Abstract
A powerful approach for determining the biological functions of genes in an organism is to produce mutants with altered phenotypes and physiological responses. Various approaches for mutagenesis involving chemical, irradiation, and insertional methods have been developed; each has advantages and disadvantages for the study of gene function. In this post-genomic era, the use of reverse genetic approaches to understanding the role of genes in growth and development has become widespread. With development of new techniques such as targeting induced local lesions in genomes (TILLING), ethyl methanesulfonate (EMS) mutagenesis can be used for both forward and reverse genetic studies. Generation of diverse mutant alleles in the same gene provides critical tools to understand the role of these genes in the function of the organism. Here we describe the general method of EMS mutagenesis for the molecular genetic model plant Arabidopsis thaliana.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Greene, E. A., Codomo, C. A., Taylor, N. E., et al. (2003) Spectrum of chemically induced mutations from a large-scale reverse-genetic screen in Arabidopsis. Genetics 164, 731–740.
Hoffmann, G. R. (1980) Genetic effects of dimethyl sulfate, diethyl sulfate, and related compounds. Mut. Res. 75, 63–129.
Krieg, D. R. (1963) Ethyl methanesulfonate-induced reversion of bacteriophage T4rII mutants. Genetics 48, 561–580.
Kovalchuk, I., Kovalchuk, O., and Hohn, B. (2000) Genome-wide variation of the somatic mutation frequency in transgenic plants. EMBO J. 19, 4431–4438.
McCallum, C. M., Comai, L., Greene, E. A., and Henikoff, S. (2000) Targeted screening for induced mutations. Nat. Biotech. 18, 455–457.
Lee, S. Y., Cheong, J. I., and Kim, T. S. (2003) Production of doubled haploids through anther culture of M1 rice plants derived from mutagenized fertilized egg cells. Plant Cell Rep. 22, 218–223.
Jander, G., Baerson, S. R., Hudak, J. A., Gonzalez, K. A., Gruys, K., and Last, R. L. (2003) Ethyl methanesulfonate saturation mutagenesis in Arabidopsis to determine frequency of herbicide resistance. Plant Physiol. 131, 139–146.
Grini, P. E., Schnittger, A., Schwarz, H., Zimmermann, I., Schwab, B., Jürgens, G., and Hülskamp, M. (1999) Isolation of ethyl methanesulfonate-induced gametophytic mutants in Arabidopsis thaliana by a segregation distortion assay using the multimarker chromosome I. Genetics 151, 849–863.
Koornneef, M., Dellaert, L. W. M., and van der Veen, J. H. (1982) EMS-and radiation-induced mutation frequencies at individual loci in Arabidopsis thaliana (L.) Heynh. Mut. Res. 93, 109–123.
Zhu, B., Gu, A., Deng, X., Geng, Y., and Lu, Z. (1995) Effects of caffeine or EDTA post-treatment on EMS mutagenesis in soybean. Mut. Res. 334, 157–159.
Colbert, T., Till, B. J., Tompa, R., et al. (2001) High-throughput screening for induced point mutations. Plant Physiol. 126, 480–484.
Henikoff, S. and Comai, L. (2003) Single-nucleotide mutations for plant functional genomics. Annu. Rev. Plant Biol. 54, 375–401.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2006 Humana Press Inc.
About this protocol
Cite this protocol
Kim, Y., Schumaker, K.S., Zhu, JK. (2006). EMS Mutagenesis of Arabidopsis . In: Salinas, J., Sanchez-Serrano, J.J. (eds) Arabidopsis Protocols. Methods in Molecular Biology™, vol 323. Humana Press. https://doi.org/10.1385/1-59745-003-0:101
Download citation
DOI: https://doi.org/10.1385/1-59745-003-0:101
Publisher Name: Humana Press
Print ISBN: 978-1-58829-395-4
Online ISBN: 978-1-59745-003-4
eBook Packages: Springer Protocols