VEGETOS: An International Journal of Plant Research & Biotechnology
(Society For Plant Research)

Research Articles


Volume: 32, Issue: 4, December 2019

Print ISSN : 0970-4078.
Online ISSN : 2229-4473.
Pub Email:
Page Visits: 32

Doi: 10.1007/s42535-019-00071-y
Doi Link:
First Page: 583
Last Page: 592
Published: 05 November, 2019

Nitric oxide improved salt stress tolerance by osmolyte accumulation and activation of antioxidant defense system in seedling of B. juncea (L.) Czern


The present manuscript elucidates the cytoprotective role of nitric oxide and its relation with ROS and antioxidants in minimizing the effect of salinity in B. juncea (Indian mustard) which is an important salt tolerant crop of arid and semi arid region. NO alleviates the adverse effect of salt on growth, biomass, relative water content and photosynthesis. At 50 mM NaCl concentration, rapid accumulation of NO was observed that is 2.02 folds higher in roots respectively while in leaves and roots insignificant changes were observed. At higher concentration of salt stress (70–100 mM NaCl), considerable rise in H2O2 content and membrane damage were documented which restrained by the application of 100 µM SNP. NO mitigates the salt stress by regulating the activities of nitrate reductase and other antioxidants. Reduction in NO content was observed in independent treatment of NaCl and SNP in comparison to combined treatment of SNP + NaCl (100 µM SNP + 50 mM NaCl). The different antioxidant enzymes show disparate behaviour with signalling molecule NO. The activity of catalase, guaiacol peroxidase and nitrate reductase were upregulated whereas ascorbate peroxidase was downregulated with NO. The elevated activities of antioxidant and NO accumulation might be responsible for detoxifying the detrimental effect of salinity in B. juncea. The current investigation emphasizes on the potential role of NO in salt stress tolerance by regulating antioxidative defense metabolism, cellular homeostasis and detoxification mechanism in seedlings of B. juncea.



Nitric oxide, Salt stress, Stress tolerance, Antioxidant, n Brassica juncean


  1. Aebi H (1974) Catalase. In: Verlag B (ed) Methods of enzymatic analysis. Springer, New York, pp 673–684

  2. Ahmad P, Abdel Latef AA, Hashem A, Abd_Allah EF, Gucel S, Tran L-SP (2016) Nitric oxide mitigates salt stress by regulating levels of osmolytes and antioxidant enzymes in chickpea. Front Plant Sci 7:347

  3. 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

  4. Amooaghaie R, Tabatabaei F, Ahadi A (2013) Role of hematin and sodium nitroprusside in regulating B nigra seed germination under nanosilver and silver nitrate stresses. Ecotoxicol Environ Saf 113:259–270

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

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

  7. 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

  8. Chen J, Xiao Q, Wang C, Wang WH, Wu FH, Chen J, He BY, Zhu Z, Ru QM, Zhang LL, Zheng HL (2014) Nitric oxide alleviates oxidative stress caused by salt in leaves of a mangrove species, Aegiceras corniculatum. Aquat Bot 117:41–47

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

  10. 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

  11. di Toppi LS, Gabbrielli R (1999) Responses to cadmium in higher plants. Environ Exp Bot 41:105–130

  12. Fancy NN, Bahlmann AK, Loake GJ (2017) Nitric oxide function in plant abiotic stress. Plant Cell Environ 40:462–472

  13. Fatma M, Masood A, Per TS, Khan NA (2016) Nitric oxide alleviates salt stress inhibited photosynthetic performance by interacting with sulfur assimilation in mustard. Front Plant Sci 7:521

  14. Gupta P, Srivastava S, Seth CS (2017) 24-Epibrassinolide and sodium nitroprusside alleviate the salinity stress in Brassica juncea L. cv. Varuna through cross talk among proline, nitrogen metabolism and abscisic acid. Plant Soil 411:483–498

  15. Hasanuzzaman M, Oku H, Nahar K, Borhannuddin Bhuyan MHM, Mahmud JA, Baluska F, Fujita M (2018) Nitric oxide-induced salt stress tolerance in plants: ROS metabolism, signalling and molecular interactions. Plant Biotechnol Rep.

  16. Jaiswal A, Singh NK, Shekhawat GS, Akhtar M, Jaiswal JP (2010) Proline biosynthesis and accumulation in plant: a review. Int Trans Appl Sci 2(4):651–660

  17. Khan MN, Siddiqui MH, Mohammad F, Naeem M (2012) Interactive role of nitric oxide and calcium chloride in enhancing tolerance to salt stress. Nitric Oxide 27:210–218

  18. Liang W, Ma X, Wan P, Liu L (2018) Plant salt-tolerance mechanism: a review. Biochem Biophys Res Commun 495:286–291

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

  20. 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

  21. 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

  22. 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.

  23. 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.

  24. 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

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

  26. 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

  27. Romero-Puertas MC, Palma JM, Gomez M, Del Rio LA, Sandalio LM (2002) Cadmium causes the oxidative modification of proteins in pea plants. Plant Cell Environ 25:677–686

  28. Sehar Z, Masood A, Khan NA (2019) Nitric oxide reverses glucose-mediated photosynthetic repression in wheat (Triticum aestivum L.) under salt stress. Environ Exp Bot 161:277–289

  29. Sharma P, Sardana V, Banga SS (2013) Salt tolerance of Indian mustard (Brassica juncea) at germination and early seedling growth. Environ Exp Biol 11:39–46

  30. 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

  31. Su J, Zhang Y, Nie Y, Cheng D, Wang R, Hu H, Chen J, Zhang J, Du Y, Shen W (2019) Hydrogen-induced osmotic tolerance is associated with nitric oxide-mediated proline accumulation and reestablishment of redox balance in alfalfa seedlings. Environ Exp Bot 147:249–260

  32. 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

  33. 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

  34. Verma K, Dixit S, Shekhawat GS, Alam A (2015) Antioxidant activity of heme oxygenase 1 in Brassica juncea (L) Czern (Indian mustard) under salt stress. Turk J Biol 39:540–549

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

  36. Yadu S, Dewangan TL, Chandrakar V, Keshavkant S (2017) Imperative roles of salicylic acid and nitric oxide in improving salinity tolerance in Pisum sativum L. Physiol Mol Biol Plants 23:43–58

  37. Ahmad P, Prasad MNV (2011) Abiotic stress responses in plants: metabolism, productivity and sustainability. Springer, New York

  38. 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 Int J Biol Sci 8(1):48

  39. 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

  40. 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


Acknowledgements :

Author gratefully acknowledges the University Grant Commission, New Delhi for providing financial assistance in the form of Centre for Advanced Study.

Author Information:

Khushboo Khator
Department of Botany, Jai Narain Vyas University, Jodhpur, India

Gyan Singh Shekhawat
Department of Botany, Jai Narain Vyas University, Jodhpur, India

Pdf Download