Skip to main content
SpringerLink
Log in

Cd- and Cu-induced phytotoxicity on 2–3 leaf stage of Cyamopsis tetragonoloba and its regulation by nitrate reductase and ROS quenching enzyme

  • Original Article
  • Published:
Acta Physiologiae Plantarum Aims and scope Submit manuscript

Abstract

The present study focused on Cd- and Cu-induced nitrate reductase activity and its relation to ROS scavenging enzyme in C. tetragonoloba. Hydroponically adapted seedling treated with different metal concentration (Cd and Cu) in the series from 10 to 100 µM. Seedlings were harvested after 96 h of metal treatment and analyze cellular homeostasis and metal tolerance mechanism by determining growth, stress and enzymatic parameters. The nitrate reductase activity was observed to be highest at 70 µM Cd and 50 µM Cu in cyamopsis roots, i.e., 2.31 and 2.59 times higher than control. Exogenous supplementation of SNP (100 µM) enhanced the NR activity in roots of C. tetragonoloba. Polymorphism between different metal treated leaves and roots were also observed. Higher polymorphism was observed in metal-treated roots in comparison to metal-treated leaves. The activity of other antioxidants except catalase (ascorbate peroxidase, guaiacol peroxidase and nitrate reductase was also recorded to be highest at 70 µM Cd and 50 µM Cu. Increased activity of Nitrate reductase and other antioxidants were directly associated with reduction in MDA and H2O2 level at 70 µM Cd and 50 µM Cu. The present investigation is highlighted the defensive role of Nitrate reductase in mitigating Cd and Cu induced toxicity by regulation of antioxidants and ROS metabolism. The study is important as this is the first report exemplifying the protective role of Nitrate reductase in detoxifying ROS in C. tetragonoloba under Cd and Cu stress.

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

Access this article

Log in via an institution

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

CAT:

Catalase

APX:

Ascorbate peroxidase

GPX:

Guaiacol peroxidase

Cd:

Cadmium

Cu:

Copper

H2O2 :

Hydrogen peroxide

MDA:

Malonaldehyde

O2− :

Superoxide radical

ROS:

Reactive oxygen species

NO:

Nitric oxide

SNP:

Sodium nitroprusside

NR:

Nitrate reductase

BCF:

Bioconcentration factor

TF:

Translocation factor

References

  • Aebi H (1974) Catalase, methods of enzymatic analysis. In: Verlag B (ed) Chemie. Academic Press, New York, pp 673–684

    Google Scholar 

  • Ahmad P, Ahanger MA, Alyemeni MN, Wijaya L, Alam P (2018) Exogenous application of nitric oxide modulates osmolyte metabolism, antioxidants, enzymes of ascorbate-glutathione cycle and promotes growth under cadmium stress in tomato. Protoplasma 255:79–93

    CAS  PubMed  Google Scholar 

  • Alexieva V, Sergiev I, Mapelli S, Karanov E (2001) The effect of drought and ultraviolet radiation on growth and stress markers in pea and wheat. Plant Cell Environ 24:1337–1344

    CAS  Google Scholar 

  • Arnon DI (1949) Copper enzymes in isolated chloroplasts: polyphenol oxidases in Beta vulgaris. Plant Physiol 24:1–15

    CAS  PubMed  PubMed Central  Google Scholar 

  • Bates LS, Waldren RP, Tear ID (1973) Rapid determination of free proline for water stress studies. Plant Soil 39:205–207

    CAS  Google Scholar 

  • Bates SS, Tessier A, Campbell PGC, Buffle J (1982) Zinc adsorption and transport by Chlamydomonas variabilis and Scenedesmus subspicatus (Chlorophyceae) grown in semicontinuous culture. J Phycol 18:521–529

    CAS  Google Scholar 

  • Cantu T, Vieira CE, Piffer RD, Luiz GC, de Souza SGH (2016) Transcriptional modulation of genes encoding nitrate reductase in maize (Zea mays) grown under aluminum toxicity. Afri J Biotechnol 15(43):2465–2473

    CAS  Google Scholar 

  • Chen GX, Asada K (1989) Ascorbate peroxidase in tea leaves: occurrence of two isozymes and the differences in their enzymatic and Molecular properties. Plant Cell Physiol 30:987–998

    CAS  Google Scholar 

  • DalCorso G, Manara A, Furini A (2013) An overview of heavy metal challenge in plants: from roots to shoots. Metallomics 5:1117–1132

    CAS  PubMed  Google Scholar 

  • Datta R, Sharma R (1999) Temporal and spatial regulation of nitrate reductase and nitrite reductase in greening maize leaves. Plant Sci 144:77–83

    CAS  Google Scholar 

  • De Vos CHR, Schat H, Vooijs R, Ernst WHO (1989) Copper-induced damage to the permeability barrier in roots of silene cuiubalus. J Plant Physiol 135:164–179

    Google Scholar 

  • Diwan H, Ahmad A, Iqbal M (2012) Cr-induced alterations in photosynthesis and associated attributes in Indian mustard. J Environ Biol 33:239–243

    CAS  PubMed  Google Scholar 

  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12(1):13

    Google Scholar 

  • Gupta KJ, Fernie AR, Kaiser WM, van Dongen JT (2011) On the origins of nitric oxide. Trends Plant Sci 16:160–168

    CAS  PubMed  Google Scholar 

  • Hou LL, Liu F, Zang X, Zhang X, He B, Ding Y, Song X, Xiao D, Wang H (2017) Cloning and transcription analysis of the nitrate reductase gene from Haematococcus pluvialis. Biotechnol Lett 39:589–597

    CAS  PubMed  Google Scholar 

  • Khator K, Shekhawat GS (2018) Regulatory role of thiols and proline in mitigation of Cu induced phytotoxicity in seven day’s old hydroponically acclimatized seedling of Cyamopsis tetragonoloba. Biotech Today Inter J Biol Sci 8(1):48–57

    Google Scholar 

  • Khator K, Shekhawat GS (2019) Nitric oxide improved salt stress tolerance by osmolyte accumulation and activation of antioxidant defense system in seedling of B. juncea (L.) Czern. Vegetos 32:583–592

    Google Scholar 

  • Khator K, Mahawar L, Shekhawat GS (2019) NaCl induced oxidative stress in legume crops of Indian Thar Desert: an insight in the cytoprotective role of HO1, NO and antioxidants. Physiol Mol Biol Plants. https://doi.org/10.1007/s12298-019-00728-7

    Article  PubMed  PubMed Central  Google Scholar 

  • Kumar S, Joshi UN (2008) Nitrogen metabolism as affected by hexavalent chromium in sorghum (Sorghum bicolor L.). Environ Exp Bot 64:135–144

    CAS  Google Scholar 

  • Lillo C, Meyer C, Lea US, Provan F, Oltedal S (2004) Mechanism and importance of post-translational regulation of nitrate reductase. J Exp Bot 65:1275–1282

    Google Scholar 

  • Liu S, Yanga R, Pana Y, Bo R, Chena Q, Xi L, Xi X, Taoa J, Cheng Q, Ma M (2016) Beneficial behavior of nitric oxide in copper-treated medicinal plants. J Hazard Mater 314(2016):140–154

    CAS  PubMed  Google Scholar 

  • Lowry OH, Rosenberg NJ, Farr AL, Randall RJ (1951) Protein measurement with folin phenol reagent. J Biol Chem 193:265–275

    CAS  PubMed  Google Scholar 

  • Mahawar L, Shekhawat GS (2016) Salt induce oxidative stress and its tolerance mechanism in plant: morphological, biochemical and molecular perspective. Biotech Today 6:80–87

    Google Scholar 

  • Mahawar L, Shekhawat GS (2018) Haem oxygenase: a functionally diverse enzyme of photosynthetic organisms and its role in Phytochrome chromophore biosynthesis, cellular signalling and defence mechanisms. Plant Cell Environ 41:483–500

    CAS  PubMed  Google Scholar 

  • Mahawar L, Shekhawat GS (2019) EsHO 1 mediated mitigation of NaCl induced oxidative stress and correlation between ROS, antioxidants and HO 1 in seedlings of Eruca sativa: underutilized oil yielding crop of arid region. Physiol Mol Biol Plants. https://doi.org/10.1007/s12298-019-00663-7

    Article  PubMed  PubMed Central  Google Scholar 

  • Mahawar L, Khator K, Shekhawat GS (2018a) Role of Proline in mitigating NaCl induced oxidative stress in Eruca sativa Miller: an important oil yielding crop of Indian Thar Desert. Vegetos Int J Plant Res Biotechnol. https://doi.org/10.5958/2229-4473.2018.00032.0

    Article  Google Scholar 

  • Mahawar L, Kumar R, Shekhawat GS (2018b) Evaluation of hemeoxygenase 1 (HO 1) in Cd and Ni induced cytotoxicity and crosstalk with ROS quenching enzyme in two to four leaf stage seedling of Vigna radiata. Protoplasma 255:527–545

    CAS  PubMed  Google Scholar 

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

    CAS  Google Scholar 

  • Nabi RBS, Tayade R, Hussain A, Kulkarni KP, Imran QM, Mun BG, Yun BW (2019) Nitric oxide regulates plant responses to drought, salinity, and heavy metal stress. Environ Exp Bot 161:120–133

    CAS  Google Scholar 

  • Nahar K, Rahman M, Hasanuzzaman M, Mahabub Alam M, Rahman A, Suzuki T, Fujita M (2016) Physiological and biochemical mechanisms of spermine-induced cadmium stress tolerance in mung bean (Vigna radiata L.) seedlings. Environ Sci Pollut Res 23:21206–21218

    CAS  Google Scholar 

  • Nanda R, Agrawal V (2016) Elucidation of zinc and copper induced oxidative stress, DNA damage and activation of defence system during seed germination in Cassia angustifolia Vahl. Environ Exp Bot 125:31–41

    CAS  Google Scholar 

  • Olguín EJ, Sánchez-Galván G (2012) Heavy metal removal in phytofiltration and phycoremediation: the need to differentiate between bioadsorption and bioaccumulation. New Biotechnol 30:3–8

    Google Scholar 

  • Pathak R, Singh M, Henry A (2009) Genetic divergence in cluster bean (Cyamopsis tetragonoloba) for seed yield and gum content under rainfed conditions. Indian J Agric Sci 79:559–561

    Google Scholar 

  • Putter J (1974) Peroxidase. In: Bergemeyer HU (ed) Methods of enzymatic analysis. Academic Press, London, pp 685–690

    Google Scholar 

  • Rai V, Vajpayee P, Singh SN, Mehrotra S (2004) Effect of chromium accumulation on photosynthetic pigments, oxidative stress defense system, nitrate reduction, proline level and eugenol content of Ocimum tenuiflorum L. Plant Sci 167:1159–1169

    CAS  Google Scholar 

  • Romero-Puertas M, Palma JM, Gomez M, Dong-Hee L, Sandalio LM (2002) Cadmium causes the oxidative modification of proteins in pea plants. Plant Cell Environ 25:677–686

    CAS  Google Scholar 

  • Samma MK, Zhou H, Cui W, Zhu K, Zhang J, Shen W (2017) Methane alleviates copper-induced seed germination inhibition and oxidative stress in Medicago sativa. Biometals 30:97–111

    CAS  PubMed  Google Scholar 

  • Sandalio LM, Dalurzo HC, Gomez M, Romero-Puertas MC, Del Rio LA (2001) Cadmium-induced changes in the growth and oxidative metabolism of pea plants. J Exp Bot 52:2115–2126

    CAS  PubMed  Google Scholar 

  • Sangwan P, Kumar V, Joshi UN (2014) Effect of chromium (VI) toxicity on enzymes of nitrogen metabolism in clusterbean (Cyamopsis tetragonoloba L.). Enzyme Res 2014:1–9

    Google Scholar 

  • Sanita di Toppi L, Gabbrielli R (1999) Response to cadmium in higher plants. Environ Exp Bot 41:105–130

    Google Scholar 

  • Saxena I, Shekhawat GS (2013) Nitric oxide (NO) in alleviation of heavy metal induced phytotoxicity and its role in protein nitration. Nitric Oxide 32:13–20

    CAS  PubMed  Google Scholar 

  • Schützendübel A, Schwanz P, Teichmann T, Gross K, Langenfeld-Heyser R, Godbold DL, Polle A (2001) Cadmium-induced changes in antioxidative systems, hydrogen peroxide content, and differentiation in scots pine roots. Plant Physiol 127(3):887–898

    PubMed  PubMed Central  Google Scholar 

  • Sharma SS, Dietz KJ (2009) The relationship between metal toxicity and cellular redox imbalance. Trends Plant Sci 14:43–50

    CAS  PubMed  Google Scholar 

  • Shekhawat GS, Prasad A, Verma K, Sharma A (2008) Changes in growth, lipid peroxidation and antioxidant system in seedlings of Brassica juncea (L.) czern. Biochem Cell Arch 8:145–149

    CAS  Google Scholar 

  • Shekhawat GS, Kumar D, Verma K, Singh K, Jana S, Teotia P, Yadav S (2009) Copper as a biometal: an insight on transport, toxicity and tolerance in photosynthetic organisms. In: Blanc G, Moreau D (eds) Biometals: molecular structures, binding properties and applications. Nova Science Publishers, New York (ISBN: 9781608768523)

    Google Scholar 

  • Shekhawat GS, Verma K, Jana S, Singh K, Teotia P, Prasad A (2010) In vitro biochemical evaluation of cadmium tolerance mechanism in callus and seedlings of Brassica juncea. Protoplasma 239:31–38

    CAS  PubMed  Google Scholar 

  • Shekhawat GS, Parihar S, Mahawar L, Khator K, Bulchandani N (2019) Bilin metabolism in plants: Structure, function and Hemeoxygenase regulation of Bilin biosynthesis. eLS 2001:1–13

    Google Scholar 

  • Singh S, Parihar P, Singh R, Singh VP, Prasad SM (2016) Heavy metal tolerance in plants: role of transcriptomics, proteomics, metabolomics and ionomics. Front Plant Sci 6:1143

    PubMed  PubMed Central  Google Scholar 

  • Soda S, Hamada T, Yamaoka Y, Ike M, Nakazato H, Saeki Y, Kasamatsu T, Sakurai Y (2012) Constructed wetlands for advanced treatment of wastewater with a complex matrix from a metal-processing plant: bioconcentration and translocation factors of various metals in Acorus gramineus and Cyperus alternifolius. Ecol Eng 39:63–70

    Google Scholar 

  • Sun C, Lu L, Liu L, Liu W, Yu Y, Liu X, Hu Y, Jin C, Lin X (2014) Nitrate reductase-mediated early nitric oxide burst alleviates oxidative damage induced by aluminum through enhancement of antioxidant defenses in roots of wheat (Triticum aestivum). New Phytol 201:1240–1250

    CAS  PubMed  Google Scholar 

  • Verma K, Shekhawat GS, Sharma A, Mehta SK, Sharma V (2008) Cadmium induced oxidative stress and changes in soluble and ionically bound cell wall peroxidase activities in roots of seedling and 3–4 leaf stage plants of Brassica juncea (L.) Czern. Plant Cell Rep 27:1261–1269

    CAS  PubMed  Google Scholar 

  • Verma K, Mehta SK, Shekhawat GS (2013) Nitric oxide (NO) counteracts cadmium induced cytotoxic processes mediated by reactive oxygen species (ROS) in Brassica juncea: cross-talk between ROS, NO and antioxidant responses. Biometals 26:255–269

    CAS  PubMed  Google Scholar 

  • Wilkins DA (1978) The measurement of tolerance to edaphic factors by means of root growth. New Phytol 80:623–633

    CAS  Google Scholar 

  • Yruela I (2009) Copper in plants: acquisition, transport and interactions. Func Plant Biol 36:409–430

    CAS  Google Scholar 

  • Zhang M, Dong JF, Jin HH, Sun LN, Xu MJ (2011) Ultraviolet-B-induced flavonoid accumulation in Betula pendula leaves is dependent up on nitrate reductase-mediated nitric oxide signaling. Tree Physiol 31:798–807

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

Author would be grateful to acknowledge University Grant Commission, New Delhi for providing financial assistance in the form of Centre for advanced study program.

Author information

Authors and Affiliations

  1. Department of Botany, Jai Narain Vyas University, Jodhpur, Rajasthan, 342005, India

    Khushboo Khator & G. S. Shekhawat

Authors
  1. Khushboo Khator
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. G. S. Shekhawat
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to G. S. Shekhawat.

Additional information

Communicated by S. Esposito.

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Khator, K., Shekhawat, G.S. Cd- and Cu-induced phytotoxicity on 2–3 leaf stage of Cyamopsis tetragonoloba and its regulation by nitrate reductase and ROS quenching enzyme. Acta Physiol Plant 42, 120 (2020). https://doi.org/10.1007/s11738-020-03105-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11738-020-03105-0

Keywords

Search

Navigation