Effect of microbial fermentation on content of statin, GABA, and polyphenols in Pu-Erh tea

Besides cancer prevention, the hypolipidemic effects of tea have been well studied in animals and humans. Recently, statin has been identified in Pu-erh tea extract. Clinical trials have confirmed that statin decreases the incidence of major coronary and cerebrovascular events and this may be due to its hypolipidemic and antiinflammatory effects. Since a good Pu-erh tea needs longer storage (10 years or more) of fermentation to enhance the flavor and fragrance, we screened microorganisms from two Pu-erh teas, 20 and 25 years old. Species of fungi and bacteria strains that contributed to a good taste of Pu-erh tea were isolated. The effect of fermentation was investigated by inoculating fresh tea leaves with individual strains of isolated microorganisms. Results showed that statin, total polyphenol content, and the scavenging activities of alpha,alpha-diphenyl-beta-picrylhydrazyl (DPPH) radicals increased during fermentation. Tea leaves inoculated with Streptomyces bacillaris strain R9 had the highest polyphenol content (3.3 mg/100 g) and scavenging ability to DPPH radicals (92%). Streptomyces cinereus strain Y11 was equally good for polyphenol content but yielded the highest amount of statin (1012 ng/g) after 42 days of fermentation. Interestingly, the statin content of fresh tea leaves fermented with strain R9 or Y11 after 180 days was much higher (4- and 8-fold, respectively) than that of the 25-year-old Pu-erh tea (513 ng/g) as measured by the HPLC method. Similarly, these two strains also increased the content of gamma-aminobutyric acid (GABA) 5.7- and 4.7-fold in tea fermented for 180 days as compared with the fresh leaves (1270 microg/g) and that were higher than that of the Pu-erh tea (4900 microg/g). Taken together, the present results indicate that tea short-term fermented with S. bacillaris or S. cinereus enhances the color and content of statin, GABA, and polyphenols.     

温度对茶发酵叶色泽及茶色素含量的影响

为探究温度对发酵外形、汤色指标和品质成分的影响,并获得快速判别发酵适度的指标,该文以福鼎大白茶品种鲜叶为原料,设置25、30、35、40℃等4个发酵温度,首次检测在制品外形L值(亮度)、a值(红度)、b值(黄度)和△E值(总色差),汤色LC值(总汤色值)、LL值(汤色透亮度)、La值(汤色红度)和Lb值(汤色黄度)等色泽属性,以及TFs(茶黄素)、TRs(茶红素)和TBs(茶褐素)等品质成分的动态变化,建立3类指标的相关性。结果表明,发酵的进行会导致外形色泽L值、b值的下降,a值的增加,汤色LC值、La值和Lb值的先升后降,LL值的下降,各色泽属性的剧变期均发生在前90 min;不同温度比较,低温(25或30℃)有利于外形亮度L值、外形黄度b值、总汤色LC值、汤色黄度Lb值和汤色红度La的值的稳定,高温(35℃)有利于外形红度a值和汤色透亮度LL值的增加。品质成分TFs和TRs均呈先增后降的趋势,然茶红素峰值点出现时间晚于茶黄素,整体上以25℃发酵有利于TFs的形成和累积,而30℃发酵有利于TRs累积。相关性分析表明,TFs和TRs均与TFRB(茶黄素和茶红素之和与茶褐素的比值)呈极显著正相关,TFRB值较TFR值(茶红素与茶黄素的比值)更可有效代表茶黄素和茶红素在叶内的贡献表征;外形色泽属性与汤色色泽属性间呈极显著相关性,TFs与L值、b值、Lb值呈极显著正相关,与LC值、La值呈显著正相关,TRs仅与a值和Lb值呈显著正相关。通过线性回归分析获得了以LL值、Lb值和b值为变量的茶黄素回归模型,R2达到0.804,结合回归模型和相关性结果可以看出外形黄度b值最为关键,可作为判断发酵适度和衡量茶黄素含量的关键指标。     

Factors affecting the levels of tea polyphenols and caffeine in tea leaves

An isocratic HPLC procedure was developed for the simultaneous determination of caffeine and six catechins in tea samples. When 31 commercial teas extracted by boiling water or 75% ethanol were analyzed by HPLC, the levels of (-)-epigallocatechin 3-gallate (EGCG), and total catechins in teas were in the order green tea (old leaves) > green tea (young leaves) and oolong tea > black tea and pu-erh tea. Tea samples extracted by 75% ethanol could yield higher levels of EGCG and total catechins. The contents of caffeine and catechins also have been measured in fresh tea leaves from the Tea Experiment Station in Wen-Shan or Taitung; the old tea leaves contain less caffeine but more EGCG and total catechins than young ones. To compare caffeine and catechins in the same tea but manufactured by different fermentation processes, the level of caffeine in different manufactured teas was in the order black tea > oolong tea > green tea > fresh tea leaf, but the levels of EGCG and total catechins were in the order green tea > oolong tea > fresh tea leaf > black tea. In addition, six commercial tea extracts were used to test the biological functions including hydroxyl radical scavenging, nitric oxide suppressing, and apoptotic effects. The pu-erh tea extracts protected the plasmid DNA from damage by the Fenton reaction as well as the control at a concentration of 100 microg/mL. The nitric oxide suppressing effect of tea extracts was in the order pu-erh tea >/= black tea > green tea > oolong tea. The induction of apoptosis by tea extract has been demonstrated by DNA fragmentation ladder and flow cytometry. It appeared that the ability of tea extracts to induce HL-60 cells apoptosis was in the order green tea > oolong > black tea > pu-erh tea. All tea extracts extracted by 75% ethanol have stronger biological functions than those extracted by boiling water.     

风障对茶园的减风增温效果及对茶树冠层叶片含水率影响

为解决冬季茶园风寒冻害问题,于2008年1月在安徽农业大学茶场建立塑料薄膜风障,测定风障对茶园的减风增温作用及其对茶树冠层叶片含水率的影响。风障的高度为2.0 m,以5×7网格点测定2.8、2.0 m风障高和1.2 m茶树冠层高度3个层次的风速,以4×4网格点测定地表温度、茶树冠层叶片含水率。结果表明,2.8 m高度,风障上方的风速比环境风速增加30%左右,下风方向的减风作用随风速增大而减小,距离风障7 m区域是风速减弱最强的区域,环境风速2.6 m/s时中轴线风速减小13.5%,环境风速1.0 m/s时     

Effect of microbial decomposition of mature leaves on soil pH

The hypothesis that the microbial decomposition of organic matter low in organic nitrogen leads to an increase in soil pH was tested and confirmed in a pot experiment with two different soils into which mature cherry leaves were incorporated.     

Catechin and caffeine content of green tea dietary supplements and correlation with antioxidant capacity

The health benefits associated with tea consumption have resulted in the wide inclusion of green tea extracts in botanical dietary supplements, which are widely consumed as adjuvants for complementary and alternative medicines. Tea contains polyphenols such as catechins or flavan-3-ols including epicatechin, epigallocatechin, epicatechin gallate, and epigallocatechin gallate (EGCG), as well as the alkaloid, caffeine. Polyphenols are antioxidants, and EGCG, due to its high levels, is widely accepted as the major antioxidant in green tea. Therefore, commercial green tea dietary supplements (GTDS) may be chemically standardized to EGCG levels and/or biologically standardized to antioxidant capacity. However, label claims on GTDS may not correlate with actual phytochemical content or antioxidant capacity nor provide information about the presence and levels of caffeine. In the current study, 19 commonly available GTDS were evaluated for catechin and caffeine content (using high-performance liquid chromatography) and for antioxidative activity [using trolox equivalent antioxidant capacity (TEAC) and oxygen radical antioxidant capacity (ORAC) assays]. Product labels varied in the information provided and were inconsistent with actual phytochemical contents. Only seven of the GTDS studied made label claims of caffeine content, 11 made claims of EGCG content, and five specified total polyphenol content. Caffeine, EGCG, and total polyphenol contents in the GTDS varied from 28 to 183, 12-143, and 14-36% tablet or capsule weight, respectively. TEAC and ORAC values for GTDS ranged from 187 to 15340 and from 166 to 13690 mumol Trolox/g for tablet or capsule, respectively. The antioxidant activities for GTDS determined by TEAC and ORAC were well-correlated with each other and with the total polyphenol content. Reliable labeling information and standardized manufacturing practices, based on both chemical standardization and biological assays, are recommended for the quality control of botanical dietary supplements.     

Analysis of glycosidically bound aroma precursors in tea leaves. 3. Change in the glycoside content of tea leaves during the oolong tea manufacturing process.

A direct qualitative and quantitative determination of the glycosides of tea aroma compounds at the four stages of the oolong tea manufacturing process (plucking, solar withering, indoor withering, and oolong tea product) was carried out by a capillary gas chromatographic-mass spectrometric analysis after trifluoroacetyl derivatization of the glycosidic fractions. Sixteen glucosides and primeverosides were identified and quantified in cv. Chin-shin-oolong and cv. Chinhsuan-oolong. A comparison of the glycosides in dried fresh leaves between the two cultivars showed significant differences. During the manufacturing process, the amounts of most of these glycosides increased from the solar-withering stage, reaching the highest level at the final stage of oolong tea production. It was noted that no glycoside decreased in its content during the manufacturing process, this being quite different from the manufacture of black tea. In addition, the contents of these alcoholic aroma compounds in the free aroma concentrate from each cultivar remained almost unchanged or slightly decreased, and they constituted only about 12 and 17% in amount of the whole oolong tea aroma compounds. However, jasmine lactone and indole were markedly higher in the final oolong tea products.     

转Bt基因棉叶对土壤微生物多样性的影响

应用Biolog方法研究了转Bt基因棉粉碎叶腐解对土壤微生物群落结构功能多样性的影响。取腐解10d、25d、40d、55d、70d土样分析土壤微生物群落多样性指数及土壤微生物对聚合物、胺类、氨基酸、糖、羧酸和其他类碳源利用情况。结果表明:在腐解过程中,转Bt基因棉粉碎叶土壤微生物群落丰富度下降,群落多样性显著降低,而群落优势集中性明显提高;转Bt基因棉粉碎叶影响了土壤微生物群落对碳源的利用程度,表现为可显著增加对糖类、胺类和氨基酸类碳源的利用,初期显著降低对羧酸类碳源的利用,对聚合物类和其他类碳源的利用率无显著影响;主成分分析表明转Bt基因棉粉碎叶对土壤微生物群落原有结构功能影响具有持续性。     

Effect of tylosin and tylosin fermentation waste on microbial activity of soil

The effect of tyiosin fermentation waste and pure tyiosin on microbial activity in soil was investigated. The addition of tylosin fermentation waste at concentrations equivalent to 0.4, 1.2 and 2 tonnes ha^?1 in the field stimulated microbial activity as measured by O₂ uptake. CO₂ release and N mineralization. After 10 weeks incubation half of the C in the waste had been released as CO₂ while mineralization of N ranged between 31 and 38%. Tylosin was detected in the leachates, and at 10 weeks between 20 and 32% of the tylosin present in the waste had been leached from the soil. Addition of high concentrations of pure tylosin. equivalent to those which would be found in 4. 20 and 40 tonnes fermentation waste ha^?1 in the field, resulted in a decrease in microbial respiration for 5–7 weeks after the addition of the antibiotic. N mineralization was reduced throughout the experiment, compared with the control. A higher concentration of tylosin was detected in the leachates in this experiment and after 10 weeks the quantity of tylosin detected represented between 16 and 24% of the tylosin added. It is suggested that the full fertilizer potential of large additions of antibiotic waste to soil could be reduced to some extent due to the inhibitory activity of the residual antibiotic.     

Capillary electrophoretic determination of theanine, caffeine, and catechins in fresh tea leaves and oolong tea and their effects on rat neurosphere adhesion and migration

Theanine, caffeine, and catechins in fresh tea leaves and oolong tea were determined by using capillary electrophoresis (CE). CE separated these tea polyphenols from three other tea ingredients, namely, caffeine, theophylline, and theanine, within 8 min. The young leaves (apical bud and the two youngest leaves) were found to be richer in caffeine, (-)-epigallocatechin gallate (EGCg), and (-)-epicatechin gallate (ECg) than old leaves (from 5th to 7th leaves). On the other hand, the old leaves (from 8th to 10th leaves) contained higher levels of theanine, (-)-epigallocatechin (EGC), and (-)-epicatechin (EC). Results from a comparison of fresh young tea and oolong tea compositions indicated oolong tea contained more theanine and catechins than fresh young tea. Furthermore, it was found that the levels of theanine, EGC, and EGCg in young leaves rose markedly with the withering process. Caffeine did not markedly change. However, fully or partially fermented teas (oolong tea or pauchong tea) have a common initial step in the withering process. Fresh tea leaves or oolong tea extract (0.1%, w/v) markedly inhibited neurosphere adhesion, cell migration, and neurite outgrowth in rat neurospheres. Theanine (348 micrograms/mL) and caffeine at high concentration (50 micrograms/mL) did not inhibit neurosphere adhesion or migration activities, but EGCg at 20 micrograms/mL effectively inhibited neurosphere adhesion for 24 h. These results indicated that EGCg might affect neural stem cell survival or differentiation.     

Effect of selenium on increasing the antioxidant activity of tea leaves harvested during the early spring tea producing season

This research was to determine the effect of foliar application of selenium on increasing the antioxidant activity of tea harvested during the early spring tea producing season using a alpha,alpha-diphenyl-beta-picrylhydrazyl (DPPH) radical scavenging method and the linoleic acid system. The results showed that the radical scavenging ability of the tea extracts followed this order during the first 60 min: selenium-enriched tea obtained by fertilization with selenate > BHT > selenium-enriched tea obtained by fertilization with selenite > alpha-tocopherol > regular tea. Se-enriched tea obtained by fertilization with selenate exhibited the highest inhibition percentage of 84.29% at 30 min. Se-enriched tea extracts provided higher hydrogen-donating capabilities than regular tea and contrasts with BHT and alpha-tocopherol at the concentration of 100 microg of solids/mL of ethanol. There was a little change in the sequence of radical scavenging ability during the later 60 min: Se-enriched tea obtained by fertilization with selenate > Se-enriched tea obtained by fertilization with selenite > BHT > regular tea > alpha-tocopherol. The individual activity of tea extracts and references measured by the linoleic acid system showed that the tea extracts, BHT, and alpha-tocopherol manifested almost the same patterns of activity as the DPPH method. Tea enriched in selenium by fertilization with selenate still exhibited the highest inhibition activity of lipid oxidation, whereas alpha-tocopherol showed the lowest inhibition. The antioxidant activity of Se-enriched green tea harvested during the early spring tea producing season is enhanced compared to regular tea.     

萌芽前叶面喷施硒酸钠提高茶叶含硒量及品质

【目的】探究在茶树不同生育时期叶面喷施不同硒肥对夏茶产量、品质及硒含量的影响,为生产富硒茶提供技术依据。【方法】田间试验在浙江嵊州进行,供试茶树品种为‘中茶108’。试验采用二因素列区设计,主处理为硒肥种类(A因素),副处理为硒肥喷施时期(B因素)。主处理设喷施清水对照(A0)、硒酸钠(A1)、亚硒酸钠(A2)和酵母硒(A3);副处理设夏茶顶芽萌发前(5月12日,B1)与1芽1叶期(5月20日,B2)两个喷施时期。硒肥喷施浓度均为Se 50 mg/L,硒肥溶液喷施量为1.8 L/m2。当茶树蓬面1芽2叶占比达30%左右时,每个小区随机选取30 cm×30 cm茶蓬,调查蓬面新梢总数、1芽2叶数量、1芽2叶长度和百芽重。同时,取1芽2叶新梢样品,测定茶多酚、儿茶素、咖啡碱、游离氨基酸、花青素以及硒含量。【结果】与A0B1处理相比,A3B1处理茶树萌展值显著降低了0.14,但对茶树蓬面新梢总数无明显影响;A2B1处理茶树1芽2叶新梢长度和百芽重分别显著降低了1.04 cm和1.94 g。A1B2和A3B2处理茶树蓬面新梢总数分别显著降低了17.66和22.33,但不影响其萌展值;A1B2显...     

Effect of selenium on the yield and quality of green tea leaves harvested in early spring

Foliar applications of a fertilizer of selenite or selenate were carried out to determine the influence of selenium on the yield and quality of green tea leaves harvested in early spring. Numbers of sprouts and the yield were significantly increased by the application of selenium. The sweetness and aroma of green tea leaves were also significantly enhanced, and bitterness was significantly decreased by the application of selenium. However, no significant differences were found in sweetness, bitterness, and aroma between tea leaves fertilized with selenite and selenate. Se concentration was significantly increased by selenium fertilization, and tea enriched by sodium selenate had a significantly higher selenium content than did tea enriched by sodium selenite. Total amino acid and vitamin C contents were significantly enhanced by the application of selenium. Tea polyphenol contents were significantly decreased by fertilization with selenium. The marked difference of tea polyphenols was also found between applications of selenite and selenate.     

Effect of a commercial plant growth regulator on the elemental content of tea plants

Abstract

Response of field grown mature tea to the foliar application of Biozyme Crop Plus, a commercial plant growth regulator, applied at different concentrations ranging from 250 to 1000 ppm was evaluated. Tea shoots were analysed to determine their N, P, K, Ca, and Mg contents. The treated tea plant shoots contained significantly higher contents of N and K followed by Ca and Mg than the untreated control plant shoots. The total chlorophyll content of Biozyme treated tea shoots was also increased significantly. Yield response to Biozyme applied to tea bushes were also determined.     

不同培肥模式对茶园土壤微生物活性和群落结构的影响

以闽东地区红黄壤茶园定位实验地为对象,通过测定6种不同施肥处理土壤微生物学特性,研究不同培肥对土壤微生物特性和生物化学过程的影响,阐明各指标间的相互关系.结果表明,除了单施无机肥处理外,半量化肥+半量有机肥、全量有机肥、全量化肥+豆科绿肥以及半量化肥+半量有机肥+豆科绿肥等的培肥方式均不同程度提高了土壤有机质,可培养微生物数量,微生物量碳、氮含量及土壤酶活性,尤以半量无机肥+半量有机肥+豆科牧草的培肥模式增幅更为明显,而单施无机肥不利于微生物的生长、酶活性的提高和维持生态系统的稳定性.微生物群落磷脂脂肪酸(PLFAs)标记主成分分析显示,各种不同施肥方式使微生物群落结构发生改变.相关分析表明,微生物量与可培养微生物数量、微生物磷脂脂肪酸含量之间的相关性明显高于微生物量与各种酶活性之间的相关性,说明微生物数量大小对微生物群落结构的影响大于对酶活性功能的影响.研究也表明土壤各微生物指标能从不同方面反映土壤肥力水平,所以采用各种不同的方法能更客观地评价闽东地区茶园红黄壤质量的优劣.     

氮、镁对银杏叶黄酮含量的影响

应用二元二次饱和Ｄ－最优设计进行田间试验,探讨氮、镁对银杏叶黄酮含量的影响。结果表明,在镁缺乏的红土上种植银杏,在合理施用一定的氮肥基础上,增施适量的镁肥,对银杏叶黄酮含量的提高有良好效果。计算机寻优得到供试条件银杏叶黄酮含量最佳施肥方案为：Ｎ５６０ｇ／株,Ｍｇ１５０ｇ／株,目标黄酮含量为２．４８％。     

长期不同施肥对棕壤微生物量磷及其周转的影响

研究棕壤定位试验27年后,长期不同施肥条件下土壤微生物量磷在玉米生长季内的变化及其对植物营养的贡献。结果表明,长期施用化学磷肥或有机肥均能增加土壤微生物磷的含量,尤以有机肥的作用更显著;长期单一的施用氮肥降低了微生物量磷的含量。玉米生长季内土壤微生物量磷的动态变化呈先上升后下降的趋势,其含量最大值出现在玉米生长中后期;一个生长季后,各处理微生物量磷的含量都较施肥前有所下降。长期施肥增加了土壤微生物体的供磷量,微生物体供磷量与玉米产量及吸磷量关系密切,占玉米植株体吸磷量的11.79%～34.46%。不同施肥处理土壤微生物量磷的周转期为0.68～1.61年,施肥延长了微生物量磷的周转期;但单施氮肥加速了其周转。土壤微生物量磷能反映土壤磷素肥力水平。     

施氮对茶园土壤氟和茶树新梢氟含量的影响

通过田间试验研究了不施肥(CK)、施氮360 kg?hm?2(T1)、施氮720 kg?hm?2(T2)处理下茶园土壤无机氮、p H、各形态氟含量的动态变化和春、夏、秋茶树新梢一芽四叶、一芽五叶氟含量,探讨茶园施氮对土壤和茶树新梢氟含量的影响。结果表明:1)茶园施氮后短期内(20~30 d)土壤水溶态氟含量显著降低,土壤交换态氟和铁锰结合态氟含量降低;长期(45~50 d)土壤水溶态氟含量的降低作用减弱,土壤交换态氟和铁锰结合态的含量增加;在试验结束时(164 d),与CK处理相比,T1处理0~20 cm土壤各形态氟含量降低,T2处理0~20 cm土壤各形态氟含量增加。2)0~20 cm茶园土壤水溶态氟、铁锰结合态氟与NH4+-N分别呈极显著负、正相关(P0.01),20~40 cm土壤水溶态氟、交换态氟与NO3?-N分别呈极显著正、负相关(P0.01)。土壤p H与土壤水溶态氟含量极显著负相关(P0.01),与其他3种形态氟含量相关性不显著。土壤铁锰结合态氟与交换态氟、有机结合态氟呈显著、极显著正相关,但与土壤水溶态氟均无显著相关性。3)春茶前后施氮可以降低春、夏、秋茶树新梢一芽四叶、一芽五叶氟含量,但未达显著水平。T1处理新梢氟含量的降低值为夏茶(25.15~27.95 mg?kg?1)秋茶(21.06~24.31 mg?kg?1)春茶(18.58~21.03 mg?kg?1),T2处理的降低值为秋茶(18.64~22.34 mg?kg?1)夏茶(7.79~14.14 mg?kg?1)春茶(3.52~7.30 mg?kg?1)。春、夏、秋茶树新梢氟含量主要受0~20 cm土壤无机氮和20~40 cm土壤p H的影响。因此推测施氮通过影响茶树根系氟的吸收和氟在叶片中的累积过程调控茶树新梢氟含量,该研究成果为合理利用施氮技术降低茶园土壤和茶树新梢氟含量提供了理论依据。     

基于数据融合策略的红茶发酵程度判别

发酵是红茶加工过程中关键的一道工序,对红茶的品质形成有着重要影响。该研究以大叶种英德红茶中的英红九号为研究对象,试验收集了204份不同发酵时间的红茶样品并使用便携式近红外光谱仪和工业相机获取红茶发酵中的信息,基于近红外光谱数据、图像数据和数据融合策略分别建立了红茶发酵程度判别模型。通过分析茶多酚和儿茶素类含量的变化,将红茶的发酵划分为3个阶段,即发酵不足、发酵适度和发酵过度。采用Savitzky-Golay光滑对原始光谱进行预处理,利用竞争自适应重加权采样（Competitive Adaptive Reweighted Sampling, CARS）、连续投影算法（Successive Projections Algorithm, SPA）和主成分分析（Principal Components Analysis, PCA）对近红外光谱变量进行降维处理;相应地,图像进行去阴影后提取了9个颜色特征变量,采用皮尔森（Pearson）相关分析和主成分分析进行特征变量提取。最后采用线性判别分析（Linear Discriminant Analysis, LDA）和支持向量机（Support Vector Machine, SVM）分别建立了基于近红外、图像和两者数据融合的分类模型。结果表明,在建模数据相同的条件下,非线性的支持向量机模型性能优于线性判别分析模型。单一传感器数据建模效果不佳,近红外光谱和图像判别模型的预测集最大准确率仅为83.82%和73.53%。低层次数据融合建模效果较单一传感器数据建模无明显提升,而中层次的数据融合建模效果比单一数据建模均有显著提高,其中SPA提取光谱变量结合Pearson提取图像变量建立的判别模型效果较佳,校正集和预测集准确率分别达到了97.06%和95.59%。研究表明,近红外光谱和视觉结合的中层次融合策略可以作为一种快速判别红茶发酵程度的方法,研究结果为红茶发酵程度构建等级模型与判别奠定了一定的理论基础,为红茶发酵的自动化检测提供了重要依据。