茶树CsNCED2基因的克隆和表达分析

该研究以铁观音茶树品种叶片为材料,通过RT-PCR技术,克隆了茶树脱落酸(ABA)合成途径关键限速酶——9-顺式环氧类胡萝卜素裂解双加氧酶(9-cis-epoxycarotenoid dioxygenase,NCED)基因的全长cDNA序列。该基因cDNA全长1 931bp,包含1 821bp完整开放阅读框,共编码606个氨基酸残基。NCBI同源分析结果表明,与葡萄VvNCED2相似性最高(78%),命名为CsNCED2(NCBI登录号:MF765770)。氨基酸序列分析显示,其具有NCED家族的FLNO2258保守结构域,以及MIAHPKxDP和HDFAITE保守结构域序列;在保守区存在4个Fe2+活性组氨酸结合位点,N-端含有叶绿体转运肽。实时荧光定量PCR分析表明,CsNCED2基因在铁观音叶、茎和花中表达量较高;白茶萎凋和乌龙茶做青均可以诱导CsNCED2基因显著上调表达;除干旱胁迫抑制CsNCED2表达外,ABA和低温胁迫均能够诱导CsNCED2基因显著上调表达。表明CsNCED2基因在茶树ABA合成代谢以及胁迫响应中发挥重要作用。     

人工接种发酵茯砖茶降脂减肥作用研究

比较自然发酵与人工接种发酵茯砖茶对营养性肥胖大鼠的降脂减肥作用。结果表明与肥胖模型组比较,喂饲两类茯砖茶的大鼠体重、脂肪湿重、脂肪系数、甘油三酯（TG）、总胆固醇（TC）等指标均低于肥胖模型组,而高密度脂蛋白（HDL-C）则有明显提高。人工接种发酵茯砖茶与自然发酵茯砖茶均对营养性肥胖大鼠有降脂减肥作用,能显著改善大鼠的各项肥胖指标,人工接种发酵的茯砖茶在相关指标上均接近甚至优于自然发酵的茯砖茶。     

Effect of drinking green tea on age-associated accumulation of Maillard-type fluorescence and carbonyl groups in rat aortic and skin collagen.

Tea catechins and other flavonoids have been shown to potentially protect against chronic cardiovascular diseases such as coronary heart disease and atherosclerosis. In this study, 6-month-old female Sprague-Dawley rats were fed green tea extract (50 mg/100 ml in drinking water) up to the age of 22 months, and the age-associated changes in Maillard-type fluorescence and carbonyl groups in the aortic and skin collagen were compared with those occurring in the water-fed control animals. Collagen-linked Maillard-type fluorescence was found to increase in both the aortic and skin tissues as animals aged. The age-associated increase in the fluorescence in the aortic collagen was remarkably inhibited by the green tea extract treatment, while that occurring in the skin collagen was not significantly inhibited by the treatment. The collagen carbonyl content also increased in both the aortic and skin tissues as animals aged. In contrast with the case of Maillard-type fluorescence, however, the age-associated increase in the carbonyl content was not inhibited by the green tea extract treatment either in the aortic or skin collagen. These results suggest that the inhibition of AGE formation in collagen is an important mechanism for the protective effects of tea catechins against cardiovascular diseases.     

RNA-seq analysis of green tea polyphenols modulation of differently expressed genes in Enterococcus faecalis under low pH

Enterococcus faecasslis (E. faecalis) is a resident bacterium in the host. The increase in internal stress like low pH may affect the biological effects of E. faecalis. The prebiotic-like function of tea polyphenols can enhance the beneficial effects of its tolerance to environmental stress. In this study, RNA-sequence analysis was used to explore the protective effect of green tea polyphenols (GTP) on E. faecalis under low pH stress. A total of 28 genes were found to be responsive to GTP under low pH stress, including 16 up-regulated and 12 down-regulated. GTP intervention can partly relieve some undesired negative influences, such as the down-regulation of the base excision repair gene and amino acid transport and metabolism gene. The significantly changes were associated with selenocompound metabolism and aminoacyl-tRNA biosynthesis after the intervention of GTP. The present study provided new insights into the growth and continuous adaptation of E. faecalis under stress.     

cis- and trans-Linalool 3,7-Oxides and Methyl Salicylate Glycosides and (Z)-3-Hexenyl β-D-Glucopyranoside as Aroma Precursors from Tea Leaves for Oolong Tea

Two new alcoholic aroma precursors, cis- and trans-linalool 3,7-oxides 6-O-β-D-apiofuranosyl-β-D-glucopyranosides (1 and 2), as well as two already known compounds, (Z)-3-hexenyl β-D-glucopyranoside (3) and methyl salicylate 6-O-β-D-xylopyranosyl-β-D-glucopyranoside (β-primeveroside: 4), and another new monoterpendiol glycoside, 8-hydroxygeranyl β-primeveroside (5) have recently been isolated as aroma precursors in tea leaves (Camellia sinensis var. sinensis cv. Maoxie) ready for oolong tea processing.     

茶树中咖啡碱抑茵抗虫作用的研究

咖啡碱是茶树的主要生物碱和特征性物质之一,通过滤纸片法、生长速率法、饲喂称重法对咖啡碱抑菌、抗虫作用的研究,发现咖啡碱对供试细菌和真菌有一定的抑菌能力,对棉铃虫和菜粉蝶、家蚕幼虫具有毒害和抑制生长的影响。咖啡碱作为茶树的次生代谢产物具有抗病抗虫的的生物学功能。     

Molecular pathway for (-)-epigallocatechin-3-gallate-induced cell cycle arrest and apoptosis of human prostate carcinoma cells

Epigallocatechin-3-gallate (EGCG), the major polyphenolic constituent present in green tea, is a promising chemopreventive agent. We recently showed that green tea polyphenols exert remarkable preventive effects against prostate cancer in a mouse model and many of these effects are mediated by the ability of polyphenols to induce apoptosis in cancer cells [Proc. Natl. Acad. Sci. USA 98 (2001) 10350]. Earlier, we showed that EGCG causes a G0/G1 phase cell cycle arrest and apoptosis of both androgen-sensitive LNCaP and androgen-insensitive DU145 human prostate carcinoma cells, irrespective of p53 status [Toxicol. Appl. Pharmacol. 164 (2000) 82]. Here, we provide molecular understanding of this effect. We tested a hypothesis that EGCG-mediated cell cycle dysregulation and apoptosis is mediated via modulation of cyclin kinase inhibitor (cki)-cyclin-cyclin-dependent kinase (cdk) machinery. As shown by immunoblot analysis, EGCG treatment of LNCaP and DU145 cells resulted in significant dose- and time-dependent (i) upregulation of the protein expression of WAF1/p21, KIP1/p27, INK4a/p16, and INK4c/p18, (ii) down-modulation of the protein expression of cyclin D1, cyclin E, cdk2, cdk4, and cdk6, but not of cyclin D2, (iii) increase in the binding of cyclin D1 toward WAF1/p21 and KIP1/p27, and (iv) decrease in the binding of cyclin E toward cdk2. Taken together, our results suggest that EGCG causes an induction of G1 phase ckis, which inhibits the cyclin-cdk complexes operative in the G0/G1 phase of the cell cycle, thereby causing an arrest, which may be an irreversible process ultimately leading to apoptotic cell death. This is the first systematic study showing the involvement of each component of cdk inhibitor-cyclin-cdk machinery during cell cycle arrest and apoptosis of human prostate carcinoma cells by EGCG.     

色彩对茶园昆虫的引诱力

为评估色板用于监测和诱捕茶园害虫的可能性,于10月份茶园昆虫盛发期在绍兴和杭州分别选有机、无公害茶园,同时使用10种粘性色板大面积诱捕茶园昆虫和蜘蛛.每种色彩重复10次.结果表明:(1)在两片供试茶园捕获的目、科和种类基本相同.蜘蛛很少.24 h后,绍兴茶园中10种色板共捕获9目53科10200个昆虫和蜘蛛;72 h 后杭州茶园中捕获10目63科21526个昆虫和蜘蛛.(2)同翅目为优势类群.绍兴茶园中,纯白色板诱捕的同翅目种数占该色板捕获的总物种数的15.2%、同翅目个体数占该色板捕获的总个体数的34.1%;其它9种色板诱捕同翅目种数占各自捕获总种数的21.2%～31.0%、同翅目个体数占各自捕获总个体数的45.8%～75.4%.杭州茶园中,纯白、湖蓝和天蓝色板诱捕同翅目种数小于各自捕获的总种数的20%、同翅目个体数小于各自捕获的总个体数的51%;其它7种色板诱捕同翅目种数大于各自捕获的总种数的22.7%,果绿和墨绿诱捕的同翅目个体数占各自诱捕的总个体数的48.6%和49.8%,桃红、素馨黄、土黄、芽绿和桔黄诱捕的同翅目个体数占各自诱捕的总个体数的55.5%～78.1%.绍兴茶园中,素馨黄和芽绿诱捕的粉虱、假眼小绿叶蝉和广翅蜡蝉分别占两种色板诱捕的总个体数的67.7%和50.1%;杭州茶园中,素馨黄和芽绿诱捕的这几种害虫总数分别占二者诱捕的总个体数的70.7% 和59.2%.(3) 捕获的主要是害虫,绍兴茶园中诱捕的害虫数是天敌的1.63倍,杭州茶园中诱捕的害虫数是天敌的2.23倍.(4)色板之间引诱的种数和个体数差异显著.素馨黄和芽绿诱捕的种数和个体数最多、被捕昆虫的群落多样性指数很小,对叶蝉、粉虱和蜡蝉的引诱力最强;土黄和桔黄诱捕的种数和个体数较多、多样性指数较小,对同翅目昆虫的引诱力较强;纯白色诱捕的种数和个体数居中、多样性指数最大,即诱捕的各种昆虫的个体数差别小;果绿、桃红、墨绿和湖蓝诱捕的种数和个体数明显较少、多样性指数也较小;天蓝色诱捕的种数和个体数最少.(5) 寄生蝇类偏嗜蓝色,缨小蜂类、茧蜂类和小蜂类偏嗜芽绿色.建议将素馨黄和芽绿色板与信息素组合,分别用于监测和诱捕叶蝉和粉虱.     

Glucuronidation of the green tea catechins, (-)-epigallocatechin-3-gallate and (-)-epicatechin-3-gallate, by rat hepatic and intestinal microsomes

The flavonoids (-)-epigallocatechin-3-gallate (EGCg) and (-)-epicatechin-3-gallate (ECg) are major components of green tea and show numerous biological effects. We investigated the glucuronidation of these compounds and of quercetin by microsomes. Quercetin was almost fully glucuronidated by liver microsomes after 3 h, whereas ECg and ECGg were conjugated to a lesser extent ([Formula: See Text] and [Formula: See Text] respectively). The intestinal microsomes also glucuronidated quercetin much more efficiently than ECg and EGCg. Although the rates were lower than quercetin, intestinal microsomes exhibited higher activity on the galloyl group of ECg and EGCg compared to the flavonoid ring, whereas hepatic glucuronidation was higher on the flavonoid ring of EGCg and ECg compared to the galloyl groups. The low glucuronidation rates could partially explain why these flavanols are present in plasma as unconjugated forms.     

Lipid degradation during manufacture of black tea

About 85% of the fatty acids liberated during the manufacture of black tea can be attributed to autolysis of 4 major polar lipid classes in tea leaf tissue, phosphatidylcholine, monogalactosyldiglyceride, digalactosyldiglyceride and phosphatidylethanolamine. Linolenic, linoleic and palmitic acids are the principal fatty acids released from these lipids and they all undergo further degradation. Linolenic acid (60% of fatty acids released) is derived mainly from galactolipids and thus the upper limit of release is dependent on the chloroplast maturity and content of the leaf tissues. Lipid breakdown is complete after 2 hr fermentation and, as there appears to be no accumulation of long chain fatty acid intermediates, it is probable that volatile production has also ceased at this time.