Acta Physiologiae Plantarum

, Volume 35, Issue 4, pp 1039–1050 | Cite as

Changes in growth, lipid peroxidation and some key antioxidant enzymes in chickpea genotypes under salt stress

  • Saiema Rasool
  • Altaf Ahmad
  • T. O. Siddiqi
  • Parvaiz Ahmad
Original Paper


The present study was conducted to evaluate the effect of NaCl on growth and some key antioxidants in chickpea. Eight genotypes of chickpea were grown hydroponically for 15 days and then treated with different concentrations of salt [0 mM (T0), 25 mM (T1), 50 mM (T2), 75 mM (T3), and 100 mM (T4)]. Salinity showed marked changes in growth parameters (fresh and dry weight of root and shoot). The level of lipid peroxidation was measured by estimating malondialdehyde content. Lipid peroxidation increases with the increase in NaCl concentration in all genotypes but salt-tolerant genotypes (SKUA-06 and SKUA-07) were least affected as compared to other genotypes. The chlorophyll content was also affected with elevated levels of NaCl. Increased concentration of salt increased the activity of antioxidant enzymes like superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX) and glutathione reductase in all chickpea genotypes but maximum activity was observed in salt-tolerant (SKUA-06 and SKUA-07) genotypes. Two genotypes of salt-tolerant and salt-sensitive varieties were analyzed further by real time PCR which revealed that the expression of SOD, APX and CAT genes were increased by NaCl in the salt-tolerant variety. The enhancement in tolerance against salt stress indicates that the genes involved in the antioxidative process are triggered by oxidative stress induced by environmental change. The results indicate that NaCl-induced oxidative stress hampers the normal functioning of the cell. The efficient antioxidants play a great role in mitigating the effect of NaCl stress in chickpea. This screening of NaCl-tolerant genotypes of chickpea can be performed on salt-affected land.


Cicer arietinum L. Growth Antioxidant enzymes Lipid peroxidation Oxidative stress Salinity 



The author gratefully acknowledges Hamdard National Foundation (HNF), New Delhi, India for providing financial assistance.


  1. Abu-Romman S, Shatnawi M (2011) Isolation and expression analysis of chloroplastic copper/zinc superoxide dismutase gene in barley. S Afr J Bot 77:328–334CrossRefGoogle Scholar
  2. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126CrossRefPubMedGoogle Scholar
  3. Agarwal S, Shaheen R (2007) Stimulation of antioxidant system and lipid peroxidation by abiotic stress in leaves of Momordica charantia. Braz J Plant Physiol 19:149–161CrossRefGoogle Scholar
  4. Ahmad P, Prasad MNV (2012a) Environmental adaptations and stress tolerance in plants in the era of climate change. Springer Science + Business Media, LLC, New YorkGoogle Scholar
  5. Ahmad P, Prasad MNV (2012b) Abiotic stress responses in Plants: metabolism, productivity and sustainability. Springer Science + Business Media, LLC, New YorkGoogle Scholar
  6. Ahmad P, Jhon R, Sarwat M, Umar S (2008a) Responses of proline, lipid peroxidation and antioxidative enzymes in two varieties of Pisum sativum L. under salt stress. Int J Plant Prod 2:353–366Google Scholar
  7. Ahmad P, Sarwat M, Sharma S (2008b) Reactive oxygen species, antioxidants and signaling in plants. J Plant Biol 51:167–173CrossRefGoogle Scholar
  8. Ahmad P, Jaleel CA, Salem MA, Nabi G, Sharma S (2010a) Roles of enzymatic and non-enzymatic antioxidants in plants during abiotic stress. Crit Rev Biotechnol 30:161–175CrossRefPubMedGoogle Scholar
  9. Ahmad P, Jaleel CA, Sharma S (2010b) Antioxidant defense system, lipid peroxidation, proline metabolizing enzymes, and biochemical activities in two Morus alba genotypes subjected to NaCl Stress. Russ J Plant Physiol 57:509–517CrossRefGoogle Scholar
  10. Ahmad P, Nabi G, Ashraf M (2011) Cadmium induced oxidative stress in mustard [Brassica juncea (L.) Czern. & Coss] plants can be alleviated by salicylic acid. S Afr J Bot 77:36–44CrossRefGoogle Scholar
  11. Ahmad P, Hakeem KR, Kumar A, Ashraf M, Akram NA (2012) Salt-induced changes in photosynthetic activity and oxidative defense system of three cultivars of mustard (Brassica juncea L.). Afr J Biotechnol 11(11):2694–2703Google Scholar
  12. Alscher RG, Erturk N, Heath LS (2002) Role of superoxide dismutases (SODs) in controlling oxidative stress in plants. J Exp Bot 53:1331–1341CrossRefPubMedGoogle Scholar
  13. Apel K, Hirt H (2004) Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Ann Rev Plant Biol 55:373–399CrossRefGoogle Scholar
  14. Arshi A, Ahmad A, Aref IM, Iqbal M (2012) Comparative studies on antioxidant enzyme action and ion accumulation in soybean cultivars under salinity stress. J Environ Biol 33:9–20PubMedGoogle Scholar
  15. Asada K (1992) Ascorbate peroxidase a hydrogen peroxide scavenging enzyme in plants. Plant Physiol 85:235–241CrossRefGoogle Scholar
  16. Ashraf M, Waheed A (1993) Responses of some genetically diverse lines of chickpea (Cicer arietinum L.) to salt. Plant Soil 154:257–266CrossRefGoogle Scholar
  17. Azooz MM, Shaddad MA, Abdel-Latef AA (2004) Leaf growth and K+/Na+ ratio as an indication of the salt tolerance of three sorghum cultivars grown under salinity stress and IAA treatment. Acta Agron Hung 52:287–296CrossRefGoogle Scholar
  18. Azooz MM, Youssef AM, Ahmad P (2011) Evaluation of salicylic acid (SA) application on growth, osmotic solutes and antioxidant enzyme activities on broad bean seedlings grown under diluted seawater. Int J Plant Physiol Biochem 3:253–264Google Scholar
  19. Bayer C, Fridovich I (1987) Superoxide dismutase: improved assays and applicable to acrylamide gels. Anal Biochem 44:276–287Google Scholar
  20. Bowler C, Van Montagu M, Inze D (1992) Superoxide dismutase and stress tolerance. Ann Rev Plant Physiol 43:83–116CrossRefGoogle Scholar
  21. Cushman JC, Michalowski CB, Bohnert HJ (1990) Developmental control of crassulacean acid metabolism inducibility by salt stress in the common ice plant. Plant Physiol 25:1137–1142CrossRefGoogle Scholar
  22. Dagar JC, Bhagwan H, Kumar Y (2004) Effect on growth performance and biochemical contents of Salvadora persica when irrigated with water of different salinity. Ind J Plant Physiol 9:234–238Google Scholar
  23. Dehghan G, Rezazadeh L, Habibi G (2011) Exogenous ascorbate improves antioxidant defense system and induces salinity tolerance in soybean seedlings. Acta Biol Szege 55:261–264Google Scholar
  24. Demiral T, Türkan I (2005) Comparative lipid peroxidation, antioxidant defense systems and proline content in roots of two rice cultivars differing in salt tolerance. Environ Exp Bot 53:247–257CrossRefGoogle Scholar
  25. Farhoudi R, Hussain M, Lee DJ (2012) Modulation of enzymatic antioxidants improves the salinity resistance in canola (Brassica napus). Int J Agric Biol 14:465–468Google Scholar
  26. Foyer CH, Halliwell B (1976) The presence of glutathione and glutathione reductase in Chloroplasts: a proposed role in ascorbic acid metabolism. Planta 133:21–25CrossRefGoogle Scholar
  27. Foyer CH, Descourvieres P, Kunert KJ (1994) Protection against oxygen radicals: an important defense mechanism studied in transgenic plants. Plant Cell Environ 17:507–523CrossRefGoogle Scholar
  28. Grattan SR, Grieve CM (1999) Salinity-mineral nutrient relations in horticulture crops. Sci Horticul 78:127–157CrossRefGoogle Scholar
  29. Gupta YP (1987) Studies on chemical and nutritional changes in Bengal gram (Cicer arietinum) during storage caused by the attack of pulse beetle (Callosobruchus maculatus Fab.). Plant Food Nutr 37:201–228CrossRefGoogle Scholar
  30. Heath RL, Packer L (1968) Photoperoxidation in isolated chloroplasts. I. Kinetics and stoichiometry of fatty acid peroxidation. Arch Biochem Biophys 125:189–198CrossRefPubMedGoogle Scholar
  31. Heidari M, Mesri F (2008) Salinity effects on compatible solutes, antioxidants enzymes and ion content in three wheat cultivars. Pak J Biol Sci 11:1385–1389CrossRefPubMedGoogle Scholar
  32. Hiscox JD, Israelstam GF (1979) A method for the extraction of chlorophyll from leaf tissue without maceration. Can J Bot 57:1332–1334CrossRefGoogle Scholar
  33. Hoagland DR, Arnon DI (1950) The water-culture method for growing plants without soil. California Agric Exp Stn Circ 347:1–32Google Scholar
  34. Hua XJ, Van de Cotte B, Van Montagu M, Verbruggen N (2001) The untranslated region the At- P5R gene is involved in both transcriptional and post transcriptional regulation. Plant J 26:157–169CrossRefPubMedGoogle Scholar
  35. Jaleel CA, Gopi R, Sankar B, Manivannan P, Kishore kumar A, Sridharan R, Panneerselvam R (2007) Studies on germination, seedling vigour, lipid peroxidation and proline metabolism in Catharanthus roseus seedlings under salt stress. S Afr J Bot 73:190–195CrossRefGoogle Scholar
  36. Katsuhara M, Otsuka T, Ezaki B (2005) Salt stress induced lipid peroxidation is reduced by glutathione S-transferase, but this reduction of lipid peroxides is not enough for a recovery of root growth in Arabidopsis. Plant Sci 169:369–373CrossRefGoogle Scholar
  37. Koca H, Ozdemir F, Turkan I (2006) Effect of salt stress on lipid peroxidation and superoxide dismutase and peroxidase activities of Lycopersicon esculentum and L. pennellii. Biol Plant 50:745–748CrossRefGoogle Scholar
  38. Koca M, Bor M, Ozdemir F, Turkan I (2007) The effect of salt stress on lipid peroxidation, antioxidative enzymes and proline content of sesame cultivars. Environ Exp Bot 60:344–351Google Scholar
  39. Koyro HW, Ahmad P, Geissler N (2012) Abiotic stress responses in plants: an overview. In: Ahmad P, Prasad MNV (eds) Environmental adaptations and stress tolerance of plants in the era of climate change. Springer Science + business media, New York, pp 1–28Google Scholar
  40. Levitt J (1980) Responses of plants to environmental stresses, vol 2. Academic Press, New YorkGoogle Scholar
  41. Macri F, Braidot E, Petrusa E, Vianello A (1994) Lipoxygenase activity associated to isolated soybean plasma membranes. Biochim Biophys Acta 1215:109–114CrossRefPubMedGoogle Scholar
  42. Masood A, Shah NA, Zeeshan M, Abraham G (2006) Differential response of antioxidant enzymes to salinity stress in two varieties of Azolla (Azolla pinnata and Azolla tilieuloides). Environ Exp Bot 58:216–222CrossRefGoogle Scholar
  43. Meloni DA, Oliva MA, Martinez CA, Cambraia J (2003) Photosynthesis and activity of superoxide dismutase, peroxidase and glutathione reductase in cotton under salt stress. Environ Exp Bot 49:69–76CrossRefGoogle Scholar
  44. Mengel K, Kirkby EA (2001) Principals of plant nutrition, 5th edn. Kluwer Academic Publishers, DordrechtCrossRefGoogle Scholar
  45. Metwally A, Safronova Belimov A, Dietz kj (2005) Genotypic variation of the response to cadmium toxicity in Pisum sativum L. J Exp Bot 56:167–178PubMedGoogle Scholar
  46. Mittal S, Kumari N, Sharma V (2012) Differential response of salt stress on Brassica juncea: photosynthetic performance, pigment, proline, D1 and antioxidant enzymes. Plant Physiol Biochem 54:17–26CrossRefPubMedGoogle Scholar
  47. Mittler R (2002) Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci 7:405–410CrossRefPubMedGoogle Scholar
  48. Nakano Y, Asada K (1981) Hydrogen peroxide is scavenged by ascorbate-specific peroxidases in spinach chloroplasts. Plant Cell Physiol 22:867–880Google Scholar
  49. Nazar R, Iqbal N, Syeed S, Khan NA (2011) Salicylic acid alleviates decreases in photosynthesis under saltstress by enhancing nitrogen and sulfur assimilation and antioxidant metabolism differentially in two mungbean cultivars. J Plant Physiol 168:807–815CrossRefPubMedGoogle Scholar
  50. Noctor G, Foyer CH (1998) Ascorbate and glutathione: keeping active oxygen under control. Annu Rev Plant Physiol Plant Mol Biol 49:249–279CrossRefPubMedGoogle Scholar
  51. Nounjan N, Nghia PT, Theerakulpisut P (2012) Exogenous proline and trehalose promote recovery of rice seedlings from salt-stress and differentially modulate antioxidant enzymes and expression of related genes. J Plant Physiol 169:596–604CrossRefPubMedGoogle Scholar
  52. Qilin D, Jin W, Bin F, Tingting L, Chen C, Honghui L, Shizhang D (2009) Molecular cloning and characterization of a new peroxidase gene (OvRCI) from Orychophragmus violaceus. Afr J Biotechnol 8:6511–6517Google Scholar
  53. Queiros F, Rodrigues JA, Almeida JM, Almeida DP, Fidalgo F (2011) Differential responses of the antioxidant defence system and ultrastructure in a salt-adapted potato cell line. Plant Physiol Biochem 49:1410–1419CrossRefPubMedGoogle Scholar
  54. Ramezani M, Seghatoleslami M, Mousavi G, Sayyari-Zahan MH (2012) Effect of salinity and foliar application of iron and zinc on yield and water use efficiency of Ajowan (Carum copticum). Intl J Agric Crop Sci 4:421–426Google Scholar
  55. Shi H, Lee BH, Wu SJ, Zhu JK (2003) over expression of a plasma membrane Na+/H+ antiporter gene improves salt tolerance in Arabidopsis thaliana. Nature Biotech 21:81–85CrossRefGoogle Scholar
  56. Siddiqi EH, Ashraf M, Al-Qurainy F, Akram NA (2011) Salt-induced modulation in inorganic nutrients, antioxidant enzymes, proline content and seed oil composition in safflower (Carthamus tinctorius L.). J Sci Food Agric 91:2785–2793CrossRefPubMedGoogle Scholar
  57. Smirnoff N (2000) Ascorbic acid metabolism and functions of a multifacet molecule. Curr Opin Plant Biol 3:229–235PubMedGoogle Scholar
  58. Tuteja N, Ahmad P, Panda BB, Tuteja R (2009) Genotoxic stress in plants: shedding light on DNA damage, repair and DNA repair helicases. Mutat Res 681:134–149CrossRefPubMedGoogle Scholar
  59. Van der Maesen LJG (1987) Origin, history and taxonomy of chickpea. In: Saxena MC, Singh KB (eds) The chickpea. CAB International, Oxon, pp 11–34Google Scholar
  60. Yamaguchi K, Mori H, Nishimura M (1995) A novel enzyme of ascorbate peroxidase localized on glyoxysomal and leaf peroxisomal membranes in pumpkin. Plant Cell Physiol 36:1157–1162PubMedGoogle Scholar
  61. Yang L, Bai X, Yang Y, Ahmad P, Yang Y, Hu X (2011) Deciphering the protective role of nitric oxide against salt stress at the physiological and proteomic levels in maize. J Proteom Res 10:4349–4364CrossRefGoogle Scholar
  62. Zahedi AM, Fazeli I, Zavareh M, Dorry H, Gerayeli N (2012) Evaluation of the sensitive components in seedling growth of common bean (Phaseolus vulgaris L.) affected by salinity. Asian J Crop Sci 4:159–164CrossRefGoogle Scholar
  63. Zheng YH, Xu XB, Wang MY, Zheng XH, Li ZJ, Jiang GM (2009) Responses of salt-tolerant and intolerant wheat genotypes to sodium chloride: photosynthesis, antioxidants activities, and yield. Photosynthetica 47:87–94CrossRefGoogle Scholar

Copyright information

© Franciszek Górski Institute of Plant Physiology, Polish Academy of Sciences, Kraków 2012

Authors and Affiliations

  • Saiema Rasool
    • 1
  • Altaf Ahmad
    • 1
  • T. O. Siddiqi
    • 1
  • Parvaiz Ahmad
    • 2
  1. 1.Department of BotanyJamia HamdardNew DelhiIndia
  2. 2.Department of Botany, A.S. CollegeUniversity of KashmirSrinagarIndia

Personalised recommendations