Journal of Tea Science

Effects of Intercropping Functional Plants on the Ecosystem Functions and Services in Tea Garden

SHI Fan, HUANG Hongjing, CHEN Yanting, CHEN Lilin

Journal of Tea Science. 2022, 42(2): 151-168. doi:10.13305/j.cnki.jts.2022.02.011

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As one of the main measures of habitat management, reasonable intercropping of functional plants in tea garden can shade tea bushes and keep them warm, conserve water and soil, increase fertility and promote growth, as well as maintain micro-habitat stability. It also can attract natural enemies, repel pests, reduce the damage caused by diseases, insects, and weeds in tea garden, therefore it is beneficial for improving the quality and efficiency of the tea. However, unreasonable intercropping will destroy the micro-habitat of tea garden, compete for nutrients, thus affecting the growth of tea plants. In this paper, the intercropping of functional plants and their effects on the ecosystem functions and services in tea garden in recent years were reviewed. These effects mainly included the species, management models, functions and common problems with the most widely used functional plants, as well as the regulatory effects of intercropping of functional plants on the growth and development of tea plants, tea quality and yield, and tea pests. The ultimate goal of this paper was to provide guidance for the application of intercropping measures to promote the comprehensive regulation of pests in tea garden, and enhance the ecosystem functions and services of tea garden.

Research Progress of Tea Beer

CHEN Dequan, ZHU Yan, ZOU Chun, YIN Junfeng, CHEN Jianxin, XU Yongquan

Journal of Tea Science. 2022, 42(2): 169-178. doi:10.13305/j.cnki.jts.2022.02.005

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Tea beer is a new type of beer, obtained by adding tea or tea extract in the brewing process with the dual flavor characteristics of tea and beer. The development of tea beer would not only enrich the types, flavors, and physiological effects of beer, but also improve the utilization rate and additional value of tea resources and benefit for the common development of tea and beer industries. However, the studies of tea beer on pretreatment of raw materials, fermentation and clarification technologies were still not mature. Therefore, this paper systemically summarized the processing technologies of tea beer, including the pretreatment of raw materials, fermentation and clarification technologies. Based on that, we made the prospects for the future of tea beer, aiming to provide a reference for the research and development of tea beer in depth.

Screening and Identification of Chaperone CsHIPP26.1 Chelating Ions in Tea Cultivar ‘Huangjinya’

LIU Fuhao, FAN Yangen, WANG Yu, MENG Fanyue, ZHANG Lixia

Journal of Tea Science. 2022, 42(2): 179-186. doi:10.13305/j.cnki.jts.2022.02.006

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Heavy metal-associated isoprenylated plant proteins (HIPPs) is an important metallochaperones due to its unique heavy metal binding domains (HMA) and the structural characteristics of isoprenylation motif. In order to identify the chelating ions of CsHIPP26.1 protein in Camellia sinensis (L.) cv. ‘Huangjinya’, the pET-32a-CsHIPP26.1 recombinant plasmids and empty carriers were respectively transferred into E. coli BL21, and then were cultured in LB liquid culture medium with 4 mol·L^-1 single metal ions (CuCl₂, ZnCl₂, MgCl₂, FeCl₃, CaCl₂) or multiple metal ions and 1 mmol·L^-1 IPTG. The growth of E. coli in different ion media was observed, meanwhile the fusion target protein was obtained by His-tag protein purification magnetic bead. The contents of metal ions in fusion protein were analyzed by atomic absorption spectrophotometer, and the number of ions chelated by the protein was calculated. The results show that CsHIPP26.1 protein was only chelated with Zn²⁺ and Cu²⁺, and the chelating ability to Zn²⁺ was significantly higher than Cu²⁺. Based on the molar ratio of its bound metal ions to the target protein, the maximum number of Zn²⁺, Cu²⁺ chelated by CsHIPP26.1 protein was 2 and 1, respectively.

Effects of Exogenous 5-ALA on the Chlorophyll Synthesis and Fluorescence Characteristics and Gene Expression of Key Enzymes in Tea Plants under Drought Stress

YANG Ni, LI Yimin, Li Jingwen, TENG Ruimin, CHEN Yi, WANG Yahui, ZHUANG Jing

Journal of Tea Science. 2022, 42(2): 187-199. doi:10.13305/j.cnki.jts.2022.02.010

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In order to study the regulation mechanism of exogenous 5-aminolevulinic acid (5-ALA) on the chlorophyll synthesis and fluorescence characteristics of tea plants in response to drought stress, tea cultivar 'Shuchazao' was used as the experimental material in this study. Under PEG-6000 simulated drought stress environment and 5-ALA spraying pretreatment, the contents of chlorophyll a, chlorophyll b and total chlorophyll in tea seedling leaves were detected, and the chlorophyll fluorescence parameters and the expressions of key enzyme genes in tea leaves were further determined. The results show that exogenous 5-ALA significantly increased the contents of chlorophyll a, chlorophyll b and total chlorophyll in tea leaves under drought stress. Exogenous 5-ALA alleviated the decreases of maximum fluorescence (Fm), actual photochemical efficiency of PSⅡ [Y(Ⅱ)], maximum photochemical quantum yield of PSⅡ (Fv/Fm), photochemical quenching coefficient (qP), potential activity of PSⅡ(Fv/Fo), photosynthetic electron transfer rate (ETR) of PSⅡ reaction center, and the increases of initial fluorescence (Fo) and nonphotochemical quenching coefficient (qN). At the same time, exogenous 5-ALA induced the expressions of chlorophyll synthesis (CsHEMA1, CsHEME1, CsLIN2) and carbon assimilation-related enzymes (CsSBPase, CsTK) in tea plant under drought stress. The study demonstrates that spraying exogenous 5-ALA can effectively alleviate the degradation of chlorophyll and damage to PSⅡ reaction center of tea leaves under drought stress, maintain the higher photosynthetic activity, and improve the photoprotection ability of tea leaves.

The Expression Profiles of Chemosensory Protein 8 Orthologs in Two Closely Related Tea Geometrid Species, Ectropis obliqua Prout and Ectropis grisescens Warren

YAN Yuting, LI Yujie, WANG Qian, TANG Meijun, GUO Huawei, LI Hongliang, SUN Liang

Journal of Tea Science. 2022, 42(2): 200-210. doi:10.13305/j.cnki.jts.2022.02.007

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Chemosensory proteins (CSPs) play crucial roles in insect chemosensory and non-chemosensory processes. Ectropis obliqua Prout and its sibling species, Ectropis grisescens Warren are serious lepidopteran moth pests in tea gardens and they cause destructive damages to tea plants. The development of greener and environmentally friendly pest managements would benefit from orthologous CSPs’ investigation on the interaction between two Ectropis species and tea plants. This study mainly examined the expression profiles of EgriCSP8, the orthologs of EoblCSP8 in E. grisescens by using the real-time quantitative PCR experiment. The qRT-PCR results show that EgriCSP8 and EoblCSP8 had a conserved larvae-enriched expression pattern. EgriCSP8 was mainly expressed in the heads of the third instar of E. grisescens larvae. Next, the expression levels of CSP8 were compared between E. obliqua and E. grisescens. The results show that the expression level of EoblCSP8 in E. obliqua was higher than that of EgriCSP8 in E. grisescens. In addition, the effects of Wolbachia and tea plant volatiles on CSP8 expression levels in E. obliqua and E. grisescens were also estimated. The results show that the EgriCSP8 expression level declined significantly when Wolbachia was removed in E. grisescens, which provided the first evidence that the insect CSPs’ expression is associated with Wolbachia. These findings laid a foundation for the future studies on the molecular mechanisms of chemosensory and coevolution between the geometrid sibling species and tea plants.

The Screening, Identification and Enzyme Production of Thermophilic Bacteria in Pile-fermentation of Qingzhuan Tea

ZHU Wen, WU Shuang, WANG Wenfeng, XU Wencan, CHEN Wenjun, HUANG Youyi

Journal of Tea Science. 2022, 42(2): 211-221. doi:10.13305/j.cnki.jts.2022.02.001

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Pile-fermentation (PF) is a key step for the formation of the unique quality of Qingzhuan tea. In this study, high-temperature screening and identification of bacteria from PF samples of Qingzhuan tea were carried out. The results show that 20 strains of bacteria that could grow under high temperature conditions were obtained through screening, of which 15 strains could grow on a tea-containing medium. The optimum temperature of 3 thermophilic bacteria strains was 55℃. Combined with the morphological characteristics and the 16 S rDNA gene sequence analysis, it was found that one strain was Bacillus subtilis and the other two were Bacillus licheniformis. The enzyme production test on raw tea by thermophilic bacteria shows that three thermophilic bacteria could produce cellulase, amylase and tannin during tea fermentation, and the enzyme activities reached 215.69 U, 259.28 U, and 4.85 U, respectively.

Effects of Anaerobic Treatment on Amino Acid Composition and Biological Activities of Different Type Teas

YANG Gaozhong, PENG Qunhua, ZHANG Yue, SHI Jiang, LIN Zhi, LÜ Haipeng

Journal of Tea Science. 2022, 42(2): 222-232. doi:10.13305/j.cnki.jts.2022.02.002

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The same batch of fresh tea leaves after 7 hours anaerobic treatment was processed into green tea, white tea, oolong tea, black tea, and dark raw tea according to different processing technologies. The amino acid composition and content, angiotensin converting enzyme (ACE) inhibitory activity were determined, and their antioxidant activities were also measured by determining cellular antioxidant activity (CAA), 1,1-diphenyl-2-picrylhydrazyl (DPPH) free radical scavenging capacity, 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) cation radical scavenging capacity, superoxide anion radical absorbance capacity, and ferric ion reducing antioxidant power (FRAP). Results show that the composition and content of free amino acids in different types of teas varied significantly which were processed after anaerobic treatment, and especially the content of γ-aminobutyric acid (GABA) increased significantly. The highest GABA content was found in green tea (2.21 mg·g^-1), followed by oolong tea (2.19 mg·g^-1), while the lowest in black tea (0.86 mg·g^-1). Moreover, green tea and dark raw tea showed the strongest inhibition rate of ACE (35.7% and 37.2%, respectively), while black tea showed the weakest inhibitory activity against ACE (23.8%). Additionally, it was found that green tea had the strongest antioxidant activity among the different types of teas processed from the same fresh tea leaves after anaerobic treatment.

Variation of Oil Content in Tea Seed Kernel and Fatty Acid Compositions and Contents in Tea Seed Oil among Different Culitivars and Regions

XIANG Jing, LIANG Yuerong, ZHAO Dong, WANG Kairong, LU Jianliang, YUAN Ming'an, ZHENG Xinqiang

Journal of Tea Science. 2022, 42(2): 233-248. doi:10.13305/j.cnki.jts.2022.02.008

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In order to identify tea cultivars with high oil and rich fatty acid compositions, the relevant research on tea seeds of 49 tea cultivars from 9 provinces was carried out. The results show that the oil content of different cultivars were 16.29%-33.80%. The composition and content of fatty acids in tea seed oil were determined by gas chromatography. A total of 19 kinds of fatty acids were detected, among which oleic acid, linoleic acid, palmitic acid, stearic acid and α-linolenic acid were the main fatty acids accounting for 46.00%-72.64%, 8.05%-31.05%, 12.02%-18.80%, 2.16%-5.34% and 0.36%-1.20% of the total fatty acids content, respectively. The average ratio of saturated fatty acid, monounsaturated fatty acid and polyunsaturated fatty acid content in different cultivars of tea seed oil was generally 1∶3.04∶1.40. The correlation analysis of fatty acid components shows that cis-oleic acid was negatively correlated with trans-oleic acid and docosahexaenoic acid, while cis-oleic acid was positive correlated with α-linolenic acid and palmitic acid, and trans-oleic acid was negatively correlated with palmitic acid. There were significant differences in fatty acid content in different regions. Based on the present results, ‘Zhongcha 108’ from Zhejiang province, ‘Jinxuan’ from Guangdong province, ‘Xiangbolv 2’ and ‘Zhuyeqi’ could be used as good oil tea cultivars since their seeds have higher fatty acid contents and the proportion of three fatty acids types was more balanced. ‘Baoshan population’, ‘Xiangshanzao’, ‘Jinxuan’ from Hunan province, ‘Zijuan’ and ‘Yunnan wild tea population’ had high unsaturated fatty acid contents, which were suitable for making pharmaceutical and cosmetic oil.

Determination of Multi-pesticide Residues in Tea Based on the Combination of CS/GO/DM SPE Column and UPLC-MS/MS

HU Gaohua, CAO Jianrong, YANG-LEI Wenxuan, WANG Chen, ZHOU Sujuan, LIU Xin, LU Chengyin, CHEN Hongping, MA Guicen

Journal of Tea Science. 2022, 42(2): 249-262. doi:10.13305/j.cnki.jts.2022.02.004

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In this study, the synthesized chitosan/graphene oxide/diatomite (CS/GO/DM) solid phase extraction (SPE) column cartridge was used for the purification of tea matrix. A method for determination of 46 kinds of pesticide residues in tea by using ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was established. The tea samples were extracted by acetonitrile, purified by CS/GO/DM SPE column, washed by acetonitrile. Then, the target compounds of 46 pesticide residues were analyzed by UPLC-MS/MS. The results show that CS/GO/DM SPE column could efficiently remove tea interference substances, which could greatly reduce the matrix effect of pesticides in green tea, black tea and oolong tea by 4.7%-66.5%, 3.2%-35.5% and 4.4%-42.8%, respectively. The recoveries of 46 kinds of pesticides ranged from 61.5% to 118% at three spiked levels, with good linearity. The correlation coefficients (R²) were all higher than 0.98. The developed method has the great advantages of operation simplicity, time saving, and high cleanup ability with only one kind towards different types of tea, no need for using matrix standard solution and a low amount of adsorbent.

Development and Test for Distributed Control Prototype of the Riding Profiling Tea Harvester

ZHAO Runmao, BIAN Xianbing, CHEN Jianneng, DONG Chunwang, WU Chuanyu, JIA Jiangming, MAO Ming, XIONG Yongsen

Journal of Tea Science. 2022, 42(2): 263-276. doi:10.13305/j.cnki.jts.2022.02.003

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To better realize the mechanized picking of ordinary fresh tea leaves, reduce the system development cost, and improve the automation level and picking integrity rate, a profiling method of machine-made tea canopy surface cutter and a development method of low-cost and rapid control prototype machine with distributed control were both put forward in this study. Additionally, the profiling tea harvester prototype was developed and the indoor and field tests were launched. The profiling tea harvester uses ultrasonic sensor to sense the distance information from the reciprocating cutter to the tea canopy and realizes profiling transmission through screw rods. Then, Hampel filter and Low-pass filter algorithms are used to preprocess the distance information on-line to eliminate the influence of inter leaf vacancy and cutter vibration on the real height estimation of tea canopy surface. Meanwhile, a proportional nonlinear tracking differentiator (PNTD) control system based on nonlinear tracking differentiator was established in order to avoid the amplification of signal noise by digital difference in conventional PD control and improve the accuracy of profiling of each tea picking unit. Based on CAN bus network and code generation technology, the distributed fast control prototype with multiple execution units was realized and tested. The indoor step test results show that the maximum peak time of this control system was 0.14 s and the maximum overshoot was 3.6%. Finally, to verify the effectiveness of the distributed control profiling tea picking prototype, profiling picking experiments were conducted in the tea garden. The average picking rate of young shoots on the canopy covered by cutter was 92.01%,the integrity rate of buds and leaves was 82.6%, the impurity rate was 6.4%, and the young shoots with at least one bud and three leaves accounted for 87.91%. All the results demonstrate that the proposed methods had good performance and could be used for picking ordinary tea.

Responses of Different Tea Cultivars (Strains) to Half Fertilization

SHU Zaifa, ZHENG Shenghong, SHAO Jingna, ZHOU Huijuan, JI Qingyong, LIU Yu, HE Weizhong, WANG Liyuan

Journal of Tea Science. 2022, 42(2): 277-289. doi:10.13305/j.cnki.jts.2022.02.009

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In this study, a field experiment was conducted to compare five tea cultivars (strains) under half fertilization and conventional fertilization for two years. The response of half fertilization to tea sprouting period, yield and quality was studied. The results show that compared with the conventional fertilization, half fertilization had little effect on the sprouting period of five tea cultivars (strains). Under half fertilization, the tea yield, free amino acid, caffeine and theanine contents decreased, while the content of polyphenols, ester catechin, non-ester catechin and phenol-ammonia ratio increased. The sensory evaluation results were consistent with the chemical component changes in the tea. Compared with the conventional fertilization, the average yield reduction of Zhongming 6, Zhongming 7, Zhongming 192, Wanghai Tea 1 and Longjing 43 under half fertilization were 4.19%, 12.09%, 2.13%, 22.59% and 3.29% respectively. Under half fertilization, Wanghai Tea 1 had an obvious response, Longjing 43 and Zhongming 7 had relatively significant responses, and Zhongming 6 and Zhongming 192 had little response. Except the yield of Wanghai Tea 1 decreased significantly, the tea yield and quality show no obvious change in the rest tea cultivars (strains) under different fertilization treatments. To a certain extent, the results indicate that half fertilization would not significantly affect the benefit of tea gardens within 2 years. The results also provided a preliminary theoretical support for the implementation of weight loss and stability of tea garden benefits.

Research on the Changes of China's Tea Production Layout

WU Qinyao, YANG Jiangfan, LIN Cheng, GUAN Xi

Journal of Tea Science. 2022, 42(2): 290-300. doi:10.13305/j.cnki.jts.20211210.001

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Based on the agricultural location theory, comparative advantage theory and rational "economic man" hypothesis, a theoretical analysis framework taking cultural factors and factors affecting the changes in China's tea production layout into account was constructed using data from 16 tea-producing provinces across the country from 1993 to 2018. The panel data of the district expounded the characteristics and laws of the changes in China's tea production layout, and clarified the main influencing factors and driving mechanisms that affected the changes in China's tea production layout. The results show that: (1) After the reform and opening up, from 1993 to 2018, China's tea industry had agglomerated and expanded in terms of production scale, and the tea production space had changed significantly, with significant differences in characteristics. (2) The layout of tea yield was based on natural resources, consumer demand, and culture. The factors like technological progress and social and economic factors had a profound impact on the layout of tea production over time, and resulted in differences in tea-producing provinces across the country. (3) Natural resources and technological progress were the basis and prerequisite for changes in the layout of tea production. The decisive conditions for changes in the layout of tea production were consumer demand and socio-economic factors. Brand building and policy support were also important supplementary factors for the current development of the tea industry. On this basis, this article proposed to optimize the layout of tea production according to local conditions, strengthen the protection of land resources, rationally plan the area of tea gardens, maintain the continuous and stable development of the tea industry, attach importance to technological innovation and application to improve the level of tea yields and use the inter-regional interaction of tea production to achieve policy recommendations such as improving the overall efficiency of the tea industry.

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