8个茶树品种生化成分分析及抗性成分的初步鉴定

【目的】筛选和鉴定茶树中与假眼小绿叶蝉Empoasca vitis G?the相关的抗性成分。【方法】采用氨基酸分析仪和超高效液相色谱法分别分析8个茶树品种的氨基酸及其组分、咖啡碱和茶多酚及其组分的含量。采用人工饲料喂饲法对可能的抗性物质进行初步鉴定。【结果】通过对假眼小绿叶蝉抗性水平差异较大的茶树品种的30多种生化物质的分析,筛选出抗、感茶树品种间差异较大的物质,包括茶多酚、天冬氨酸、γ-氨基丁酸、绿原酸和茶氨酸。分别将其作为可变因子设置不同浓度梯度,加入到人工饲料饲养假眼小绿叶蝉,统计5种化学成份不同浓度人工饲料上假眼小绿叶蝉的成活率,初步确定了这5种化学成份对假眼小绿叶蝉成活率的影响。其中,γ-氨基丁酸可能是茶树抗虫物质之一。【结论】为阐明茶树抗叶蝉的化学机理奠定基础。     

不同施氮量对乌龙茶生长和生理的影响

以武夷肉桂为研究对象,研究不同施氮量对乌龙茶幼龄茶树生长和生理的影响。结果表明,幼龄茶树对氮肥的需求不强烈,其新梢生物量、根生物量和总生物量以及茶叶产量随施氮量的增加而下降。茶树新梢全氮、叶绿素、游离氨基酸、茶多酚和咖啡碱的含量随施氮量的增加而增加,而茶树碳氮比随着施氮量增加而下降;但施氮并没有影响茶树总碳含量。老叶叶绿素含量、根全氮和硝态氮含量、新梢总糖含量与施氮量呈二次曲线回归关系,适度施氮促进根对氮的吸收、老叶叶绿素合成和新梢总糖代谢,过度施氮则相反。新梢生物量与其硝态氮含量和游离氨基酸总量显著负相关;根生物量与根碳氮比和新梢咖啡碱含量显著负相关;茎叶生物量和总生物量与根含氮量显著正相关,但与新梢硝态氮和氨基酸含量显著负相关。过度施氮造成茶树生产力下降的主要原因是因为过度施氮极显著提高了茶树氨基酸代谢水平,使用于茶树生长的碳代谢产物（如总糖）减少,进而影响茶树的生长。     

我国代表性茶树种质嘌呤生物碱的鉴定

从国家种质杭州茶树圃选取了403份有代表性的茶树资源,采用HPLC对嘌呤生物碱进行了2年、春秋两季的重复鉴定。结果表明,年度和季节间咖啡碱含量总体稳定,但可可碱含量春季变化显著。93%以上资源的咖啡碱含量为25.0~45.0 mg/g。茶的3个变种间,咖啡碱含量存在显著差异,且阿萨姆茶秋季较春季咖啡碱含量明显增加。来源于不同地区的资源咖啡碱含量存在显著差异,其中云南和广东的资源变异系数和多样性指数最大。从403份资源中筛选出3份高咖啡碱、1份高苦茶碱(低咖啡碱)和2份高可可碱(低咖啡碱)等6份特异资源。     

茶树花提取物的抑菌和美白功效评价

茶树花作为茶树的生殖生长产物,含有丰富的多酚、皂素、多糖、氨基酸和蛋白等多种活性物质,有抑制微生物生长和减少黑色素沉积的护肤作用。本文分析了茶树花及其提取物(Tea flower extract,TFE)的主要活性成分,并研究了其抑菌和美白功效的机理,为相关功效日化产品提供理论依据。研究的主要结果如下:茶树花及茶树花提取物的主成分种类相同,含量有一定差异,提取工艺后,茶树花提取物可溶性糖含量增加17.51%,多酚含量增加25%,氨基酸含量减少66%,皂素含量降低90%;TFE对金黄色葡萄球菌和大肠杆菌均具有显著的抑菌作用,最小抑菌浓度(MIC)分别为4.5 mg/mL和9.0 mg/mL;小鼠B16黑色素瘤细胞的体外美白实验结果表明,TFE可以通过改变细胞形态、抑制细胞增殖、抑制酪氨酸酶活性、减少黑色素生成等途径达到美白效果。     

土壤微量营养元素对武夷肉桂茶品质的影响

对丹岩区和名岩区各5个取样点的武夷肉桂茶树鲜叶生化成分进行分析。结果表明,名岩区肉桂茶树鲜叶中茶多酚、氨基酸、咖啡碱含量总体均高于丹岩区,名岩区的茶树鲜叶具有比丹岩区更优的物质基础。名岩区产的肉桂成品茶的主要生化成分茶多酚、氨基酸和咖啡碱含量亦略高于丹岩区,其他成分差异不显著。测定武夷肉桂茶树鲜叶和成品茶微量营养元素表明,二者在全铜含量上呈显著性相关,且丹岩区肉桂茶树鲜叶的Cu含量显著高于名岩区。对土壤微量营养元素分析表明,丹岩区的Cu、Zn含量显著高于名岩区,而Fe、Mn含量差异不显著。     

茶树光合作用与光质的关系

在相同辐射能下,茶树叶片光合速率高低依次为黄光>红光>绿光>蓝光>紫光;在相同光量子通量密度下,则依次为红光>蓝紫光>黄光>绿光。生长在不同光质光下的茶树叶片,其光合速率有很大差别,比叶重和气孔密度与光合作用无明显联系。茶叶品质成分(如茶多酚、氨基酸和咖啡碱等)也因光质不同而异。     

蒲公英植酸对沙门氏菌抑制作用及其抑菌机理研究

为探究蒲公英植酸对沙门氏菌的抑制作用及其抑菌机理。本文利用沉淀法和离子交换法提取蒲公英植酸,滤纸片法分析蒲公英植酸对沙门氏菌(Salmonella)的抑菌作用,倍比稀释法研究蒲公英植酸的最低抑菌浓度。通过分析沙门氏菌的细胞通透性和生长动力学,结合扫描电镜和荧光显微镜研究了蒲公英植酸对沙门氏菌的抑菌机理,表明蒲公英植酸对沙门氏菌具有很好的抑菌能力,其最小抑菌浓度为0.2 mg/mL。而且植酸对沙门氏菌的抑制作用是通过破坏细胞膜达到抑菌的效果,并且植酸浓度越高,抑菌效果越显著。这表明蒲公英植酸可以有效地抑制沙门氏菌生长,其主要是通过破坏菌体细胞膜完整性,增加细胞薄膜的通透性,使细胞内容物外溢达到抑制细菌生长的目的。     

镇江香醋醋醅抑菌活性及机理的初步分析

【目的】评价镇江香醋醋酸发酵过程中醋醅及其主要酿醋功能微生物的抑菌活性。【方法】采用琼脂扩散法评价醋醅和功能微生物的抑菌效果,并考察加热、蛋白酶解、透析等不同处理对发酵液抑菌活性的影响。【结果】醋醅水提取液及4种酿醋功能微生物(Acetobacter pasteurianus、A.pomorum、Lactobacillus helveticus、L.plantarum)的发酵上清液对4种指示菌具有明显的生长抑制效果。发酵上清液中的抑菌活性物质具有一定耐热性和蛋白酶敏感性,分子量小于8 k D。【结论】镇江香醋醋醅及其所含醋酸杆菌和乳酸杆菌对环境微生物具有生长抑制活性,且抑菌活性物质的种类具有多样性。     

白化和黄化茶树品种绿茶游离氨基酸、儿茶素类及咖啡碱差异分析

为探究以白化和黄化茶树品种鲜叶为原料制成的绿茶滋味品质和代谢物差异,对广德市6个白化品种绿茶(奶白茶)和14个黄化品种绿茶(黄金芽茶)进行感官审评和代谢物分析。结果表明,奶白茶滋味鲜爽而收敛性略弱;黄金芽茶滋味收敛性强而鲜度低于奶白茶。游离氨基酸总量以及呈现鲜味、甜味的游离氨基酸在奶白茶中的含量显著高于黄金芽茶,而贡献收敛性的儿茶素类化合物和没食子酸含量以及呈现苦味的咖啡碱含量在奶白茶中显著低于黄金芽茶。偏最小二乘法判别分析(PLS-DA)表明导致两种绿茶滋味差异的标志性化合物有7种,分别是茶氨酸、精氨酸、谷氨酸、天冬氨酸、表没食子儿茶素没食子酸酯(EGCG)、表儿茶素没食子酸酯和咖啡碱。味觉活性值(Dot)最高的EGCG在黄金芽茶中的呈味贡献显著高于奶白茶。因此,游离氨基酸、儿茶素类化合物、没食子酸和咖啡碱含量差异导致白化和黄化茶树品种绿茶滋味不同。     

沙棘根瘤内生细菌中抑菌促生菌株的筛选和鉴定

【背景】植物内生细菌可产生具有抑菌和促生活性的物质,既能抑制植物病原菌对寄主植物的侵染,也能促进植物的生长。沙棘根瘤内生细菌是根瘤内除共生固氮的弗兰克氏菌外,与沙棘共生的一大类微生物。研究具有抑菌和促生活性的植物内生菌,可为微生物菌肥的研究提供理论基础。【目的】筛选具有优良抑菌和促生活性的沙棘根瘤内生细菌,初步研究其抑菌和促生活性,并对菌株进行鉴定。【方法】采用双层琼脂法、琼脂扩散法、双层平板对峙法、牛津杯法进行沙棘根瘤拮抗性内生细菌的筛选。选取抑菌活性较高的内生细菌,分别采用Salkowski比色法、ChromeAzurolS(CAS)平板检测法和钼锑抗比色法进行产吲哚乙酸、铁载体及溶磷能力的测定。采用发酵液灌根法测定沙棘根瘤内生细菌SR308对黄瓜促生作用的盆栽效果。通过形态和培养特征、生理生化试验及16S rRNA基因序列分析法对菌株TT201和SR308进行鉴定。【结果】从131株沙棘根瘤内生细菌中筛选出9株具有较强抑菌活性的内生细菌,其中菌株TT201抑菌性最佳、抑菌谱广;菌株SR308的促生活性最好,其发酵液对黄瓜的生长具有较强的促进作用。对具有较强抑菌和促生活性的菌株TT201和SR308进行鉴定的结果表明,菌株TT201为侧孢短芽孢杆菌(Brevibacilluslaterosporus),菌株SR308为蕈状芽孢杆菌(Bacillusmycoides)。【结论】获得2株具有优良抑菌和促生活性的沙棘根瘤内生细菌,为进一步开发微生物农药及菌肥提供了资源。     

湖南省茶树害虫地理区划分析

通过湖南省茶树害虫的系统调查,查明湖南省茶树害虫(害螨)共302种,根据其在湖南省各地(市)的分布,编成"0,1"分布数据表,然后进行聚类分析.根据聚类结果,参考地理、气候、茶树的种植历史和种植面积以及其他动物类群方面的研究和前人的工作,把湖南省茶树害虫分成4个区:湘北环湖平原丘岗区,湘中、湘东丘陵区,湘西山区,湘南山地丘陵区,并分别叙述了各区的特点.     

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.     

不同浓度硒酸钠对茶树的生长和生理指标的影响

该文以一年生扦插苗为材料,采用水培试验,研究不同浓度的硒酸钠(0、0.15、0.3、1.5、3、5、8 mg·L^-1)对茶苗的硒积累、植株生长、生理指标和根尖显微结构等参数的影响。结果表明:茶苗的根和新梢中的硒含量与营养液中的硒浓度正相关;随着硒浓度升高,茶苗的鲜重、侧根数量、根系生物量、光合色素含量、根系活力等生长指标,茶多酚、可溶性蛋白、可溶性糖等茶叶质量指标均呈现先升后降趋势;而丙二醛、过氧化氢、脯氨酸含量等抗逆生理指标则呈现先降后升趋势。显微结构分析显示在不同硒浓度处理条件下根尖的显微构造存在差异。低硒浓度(0.15、0.3、1.5 mg·L^-1)处理的茶苗根尖的皮层薄壁细胞饱满、完好,表皮细胞较小;高硒浓度(Se≥3 mg·L^-1)处理的茶苗根尖的皮层薄壁细胞变形或受损,表皮细胞增厚,表现出胁迫反应。上述结果说明硒对茶树具有双重效应,合适浓度(0.3 mg·L^-1)硒对茶树生长和茶叶品质有益,表现为光合作用和根系活力增强,过氧化物和脯氨酸含量降低,生物量增加,茶叶茶多酚含量增加;硒浓度过高(≥3 mg·L^-1)对茶苗的生长和茶叶品质有害,表现为茶苗出现胁迫反应,茶多酚降低。该研究结果为进一步研究硒对茶树的双重作用机制和富硒茶的栽培提供参考。     

Antimicrobial Activities of Tocklai Vegetative Tea Clones

Thirty-one Tocklai vegetative (TV) tea clones contained caffeine and total catechin 44.39 and 227.55 mg/g dry weight of leaves, respectively. The (−)-epigallocatechin gallate (EGCG) was the most abundant (109.60 mg/g) followed by -(−)-epigallocatechin (EGC, 44.54 mg/g), (−)-epicatechin gallate (ECG, 41.74 mg/g), (−)-epicatechin (EC, 27.42 mg/g) and +catechin (4.25 mg/g). Total catechins were highest in TV 20 (509.7 mg/g) and lowest in TV 6 (71.7 mg/g). The tea clones that contain high level of total catechin exhibited the strongest antimicrobial activity. Among caffeine and flavanol compounds, theaflavins (TF) present in black tea possess a similar antimicrobial potency as EC present in fresh leaves, and that the conversion of catechins to TF during fermentation in making black tea tends to alter their antimicrobial activities. The bioactive molecules other than catechins present in tea leaves may also contribute towards antimicrobial activity.     

Fine control of caffeine biosynthesis in tissue cultures of Camellia sinensis

To determine whether caffeine biosynthesis is controlled by the availability of purine precursors and/or methyl-donors, we examined the effect of some purine compounds on purine alkaloid accumulation, using tea callus cultures. No stimulation of caffeine biosynthesis was observed when the calli were cultured with 0.5 mM adenosine, guanosine or hypoxanthine for 3 weeks. However, 0.5 mM paraxanthine doubled the caffeine level relative to controls. Adenosine stimulated the growth of callus and reduced the caffeine concentration 3 months after inoculation. These results indicate that methylation of xanthosine by 7-methylxanthosine synthase is the most plausible rate-limiting step of caffeine biosynthesis; the supply of non-methylated purine precursors or availability of S-adenosyl-l-methionine are not the principal controlling factors of caffeine biosynthesis. Adenosine salvage to adenine nucleotide synthesis may contribute to the growth of tea calli, but not to caffeine biosynthesis.     

Caffeine affects adventitious rooting and causes biochemical changes in the hypocotyl cuttings of mung bean (<Emphasis Type="Italic">Phaseolus aureus</Emphasis> Roxb.)

Caffeine (1,3,7-trimethylxanthine), a purine alkaloid found naturally in over 100 plant species, has recently been viewed as a safe chemical for management of pests including molluscs, slugs, snails, bacteria, and as a bird deterrent. It possesses phytotoxicity against plant species, yet the mechanism of action is lacking. A study was conducted to determine the effect of caffeine on the rooting of hypocotyl cuttings of mung bean (Phaseolus aureus) and the associated biochemical changes. At lower concentrations (<1,000 μM) of caffeine, though rooting potential was not affected, yet there was a significant decrease in the number of roots and root length. At 1,000 μM caffeine, there was a 68% decrease in the number of roots/primordia per cutting, whereas root length decreased by over 80%. However, no root formation occurred at 2,000 μM caffeine. Further investigations into the biochemical processes linked to root formation revealed that caffeine significantly affects protein content, activities of proteases, polyphenol oxidases (PPO) and total endogenous phenolic (EP) content, in the mung bean hypocotyls. A decrease in rooting potential was associated with a drastic reduction in protein content in the lower rooted portion, whereas the specific activity of proteases increased indicating that caffeine affects the protein metabolism. Activity of PPO decreased in response to caffeine, whereas EP content increased significantly indicating its non-utilization and thus less or no root formation. Respiratory ability of rooted tissue, as determined through TTC (2,3,5-triphenyl tetrazolium chloride) reduction, was impaired in response to caffeine indicating an adverse effect on the energy metabolism. The study concludes that caffeine interferes with the root development by impairing protein metabolism, affecting activity of PPO (and thus lignification), and EP content, which are the crucial steps for root formation.     

Caffeine: a well known but little mentioned compound in plant science.

Caffeine, a purine alkaloid, is a key component of many popular drinks, most notably tea and coffee, yet most plant scientists know little about its biochemistry and molecular biology. A gene from tea leaves encoding caffeine synthase, an N-methyltransferase that catalyses the last two steps of caffeine biosynthesis, has been cloned and the recombinant enzyme produced in E. coli. Similar genes have been isolated from coffee leaves but the recombinant protein has a different substrate specificity to the tea enzyme. The cloning of caffeine biosynthesis genes opens up the possibility of using genetic engineering to produce naturally decaffeinated tea and coffee.     

11种植物精油对6种植物病原真菌的抑菌活性研究

为筛选有效的植物杀菌成分,采用菌丝生长法,测定了香茅油、薰衣草油、菊花油、月桂油、柠檬油、广藿香油、肉桂油、天竺葵油、迷迭香油、茶树油、薄荷油对6种植物病原真菌的抑菌活性。发现在2 g/L的浓度下,上述11种精油对6种供试病菌均有明显的抑制作用,其中香茅油、肉桂油、天竺葵油、月桂油、茶树油和薄荷油对6种病原真菌的抑制率均为100%。剂量效应试验表明,肉桂油对灰葡萄孢（Botrytis cinerea）和禾谷镰孢菌（Fusariumgraminearum）的EC50值分别为29.05μg/mL和42.96μg/mL,而天竺葵油对灰葡萄孢（Botrytis cinerea）和禾谷镰孢菌（Fusarium graminearum）的EC50值分别为34.02μg/mL和68.48μg/mL。     

黑刺粉虱危害对黄金芽茶叶生化成分含量及抗氧化酶活力的影响

【目的】明确黑刺粉虱Aleurocanthus spiniferus为害对茶叶生化物质含量及抗氧化酶活性的影响,为精准、安全、高效、绿色防治黑刺粉虱,提高茶叶品质提供参考。【方法】测定黑刺粉虱为害不同级别下黄金芽芽下第3叶中生化成分(茶多酚、可溶性糖、游离氨基酸、咖啡碱、儿茶素)含量及3种抗氧化酶(SOD, POD和CAT)活性的变化。【结果】随着黑刺粉虱为害等级的提高,黄金芽芽下第3叶中茶多酚的含量显著性降低,且以为害级别为Ⅳ级时最低,为18.82%±0.21%,而游离氨基酸、咖啡碱、可溶性糖和儿茶素的含量在不同的为害级别间均无显著性变化。黄金芽芽下第3叶中POD和CAT活力随着黑刺粉虱为害等级的增加先显著升高后很快降低,均以为害等级Ⅰ时活性最高,分别为57.14±3.98和28.05±0.40 U/mg pro。SOD活力呈显著下降趋势,且以为害等级Ⅳ时SOD活力最低,为442.73±10.54 U/mg pro。【结论】黑刺粉虱为害对黄金芽茶叶中与抗性提高相关的生化物质咖啡碱、儿茶素含量均没有显著性影响,与抗逆补偿相关的游离氨基酸和可溶性糖含量也没有显著性变化,但茶多酚含量显著降低,3种抗氧化酶活性随为害等级升高呈现先升高后显著降低的趋势,说明黄金芽对黑刺粉虱的抵抗力较弱,需要加强黄金芽上黑刺粉虱发生的科学管理。     

The prevention of lung cancer induced by a tobacco-specific carcinogen in rodents by green and black Tea

A growing body of evidence from studies in laboratory animals indicates that green tea protects against cancer development at various organ sites. We have previously shown that green tea, administered as drinking water, inhibits lung tumor development in A/J mice treated with 4-(methylnitrosamino)-1-(3-pyridyl)-l-butanone (NNK), a potent nicotine-derived lung carcinogen found in tobacco. The inhibitory effect of green tea has been attributed to its major polyphenolic compound, epigallocatechin gallate (EGCG), and, to a lesser extent, to caffeine. We have also demonstrated that while levels of O6-methylguanine, a critical lesion in NNK lung tumorigenesis, were not affected in lung DNA. However, the levels of 8-hydroxydeoxyguanosine (8-OH-dG), a marker of oxidative DNA damage, were significantly suppressed in mice treated with green tea or EGCG. These studies underscore the importance of the antioxidant activity of green tea and EGCG for their inhibitory activity against lung tumorigenesis. Unlike green tea, the effect of black tea on carcinogenesis has been scarcely studied, even though the worldwide production and consumption of black tea far exceeds that of green tea. The oxidation products found in black tea, thearubigins and theaflavins, also possess antioxidant activity, suggesting that black tea may also inhibit NNK-induced lung tumorigenesis. Indeed, bioassays in A/J mice have shown that black tea given as drinking water retarded the development of lung cancer caused by NNK. However, data on the relationship of black tea consumption with the lung cancer risk in humans are limited and inconclusive. There is a need for additional tumor bioassays in animal models to better examine the protective role of black tea against lung cancer. The development of adenocarcinomas and adenosquamous carcinomas in F344 rats upon chronic administration of NNK provides an important and relevant model for lung carcinogenesis in smokers. Thus far, no information was previously available regarding the effects of tea on this model. We conducted a 2-year lifetime bioassay in F344 rats to determine whether black tea and caffeine are protective against lung tumorigenesis induced by NNK. Our studies in both mice and rats have generated important new data that support green and black tea and caffeine as potential preventive agents against lung cancer, suggesting that a closer examination of the roles of tea and caffeine on lung cancer in smokers may be warranted.