Adaptive Responses of Carum copticum to in vitro Salt Stress

Authors: R. Razavizadeh, F. Adabavazeh, M. Rezaee Chermahini

Abstract:

Salinity is one of the most widespread agricultural problems in arid and semi-arid areas that limits the plant growth and crop productivity. In this study, the salt stress effects on protein, reducing sugar, proline contents and antioxidant enzymes activities of Carum copticum L. under in vitro conditions were studied. Seeds of C. copticum were cultured in Murashige and Skoog (MS) medium containing 0, 25, 50, 100 and 150 mM NaCl and calli were cultured in MS medium containing 1 μM 2, 4-dichlorophenoxyacetic acid, 4 μM benzyl amino purine and different levels of NaCl (0, 25, 50, 100 and 150 mM). After NaCl treatment for 28 days, the proline and reducing sugar contents of shoots, roots and calli increased significantly in relation to the severity of the salt stress. The highest amount of proline and carbohydrate were observed at 150 and 100 mM NaCl, respectively. The reducing sugar accumulation in shoots was the highest as compared to roots, whereas, proline contents did not show any significant difference in roots and shoots under salt stress. The results showed significant reduction of protein contents in seedlings and calli. Based on these results, proteins extracted from the shoots, roots and calli of C. copticum treated with 150 mM NaCl showed the lowest contents. The positive relationships were observed between activity of antioxidant enzymes and the increase in stress levels. Catalase, ascorbate peroxidase and superoxide dismutase activity increased significantly under salt concentrations in comparison to the control. These results suggest that the accumulation of proline and sugars, and activation of antioxidant enzymes play adaptive roles in the adaptation of seedlings and callus of C. copticum to saline conditions.

Keywords: Antioxidant enzymes, Carum copticum, organic solutes, salt stress.

Digital Object Identifier (DOI): doi.org/10.5281/zenodo.1128905

References:

[1] R. Munns, R. A. James, A. Lauchli, “Approaches to increasing the salt tolerance of wheat and other cereals,” J. Exp. Bot., vol. 57, pp. 1-19, 2006.
[2] A. Zabalza, S. Gaston, L. M. Sandalio, L. A. Del Rio and M. Royuela, “Oxidative stress is not related to the mode of action of herbicides that inhibit acetolactate synthase,” Environ. Exper. Bot., vol. 59, pp. 150-159, 2007.
[3] I. M. Moller, P. E. Jensen and A. Hansson, “Oxidative modifications to cellular components in plants,” Annu. Rev. Plant Biol., vol. 58, pp. 459-481, 2007.
[4] N. Smirnoff, “The role of active oxygen in the response of plants to water deficit and desiccation,” New Phytologist., vol. 125, pp. 27–58, 1993.
[5] R. R. Attipali, V. C. Kolluru, V. Munusamy, “Drought induced responses of photosynthesis and antioxidant metabolism in higher plants,” J. Plant Physiol., vol. 161, pp. 1189-1202, 2004.
[6] A. N. Misra, A. K. Biswal and M. Misra, “Physiological, biochemical and molecular aspects of water stress in plants, and their biotechnological applications,” Proc. Nat Acad. Sci., vol. 72, pp. 115-134, 2002.
[7] M. M. Zarshenas, M. Moein, M. Mohammadi Samani and P. Petramfar, “An overview on Ajwain (Trachyspermum ammi) pharmacological effects; modern and traditional,” J. Nat Remedies, vol. 14, , pp. 98-1052013.
[8] K. Cimanga, K. Kambu, L. Tona, S. Apers, T. De Bruyne and N. Hermans, “Correlation between chemical composition and antibacterial activity of essential oils of some aromatic medicinal plants growing in the Democratic Republic of Congo,” J. Ethnopharmacol., vol. 79, pp. 213-220, 2002.
[9] M. Shojaaddini, S. Moharramipour and B. Z. Sahaf, “Fumigant toxicity of essential oil from Carum copticum against Indian meal moth, Plodia interpunctella,” J. Plant Prot. Res., vol. 48, pp. 411-9, 2008.
[10] T. Murashige and F. Skoog, “A revised medium for rapid growth and bioassays with tobacco tissue cultures,” Plant Physiol., vol. 15, pp. 473-497, 1962.
[11] M. Somogyi-Nelson, “Notes on sugar determination,” J. Biol. Chem., vol. 195, pp. 19-29, 1952.
[12] L. S. Bates, R. P. Waldern and I.D. Teare, “Rapid determination of free proline for water stress studies,” J. Plant Soil., vol. 39, no. 1, pp. 205-207, 1973.
[13] M. M. Bradford, “A rapid and sensitive method for the quantification of microgram quantities of protein utilizing the principle of protein-dye binding,” Annu. Rev. Biochem., vol. 72, pp. 248-254, 1976.
[14] C. N. Giannotolitis and S.K. Ries, “Superoxide dismutase: II. Purification and quantitative relationship with water-soluble protein in seedling,” Plant Physiol, vol. 59, pp. 315-318, 1997.
[15] H. Aebi, “Catalases. In: Bergmeyer HU (ed.), Methods of enzymatic analysis,” Academic Press, New York, pp. 673-684, 1974.
[16] Y. Nakano and K. “Asada Hydrogen peroxide is scavenged by ascorbate-specific peroxidase in spinach chloroplasts,” Plant Cell Physiol., vol. 22, pp. 867-880, 1981.
[17] A. Maggio, S. Miyazakil, P. Veronese, T. Tomomichi Fujita, J. Ibeas, B. Damszl, M. L. Narasimhan, P. M. Hasegawal, R. J. Joly and R. A. Bressan, “Does proline accumulation play an active role in stress-induced growth reduction?” Plant J., vol. 31, pp. 699-712, 2002.
[18] N. Verbrugg and C. Hermans, “Proline accumulation in plants: a review,” Amino Acids., vol. 35, pp. 753-759, 2008.
[19] R. C. Ackerson, “Osmoregulation of cotton in response to water stress III. Effect of pjosphorus fertility,” J. Plant Physiol., vol. 77, pp. 309-312, 1985.
[20] D. K. Srivastava, V. K. Gupta and D.R. Sharma, “In vitro selection and characterization of water stress tolerance callus cultures of tomato (Lycopersicum esculentum L.),” Indian J. Plant Physiol., vol. 2, pp. 99-104, 1995.
[21] L. M. Sandalio, H. C. Dalurzo, M. Gomez, M. C. Romero-Puertas and L.A. Del Rio. “Cadmium- induced changes in the growth and oxidative metabolism of pea plants,” J. Exper. Bot., vol. 52, pp. 2115-2226, 2001.
[22] M. Havaux, Canaai O. and S. Malkin, “Inhibition of photosynthetic activities under slow water stress measured in vivo by photoacoustic method,” Physiol. Plantarum., vol. 70, pp. 503-510, 1987.
[23] Z. B. Doganlar, K. Demir, H. Basak, and I. Gul, “Effects of salt stress on pigment and total soluble protein contents of three different tomato Cultivars,” Afr. J. Agric. Res., vol. 5, no. 15, pp. 2056-2065, 2010.
[24] L. Jiang, L. Duan, X. Tian, B. Wang and Z. Li, “NaCl salinity stress decreased Bacillus thuringiensis (Bt) protein content of transgenic Bt cotton (Gossypium hirsutum L.) seedlings,” Environ. Exper. Bot., vol. 55, no.3, 315-320, 2005.
[25] V. Chinnusamy, A. Jagendorf and J. K. Zhu, “Understanding and improving salt tolerance in plants,” Crop Sci., vol. 45, pp. 437-448, 2005.
[26] M. Dynowski, G. Schaaf and D. Loque “Plant plasma membrane water channels conduct the signaling molecule H2O2,” J. Biochem. Pharmacol. Res., vol. 414, pp. 53-61, 2008.
[27] K. Sumithra, K. Jutur, B. Dalton-Caramel and A. R. Reddy, “Salinity induced changes in two cultivars of Vigna radiata: response of antioxidative and proline metabolism,” Plant Growth Regul., vol. 50, pp. 11-22, 2006.
[28] F. Queiros, F. Fidalgo, I. Santos and R. Salema, “In vitro selection of salt tolerant cell lines in Solanum tuberosum L.,” Biologia Plantarum., vol. 51, no. 4, pp. 728-734, 2007.
[29] D. R. Gossett, E. P. Millhollon and M. C. Lucas, “Antioxidant response to NaCl stress in salttolerant and salt-sensitive cultivars of cotton,” Crop Sci., vol. 34, pp. 706-714, 1994.
[30] D. H. Lee, Y. S. Kim and C. B. Lee, “The inductive responses of the antioxidant enzymes by salt stress in rice (Oryza sativa L.),” J. Plant Physiol., vol. 158, pp. 737-745, 2001.
[31] J. A. Hernandez, M. A. Ferrer, A. Jimenez, A. R. Barcelo and F. Seville, “Antioxidant systems and O-2 /H2O2 production in the apoplast of pea leaves. Its relation with salt-induced necrotic lesions in minor veins,” Plant Physiol., vol. 127, pp. 817-83, 2001.
[32] Z. L. Wang, H. P. Li, M. Fredricksen, Z. Z. Gong, C. S. Kim and C. Q. Zahng, “Expressed sequence tags from Thellungiella halophila, a new model to study plant salt tolerance,” Plant Sci., vol. 166, pp. 609-616, 2004.