Identification and quantification of main flavonoids in the leaves of Bambusa multiplex cv. Fernleaf

Abstract

The chemical profile of Bambusa multiplex cv. Fernleaf (B. multiplex) leaves was analysed by UPLC-DAD-Q-TOF-MS. Twelve compounds were identified and C-glycosyl flavonoids, including vitexin, isovitexin, isoorientin and its derivatives, are the main constitutes of the plant. Besides, a HPLC method for isoorientin quantification was developed. The RSD of retention time and peak area were 0.05% and 2.04% for six times analysis of isoorientin with concentration of 20?μg/mL. The recovery of isoorientin in real sample was 99.2%. The general trend of isoorientin content in B. multiplex leaves was that it steady increased from Jan. to May, and then quickly decreased. The maximum was found on May with value of 4.7?mg/g. The lowest level of isoorientin was found during Aug. to Nov. with value of about 1.66?mg/g. In different seasons, isoorientin is always the most dominant flavonoid which was accounted for about 50% of total flavonoids in the sample.     

Irradiation application for color removal and purification of green tea leaves extract

Gamma irradiation was introduced to develop a new processing method for brighter-colored green tea leaves extract without changes of physiological activities. Dried green tea leaves were purchased and extracted by 70% ethanol solution and irradiated at 0, 5, 10, and 20 kGy with gamma rays. Hunter color L-value increased and a- and b-value decreased by irradiation, resulting in bright yellow from dark brown. There was no difference in radical scavenging and tyrosinase inhibition effect by irradiation. The irradiation effect in the solution disappeared during storage for 3 weeks at room temperature but vitamin C addition was effective in reducing the color change. Results indicated that irradiation may be a good technology to remove undesirable color in green tea leaves extract.     

The influence of ultraviolet-B radiation on growth, hydroxycinnamic acids and flavonoids of Deschampsia antarctica during Springtime ozone depletion in Antarctica

We examined the influence of solar ultraviolet-B radiation (UV-B; 280-320 nm) on the growth, biomass production and phenylpropanoid concentrations of Deschampsia antarctica during the springtime ozone depletion season at Palmer Station, along the Antarctic Peninsula. Treatments involved placing filters on frames over potted plants that reduced levels of biologically effective UV-B either by 83% (reduced UV-B) or by 12% (near-ambient UV-B) over the 63 day experiment (7 November 1998-8 January 1999) when ozone depletion averaged 17%. Plants growing under near-ambient UV-B had 41% and 40% lower relative growth rates and net assimilation rates, respectively, than those under reduced UV-B. The former plants produced 50% less total biomass as a result of having 47% less aboveground biomass. The reduction in aboveground biomass was a result of a 29% lower leaf elongation rate resulting in shorter leaves and 59% less total leaf area in plants grown under reduced UV-B. p-Coumaric, caffeic and ferulic acids were the major hydroxycinnamic acids, and luteolin derivatives were the major flavonoids in both insoluble and soluble leaf extracts. Concentrations of insoluble p-coumaric and caffeic acid and soluble ferulic acids were 38%, 48% and 60% higher, respectively, under near-ambient UV-B than under reduced UV-B. There were no UV-B effects on concentrations of insoluble or soluble flavonoids.     

INFLUENCE OF PHOTON FLUX DENSITY IN THE 400–700 nm WAVEBAND ON INHIBITION OF PHOTOSYNTHESIS BY UV-B (280–320 nm) IRRADIATION IN SOYBEAN LEAVES: SEPARATION OF INDIRECT AND IMMEDIATE EFFECTS

Abstract— Visible radiation can substantially influence the degree to which plant photosynthesis is inhibited by UV-B radiation. This study was designed to separate the immediate effects of visible radiation on UV-B photosynthetic inhibition from the indirect influence of visible irradiation on morphological and physiological properties of leaves during leaf development. Soybean plants were pretreated in growth chambers with either high or low visible irradiance (750 and 70 μmol m^-2s^-1 quantum flux in the 400–700 nm waveband, respectively) during the development of leaves used subsequently for UV irradiation. Test leaves still attached to the plant were exposed to 5 h of polychromatic UV-B irradiation and the photosynthetic capacity (net CO₂ exchange) was determined before and after the UV irradiation. During the UV irradiation, plants from both pretreatment groups received either high or low visible flux. Development of leaves in the high visible flux pretreatment conditions resulted in thicker leaves, higher chlorophyll a/b ratios, more UV-absorbing pigments, and reduced sensitivity to the UV-B irradiation. However, higher visible flux during the UV-B irradiation resulted in greater depression of photosynthesis by the UV-B irradiation. The relative magnitude of photosynthetic depression under these treatment combinations was the same when photosynthesis was measured under either light-limited or light-saturated conditions.     

Isovitexin-2'-O-beta-[6-O-E-p-coumaroylglucopyranoside] from UV-B irradiated leaves of rice, Oryza sativa L. inhibits fertility of Helicoverpa armigera

UV-B irradiated rice leaves (Oryza sativa L.) contained four closely related flavonoids, with either an isoorientin or isovitexin aglycone. These flavonoids have previously been purified and characterized, and were added to artificial diets of the African bollworm (Helicoverpa armigera Hübner) at 0.1x concentration found in irradiated rice leaves. Consumption of different diets had relatively small effects on laval, pupal and adult duration, weight and survival, indicating the insects lived near normal life cycles on all diets. However, one of the compounds, flavonoid IIa, isovitexin-2'-O-beta-[6-O-E-p-coumaroylglucopyranoside], dramatically reduced the number of fertile eggs laid to 7% of control insects (P<0.001) when added to insect diets at 18 nmol gFW(-1) (14 ppm). A similar antifertility effect was observed when only the male partner consumed diet containing flavonoid IIa, indicating that the reduced fertility may be male specific. In contrast, the fecundity and fertility of insects eating diets containing the closely related flavonoids, isoorientin-2'-O-beta-[6-O-E-p-coumaroylglucopyranoside] or isoorientin-2'-O-beta-[6-O-E-p-feruloylglucopyranoside], were not significantly different to control diets.     

Alteration of foliar flavonoid chemistry induced by enhanced UV-B radiation in field-grown Pinus ponderosa, Quercus rubra and Pseudotsuga menziesii

Chromatographic analyses of foliage from several tree species illustrate the species-specific effects of UV-B radiation on both quantity and composition of foliar flavonoids. Pinus ponderosa, Quercus rubra and Pseudotsuga menziesii were field-grown under modulated ambient (1x) and enhanced (2x) biologically effective UV-B radiation. Foliage was harvested seasonally over a 3-year period, extracted, purified and the flavonoid fraction applied to a mu Bondapak/C(18) column HPLC system sampling at 254 nm. Total flavonoid concentrations in Quercus rubra foliage were more than twice (leaf area basis) that of the other species; Pseudotsuga menziesii foliage had intermediate levels and P. ponderosa had the lowest concentrations of total flavonoids. No statistically significant UV-B radiation-induced effects were found in total foliar flavonoid concentrations for any species; however, concentrations of specific compounds within each species exhibited significant treatment effects. Higher (but statistically insignificant) levels of flavonoids were induced by UV-B irradiation in 1- and 2-year-old P. ponderosa foliage. Total flavonoid concentrations in 2-year-old needles increased by 50% (1x ambient UV-B radiation) or 70% (2x ambient UV-B radiation) from that of 1-year-old tissue. Foliar flavonoids of Q. rubra under enhanced UV-B radiation tended to shift from early-eluting compounds to less polar flavonoids eluting later. There were no clear patterns of UV-B radiation effects on 1-year-old P. menziesii foliage. However, 2-year-old tissue had slightly higher foliar flavonoids under the 2x UV-B radiation treatment compared to ambient levels. Results suggest that enhanced UV-B radiation will alter foliar flavonoid composition and concentrations in forest tree species, which could impact tissue protection, and ultimately, competition, herbivory or litter decomposition.     

Responses of epidermal phenolic compounds to light acclimation: in vivo qualitative and quantitative assessment using chlorophyll fluorescence excitation spectra in leaves of three woody species

Chlorophyll fluorescence (ChlF) excitation spectra were measured to assess the UV-sunscreen compounds accumulated in fully expanded leaves of three woody species belonging to different chemotaxons, (i.e. Morus nigra L., Prunus mahaleb L. and Lagerstroemia indica L.), grown in different light microclimates. The logarithm of the ratio of ChlF excitation spectra (logFER) between two leaves acclimated to different light microclimates was used to assess the difference in epidermal absorbance (EAbs). EAbs increased with increasing solar irradiance intercepted for the three species. This epidermal localisation of UV-absorbers was confirmed by the removal of the epidermis. It was possible to simulate EAbs as a linear combination of major phenolic compounds (Phen) identified in leaf methanol extracts by HPLC-DAD. Under UV-free radiation conditions, shaded leaves of M. nigra accumulated chlorogenic acid. Hydroxybenzoic acid (HBA) derivatives and hydroxycinnamic acid (HCA) derivatives greatly increased with increasing PAR irradiance under the low UV-B conditions found in the greenhouse. These traits were also observed for the HCA of the two other species. Flavonoid (FLAV) accumulation started under low UV-A irradiance, and became maximal in the adaxial epidermis of sun-exposed leaves outdoors. A decrease in the amount of HCA was observed concomitantly to the intense accumulation of FLAV for both leaf sides of the three species. Judging from the logFER, under low UV-B conditions, larger amounts of HCA are present in the epidermis in comparison to FLAV for the three species. Upon transition from the greenhouse to full sunlight outdoors, there was a decrease in leaf-soluble HCA that paralleled FLAV accumulation in reaction to increasing solar UV-B radiation in the three species. In M. nigra, that contains large amounts of HCA, the logFER analysis showed that this decrease occurred in the adaxial epidermis, whereas the abaxial epidermis, which is protected from direct UV-B radiation, continued to accumulate large amounts of HCA.     

Changes of microsomal membrane properties in spring wheat leaves (Triticum aestivum L.) exposed to enhanced ultraviolet-B radiation

The properties of microsomal membranes in spring wheat leaves (Triticum aestivum L. cv. Ganlong No. 92-005) exposed to (0) control, 8.64 (T1) and 11.2 kJ m(-2) day(-1) (T2) biologically effective UV-B irradiation (UV-B(BE)) were studied under greenhouse conditions. These irradiance levels correspond to a decrease in the stratospheric ozone of approximately 12.5 and 20%, respectively, for a clear solstice day at Lanzhou (36.04 degrees N, 1550 m), China. Compared with controls, the content of malondialdehyde (MDA) increased by 70.8% in T1 and 83.8% in T2 on the 7th day of the radiation, and the IUFA (index of unsaturated fatty acids) decreased, indicating peroxidation of lipid acids. Simultaneously, a drastic decrease of phospholipid content after 21 days and an increase of membrane lipid microviscosity on UV-B irradiation were also found, suggesting a reduction in the fluidity of membrane lipids. Ethylene emission by the microsomal membrane, in the presence of exogenous 1-aminocyclopropane-1-carboxylic acid was higher in the wheat seedlings after 7, 14 and 21 days' irradiation than in the controls. These changes were correlated with a rise in lipoxygenase activity. Membrane-bound enzymes (Ca2+ -ATPase and Mg2+ -ATPase) were promoted by UV radiation in the first 7 days and significantly decreased after 14 and 21 days' treatment in comparison to control. Our results suggest that UV-B radiation may cause changes in structural complexity and function of microsomal membranes in spring wheat leaves.     

Flavonoid concentrations in three grass species and a sedge grown in the field and under controlled environment conditions in response to enhanced UV-B radiation

An investigation was carried out to find whether enhanced levels of UV-B radiation induce increased concentrations of flavonoids in the leaves of the grass species Deschampsia antarctica, Deschampsia borealis and Calamagrostis epigeios and the sedge Carex arenaria. Whether the enhanced levels of UV-B influenced the proportions of the various flavonoids in the leaves was also studied. Increased flavonoid concentrations would improve the UV-B shielding of UV-B susceptible tissues. Using HPLC analysis the flavonoids orientin and luteolin were identified in D. antarctica, orientin in D. borealis and tricin in C. arenaria. Neither flavonoid concentrations nor the proportion of the various flavonoids in climate room grown D. antarctica and D. borealis plants differed between individuals grown under 0, ambient or elevated UV-B levels. After 12 weeks of growth biomass production and shoot-to-root ratios of D. antarctica were not affected by elevated UV-B radiation. Greenhouse grown C. epigeios plants contained higher concentrations and different proportions of flavonoids grown under elevated levels of UV-B than when grown under ambient or 0 UV-B. In C. epigeios plants grown in their natural habitat in the field under ambient or elevated levels of UV-B, flavonoid concentrations and proportions were the same in plants from both treatments. In the leaves of the sedge C. arenaria grown in a greenhouse flavonoid concentrations and proportions were not affected by UV-B radiation. Leaves were harvested four times during the growing season from C. arenaria plants grown in their natural habitat in the field under ambient or elevated levels of UV-B. Leaves harvested in January contained higher concentrations of flavonoids when grown under elevated UV-B than when grown under ambient UV-B radiation. In leaves harvested in May, September and December flavonoid concentrations were the same in plants grown under ambient or elevated UV-B. The proportion of the different flavonoids was the same for both treatments in all months. These results indicate that constitutive levels of flavonoids in these grass and sedge species are adequately high to protect them against ambient and elevated levels of solar UV-B radiation.     

UV-B RADIATION EFFECTS ON PHOTOSYNTHESIS, GROWTH and CANNABINOID PRODUCTION OF TWO Cannabis sativa CHEMOTYPES

The effects of UV-B radiation on photosynthesis, growth and cannabinoid production of two greenhouse-grown C. sativa chemotypes (drug and fiber) were assessed. Terminal meristems of vegetative and reproductive tissues were irradiated for 40 days at a daily dose of 0, 6.7 or 13.4 kJ m^-2 biologically effective UV-B radiation. Infrared gas analysis was used to measure the physiological response of mature leaves, whereas gas-liquid chromatography was used to determine the concentration of cannabinoids in leaf and floral tissue.
There were no significant physiological or morphological differences among UV-B treatments in either drug- or fiber-type plants. The concentration of Δ⁹-tetrahydrocannabinol (Δ⁹-THC), but not of other cannabinoids, in both leaf and floral tissues increased with UV-B dose in drug-type plants. None of the cannabinoids in fiber-type plants were affected by UV-B radiation.
The increased levels of Δ⁹-THC in leaves after irradiation may account for the physiological and morphological tolerance to UV-B radiation in the drug-type plants. However, fiber plants showed no comparable change in the level of cannabidiol (a cannabinoid with UV-B absorptive characteristics similar to Δ⁹ THC). Thus the contribution of cannabinoids as selective UV-B filters in C. sativa is equivocal.     

Dose–Response and Temperature Dependence of the Mortality of Spider Mite and Predatory Mite Eggs Caused by Daily Nighttime Ultraviolet-B Irradiation

The two-spotted spider mite, Tetranychus urticae, is an economically important agricultural pest. A novel physical control method involving daily nighttime UV-B irradiation was recently developed for use in strawberry greenhouses. However, the overlapping of leaves after March prevents direct irradiation to T. urticae on the lower leaf surface, decreasing control effect. Excessive UV-B irradiation causes leaf sunscald in winter. Therefore, optimization of UV-B irradiance and a compensatory control agent are desired. Temperature may affect the survival of organisms exposed to UV-B, although the temperature dependence of UV-B damage is controversial. A phytoseiid mite, Neoseiulus californicus, is a prominent predator but vulnerable to a single UV-B irradiation. We compared dose–response and temperature dependence of UV-B damage between T. urticae and N. californicus eggs under daily nighttime UV-B irradiation. Unexpectedly, N. californicus showed greater resistance to UV-B than T. urticae, and the mortality was increased and decreased at low and high temperatures, respectively. This makes possible the application of UV-B doses that are lethal for spider mites but safe for phytoseiid mites. Overall, we concluded that combined use of phytoseiid mites with UV-B lamps is advantageous to spider mite management in strawberry greenhouses.     

Determination of Anthocyanins and Flavonols in Paeonia delavayi by High-Performance Liquid Chromatography with Diode Array and Mass Spectrometric Detection

The colors of the leaves in Paeonia delavayi may be pure green, green-red, or dark red according to their habitat. The goal of this study was to compare the type and the total concentrations of anthocyanins and flavonols in red and green P. delavayi. The constituents and concentration of anthocyanins and flavonols were identified and determined by high-performance liquid chromatography with diode array detection and mass spectrometry. Cyanidin-3, 5-di-O-glucoside, cyanidin-3-O-glucoside, and peonidin-3-O-glucoside were first determined in red P. delavayi. There were no anthocyanins in green P. delavayi. Seven flavonols were identified in both red and green P. delavayi with differences in their concentrations. The concentrations of total anthocyanins and flavonoids were determined at 525 and 360?nm using the external standard method. The total anthocyanins in red P. delavayi were 152.24?mg/100?g in leaves and 78.92?mg/100?g in stems. The total flavonoids were 805.4?mg/100?g in leaves and 438.3?mg/100?g in stems, which were much higher than in green P. delavayi for both leaves and stems. This study reports composition and concentration differences in red and green P. delavayi based on ecological habitat.     

Solar ultraviolet-B radiation increases phenolic content and ferric reducing antioxidant power in Avena sativa

We examined the influence of solar ultraviolet-B radiation (UV-B; 280-320 nm) on the maximum photochemical efficiency of photosystem II (F(v)/F(m)), bulk-soluble phenolic concentrations, ferric-reducing antioxidant power (FRAP) and growth of Avena sativa. Treatments involved placing filters on frames over potted plants that reduced levels of biologically effective UV-B by either 71% (reduced UV-B) or by 19% (near-ambient UV-B) over the 52 day experiment (04 July-25 August 2002). Plants growing under near-ambient UV-B had 38% less total biomass than those under reduced UV-B. The reduction in biomass was mainly the result of a 24% lower leaf elongation rate, resulting in shorter leaves and less total leaf area than plants under reduced UV-B. In addition, plants growing under near-ambient UV-B had up to 17% lower F(v)/F(m) values early in the experiment, and this effect declined with plant age. Concentrations of bulk-soluble phenolics and FRAP values were 17 and 24% higher under near-ambient UV-B than under reduced UV-B, respectively. There was a positive relationship between bulk-soluble phenolic concentrations and FRAP values. There were no UV-B effects on concentrations of carotenoids (carotenes + xanthophylls).     

RESPONSE DIFFERENCES BETWEEN TWO SOYBEAN CULTIVARS WITH CONTRASTING UV-B RADIATION SENSITIVITIES

Abstract— Soybeans (Glycine max [L.] Men. cvs. Essex and Williams) were grown in an unshaded greenhouse under two levels of biologically effective ultraviolet-B (UV-B_BE) radiation (effective daily dose: 0 and 11.5 kJ m^-2) for 34 days. Ultraviolet-B radiation reduced leaf area and total plant mass in Essex but these parameters were unaffected in Williams. Differences in both anatomical and biochemical characteristics were found between cultivars. Some of these differences were inherently distinct between cultivars while others were variably induced by UV treatment. Specific leaf weight. an estimate of leaf thickness, was unchanged in Essex but increased in Williams with UV-B irradiation. The relative increase in concentration of UV-absorbing compounds in leaf tissues after UV-B irradiation was greater in Williams. The composition of UV-absorbing compounds in leaf tissues differed between the two cultivars but was unaffected by UV-B radiation. Although total soluble proteins and total peroxidase activity were similar between cultivars, several electrophoretically distinct peroxidase activities were detected. Therefore, the intraspecific variation in UV-B sensitivity found in soybean appears to be correlated with a suite of anatomical and biochemical differences, including leaf thickness, composition and concentration of UV-absorbing compounds in leaf tissues, and possibly differences in peroxidase activities.     

Effects of enhanced UV-B on pigment-based phytoplankton biomass and composition of mesocosm-enclosed natural marine communities from three latitudes

A series of three outdoor mesocosm experiments was undertaken in Rimouski (Canada), Ubatuba (Brazil) and Ushuaia (southern Argentina) to examine the effects of lamp-enhanced UV-B (280-320 nm) on phytoplankton communities isolated from seawater at each site. Detailed pigment composition was used to identify these communities. Each experiment compared three replicated UV-B treatments, consisting of natural sunlight conditions (NUVB), low-level UV-B enhancement corresponding to local 30% ozone depletion (LUVB) and high-level enhancement corresponding to 60% ozone depletion (HUVB). Each mesocosm (ca 2 m deep) was mixed continuously (turnover time, ca 1.3 h) and samples were obtained daily over 7-10 days. In Rimouski a large diatom bloom occurred during the first week. Repeated-measures analysis of variance (RM-ANOVA), with time as the repeated factor, showed slight but statistically significant increases in the chlorophyll (Chl) a level with the HUVB treatment, which were especially obvious over the last 3 days of the experiment. A large decrease in grazers (ciliates) that was observed concurrently with this treatment is the most likely explanation for the increase in Chl a level. The lack of negative effect on algal biomass by enhanced UV-B is attributed to the mixing inside the mesocosms and to the relatively low UV-B penetration. In Ubatuba levels of most pigments decreased over time, particularly fucoxanthin, Chl c3 and alloxanthin. The RM-ANOVA showed no effect of the UV-B treatments, except for Chl c3, which had significantly lower concentrations under natural UVB conditions, indicating that enhanced UV-B directly or indirectly favored Chl c3 algae (likely prymnesiophytes). Although particulate organic carbon concentration was significantly larger during HUVB treatment than during the other treatments, Chl a was unaffected, suggesting that enhanced UV-B favored heterotrophs. Lack of algal growth during this experiment was attributed to low nutrient concentrations (which were the lowest of the three sites), high irradiances (which were the highest noon incident photosynthetically available radiation and UV of the three sites) and UV-B penetration down to the bottom of the mesocosms. In Ushuaia a small bloom took place over the first 5 days. The RM-ANOVA showed no overall effect of the UV-B treatments for any of the pigments examined but on the last 3 days of the experiment several green algae-type pigments, such as Chl b and siphonein, showed increased concentrations under the HUVB treatment. UV-B enhancement hence favored green algae, as seen from the stronger increase over time in the ratio of Chl b to Chl a associated with the HUVB treatment. UV-B enhancement also seemed to cause a slight decrease in physiological condition, because the relative concentration of chlorophyllide a and some pheophorbides that may be the product of dying algae increased during the HUVB treatments in Ubatuba and particularly in Ushuaia (where UV-B also penetrated to the bottom of mesocosms). For all three sites changes in phytoplankton biomass due to the UV-B treatments were minor, even though UV-B enhancement was important. This study indicates that effects of enhanced UV-B on the community structure of both phytoplankton and their grazers are potentially more important than effects on overall algal biomass.     

Effects of elevated ultraviolet radiation and endophytic fungi on plant growth and insect feeding in Lolium perenne, Festuca rubra, F. arundinacea and F. pratensis.

Plants of perennial ryegrass (Lolium perenne L.), red fescue (Festuca rubra L.), tall fescue (F. arundinacea Schreb.) and meadow fescue (F. pratensis Huds) were exposed at an outdoor facility located in Edinburgh, UK to modulated levels of UV-B radiation (280-315 nm) using banks of cellulose diacetate filtered UV-B fluorescent lamps that also produce UV-A radiation (315-400 nm). The plants were derived from a single clone of each species and were grown both with and without colonization by naturally-occurring fungal endophytes. The UV-B treatment was a 30% elevation above the ambient erythemally-weighted level of UV-B during July to October. Growth of treated plants was compared with plants grown under elevated UV-A radiation alone produced by banks of polyester filtered lamps and with plants grown at ambient levels of solar radiation under banks of unenergized lamps. At the end of the treatment period, sample leaves were collected for feeding trials with the desert locust Schistocerca gregaria (Forsk). The UV-B treatment produced no effects on the aboveground biomass of any of the four grasses. The UV-B treatment and the UV-A control exposure both increased plant height and the number of daughter plants formed by rhizome growth in F. rubra. There were significant effects of endophyte presence on the total fresh and dry weights of F. arundinacea and F. rubra, on fresh weight only in F. pratensis, and on the fresh and dry weights of inflorescence in F. arundinacea and L. perenne. There were no effects of UV treatments on the absolute amounts of leaf consumed or on the feeding preferences of locusts for leaves with or without endophyte in three species: F. rubra, F. arundinacea and L. perenne. In F. pratensis there was no effect of UV treatment on the weight of leaves consumed but a significant UV x endophyte interaction caused by a marked change in feeding preference between leaves with and without endophyte that differed between the UV-B treatment and UV-A control exposures. The alkaloid compounds known as lolines were analysed in leaves of F. pratensis and were only found in plants grown with endophyte. However, there was no significant relationship between total loline content and insect feeding preference. These effects illustrate the potential complexities of species interactions under increasing levels of UV-B. The experiment also demonstrates the importance of appropriate controls in UV lamp supplementation experiments for interpretation of both plant growth and insect feeding effects.     

UV-B-induced secondary conformational changes in lens alpha-crystallin.

The changes in turbidity and protein secondary structure of alpha-crystallin after a 72 h UV-B (302 nm) irradiation in aqueous solution have been determined by UV spectrophotometry and Fourier transform infrared (FT-IR) microspectroscopy with reflection mode. The relative transmission of alpha-crystallin aqueous solution gradually decreases with the exposure time, indicating that the transparent alpha-crystallin aqueous solution becomes opaque with prolonged UV-B irradiation. The turbidity induced by UV-B shows first-order kinetics due to the photo-induced aggregation. The modification of the secondary structure of the alpha-crystallin molecule in aqueous solution caused by this aggregation might enhance the alpha-helix and beta-turn structures from 8.14 to 14.92% and from 24.46 to 35.54%, respectively; reduce the beta-sheet structure from 60.20% to 43.77%; and leave the random coil structure almost unaltered. The secondary conformation of alpha-crystallin changes gradually but evidently with its increase of turbidity during UV-B exposure.     

Further knowledge on barley (Hordeum vulgare L.) leaves O-glycosyl-C-glycosyl flavones by liquid chromatography-UV diode-array detection-electrospray ionisation mass spectrometry

Thirty-seven flavonoids and a hydroxycynnamic acid have been characterized in barley leaves (Hordeum vulgare L.) by liquid chromatography-UV diode-array coupled to ion trap mass spectrometry with electrospray ionisation interface (negative mode). Their structures have been determined by the study of the ion mass fragmentation which characterizes C-glycosyl flavones and O-glycosyl-C-glycosyl flavones, and differentiates di-O-glycosyl flavones from O-diglycosyl-flavones. The majority of them are described for the first time in barley. Saponarin (isovitexin-7-O-glucoside), lutonarin (isoorientin-7-O-glucoside) and isoscoparin-7-O-glucoside derivatives constitute the major part of the detected compounds. Some acylated derivatives are also described, namely, 7-O-[6-acyl]-glucoside and -7-O-[6-acyl]-glc-4'-glucoside of isovitexin, isoorientin and isoscoparin.     

The effects of grape seeds polyphenols on SKH-1 mice skin irradiated with multiple doses of UV-B

The study investigated the protective activity of red grape seeds (Vitis vinifera L, Burgund Mare variety) (BM) extracts in vivo on multiple doses of ultraviolet radiation (UV)-B-induced deleterious effects in SKH-1 mice skin. Eighty 8-weeks-old female SKH-1 mice were divided into 8 groups: control, vehicle, UV-B irradiated, vehicle+UV-B irradiated, BM 2.5mg polyphenols (PF)/cm(2)+UV-B irradiated, BM 4 mg PF/cm(2)+UV-B irradiated, UV-B+BM 2.5mg PF/cm(2), UV-B+BM 4 mg PF/cm(2). The extract was applied topically before or after each UV-B exposure (240 mJ/cm(2)), for 10 days consecutively. The antioxidant activity of BM extract is higher than gallic acid (k(BM)=0.017, k(gallic acid)=0.013). Multiple doses of UV-B generated the formation of cyclobutane pyrimidine dimers (CPDs) and sunburn cells, increased glutathione peroxidase (GPx) and catalase (CAT) activities respectively glutathione (GSH) and IL-1β levels in skin. In group treated with 2.5mg PF/cm(2) before UV-B irradiation BM extract inhibited UV-B-induced sunburn cells, restored the superoxide dismutase (MnSOD) activity, increased insignificantly CAT and GPx activities and reduced IL-1β level. The BM 4.0 mg PF/cm(2) treatment decreased GSH level and reduced the percentage of CPDs positive cells in skin. Both doses of BM extract administered after UV-B irradiation increased the MnSOD and GPx activities and reduced the formation of sunburn cells in skin. Our results suggest that BM extract might be a potential chemo-preventive candidate in reducing the oxidative stress and apoptosis induced by multiple doses of UV-B in skin.     

Solar and supplemental UV-B radiation effects in lemon peel UV-B-absorbing compound content-seasonal variations

Effects of solar and supplemental UV-B radiation on UV-B-absorbing compounds and malondialdehyde (MDA) accumulations in the peel of lemons collected in summer and winter were analyzed. UV-B-absorbing compounds were higher in flavedo than in albedo tissue in both seasons; however, the highest values were observed in summer. These compounds were also higher in outer than in inner flavedo surface. Lemons were categorized as sun-, semisun- and shaded-lemon according to localization inside the tree canopy. Depending on-tree localization UV-B-absorbing compounds were higher in flavedo of sun-lemon than in semisun- and shaded-lemon. Supplementary UV-B radiation (22 kJ m(-2) day(-1) UV-BBE) induced UV-B-absorbing compound synthesis in on-tree and postharvest lemons. Two minutes of supplemental UV-B irradiation in summer lemons produced a strong increment (300%) of UV-B-absorbing compound content, whereas in winter lemons a slight increase (30%) was observed only after 3 min of irradiation. By contrast, UV-B-absorbing compound accumulation was not observed in albedo. MDA accumulation showed approximately a similar trend of UV-B-absorbing compounds. According to our results, solar UV-B was not required for UV-B-absorbing compound accumulation in lemon peel. Relationships between UV-B-absorbing compounds, MDA, reactive oxygen species and pathogen protection are also discussed.