887 resultados para antioxidant


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When tobacco BY-2 cells were treated with 60 mu g/mL MC-RR for 5 d, time-dependent effects of MC-RR on the cells were observed. Morphological changes such as abnormal elongation, evident chromatin condensation and margination, fragmentation of nucleus and formation of apoptotic-like bodies suggest that 60 mu g/mL MC-RR induced rapid apoptosis in tobacco BY-2 cells. Moreover, there was a significant and rapid increase of ROS level before the loss of mitochondrial membrane potential (Delta Psi(m)) and the onset of cell apoptosis. Ascorbic acid (AsA), a major primary antioxidant, prevented the increase of ROS generation, blocked the decrease in Delta Psi(m) and subsequent cell apoptosis, indicating a critical role of ROS in serving as an important signaling molecule by causing a reduction of Delta Psi(m) and MC-RR-induced tobacco BY-2 cell apoptosis. In addition, a specific mitochondrial permeability transition pores (PTP) inhibitor, cyclosporin A (CsA), significantly blocked the MC-RR-induced ROS formation, loss of Delta Psi(m), as well as cell apoptosis when the cells were MC-RR stressed for 3 d, suggesting that PTP is involved in 60 mu g/mL MC-RR-induced tobacco cell apoptosis signalling process. Thus, we concluded that the mechanism of MC-RR-induced apoptosis signalling pathways in tobacco BY-2 cells involves not only the excess generation of ROS and oxidative stress, but also the opening of PTP inducing loss of mitochondrial membrane potential. (C) 2007 Published by Elsevier Ltd.

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The allelopathic effects of two submerged macrophytes, Najas minor and Potamogeton malaianus, on growth, photosynthesis and antioxidant systems of Scenedesmus obliquus were assessed in coexistence experiments. The growth of S. obliquus was significantly suppressed by the two macrophytes. Moreover, P. malaianus showed the stronger growth inhibition effect on S. obliquus than N. minor. P. malaianus obviously inhibited the photosynthetic rate of S. obliquus, while N. minor had no inhibitory effect. Lipid peroxidation and three antioxidant enzymes activities (superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD)) of S. obliquus were investigated at the end of the co-cultures. The two macrophytes significantly enhanced the malondialdehyde (MDA) content, a product of lipid peroxidation, in S. obliquus. Activities of the three antioxidant enzymes of S. obliquus were simultaneously stimulated in P. malaianus treatment, while no significant variation of POD activity was observed in N. minor treatment. The results indicated that the two macrophytes N. minor and P. malaianus had significant allelopathic effects on S. obliquus. However, the two macrophytes influenced S. obliquus in different ways.

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The objective of this study was to evaluate the sub-lethal toxicity of hexabromocyclododecane (HBCDD) in fish. Adult Chinese rare minnows as in vivo models were exposed to waterborne HBCDD from 1 to 500 mu g/l for 14, 28 and 42 days. Hepatic CYP1A1 (ethoxyresorufin-O-deethylase, EROD) and CYP2B1 (pentaoxyresorufin-O-depentylase, PROD) activities were measured. At the same time, molecular biomarkers of oxidative stress were also assayed in the brain, including reactive oxygen species (ROS), lipid peroxidation products (thiobarbituric acid-reactive substances, TBARS), DNA damage and protein carbonyl, as well as superoxide dismutase (SOD) activity and glutathione (GSH) content. DNA damage was evaluated using the Comet assay on erythrocytes. Besides, the content of HBCDD in whole fish was determined after 42 days exposure. The results show that HBCDD could induce EROD and PROD at 500 mu g/l after 28 days exposure, and at 100 to 500 mu g/l after 42 days exposure (P < 0.05), respectively. ROS formation in fish brain was observed to be increased in both time- and dose-dependent manner due to HBCDD exposure. The significant increases in TBARS and protein carbonyl contents occurred in fish brain after 28 and 42 days exposure (P < 0.05). Significant DNA damage in erythrocytes by Comet assay was also found in the 100-500 mu g/l exposure groups (P < 0.05) after 42 days exposure. Moreover, significant depletion in brain GSH content occurred in all treated groups (P < 0.05) and apparent inhibition in SOD activity in brain was observed in the groups of 10-500 mu g/l concentrations during 42 days exposure. The results demonstrate that increasing duration of HBCDD exposure induced EROD and PROD activities, caused excess ROS formation, finally resulted in oxidative damage to lipids, proteins and DNA and decreased antioxidant capacities in fish. Chemical analysis of HBCDD in whole fish showed accumulation up to 654 mu g/g wet weight. (c) 2007 Elsevier B.V. All rights reserved.

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Physiological and biochemical responses of four fishes with different trophic levels to toxic cyanobacterial blooms were studied in a large net cage in Meiliang Bay, a hypereutrophic region of Lake Taihu. We sampled four fishes: the phytoplanktivorous Hypophthalmichthys molitrix and Aristichthys nobilis, the omnivorous Carassius auratus, and the carnivorous Culter ilishaeformis. Alterations of the antioxidant (GSH) and the major antioxidant enzymes (CAT, SOD, GPx, GST) in livers were monitored monthly, and the ultrastructures of livers were compared between the bloom and post-bloom periods. During the cyanobacterial blooms, the phytoplanktivorous fishes displayed only slight ultrastructural changes in liver, while the carnivorous fish presented the most serious injury as swollen endomembrane system and morphologically altered nuclei in hepatocytes. Biochemically, the phytoplanktivorous fishes possessed higher basal GSH concentrations and better correlations between the major antioxidant enzymes in liver, which might be responsible for their powerful resistance to MCs. This article provided physiological and toxicological evidences for the possible succession of fish communities following occurrence of toxic cyanobacterial blooms and also for the applicability of using phytoplanktivorous fish to counteract toxic cyanobacterial blooms in natural waters. (C) 2007 Elsevier Ltd. All rights reserved.

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Polybrominated diphenyl ethers (PBDEs) are an important class of halogenated organic brominated flame retardants. Because of their presence in abiotic and biotic environments widely and their structural similarity to polychlorinated biphenyls (PCBs), concern has been raised on their possible adverse health effects to humans. This study was designed to determine the anti-proliferative, apoptotic properties of decabrominated diphenyl ether (PBDE-209), using a human hepatoma Hep G2 line as a model system. Hep G2 cells were cultured in the presence of PBDE-209 at various concentrations (1.0-100.0 mu mol/L) for 72 h and the percentage of cell viability was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. The results showed that PBDE-209 inhibited the cells viability in time and concentration-dependent characteristics at concentrations (10.0-100.0 mu mol/L). We found that anti-proliferative effect of PBDE-209 was associated with apoptosis on Hep G2 cells by determinations of morphological changes, cell cycle and apoptosis. Mechanism study showed that PBDE-209 could increase the generation of intracellular reactive oxygen species (ROS) concentration-dependently. Antioxidant N-acetylcyteine partially inhibited the increase of ROS. The mechanism for its hepatoma-inhibitory effects was the induction of cellular apoptosis through ROS generation. In addition, activity of lactate dehydrogenase (LDH) release increased when the cells incubated with PBDE-209 at various concentrations and times. These results suggested that PBDE-209 had the toxicity activity of anti-proliferation and induction of apoptosis in tumor cells in vitro. (c) 2007 Elsevier Ireland Ltd. All rights reserved.

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Allelopathic effects of the submerged macrophyte Potamogeton malaianus on Scenedesmus obliquus were assessed using a twophase approach under controlled laboratory conditions. In the co- culture experiment ( phase I), the growth of S. obliquus at two different initial cell densities was significantly inhibited by P. malaianus. Moreover, the growth inhibition was dependent on the biomass density of P. malaianus. Antioxidant enzymes ( SOD, CAT and POD), MDA, APA, total soluble protein, protein electrophoretic pattern and morphology of S. obliquus were determined after the coculture experiment was terminated. The activities of SOD, CAT, POD and APA at the low initial cell density were stimulated, the contents of MDA and total soluble protein were increased, and some special protein bands disappeared in P. malaianus treatments. The macrophyte had no effect on the activities of SOD and APA at the high initial cell density, but significantly influenced other physiological parameters of S. obliquus with the increase of biomass density. The morphology of S. obliquus showed no difference in the macrophyte treatments and the controls, and the cultures were dominated by 4- celled coenobia. The results indicated P. malaianus had significant allelopathic effects on the growth and physiological processes of S. obliquus. Moreover, the allelopathic effects depended on initial algal cell density, biomass density of the macrophyte, and their interaction. In the experiment of P. malaianus culture filtrates ( phase II), filtrates from combined culture of plant and S. obliquus at the low initial cell density exhibited no apparent growth inhibitory effect on S. obliquus. The result showed that initial addition of growth- inhibiting plant filtrates had no allelopathic effect on S. obliquus. We concluded that the allelopathic effects on S. obliquus were found in the presence of P. malaianus, but not in P. malaianus filtrates. However, the absence of allelopathic effect on S. obliquus might be due to the very low concentrations of allelochemicals in the filtrates.

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Many experimental studies have documented the impact of microcystins (MC) on fish based on either intraperitoneal injection, or oral gavaging via the diet, but few experiments were conducted by MC exposure through natural food uptake in lakes. In this study, the phytoplanktivorous silver carp were stocked in a large pen set in Meiliang Bay of Taihu Lake where toxic Microcystis blooms occurred in the warm seasons. Fish samples were collected monthly and MC concentrations in liver and kidney of the fish were determined by LC-MS. The maximum MC concentrations in liver and kidney were present in July when damages in ultrastructures of the liver and kidney were revealed by electron microscope. In comparison with previous studies on common carp, silver carp showed less damage and presence of lysosome proliferation in liver and kidney. Silver carp might eliminate or lessen cell damage caused by MC through lysosome activation. Recovery in the ultrastructures of liver and kidney after Microcystis blooms was companied with a significant decrease or even disappearance of MC. Catalase and glutathione S-transferase in liver and kidney of silver carp during Microcystis blooms were significantly higher than before and after Microcystis blooms. The high glutathione pool in liver and kidney of silver carp suggests their high resistance to MC exposure. The efficient antioxidant defence may be an important mechanism of phytoplanktivorous fish like silver carp to counteract toxic Microcystis blooms. (C) 2007 Published by Elsevier Ltd.

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The objective of this study was to determine the effect of dietary vitamins A, D-3, E, and C on the gonad development, lipid peroxidation, and immune response of yearling rice field eel, Monopterus albus. A 6-wk feeding trial was designed according to an L-16(4(5)) orthogonal design, in which four vitamins, each at four supplementation levels, were arranged. Sixteen diets were mixed with the different vitamin levels and randomly assigned to 16 groups of fish. Increasing dietary vitamin E supplementation level significantly (P <= 0.05) increased the gonadosomatic index and lowered the serum content of malondialdehyde of rice field eel. Increasing dietary vitamin A and C levels also showed similar effect, but the differences were not statistically significant. Serum immunoglobulin M content increased significantly (P <= 0.01) as dietary vitamin C supplementation levels increased. The concentrations of calcium in bones showed significant (P <= 0.05) trend with vitamin D-3 and A supplementation levels, but the bone phosphorus content was not affected by the dietary vitamin levels.

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A copper/zinc superoxide dismutase (Cu/ZnSOD) gene and a manganese superoxide dismutase (MnSOD) gene of the human parasite Clonorchis sinensis have been cloned and their gene products functionally characterized. Genes Cu/ZnSOD and MnSOD encode proteins of 16 kDa and 25.4 kDa, respectively. The deduced amino acid sequences of the two genes contained highly conserved residues required for activity and secondary structure formation of Cu/ZnSOD and MnSOD, respectively, and show up to 73.7% and 75.4% identities with their counterparts in other animals. The genomic DNA sequence analysis of Cu/ZnSOD gene revealed this as an intronless gene. Inhibitor studies with purified recombinant Cu/ ZnSOD and MnSOD, both of which were functionally expressed in Escherichia coli, confirmed that they are copper/zinc and manganese-containing SOD, respectively. Immunoblots showed that both C. sinensis Cu/ZnSOD and MnSOD should be antigenic for humans, and both, especially the C. sinensis MnSOD, exhibit extensive cross-reactions with sera of patients infected by other trematodes or cestodes. RT-PCR and SOD activity staining of parasite lysates indicate that there are no significant differences in mRNA level or SOD activity for both species of SOD, indicating cytosolic Cu/ZnSOD and MnSOD might play a comparatively important role in the C. sinensis antioxidant system.

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Acute biochemical responses of Potamogeton crispus L. to high external ammonium were investigated in an aquarium experiment. Shoots of P. crispus were incubated in aquaria for 24 h or 48 h at five treatments of ammonium-0, 1, 5, 10 and 20 mg/L NH4-N. Soluble sugar content of the shoots declined markedly with increasing ammonium levels, whereas soluble amino acid content increased dramatically. Responses of two antioxidant enzymes as well as soluble protein content fit a lognormal distribution with increasing ammonium levels. High ammonium levels (NH4-N greater than or equal to 5 mg/L) caused significant acute biochemical changes in P. crispus, which potentially could lead to significant biochemical damage.

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探讨盐胁迫对苜蓿幼苗不同器官生长及抗氧化酶系统的影响,为苜蓿植被恢复技术的研发提供理论参考。【方法】以新牧一号(盐忍耐品种)和北极星(盐敏感品种)2个苜蓿品种为材料,以0mmol/L NaCl处理为对照,用200mmol/L NaCl胁迫处理,测定萌发7d苜蓿幼苗芽、根生长及其H2O2、丙二醛(MDA)含量,相对质膜透性、抗氧化保护酶活性及其同功酶的变化。【结果】200mmol/L NaCl胁迫抑制了苜蓿幼苗的生长,并导致芽、根H2O2、MDA含量及相对质膜透性升高,但新牧一号芽、根器官均表现出比北极星较低程度的生长抑制或质膜损伤。盐胁迫后超氧化物歧化酶(SOD)、抗坏血酸过氧化物酶(APX)、过氧化氢酶(CAT)、过氧化物酶(POD)的活性均明显增加。CAT和POD对活性氧的清除具有器官差异性,CAT活性在芽中较强,POD则与之相反。SOD-Ⅱ、APX-Ⅰ、POD-Ⅲ及POD-Ⅳ为芽、根中盐胁迫敏感的同功酶谱带。【结论】苜蓿盐忍耐品种通过更强的抗氧化保护能力,降低了盐胁迫对其幼苗的损伤。

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首次采用高速逆流色谱技术分离了车前子中的两个苯乙醇昔化合物,所用的溶剂系统简单,快速,高效,避免了传统方法分离单体化合物耗时以及与固态载体表面不可逆吸附等缺点,为大量制备分离这两个化合物提供了一种简单有效的方法。利用高效液相色谱一大气压化学电离质谱联用技术,对其化学结构进行推断和确认,提出了两种化合物可能的质谱碎裂规律。采用三种不同的抗氧化活性体外评价体系,首次对车前子80%甲醇提取物以及从中分离得到的两个化合物的抗氧化活性进行了系统研究。研究结果表明:它们都具有良好的抗氧化活性,而且证明车前子80%甲醇提取物的抗氧化活性主要来自这两个化合物的贡献。采用电喷雾质谱技术,对车前草和肉从蓉两种药材中的苯乙醇营化合物的特征质谱行为进行了系统研究,总结了该类化合物特征的质谱裂解规律,建立了这一类化合物的快速鉴定的质谱新方法。首次通过高效液相色谱一电喷雾质谱联用技术,研究了车前草中苯乙醇营化合物,建立了一种快速、简便地鉴别车前草粗提物中苯乙醇昔化合物的新方法。采用高效液相色谱一电喷雾质谱联用技术(蒸发光检测器和电喷雾质谱检测器),对三七,人参和西洋参中的皂营类化合物进行了系统的分析研究,并建立了三种中药粗提物中皂营类化合物快速检测的新方法。

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近年来,随着对作物重茬障碍原因的深入研究,植物的化感作用越来越受到国内外众多学者的重视。花椒(Zanthoxy piperitum.)为芸香科植物,是一种收益早、用途广、价值高的经济树种,是川西干旱河谷地区的重要经济作物,其连作障碍也倍受关注,系统研究花椒化感作用将有助于理解和最终解决花椒连作障碍问题。本文首先通过萃取、层析等方法分离花椒主效化感成分;通过外加不同浓度的花椒叶水浸液研究了对土壤氮素养分循环的影响;研究了花椒叶水浸液对苜蓿生理生化、光合作用、氮素养分吸收的影响,并对外施氮肥对这种化感影响的缓解作用做了研究;研究了花椒化感潜力对全球变化——UV-B增强辐射的响应。主要研究结果如下: 1.用不同极性的有机溶剂对花椒叶水浸液浓缩浸膏萃取、柱层析,结合生物活性检测,分离得到主效化感作用组分的一种化感物质——对甲氧基苯酚。采用该物质纯品进行生物活性检测,证明其具有化感作用。 2.花椒叶水浸液处理土壤30天后,土壤硝态氮、铵态氮、无机氮(硝态氮+铵态氮)与对照相比,随着花椒叶水浸液浓度的增加呈现降低的趋势,其中土壤铵态氮含量显著降低,而硝态氮含量的变化则不显著,无机氮含量也显著降低。土壤脲酶和蛋白酶的活性与无机氮含量的变化趋势相同。随着花椒叶水浸液浓度的增加,氨化细菌数量显著降低,固氮菌的数量变化不显著,硝化细菌和反硝化细菌数量有减少的趋势。60天后,硝态氮含量、铵态氮含量、无机氮随水浸液浓度增加的变化趋势与30天时相似;随着花椒叶水浸液浓度的增加,氨化细菌、固氮菌的数量显著减少,硝化细菌数量、反硝化细菌数量仍呈减少趋势;土壤脲酶、蛋白酶活性与第30天的变化趋势相同。第60天与第30天的结果相比,相同水浸液浓度处理的硝态氮、铵态氮、无机氮均有下降的趋势,但除了25g.L-1水浸液处理的外,其它相同浓度的处理间差异均不显著;除了12.5 g.L的处理外土壤脲酶活性均呈增强的趋势;蛋白酶活性都有不同程度的增加;花椒叶水浸液处理的土壤硝化细菌和反硝化细菌数量呈增加趋势。 3.随着花椒叶水浸液浓度的增加,显著抑制了苜蓿根长、地上地下生物量、叶绿素含量、叶片中可溶性蛋白的含量,净光合速率。苜蓿体内四种抗氧化酶(POD、SOD、CAT、APX) 活性随着水浸液浓度的增加而降低,而丙二醛含量则增加。苜蓿氮初级同化相关酶硝酸还原酶(NR)、谷氨酰合成酶(GS)、谷氨酸脱氢酶(GDH)的活性随着水浸液浓度的增加受到不同程度的影响。总的来说,苜蓿硝酸还原酶、谷氨酰合成酶的活性受到抑制,而谷氨酸脱氢酶活性的变化则比较复杂,根呈先降低后增加的趋势,叶片则无显著变化。外施两种不同浓度的硝酸铵氮肥后,对12.5、25 g.L-1花椒叶水浸液处理的苜蓿化感作用有显著的缓解作用,表现在株高、生物量、光合作用等方面,大多达到与对照(0 g.L-1)未施氮肥无显著差异的水平,而对50 g.L-1水浸液处理的苜蓿幼苗,虽有一定的缓解作用,但这种作用均未达到与对照(0 g.L-1)未施氮肥时无显著差异的水平。 4. UV-B增强辐射处理花椒后,花椒的化感潜力显著增强。花椒叶片内UV-B吸收物质的含量和总酚含量均显著增加。 In recent years, with profound research on the reasons of continuous cropping obstacles, allelopathy received increasing attention to many scholars at home and abroad. Zanthoxy bungeanum as a Rutaceae plant is a high economic value species which gains early and uses widely. Zanthoxylum is an important economic crop in the arid valley of western Sichuan region, and its not even has received much concern for the continuous cropping obstacles. The systematic study of allelopathy of Zanthoxylum will contribute to the understanding and final settlement of this issue. The major allelopathic composition was separated through the extraction, chromatography combined with other methods. The impact on soil nutrient cycling was also studied through the addition of different concentrations of water extracts of Zanthoxylum. Furthermore, the effects of water extracts of Zanthoxylum leaves on alfalfa leaf physiological and biochemical indexes, photosynthesis, soil enzymes and nutrient uptake of nitrogen and the mitigation of allelopathy through using external fertilizer were studied to put forward scientific resolvent for Zanthoxylum continuous cropping obstacles .The response of allelopathic potential of Zanthoxylum to global change - UV-B enhanced radiation was studied . The main findings are as follows: 1. Through extraction with different polar organic solvents on concentrated water extract of Zanthoxylum leaf and then using column chromatography combined with detection of biological activity, one of the main allelopathic components- methoxy-phenol was isolated. The biological activity testing of the pure material of methoxy-phenol proved that it does have allelopathic potential. 2. Thirty days after treating soil with water extract of Zanthoxylum leaf, as compared with the control, the contents of soil nitrate, ammonium, nitrate plus ammonium nitrogen showed a trend of decrease with the increase of the concentration of water extract whereas the content of ammonium nitrogen showed a significant reduction, and the content of nitrate did not change significantly, the content of nitrate plus ammonium nitrogen also showed a significant (P <0.05) redction. The activity of soil urease and protease showed the same trend as the content of nitrate nitrogen plus ammonium nitrogen. With the increase in the concentration of water extract, the number of ammonification bacteria significantly reduced but nitrogen-fixing bacteria did not change significantly and there was a decreasing trend in the number of nitrifying bacteria and denitrifying bacteria. Sixty days after the treatment, with the increase in solution concentration of water extract of Zanthoxylum leaf, the content of nitrate、 ammonium nitrogen, nitrate plus ammonium nitrogen showed a similar change trend to 30 days’; the number of ammonification bacteria, nitrogen-fixing bacteria significantly reduced ; the number of nitrifying bacteria, denitrifying bacteria was still an downward trend; the activity of soil urease and protease showed the same trend as the 30th days’. Compared to the results of the 30th days’, the content of nitrate, ammonium, nitrate plus ammonium nitrogen showed a decrease trend between the treatment of same concentration, but there was no significant difference except the treatment of 25g.L-1 between the same concentration; the activity of soil urease showed enhanced trend except the treatment of 12.5 g.L-1; the activity of protease increased to varying degrees; the number of ammonification bacteria、 nitrifying bacteria and denitrifying bacteria were growing while nitrogen-fixing bacteria reduced.. 3. With the increase of the concentration of water extract of Zanthoxylum leaf, the water extract significantly inhibited the root length, aboveground biomass, content of chlorophyll and soluble protein in leaf and net photosynthetic rate. The activity of four antioxidant enzymes (POD, SOD, CAT, APX) reduced with the increase in concentration of the water extract but the content of MDA increased. The activity of enzymes related to primary nitrogen assimilation such nitrate reductase (NR), glutamyl synthetase (GS), glutamate dehydrogenase (GDH) were subject to different degrees with an increase in the concentration of water extracts. In general, the activity of nitrate reductase, glutamyl synthetase were inhibited, while change in the activity of glutamate dehydrogenase was more complex. The activity of glutamate dehydrogenase in leaf was first reduced and then increase,but did not change significantly in root. After using two external different concentrations of nitrogen fertilizer, there was a significant mitigation in inhibiton in plant height, biomass, photosynthesis, etc. in the treatment of 12.5,25 gL-1 of water extract of Zanthoxylum leaf, and most of these indexes showed no significant difference with the control (0 g.L-1, no external fertilizer was added) .Although there showed a certain degree of ease in the treatment of 50 g.L-1 , there was still a significant difference compared with the control (0 gL-1) in which no external fertilizer was used. 4.The allelopathic potential of Zanthoxylum positively responded to enhanced UV-B significantly. The content of UV-B absorbing compounds and the total phenol also significant increased.