Impact of solar ultraviolet-B radiation on the antioxidant defense system in soybean lines differing in flavonoid contents

Exposure to ultraviolet-B (UV-B) radiation can lead to oxidative damage in plants. However, plants possess a number of UV-protection mechanisms including screening of potentially damaging UV-B and increased production or activities of antioxidants. The balance or trade-off between these two mechanisms has rarely been studied and is poorly understood. Two isolines of soybean (Glycine max [L.] Merr.) Clark cultivar, the normal line with moderate levels of flavonoids and the magenta line with reduced flavonoids levels, were grown in the field with or without natural levels of UV-B. Leaflet blades of the first trifoliate leaf were harvested after 4–12 days of exposure to the experimental conditions for analysis of active oxygen species (AOS) and antioxidant levels. Solar UV-B radiation caused oxidative stress in both lines and altered AOS metabolism primarily by decreasing superoxide dismutase activity and increasing the activities of ascorbate peroxidase, catalase and glutathione reductase. This resulted in decreased ascorbic acid content and increased dehydroascorbate content. The magenta line had greater oxidative stress than the normal line in spite of its enhanced oxidative defense capacity as compared to the normal line, even under UV-B exclusion. These results indicate enhanced sensitivity in the magenta line, especially under UV-B exclusion that was likely due to the absence of flavonoid epidermal screening compounds and subsequent increased penetration of solar ultraviolet radiation into the leaf.     

滤减UV-B辐射对烤烟蛋白质组变化的影响

为研究不同UV-B辐射强度对烤烟生理代谢及调控途径的影响,应用蛋白质双向电泳联用质谱技术,以云南普遍种植的烤烟K326为试验材料,通过覆盖不同透明薄膜滤减UV-B辐射的方式,对75.8%(聚乙烯膜,处理1)和37.5%(麦拉膜,处理2)UV-B辐射透过率处理下K326的蛋白质组和相关生理性状进行了比较。结果表明:在蛋白质组中有10个蛋白在这两类处理下蛋白差异表达显著;与处理1相比,在处理2的K326叶片中有5个蛋白上调表达,5个蛋白下调表达;通过质谱分析共鉴定出8种功能明确的蛋白质,其中差异表达的10个蛋白中有3个与氧化还原相关,3个与光合作用相关,1个是参与能量代谢的激酶蛋白,1个是RNA结合蛋白,另外还有2个未知功能的蛋白待探明,在蛋白质组水平对不同UV-B辐射强度与烤烟生长发育的关系进行了初步研究;而在K326的生理成熟期、过渡期及工艺成熟期,处理2的净光合速率(Pn)均高于处理1,这与所鉴定出的3个与光合作用有光的蛋白在处理2中上调表达趋势一致;相比之下,处理1的K326发育进程较快,茎粗等形态指标及比叶重均比处理2高。     

短乳杆菌DM9218核苷酸代谢过程中的蛋白组学分析

目的探讨短乳杆菌DM9218在核苷酸代谢过程中的蛋白表达差异。方法分别提取DM9218菌株与底物(肌苷+鸟苷)反应前后的菌体蛋白,利用蛋白双向凝胶电泳(2-DE)技术,找出该菌株与底物反应前后的差异蛋白质点,选取其中差异变化较大的蛋白点进一步做蛋白质谱分析。结果 2-DE分析显示两样品蛋白点主要分布在等电点4~9和分子量11~90 kD范围内,将所得的蛋白点结合其蛋白得率、浓度、储存蛋白含量进行比较,得到匹配的蛋白点数为732个。从中选取14个差异显著的蛋白点进行质谱分析,质谱结果显示所选取蛋白质点主要与物质代谢、能量转换及基因水平转录和翻译等生物学功能密切相关。结论本研究为后期分析研究短乳杆菌DM9218在核苷酸代谢过程中蛋白的表达奠定了基础。     

Ambient levels of UV-B in Hawaii combined with nutrient deficiency decrease photosynthesis in near-isogenic maize lines varying in leaf flavonoids: Flavonoids decrease photoinhibition in plants exposed to UV-B

Near-isogenic lines of maize varying in their genes for flavonoid biosynthesis were utilized to examine the effects of foliar flavonoids and nutrient deficiency on maximum net photosynthetic rate (P _N) and chlorophyll (Chl) fluorescence (F_v/F_m) in response to ultraviolet-B (UV-B) radiation. Plants with deficient (30 to 70 % lower N, K, Mn, Fe, and Zn) and sufficient nutrients were exposed to four irradiation regimes: (1) no UV-B with solar photosynthetically active radiation (PAR), (2) two day shift to ambient artificial UV-B, 8.2–9.5 kJ m⁻² d⁻¹ (21–25 mmol m⁻² d⁻¹); (3) continuous ambient artificial UV-B; (4) continuous solar UV-B in Hawaii 12–18 kJ m⁻² d⁻¹ (32–47 mmol m⁻² d⁻¹). The natural ratio of UVB: PAR (0.25–0.40) was maintained in the UV-B treatments. In the adequately fertilized plants, lines b and lc had higher contents of flavonoids and anthocyanins than did lines hi27 and dta. UV-B induced the accumulation of foliar flavonoids in lines hi27 and b, but not in the low flavonoid line dta or in the high flavonoid line lc. In plants grown on deficient relative to adequate nutrients, flavonoid and anthocyanin contents decreased by 30–40 and 40–50 %, respectively, and Chl a and Chl b contents decreased by 30 and 70 %, respectively. The UV-B treatments did not significantly affect P _N and F_v/F_m in plants grown on sufficient nutrients, except in the low flavonoid lines dta and hi27 in which P _N and F_v/F_m decreased by ∼15 %. P _N, F_v/F_m, and stomatal conductance decreased markedly (20–30 %) in all lines exposed to UV-B when grown on low nutrients. The decrease in F_v/F_m was 10 % less in higher flavonoid lines b and lc. The photosynthetic apparatus of maize readily tolerated ambient UV-B in the tropics when plants were adequately fertilized. In contrast, ambient UV-B combined with nutrient deficiency significantly reduced photosynthesis in this C₄ plant. Nutrient deficiency increased the susceptibility of maize to UV-B-induced photoinhibition in part by decreasing the contents of photoprotective compounds.     

Proteomic analysis of early germs with high-oil and normal inbred lines in maize

High-oil maize as a product of long-term selection provides a unique resource for functional genomics. In this study, the abundant soluble proteins of early developing germs from high-oil and normal lines of maize were compared using two-dimensional gel electrophoresis (2-DGE) in combination with mass spectrometry (MS). More than 1100 protein spots were detected on electrophoresis maps of both high-oil and normal lines by using silver staining method. A total of 83 protein spots showed significant differential expression (>two-fold change; t-test: P < 0.05) between high-oil and normal inbred lines. Twenty-seven protein spots including 25 non-redundant proteins were identified by matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Functional categorization of these proteins was carbohydrate metabolism, cytoskeleton, protein metabolism, stress response, and lipid metabolism. Three such proteins involved in lipid metabolism, namely putative enoyl-ACP reductase (ENR), putative stearoyl-ACP desaturase (SAD) and putative acetyl-CoA C-acyltransferase (ACA), had more abundant expressions in high-oil lines than in normal. At the mRNA expression level, SAD, ENR and ACA were expressed at significantly higher levels in high-oil lines than in normal. The results demonstrated that high expressions of SAD, ENR and ACA might be associated to increasing oil concentration in high-oil maize. This study represents the first proteomic analysis of high-oil maize and contributes to a better understanding of the molecular basis of oil accumulation in high-oil maize.     

Comparative proteome analysis of drought-sensitive and drought-tolerant rapeseed roots and their hybrid F1 line under drought stress

Rapeseed (Brassica napus L.), which is the third leading source of vegetable oil, is sensitive to drought stress during the early vegetative growth stage. To investigate the initial response of rapeseed to drought stress, changes in the protein expression profiles of drought-sensitive (RGS-003) and drought-tolerant lines (SLM-003), and their F1 hybrid, were analyzed using a proteomics approach. Seven-day-old rapeseed seedlings were treated with drought stress by restricting water for 7 days, and proteins were extracted from roots and separated by two-dimensional polyacrylamide gel electrophoresis. In the sensitive rapeseed line, 35 protein spots were differentially expressed under drought stress, and proteins related to metabolism, energy, disease/defense, and transport were decreased. In the tolerant line, 32 protein spots were differentially expressed under drought stress, and proteins involved in metabolism, disease/defense, and transport were increased, while energy-related proteins were decreased. Six protein spots in F1 hybrid were common among expressed proteins in the drought-sensitive and -tolerant lines. Notably, tubulin beta-2 and heat shock protein 70 were decreased in the drought-sensitive line and hybrid F1 plants, while jasmonate-inducible protein and 20S proteasome subunit PAF1 were increased in the F1 hybrids and drought-tolerant line. These results indicate that (1) V-type H⁺ ATPase, plasma-membrane associated cation-binding protein, HSP 90, and elongation factor EF-2 have a role in the drought tolerance of rapeseed; (2) The decreased levels of heat shock protein 70 and tubulin beta-2 in the drought-sensitive and hybrid F1 lines might explain the reduced growth of these lines in drought conditions.     

Age-related changes in the expression and oxidation of bronchoalveolar lavage proteins in the rat

The incidence and severity of many lung diseases change with age. Some diseases, such as pneumonia, occur with increased frequency in children and the elderly. Proteins obtained by bronchoalveolar lavage (BAL) serve as the first line of defense against inhaled toxins and pathogens. Age-related changes in BAL protein expression and oxidative modification were examined in juvenile (1 mo), young adult (2 mo), and aged (18 mo) F344 rats using two-dimensional difference gel electrophoresis (2D-DIGE), matrix-assisted laser desorption ionization-time of flight/time of flight (MALDI-ToF/ToF) tandem mass spectrometry, and carbonyl immunoblotting. Using 2D-DIGE, we detected 563 protein spots, and MALDI-ToF/ToF identified 204 spots comprising 31 proteins; 21 changed significantly (17 increases) between juvenile and young adult or aged rats, but for 12 of these proteins, levels had a biphasic pattern, and levels in aged rats were less than in young adults. Relative carbonylation was determined by comparison of immunostaining with total protein staining on each oxidized protein blot. We found that aged rats had significantly increased oxidation in 13 proteins compared with juvenile rats. Many of the proteins altered in expression or oxidation level had functions in host defense, redox regulation, and protein metabolism. We speculate that low levels of expression of host defense proteins in juvenile rats and decreases in levels of these proteins between young adult and aged rats may predispose these groups to pneumonia. In addition, we have shown age-related increases in protein oxidation that may compromise host defense function in aged rats.     

INHIBITION OF GROWTH OF ULVA EXPANSA (CHLOROPHYTA) BY ULTRAVIOLET-B RADIATION1

We examined the effect of ultraviolet-B radiation (UV-B, 290–320 nm) on the growth rate of the intertidal marine alga Ulva expansa (Setch.) S. & G. (Chlorophyta). Segments of thallus collected from a natural population were grown in outdoor seawater tanks. Combinations of UV-B-opaque screens, UV-B-transparent screens, and UV-B lamps were used to investigate the effects of solar UV-B and solar plus supplemental UV-B on the growth of these segments. Growth was measured by changes in segment surface area, damp weight, and dry weight. Growth rates of segments were inhibited under both solar UV-B and solar plus supplemental UV-B treatments. Growth rates were also inhibited by high levels of photosynthetically active radiation, independent of UV-B fluence. These results indicate that increases in UV-B resulting from further ozone depletion will have a negative impact on the growth of this alga.     

Proteome analysis on an early transformed human bronchial epithelial cell line,BEP2D,after alpha-particle irradiation

To probe the mechanism of carcinogenesis of lung cancer at the molecular level and to find potential protein markers involved in the early phase of tumorgenesis, differential proteome analysis on primary passage cell line R15H, and early transformed cell line R15H20 derived from (238)Pu alpha-particle irradiation of human papillomavirus (HPV) 18-immortalized human bronchial epithelial cell line (BEP2D), was carried out using two-dimensional electrophoresis (2-DE) and peptide mass fingerprinting (PMF) with matrix-assisted laser desorption/ionisation-time of flight mass spectrometry. Image analysis and Student's t-test (p < 0.05) showed that three protein spots were only expressed in R15H, intensities of 43 protein spots on the gels were altered between R15H and R15H20. Two of the three spots that were only expressed in R15H were identified as high mobility group protein 1. Two proteins decreased in abundance in R15H20 were identified as maspin precursor, a tumor suppressor and aminoacylase-1. Ornithine aminotransferase and peptidyl-prolyl cis-trans isomerase A that were increased in R15H20, were also identified. Relationships between these differentially expressed proteins and the carcinogenesis mechanism of lung cancer are discussed. The protein expression profile of the R15H cell line was also constructed during the study as a reference map for further comparative proteome analysis of the irradiation induced BEP2D cell line. Of the 90 spots analyzed with PMF in the 2-DE gel of R15H cell line, 50 proteins were identified by searching the nonredundant protein database SWISS-PROT/TrEMBL.     

Proteome analysis of antibody-producing CHO cell lines with different metabolic profiles

Two-dimensional gel electrophoresis and tandem mass spectrometry were used to identify proteins associated with a metabolic shift during fed-batch cultures of two recombinant antibody-producing CHO cell lines. The first cell line underwent a marked change in lactate metabolism during culture, initially producing lactate and then consuming it, while the second cell line produced lactate for a similar duration but did not later consume it. The first cell line displayed a declining specific antibody productivity during culture, correlating to the 2-D gel results and the intracellular antibody concentration determined by HPLC. Several statistical analysis methods were compared during this work, including a fixed fold-change criterion and t-tests using standard deviations determined in several ways from the raw data and mathematically transformed data. Application of a variance-stabilizing transformation enabled the use of a global empirical standard deviation in the t-tests. Most of the protein spots changing in each cell line did not change significantly in the other cell line. A substantial fraction of the changing proteins were glycolytic enzymes; others included proteins related to antibody production, protein processing, and cell structure. Enolase, pyruvate kinase, BiP/GRP78, and protein disulfide isomerase were found in spots that changed over time in both cell lines, and some protein changes differed from previous reports. These data provide a foundation for future investigation of metabolism in industrially relevant mammalian cell culture processes, and suggest that along with differences between cell types, the proteins expressed in cultures with low lactate concentrations may depend on how those conditions were generated.     

Analysis of nitroso-proteomes in normotensive and severe preeclamptic human placentas

Nitric oxide (NO) plays a key role in placental biology, and placental dysfunction is the main pathogenesis pathway for preeclampsia, yet the direct placental targets of NO actions have not been determined. Covalent adduction of an NO moiety to cysteines, termed S-nitrosylation (SNO), is emerging as a key route by which NO can directly modulate protein functions. This study was conducted to analyze global S-nitroso (SNO)-proteins in human placentas and to determine if their levels differ in normotensive versus severe preeclamptic placentas. Although total nitrite/nitrate increased, total levels of SNO-proteins and nitrosylated forms of endothelial NO synthase and heat shock protein 90 were decreased by preeclampsia. We further compared normotensive and preeclamptic placental nitroso-proteomes (total SNO-protein profiles) by using a biotin and CyDye switch test combined with two-dimensional fluorescence difference gel electrophoresis (2D-DIGE) and identified SNO-proteins by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Numerous SNO-proteins were displayed as spots on 2D-DIGE gels. One hundred spots of interest were excised; 46 spots were identified, of which 8 spots were novel SNO-proteins; levels of 15 spots were increased, and 6 spots were decreased, and the rest were unchanged by preeclampsia. Pathway analysis suggested that placental SNO-proteins are involved in regulating various cellular functions including protein synthesis, cell movement and metabolism, cell signaling, and other functions. These data therefore show for the first time that SNO is a crucial mechanism by which NO directly regulates placental proteins linked to various biological pathways. The significantly altered placental nitroso-proteome in preeclampsia suggests that SNO plays a role in the placental pathophysiology in preeclampsia.     

增强UV-B辐射和氮对谷子叶光合色素及非酶促保护物质的影响

以谷子(Setaria italica(L)Beauv.)为对象,从拔节期开始持续给予低氮(1.875 mmol/L)和高氮(15 mmol/L)两种氮供应条件并从抽穗期开始进行26 d两种强度(4.29、7.12 kJ·m-2·d-1)的增强UV-B辐射处理,研究了谷子叶中光合色素含量、类黄酮含量和苯丙氨酸解氨酶(PAL)活性的变化.结果表明:与高氮供应条件相比,低氮供应条件明显降低了谷子叶中光合色素含量但提高了类胡萝卜素/叶绿素含量比值;在开花期中段和灌浆期中段,高氮供应条件下谷子叶中光合色素含量对增强UV-B辐射比低氮供应条件下的谷子更敏感.从灌浆期开始到处理结束,两种影响因子对谷子叶中类黄酮含量均有显著影响,增强UV-B辐射导致谷子叶中类黄酮含量逐渐升高,且相同增强UV-B辐射强度下低氮供应条件下的谷子叶中类黄酮含量明显高于高氮供应条件下的谷子.谷子叶中PAL活性对两种影响因子的响应较类黄酮含量更加敏感,低氮供应条件使谷子叶中PAL活性明显提高.结合上述指标的相关性分析结果可知,低氮供应条件加强了处于繁殖期主要阶段的谷子叶中类黄酮的积累,并使谷子叶中的类胡萝卜素/叶绿素含量比值明显提高,进而有助于维持谷子叶中光合色素含量在增强UV-B辐射条件下的相对稳定性,对植株抵抗UV-B辐射伤害有利.     

Differential responses of alpine and non-alpine Aquilegia species to increased ultraviolet-B radiation

Abstract. Two species of columbines, Aquilegia caerulea (which lives in alpine environments) and A. canadensis (a lower-elevation species) responded to artifically increased levels of solar UV-B radiation by becoming significantly shorter and developing a significantly greater number of leaves. The degree of shortening was greater in A. canadensis , the non-alpine species, while the number of leaves increased faster in A. caerulea . Significant differences in gashandling and water-use physiological responses were noted, with UV-B treatment leading to decreased water-use efficiency in both species. Concentrations of flavonoids increased significantly in both species following light exposure, whereas alkaloids in A. caerulea appeared to be destroyed by UV-B. No significant changes occurred in β-carotene concentrations in either species.     

Response of purely symbiotic and NO3-fed nodulated plants of Lupinus luteus and Vicia atropurpurea to ultraviolet-B radiation

The effects of elevated UV-B radiation on growth, symbiotic function and concentration of metabolites were assessed in purely symbiotic and NO3-fed nodulated plants of Lupinus luteus and Vicia atropurpurea grown outdoors either on tables under supplemental UV-B radiation or in chambers covered with different types of plexi-glass to attenuate solar ultraviolet radiation. Moderately and highly elevated UV-B exposures simulating 15% and 25% ozone depletion as well as sub- ambient UV-B did not alter organ growth, plant total dry matter and N content per plant in both L. luteus and V. atropurpurea. In contrast, elevated UV-B increased (P <0.05) flavonoid and anthocyanin concentrations in roots and leaves of L. luteus, but not of V. atropurpurea. Feeding nodulated plants of L. luteus under elevated UV-B radiation with 2 mM NO3 increased (P <0.05) nodule, leaf and total dry matter, and whole plant N content. With V. atropurpurea, NO3 reduced (P <0.05) nodule activity, root %N and concentrations of flavonoids, anthocyanins in roots and leaves and soluble sugars in roots, in contrast to an observed increase (P <0.05) in nodule dry matter per plant. Similarly, supplying 2 mM NO3 to L. luteus plants exposed to sub-ambient UV-B radiation significantly reduced individual organ growth, plant total biomass, nodule dry matter, nodule %N, and whole plant N content, as well as root concentrations of flavonoids, anthocyanins, soluble sugars, and starch of L. luteus, but not V. atropurpurea plants. These results show no adverse effect of elevated UV-B radiation on growth and symbiotic function of L. luteus and V. atropurpurea plants. However, NO3 supply promoted growth in L. luteus plants exposed to the highly elevated UV-B radiation.     

Arabidopsis Flavonoid Mutants Are Hypersensitive to UV-B Irradiation

Increases in the terrestrial levels of ultraviolet-B (UV-B) radiation (280 to 320 nm) due to diminished stratospheric ozone have prompted an investigation of the protective mechanisms that contribute to UV-B tolerance in plants. In response to UV-B stress, flowering plants produce a variety of UV-absorptive secondary products derived from phenylalanine. Arabidopsis mutants with defects in the synthesis of these compounds were tested for UV-B sensitivity. The transparent testa-4 (tt4) mutant, which has reduced flavonoids and normal levels of sinapate esters, is more sensitive to UV-B than the wild type when grown under high UV-B irradiance. The tt5 and tt6 mutants, which have reduced levels of UV-absorptive leaf flavonoids and the monocyclic sinapic acid ester phenolic compounds, are highly sensitive to the damaging effects of UV-B radiation. These results demonstrate that both flavonoids and other phenolic compounds play important roles in vivo in plant UV-B protection.     

Proteomic investigation of metabolic shift in mammalian cell culture.

Mammalian cells, under typical cultivation conditions, produce large quantities of lactate and ammonia that affect cell growth adversely and result in low cell concentration. Controlled nutrient feeding to maintain low concentrations of glucose and glutamine reduces metabolite production drastically, altering the metabolism of the cells. This metabolic shift results in higher cell concentration in continuous cultures and does not affect the specific productivity of the cells. We have taken a proteomics approach to investigate the differential protein expression with metabolic shift. Using two-dimensional gel electrophoresis (2-DE) and mass spectrometry (MS), we have found at least eight differentially expressed spots; two proteins were down-regulated, and the others were up-regulated with metabolic shift. These included metabolic enzymes, the brain form of phosphoglycerate mutase, which was down-regulated, and the precursor of the 23 kDa subunit of NADH-ubiquinone oxidoreductase, which was up-regulated. Another enzyme, the L1 isozyme of ubiquitin carboxyl-terminal hydrolase, which is involved in protein turnover and degradation, was also up-regulated in the metabolically altered cells. The remaining down-regulated spot had been identified as two isoforms of cytoplasmic actins, while three of the up-regulated spots were viral GAG polyproteins from various murine viruses. An unidentified protein was also up-regulated in the cells with altered metabolic state. This study shows the potential of using a proteomics approach in deciphering the intracellular changes in cells with physiological changes such as metabolism shift. The new insight into cell metabolism afforded by this analysis will greatly facilitate process optimization of continuous cell cultures.     

Ultraviolet-B radiation impacts light-mediated turnover of the photosystem II reaction center heterodimer in Arabidopsis mutants altered in phenolic metabolism

Ultraviolet-B (UV-B) radiation can have a negative impact on the growth and development of plants. Plants tolerant to UV-B alleviate these effects using UV-screening pigments that reduce the penetration of UV-B into mesophyll tissue. Little is known about the relative contribution of specific phenolic compounds to the screening capacity of leaves. The D1 and D2 proteins constituting the photosystem (PS) II reaction center heterodimer are targets of UV-B radiation and can be used as an in situ sensor for UV penetration into photosynthetic tissue. Degradation of these proteins occurs under very low fluences of UV-B, and is strongly accelerated in the presence of visible light. Using the D1-D2 degradation assay, we characterized UV-B sensitivity of Arabidopsis mutants (tt4, tt5, and fah1) that are genetically altered in their composition of phenolic compounds. We found that changes in phenol metabolism result in altered rates of PSII reaction center heterodimer degradation under mixtures of photosynthetically active radiation and UV-B. A comparison of D2 degradation kinetics showed increased UV sensitivity of the Landsberg (Landsberg erecta) tt5 mutant relative to the Landsberg tt4 mutant and the Landsberg wild type. Despite a lack of flavonoid accumulation, the tt4 mutant is not particularly UV sensitive. However, the tolerance of this mutant to UV-B may reflect the increased accumulation of sinapate esters that strongly absorb in the UV range, and may thus protect the plant against environmentally relevant UV-B radiation. This sinapate-mediated protection is less obvious for the tt4 mutant of Columbia ecotype, indicating that the relative contribution of particular phenolics to the total screening capacity varies with the genetic background. The role of sinapate esters in UV screening is further substantiated by the results with the fah1 mutant where absence of most of the sinapate esters results in a significantly accelerated degradation of D2 under mixed light conditions. Because the latter mutant is not expected to be deficient in flavonoids, the relative contribution of flavonoids as protectants of PSII reaction center heterodimer against UV-B damage in Arabidopsis needs to be re-evaluated vis-a-vis screening by simple phenolics like sinapate esters.     

Effect of Ultraviolet- B Irradiation on Lipid Peroxidation in Spinach Leaves

Spinach ( Spinacia oleracea Mill. ) cultivar "Huabo No. 1" was grown in an indoor environment and treated with 13.0 kJ' m-1. d-1 of ultraviolet-B (UV-B 280 to 320 nm) to study the effect of UV-B irradiation on flavonoids and lipid peroxidation in spinach leaves. The results showed that enhanced UV-B irradiation decreased the leaf fresh weight and the content of soluble protein and chlorophyll, and induced large accumulation of UV-absorbing flavonoids in the leaves. UV-B irradiation also promoted the production of superoxide radicals (O2-) and malondialdehyde in spinach leaves. However, the ascorbic acid (ASA) level was decreased under UV-B treatment. It was interesting that high peroxidase (POX), superoxide dismutase (SOD) and catalase (CAT) activities in spinach leaves were induced by UV-B irradiation, the former two being more sensitive. It was suggested that UV-B induced the accumulation of O2- resulting in the lipid peroxidation and in mm inhibiting the growth of spinach. However, the increase of UV-absorbing flavonoids and anti-oxidative enzymes induced by high accumulation of 02- could not reverse the process of UV-B damage.