Meristem defective protein (MDF) plays its role in proper development of meristem patterns in Arabidopsis through regulation of auxins homeostasis. It also enables plants to survive in stress. Present study was designed to characterize MDF protein in A. thaliana to get an insight into its molecular aspects. Sequence of protein was retrieved from uniprot database and subjected to CELLO, SOPMA, SWISSMODEL, MEME server and SRING tools. The protein was also docked with four proteins i. e. SR4, RSZ33, PLT1, 2 and 3. The protein was found to localize in nucleus with secondary structure comprising of alpha helix (47.20%), extended strand (6.34%), beta turn (5.37%) and random coil (41.10%). MDF was analyzed as a conserved protein with monomeric structure. Proteins observed as interacting partners of MDF included STA1, T13D8.9, LSM5, LSM2, LSM4 and LSM8. Docking analysis revealed highest and lowest affinities of MDF binding with RS4, RZS33, PLT1, PLT2 and PLT3 in case of FGRTLTPKEAFRLLSHKFHG and IQGQTTHTFEDLNSSAKVSSDYFSQ conserved motifs, respectively. Features of MDF protein explored in this investigation can be exploited for engineering and production of plants with efficient root meristems development.