Biological and genomic analysis of a symbiotic nitrogen fixation defective mutant in Medicago truncatula

Medicago truncatula has been selected as one of the model legume species for gene functional studies. To elucidate the functions of the very large number of genes present in plant genomes, genetic mutant resources are very useful and necessary tools. Fast Neutron (FN) mutagenesis is effective in inducing deletion mutations in genomes of diverse species. Through this method, we have generated a large mutant resource in M. truncatula. This mutant resources have been used to screen for different mutant using a forward genetics methods. We have isolated and identified a large amount of symbiotic nitrogen fixation (SNF) deficiency mutants. Here, we describe the detail procedures that are being used to characterize symbiotic mutants in M. truncatula. In recent years, whole genome sequencing has been used to speed up and scale up the deletion identification in the mutant. Using this method, we have successfully isolated a SNF defective mutant FN007 and identified that it has a large segment deletion on chromosome 3. The causal deletion in the mutant was confirmed by tail PCR amplication and sequencing. Our results illustrate the utility of whole genome sequencing analysis in the characterization of FN induced deletion mutants for gene discovery and functional studies in the M. truncatula. It is expected to improve our understanding of molecular mechanisms underlying symbiotic nitrogen fixation in legume plants to a great extent.

Automated summary

Medicago truncatula has been chosen as a model legume species for gene functional studies. Fast Neutron (FN) mutagenesis has been used to generate a large mutant resource in M. truncatula, which has been utilized to screen for symbiotic nitrogen fixation deficiency mutants. Whole genome sequencing has been employed to identify a SNF defective mutant FN007, which has a large segment deletion on chromosome 3. These findings demonstrate the usefulness of whole genome sequencing in characterizing FN induced deletion mutants and advancing our understanding of symbiotic nitrogen fixation in legume plants.