Skip to main content
Log in

Modulation of copper toxicity-induced oxidative damage by nitric oxide supply in the adventitious roots of Panax ginseng

  • Biotic and Abiotic Stress
  • Published:
Plant Cell Reports Aims and scope Submit manuscript

Abstract

Nitric oxide (NO) is a highly reactive, membrane-permeable free radical, which has recently emerged as an important signalling molecule and antioxidant. Here we investigated the protective effect of NO against the toxicity caused by excess CuSO4 (50 μM) in the adventitious roots of mountain ginseng. It was found that NO donor, sodium nitroprusside (SNP), was effective in reducing Cu-induced toxicity in the mountain ginseng adventitious roots. Protective effect of SNP, as indicated by extent of lipid peroxidation, was reversed by incorporation of 2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide (CPTIO), a NO scavenger, in the medium suggesting that the protective effect of SNP is attributable to NO released, which was revealed from in situ confocal laser scanning microscopic localization of NO in the adventitious roots of mountain ginseng. Results obtained in the present study suggest that reduction of excess Cu-induced toxicity by SNP is most likely mediated through the modulation in the activities of antioxidant enzymes involved in H2O2 detoxification (catalase, peroxidase, ascorbate peroxidase) and in the maintenance of cellular redox couples (glutathione reductase), and contents of molecular antioxidants (particularly non-protein thiol, ascorbate and its redox status). Exogenous NO supply also improved the activity of superoxide dismutase, an enzyme responsible for O2 ·− dismutation, and NADPH oxidase, an enzyme responsible for O2 ·− generation, in excess Cu supplied adventitious roots of mountain ginseng.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

APX:

Ascorbate peroxidase

CAT:

Catalase

CPTIO:

2-(4-carboxy-2-phenyl)-4,4,5,5-tetramethyl-imidazoline-1-oxyl-3-oxide

EDTA:

Ethylenediamine tetraacetic acid

GR:

Glutathione reductase

MDA:

Malondialdehyde

POD:

Peroxidase

ROS:

Reactive oxygen species

SNP:

Sodium nitroprusside

SOD:

Superoxide dismutase

NO:

Nitric oxide

TCA:

Trichloroacetic acid

References

  • Ali MB, Hahn EJ, Paek KY (2006) Copper-induced changes in the growth, oxidative metabolism and saponin production in suspension culture roots of Panax ginseng in bioreactors. Plant Cell Rep. 25:1122–1132

    Article  PubMed  CAS  Google Scholar 

  • Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol 55:373–379

    Article  PubMed  CAS  Google Scholar 

  • Arasimowicz M, Floryszak-Wieczorek J (2007). Nitric oxide as a bioactive signalling molecule in plant stress responses. Plant Sci 172:876–887

    Article  CAS  Google Scholar 

  • Baker CJ, Mock NM (1994) An improved method for monitoring cell death in cell suspension and leaf disc assays using Evans blue. Plant Cell Tiss Org Cult 39:7–12

    Article  Google Scholar 

  • Beauchamp C, Fridovich I (1971) Superoxide dismutase: improved assays and an assay applicable to acrylamide gels. Anal Biochem 44:276–287

    Article  PubMed  CAS  Google Scholar 

  • Beligni MV, Lamattina L (1999) Nitric oxide protects against cellular damage produced by methylviologen herbicides in potato plants. Nitric Oxide 3:199–208

    Article  PubMed  CAS  Google Scholar 

  • Beligni MV, Fath A, Bethke PC, Lamattina L, Jones RL (2002) Nitric oxide acts as an antioxidant and delays programmed cell death in barley aleurone layers. Plant Physiol 129:1642–1650

    Article  PubMed  CAS  Google Scholar 

  • Bisht SS, Sharma A, Chaturvedi K (1989) Certain metabolic lesions of chromium toxicity in radish. Indian J Agric Biochem 2:109–115

    CAS  Google Scholar 

  • Boveris AD, Galatro A, Puntarulo S. (2000) Effect of nitric oxide and plant antioxidants on microsomal content of lipid radicals. Biol Res 33:159–165

    Article  PubMed  CAS  Google Scholar 

  • Brennan T, Frenkel C. (1977) Involvement of hydrogen peroxide in regulation of senescence in pear. Plant Physiol 59:411–416

    Article  PubMed  CAS  Google Scholar 

  • Carimi F, Zottini M, Costa A, Cattelan I, Michele RD, Terzi M, Schiavo FL (2005) NO signalling in cytokinin-induced programmed cell death. Plant Cell Environ 28:1171–1178

    Article  CAS  Google Scholar 

  • Corpas FJ, Barroso JB, Carreras A, Quiros M, Leon AM, Romero-Puertas MC, Esteban FJ, Valderrama R, Palma JM, Sandalio LM, Gomez M, del Rio LA (2004) Cellular and subcellular localization of endogenous nitric oxide in young and senescent pea plants. Plant Physiol 136:2722–2733

    Article  PubMed  CAS  Google Scholar 

  • Gapper C, Dolan L (2006) Control of plant development by reactive oxygen species. Plant Physiol 141:341–345

    Article  PubMed  CAS  Google Scholar 

  • Halliwell B. (2006) Reactive species and antioxidants. redox biology is a fundamental theme of aerobic life. Plant Physiol 141:312–322

    Article  PubMed  CAS  Google Scholar 

  • Halliwell B, Gutteridge JMC (1984) Oxygen toxicity, oxygen radicals, transition metals and disease. Biochem J 219:1–14

    PubMed  CAS  Google Scholar 

  • Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplast, I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:180–198

    Google Scholar 

  • Hsu YT, Kao CH (2004) Cadmium toxicity is reduced by nitric oxide in rice leaves. Plant Growth Regul 42:227–38

    Article  CAS  Google Scholar 

  • Hung KT, Kao CH (2003) Nitric oxide counteracts the senescence of rice leaves induced by abscisic acid. J Plant Physiol 160:871–879

    Article  PubMed  CAS  Google Scholar 

  • Hung KT, Kao CH (2004) Nitric oxide acts as an antioxidant and delays methyl jasmonate-induced senescence of rice leaves. J Plant Physiol 161:43–52

    Article  PubMed  CAS  Google Scholar 

  • Hung KT, Chang CJ, Kao CH (2002). Paraquat toxicity is reduced by nitric oxide in rice leaves. J Plant Physiol 159:159–166

    Article  CAS  Google Scholar 

  • Jablonski PP, Anderson JW (1978) Light-dependent reduction of oxidised glutathione by ruptured chloroplasts. Plant Physiol 61:221–225

    PubMed  CAS  Google Scholar 

  • Kojima H, Nakatsubo N, Kikuchi K, Kawahara S, Kirino Y, Nagoshi H, Hirata Y, Nagano T. (1998) Detection and imaging of nitric oxide with novel fluorescent indicators: diaminofluoresceins. Anal Chem 70:2446–2453

    Article  PubMed  CAS  Google Scholar 

  • Lamattina L, Garcia-Mata C, Graziano M, Pagnussat G (2003) Nitric oxide: the versatility of an extensive signal molecule. Annu Rev Plant Biol 54:109–136

    Article  PubMed  CAS  Google Scholar 

  • Laspina NV, Groppa MD, Tomaro ML, Benavides MP (2005) Nitric oxide protects sunflower leaves against Cd-induced oxidative stress. Plant Sci 169:323–330

    Article  CAS  Google Scholar 

  • Liszkay A, van der Zalm E, Schopfer P. (2004) Production of reactive oxygen intermediates O2 ·−, H2O2, and OH˙ by maize roots and their role in wall loosening and elongation growth. Plant Physiol 136:3114–3123

    Article  PubMed  CAS  Google Scholar 

  • Maksymiec W (1997) Effect of copper on cellular processes in higher plants. Photosynthetica 34:321–342

    Article  CAS  Google Scholar 

  • Maksymiec W, Krupa Z. (2006) The effects of short-term exposition to Cd, excess Cu ions and jasmonate on oxidative stress appearing in Arabidopsis thaliana. Environ Exp Bot. 57:187–194

    Article  CAS  Google Scholar 

  • Marschner H (1995) Mineral nutrition of higher plants. Academic, London

    Google Scholar 

  • Murashige T., Skoog F. (1962) A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Murgia I, de Pinto MC, Delledonne M, Soave C, Gara LD (2004) Comparative effects of various nitric oxide donors on ferritin regulation, programmed cell death, and cell redox state in plant cells. J Plant Physiol 161:777–783

    Article  PubMed  CAS  Google Scholar 

  • Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate specific peroxidase in spinach chloroplast. Plant Cell Physiol 22:867–880

    CAS  Google Scholar 

  • Orozco-Cardenas M, Ryan CA (2002) Nitric oxide negatively modulates wound signaling in tomato plants. Plant Physiol 130:487–93

    Article  PubMed  CAS  Google Scholar 

  • Pätsikkä E, Kairavuo M, Šeršen F, Aro E-M, Tyystjärvi E. (2002) Excess copper predisposes photosystem II to photoinhibition in vivo by outcompeting iron and causing decrease in leaf chlorophyll. Plant Physiol 129:1359–1367

    Article  PubMed  CAS  Google Scholar 

  • Quartacci MF, Cosi E, Navari-Izzo F (2001) Lipid and NADPH-dependent superoxide production in plasma membrane vesicles from roots of wheat grown under copper deficiency or excess. J Exp Bot 52:77–84

    Article  PubMed  CAS  Google Scholar 

  • Rao MV, Hale BA, Ormrod DP (1995) Amelioration of ozone-induced oxidative damage in wheat plants grown under high carbon dioxide. Plant Physiol 109:421–32

    PubMed  CAS  Google Scholar 

  • Rubbo H, Radi R, Anselmi D, Kirk M, Barnes S, Butler J, Eiserich JP, Freeman BA (2000) Nitric oxide reaction with lipid peroxyl radicals spares alpha-tocopherol during lipid peroxidation. Greater oxidant protection from the pair nitric oxide/alphatocopherol than alpha-tocopherol/ascorbate. J Biol Chem 275:10812–10818

    Article  PubMed  CAS  Google Scholar 

  • Sagi M, Fluhr R (2001) Superoxide production by plant homologues of the gp91(phox) NADPH oxidase: modulation of activity by calcium and by tobacco mosaic virus infection. Plant Physiol 126:1281–1290

    Article  PubMed  CAS  Google Scholar 

  • Sagi M, Fluhr R (2006) Production of reactive oxygen species by plant NADPH oxidases. Plant Physiol 141:336–340

    Article  PubMed  CAS  Google Scholar 

  • Sarath G, Hou G, Baird LM, Mitchell RB (2007) Reactive oxygen species, ABA and nitric oxide interactions on the germination of warm-season C4-grasses. Planta doi 10.1007/s00425-007-0517-z

  • Sharma SS, Dietz KJ, (2006) The significance of amino acids and amino acid-derived molecules in plant responses and adaptation to heavy metal stress J Exp Bot 57:711–726

    Article  PubMed  CAS  Google Scholar 

  • Singh AK, Sharma L, Mallick N (2004) Antioxidative role of nitric oxide on copper toxicity to a chlorophycean alga, Chlorella. Ecotoxicol Environ Safe 59:223–227

    Article  CAS  Google Scholar 

  • Suh Y.-A., Arnold RS, Lassegue B, Shi J, Xu X, Sorescu D, Chung AB, Griendling KK, Lambeth JD (1999) Cell transformation by the superoxide-generating oxidase Mox1. Nature 401:79–82

    Article  PubMed  CAS  Google Scholar 

  • Takahama U, Oniki T (1992) Regulation of peroxidase-dependent oxidation of phenolics in the apoplast of spinach leaves by ascorbate. Plant Cell Physiol 33:279–387

    Google Scholar 

  • Tewari RK, Kumar P, Sharma PN (2006) Antioxidant responses to enhanced generation of superoxide anion radical and hydrogen peroxide in the copper-stressed mulberry plants. Planta 223:1145–1153

    Article  PubMed  CAS  Google Scholar 

  • Wink DA, Mitchell JB (1998) Chemical biology of nitric oxide: insights into regulatory, cytotoxic, and cytoprotective mechanisms of nitric oxide. Free Radic Biol Med 25:434–456

    Article  PubMed  CAS  Google Scholar 

  • Wink DA, Cook JA, Pacelli R, Liebmann J, Krishne MC, Mitchell JB (1995) Nitric oxide (NO) protects against cellular damage by reactive oxygen species. Toxicol Lett 82–83:221–226

    Article  PubMed  Google Scholar 

  • Xiang C, Oliver DJ (1998) Glutathione metabolic genes coordinately respond to heavy metals and jasmonic acid in Arabidopsis. Plant Cell 10:1539–1550

    Article  PubMed  CAS  Google Scholar 

  • Yu KW, Gao WY, Son SH, Paek KY (2000) Improvement of ginsenoside production by jasmonic acid and some other elicitors in hairy root culture of ginseng (Panax ginseng C.A. Meyer). In Vitro Cell Develop Biol Plant 36:424–428

    Article  CAS  Google Scholar 

  • Yu CC, Hung KT, Kao CH (2005) Nitric oxide reduces Cu toxicity and Cu-induced NH +4 accumulation in rice leaves. J Plant Physiol 162:1319–1330

    Article  PubMed  CAS  Google Scholar 

  • Zhou B, Guo Z, Xing J, Huang B (2005) Nitric oxide is involved in abscisic acid-induced antioxidant activities in Stylosanthes guianensis. J Exp Bot 56:3223–3228

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This laboratory work is financially supported by the Ministry of Education and Human Resource Development (MOE), the Ministry of Commerce, Industry and Energy (MOCIE), Ministry of Labor (MOLAB) and the Korea Science and Engineering Foundation (KOSEF) grant funded by Korea government (MOST). Special thanks are given to Mrs. J-S Jeon for helpful technical assistance.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Kee-Yoeup Paek.

Additional information

Communicated by W.T. Kim.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Tewari, R.K., Hahn, EJ. & Paek, KY. Modulation of copper toxicity-induced oxidative damage by nitric oxide supply in the adventitious roots of Panax ginseng . Plant Cell Rep 27, 171–181 (2008). https://doi.org/10.1007/s00299-007-0423-7

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00299-007-0423-7

Keywords

Navigation