Potential regulation of cleistogamy in pigeonpea through jasmonic acid and bHLH transcription factor interactions

This study provides insights into the molecular and hormonal control of cleistogamy in pigeonpea, focusing on bHLH transcription factors and jasmonic acid pathway. Pigeonpea, an annual diploid (2n = 22) grain legume, holds significant nutritional value in cereal-dominated diets. The chasmogamous flowers of pigeonpea have a typical 9 + 1 diadelphous stamen where flowers open pre-fertilization resulting in cross-pollination. In contrast, a cleistogamous genotype characterized by polyadelphous stamens and flowers that open post-fertilization ensuring seed purity was analyzed for identifying causal pathways. Subsequent analysis focused on a set of transcription factors and their interaction with the hormonal networks associated with cleistogamy. Genes of the Jasmonic acid (JA) signaling pathway have been established to play a significant role in inducing cleistogamy and one of the key regulators of the JA pathway is bHLH (basic helix loop helix). A genome-wide survey identified 176 bHLH genes in the pigeonpea genome. Phylogenetic analysis classified 176 bHLH genes into 21 subfamilies distributed randomly across the genome. Gene ontology, cis-motifs analysis in the upstream region, and protein-protein interaction network implied the involvement of these genes in various biological processes. Expression analysis of key genes of the jasmonic acid pathway which includes MYC2 (Cc_bHLH135) along with its interacting partners TIFY/JAZ in chasmogamous and cleistogamous floral tissues revealed their potential role in flower opening. The results of UHPLC-MS/MS quantitation of Jasmonic acid and its bioactive form JA-Ile align with the expression analysis. The congruence of gene expression and hormone profiling highlights the involvement of the JA pathway in regulating flower opening, implying their potential role in cleistogamy in pigeonpea.