction-Ultra High Performance Liquid Chromatography-Tandem Mass Spectrometry

MA Jiali, WANG Chen, CHEN Hongping, CHAI Yunfeng, ZHU Li, LIU Xin

Journal of Tea Science. 2021, 41(5): 717-730. doi:10.13305/j.cnki.jts.20210804.001

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Based on the characteristics of tea matrix, a rapid pretreatment technology of in-syringe dispersive solid phase extraction (dSPE) was developed, and an analytical method based on ultra high performance liquid chromatography-tandem mass spectrometry was established for the detection of 24 pesticide residues in tea. Tea samples were extracted using acetonitrile, salted-out with MgSO₄, purified with primary secondary amine and graphitized carbon black as dispersive adsorbents in designed syringe devices, and detected by UPLC-MS/MS. Recoveries of 24 pesticides in black and green teas were 61.7%-98.8% at three spiked levels (0.01βmg·kg^-1, 0.05βmg·kg^-1 and 0.5βmg·kg^-1), which indicated high accuracy with the relative standard deviations of 0.4%-5.5%. The calibration curves of 24 pesticides in black and green teas showed good linearity with correlation coefficients (R²) higher than 0.995. The limits of detection (LOD) and the limits of quantitation (LOQ) were 0.05-5.36βμg·kg^-1 and 0.18-17.86βμg·kg^-1, respectively, which show high sensitivity of this method. Being simple, fast, less instrument required and time-saving, this method is suitable for quantification of multiple pesticide residues in tea.

Study on the Effect Measurement of the Integration Development of Tea Industry and Tourism Industry in Yangtze River Delta

WEI Nixi, XIANG Guopeng

Journal of Tea Science. 2021, 41(5): 731-742. doi:10.13305/j.cnki.jts.20210804.003

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Based on the provincial panel data from 2011 to 2019, the integration effect of tea industry and tourism industry in the Yangtze River Delta was measured by the coupling coordination degree model. The results show that the comprehensive development level of tea industry and tourism industry in the Yangtze River Delta had been continuously improved, but there were some differences between the two industries. The staged and regional imbalance of tea industry development was prominent, while the regional synergy effect of tourism industry was significant. The synchronous characteristics of tea industry and tourism industry in the Yangtze River Delta were obvious, and the trend of industrial convergence was obvious. However, the rising trend of coupling coordination degree was slow. As a whole, there was a long way to go to promote the benign interaction and high-quality integration development of tea tourism industry system in the Yangtze River Delta. Finally, the countermeasures for the integration of tea industry and tourism industry in Yangtze River Delta were put forward.

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