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Metal tolerance proteins (MTPs) encompass plant membrane divalent cation transporters to specifically participate in heavy metal stress resistance and mineral acquisition. However, the molecular behaviors and biological functions of this family in  Medicago truncatula  are scarcely known. A total of 12 potential  MTP  candidate genes in the  M. truncatula  genome were successfully identified and analyzed for a phylogenetic relationship, chromosomal distributions, gene structures, docking analysis, gene ontology, and previous gene expression.  M. truncatula MTPs  ( MtMTPs ) were further classified into three major cation diffusion facilitator (CDFs) groups: Mn-CDFs, Zn-CDFs, and Fe/Zn-CDFs. The structural analysis of  MtMTPs  displayed high gene similarity within the same group where all of them have cation_efflux domain or ZT_dimer.  Cis -acting element analysis suggested that various abiotic stresses and phytohormones could induce the most  MtMTP  gene transcripts. Among all  MTPs , PF16916 is the specific domain, whereas GLY, ILE, LEU, MET, ALA, SER, THR, VAL, ASN, and PHE amino acids were predicted to be the binding residues in the ligand-binding site of all these proteins. RNA-seq and gene ontology analysis revealed the significant role of  MTP  genes in the growth and development of  M. truncatula .  MtMTP  genes displayed differential responses in plant leaves, stems, and roots under five divalent heavy metals (Cd 2+ , Co 2+ , Mn 2+ , Zn 2+ , and Fe 2+ ). Ten, seven, and nine  MtMTPs  responded to at least one metal ion treatment in the leaves, stems, and roots, respectively. Additionally,  MtMTP1.1 ,  MtMTP1.2 , and  MtMTP4  exhibited the highest expression responses in most heavy metal treatments. Our results presented a standpoint on the evolution of  MTPs  in  M. truncatula . Overall, our study provides a novel insight into the evolution of the  MTP  gene family in  M. truncatula  and paves the way for additional functional characterization of this gene family.

frontiers in genetics

Genome-Wide Identification and Expression Analysis of Metal Tolerance Protein Gene Family in Medicago truncatula Under a Broad Range of Heavy Metal Stress