Characterization of drought tolerance of GmDREB2 soybean mutants (Glycine max (L.) Merr) by ethyl methane sulfonate induction

摘要

Mutation induction using the chemical mutagen ethyl methane sulfonate (EMS) was proposed for soybean breeding. Such mutation events result in the exchange of bases of DNA that will cause genotypic and phenotypic changes. Breeding mutations can be used to obtain improved varieties. The DREB2 gene is a subclass of the DREB (dehydration responsive element binding) gene family of transcription factors. It serves to respond to and regulate gene expression during drought stress. The aims of this study were to identify and to characterize the GmDREB2 gene with various treatments of EMS mutation induction. Treatments included combinations of EMS concentration (0.05%, 0.50% and 1.00%) and immersion time (4, 6 and 8 hours). The DNA genomes of these varieties were isolated by using methods from Doyle & Doyle. DNA amplification was performed using polymerase chain reaction with a specific primer designed on the basis of GmDREB2 sequence gene data from NCBI. The DNA resulting from amplification was sequenced using an automatic sequencing machine (BigDye Terminator v3.1). The results of the research showed that the sequences of GmDREB2 in the treatment of 1.00% EMS with 8 hours immersion time showed the greatest change, not only in the sequence of the GmDREB2 gene but also the change in amino acids.