Effect of Chemical Mutagen on Some Biochemical Properties of Stevia rebaudiana Bertoni

Document Type : Research Article

Authors

1 Department of Agriculture, Faculty of Biology, Sana Institute of Higher Education, Sari, Iran

2 Department of Biology, Faculty of Agriculture, Islamic Azad University, Damghan, Iran

3 Department of Agronomy & Plant Breeding, Faculty of Genetics and Agricultural Biotechnology Institute of Tabarestan, Sari University of Agricultural Sciences and Natural Resources, Sari, Iran

Abstract

Induced mutagenesis causes an increase variation of some products with limited genetic resources. In this study, the antioxidant enzymes activity, biochemical properties and glycoside content of Stevia were assessed by EMS (chemical mutagen) based on two experiments as factorial with completely randomized design with three replications. The results of the first experiment showed that some properties of regenerated calli were influenced by different concentrations of EMS, different times of exposure and interactions of these two factors. Therefore, more EMS concentration and its duration of exposure indicated fewer regeneration rates and indices of the calcified masses. According to the second experiment, our data showed that EMS mutant activity was related to enzymatic activity and some biochemical properties of regenerated plants at a 1% significance level. Among the mutants, M10, M6, and M19 had the highest enzyme activity and M3 and M10 mutants with + 41.4 and +26.12% revealed the higher percentage of incremental changes in proline accumulation than the control sample. In addition, the highest amount of total protein was allocated to M16, M14 and M3 mutants (values of 0.69, 0.67 and 0.58 mg/g tissue texture, respectively) over the control sample (0.27 mg/ gram of tissue) and M8 mutant had the highest percentage of changes in the amount of stevioside (87.3%) and rebaudioside A (58.3%), respectively. Overall, significant changes were observed in the antioxidant activity and biochemical properties and the amount of sugary glycosides of regenerated plants that could be used to create plants with higher quality traits.

Keywords


Akhtar N. 2014. Effect of physical and chemical mutagens on morphological behavior of tomato (Solanum Lycopersicum) CV. “Rio Grande” under heat stress conditions. Plant Breed Seed Sci 70: 69-79.
Al-Rumaih MM, Al-Rumaih MM. 2008. Influence of ionizing radiation on antioxidant enzymes in three species of Trigonella. Am J Environ Sci 4: 151-156.
Arulbalachandran D, Mullainathan L. 2009. Chlorophyll and morphological mutants of black gram (Vigna mungo (L.) Hepper) derived by gamma rays and EMS. J Phytol 1(4):236-241.
Bates LS, Waldren RP, Teare ID. 1973. Rapid determination of free proline for water-stress studies. Plant Soil 39: 205-207.
Bohnert HJ, Nelson DE, Jensen RG. 1995. Adaptations to environmental stresses. Plant Cell 7(7): 1099-1111.
Borzouei A, Kafi M, Sayahi R, Rabiei E, Amin PS. 2013. Biochemical response of two wheat cultivars (Triticum aestivum L.) to gamma radiation. Pak J Bot 45(2):473-477.
Bradford MM. 1976. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254.
Chang A, Lim MH, Lee SW, Robb EJ, Nazar RN. 2008. Tomato phenylalanine ammonia-lyase gene family, highly redundant but strongly underutilized. J Biol Chem 283: 33591-33601.
de Camargo, AC, Vieira TMFDS, Regitano-D’Arce MAB, de Alencar SM, Calori-Domingues MA, Canniatti-Brazaca SG. 2012. Gamma radiation induced oxidation and tocopherols decrease in in-shell, peeled and blanched peanuts. Int J Mol Sci 13: 2827-2845.
El Sherif F, Khattab S, Ghoname E, Salem N, Radwan K. 2011. Effect of gamma irradiation on enhancement of some economic traits and molecular changes in Hibiscus sabdariffa L. Life Sci J 8: 220-229.
Fang J, Nakamura H, Maeda H. 2011. The EPR effect: unique features of tumor blood vessels for drug delivery, factors involved, and limitations and augmentation of the effect. Adv Drug Deliv Rev 63: 136-151.
Fang JY, Traore S. 2011. In vitro mutation induction of Saintpaulia using ethyl methanesulfonate. HortScience 46: 981-984.
Harrison K, Were LM. 2007. Effect of gamma irradiation on total phenolic content yield and antioxidant capacity of almond skin extracts. Food Chem 102: 932-937.
In BC, Motomura S, Inamoto K, Doi M, Mori G. 2007. Multivariate Analysis of Relations between pre harvest environmental factors, postharvest morphological and physiological factors, and vase life of cut ‘Asami Red’ roses. J Jpn Soc Hortic Sci 76: 66-72.
Jain SM, Spencer MM. 2006. Biotechnology and mutagenesis in improving ornamental plants in floriculture and ornamental biotechnology. Advances and Tropical Issues, In: Teixeira da Silva JA, eds. London: Global Science Books Ltd, 1749-2036.
Jang E, Lee H, Kim J, Lee S. 2005. Effects of low dose γ-ray irradiation on antioxidant activity of seeds and seedling growth in Raphanus sativus L. Korean J Hortic Sci Technol 23: 245-249.
Khalil SA, Ahmad N, Zamir R. 2015. Gamma radiation induced variation in growth characteristics and production of bioactive compounds during callogenesis in Stevia rebaudiana (Bert.). New Negatives Plant Sci 1: 1-5.
Khalil SA, Zamir R, Ahmad N. 2014. Effect of different propagation techniques and gamma irradiation on major steviol glycoside’s content in Stevia rebaudiana. J Animal Plant Sci 24: 1743-51.
Khan IA, Dahot MU, Khatri A. 2007. Study of genetic variability in sugarcane induced through mutation breeding. Pak J Bot 39: 1489-1501.
Kiong ALP, Lai AG, Hussein S, Harun AR. 2008. Physiological responses of Orthosiphon stamineus plantlets to gamma irradiation. Am-Eurasian J Sustain Agric 2: 135-149.
Lee SY, Cheong JI, Kim TS. 2003. Production of doubled haploids through anther culture of M1 rice plants derived from mutagenized fertilized egg cells. Plant Cell Rep 22: 218-223.
Lemus-Mondaca R, Vega-Gálvez A, Zura-Bravo L, Ah-Hen K. 2012. Stevia rebaudiana Bertoni, source of a high-potency natural sweetener: A comprehensive review on the biochemical, nutritional and functional aspects. Food Chem 132: 1121-1132.
Luck H. 1974. Peroxidase, p. 895-897. In H-U. Bergmeyer (ed.), Methods of enzymatic analysis. Academic Press, Inc., New York.
Ma L, Zhou E, Gao L, Mao X, Zhou R, Jia J. 2008. Isolation, expression analysis and chromosomal location of P5CR gene in common wheat (Triticum aestivum L.). South African J Bot 74: 705-712.
Maslobrod SN, Korletyanu LB, Ganya AI. 2010. Influence of millimetric radiation on the viability of plants: Changing the metabolism of seeds at the factor’s influence on dry seeds. Surf Eng Appl Electrochem 46: 477-488.
Rahi P, Pathania V, Gulati A, Singh B, Bhanwra RK, Tewari R. 2010. Stimulatory effect of phosphate-solubilizing bacteria on plant growth, stevioside and rebaudioside-A contents of Stevia rebaudiana Bertoni. Appl Soil Ecol 46: 222-229.
Raymond J, Rakariyatham N, Azanza JL. 1993. Purification and some properties of polyphenoloxidase from sunflower seeds. Phytochem 34: 927-931.
Sánchez-Rodríguez E, Rubio-Wilhelmi M, Cervilla LM, Blasco B, Rios JJ, Rosales MA, Ruiz JM. 2010. Genotypic differences in some physiological parameters symptomatic for oxidative stress under moderate drought in tomato plants. Plant Sci 178: 30-40.
Sgherri CLM, Maffei M, Navari-Izzo F. 2000. Antioxidative enzymes in wheat subjected to increasing water deficit and rewatering. J Plant Physiol 157: 273-279.
Shevyakova NI, Bakulina EA, Kuznetsov VV. 2009. Proline antioxidant role in the common ice plant subjected to salinity and paraquat treatment inducing oxidative stress. Russian J Plant Physiol 56: 663-669.
Van Harten AM. 1998. Mutation breeding: theory and practical applications. Cambridge University Press.
Wang JW, Zheng LP, Wu JY, Tan RX. 2006. Involvement of nitric oxide in oxidative burst, phenylalanine ammonia-lyase activation and taxol production induced by low-energy ultrasound in Taxus yunnanensis cell suspension cultures. Nitric Oxide 15: 351-358.
Wattoo JI, Aslam K, Shah SM, Shabir G, Sabar M, Naveed SA, Arif, M. 2012. Ethyle methane sulphonate (EMS) induced mutagenic attempts to create genetic variability in Basmati rice. J Plant Breed Crop Sci 4: 101-105.
Waugh R, Leader DJ, McCallum N, Caldwell D. 2006. Harvesting the potential of induced biological diversity. Trends Plant Sci 11: 71-79.
Wi SG, Chung BY, Kim JS, Kim JH, Baek MH, Lee JW, Kim YS. 2007. Effects of gamma irradiation on morphological changes and biological responses in plants. Micron 38: 553-564.