Determination of LD50 of Ethidium Bromide for Induction of Mutation in Marigolds

##plugins.themes.bootstrap3.article.main##

Pornthep Keadtidumrongkul Nawannaporn Chirarat Sophana Somran

Abstract

     Mutation is an effective way to increase genetic variability in plant, which can be done in two ways: physical mutation and chemically induced mutation using a chemical mutagen. Ethidium bromide (EtBr) is the most efficient chemical mutagen,  inducing mutation in the mitochondria, chloroplasts and nucleus DNA. To increase genetic variance in marigolds, the appropriate concentration level and soaking periods of EtBr must be identified, which was the purpose of this study. Marigold seeds were soaked with 0, 250, 500, 750 and 1000 mg/l of EtBr for 24, 48 and 72 hours. The results showed that the most appropriate concentration of EtBr for inducing mutation in marigolds (LD50) was 623.81 mg/l for 48 hours. However, increasing the concentration level of EtBr decreased the growth of the marigolds, as measured by the height of the tree, and, further, EtBr markedly reduced the survival percentage, especially at high concentration levels.


Keywords: mutagen, genetic variance, lethal dose

References

Anbarasn K., Sivalingam, D., Rajendran, R., Anbazhagan, M., & Chidambaram, AI. A. (2013). Studies on the mutagenic effect of EMS on seed germination and seedling characters of Sesame (Sesamum indicum L.) Var. T MV3. International Journal of Biological Sciences, 3, 68-70.
Burton, G. W., & Hanna, W. W. (1976). Ethidium Bromide Induced Cytoplasmic Male Sterility in Pearl Millet1. Crop Science, 16, 731-732. https://doi:10.2135/cropsci1976.0011183X0016000500 35x
Chopra, V. L. (2005). Mutagenesis: Investigating the process and processing the outcome for cropimprovement. Current Science, 89, 353-359.
Deepak, P., Kadambari, G., & Abha, A. (2011). Can we predict mutagen-induced damage in plant systems mathematically? Insights from zygotic embryo and haploid mutagenesis in Indian mustard (Brasica juncea). Botanica Serbica, 35(2), 137-143.
Fukunaga, M., Cox, B. A., Sprecken, R. S., & Yielding, L. W. (1984). Production of frameshift mutations in Salmonella by phenanthridinium derivatives: enzymatic activation and photoaffinity labeling. Mutation Research/Fundamental and Molecular Mechanisms of Mutagenesis, 127(1), 31-37. https://doi.org/ 10.1016/0027-5107(84)90137-4
Goldring, E. S., Grossman, L. I., Krupnick, D., Cryer, D. R., & Marmur, J. (1970). The petite mutation in Yeast. Journal of Molecular Biology, 52(2), 323-335.
Hsu, T. C. (1968). Action of Chemicals on Dividing Cells. New Jersey: Prentice-Hall, Inc.
Kharkwal, M. C., & Shu, Q. Y. (2009) The Role of Induced Mutations in World Food Secuity. Retrieved from https://mvd.iaea.org/PDF/IPM200901.pdf
Levy, A., & Ashiri, A. (1975). Ethidium bromide—An efficient mutagen in higher plants. Mutation Research/ Fundamental and Molecular, 28(3), 397-404.
Levy, A., Ashri, A., & Rubin, B. (1978). Ethidium bromide uptake by peanut seeds and its relationship to varietal sensitivity and mutagenic efficiency. Environmental and Experimental Botany, 19(1), 49-55.
McCann, J., & Ames, B. N. (1976). Detection of carcinogens as mutagens in the Salmonella/microsome test: assay of 300 chemicals: discussion. Proceedings of the National Academy of Sciences of the United States of America, 73(3), 950-954.
Miki, T., & Hardt, W.-D. (2013). Outer membrane permeabilization is an essential step in the killing of gram-negative Bacteria by the Lectin RegIIIβ. Plos One, 8(7), e69901. https://doi:10.1371/journal. pone.0069901
Ohta, T., Tokishita, S.-i., & Yamagata, H. (2001). Ethidium bromide and SYBR Green I enhance the genotoxicity of UV-irradiation and chemical mutagens in E. Coli. Mutation. Mutation Research/Genetic Toxicology and Environmental Mutagenesis, 492, 91-97.
Roslim, D. I., Herman, & Fiatin, I. (2015). Lethal dose 50 (LD50) of mungbean (Vigna radiata L. Wilczek) cultivar kampar. SABRAO Journal of Breeding & Genetics, 47(4), 510-516.
Singh, B. D. (2000). Mutations in Crop Improvement. In: Plant Breeding: Principles and Methods. New Delhi: Kalyani Publishers.
Waring, M. (1971). Binding of drugs to supercoiled circular DNA: Evidence for and against intercalation. In F. E. Hahn (Ed.), Proceedings of the Research Symposium on Complexes of Biologically Active Substances with Nucleic Acids and Their Modes of Action: Held at the Walter Reed Army Institute of Research Washington, 16–19 March 1970 (pp. 216-231). Berlin, Heidelberg: Springer Berlin Heidelberg.
Whittaker, P. A., Hammond, R. C., & Luha, A. A. (1972). Mechanism of Mitochondrial Mutation in Yeast. Nature New Biology, 238, 266. https://doi:10.1038/newbio238266a0

Section
Research Articles

##plugins.themes.bootstrap3.article.details##

How to Cite
KEADTIDUMRONGKUL, Pornthep; CHIRARAT, Nawannaporn; SOMRAN, Sophana. Determination of LD50 of Ethidium Bromide for Induction of Mutation in Marigolds. Naresuan University Journal: Science and Technology (NUJST), [S.l.], v. 26, n. 4, p. 80-88, nov. 2018. ISSN 2539-553X. Available at: <http://www.journal.nu.ac.th/NUJST/article/view/Vol-26-No-4-2018-80-88>. Date accessed: 24 july 2019. doi: https://doi.org/10.14456/nujst.2018.24.