Polyphenols as potential indicators for drought tolerance in tea (Camellia sinensis L.)

Plant polyphenols have gained prominence in quality of plant products and in human health. An experiment was conducted to determine the association of tea polyphenols with water stress and their suitability as indicators for drought tolerance. The experiment was conducted in a 'rain-out' shelter, and consisted of six tea clones (BBK 35, TRFK 6/8, TRFK 76/1, TRFK 395/2, TRFK 31/30, and TRFK 311/287) and four levels of soil water contents (38, 30, 22, and 14% v/v), which were maintained for a period of 12 weeks. The treatments were arranged in a completely randomized design and replicated three times. Plant growth was monitored over 6 weeks, and a water stress index was calculated to determine water-stress tolerant clones. Total polyphenols in tea shoots was analyzed and a regression analysis done. The results indicate that declining soil water content (SWC) reduced both growth and content of polyphenols in tea. Tolerant clones maintained a high polyphenol content at low SWC, and also showed less fluctuation in phenolics when subjected to changes in SWC. There was significant (P<0.001) correlation of total polyphenol content with shoot growth and WSI of tea, and a linear relationship (r2=0.97) between SWC for tea and both, water stress index and shoot polyphenol content. We report that there is a potential to use polyphenols as indicators for selection of drought-tolerant tea cultivars.     

Shoot epicatechin and epigallocatechin contents respond to water stress in tea [Camellia sinensis (L.) O. Kuntze

An experiment was conducted to determine the association of tea catechins to water stress in tea, with the objective of determining their suitability as indicators for predicting drought tolerance in tea (Camellia sinensis). The study consisted of six tea clones (BBK 35, TRFK 6/8, TRFK 76/1, TRFK 395/2, TRFK 31/30, and TRFK 311/287) and four levels of soil water content (38, 30, 22, and 14% v/v), which were arranged in a complete randomized design and replicated 3 times. The treatments were maintained for a period of 12 weeks. Tea shoots were sampled for catechin analysis during the 6th week of water treatment, in which fresh shoots with two leaves and a bud were plucked and steamed for 2 min, and dried at 70 degrees C to constant weight. Subsequently, the samples were ground and analyzed for catechins using an HPLC system. The total catechins showed significant correlation with shoot growth (r=0.65, P=0.006), soil water content (r=0.54, P=0.0066), and water stress index (r=0.67, P=0.0004). The epicatechin (EC) correlated with shoot growth (r=0.58, P=0.0032), soil water content (r=0.62, P=0.0014), and water stress index (r=0.63, P=0.0010). Similarly, epigallocatechin (EGC) correlated with shoot growth (r=0.65, P=0.0006), soil water content (r=0.50, P=0.0133), and water stress index (r=0.60, P=0.0021). However, epigallocatechin gallate (EGCg) and epicatechin gallate (ECG) showed no significant response to changes in soil water content. The shoot contents of EC and EGC in the six clones showed varied responses, with a distinct pattern in the water-stress tolerant clones (TRFK 6/8 and TRFK 31/30). The results suggest a potential use for EC and EGC as indicators in predicting drought tolerance in tea.     

Polyphenol increases in safflower and cucumber seedlings exposed to strong visible light with limited water

To assess effects of the environmental stress on polyphenol compounds (polyphenols) in plants, the polyphenol contents were investigated in the seedlings of safflower (Carthamus tinctrius L.) and cucumber (Cucumis sativus L.) grown under three types of growth conditions: control; light stress, irradiated with strong light in the visible wavelength range; and light/water stress, irradiated with strong visible light with a limited water supply. The total polyphenol contents and the amounts of the major polyphenols, especially luteolin 7-O-glucoside in safflower cotyledons, and luteolin 7-O-glucoside and luteolin in safflower foliage leaves, increased in response to both stresses. The polyphenol increasing effect of light/water stress was clearly observed in safflower compared to cucumber, suggesting that plants that are resistant to these stresses can accumulate substantial amounts of polyphenols compared to the plants which respond weakly to the stresses.     

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.     

Clonal variation of tea [Camellia sinensis (L.) O. Kuntze] in countering water deficiency

Various clones of tea [Camellia sinensis (L.) O. Kuntze] such as TTL-1, TTL-2, TTL-4, TTL-5, TTL-6, UPASI-2 and UPASI-3 planted in the field were subjected to soil moisture stress conditions by withholding irrigation. A control set of the same clones were maintained by watering regularly. The soil water content of the irrigated and non irrigated plants was monitored through the soil moisture status. The extent of effect of drought on tea plants were monitored through various physiological parameters such as shoot weight, leaf water potential, chlorophyll and carotenoid content, chlorophyll fluorescence (Fv/Fm), net photosynthetic rate, transpiration rate, stomatal conductance and biochemical parameters such as extent of proline accumulation and free radical generation. These parameters were studied on the 30 d of non irrigation and on the 5 d during recovery from drought. The plants recovered when re-irrigated after 30 d of non-irrigation, which suggests that permanent wilting did not occur due to non-irrigation up to 30 d. On the 30 d of non-irrigation the clones TTL-1, TTL-6 and UPASI-2 showed lesser reduction of shoot weight, leaf water potential, chlorophyll fluorescence, photosynthetic rate, transpiration rate and stomatal conductance and increased proline and lesser lipid peroxidation as compared to the other clones. From these results it can be concluded that the clones TTL-1, TTL-6 and UPASI-2 are comparatively more drought tolerant than the clones TTL-2, TTL-4, TTL-5 and UPASI-3.     

茶多酚对NASH 大鼠肝脏组织VEGF 及氧化应激的影响

目的:茶多酚对NASH大鼠肝脏组织VEGF及氧化应激的影响。方法:雄性SD大鼠30只,随机分为3组,正常对照组、模型组、茶多酚治疗组。正常组普通饲料喂养,模型组喂高脂饮食,茶多酚治疗组在高脂饮食12周后茶多酚(150mg(/kg.d)灌胃治疗,16周末处死各组大鼠,留取肝脏组织,观察各组大鼠肝组织病理改变,测定其肝脏丙二醛(MDA)含量和超氧化物歧化酶(SOD)活性以及血管内皮生长因子(VEGF)、Ⅰ、Ⅲ型胶原的表达。结果:模型组大鼠肝组织中SOD活性降低而MDA含量以及VEGF、Ⅰ、Ⅲ型胶原表达均明显高于正常组。茶多酚治疗可减轻肝纤维化程度,显著升高肝组织中SOD活性、降低MDA含量以及VEGF、Ⅰ、Ⅲ型胶原表达水平。结论:茶多酚可通过抑制肝纤维化组织VEGF表达,降低肝组织氧化应激水平而发挥抗肝纤维化作用。     

Protection of osteoblastic cells from freeze/thaw cycle-induced oxidative stress by green tea polyphenol

Green tea polyphenol (GTP) together with dimethylsulphoxide (DMSO) were added to a freezing solution of osteoblastic cells (rat calvarial osteoblasts and human osteosarcoma cells) exposed to repeated freeze/thaw cycles (FTC) to induce oxidative stress. When cells were subjected to 3 FTCs, freezing medium containing 10% (v/v) DMSO and 500 μg GTP ml⁻¹ significantly (p < 0.05) suppressed cell detachment and growth inhibition by over 63% and protected cell morphology. Furthermore, the alkaline phosphatase activity of osteoblastic cells was appreciably maintained after 2 and 3 FTCs in this mixture. Polyphenols may thus be of use as a cell cryopreservant and be advantageous in such fields as cell transplantation and tissue engineering.     

茶多酚对慢性酒精中毒大鼠肝损伤的保护作用

目的：建立慢性酒精诱导的成年大鼠肝损伤动物模型,并进行茶多酚的干预,观察茶多酚的干预对慢性酒精诱导的肝损伤大鼠的防护作用及其可能的机制。方法：将36只SD大鼠适应性喂养一周后,随机分为对照组、酒精损伤组和茶多酚干预组（每组12只）。对照组大鼠用0.9%生理盐水按7 g/kg灌胃,酒精组用体积分数56%的红星牌白酒同剂量灌胃,茶多酚干预组在酒精灌胃同时给予0.25 g/kg剂量的茶多酚。每天定时灌胃一次,连续8周。8周后处死大鼠,取内脏脂肪和肝脏组织,以脂体比衡量内脏脂肪含量,以肝体比和油红O染色结果衡量肝脂质沉积,测定超氧化物歧化酶（SOD）活力、丙二醛（MDA）含量、总抗氧化能力（T-AOC）和谷胱甘肽过氧化物酶（GSH-Px）活力等氧化应激指标,测定肝脏组织中脂肪酸转位酶（FAT/CD36）蛋白水平。结果：与对照组相比,酒精损伤组大鼠内脏脂肪含量、SOD/MDA比值、T-AOC和GSH-Px活力显著下降（（P<0.05或P<0.01）,肝体比、FAT/CD36蛋白水平显著提高（P<0.01）,肝细胞中脂滴增加;与酒精损伤组相比,茶多酚干预组大鼠内脏脂肪含量、SOD/MDA比值、T-AOC和GSH-Px活力显著增加（（P<0.05或P<0.01）,肝体比、FAT/CD36蛋白水平显著下降（P<0.01）,肝细胞中脂滴减少。结论：茶多酚干预能改善慢性酒精中毒大鼠肝脏的脂质沉积和氧化应激状态,并伴有肝细胞膜上FAT/CD36表达的减少。     

Cancer chemoprevention by tea polyphenols

Tea is one of the most widely consumed beverages, second only to water. Many experimental researches in laboratory animals demonstrated that tea components had an inhibitory effect on carcinogenesis at a number of organ sites. The inhibitory effects of tea against carcinogenesis have been attributed to the biologic activities of the polyphenol fraction in tea. This review summarizes experimental data on chemopreventive effects of tea polyphenols in various tumor bioassay systems. Many laboratory studies have demonstrated the inhibitory effects of green tea polyphenols, especially (-)-epigallocatechin-3-gallate (EGCG), on carcinogenesis in animals models. The majority of these studies have been conducted in mouse skin tumor models, where tea polyphenols were used either as oral feeding in drinking water or in direct local application. Most studies used 12-O-tetradecanoylphorbol-13-acetate (TPA) or ultraviolet (UV) radiation as the tumor promoter and found anticarcinogenic effects caused by green tea polyphenols. Black tea was also found to be effective, although the activity was weaker than that of green tea in some experiments. Other studies showed that black tea polyphenols-theaflavins exhibited stronger anticarcinogenic activity than did EGCG. Caffeine in tea was also important for tea to prevent tumorigenesis. The molecular mechanisms of the cancer chemopreventive effects of tea polyphenols are not completely understood. They are most likely related to the mechanisms of biochemical actions of tea polyphenols, which include antioxidative activities, modulation of xenobiotic metabolite enzymes and inhibition of tumor promotion. In addition, we have also proposed that tea polyphenols function as cancer chemopreventive agents through modulation of mitotic signal transduction. However, the molecular mechanisms involved in this modulation need further investigation.     

Analysis of tea polyphenols

Tea is the most highly consumed beverage in the world, other than water. However, unlike water, tea contains substantial amounts of polyphenols that have unique biological activities and may be responsible for many of the health benefits of tea. As a result, it is essential to be able to measure the various tea-associated polyphenols. Total polyphenol content is currently measured by using methodology based on reducing activity. Several HPLC systems with detectors that, collectively, have wide ranges in sensitivity have been developed for analysis of individual flavonoids in tea and biological samples, and for theaflavins in tea. Catechins also have been measured in plasma by solid phase extraction, addition of a chromophore, and colorimetric quantification. Except for theaflavins in tea, routine and robust methods for the measurement of polyphenol condensation products (dimers and thearubigens) in tea and biological samples have not been developed. Although in vitro and animal studies suggest substantial metabolism of flavonoids in the gastrointestinal tract, only a single HPLC procedure has been assembled for monitoring the metabolic products of quercetin in urine of human subjects.     

不同栽培模式对茶园生态环境及茶叶品质的影响

对陕南茶区的纯茶园、塑料大棚茶园、松茶间作茶园及林篱茶园4种常见栽培模式的茶园生态环境(包括小气候因子、土壤因子)和茶叶中各项生化指标(包括氨基酸、咖啡碱、茶多酚、酚氨比、茶水浸出物)进行了测定.结果表明:不同栽培模式茶园小气候因子有显著差异.相对于纯茶园,塑料大棚茶园、松茶间作茶园和林篱茶园能更有效地改善光照条件、降低环境温度、提高土壤水分和有机质含量、增强空气湿度,提高土壤养分状况.3种复合式茶园茶叶品质也较纯茶园显著提高,氨基酸和茶水浸出物含量显著高于纯茶园茶叶,而茶多酚含量却显著低于纯茶园茶叶.因此,林茶复合栽培模式和大棚模式茶园的生态环境更有利于茶树的生长和茶叶品质的提高.     

Growth and leaf gas exchange in Populus euphratica across soil water and salinity gradients

Soil water and salinity conditions of the riparian zones along the Tarim River, northwest China, have been undergoing alterations due to water use by human or climate change, which is expected to influence the riparian forest dominated by an old poplar, Populus euphratica. To evaluate the effects of such habitat alterations, we examined photosynthetic and growth performances of P. euphratica seedlings across experimental soil water and salinity gradients. Results indicated that seedlings were limited in their physiological performance, as evidenced by decreases in their height and biomass, and the maximal quantum yield of photosystem II (PSII) photochemistry (F_v/F_m), the effective quantum-use efficiency of PSII (F_v′/F_m′), and photochemical quenching (q_P) under mild (18% soil water content, SWC; 18.3 g kg^?1 soil salt content, SSC) and moderate (13% SWC, 22.5 g kg^?1 SSC) water or salinity stress. However, seedlings had higher root/shoot ratio (R/S), increased nonphotochemical quenching (NPQ), and water-use efficiency (WUE) relative to control under such conditions. Under severe (8% SWC, 27.9 g kg^?1 SSC) water or salinity stress, P. euphratica seedlings had only a fifth of biomass of those under control conditions. It was also associated with damaged PSII and decreases in WUE, the maximal net photosynthetic rate (P _Nmax), light-saturation point (LSP), and apparent quantum yield (_α). Our results suggested that the soil conditions, where P.euphratica seedlings could grow normally, were higher than ～ 13% for SWC, and lower than ～22.5 g kg^?1 for SSC, the values, within the seedlings could acclimate to water or salinity stress by adjusting their R/S ratio, improving WUE to limit water loss, and rising NPQ to dissipate excessive excitation energy. Once SWC was lower than 8% or SCC higher than ～28 g kg^?1, the seedlings suffered from the severe stress.     

Biomass partition,leaf gas exchange and water relations of alfalfa and milkvetch seedlings in response to soil drying

This study compared physiological and growth responses to water stress of two legume species during the seedling stage. Potted alfalfa (Medicago sativa L. cv. Algonquin) and milkvetch (Astragalus adsurgens Pall. cv. Pengyang earlymaturing vetch) seedlings were grown under well-watered [soil water content (SWC) maintained at 14.92% daily] or water-stressed conditions (drying) for 15 days. Net photosynthetic rate (P _N), transpiration rate (E) and stomatal conductance (g _s) of both species decreased parabolically. When SWC decreased to 7.2% and 10.3%, g _s values for alfalfa and milkvetch were significantly different from those of the respective well-watered plants (p<0.05). When SWC decreased to 6.6% for alfalfa and 6.8% for milkvetch, leaf water potentials (ψ_L) were significantly different from those of the well-watered plants (p<0.05). Thus the difference between the SWC thresholds for a nonhydraulic root signal (nHRS) and a hydraulic root signal (HRS) were 0.6% and 3.5% for alfalfa and milkvetch, respectively. Milkvetch had a lower g _s than alfalfa for a given SWC (p<0.05). Although alfalfa seedlings had a higher dry mass (DM) and root:shoot ratio (R/S) than milkvetch in both treatments (p<0.05), we concluded that milkvetch seedlings had greater drought tolerance than alfalfa.     

Growth, Water Content, and Proline Accumulation in Drought-Stressed Callus of Date Palm

This study was conducted to examine the response of date palm (Phoenix dactylifera L., cvs. Barhee and Hillali) calli to water stress. Callus derived from shoot tip explants was inoculated in liquid Murashige and Skoog medium containing 10 mg dm^–3-naphthaleneacetic acid, 1.5 mg dm^–3 2-isopentenyladenine, and 0 to 30 % (m/v) polyethylene glycol (PEG 8000) to examine the effect of water stress. After 2 weeks, callus growth, water content, and proline accumulation were assessed. Increasing water stress caused a progressive reduction in growth as expressed in callus fresh mass, relative growth rate, and index of tolerance. Both genotypes tested followed this general trend, however, cv. Barhee was more tolerant to drought stress than cv. Hillali. Increasing PEG concentration was also associated with a progressive reduction in water content and increased content of endogenous free proline.     

The Method of ABA Application Affects Salt Stress Responses in Resistant and Sensitive Potato Lines

The phytohormone abscisic acid (ABA) has been proposed to act as a mediator in plant responses to a range of stresses, including salt stress. Most studies of ABA response apply ABA as a single dose. This may not resemble the prolonged increasing endogenous ABA levels that can occur in association with slowly increasing salinity stresses in nature or field situations. Salt stress response based on method of ABA application was examined in four potato genotypes of varying salt stress resistance: the sensitive ABA-deficient mutant and its normal sibling, a resistant genotype line 9506, and commercial cultivar ‘Norland’ of moderate resistance. ABA was applied by root drench at 0, 50, 75, or 100 μM concentrations through a single dose, or by slowly increasing multiple ABA doses in a sand-based growing system under greenhouse conditions. Salt tolerance was then evaluated after 2 weeks of exposure to 150–180 mM NaCl stress. The method of ABA application had a marked effect on the responses to salt stress. Plant responses to the method of ABA application were differentiated according to (1) growth rate, (2) root water content, and (3) apparent shoot growth response. Under a single dose, growth rate increased in all genotypes under salt stress, whereas slowly increasing multiple ABA applications generally maintained stable growth rates except in the ABA-deficient mutant where there was an upward growth trend. Percent root water content was elevated only under slowly increasing multiple ABA doses in two genotypes, whereas none of the single-dose treatments induced any change. The single ABA dose enhanced vertical growth, whereas the slowly increasing multiple ABA dose applications enhanced lateral shoot growth. Because exogenous application is still an artificial system, endogenous ABA was supplied through grafting of ABA-deficient mutant scions onto rootstocks with known elevated ABA levels. Multiple exogenous ABA applications as low as 50 μM elicited similar shoot water content responses as grafting treatments without ABA application in the mutant genotype but had no effect on the ABA normal sibling. Shoot dry weight was significantly increased through grafting over all exogenous ABA treatments. Our study further indicates that the method of ABA application regime in itself can alter plant responses under salt stress and that certain application regimes may reflect responses to elevated endogenous levels of ABA.     

Drought tolerance in three hybrid poplar clones submitted to different watering regimes

Aims Poplars grown in North China may experience water-deficient periods in their life cycle. The aim of the present paper was to quantify the response of three clones to different watering regimes and to determine which clone among the three is the best adapted to drought conditions.Methods Three hybrid poplar clones (clone DN-34, R-247 and OP-367) were used in the present experiment. The seedlings of the three clones were grown under four watering regimes: control (well watered, 100% field water capacity (FC)) and three drought treatments (drought stress I, 50% FC; drought stress II, 40% FC; drought stress III, 30% FC). Changes in morphological, physical and biochemical indicators of the three hybrid poplar clones were investigated.Important findings Drought treatment (50%, 40% and 30% FC) decreased net photosynthetic rate (Pn), transpiration rate (Tr), stomatal conductance (gs), shoot height, total biomass and chlorophyll (Chl) content in all the three clones and it increased activities of antioxidant enzymes and free proline content. The highest values of the above-mentioned morphological and physiological parameters were recorded in clone OP-367 under 30% FC, followed by clone DN-34 and R-247. Relative leaf water content (RWC) and stem diameter (sd) markedly declined in clone R-247 and DN-34 under drought stress I, II and III, whereas RWC and sd declined in clone OP-367 only under drought stress II and III. Clone OP-367 had more RWC and sd than DN-34 and R-247. Only the 30% FC induced an increase in the root-to-shoot ratio (rs) and water use efficiency (WUE) in all the three clones. OP-367 was the most efficient clone in water absorption and use, for plants of the clone had the highest values of rs and WUE. Our data demonstrate that among the three clones, OP-367 was better able to maintain photosynthesis and growth and lower the damage caused by drought.     

茶多酚的抗氧化和抑菌活性及其增效剂

论述了茶多酚的抗氧化和抑菌活性机理,其中起重要作用的是茶多酚分子内的多个酚羟基。在抗氧化过程中,其酚羟基具有供氢体的活性,阻断自由基的链式反应。在抑菌过程中,其酚羟基可与蛋白质分子中的氨基或羧基结合,其疏水性的苯环结构也可与蛋白质发生疏水结合,茶多酚与蛋白质之间的这种多点结合作用阻止了细菌的侵染,使其具有抑菌性。作为增效剂,有机酸、β-胡萝卜紊等能够增强茶多酚的抗氧化性能;壳聚糖、食盐等能够增强茶多酚的抑菌活性．维生素类既能够增强茶多酚的抗氧化性能又能增强其抑菌作用。     

Effect of a strong cold storage induced desiccation on metabolic solutes,stock quality and regrowth,in seedlings of two oak species

Bare-root seedlings of pedunculate oak (Quercus robur L.) and northern red oak (Quercus rubra L.) were lifted in January and stored at 1.8°C, at 82% relative humidity, until their fresh weight declined by 33%. Root growth potential (RGP), fine root electrolyte leakage (REL), fine root water content (RWC), shoot tip water content (SWC), starch and metabolic solute contents in root and shoot, were measured just after lifting and after treatment. Survival of treated seedlings was also assessed in a field trial. RWC, SWC, REL, RGP were dramatically affected by desiccation during cold storage. In both species, root soluble carbohydrate level, inositol level and isocitrate level increased, whereas root starch level and shoot soluble carbohydrate level decreased. In northern red oak, treated seedlings had higher root contents of soluble carbohydrates, inositol and proline than in pedunculate oak. Moreover, treatment induced proline accumulation only in northern red oak roots. These differences could explain why field survival of treated seedlings was significantly better in northern red oak than in pedunculate oak.     

Growth and physiology of aspen supplied with different fertilizer addition rates

Variable internal plant nutrient content may confound plant response to environmental stress. Plant nutrient content may be controlled with relative addition rate techniques in solution culture. However, because raising large numbers of plants in flowing solution culture is difficult, we investigated the feasibility of raising plants in soil mix using relative fertilizer additions. Aspen (Populus tremuloides Michx.) clones (216, 259 and 271) planted in pots containing a peat, sand and vermiculite (2:1:1, v/v/v) soil mix were grown with exponentially increasing fertilizer concentrations and harvested periodically to assess growth. Addition rate treatments ranged from 0.01 to 0.05 day^?1. The lag phase of growth, in which plants adjusted to the fertilizer regime, lasted 40 days after which plants entered the experimental period characterized by constant relative growth rates equivalent to applied fertilizer addition rates. Total plant nutrient concentration was (1) unique for each addition rate, (2) linearly related to addition rate and growth rate, and (3) tended to increase at the highest, and decrease at the lowest addition rates. Regardless, the plants appeared to have attained steady-state conditions. Allocation of carbon to roots increased with lower addition rate treatments and was not dependent upon ontogeny. There were no treatment differences in growth response among aspen clones. Yet there were treatment differences in leaf chlorophyll and photosynthesis within the clones. For the 0.05 day^?1 addition rate treatment, chlorophyll, leaf N concentration and photosynthetic rate were strongly correlated with one another, were at a maximum in recently mature leaves, and rapidly declined with leaf age. The rate of decline in these leaf characteristics was slowest in clone 271, consistent with the leaf longevity stress response reported elsewhere. Plant responses from these relative fertilizer addition trials in soil mix agree closely with those run in hydroponics, indicating that steady-state nutrition can be achieved with a technically simple experimental assemblage.     

Protective Effect of Green Tea Extract and Tea Polyphenols against the Cytotoxicity of 1,4-Naphthoquinone in Isolated Rat Hepatocytes

The cytoprotective effect of green tea extract and its phenolic compounds against 1,4-naphthoquinone-induced hepatotoxicity was evaluated in primary cultured rat hepatocytes. After exposure to 1,4-naphthoquinone, lactate dehydrogenase (LDH) leakage and cell viability were both improved by the presence of the tea extract and tea polyphenols. This cytoprotective effect was related to the structure of tea polyphenols, the galloyl group of (–)-epigallocatechin-3-gallate and (–)-epicatechin-3-gallate being particularly effective. The production of liquid peroxidation by 1,4-naphthoquinone was not inhibited by the tea extract nor by tea polyphenol addition. After 2h of incubation, the protein thiol concentration was reduced by 1,4-naphthoquinone, but this reduction was prevented by the tea extract and tea polyphenols. The reduction in protein thiol content of the cells closely paralleled the LDH leakage and loss of cell viability. These results suggest that the mechanism of protection by tea polyphenols against 1,4-naphthoquinone-induced toxicity to rat hepatocytes was due to the maintenance of protein thiol levels.