Effect of exogenous application of IAA and GA3 on growth, protein content, and antioxidant enzymes of Solanum tuberosum L. grown in vitro under salt stress

  • Arifa Khalid
  • Faheem AftabEmail author
Plant Tissue Culture


Indole-3-acetic acid (IAA) and gibberellic acid (GA3) are essential for the growth and development of plants. In the present study, the ameliorative potential of these phytohormones on growth, protein content, and antioxidant enzymes was investigated in in vitro-grown Solanum tuberosum L. cultivars ‘Cardinal’ and ‘Desiree’ under salt stress. A 4 × 3 factorial combination of 0, 40, 60, or 80 mM NaCl with 0, 7, or 14 μM IAA, or 0, 14, or 21 μM GA3, were added to Murashige and Skoog (MS) basal medium, followed by inoculation of nodal explants or callus cultures. The data for root and shoot number and length, number of nodes and leaves, fresh weight of plants, increase or decrease in fresh weight of callus cultures, total soluble protein, and superoxide dismutase (SOD) and peroxidase (POD) activities were recorded after 30 d. The growth of both callus cultures and nodal explants subjected to NaCl stress was substantially reduced compared with the control. Both IAA and GA3 successfully alleviated the harmful effects of salt stress on all of the growth parameters studied. Salt stress resulted in decreased protein content, which increased when the media also contained phytohormones. The activities of SOD and POD were increased with either IAA or GA3 under NaCl stress. Therefore, the exogenous application of both IAA and GA3 not only played a positive role in terms of in vitro potato growth but also significantly affected the biochemical parameters tested.


Antioxidants GA3 IAA Protein Salt stress 



We are grateful to the anonymous reviewers for their excellent reviews and feedback. We also thank the Copy Editor and the Editor in Chief for their contribution to enhance the outlook of this manuscript a great deal.

Funding information

We thank Higher Education Commission Pakistan for providing research funds in the form of the Indigenous 5000 PhD Fellowship (106-1137-BM6-088) to Arifa Khalid.


  1. Agastian P, Kingsley SJ, Vivekanandan M (2000) Effect of salinity on photosynthesis and biochemical characteristics in mulberry genotypes. Photosynthetica 38:287–290CrossRefGoogle Scholar
  2. Aghaei K, Ehsanpour AA, Komatsu S (2009) Potato responds to salt stress by increased activity of antioxidant enzymes. J Integr Plant Biol 51:1095–1103PubMedCrossRefPubMedCentralGoogle Scholar
  3. Ahmad P, Azooz, MM, Prasad MNV (Eds.). (2012) Ecophysiology and responses of plants under salt stress. Springer Science & Business MediaGoogle Scholar
  4. Akbari GA, Arab SM, Alikhani HA, Allakdadi I, Arzanesh MH (2007) Isolation and selection of indigenous Azospirillum spp. and the IAA of superior strains effects on wheat roots. W J Agri Sci 3:523–529Google Scholar
  5. Al-Hakimi AMA, Hamada AM (2001) Counteraction of salinity stress on wheat plants by grain soaking in ascorbic acid, thiamin or sodium salicylate. Biol Plant 44:253–261CrossRefGoogle Scholar
  6. Amzallag GN, Lener HR, Poljakoff-Mayber A (1990) Exogenous ABA as a modulator of the response of Sorghum to high salinity. J Exp Bot 541:1529–1534CrossRefGoogle Scholar
  7. Anuradha S, Rao SSR (2007) The effect of brassinosteroids on radish (Raphanus sativus L.) seedlings growing under cadmium stress. Plant Soil Environ 53:465–472CrossRefGoogle Scholar
  8. Arteca RN (1996) Plant growth substances. Chapman and Hall, New York, pp 286–287Google Scholar
  9. Ashraf M, Fooland MR (2005) Pre sowing seed treatment—a shotgun approach to improve germination, plant growth and crop yield under saline and non-saline conditions. Adv Agron 88:223–271CrossRefGoogle Scholar
  10. Ashraf M, Harris PJC (2004) Potential biochemical indicators of salinity tolerance in plants. Plant Sci 166:3–16CrossRefGoogle Scholar
  11. Ayala-Astorga GI, Alcaraz-Meléndez L (2010) Salinity effects on protein content, lipid peroxidation, pigments, and proline in Paulownia imperialis (Siebold & Zuccarini) and Paulownia fortunei (Seemann & Hemsley) grown in vitro. Electron J Biotechnol 13:13–14CrossRefGoogle Scholar
  12. Borsani O, Valpuesta V, Botella MA (2003) Developing salt tolerant plants in a new century: a molecular biology approach. Plant Cell Tissue Organ Cult 73:101–115CrossRefGoogle Scholar
  13. Chakrabarti N, Mukherji S (2003) Effect of phytohormone pretreatment on nitrogen metabolism in Vigna radiata under salt stress. Biol Plant 46:63–66CrossRefGoogle Scholar
  14. Chauhan JS, Tomar YK, Singh NI, Ali S, Debrati (2009) Effect of growth hormones on seed germination and seedling growth of black gram and horse gram. J Am Sci 5:79–78Google Scholar
  15. Claussen M, Lüthen H, Blatt MR, Böttger M (1997) Auxin induced growth and its linkage to potassium channels. Planta 201:227–234CrossRefGoogle Scholar
  16. Dalton FN, Maggio A, Piccinni G (2000) Simulation of shoot chloride accumulation: separation of physical and biochemical processes governing plant salt tolerance. Plant Soil 219:1–11CrossRefGoogle Scholar
  17. Datta KS, Varma SK, Angrish R, Kumar B, Kumari P (1998) Alleviation of salt stress by plant growth regulators in Triticum aestivum L. Biol Plant 42:269–275Google Scholar
  18. Egamberdieva D (2009) Alleviation of salt stress by plant growth regulators and IAA producing bacteria in wheat. Acta Physiol Plant 31:861–864CrossRefGoogle Scholar
  19. Esfandiari E, Shekari F, Shekari F, Esfandiari M (2007) The effects of salt stress on antioxidant enzymes’ activity and lipid peroxidation on the wheat seedlings. Notulae Botanicae Horti Agrobotani Cluj-Napoca 35:58–56Google Scholar
  20. FAO (2008) Hidden treasure. International year of potato, 2008. Food and agriculture organization of the United Nations.
  21. Fidalgo F, Santos A, Santos I, Salema R (2004) Effect of long-term salt stress on antioxidant defense system, leaf water relations and chloroplast ultra-structure of potato plant. Ann Appl Biol 145:185–192CrossRefGoogle Scholar
  22. Foyer CH, Shigeoka S (2011) Understanding oxidative stress and antioxidant functions to enhance photosynthesis. Plant Physiol 155:93–100PubMedCrossRefPubMedCentralGoogle Scholar
  23. Gill SS, Tuteja N (2010) Reactive oxygen species and antioxidant machinery in abiotic stress tolerance in crop plants. Plant Physiol Biochem 48:909–930PubMedPubMedCentralCrossRefGoogle Scholar
  24. Hamayun M, Khan SA, Khan AL, Shin JH, Ahmad B, Shin DH, Lee IJ (2010) Exogenous gibberellic acid reprograms soybean to higher growth and salt stress tolerance. J Agr Food Chem 58:7226–7232CrossRefGoogle Scholar
  25. HanumanthaRao B, Nair RM, Nayyar H (2016) Salinity and high temperature tolerance in mungbean [Vigna radiata (L.) Wilczek] from a physiological perspective. Frontiers. Plant Sci 7:957Google Scholar
  26. Ilias I, Ouzounidou G, Giannakoula A, Papdopoulou P (2007) Effect of gibberellic acid and prohexadione-calcium on growth, chlorophyll fluorescence and quality of okra plant. Biol Plant 51:575–578CrossRefGoogle Scholar
  27. Karimi G, Ghorbanli M, Heidari RA, Assareh MH (2005) The effects of NaCl on growth, water relations, osmolytes and ion content in Kochia prostrate. Biol Plant 49:301–334CrossRefGoogle Scholar
  28. Kaur H, Gupta N (2018) Ameliorative effect of proline and ascorbic acid on seed germination and vigour parameters of tomato (Solanum tuberosum L.) under salt stress. Int J Curr Microbiol App Sci 7:3523–3532CrossRefGoogle Scholar
  29. Kaur S, Gupta AK, Kaur N (2000) Effects of GA3, kinetin and indole acetic acid on carbohydrate metabolim in chickpea seedlings germinating under water stress. Plant Growth Regul 30:61–70CrossRefGoogle Scholar
  30. Khalid A (2017) Effect of exogenous application of IAA, BRs and GA3 on growth, protein contents and antioxidative enzyme activities in Solanum tuberosum L. under salt stress. Doctoral dissertation, University of the Punjab, LahoreGoogle Scholar
  31. Khalid A, Aftab F (2016) Effect of exogenous application of 24-epibrassinolide on growth, protein contents and antioxidant enzyme activities of Solanum tuberosum L. under salt stress. In Vitro Cell Dev Biol-Plant 52:81–91CrossRefGoogle Scholar
  32. Kim S, Kang JY, Cho DI, Park JH, Kim SY (2004) ABF2, an ABRE-binding bZIP factor, is an essential component of glucose signaling and its overexpression affects multiple stress tolerance. Plant Physiol 40:75–87Google Scholar
  33. Li Y (2009) Effects of NaCl stress on antioxidant enzymes of Glycine Soja sieb. Pak J Biol Sci 12:510–513PubMedCrossRefPubMedCentralGoogle Scholar
  34. Liu T, Van-Staden J (1999) Selection and characterization of sodium chloridetolerant callus of Glycine max (L). Merrcb Acme 31:195–207Google Scholar
  35. Manchandia AM, Banks SW, Gossett DR, Bellaire BA, Lucas MC, Millhollon EP (1999) The influence of α-amanitin on the NaCl-induced up-regulation of antioxidant enzyme activity in cotton callus tissue. Free Radic Res 30:429–438PubMedCrossRefPubMedCentralGoogle Scholar
  36. Maral J, Puget K, Micheson AM (1977) Comparative study of superoxide dismutase, catalase, and glutathione peroxidase levels in erythrocytes of different animals. Biochem Biophys Res Commun 77:1525–1535PubMedCrossRefPubMedCentralGoogle Scholar
  37. Meloni DA, Oliva MA, Ruiz HA, Martinez CA (2001) Contribution of proline and inorganic solutes to osmotic adjustment in cotton under salt stress. J Plant Nutr 24:599–612CrossRefGoogle Scholar
  38. Molassiotis AN, Sotiropoulos T, Tanou G, Kofidis G, Diamantidis G, Therios I (2006) Antioxidant and anatomical responses in shoot culture of the apple rootstock MM 106 treated with NaCl, KCl, mannitiol or sorbitol. Biol Plant 50:61–68CrossRefGoogle Scholar
  39. Munns R (2002) Comparative physiology of salt and water stress. Plant Cell Environ 25:239–250PubMedPubMedCentralCrossRefGoogle Scholar
  40. Murashige T, Skoog F (1962) A revised medium for a rapid growth and bio assays with tobacco tissue cultures. Physiol Plant 15:473–497CrossRefGoogle Scholar
  41. Ochatt SJ, Marconi PL, Radice S, Arnozis PA, Caso OH (1999) In vitro recurrent selection of potato: production and characterization of salt-tolerant cell lines and plants. Plant Cell Tissue Organ Cult 55:1–8CrossRefGoogle Scholar
  42. Parida AK, Das AB (2005) Salt tolerance and salinity effects on plants: a review. Ecotox Environ Safe 60:324–349CrossRefGoogle Scholar
  43. Parida AK, Das AB, Mittra B, Mohanty P (2004) Salt-stress induced alterations in proteins profile and protease activity in the mangrove. Bruguiera parviflora Zeitschrift für Naturforschung, Z Naturforsch 59:408–414Google Scholar
  44. Qasim A, Athar HR, Ahraf M (2006) Influence of exogenously applied brassinosteroids on the mineral nutrient status of two cultivars grown under saline conditions. Pak J Bot 38:1621–1632Google Scholar
  45. Racusen D, Foote M (1965) Protein synthesis in dark grown bean leaves. Can J Bot 43:817–824CrossRefGoogle Scholar
  46. Racusen D, Johnstone DB (1961) Estimation of protein in cellular material. Nature 191:292–493CrossRefGoogle Scholar
  47. Rahman A, Hosokawa S, Oono Y, Amakawa T, Goto N, Tsurumi S (2002) Auxin and ethylene response interactions during Arabidopsis root hair development dissected by auxin influx modulators. Plant Physiol 130:1908–1917PubMedPubMedCentralCrossRefGoogle Scholar
  48. Rahnama H, Ebrahimzadeh H (2005) The effect of NaCl on antioxidant enzyme activities in potato seedlings. Biol Plant 49:93–97CrossRefGoogle Scholar
  49. Robinson T (1979) The determination of proteins in plant extracts that contain polyphenols. Plant Sci Lett 15:211–216CrossRefGoogle Scholar
  50. Senthila A, Djanaguiraman M, Vijayalkashmi C (2005) Influence of seed treatment of growth regulators on some enzyme activities in groundnut under salinity. Agric Trop Subtrop 38:88–90Google Scholar
  51. Shah SH, Ahmad I, Samiullah (2007) Responses of Nigella sativa to foliar application of gibberellic acid and kinetin. Biol Plant 51:563–566CrossRefGoogle Scholar
  52. Shakirova FM, Sakhabutdinova AR, Bezukova MV, Fatkhutdinova RA, Fatkhutdinova DR (2003) Changes in the hormonal status of wheat seedlings induced by salicylic acid and salinity. Plant Sci 164:317–322CrossRefGoogle Scholar
  53. Singh NK, Bracken CA, Hasegawa PM, Handa AK, Buckel S, Hermodson MA, Pfankoch F, Regnier FE, Bressan RA (1987) Characterization of osmotin. A thaumatin-like protein associated with osmotic adjustment in plant cells. Plant Physiol 85:529–536PubMedPubMedCentralCrossRefGoogle Scholar
  54. Tal M (1983) Selection for stress tolerance. In: Evans DA, Sharp WR, Ammirato PV, Ymada Y (eds) Hand book of plant cell culture, vol 1. McMillan, London, pp 461–488Google Scholar
  55. Tanveer M (2019) Role of 24-Epibrassinolide in inducing thermo-tolerance in plants. J Plant Growth Regul 38:945–955CrossRefGoogle Scholar
  56. Teixeira J, Pereira S (2007) High salinity and drought act on an organ-dependent manner on potato glutamine synthetase expression and accumulation. Environ Exp Bot 60:121–126CrossRefGoogle Scholar
  57. Tuna AL, Kaya C, Dikilitas M, Higgs D (2008) The combined effects of gibberellic acid and salinity on some antioxidant enzyme activities, plant growth parameters and nutritional status in maize plants. Environ Exp Bot 62:1–9CrossRefGoogle Scholar
  58. Veselov DS, Sabirzhanova IB, Sabirzhanov BE, Chemeris AV (2008) Changes in expansin gene expression, IAA content, and extension growth of leaf cells in maize plants subjected to salinity. Russ. J. Plant Physiol 55:101–106Google Scholar
  59. Wang W, Vinocur B, Altman A (2003) Plant responses to drought, salinity and extreme temperatures: toward genetic engineering for stress tolerance. Planta 218:1–14PubMedCrossRefPubMedCentralGoogle Scholar
  60. Wang Y, Nil N (2000) Changes in chlorophyll, ribolose bisphasphate arboxylaseoxygenase, glycine betaine contents, photosynthesis and transpiration in Amaranthus tricolor leaves during salt stress. J Hortic Sci Biotechnol 75:623–627CrossRefGoogle Scholar
  61. Wen FP, Zhang ZH, Bai T, Xu Q, Pan YH (2010) Proteomics reveals the effects of gibberellic acid (GA3) on salt-stressed rice (Oryza sativa L.) shoots. Plant Sci 178:170–175CrossRefGoogle Scholar
  62. Yurekli F, Porgali ZB, Turkan I (2004) Variation in abscisic acid, indole-3-acetic acid, gibberellic acid and zeatin concentrations in two bean species subjected to salt stress. Acta Biol Cracov 46:201–212Google Scholar
  63. Zhu JK (2001) Overexpression of delta-pyrroline-5-carboxylate synthetase gene and analysis of tolerance to water and salt stress in transgenic rice. Trends Plant Sci 6:66–72PubMedPubMedCentralCrossRefGoogle Scholar

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© The Society for In Vitro Biology 2020

Authors and Affiliations

  1. 1.Department of BotanyUniversity of the PunjabLahorePakistan

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