Obtaining of caffeine from Turkish tea fiber and stalk wastes

The aim of this study was to find a cheap method to obtain caffeine. Experiments were performed on fiber and stalk wastes of Turkish tea plants that had no economical value other than being used merely as low grade fuel and fodder. Tea stalks and fiber were obtained from tea factories. Parameters affecting caffeine extraction from tea wastes were determined to be, mixing rate, water/tea ratio, temperature, time and particle size. The maximum yields by dried mass from the tea fibers and stalks were 1.16% and 0.92%, respectively.     

Inhibition of caffeine biosynthesis in tea (Camellia sinensis) and coffee (Coffea arabica) plants by ribavirin

The effects of ribavirin, an inhibitor of inosine-5'-monophosphate (IMP) dehydrogenase, on [8-(14)C]inosine metabolism in tea leaves, coffee leaves and coffee fruits were investigated. Incorporation of radioactivity from [8-(14)C]inosine into purine alkaloids, such as theobromine and caffeine, guanine residues of RNA, and CO(2) was reduced by ribavirin, while incorporation into nucleotides, including IMP and adenine residues of RNA, was increased. The results indicate that inhibition of IMP dehydrogenase by ribavirin inhibits both caffeine and guanine nucleotide biosynthesis in caffeine-forming plants. The use of IMP dehydrogenase-deficient plants as a potential source of good quality caffeine-deficient tea and coffee plants is discussed.     

Identification of novel QTL for black tea quality traits and drought tolerance in tea plants (<Emphasis Type="Italic">Camellia sinensis</Emphasis>)

Tea (Camellia sinensis) contains polyphenols and caffeine which have been found to be of popular interest in tea quality. Tea production relies on well-distributed rainfall which influence tea quality. Phenotypic data for two segregating tea populations TRFK St 504 and TRFK St 524 were collected and used to identify the quantitative trait loci (QTL) influencing tea biochemical and drought stress traits based on a consensus genetic map constructed using the DArTseq platform. The populations comprised 261 F₁ clonal progeny. The map consisted of 15 linkage groups which corresponds to chromosome haploid number of tea plant (2n?=?2×?=?30) and spanned 1260.1 cM with a mean interval of 1.1 cM between markers. A total of 16 phenotypic traits were assessed in the two populations. Both interval and multiple QTL mapping revealed a total of 47 putative QTL in the 15 LGs associated with tea quality and percent relative water content at a significant genome-wide threshold of 5%. In total, six caffeine QTL, 25 catechins QTL, three theaflavins QTL, nine QTL for tea taster score, and three QTL for percent relative water contents were detected. Out of these 47 QTL, 19 QTL were identified for ten traits in three main regions on LG01, LG02, LG04, LG12, LG13, and LG14. The QTL associated with caffeine, individual catechins, and theaflavins were clustered mostly in LG02 and LG04 but in different regions on the map. The explained variance by each QTL in the population ranged from 5.5 to 56.6%, with an average of 9.9%. Identification of QTL that are tightly linked to markers associated with black tea quality coupled with UPLC assay may greatly accelerate development of novel tea cultivars owing to its amenability at seedling stage. In addition, validated molecular markers will contribute greatly to adoption of marker-assisted selection (MAS) for drought tolerance and tea quality improvement.     

Expression and location of caffeine synthase in tea plants

Tea caffeine synthase (CS) is a key enzyme in tea plant for synthesis of caffeine. We firstly systematically investigated the gene expression of CS. Northern blot analysis showed that CS gene was not expressed in stems and roots but efficiently expressed in leaves of tea plants. The expression level of CS gene in summer-grown leaves was much higher than in spring leaves. Its expression level in leaves of the shaded or fertilized tea plants was significantly higher. Along the same shoot, CS gene was expressed at much higher level in the young leaves (bud and the first leaf) than in the more mature second and third leaves. RNA in situ hybridization indicated that tea CS gene was mainly expressed in the palisade parenchyma and the epicuticle of leaves but less expressed in the spongy parenchyma and the hypoderm. Published in Russian in Fiziologiya Rastenii, 2007, Vol. 54, No. 5, pp. 786–789. The text was submitted by the authors in English.     

<Emphasis Type="Italic">Agrobacterium</Emphasis>-Mediated Silencing of Caffeine Synthesis through Root Transformation in <Emphasis Type="Italic">Camellia sinensis</Emphasis> L.

Tea [Camellia sinensis (L.) O. Kuntze] is a perennial and most popular non-alcoholic caffeine-containing beverage crop. Tea has several constraints for its genetic improvement such as its high polyphenolic content and woody perennial nature. The development of transgenic tea is very difficult, laborious, and time taking process. In tea, regeneration requires minimum 8–12 months. In view of this, attempt has been made in this article to develop a rapid, efficient, and quite economical Agrobacterium-mediated root transformation system for tea. The feasibility of the developed protocol has been documented through silencing caffeine biosynthesis. For this, one-month-old tea seedlings were exposed to fresh wounding at the elongation zone of roots and were inoculated with Agrobacterium tumefaciens cultures carrying a RNAi construct (pFGC1008-CS). The pFGC1008-CS contained 376 bp of caffeine synthase (CS) cDNA fragment in sense and antisense direction with an intron in between. This has made the RNAi construct to produce a hairpin RNA (ihpRNA). The suppressed expression of CS gene and a marked reduction in caffeine and theobromine contents in young shoots of tea seedlings were obtained after root transformation through Agrobacterium infiltration. Such transformation system could be useful for functional analysis of genes in tea like woody and perennial plants.     

三峡地区茶园土壤化学特征与茶叶品质的关系

 根据土壤地球化学分类原则,长江三峡地区主要植茶土壤为硅铝质铁铝土、钾硅质铁铝土和硅铁质铁铝土3种土壤地球化学类型。硅铝质铁铝土的特点是土壤质地粗,剖面分异明显,硅、铝、钾含量高,铁、钙、镁含量低：钾硅质铁铝土多为砂壤质,硅、钾含量较高;硅铁质铁铝土土层深厚,质地粘重,铁、铝含量高,磷、钾含量低。土壤地球化学特征与茶叶品质的关系至为密切。砂质土有利于提高茶叶氨基酸含量,而粘质土则会使氨基酸含量降低;较高的盐基含量会降低茶叶茶多酚、咖啡碱和水浸出物含量,但有利于氨基酸形成;速效磷与水浸出物成显著正相关,但与咖啡碱成显著负相关;缓效钾与氨基酸成正相关。从总体上看,硅铝质铁铝土上产出的茶叶品质较高,适宜植茶,硅铁质铁铝土上茶叶品质欠佳,如无特殊改良措施,不宜开辟为茶园。     

Metabolism of methylamine in the tea plant (Thea sinensis L.)

1. The metabolism of methylamine in excised shoot tips of tea was studied with micromolar amounts of [(14)C]methylamine. Of the [(14)C]methylamine supplied 57% was utilized by tea shoots during the 10h experimental period. 2. The main products of [(14)C]methylamine metabolism in tea shoots were serine, gamma-glutamylmethylamide, theobromine, caffeine and CO(2). There was also incorporation of the label into glutamate, aspartate, RNA purine nucleotides and S-adenosylmethionine. 3. The formation of methylamine from gamma-glutamylmethylamide was confirmed by feeding tea shoots with gamma-glutamyl[(14)C]methylamide. The products of gamma-glutamyl[(14)C]methylamide metabolism in tea plants were serine, theobromine, caffeine, glutamate and aspartate. 4. The results indicate that the oxidation of methylamine to formaldehyde is the first step of methylamine utilization. Labelled formaldehyde released by the metabolism of methylamine leads to the incorporation of the label into metabolites on the C(1) pathways of this compound. It is also suggested that formaldehyde is further oxidized via formate to CO(2). 5. The role of gamma-glutamylmethylamide in methylamine metabolism in tea plants is discussed. 6. Results support the view that theobromine is the immediate precursor of caffeine.     

茶树CsPLK基因的鉴定与表达分析

茶树中富含茶氨酸、儿茶素和咖啡碱等重要功能成分,具有较高的价值功效,茶树在生命周期中经常遭受逆境胁迫,维生素B6(VB6)在植物体内参与逆境应答,吡哆醛激酶(pyridoxal kinase,PLK)是VB6补救途径中的关键酶。为进一步了解PLK在茶树生物合成中的功能和作用机理,该研究基于茶树基因组数据库,以龙井43为材料,采用逆转录PCR(RT-PCR)的方法从茶树中克隆出CsPLK的基因。结果表明:该基因序列长为1 179 bp,编码393个氨基酸; CsPLK蛋白和已知物种中PLK蛋白具有较高的同源性,都是核糖激酶超家族成员;通过构建pET-CsPLK载体进行原核表达,并鉴定出重组蛋白有很强的催化活性;组织表达特异性分析表明,叶中的表达量比茎、根的高,在根中最低;荧光定量PCR表示,低温诱导CsPLK上调表达,干旱诱导CsPLK下调表达,发现该基因在茶树中有明显的逆境应答,推测CsPLK在茶树的生长发育、逆境胁迫发挥重要作用。     

贵州苦丁茶植物资源及化学成分分析

苦丁茶是我国南方各民族长期饮用的一类植物代用茶。在贵州,苦丁茶原植物有６科６属９种,主流品种为木樨科女贞属植物粗壮女贞〔Ｌｉｇｕｓｔｒｕｍｒｏｂｕｓｔｕｍ（Ｒｏｘｂ．）Ｂｌ．〕。分析了贵州５种苦丁茶植物叶的主要化学成分并和绿茶进行了对比,分析结果表明,苦丁茶类植物是一种营养成分含量丰富,不含咖啡因的营养保健的功能性饮料新资源     

茶树花挥发物对叶蝉三棒缨小蜂的引诱增强效应

茶树(Camellia sinensis)是我国重要的经济作物。小贯小绿叶蝉(Empoasca onukii)是为害茶树的主要害虫之一。叶蝉三棒缨小蜂(Stethynium empoascae)是小贯小绿叶蝉的主要卵寄生蜂,但茶树花对叶蝉三棒缨小蜂的引诱机制尚不明确。基于析因设计的方差分析结果显示,茶树花可引诱天敌叶蝉...     

茶叶中咖啡因和茶多酚提取技术研究

本文研究茶叶品种和质量、提取剂种类和萃取溶剂量对咖啡因和茶多酚提取得率、提取率和纯度的影响,结果表明:茶叶质量高,咖啡因和茶多酚的含量、得率和纯度就高。95％乙醇提取咖啡因和茶多酚的得率、提取率和纯度最高。1％氧化钙水溶液单纯提取咖啡因的得率、提取率和纯度较高。增加萃取次数和萃取溶剂量可提高咖啡因和茶多酚的得率,对产品纯度没有影响。     

Depression by a green tea extract of alcohol-induced oxidative stress and lipogenesis in rat liver

We determined the effects of a green tea extract with 36% alcohol on the blood alcohol content, oxidative stress, lipogenesis, inflammation and liver function of female Wistar rats. Tea alcohol significantly decreased the O??, H?O? and HOCl amounts via catechins and not caffeine. Thirty days of alcohol gavage improved the level of reactive oxygen species (ROS) in the liver, bile and blood, increased the 4-hydroxynonenal-protein adducts, Kupffer cell infiltration and lipid accumulation in the liver, and elevated the plasma alanine aminotransferase level. A western blot analysis showed reduced expression of the oxidative enzymes (CYP2E1 and NADPH oxidase p47phox protein) and lipogenic enzymes (SREBP-1c and fatty acid synthase) in the alcohol-treated liver. Tea alcohol significantly attenuated these elevated parameters. We conclude that the green tea extract in alcohol efficiently reduced the amounts of O??, H?O? and HOCl primarily due to the catechin content, and not caffeine. The developed tea liquor attenuated alcohol-induced oxidative injury and lipogenesis in the liver by the synergetic action of catechins and caffeine.     

球孢白僵菌对油桐尺蛾的生物效力（英文）

分别在2005年和2006年的5、6月份,在Kamalpur和Hunterpara茶园、孟加拉西部以及印度地区对一种昆虫致病真菌球孢白僵菌对油桐尺蛾Buzura (=Biston) suppressaria Guen.的生物效力进行了评估。以农场主常用的化学杀虫剂40% SP灭多虫和25% EC氯氰菊酯作为标准检测物。试验中球孢白僵菌浓度为1.50 g,2.00 g和2.5 g/lit. of water;灭多虫和氯氰菊酯浓度分别为0.75 mL,1.00 mL和1.50 mL,和0.50 mL,1.00 mL和1.50 mL/lit. of water。在喷洒2.5 g/lit. of water 球孢白僵菌3天后,Kamalpur和Hunterpara茶园中油桐尺蛾种群个体数分别降低了88.00%和84.00%。死的毛虫变成黑色,垂悬在叶片上。并且发现球孢白僵菌的杀虫活性与高剂量的灭多虫和氯氰菊酯的相当。     

Theacrine (1,3,7,9-tetramethyluric acid) synthesis in leaves of a Chinese tea,kucha (Camellia assamica var. kucha)

Theacrine (1,3,7,9-tetramethyluric acid) and caffeine were the major purine alkaloids in the leaves of an unusual Chinese tea known as kucha (Camellia assamica var. kucha). Endogenous levels of theacrine and caffeine in expanding buds and young leaves were ca. 2.8 and 0.6-2.7% of the dry wt, respectively, but the concentrations were lower in the mature leaves. Radioactivity from S-adenosyl-L-[methyl-14C]methionine was incorporated into theacrine as well as theobromine and caffeine by leaf disks of kucha, indicating that S-adenosyl-L-methionine acts as the methyl donor not only for caffeine biosynthesis but also for theacrine production. [8-14C]Caffeine was converted to theacrine by kucha leaves with highest incorporation occurring in expanding buds. When [8-14C]adenosine, the most effective purine precursor for caffeine biosynthesis in tea (Camellia sinensis), was incubated with young kucha leaves for 24 h, up to 1% of total radioactivity was recovered in theacrine. However, pulse-chase experiments with [8-14C]adenosine demonstrated much more extensive incorporation of label into caffeine than theacrine, possibly because of dilution of [14C]caffeine produced by the large endogenous caffeine pool. These results indicate that in kucha leaves theacrine is synthesized from caffeine in what is probably a three-step pathway with 1,3,7-methyluric acid acting an intermediate. This is a first demonstration that theacrine is synthesized from adenosine via caffeine.     

Stimulatory effect of oral administration of green tea and caffeine on locomotor activity in SKH-1 mice

Administration of green tea or caffeine was shown previously to inhibit ultraviolet B light-induced carcinogenesis in SKH-1 mice, and this effect was associated with a reduction in dermal fat. In the present study, oral administration of 0.6% green tea (6 mg tea solids/ml) or 0.04% caffeine (0.4 mg/ml; equivalent to the amount of caffeine in 0.6% green tea) as the sole source of drinking fluid to SKH-1 mice for 15 weeks increased total 24 hr locomotor activity by 47 and 24%, respectively (p<0.0001). Oral administration of 0.6% decaffeinated green tea (6 mg tea solids/ml) for 15 weeks increased locomotor activity by 9% (p<0.05). The small increase in locomotor activity observed in mice treated with decaffeinated green tea may have resulted from the small amounts of caffeine still remaining in decaffeinated green tea solutions (0.047 mg/ml). The stimulatory effects of orally administered green tea and caffeine on locomotor activity were paralleled by a 38 and 23% increase, respectively, in the dermal muscle layer thickness. In addition, treatment of the mice with 0.6% green tea or 0.04% caffeine for 15 weeks decreased the weight of the parametrial fat pad by 29 and 43%, respectively, and the thickness of the dermal fat layer was decreased by 51 and 47%, respectively. These results indicate that oral administration of green tea or caffeine to SKH-1 mice increases locomotor activity and muscle mass and decreases fat stores. The stimulatory effect of green tea and caffeine administration on locomotor activity described here may contribute to the effects of green tea and caffeine to decrease fat stores and to inhibit carcinogenesis induced by UVB in SKH-1 mice.     

Metabolism of xanthine and hypoxanthine in the tea plant (Thea sinensis L.).

1. The metabolism of xanthine and hypoxanthine in excised shoot tips of tea was studied with micromolar amounts of [2(-14)C]xanthine or [8(-14)C]hypoxanthine. Almost all of the radioactive compounds supplied were utilized by tea shoot tips by 30 h after their uptake. 2. The main products of [2(-14)C]xanthine and [8(-14)C]hypoxanthine metabolism in tea shoots were urea, allantoin and allantoic acid. There was also incorporation of the label into theobromine, caffeine and RNA purine nucleotides. 3. The results indicate that tea plants can catabolize purine bases by the same pathways as animals. It is also suggested that tea plants have the ability to snythesize purine nucleotides from glycine by the pathways of purine biosynthesis de novo and from hypoxanthine and xanthine by the pathway of purine salvage. 4. The results of incorporation of more radioactivity from [8(-14)C]hypoxanthine than from [2(-14)C]xanthine into RNA purine nucleotides and caffeine suggest that hypoxanthine is a more effective precursor of caffeine biosynthesis than xanthine. The formation of caffeine from hypoxanthine is a result of nucleotide synthesis via the pathway of purine salvage.     

福建茶树的气候带划分

根据福建山地的气候生态特征和茶树生物学特性,以茶树安全生育的气象指标、茶叶产量形成的热量指标和优质茶形成的关键气象因子为原则,运用气温场计算方程上机计算,结果是:(1)大叶种茶树:闽西南海拔400~560m,闽中海拔200~400m为最适宜带;闽西南海拔600~874m,闽中和闽东北海拔200~600m为适宜带。(2)中、小叶种茶树:闽西南海拔600~874m,闽中、闽东北海拔200~600m为最适宜带;闽西南海拔1200m,闽中海拔1100m,闽北、闽东北海拔600~900m为适宜带;闽西南海拔1500~1700m,闽中海拔1200~1500m,闽北、闽东北海拔950~1200m为次适宜带。据此,作者提出科学开发福建山地,发展茶叶的三条配套技术措施。     

Inhibitors in tea of intestinal absorption of phenylalanine in rats

This work studies the effect of tea extract on the mucosal and serosal transport of phenylalanine, and attempts to identify the active ingredient(s) therein by studying the effect of known tea constituents like theophylline, caffeine and tannic acid. Tea and all the constituents tested inhibited the mucosal uptake of phenylalanine. The serosal transport was unaffected by caffeine and tannic acid, but inhibited by theophylline and high concentrations of tea. The in vitro activity of the intestinal Na⁺-K⁺ ATPase was also assayed from a jejunal homogenate in presence of theophylline, caffeine, tannic acid and cAMP. All were found to inhibit significantly the enzyme. The in vitro activity of a purified Na⁺-K⁺ ATPase was however stimulated by theophylline and caffeine, and inhibited only by tannic acid. It was concluded that the inhibitory effect of tea is exerted mainly through its constituents which inhibit the Na⁺-K⁺ pump directly (tannic acid) or indirectly (theophylline and caffeine), possibly by elevating cAMP levels, dissipating thus the sodium gradient needed for the mucosal uptake of the amino acid.