Gene expression and characterization of an antimicrobial peptide from Medicago sativa "Sazova" cultivar

In recent years, the discovery of new antimicrobial agents has become necessary because of the increase in antibiotic resistance, the development of herbicides and fungicides resistance. Among the antimicrobial agents, antimicrobial peptides (AMPs) stand out due to their stable structure. In this study, the aim was to identify a thermostable AMP from the seeds of M. sativa "Sazova" cultivar and to analyze gene expression during germination. Antimicrobial tests were performed for the seed peptides after heat treatment (85 °C for 10 min), revealing antimicrobial effects against S. aureus, E. coli, and C. albicans. Subsequently, the peptide band corresponding to the inhibition zone was identified as M. sativa Defensin 2.1 (MsDef2.1, MW: 5.2048 kDa). The gene expression analysis of MsDef2.1 in Sazova cultivar showed that the gene was expressed different plant organs, and the expression was decreased over time. As a result of the gene analysis of two cultivars (Sazova and LegenDairy) it was found that there are 5 base differences in the coding sequence and 3 amino acid differences between the sequences of MsDef2.1 isoforms from the LegenDairy and Sazova cultivars. The physiochemical properties, secondary, and tertiary structure of the Sazova Defensin 2.1 were predicted by using bioinformatic tools. Due to the amino acid substitutions in γ-core structures, the antimicrobial activity of the isoforms is expected to differ from each other. These findings demonstrated that the defensin MsDef2.1 can differ in M. sativa cultivars in respect of the gene and amino acid sequences and has a potential for future applications.