Submitted to: Biologia Plantarum
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 6/3/2003
Publication Date: 3/1/2004
Citation: Hofmann, N.E., Raja, R., Nelson, R.L., Korban, S.S. 2004. Mutagenesis of embryogenic cultures of soybean and detecting polymorphisms using rapd markers. Biologia Plantarum. 48: 173-177. Interpretive Summary: Mutagenesis is means of creating genetic diversity that does not exist or has not been found in existing germplasm. Treating somatic embryos, embryos derived from culturing portions of immature soybean seeds, with mutagenic agents can be an efficient method of creating mutations because they are easier to regenerate into whole plants than cell cultures and easier to handle in large numbers than seeds. Our research demonstrated that RAPDs, a type of DNA marker, can be a useful tool to assess the occurrence of mutations in treated embryo cultures. This information will useful to geneticist and physiologists who work with cell and embryo cultures.
Technical Abstract: Embryogenic suspension cultures of soybean (Glycine max L. cv. Iroquois) were subjected to mutagenesis using varying concentrations (1, 3, 10, and 30 mM) of ethyl methanesulfonate (EMS). Depending on the concentration of EMS used, the mean survival rate of embryogenic cultures decreased from 74% (1 mM EMS) to 43% after 30 mM EMS treatment. Random amplified polymorphic DNA (RAPD) analysis was used to determine whether induction of genetic variability in embryogenic cultures in response to the different EMS treatments may result in identification of polymorphic markers. Two of 35 'core' primers tested revealed polymorphisms. One of the primers, OPO-01/1150, revealed polymorphism in tissue treated with 10 mM EMS, while the other primer, OPO-05/1200, revealed polymorphism in tissue treated with either 1 or 30 mM EMS. These results suggest that RAPD markers are useful in detecting mutations in embryogenic cultures of soybean.