Journal of Crop Science and Biotechnology

, Volume 17, Issue 1, pp 11–20 | Cite as

Growth, physiological, and biochemical responses in relation to salinity tolerance for In Vitro selection in oil seed crop Guizotia abyssinica Cass.

  • Savaliram Goga Ghane
  • Vinayak Haribhau Lokhande
  • Tukaram Dayaram NikamEmail author
Research Article


The calli cultures of Guizotia abyssinica (niger) cultivars IGP 76 and GA 10 were exposed to different levels of salt treatments (0, 30, 60, and 90 mM NaCl), in order to evaluate growth, physiological, and biochemical responses. A significant decrease in relative growth rate and tissue water content of GA 10 calli than IGP 76 under salt-stress conditions was associated with higher sodium ion accumulation. Osmotic adjustment revealed by the osmolytes (proline, glycine betaine, and total soluble sugars) accumulation was significantly higher in IGP 76 salt-stressed calli as compared to GA 10. The sustained growth and better survival of IGP 76 calli was correlated with lower malondialdehyde content and increased superoxide dismutase, ascorbate peroxidase, and catalase activities and higher α-tocopherol content in comparison to GA 10. The higher osmolytes accumulation and presence of better antioxidant system suggested superior adaptation of IGP 76 calli on salt-containing medium for prolonged periods in comparison to GA 10. The regeneration frequency, organogenesis, and acclimatization response of the plants derived from salt-adapted calli was comparatively lower than the plants derived from control calli of IGP 76. The growth, physiological, and biochemical characterization of the salt-tolerant regenerated plants exposed to stepwise long-term 90 mM NaCl treatment revealed no significant changes in comparison to the control. Thus, our results suggests the development of an efficient protocol for in vitro selection and production of salt-tolerant plants in self-incompatible crop, niger, and an alternative to traditional breeding programs to increase the abiotic stress tolerance.

Key words

antioxidant enzymes callus culture Guizotia abyssinica in vitro selection osmolytes salt tolerant plant 


Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.


  1. Alvarez I, Tomaro ML, Benavides MP. 2003. Changes in polyamines, proline and ethylene in sunflower calluses treated with NaCl. Plant Cell Tiss. Org. Cult. 74: 51–59CrossRefGoogle Scholar
  2. Ashraf M. 2009. Biotechnological approach of improving plant salt tolerance using antioxidants as markers. Biotech. Adv. 27: 84–93CrossRefGoogle Scholar
  3. Ashraf M, Foolad MR. 2007. Roles of glycine betaine and proline in improving plant abiotic stress resistance. Environ. Expt. Bot. 59: 206–216CrossRefGoogle Scholar
  4. Attipali RR, Kolluru VC, Munusamy V. 2004. Drought induced responses of photosynthesis and antioxidant metabolism in higher plants. J. Plant Physiol. 161: 1189–1202CrossRefGoogle Scholar
  5. Basu S, Gangopadhyay G, Mukherjee BB. 2002. Salt tolerance in rice in vitro: Implication of accumulation of Na+, K+ and proline. Plant Cell Tiss. Org. Cult. 69: 55–64CrossRefGoogle Scholar
  6. Bates LS, Waldren RP, Teare ID. 1973. Rapid determination of free proline for water stress studies. Plant Soil 39:205–208CrossRefGoogle Scholar
  7. Becana M, Moran JF, Iturbe-Ormaetxe I. 1998. Iron-dependent oxygen free radical generation in plants subjected to environmental stress: toxicity and antioxidant protection. Plant Soil 201: 137–147CrossRefGoogle Scholar
  8. Cakmak I, Marschner H. 1992. Magnesium deficiency and high light intensity enhance activities of superoxide dismutase, ascorbate peroxidase, and glutathione reductase in bean leaves. Plant Physiol. 98: 1222–1227PubMedCentralPubMedCrossRefGoogle Scholar
  9. Cushman JC, De Rocher EJ, Bohnert HJ. 1990. Gene expression during adaptation to salt stress. In F Katterman, Eds, Environmental Injury to Plants. Academic Press, Inc., USA, pp 173–203Google Scholar
  10. Davenport SB, Gallego SM, Benavides MP, Tomaro ML. 2003. Behaviour of antioxidant defense system in the adaptive response to salt stress in Helianthus annuus L. cells. Plant Grow. Regul. 40: 81–88CrossRefGoogle Scholar
  11. Demiral T, Turkan I. 2005. Comparative lipid peroxidation, antioxidant defense systems and proline content in roots of two rice cultivars differing in salt tolerance. Environ. Expt. Bot. 53: 247–257CrossRefGoogle Scholar
  12. Elkahoui S, Hernandez JA, Abdelly C, Ghrir R, Limam F. 2005. Effects of salt on lipid peroxidation and antioxidant enzyme activities of Catharanthus roseus suspension cells. Plant Sci. 168: 607–613CrossRefGoogle Scholar
  13. Getinet A, Sharma SM. 1996. Niger [Guizotia abyssinica (L. f.) Cass.] IBPGRI, Rome.Google Scholar
  14. Grieve CM, Grattan SR. 1983. Rapid assay for determination of water soluble quaternary ammonium compounds. Plant Soil 70: 303–307CrossRefGoogle Scholar
  15. Heath RL, Packer L. 1968. Photoperoxidation in isolated chloroplasts. Kinetics and stoichiometry of fatty acid peroxidation. Arch. Biochem. Biophy. 125: 189–198CrossRefGoogle Scholar
  16. Hossain Z, Mandal AKA, Datta SK, Biswas AK. 2007. Development of NaCl-tolerant line in Chrysanthemum morifolium Ramat., through shoot organogenesis of selected callus line. J. Biotechnol. 129: 658–667PubMedCrossRefGoogle Scholar
  17. Jain M. 2001. Tissue culture-derived variation in crop improvement. Euphytica 118: 153–166CrossRefGoogle Scholar
  18. Jain M, Mathur G, Koul S, Sarin NB. 2001. Ameliorative effects of proline on salt stress-induced lipid peroxidation in cell lines of groundnut (Arachis hypogaea L.) Plant Cell Rep. 20: 463–468CrossRefGoogle Scholar
  19. Joshi, PK, Saxena SC, Arora S. 2011. Characterization of Brassica juncea antioxidant potential under salinity stress. Acta Physiol. Plant. 33: 811–822CrossRefGoogle Scholar
  20. Leone A, Costa A, Tucci M, Grillo S. 1994. Adaptation versus shock response to polyethylene glycol-induced low water potential in cultured potato cells. Physiol. Plant. 92: 21–30CrossRefGoogle Scholar
  21. Lichtenthaler HK, Buschmann C. 2001. Chlorophylls and carotenoids: measurement and characterization by UVVIS spectroscopy. In Current Protocols in Food Analytical Chemistry, John Wiley and Sons, New York, pp F4.3.1–F4.3.8Google Scholar
  22. Liu T, Staden JV. 2000. Selection and characterization of sodiumchloride-tolerantcallus of Glycine max (L.) Merr cv. Acme. Plant Grow. Regul. 31: 195–207CrossRefGoogle Scholar
  23. Lokhande VH, Nikam TD, Suprasanna P. 2010. Biochemical, physiological and growth changes in response to salinity in callus cultures of Sesuvium portulacastrum L. Plant Cell Tiss. Org. Cult. 102: 17–25CrossRefGoogle Scholar
  24. Lowry OH, Rosebrough HJ, Farr AL, Randall RJ. 1951. Protein measurement with folin phenol reagent. J. Biol. Chem. 193: 265–275PubMedGoogle Scholar
  25. Miller G, Honig A, Stein H, Suzuki N, Mittler R, Zilberstein A. 2009. Unraveling Δ1-pyrroline-5-carboxylate-proline cycle in plants by uncoupled expression of proline oxidation enzymes. J. Biol. Chem. 284(39): 26482–26492PubMedCentralPubMedCrossRefGoogle Scholar
  26. Mittler R. 2002. Oxidative stress, antioxidants and stress tolerance. Trends Plant Sci. 7: 405–410PubMedCrossRefGoogle Scholar
  27. Munns R, Tester M. 2008. Mechanisms of salinity tolerance. Ann. Rev. Plant Biol. 59: 651–681CrossRefGoogle Scholar
  28. Murashige T, Skoog F. 1962. A revised medium for rapid growth and bioassays with tobacco tissue culture. Physiol. Plant. 15: 473–497CrossRefGoogle Scholar
  29. Nakano Y, Asada K. 1981. Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts. Plant Cell Physiol. 22: 867–880Google Scholar
  30. Nikam TD, Shitole MG. 1997. In vitro plant regeneration from callus of niger (Guizotia abyssinica Cass.) cv. Sahyadri. Plant Cell Rep. 17: 155–158CrossRefGoogle Scholar
  31. Panfili G, Manzi P, Pizzoferrato L. 1994. High-performance liquid chromatographic method for the simultaneous determination of tocopherols, carotenes, and retinol and its geometric isomers in Italian cheeses. Analyst 119: 1161–1165PubMedCrossRefGoogle Scholar
  32. Parida AK, Das AB. 2005. Salt tolerance and salinity effects on plants: a review. Ecotoxicol. Environ. Safety 60: 324–349PubMedCrossRefGoogle Scholar
  33. Patade VY, Suprasanna P, Bapat VA. 2008. Effects of salt stress in relation to osmotic adjustment on sugarcane (Saccharum officinarum L.) callus cultures. Plant Grow. Regul. 55(3): 169–173CrossRefGoogle Scholar
  34. Purohit M, Srivastava S, Srivastava PS. 1998. Stress tolerant plants through tissue culture. In PS Srivastava, ed, Plant Tissue Culture and Molecular Biology: Application and Prospects, Narosa Publishing House, New Delhi, India, pp 554–578Google Scholar
  35. Queiros F, Fidalgo F, Santos I, Salema R. 2007. In vitro selection of salt tolerant cell lines in Solanum tuberosum L. Biol. Plant. 51: 728–734CrossRefGoogle Scholar
  36. Rai MK, Kalia RK, Singh R, Gangola MP, Dhawan AK. 2011. Developing stress tolerant plants through in vitro selection—An overview of the recent progress. Environ. Expt. Bot. 71: 89–98CrossRefGoogle Scholar
  37. Ramadan MF, Morsel JT. 2002. Proximate neutral lipid composition of niger. Czech J. Food Sci. 20: 98–104Google Scholar
  38. Sheekh-El MM, Omar HH. 2002. Effect of high salt stress on growth and fatty acids content of the unicellular green algae Chlorella vulgaris. Amer. J. Microbiol. 55: 181–191Google Scholar
  39. Watanabe S, Kojima K, Ide Y, Sasaki S. 2000. Effects of saline and osmotic stress on proline and sugar accumulation in Populus euphraticain vitro. Plant Cell Tiss. Org. Cult. 63: 199–206CrossRefGoogle Scholar
  40. Yasar F, Ellialtioglu S, Kusvuran S. 2006. Ion and lipid peroxide content in sensitive and tolerant eggplant callus cultured under salt stress. Europ. J. Hortic. Sci. 71(4): 169–172Google Scholar
  41. Yokoi S, Bressan RA, Hasegawa PM. 2002. The Japan International Centre for Agricultural Sciences (JIRCAS) Working Report No. 23. In M Iwanaga, Eds., Genetic engineering of crop plants for abiotic stress. Salt stress tolerance of plants. Japan International Centre for Agricultural Sciences, Tsukuba, pp 25–33Google Scholar
  42. Zhang F, Yang YL, He WL, Zhao X, Zhang LX. 2004. Effects of salinity on growth and compatible solutes of callus induced from Populus euphratica. In vitro Cell. Dev. Biol. — Plant 40: 491–494CrossRefGoogle Scholar
  43. Zhao X, Tan HJ, Liu YB, Li XR, Chen GX. 2009. Effect of salt stress on growth and osmotic regulation in Thellungiella and Arabidopsis callus. Plant Cell Tiss. Org. Cult. 98(1): 97–103CrossRefGoogle Scholar

Copyright information

© Korean Society of Crop Science and Springer Science+Business Media Dordrecht 2013

Authors and Affiliations

  • Savaliram Goga Ghane
    • 1
  • Vinayak Haribhau Lokhande
    • 2
  • Tukaram Dayaram Nikam
    • 3
    Email author
  1. 1.Department of BotanyShivaji UniversityKolhapurIndia
  2. 2.Department of BotanyShri Shiv Chhatrapati CollegeBodkenagarMaharashtra, India
  3. 3.Department of BotanyUniversity of PunePuneIndia

Personalised recommendations