Journal of Tea Science

Research Progress on DNA Molecular Fingerprinting of Tea Plant (Camellia sinensis)

HUANG Danjuan, MA Jianqiang, CHEN Liang

Journal of Tea Science. 2015, 35(6): 513-519. doi:10.13305/j.cnki.jts.2015.06.001

Abstract ( 483 )

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Molecular fingerprintings reveal the differences between individuals at DNA level directly, which can identify tea cultivars effectively and protect the right of the variety plant. In this paper, the classification, concept and characteristics of fingerprinting were described. The research progress included 7 technologies for tea plant fingerprinting, as well as the application of fingerprinting in tea germplasm resources and breeding investigation were introduced. The problems existed in fingerprinting research of tea plant were discussed, and the construct of standard fingerpinting of tea cultivars was proposed.

Effects of Exogenous Calcium and Inhibitors of Calcium Signaling Transduction Pathway on Cold Resistance of Tea Plant

HUANG Yuting, QIAN Wenjun, WANG Bo, CAO Hongli, WANG Lu, HAO Xinyuan, WANG Xinchao, YANG Yajun

Journal of Tea Science. 2015, 35(6): 520-526. doi:10.13305/j.cnki.jts.2015.06.002

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The calcium signal transduction pathway plays a pivotal role in the response to the stress of low temperatures. In order to verify the relationship between calcium signal transduction pathway and cold resistance of tea plant, the changes of some cold resistance related index were measured under artificial low temperature stress in climatron accompanied with using exogenous CaCl₂, CaM inhibitor W-7 [N-(6-Aminohexyl)-5-chloro-1-naphthalene-sulfonamide] and calcium signal channel inhibitor LaCl₃, respectively. Calcium signaling transduction pathway inhibitors W-7 and LaCl₃ improved electrolyte leakage, enhanced contents of malondialdehyde (MDA), superoxide anion radical (O₂·^-) and proline, reduced the activities of superoxide dismutase (SOD) compared with control treatment, however, CaCl₂ treatment showed opposite effects. It showed that calcium signaling system play an important role in the response to low temperature stress in tea plant.

Morphological and Histological Observations on the Reproductive System in Male Adults of Ectropis obliqua Prout

ZHANG Jiaxia, SUN Qingyu, ZHAO Qiang, XIA Xianjiang, DING Yong

Journal of Tea Science. 2015, 35(6): 527-533. doi:10.13305/j.cnki.jts.2015.06.003

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By using both routine anatomy and paraffin-section microscopy, the morphology and histology of internal reproductive organs and external reproductive organs of the male adults in Ectropis obliqua Prout were described. Like other major organizational structure in Lepidoptera, the external organs including harpes, tegument, uncus and aedeagus, the internal organs consisted of a testis, a pair of seminal vesicle, a pair of accessory glands, and ductus ejaculatorius simplex. Eight radial testicutar tubule which were separated by thinner connective tissue were also observed in testis slices. And internal thinner connective tissue distributed a large number of spermatogonia, spermatocytes and mature sperm bundles. The results found seminal vesicles and vas deferens were filled with liquid and also showed a large quality of sperm bundles in the seminal vesicles. This is the first study to clarify the male reproductive system and sperm bundles filling state in the thinner connective tissue of this species and it provides more guiding significance for field forecast, phylogeny and species identification.

Cross-resistance Induced by Tea Polyphenols in Pseudomonas aeruginosa

LIU Xiaoxiang, SUN Aihua, DU Peng, CHEN Wenhu, ZHU Junli

Journal of Tea Science. 2015, 35(6): 534-542. doi:10.13305/j.cnki.jts.2015.06.004

Abstract ( 555 )

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Cross-resistance induced by sublethal concentration of tea polyphenols (TP) to Pseudomonas aeruginosa was investigated. One-hour exposure of Pseudomonas aeruginosa to 1βmg·mL^-1 TP significantly increased the tolerance to multiple environmental stresses, including oxidant (1βmmol·L^-1 H₂O₂), high temperature (47℃) and acid solutions [ phosphate buffer (pH4.0), phosphate buffer (pH4.0) containing 60βmmol·L^-1 citric acid, 60βmmol·L^-1 lactic acid and 80βmmol·L^-1 acetic acid]. In addition, the effects of TP on expression levels of a few stress-related genes were studied by quantitative real-time RT-PCR analysis. It was observed that the expression levels of a few oxidative stress-related genes katB, sodM, ohr, lexA and recN were up-regulated by TP, as well as some heat shock protein genes, including dnaK, groEL, htpG, grpE and groES. The expression of these genes might play important roles in the development of stress cross-resistance. Although TP was considered to be safe as food preservatives, the underlying hazards associated with cross-resistance induced by TP should be taken into account in food technology.

Effects of Exogenous 24-epibrassinolide on the Photosynthetic Characteristics of Tea Plants (Camellia sinensis)

LI Zhixin, LI Xin, FAN Lichao, HAN Wenyan

Journal of Tea Science. 2015, 35(6): 543-550. doi:10.13305/j.cnki.jts.2015.06.005

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24-epibrassinolide (EBR) is an important plant hormone, which plays an important role in a variety of adverse resistance in plants. The effects of exogenous EBR on photosynthesis, Rubisco and FBPase activity and the expression of related-gene from tea leaves (Camellia sinensis cv. ‘Longjing 43’, ‘Xiangguliaobaihao’, and ‘Qingmingzao’) under field condition were investigated. It was showed that after exogenous EBR (0.1βmg·L^-1) treatment: Net photosynthetic rate of these three cultivars increased by 49.06%, 45.49% and 92.34% respectively. Vc_max increased by 21.82%, 21.68 % and 21.68%, and J_max increased by 17.16%, 23.86% and 23.86%, respectively. The activity of Rubisco and FBPase increased significantly. Furthermore, the expression of Rubisco and FBPase related-gene increased significantly. These results suggested that application of exogenous EBR could enhance Vc_max, J_max and the activity of Rubisco and FBPase effectively, which ultimately promoted the photosynthetic carbon fixation, and the photosynthetic rate of tea plants.

Analysis on the Changes of Color of Withering Leaf in Different Environmental Humidity Conditions

YUAN Haibo, HUA Jinjie, DENG Yuliang, XU Yongquan, CHEN Gensheng, WANG Fang, JIANG Yongwen, YIN Junfeng

Journal of Tea Science. 2015, 35(6): 551-558. doi:10.13305/j.cnki.jts.2015.06.006

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Different withering humidity conditions are set with artificial climate box, the dynamic change of chromatic aberration during withering leaves and its inherent law were investigated. The results showed that, three chroma index L, a, b showed significant changes with the increase of withering time under the conditions of three different humidity treatments, and there were significant differences between different humidity treatments, but its difference was decreased gradually with the increase of the withering time. The law of change of L, a. b and overall color during tea withering process with the conditions of three different humidities were analysed in this investigation. The results showed that 70% humidity was regarded as the optimum humidity in the withering of tea. The results of this investigation provided the reference to the theoretical basis on the changes of physic-chemical properties in tea process.

Effects of LED Yellow Light on the Expression Levels of Aroma Related Genes and the Enzyme Activity in Withering Process of Congou Black Tea

XIANG Lihui, LIN Fuming, SUN Weijiang, WU Yang, CHEN Mingzhao, ZHANG Qinfang, ZHOU Lele, WENG Rui

Journal of Tea Science. 2015, 35(6): 559-566. doi:10.13305/j.cnki.jts.2015.06.007

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Zhenghedabaicha (Camellia sinensis) samples were divided into two groups of LED yellow light withering and no light withering as the control group both with Congou black tea processing. The withering leaves were collected to detect the expression level of β-glucosidase (CsBP), activities of β-glucosidase, aroma components of Congou black tea and the sensory evaluation were conducted to discuss the effects of LED yellow light on the quality of tea. Results showed that during LED yellow light withering,the expression levels of β- glucosidaseⅠ, Ⅱgenes were significantly improved, the expression level of β-glucosidaseⅡ gene was the highest at the 6th hour, which significantly higher than the control group. CsBP was up-regulated expressed in withering process. The rise of genetic expression is started from the Early LED withering stage, and the activity of enzyme increased in the later period of withering, which positively promoted the development of sweet and floral aroma of Congou black tea and finally the quality was improved and better than the control.

Correlation of Low Temperature with Soluble Sugar and Amino Acid Content in Fresh Tea Leaves

TIAN Ye, WANG Mengxin, WANG Jinhe, HAN Baoyu

Journal of Tea Science. 2015, 35(6): 567-573. doi:10.13305/j.cnki.jts.2015.06.008

Abstract ( 758 )

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In order to evaluate the substantial basis of tea plant resisting frost as well as the difference in frost resistance of different cultivars, the tea plantation growing with cultivar Wuniuzao and another plantation growing with Maolü were selected, in which the temperatute was recorded day by day. At the fifth, the fifteenth and the twenty fifth of each month during the experimental duration, the fresh tea leaves were plucked, in which the soluble sugar content was detected by colourimetry, and the proline content and seventeen free amino acid contents were determined by HPLC. The results showed that: ① Within the three durations the temperature dropped continually (from October 20 to December 27 in 2013), the low temperature lasted (from December 28 to February 20), and the temperature rose gradually, accordingly, the soluble sugar content continually raised, and keeped on increasing, then reduced insignificantly, furthermore, the soluble sugar content negatively correlated with temperature in the whole experimental stage. ② Accordingly, the proline content and the seventeen free amino acid content slowly raised, and obviously increased, then markedly promoted. It was considered that: ① The three types of substances were all related to the resisitance on cold injury, and the correlation of the soluble sugar content with low temperature was larger than those of other two types of substances with the temperature. ② The content of these three types of cold-resisting

Analysis of the Relationship between Soil Nutrients and Tea Main Quality Components of Ancient Tea Arboretum and Modern Tea Garden in Yunnan Province

YANG Guangrong, WANG Xiuqing, XIE Jin, LÜ Caiyou, LI Yongmei

Journal of Tea Science. 2015, 35(6): 574-582. doi:10.13305/j.cnki.jts.2015.06.009

Abstract ( 744 )

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Soil is the basic survival condition of the tea plant, so it was very closely related to the growth and development of tea plant and tea quality. The representative ancient tea arboretum and modern tea garden were selected in Jingmai, Bulang and Nannuo ancient tea mountain of Yunnan Province. The pH value, cation exchange capacity (CEC), soil organic matter (SOM), and nutrients content of nitrogen, phosphorus and potassium (NPK) in tea garden soils (0-20βcm) and the tea main quality component of sundried green teas which were manufactured by a bud and two leaves (water extraction, tea polyphenols, amino acid, caffeine and polyphenols monomer content) were determined from the soil and tea samples and those from the modern tea garden (or young tea tree/forest) were used as control. The results showed that the soil moisture content increased by 15.23%~46.82% in ancient tea garden under forests shade in comparing with that from modern tea garden. The soil pH value in the ancient tea garden was higher than that of modern tea garden (or young tea tree/forest) in Bulang mountains and Nannuo mountain. The soil CEC, SOM, total nitrogen (TN), total phosphorus (TP) and available phosphorus (Olsen-P) contents of ancient tea garden were significantly higher than those of modern tea garden (or young tea tree/forest). All though the contents of tea water extract, amino acid, tea polyphenol and polyphenols monomer substance in all tea gardens were not consistent, however, the ratio of polyphenol to amino acid of ancient tea plants planted under forests range was 6.75 to 12.28 significantly lower than those of the modern tea garden (from 9.98 to 15.25). The changes of five kinds of catechins content were in the order: Epicatechin gallate (ECG) > Epigallocatechin gallate (EGCG) > Epicatechin (EC) > Epigallocatechin (EGC) > Catechin (C), and the ECG and EGCG contents were not significant differed between ancient tea garden and modern tea garden (or young tea tree/forest). Moreover, tea gallic acid (GA) was detected only in the ancient tea plants. The cultivation pattern in ancient tea garden was beneficial to prevent the soil acidification, to reduce the illumination intensity, to improve the soil moisture content, and to promote the sustainable utilization soil fertility in tea garden, and it is advantageous to tea plant in forming high amino acid content and low ratio of polyphenol to amino acid.

The Research on Method of Identifying New Green Tea and Stale Green Tea Based on Characteristic Aromatic Components of Stale Tea Aroma

BAO Zhongzan, DONG Rongjian, DENG Zhaopu, XIE Bingtao

Journal of Tea Science. 2015, 35(6): 583-588. doi:10.13305/j.cnki.jts.2015.06.010

Abstract ( 633 )

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For the purpose of studying and confirming the characteristic aromatic components of stale tea to distinguish the new green tea from the stale Wenzhou green tea, the simultaneous distillation extraction—gas chromatography—mass spectrometry (SDE-GC/MS) was used to analyze seven kinds of tea samples from seven areas of Wenzhou. The results showed that (z)-2-penten-1-ol, 2,4-Heptadiene-1-al and 3,5-Octadien-2-one were not detected in the new tea, however, the three compounds could be detected and reveal a regular increasing tendency with the tea getting stale by going through normal temperature storage, refrigerated storage and frozen storage. Especially, the relation between the content of (z)-2-penten-1-ol and storage duration differed significantly under different storing condition. By employing research instruments such as standard curve, recovery rate, precision and detecting limit, we found that (z)-2-penten-1-ol could be an effective distinguishing indicator of new and stale tea, and 2,4-Heptadiene-1-al and 3,5-Octadien-2-one could be used as auxiliary identifying material, which laid the theoretic foundation of and provided data support to the identification of new and stale Wenzhou green tea.

Re-dissolution and Reclaim of Green Tea Sediment Based on the Treatment of Tannase

XU Yongquan, HU Xiongfei, CHEN Jianxin, CHEN Gensheng, WANG Fang, YIN Junfeng

Journal of Tea Science. 2015, 35(6): 589-595.

Abstract ( 481 )

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The formation of tea sediment not only influences the flavor and exterior quality of tea beverage or tea concentrate, but also causes a great loss of chemical components. Gallated catechins are the principal components participating in green tea sediment formation. This study discussed the effect of tannase treatment on the re-dissolution and reclaim of green tea sediment by hydrolyzing gallated catechins with tannase. The results showed that, the tannase treatment helped re-dissolve and reclaim of green tea sediment; the re-dissolution of sediment was enhanced with the increase of added enzyme amount and enzymatic time. 35℃, 150βmin and 2.0% of solids were obtained as the optimal parameters of tannase treatment by orthogonal test. Tannase treatment with optimal parameters could hydrolyze 98% gallated catechins and reduce 82% re-sediment. And tannase treatment also helped to improve the taste quality of the resolution of sediment and tea concentrate. This study will provide theory evidence for solving the problem of sediment in tea beverages.

Screening of microRNA Reference Genes for Real-time Fluorescence Quantitative PCR under Cold Stress in Camellia sinensis

XIE Xiaofang, TIAN Xianfeng, JIANG Changjun, LI Yeyun

Journal of Tea Science. 2015, 35(6): 596-604. doi:10.13305/j.cnki.jts.2015.06.013

Abstract ( 447 )

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MicroRNA (miRNA) plays an important role in post transcriptional regulation of gene expression. The research of miRNA quantitative expression level has become more and more popular. Choosing a suitable reference gene is a prerequisite of an accurate real-time fluorescence quantitative PCR(qRT-PCR) assay. The expressions of 9 candidate reference genes by qRT-PCR method in different tea (Camellia sinensis) samples were investigated and their stabilities were analyzed by BestKeeper and geNorm. The results showed that a novel tea miRNA (PC-3p-222) was the most stable reference gene in different tea plant tissues under different cold temperature. The average Ct value was 22-23 which showed moderate expression abundance. Therefore, it can be used as the appropriate reference gene for miRNA qRT-PCR under cold stress, which provides a good choice for the quantitative of miRNA expression.

Research on Preparation and Antitumor Activity of EGCG Naonparticles

HUANG Meirong, YING Hao, JIANG Yongwen, JIANG Heyuan, DU Qizhen

Journal of Tea Science. 2015, 35(6): 605-612. doi:10.13305/j.cnki.jts.2015.06.015

Abstract ( 441 )

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(-)-Epigallocatechin-3-gallate (EGCG) and ascorbic acid (Vc) were loaded in heat treated β-lactoglobulin (β-Lg) to prepare EGCG naonparticles. The effects of EGCG and Vc with different molar ratios on the particle size, Zeta potential, entrapment efficiency, loading capacity, the change of color were investigated. The antitumor activity and mechanism of EGCG naonparticles were obtained by MTT assay and cytokinesis block micronucleus test (CBMNT). The results showed that EGCG naonparticles had stronger inhibition on human melanoma cell (A-375) and esophageal carcinoma cell (TE-1), and significantly increased the number of micronuclei, necrosis and apoptosis of the A-375 and TE1-1 carcinoma cell.

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