Journal of Tea Science

Recent Advance on Anti-cardiovascular Inflammation of Major Characteristic Compounds in Tea

YAO Min, LI Daxiang, XIE Zhongwen

Journal of Tea Science. 2020, 40(1): 1-14. doi:10.13305/j.cnki.jts.20191202.004

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Chronic inflammation is one of the important causes of cardiovascular diseases such as atherosclerosis, hypertension and myocardial infarction. Inflammatory factors such as IL-6, TNF-α and IL-1β can induce many cardiovascular diseases, including myocardial hypertrophy, myocardial fibrosis and abnormal proliferation of vascular smooth muscle cells. Tea is rich in characteristic compounds. These compounds play preventive and therapeutic roles for cardiovascular diseases by inhibiting proinflammatory cytokines production, and regulating the signal pathways such as NF-κB, MAPK and Tolls-like receptors. In this review, we summarized the recent advance of the main characteristic compounds in tea on preventing inflammation related cardiovascular diseases.

Study and Prospect of Soil Acidification Causes and Improvement Measures in Tea Plantation

FAN Zhanhui, TANG Xiaojun, ZHENG Dan, YANG Qin, CHEN Guangnian, LI Xiaowen, SUN Jiabin

Journal of Tea Science. 2020, 40(1): 15-25. doi:10.13305/j.cnki.jts.20200117.006

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In recent years, the acidification of tea garden soil has become increasingly serious due to the influences of internal and external factors. Soil acidification of tea plantation mainly manifests as soil structural deterioration, reduced fertility and increased heavy metal content. These variations can affect the growth and development of tea trees and reduce the tea quality, and may even harm human health, seriously threaten the sustainable development of tea plantation. This paper reviewed the research advance on soil acidification of tea plantations in recent years, summarized the causes, harms and improvement measures for soil acidification, and finally proposed the future research directions.

cDNA-AFLP Reveals Differential Gene Expression Profiles of Tea Plant (Camellia sinensis cv. Maoxie) Induced by Colletotrichum sp.1 Infection

WEI Rifeng, LAI Jiandong, PENG Chengbin, ZHANG Chengkang, LIAN Lingli, LIU Wei

Journal of Tea Science. 2020, 40(1): 26-38. doi:10.13305/j.cnki.jts.20200117.001

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Differentially expressed genes related to Camellia sinensis cv. Maoxie against Colletotrichum sp.1 infection were screened by cDNA-AFLP (cDNA-amplified fragment length polymorphism) to provide reference for further illustrating the molecular mechanism underlying tea resistance to anthracnose. A total of 256 pairs of selective primers were applied to amplify the cDNA of leaves collected at 0 h and 48 h after Colletotrichum infection, and 136 differentially expressed transcript-derived fragments (TDFs) were obtained via screening, sequencing and BLAST analysis. The analysis of the homologous genes revealed that 128 TDFs were homologous with the known genes in Nr database. Most of them were related to stress responses, biological regulation and signal transduction, and 51 TDFs were highly homologous to differential expressed genes of tea plant under cold, drought or salt conditions in TPIA database. Further qRT-PCR analysis of 27 TDFs showed that the expression profiles of 24 TDFs were consistent with those of cDNA-AFLP results, among which several resistance-related genes including WRKY transcription factor, ethylene-responsive transcription factor, peroxidase and superoxide dismutase were significantly up-regulated. The differential expressed genes found in this study would lay a foundation for further analysis of their functions.

Cloning and Expression Analysis of CsPPH Gene in Tea Plant (Camellia sinensis)

ZHOU Zhe, CHEN Zhidan, WU Quanjin, XU Yilan, SUN Weijiang

Journal of Tea Science. 2020, 40(1): 39-50. doi:10.13305/j.cnki.jts.20200117.007

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Pheophytinase (PPH) is a key enzyme in chlorophyll degradation. It can convert pheophytin a into pheophorbide a, which is the last product to keep green in chlorophyll degradation pathway. This step is considered to be a key step of leaf senescence and yellowing. In this study, the full-length sequence of CsPPH gene was cloned from the new shoot leaves of albino tea plant Camellia sinensis cv. Baijiguan (MK359094), and its biological characteristics were analyzed. The full-length of CsPPH was 1 298 bp, and the ORF was 1 241 bp, encoding 413 amino acids. Sequence analysis indicated that the encoded protein was a stable hydrophobic protein, and its molecular weight was predicted to be 45 741.50 Da. Its theoretical isoelectric point was 6.12. It was mainly located in chloroplasts. The real-time fluorescence quantitative PCR showed that under shading conditions, the expression of CsPPH was inhibited, chlorophyll increased and leaf color turned green. Light promoted the expression of CsPPH in leaves of cv. Baijiguan, and chlorophyll degradation led to leaf albinism.

Optimization of Technical Parameters for Chemical Synthesis of Theasinensin A by PBD and RSM

ZHANG Jianyong, CHEN Lin, CUI Hongchun, WANG Weiwei, XUE Jinjin, XIONG Chunhua, JIANG Heyuan

Journal of Tea Science. 2020, 40(1): 51-62. doi:10.13305/j.cnki.jts.20200117.002

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The effect of chemical synthesis conditions on the yield of theasinensin A was studied. Key factors for chemical synthesis of theasinensin A (TSA) were determined by single factor test and Plackett-Burman Design (PBD). The response surface methodology (RSM) was employed to optimize the technical parameters for the chemical synthesis. The experiment results indicate that the effects of copper oxide content, methanol content and temperature on the yield were extremely significant. The main factors were methanol content, next copper oxide content, and then temperature. The optimal conditions were as follows: copper chloride content 43%, methanol content 26%, and temperature 15℃. Under these conditions, the yield of TSA was 59.12%, which was close to the predicted value of 59.34%. The combination of PBD and RSM to optimize the chemical synthesis of TSA achieved ideal result, which could provide reference and theoretical basis for the efficient chemical synthesis of other catechin oxidation polymers.

Leaching Pattern of Internal Substances and Xenobiotic Pollutants during Tea Brewing

CHEN Hongping, LIU Xin, LU Chengyin, QIU Jing

Journal of Tea Science. 2020, 40(1): 63-76. doi:10.13305/j.cnki.jts.20200117.008

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Based on the recent literatures, the dissolving, releasing and transformation patterns of tea internal compounds and exogenous contaminants were summarized and discussed in this study. Physicochemical properties of chemicals and tea brewing methods are the internal and external cause affecting extraction rates of chemicals during tea brewing. Internal cause plays a crucial role in extraction rates, while external cause is more prominent for the leaching of tea internal substances. Water solubility of chemicals is positively correlated with extraction rates, while octanol-water partition is negatively correlated with extraction rates. Increasing water temperature is helpful for increasing extraction rates of chemicals and their concentrations in tea infusion. Brewing time is negatively correlated with extraction rates in a period of time, while the concentrations of chemicals in tea infusion increase with the brewing time. Compared with other pesticides, most of neonicotine pesticides and carbamate pesticides have higher extraction rates over 60%. The results of extraction rates of metal elements in tea infusion are quite different, and extraction rates of fluorine, nickel and cobalt have high extraction rates over 50%, while extraction rates of lead range from 20% to 50%. Metabonomic analysis based-high resolution mass spectrometry technique is a promising and powerful method for profiling extracting pattern of chemicals during tea brewing. Meanwhile, extracting behavior of toxic compounds during tea brewing will be deeply understood by using thermodynamic theory and kinetic model of mass extraction.

Absorption and Accumulation Characteristics of Selenium in Tea Plant (Camellia sinensis) and Expression Analysis of Genes Related to Selenium Regulation

CAO Dan, MA Linlong, LIU Yanli, GONG Ziming, JIN Xiaofang

Journal of Tea Science. 2020, 40(1): 77-84. doi:10.13305/j.cnki.jts.20191202.002

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The concentration dependence and time course of selenium (Se) absorption and accumulation in tea plant were investigated under sand culture and the expressions of related genes were also analyzed. The results show that the absorption amounts of Se in different tissues of tea plant were remarkably different. Most of them were fixed by the roots, and low movement from roots to shoots was observed. Moreover, the accumulation of Se was significantly correlated with exogenous Se concentration and culture duration. Furthermore, tea plant grew well in the Se concentration from 0 to 0.05 mmol·L^-1, but when the concentration was higher than 0.10 mmol·L^-1, tea plant showed poisoning symptoms. Fluorescence quantitative PCR analysis indicates that allene oxide cyclase (CsAOC), lipoxygenase (CsLOX6) (key genes of jasmonic acid signaling pathway), as well as a basic helix-loop-helix transcription factor (CsbHLH62) could be obviously induced by high concentration of Se. The correlation analysis showes that the expressions of these genes were positively related with the Se content in roots. These results suggest that Se accumulation in different tissues of tea plant was significantly correlated with exogenous Se concentration and culture duration, and CsbHLH62, CsAOC, CsLOX6 might play important roles in this process.

Integrated Nutrient Management in Tea Plantation to Reduce Chemical Fertilizer and Increase Nutrient Use Efficiency

RUAN Jianyun, MA Lifeng, YI Xiaoyun, SHI Yuanzhi, NI Kang, LIU Meiya, ZHANG Qunfeng

Journal of Tea Science. 2020, 40(1): 85-95. doi:10.13305/j.cnki.jts.20200117.003

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Tea is an important agricultural industry with great comparative advantage in southern China. However, there are some problems in tea fertilization inducing large nutrient loss, higher production cost and environmental risks. These problems are mainly excessive nutrient input, low proportion of specialized compound fertilizer for tea, low substitution rate of organic nutrients and low efficient application methods such as surface broadcasting. This paper summarized previous results and formulated a technical strategy of integrated nutrient management in tea plantation (INMT) including optimizing nutrient input amounts, properly replacing part of chemical fertilizer with organic fertilizers, choosing right fertilizer products, improving fertilization methods and ameliorating soil properties. Optimization of nutrient input amounts was achieved by controlling whole annual N input on the basis of tea type and yield level, and P and K on their relevant ratio to N and soil test if available. Limits of top input for these nutrients were suggested. Reasonable replacement ratio of organic fertilizer was recommended at 25%-50% of the total N input. A compound fertilizer with specialized formula for tea was developed and tested in more than 70 plantations across the main tea producing areas, efficiently increasing yield and quality while saving nutrient input. Fertilizer application was improved through changing topdressing period and adopting deep and mechanical application.Integrated nutrient management of tea was evaluated in 9 farmers’ and cooperative tea plantations. The average yield was increased by 9% in contrast with 42% reduction of average nutrient input. The tea quality was maintained or improved as indicated by slightly increased contents of amino acids and tea polyphenols.

Effects of Fertigation on Young Tea Plant Growth and Nutrient Absorption

YANG Qinglin, YANG Xiangde, JI Lingfei, MA Lifeng, RUAN Jianyun

Journal of Tea Science. 2020, 40(1): 96-104. doi:10.13305/j.cnki.jts.20200117.009

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Through field experiment, the effects of fertigation on the growth, nutrient absorption and quality components of young tea plants of Longjing43 and Baiye 1 were studied. We also compared the input costs under different fertilization patterns. The results show that the tea plant growth including tree height, trunk diameter and canopy width increased significantly under fertigation, even if the chemical fertilizer was reduced by 25%, compared with conventional furrow fertilization, and pruning biomass and nutrient uptake also increased significantly. At the same time, nutrient leakage loss was significantly reduced and nutrient use efficiency was improved. From the analysis of the times of fertigation, it could achieve better results by applying fertilizer 7 times per year under the experimental conditions for Longjing 43, and 14 times per year for Baiye 1.

The Fall Armyworm Spodoptera frugiperda may Transfer to Damage Tea Plant (Camellia sinesis)

SUN Xiaoling, CHEN Chengcong, LI Ning, LIU Fengjing, DONG Ya′nan, QIAN Xiaona, XING Yuxian, LIU Miaomiao, LI Xiwang

Journal of Tea Science. 2020, 40(1): 105-112. doi:10.13305/j.cnki.jts.20191202.001

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Spodoptera frugiperda is a world-class major migratory pest with a wide range of hosts. The growth fitness of S. frugiperda larvae feeding on maize and tea leaves was studied. The result shows that the mortality rate of the newly emerged larvae feeding on tea leaves was as high as 100% for 8 d. Weight gain of 3^rd instar larvae feeding on maize leaves was significantly higher than that of larvae feeding on tea leaves. After feeding for 3 d and 6 d, the weight gains of larvae feeding on tea leaves were 35.58% and 36.67% of those feeding on maize leaves respectively. Growth fitness of larvae feeding on maize leaves was significantly better than larvae feeding on tea leaves. The main manifestations are as follows: (1) the survival rate of the old larvae feeding on the maize leaves for 8 d was 20% higher than that feeding on tea leaves. (2) the pupation rate of 9-daysinstar larvae feeding on tea leaves was 54.17%. (3) the developmental period of the larvae feeding on tea leaves was significantly longer than that of the larvae feeding on the maize leaves. moreover, the pupa weight, length and width of the larvae feeding on maize leaves were significantly higher than those of the larvae feeding on tea leaves. Although the performance of S. frugiperda feeding on tea leaves was significantly lower than that feeding on maize leaves, some weeds species are the host of S. frugiperda and 3^rd instar larvae feeding on tea leaves could pupate successfully. Thus it is reasonable to speculate that if S. frugiperda migrates to the tea plantation or nearby, the larvae may transfer to damage tea plants with some weeds acting as intermediate.

Transcriptome Analysis of the Tea Leaves (Camellia sinensis var. assamica) Infected by Tea Blister Blight

SUN Yunnan, XU Yan, RAN Longxun, JIANG Huibing, SONG Weixi, XIA Lifei, CHEN Linbo, LIANG Mingzhi

Journal of Tea Science. 2020, 40(1): 113-124. doi:10.13305/j.cnki.jts.2020.01.001

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Illumina HiSeq2500, a high-through transcriptome sequencing technology, was applied for transcriptome analysis of tea leaves infected by tea blister blight. Through differential expression analysis, a total of 359 differentially expressed genes (DEGs）were identified after infection, of which 248 were up-regulated and 111 were down-regulated. With GO function annotation classifications, a total of 216 genes were divided into 122 function categories. The mainly involved functional categories included biological synthesis process, catalytic activity, cell process and many other physiological and biochemical processes. KEGG enrichment analysis showed that a total of 106 genes were annotated to 47 metabolic pathways, with monoterpenoid biosynthesis, porphyrin and chlorophyll metabolism, ribosome, nitrogen metabolism, diterpenoid biosynthesis, plant-pathogen interaction pathway significantly enriched. There were 32 differentially expressed transcription factors (TFs). Those TFs were classified into 16 families. qRT-PCR of randomly selected differentially expressed genes was used to validate transcriptome result, which showed high consistence. The result shows that tea tree response to pathogen infection is a complicated process. A number of genes were induced or suppressed. Disease-resistant transcription factors were highly activated and up-regulated. This study provided a theoretical basis for identifying tea resistance genes and potential molecular mechanism.

Determination of Glyphosate and Aminomethyl Phosphonic Acid Residue in Green Tea by Gas Chromatography-Mass Spectrometry

YANG Yaqin, FENG Shuhui, HU Yongjian, LI Yuanyuan, WANG Huifeng, LIU Jinxi, ZHONG Hongjian

Journal of Tea Science. 2020, 40(1): 125-132. doi:10.13305/j.cnki.jts.20191202.003

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An efficient method for the determination of glyphosate and its metabolite aminomethyl phosphonic acid (AMPA) in green tea was developed based on gas chromatography-mass spectrometry (GC-MS). Green tea samples were extracted with water, preliminary purified by dichloromethane and followed by purification with PCX and HLB combined solid phase extraction columns, derived with trifluoroacetic anhydride (TFAA) and 2,2,3,3,4,4,4-heptafluoro-1-butanol (HFB), then determined by GC-MS. For glyphosate, the limit of quantification (LOQ) was 0.05 mg·kg^-1, showing good linearity with coefficient R²=0.999 3 in the concentration range from 2-100 ng·mL^-1. For AMPA, itsLOQ was 0.02 mg·kg^-1, showing good linearity with coefficient R²=0.999 2 in the concentration range from 1-100 ng·mL^-1. At the spiked levels of 0.25 mg·kg^-1 and 0.50 mg·kg^-1, the average recoveries of glyphosate in green tea were 90.8% and 93.2%, with relative standard deviation (RSD) of 4.93% and 6.74%. At the spiked levels of 0.10 mg·kg^-1 and 0.20 mg·kg^-1, the average recoveries of AMPA in green tea were 85.8% and 95.4%, with RSD of 10.5% and 5.16%. The proper impurity purification, small impurity interference and high recovery rate made this method suitable for residue detection of glyphosate and its metabolite APMA in green tea.

Quantitative Analysis on Brand Premium Factors of Tea Enterprises Based on AHP and PSO

SHEN Xuezheng, JIN Yujie, SU Zhucheng

Journal of Tea Science. 2020, 40(1): 133-142. doi:10.13305/j.cnki.jts.20200117.005

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Brand premium is of great significance in the marketing operation of tea companies, and it is affected by many factors. Through 150 questionnaires and expert reviews, three first level factors, namely product quality, brand uniqueness and brand strength, and nine second level factors, including origin ecology, processing technology, production scale and brand personalized image were established. Meanwhile, the Analytic Hierarchy Process (AHP) was used to quantitatively analyze each factor. The Particle Swarm Optimization (PSO) was applied to improve the accuracy of results. A quantitative model of tea brand premium was built. It was found that brand awareness and origin ecology were the most important factors affecting brand premium. Then, according to different brand types, four brands of tea enterprises were selected to conduct empirical research, analyze their brand premium capabilities, and make recommendations.

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