Phylogenomic analysis of MKKs and MAPKs from 16 legumes and detection of interacting pairs in chickpea divulge MAPK signalling modules

The mitogen-activated protein kinase (MAPK)-mediated phosphorylation cascade is a vital component of plant cellular signalling. Despite this, MAPK signalling cascade is less characterized in crop legumes. To fill this void, we present here a comprehensive phylogeny of MAPK kinases (MKKs) and MAPKs identified from 16 legume species belonging to genistoid (Lupinus angustifolius), dalbergioid (Arachis spp.), phaseoloid (Glycine max, Cajanus cajan, Phaseolus vulgaris, and Vigna spp.), and galegoid (Cicer arietinum, Lotus japonicus, Medicago truncatula, Pisum sativum, Trifolium spp., and Vicia faba) clades. Using the genes of the diploid crop chickpea (C. arietinum), an exhaustive interaction analysis was performed between MKKs and MAPKs by split-ubiquitin based yeast two-hybrid (Y2H). Twenty seven interactions of varying strengths were identified between chickpea MKKs and MAPKs. These interactions were verified in planta by bimolecular fluorescence complementation (BiFC). As a first report in plants, four intra-molecular interactions of weak strength were identified within chickpea MKKs. Additionally; two TEOSINTE-BRANCHED1/CYCLOIDEA/PCF (TCP) transcription factors of class I were identified as novel down-stream interacting partners of seven MAPKs. We propose that this highly reliable MAPK interaction network, presented here for chickpea, can be utilized as a reference for legumes and thus will help in deciphering their role in legume-specific events.