Mitogen-Activated Protein Kinase Kinase Kinases (MAPKKKs/MAP3Ks) are widespread signal transduction module of MAPK cascades and show significant role in growth, development and defense/tolerance responses to environmental stresses. However, MAPKKKs are not yet identified and characterized in the sequenced Cicer arietinum genome and their control on plant defense response against herbivory is still poorly understood. Therefore, a comprehensive investigation of chickpea genome was performed, which identified 52 MAPKKKs (CaMAPKKKs). The phylogenetic study of MAPKKKs from chickpea, Arabidopsis and rice, categorized them into three subclasses, namely MEKK, RAF and ZIK. Presence of respective conserved sequences in each CaMAPKKKs strongly supported their identity as members of the three subfamilies. Physical mapping of these genes revealed their wider distribution on all the eight chromosomes. The three-dimensional structure prediction and potential active sites identification of selected MAPKKK proteins revealed the presence of phosphotransferase domain, which is required for its catalytic activity and participation in the MAPK cascade. The relative expression level of 29 CaMAPKKKs transcripts exhibited differential expression in diverse tissue types, but others showed no/very low expression. Further, in order to figure out their involvement in plant defense response, the expression patterns of CaMAPKKK genes was evaluated during Helicoverpa armigera –feeding by qPCR, which revealed their differential expression during herbivory. This study on identification and characterization of CaMAP3K gene family provided insights into evolution and divergence of CaMAPKKKs and their significance in plant defense against herbivores, which can be further utilized to decipher MAPK pathway involved in plant defense in chickpea and also for pest management.