胞外H2O2及NADPH氧化酶参与了铜胁迫对植物细胞死亡的诱导

doi:10.7525/j.issn.1673-5102.2015.05.012

摘要/Abstract

摘要： 以烟草悬浮细胞BY-2(Nicotiana tabacum L.cv.Bright Yellow-2)为材料,探讨了在铜离子胁迫下植物细胞死亡发生过程中胞外H₂O₂及NADPH氧化酶所扮演的角色。实验结果表明,随着外源CuCl₂浓度的上升(从0~700 μmol·L^-1)，细胞死亡水平不断上升，且胞外H₂O₂的水平也不断增加。在300 μmol·L^-1的CuCl₂诱导细胞死亡的过程中,加入H₂O₂清除剂N-N-二甲基硫脲(DMTU)降低了胞外CuCl₂胁迫下H₂O₂含量增加的同时也降低了细胞死亡水平的上升,这一观察表明了铜离子胁迫所导致的细胞死亡的发生和胞外H₂O₂的增加有关。进一步的研究表明,300 μmol·L^-1 CuCl₂的胁迫导致了NADPH氧化酶活性的显著性上升,而加入NADPH氧化酶的抑制剂（二亚苯基碘，DPI,)则降低了CuCl₂胁迫所导致的细胞死亡和胞外H₂O₂含量的上升。上述结果表明，胞外H₂O₂和NADPH氧化酶参与了CuCl₂对植物细胞死亡的诱导作用。

关键词: 细胞死亡, 铜离子, 胞外H₂O₂, NADPH氧化酶

Abstract: By using the tobacco (Nicotiana tabacum cv. Bright Yellow-2) cell suspension culture, we studied the roles of extracellular H₂O₂ and NADPH oxidase in the copper-induced cell death. With the increase of exogenous CuCl₂ from 0 to 700 μmol·L^-1, the level of cell death were increased followed by the increase of extracellular H₂O₂. In the cell subjected to 300 μmol·L^-1 CuCl₂, the addition of exogenous dimethyl thiourea (DMTU, a scavenger of H₂O₂) alleviated the increases of both the cell death level and the production of extracellular H₂O₂, which were induced by CuCl₂(300 μmol·L^-1). The copperinduced increase in the cell death was associated with the increase of extracellular H₂O₂. The CuCl₂(300 μmol·L^-1) stress induced an increase of the activity of NADPH oxidase. The addition of diphenylene iodonium (DPI, an inhibitor of NADPH oxidase) alleviated the increases of both the cell death level and the production of extracellular H₂O₂, which were induced by CuCl₂ (300 μmol·L^-1). Our results indicate that extracellular H₂O₂ and NADPH oxidase are involved in the copper-induced cell death.

Key words: cell death, copper, extracellular H₂O₂, NADPH oxidase

中图分类号:

冯汉青；白晶月；管冬冬；贾凌云；孙坤. 胞外H₂O₂及NADPH氧化酶参与了铜胁迫对植物细胞死亡的诱导[J]. 植物研究, 2015, 35(5): 710-715.

FENG Han-Qing；BAI Jing-Yue；GUAN Dong-Dong；JIA Ling-Yun；SUN Kun. Extracellular H₂O₂ and NADPH Oxidase are Involved in the Copper-Induced Cell Death[J]. Bulletin of Botanical Research, 2015, 35(5): 710-715.

参考文献

1.Henriques F S,Fernandes J C.Metal uptake and distribution in rush(Juncus conglomeratus L.) plants growing in pyrites mine tailings at Lousal,Portugal［J］.Science of the Total Environment,1991,102:253-260.

2.Marschner H.Mineral Nutrition of Higher Plants(Second Edition)［M］.Salt Lake City:Academic Press,1995.

3.Lequeux H,Hermans C,Lutts S,et al.Response to copper excess in Arabidopsis thaliana:Impact on the root system architecture,hormone distribution,lignin accumulation and mineral profile［J］.Plant Physiology and Biochenistry,2010,48:673-682.

4.Raeymaekers T,Potters G,Asard H,et al.Copper-mediated oxidative burst in Nicotiana tabacum L.cv.Bright Yellow 2 cell suspension cultures［J］.Protoplasma,2003,221:93-100.

5.Bartosz M,Kedziora J,Bartosz G.Antioxidant and Prooxidant Properties of Captopril and Enalapril［J］.Free Radical Biology and Medicine,1997,23:729-735.

6.刘登义,王友保,张徐祥,等.污灌对小麦幼苗生长及活性氧代谢的影响［J］.应用生态学报,2002,13(2):179-182.

7.储玲,刘登义,王友保,等.铜污染对三叶草幼苗生长及活性氧代谢影响研究［J］.应用生态学报,2004,1(15):120-122.

8.Chamseddine M,Wided B A,Guy H,et al.Cadmium and copper induction of oxidative stress and antioxidative response in tomato(Solanum lycopersicon) leaves［J］.Plant Growth Regulation,2009,57:89-99.

9.Rhoads D M,Umbach A L,Subbaiah C C,et al.Mitochondrial Reactive Oxygen Species.Contribution to Oxidative Stress and Interorganellar Signaling［J］.Plant Physiology,2006,141:357-366.

10.Tsanko S G,Jacques H.Hydrogen peroxide as a signal controlling plant programmed cell death［J］.Journal of Cell Biology,2005,168:17-20.

11.Sagi M,Fluhr R.Production of Reactive Oxygen Species by Plant NADPH Oxidases［J］.Plant Physiology,2006,141:336-340.

12.Zhang H X,Zhang F Q,Xia Y,et al.Excess copper induces production of hydrogen peroxide in the leaf of Elsholtzia haichowensis through apoplastic and symplastic CuZnsuperoxide dismutase［J］.Journal of Hazardous Materials,2010,178:834-843.

13.Kadono T,Yamaguchi Y,Furuichi T,et al.Ozone-Induced Cell Death Mediated with Oxidative and Calcium Signaling Pathways in Tobacco Bel-W3 and Bel-B Cell Suspension Cultures［J］.Plant Signaling & Behavior,2006,1:312-322.

14.Murashige T,Skoog F.A revised medium for rapid growth and bio assays with tobacco tissue cultures［J］.Physiol Plant,1962,15:473-497.

15.Lin C,Yu Y,Kadono T,et al.Action of aluminum,novel TPC1-type channel inhibitor,against salicylate-induced and cold-shock-induced calcium influx in tobacco BY-2 cells［J］.Biochemical and Biophysical Research Communications,2005,332:823-830.

16.Yang S C.Possible involvement of NADPH oxidase in lanthanide cation-induced superoxide anion generation in BY-2 tobacco cell suspension culture［J］.Journal of Rare Earths,2006,24:243-247.

17.Kawai M,Hirofumi U.Coleoptile senescence in rice(Oryza sativa L.)［M］.Annals of Botany,2000,86:405-414.

18.Patterson B D,MacRae E A,Ferguson I B.Estimation of hydrogen peroxide in plant extracts using titanium(Ⅳ)［M］.Analytical Biochemistry,1984,139:487-492.

19.Kalachova T,Iakovenko O,Kretinin S,et al.Involvement of phospholipase D and NADPH-oxidase in salicylic acid signaling cascade［J］.Plant Physiology and Biochemistry,2013,66C:127-133.

20.王松华,杨志敏,徐朗莱.植物铜素毒害及其抗性机制研究进展［J］.生态环境,2003,12(3):336-341.

21.孙学娟,刘美君,高辉远,等.CuCl₂胁迫对烟草BY-2悬浮细胞死亡的诱导［J］.植物生理学报,2012,48(2):173-180.

22.Meharg A A.Integrated tolerance mechanisms:constitutive and adaptive plant responses to elevated metal concentrations in the environment［J］.Plant Cell and Environment,1994,17:989-993.

23.Achary V M,Panda B B.Aluminium-induced DNA damage and adaptive response to genotoxic stress in plant cells are mediated through reactive oxygen intermediates［J］.Mutagenesis,2010,25:201-209.

24.Drazkiewicz M,Skórzynska-Polit E,Krupa Z.Copper-induced oxidative stress and antioxidant defence in Arabidopsis thaliana［J］.Bio Metals,2014,17:379-387.

[1]	王庆文, 白晶月, 石岱龙, 贾凌云, 冯汉青. 寡霉素处理下细胞外ATP和细胞内ATP的关系及其对细胞死亡调节作用的研究[J]. 植物研究, 2016, 36(6): 925-930.
[2]	汪矛, 崔跃华, 孙克莲. 杜仲胚乳衰退过程中程序性细胞死亡的研究[J]. 植物研究, 1999, 19(4): 401-406.

胞外H₂O₂及NADPH氧化酶参与了铜胁迫对植物细胞死亡的诱导

Extracellular H₂O₂ and NADPH Oxidase are Involved in the Copper-Induced Cell Death

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 2

编辑推荐

Metrics

本文评价