Journal of Tea Science

Cloning and Expression Analysis of △12-fatty Acid Desaturase in Tea Plants

CHEN Dan, YU Ying, YUE Chuan, WANG Pengjie, CHEN Jing, CHEN Guixin, YE Naixing

Journal of Tea Science. 2017, 37(6): 541-550.

Abstract ( 309 )

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Based on the transcriptome database of tea plants, the cDNAs ( CsFAD2 and CsFAD6) including full ORFs of the key rate-limiting enzyme-FAD (△12-fatty acid desaturase) in the unsaturated fatty acid synthesis pathway were cloned from the buds and leaves of tea cultivar Tieguanyin by RT-PCR .The full length of cDNA of CsFAD2 was 1 184 bp, which contained a 1β149βbp ORF encoding 382 amino acids with 97% homologous to the CoFAD2. Subcellular localization prediction showed that CsFAD2 was localized to the endoplasmic reticulum. The full length of cDNA of CsFAD6 was 1β425βbp and contained a 1β311βbp ORF encoding 436 amino acids, which was 81% homologous to VvFAD6. It was predicted to be located on the chloroplast. Quantitative PCR analysis showed that the expression of CsFAD2 and CsFAD6 were induced by cold stress (4℃), which had the highest expression levels at 48βh. Similarly, CsFAD2 and CsFAD6 were induced by 100βg·L^-1 PEG treatment for 12βh. The expression of CsFAD2 was significantly up-regulated under the treatment of ABA(100βμmol·L^-1) for 6-24βh. While the expression of CsFAD6 was not affected by ABA during this period and dramatically down-regulated at 72βh. The expression of CsFAD2 was repressed under NaCl (250βmmol·L^-1) treatment. Inversely;|the expression of CsFAD6 was significantly induced by NaCl treatment from 24 to 72βh.

Cloning and Expression Analysis of miR156a-targeted Genes SPL6 and SPL9 in Camellia sinensis

LIU Yaqin, TIAN Kunhong, SUN Qilu, PAN Cheng, LI Yeyun, JIANG Jiayue, JIANG Changjun

Journal of Tea Science. 2017, 37(6): 551-564.

Abstract ( 337 )

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MicroRNA156a-SQUAMOSA promoter binding-like protein（miR156a-SPLs）play important roles in growth process, formation of leaves and response to stress in tea plant (Camellia sinensis). Based on transcriptome data, two cDNA CsSPL6 (2β318βbp) and CsSPL9 (1β954βbp) were cloned from tea plant. The nucleotide and amino acid sequences, biological information, phylogenetic tree and molecular expression models of the CsSPL6 and CsSPL9 were analyzed. Sequence analysis showed that the open reading frames of CsSPL6 and CsSPL9 are 1β668βbp and 1β116βbp, encoding 555 and 371 amino acids respectively. Both genes contain typical SBP domain and the recognition sites of microRNA156a. Quantitative real-time PCR analysis showed that the expression profile of Csn-miR156a was the highest in the buds, and the lowest in the 7th leaves, which was negatively correlated with CsSPL6 and CsSPL9. For different varieties(strains), the expression of Csn-miR156a was consistent with the growth of new shoots and the value of Photosynthesis index, but negatively correlated with the expression of CsSPL6 and CsSPL9, indicating. Csn-miR156a,CsSPL6 and CsSPL9 were involved in the growth of tea plant and can be used as markers to assess growth abilities among varieties. In addition, the expression of CsSPL9 was negatively correlatedwith Csn-miR156a under heat stress in four varieties (strains), which might be associated with the enhanced heat tolerance ability. The results showed that the expression of CsSPLs were negatively regulated by Csn-miR156a,which played an important role in the growth, development of tea plant and the stress resistance, providing a theoretical basis for understanding the growth and resistance mechanism of tea plant.

Cloning and Expression Analysis of CsBON3 from Copine Gene Family in Tea Plant (Camellia sinensis)

YAO Lina, HAO Xinyuan, WANG Lu, LI Nana, ZENG Jianming, YANG Yajun, WANG Xinchao

Journal of Tea Science. 2017, 37(6): 565-574.

Abstract ( 348 )

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Copine protein, a kind of Ca²⁺-dependent protein or phospholipid-binding protein containing two C2 (N-terminal) and one vWA (C-terminal) conserved domain, plays an important role in intracellular signal transduction. Based on the sequence identity analysis, an EST sequence showing high homology to Copine family genes was selected from our previous tea plant transcriptome database. The sequence was verified to contain 1β746βbp complete ORF, encoding 581 amino acids. Homologous alignment showed that the gene had the highest similarity with Arabidopsis AtBON3 (65%) and was named as CsBON3 (GenBank accession number: KY435900). Bioinformatic analysis showed that the molecular weight of CsBON3 protein was 63.66βkD and the theoretical isoelectric point was 5.48. It possesses the conserved domain unique to Copine family protein, which is a hydrophilic protein, no signal peptide site, non-secretory protein and no transmembrane domain. Expression analysis showed that CsBON3 had the highest expression levels in tea flowers and roots, and the lowest in stems and mature leaves. It was significantly up-regulated in tea plants treated with low temperature (4℃) for one day, and its expression was suppressed at dormant stage and up-regulated at growth stage. It was also up-regulated rapidly in the leaves with inoculation of Colletotrichum fructicola. The results indicated that the CsBON3 plays important roles in tea plant growth in response to low temperature and disease resistance.

Cloning of a MADS-box Transcription Factor Gene from Camellia sinensis and its Response to Abiotic Stresses

SHEN Wei, TENG Ruimin, LI Hui, LIU Zhiwei, WANG Yongxin, WANG Wenli, ZHUANG Jing

Journal of Tea Science. 2017, 37(6): 575-585.

Abstract ( 317 )

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In this study, based on the transcriptome database of tea plant, the CsMADS1 gene was cloned from cDNA of ‘Longjing43’ by RT-PCR method. The length of open reading frame of CsMADS1 gene was 657βbp, encoding 218 amino acids, which was a typical transcription factor of MADS-box family. Multiple alignments of CsMADS1 with related species showed that the identity of them was 65.65%, with a highly conserved MADS domain and a semi-conserved K domain. The physicochemical properties, hydrophilicity and hydrophobicity, subcellular localization, disordered feature, secondary and tertiary structure were also analyzed. CsMADS1 transcription factor is a hydrophilic protein, may be located in nucleus. The disordered feature of CsMADS1 protein was obvious, which was mainly composed of alpha helix structure, and had similar tertiary structure with MEF2 of human. Quantitative real-time PCR was used to analyze the expression profiles of the CsMADS1 gene under abiotic stress treatments of tea cultivar ‘Longjing43’. The results showed that the CsMADS1 gene responded to high and low temperatures, drought and high salinity. Different expression patterns of CsMADS1 gene were observed under those abiotic stress treatments.

Anti-obesity Efficacy of Shoumei White Tea on High-fat Diet Mice and Its Safety Evaluation

ZHENG Li, HOU Caiyun, REN Fazheng

Journal of Tea Science. 2017, 37(6): 586-596.

Abstract ( 350 )

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White tea extracts (WTEs) had been prepared from Shoumei white tea stored for 1, 3 and 7 years, respectively. Male mice (C57BL/6J) were fed with high-fat diet and supplemented with WTE to establish the anti-obesity efficacy model. Body weight gain, food intake, the pathobiology changes of the liver and adipose tissues, serum lipids profiles and differential expression profiles of lipid metabolism-related genes of liver in mice were investigated. The results showed that WTEs inhibited adipocyte differentiation and lipid excessive accumulation in adipose tissues, inhibited fatty acid synthesis and lipid excessive accumulation in liver by ameliorating the expression levels of lipid metabolism-related genes via down-regulation of the mRNA expression level of SREBP-1c, FAS and ACC1, thus depressing the increase of body weight and adipose tissues induced by high-fat diet. The preventing obesity efficacy of Shoumei white tea in high dosage was reduced with the increase of the storage years..

Simultaneous Determination of Perchlorate and Chlorate in Tea by Ultra-peryethylene Liquid Chromatography-tandem Mass Spectrometry

SONG Zhenggui, ZHANG Shufen, ZHOU Ziyan, ZHANG Aizhi, SHEN Jian, FU Xiao, LI Hesheng, XING Jiali, MAN Zhengyin, ZHAO Chunling, CAO Lili

Journal of Tea Science. 2017, 37(6): 597-604.

Abstract ( 329 )

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A method using ultra high performance liquid chromatography-tandem mass spectrometry was developed for the determination of perchlorate and chlorate in tea. Tea extracts were purified by ProElut C₁₈, separated by Click Xion column. The mobile phases were 5βmmol·L^-1 ammonium formate/water (A) and methanol: 5βmmol·L^-1 ammonium formate/water =9︰1 (B). The pH was adjusted to 3.2 by formic acid and the samples were then analyzed by UPLC-MS/MS with multiple reaction monitoring mode (MRM). The method was applied to detect perchlorate and chlorate in tea. The average recovery and relative standard deviations were in the range of 60.5%-85.8% and 4.9%-7.7%, respectively. The detecting limits of perchlorate and chlorate were 3βμg·kg^-1 and 5βμg·kg^-1, respectively. The method is simple, quick and high sensitive, which is suitable for the determination of perchlorate and chlorate in tea.

Study on the Chemical Constituents of Pu-erh Teas from Different Areas by UHPLC-Q-TOF/MS

YANG Chen, DAI Weidong, LYU Meiling, LI Pengliang, LIU Xu, TIAN Jun, WAN Yunlong, LI Ji, LIN Zhi

Journal of Tea Science. 2017, 37(6): 605-615.

Abstract ( 532 )

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In order to fully understand the influences of the production area on pu-erh raw tea quality and chemical composition, 12 pu-erh raw tea samples were selected from different mountains (villages) in Lincang, Pu'er, and Xishuangbanna. The nonvolatile tea metabolite phenotypes of the pu-erh raw teas were analyzed by ultra-high performance liquid chromatography quadrupole time-of-flight mass spectrometry-four (UHPLC-Q-TOF/MS). The results showed that the pu-erh raw teas from different areas had distinct chemical compositions. Pu-erh raw teas from Xishuangbanna (including Mengla, Menghai, and Jinghong), Pu'er, and Lincang could be seperated by principal component analysis. Pu-erh raw teas could also be seperated according to producing areas (southeast, southwest, and northwest of pu-erh tea area). Seventy nine main components in pu-erh raw teas were identified, and the relationships between the components and the taste qualities of the 12 pu-erh raw teas were also analyzed. This study showed that the metabolite phenotypes analyzed by UHPLC-Q-TOF/MS could be used for the discrimination of pu-erh raw teas according to producing areas.

Effects of Superfine Grinding on the Physicochemical Properties of Congou Black Tea

CHU Feiyang, CHEN Heli, SUN Dian, HE Huafeng, YE Yang, TONG Huarong

Journal of Tea Science. 2017, 37(6): 616-622.

Abstract ( 365 )

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In the study, the black tea sample was crushed into superfine grinding black tea powder with average particle size of 26.116, 9.612, 4.338, 3.742, 3.328βμm. The effects of superfine grinding on the physicochemical properties of congou black tea were investigated by means of determination of its materialized properties. The results showed that the particle size of superfine tea powder gradually decreased, the bulk density, expansion force and water holding capacity decreased, and the angle of repose and sliding angle gradually increased after superfine grinding, indicating that the fluidity was worse. The L value and b value of chroma gradually increase, which indicates that the superfine grinding can improve the color of the congou black tea. Embedded component test results showed that as the superfine tea powder particle size is reduced, water content of extract, amino acid content and soluble sugar content increased significantly, but the tea polyphenol, tea pigment and the content of crude fiber gradually reduce, It is presumed that the superfine grinding destroyed the cell wall structure of cellulose as the main body, and the crude fiber was damaged, thereby the contained ingredients were more easily exposed and the polyphenols were unstable and easily oxidized. Thus tea pigment content had a downward trend.

Studies on the Preparation and Stability of Tea Polyphenol Emulgel Formulation

SONG Juan, WANG Dong, YU Xudong

Journal of Tea Science. 2017, 37(6): 623-630.

Abstract ( 360 )

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The appearance properties, the particle size and stability of emulsion, the viscosity and content of emulgel were taken as evaluation indexes to optimize the best formula and preparation technology of tea-polyphenol emulgel and investigate its stability. The results indicated that the prepared emulgel was good in appearance, non-greasy, easy to spread and high stability. Moreover, the optimized preparation technology for emulgel was simple and the prepared tea polyphenol emulgel is a superior new preparation for topical external application.

Effect of Different Drying Technologies during Raw Tea Processing on the Quality of Green Tea Beverage

YUAN Haibo, HUA Jinjie, DENG Yuliang, LI Jia, DONG Chunwang, YANG Yanqin, WANG Jinjin, HUANG Yinquan, YIN Junfeng, JIANG Yongwen

Journal of Tea Science. 2017, 37(6): 631-637.

Abstract ( 480 )

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To explore the appropriate drying technology during raw tea processing, leaching behaviors and quality stabilities of tea samples prepared from raw materials dried by different methods or degree of drying were compared. The results revealed that (1) Raw tea dried by frying showed the highest contents of tea polyphenols when processed under low temperature, whereas those dried by baking-frying, or baking-frying-backing showed the highest contents of tea polyphenols when processed under high temperature. (2) In respect of color stability, tea soup prepared from baking-dried tea showed significantly elevated L value after sterilization or during storage, with the average L value increased by 2.035-3.905. Meanwhile tea soup prepared from baking-dried tea also showed higher anti-discoloration, with a/b value during low-temperature storage increased by 19.5%. (3) In respect to sensory flavor, most tea soups showed yellow-green color and ripe aroma while tea samples processed by baking-drying remained green, bright and fresh without any ripe taste. Besides, tea samples processed by baking-drying also showed superior flavor stability during storage irrespective of temperatures. In conclusion, baking approach during raw tea process was proposed with the optimized drying content between 5%-6%.

A New Insect Pest Crenidorsum turpiniae (Takahashi, 1932) in Tea Plant (Camellia sinensis) and Its Biology

MENG Zehong, WANG Jirui, ZHOU Xiaogui, LI Shuai, YANG Wen, WANG Hao, ZHOU Yufeng

Journal of Tea Science. 2017, 37(6): 638-644.

Abstract ( 384 )

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A whitefly species infesting tea plants, which took place frequently in Meitan County of Guizhou Province was firstly identified and initially observed. The specimens of puparia were observed by microscope and scanning electric microscope and they were identified as Crenidorsum turpiniae (Takahashi, 1932). It was firstly reported in tea plant. The morphological features and infestation characteristics were also described and illustrated.

Miang (Thai Fermented Tea) and Its Cuisine

BOUPHUN Tunyaluk, XU Wei, JIANG Yihe, ZHU Qi

Journal of Tea Science. 2017, 37(6): 645-652.

Abstract ( 1550 )

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Miang is an edible pickled tea or traditional tea leaf fermented product in household by local people in northern Thailand, which is made from the fresh tea leaves. This article went through the history of Miang, discussed its standard and method of plucking, stated the similar teas made in other Asian countries, such as China, Japan, Laos and Myanmar. Miang has long history and is very important for the northern Thai local economy. It also discussed its chemical constituents and the microorganisms responsible for the natural fermentation process as well as its benefits derived by humans. Challenges faced in the conducting its manufacturing industry and promoting it for better development in the future were also introduced.

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