Journal of Tea Science

Effects of the Combined Application of Biochar and Nitrogen on Growth and Nitrogen Use Efficiency of Tea Plants

WANG Feng, WU Zhidan, CHEN Yuzhen, JIANG Fuying, ZHU Liugang, ZHANG Wenjin, WENG Boqi, YOU Zhiming

Journal of Tea Science. 2018, 38(4): 331-341. doi:10.13305/j.cnki.jts.2018.04.001

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A pool experiment was conducted to study the effects of combined application of biochar and nitrogen fertilizer on tea plant growth and ¹⁵N transformation (Tree uptake, ammonia volatilization, N₂O emission and soil residual) using ¹⁵N trace technique. The results showed that the N application treatments improved the tea plant growth, and the tea yields of nitrogen application treatments increased from 68.06% to 112.63% as compared B₀N₀. Biochar effect on tea yields varied with N applications. Under N₀ and N₁ conditions, the tea yields of B₁N₀ and B₁N₁ treatments increased by 8.82% and 8.75%. But under N₂ conditions, the tea yield of B₁N₂ treatment was slightly reduced, although not significant. Compared with the B₀N₀, N application significantly increased the amounts of ammonia volatilization and N₂O emission. Under N₁ conditions, the B₁N₁ treatment reduced the ammonia volatilization and N₂O emission by 5.87% and 4.99% respectively. Under N₂ conditions, the B₁N₂ treatment reduced the ammonia volatilization and N₂O emission by 9.97% and 11.41% respectively. Compared with N treatment, the biochar mixed with N treatments increased the nitrogen concentration, ¹⁵N and the contribution of N derived from fertilizer (Ndff) in different parts of tea plants, thereby facilitated nitrogen uptake. The ¹⁵N use efficiency and ¹⁵N residue rate of biochar mixed with N treatments were higher than those of the N treatments with the percentages increasing by 0.46 to 3.93 percent and 4.09 to 14.37 percent respectively. The ¹⁵N loss rates of biochar mixed with N treatments were lower than those under N treatments, which were 4.54% to 18.30% differences, especially for B₁N₁treatment. In general, application of biochar in tea garden soil could promote N uptake in tea plants, increase the N fixed by soil and decrease N gaseous loss, thus improve the N use efficiency. Taken together, the results supported the theory of “less N fertilizer and increase yield” under B₁N₁ treatment, and this approach could be applied in tea production.

Organic Carbon and Total Nitrogen Distribution in Aggregates from Yellow-red Soils Under Tea Plantations with Different Fertilizer Managements

WANG Limin, HUANG Dongfeng, LI Qinghua, HE Chunmei, ZHANG Hui, LIU Cailing, LI Fangliang, HUANG Yibin

Journal of Tea Science. 2018, 38(4): 342-352. doi:10.13305/j.cnki.jts.2018.04.002

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Application of livestock manures and plant residues is a feasible practice to largely mitigate soil-degradative trends by increasing amounts of organic matter. However, long-term effects of fertilization regimes on the distribution of organic carbon (TOC) and total nitrogen (TN) in aggregate size fractions were poorly documented in tea (Camellia sinensis (L.) Kuntze) plantations in subtropical areas. A 10-year study was thus conducted to elucidate the impacts of fertilization patterns on the stability and allocation of TOC, TN and C/N ratio within water stable aggregate (WSA) fractions in soils under non-fertilization control (CK), chemical fertilizers only (CF), and integrated use of chicken manure and legume straw with chemical fertilizers (IF), respectively. Soil (0-20 cm) samples were collected and separated into ≥2, <2-1, <1-0.5, <0.5-0.25 and <0.25-0.106βmm aggregate classes by wet sieving, and finally used for TOC and TN analysis. The results showed that the IF treatment significantly (P<0.05) increased TOC and TN storage in all aggregate fractions relative to that of CK. Irrespective of the fertilization patterns, tea soils in subtropical China stored higher amounts of TOC and TN in the ≥2βmm water-stable fraction. In terms of the TOC and TN storage, the order of different aggregate fractions in IF- and CF-treated soils was (≥2β mm ) > (<0.5-0.25βmm) > (<1-0.5βmm) > (<2-1βmm) > (<0.25-0.106βmm), whereas that of CK-treated soils was (≥2βmm) > (<1-0.5βmm) > (<0.5-0.25βmm) > (<2-1βmm) > (<0.25-0.106βmm). Furthermore, the percentage of WSA ≥2βmm ranged from 54.95% under CK to 66.97% under IF treatment. Furthermore, TOC and TN adhered to WSA ≥2βmm, which accounted for 33.31%~45.45% of TOC and 33.76%~46.60% of TN in bulk soils under all treatments. Meanwhile, the IF treatment significantly (P<0.05) increased mean weight diameter (MWD) of aggregates as compared to that in CK- and CF- treated soils. However, a lower C:N ratio was observed in different aggregate fractions in IF-treated soils compared with that of CK. Additionally, WSA ≥2βmm was positively and significantly correlated with TN and TOC contents. In conclusion, the IF treatment improved aggregate stability, increased C and N storage in bulk soil and aggregates, and thus enhanced soil quality in tea farms in subtropical China.

Effects of Exogenous Salicylic Acid on Photosynthesis and Antioxidant Enzymes of Tea Plants under High Temperature

WEI Jipeng, LI Xin, WANG Zhaoyang, LI Yang, ZHANG Lan, SHEN Chen, YAN Peng, ZHANG Liping, HAN Wenyan

Journal of Tea Science. 2018, 38(4): 353-362. doi:10.13305/j.cnki.jts.2018.04.003

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In recent years, heat stress was more frequently occurred in tea gardens. However, few studies were focused on the approaches towards the improvement of heat tolerance in tea plants. In this study, Longjing 43 was used as experimental material to investigate the effects of various concentrations of MeSA on the net photosynthetic rate (Pn), the maximum carboxylation rate of Rubisco (V_c_,max), the maximum RuBP regeneration rate (J_max), electrolyte leakage (EL), MDA content and antioxidant enzyme activities in tea leaves under heat stress. Results showed that 1βmmol·L^-1 MeSA could significantly increase Pn, V_c,_max and J_max in tea leaves under heat stress. EL and MDA content in tea leaves increased significantly after heat stress, while the application of MeSA attenuated heat-induced increases in EL and MDA. In addition, the application of 1βmmol·L^-1 MeSA stimulated the activities of APX and CAT, leading to an efficient scavenging of hydrogen peroxide (H₂O₂) in tea leaves. In summary, we revealed that the applications of MeSA could improve photosynthetic capacity, strengthen the antioxidant system, reduce reactive oxygen species accumulation and lipid peroxidation in tea leaves under heat stress, and thus improve the tolerance of tea plants under heat stress.

Antioxidant Enzyme Activity of Tea Plant (Camellia sinensis) in Response to Low Temperature Stress

LIN Zhenghe, ZHONG Qiusheng, YOU Xiaomei, CHEN Zhihui, CHEN Changsong, SHAN Ruiyang, RUAN Qichun

Journal of Tea Science. 2018, 38(4): 363-371. doi:10.13305/j.cnki.jts.2018.04.004

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New clonal tea varieties 0306D, 0306F, 0306I from F1 population of Baijiguan and Huangdan were employed as test materials. Antioxidant enzyme activities of tea plants under low temperature were measured. The results showed that leaves under low temperature had higher conductivity, malondialdehyde (MDA) and hydrogen peroxide (H₂O₂) contents with the increasing ratios higher in 0306D and 0306F than 0306I and Huangdan. The increasing ratios of peroxidase (POD), superoxide dismutase (SOD), ascorbate peroxidase (APX) and glutathione reductase (GR) activities of the Huangdan and 0306I tea varieties were higher than 0306D and 0306F. While monodehydroascorbate reductase (MDAR), dehydroascorbate reductase (DHAR), ascorbate (ASC), reduced glutathione (GSH) and catalase (CAT) activities of 0306D and 0306F were less decreased than 0306I and Huangdan. The results indicated that higher activities of defense enzymes and higher antioxidant content in 0306I and Huangdan under stress were associated with chilling tolerance. Further regression analysis showed that the H₂O₂ content was negatively correlated with APX and CAT, and conductivity was negatively correlated with ASC content. The above results indicated that the tolerance to low temperature in 0306I and Huangdan was stronger than 0306F and 0306D.

Driving Factors and Their Acting Path of Farmers' Ecological Construction Behavior in Tea Garden ——Analyzed on Sample Data of Anxi County by SEM

GAO Shuilian, LEI Zhengyan, HU Shanshan, CHEN Qianjie, YANG Jiangfan

Journal of Tea Science. 2018, 38(4): 372-384. doi:10.13305/j.cnki.jts.2018.04.005

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To explore the driving factors and their acting path of famers’ ecological construction behavior in tea gardens, 310 effective survey data from Anxi County were obtained and a structural equation model (SEM) was established. The results showed that the policy factors, ecology consciousness and development expectation could promote tea farmers’ ecological construction behavior to effectively improve the ecological conditions of tea gardens. The influence coefficients of three key driving factors were 0.43, 0.36, 0.21, and the tea farmers’ ecological construction behavior could effectively improve the ecological situation of tea gardens, with the explained degree of about 0.87. Therefore, government should improve the tea garden ecological construction policies, enhance the ecological consciousness and ability of tea farmers, guide farmers to establish the correct concept of tea industry development and construct the guidance mechanism of ecological tea consumption market to drive the farmers' ecological construction behavior in tea garden.

The Identification and Analysis of Polyphenol Oxidase Gene Family in Tea Plant (Camellia sinensis)

ZENG Zeyuan, LUO Yong, ZHENG Chuchu, LI Juan, LI Qin, LIN Haiyan, WANG Kunbo

Journal of Tea Science. 2018, 38(4): 385-395. doi:10.13305/j.cnki.jts.2018.04.006

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：Polyphenol oxidase (PPO) is a copper-containing metal enzyme, which plays an important role in tea processing. Five PPO genes were found in the tea genome database: CsPPO1, CsPPO2, CsPPO3, CsPPO4 (GenBank Access. No. GQ129142.1) and CsPPO5. The nucleic acid and amino acid sequences of five PPO genes were analyzed, and the physicochemical properties and structural functions of the PPO genes encoding protein were also predicted by bioinformatics method. The results showed that the full length of the five PPO genes coding sequences (CDS) was between 597βbp and 1β839βbp which could encode 198-612 amino acids. Phylogenetic tree analysis showed that CsPPO1, CsPPO3, CsPPO4 and CsPPO5 had close genetic relationships. The encoded proteins belonged to hydrophilic and unstable lipid binding proteins and had no signal peptides. Irregular curl was the main structural element of protein in which α-helix and β-strand were dispersed throughout the polypeptide chain & CsPPO1, CsPPO2, CsPPO3 might contain one or more transmembrane spirals. The five PPO proteins contained PPO1_DWL (pfam12142) conserved structural domains. It was predicted that the proteins encoded by CsPPO1, CsPPO3, CsPPO4 and CsPPO5 might contain tyrosinase functional domains. Real-time reverse transcription (qRT-PCR) analysis showed that the PPO genes showed different expression patterns among different tea cultivars.

Effect of Two Prokaryotic Expressed Vectors on the Activity of PPO from Camellia sinensis

GAN Yudi, SUN Kang, LI Huijuan, DU Zhongying, ZHAO Zhen, PANG Xing, LI Xinghui, CHEN Xuan

Journal of Tea Science. 2018, 38(4): 396-405. doi:10.13305/j.cnki.jts.2018.04.007

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To get stably soluble tea polyphenol oxidase with high activity, two vectors were selected to express CsPPO. A tea polyphenol oxidase gene was cloned from tea leaves. After trimming two peptide fragments with 43 and 70 amino acids, the sequences were connected into pET32a and pMAL-c5X vectors with BamH I/xho I and Sal I/ BamH I and named as pET32a-CsPPO₄₃, pET32a-CsPPO₇₀, pMALc5X-CsPPO₄₃ and pMALc5X-CsPPO₇₀ respectively. After expressed of recombinant plasmids in the E.coil Transetta (DE3) strain, SDS-PAGE results showed that the protein expressed by pMAL-c5X was easier extracted than that by pET32a. The proteins expressed by pMAL-c5X were found in both the supernatant and pellet, while those by pET32a were only found in the pellet. The activity of CsPPO was detected by 1,2-benzenediol, ECG and EC as substrates in 410 nm with micro-plate reader. The ‘S’ shaped curves graphed with three substrates oxidized by the pMALc5X-CsPPO indicated that all reaction speeds were not constant. The specific activity of pMALc5X-CsPPO₄₃ was higher than that of pMALc5X-CsPPO₇₀, with the highest activity reaching 1.45×10⁶βU·mg^-1 when reacted to EC. Therefore, pMALc5X-CsPPO₄₃ could be used to synthesize industrial PPO, but its role in catechin mechanism still needs further research.

Discrimination of Fermenting Degree of Congou Black Tea Based on Monoterpenoid

HE Huafeng, GUI Anhui, YE Yang

Journal of Tea Science. 2018, 38(4): 406-409. doi:10.13305/j.cnki.jts.2018.04.008

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As the basic constituents, monoterpenoids are important indicators for the tea aromatic quality. The time course changes of aromatic components were tracking analyzed throughout the fermentation process. Based on organoleptic evaluation, degree of fermentation was classified qualitatively. Using gas chromatography-mass spectrometry (GC-MS), compositions of aroma were screened during fermentation, and 21 monoterpenoids were discriminated by taking chemical structure into consideration. Employing partial least squares discrimination analysis (PLS-DA) to analyze, effective prejudgment of the fermenting degrees was obtained. Five compounds of the monoterpenoids were calculated as the crucial compositions in connection with the prognosis. Result of the research would be the theoretic guarantee for the feedback control of fermentation process of Congou black tea.

Design and Experiment of Tea Far Infrared Dryer Based on DMC-PID Cascade Control

LI Bing, SUN Changying, LI Weining, SONG Yangyang

Journal of Tea Science. 2018, 38(4): 410-415. doi:10.13305/j.cnki.jts.2018.04.009

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According to the characteristics of delay, inertia and nonlinearity in the baking of tea materials, the low accuracy of temperature control, large overshoot and poor robustness of traditional PID control largely limited relative application. A tea drying machine was designed based on DMC-PID control in this paper. The drying machine adopted multi-layer tunnel with the infrared radiation plate heating element above the conveyor belt. The DMC-PID cascade temperature control system was also used. The first DMC algorithm improved the robustness and capability of the dynamic temperature control system and the second PID algorithm improved the anti-jamming performance simultaneously. The green tea baking experiment on the prototype was carried out. The results showed that when the PID temperature control system was adopted, the overshoot of the dryer became 10.5%. When the DMC PID cascade temperature control was used,the overshoot was 5.9%. DMC-PID control could improve the temperature control precision and tea quality of the tea dryer.

Determination of Phthalimide Residue in Tea Using Gas Chromatography-tandem Mass

GAO Guanwei, CHEN Hongping, CHAI Yunfeng, JIN Lili, LIU Xin, LU Chengyin

Journal of Tea Science. 2018, 38(4): 416-424. doi:10.13305/j.cnki.jts.2018.04.010

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As folpet residues, phthalimide (PI) and folpet were responsible for a high risk of false positive detection of folpet in tea, which became a major factor restricting the export of Chinese tea. A modified QuEChERS method for the determination of PI residue in tea using gas chromatography-tandem mass was developed and validated. The target analyte was extracted using acetonitrile and the crude extracts were purified using multiwalled carbon nanotubes, octadecyl silica and strong cation exchanger. At the spiked levels of 10, 20, 50 and 100βμg∙kg^-1, the average recoveries were from 73% to 104%, and the relative standard deviations were below 20%. The limit of quantification of PI in tea was 10.0βμg∙kg^-1. The method was simple, reliable, accurate, sensitive and appropriate for the determination of PI residue in tea.

Determination the Calorific Value of Tea by Bomb Calorimeter

GAO Jinhong, CHEN Ying, LI Yali

Journal of Tea Science. 2018, 38(4): 425-429. doi:10.13305/j.cnki.jts.2018.04.011

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The calorific value is an important physical data to measure the quality of materials. It can provide a new idea and method for evaluating the quality of similar foods from the energy point of view. The calorific values of 12 kinds of tea samples were determined by bomb calorimeter in this study. The results showed that the calorific values of these tea foods ranged from 14.96βkJ·g^-1 to 16.16βkJ·g^-1. The determining method was simple, easy to operate. Understanding the calorific value of tea had a certain guiding and referencing significance for the development, utilization, storage and consumption of tea foods.

Alleviative Effects of Aged Liupao Tea on Lipid Metabolism and Liver Injury in Hyperlipidemic Mice

WU Wenliang, LIU Zhonghua, LIN Yong, HUANG Jian′an, ZUO Gaolong, TENG Cuiqin, LONG Zhirong, QIU Ruijin, CAO Zhonghuan

Journal of Tea Science. 2018, 38(4): 430-438. doi:10.13305/j.cnki.jts.2018.04.012

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To investigate the alleviative effects of aged Liupao tea on lipid metabolism and liver injury in hyperlipidemic mice, the changes of main chemical substances in Liupao tea of different storage ages were analyzed. The hyperlipidemia model mice were established by feeding a high fat and sugar diet, and Liupao tea groups were given daily oral administration of water extract of selected aged Liupao tea. The changes of related physiological and biochemical indexes and tissue morphology in mice were observed. The results showed that Liupao tea samples stored for 15 years were selected as the representative of the aged Liupao tea, which was used in subsequent experiments. Compared with the hyperlipidemia model group, aged Liupao tea especially medium or high dose groups could significantly decrease the contents of TC, TG, and LDL-C levels, increase the content of HDL-C, suppress the activities of ALT and AST in the serum, and moderate liver oxidative stress,. Pathological sections of liver and adipose tissue showed that aged Liupao tea could reduce the formation of lipid droplets in liver cells and inhibit the enlargement of fat cells in a dose dependent manner. In conclusion, aged Liupao tea of normal dose could effectively improve lipid metabolism disorder and liver injury in mice induced by high fat and sugar diet.

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