Genome-wide identification and expression analysis of Raf-like kinase gene family in pepper (Capsicum annuum L.)

As highly conserved signaling pathway modules, mitogen-activated protein kinase (MAPK) cascades play vital roles in a diverse range of stress and hormonal responses in plants. Among the established components of MAPK cascades, Raf-like MAPK kinase kinases (MAPKKKs) are associated with abscisic acid (ABA) signaling and osmotic stress responses. However, despite the availability of a pepper reference genome, few of the Raf-like kinases in pepper plants have been functionally characterized. In this study, we isolated 47 putative Raf-like kinase genes from the pepper genome based on in silico analysis, which were classified into two major categories, namely, groups B and C (further sub-grouped into B1–B4 and C1–C7, respectively) and named sequentially as CaRaf1 to CaRaf47. Subcellular localization prediction analysis revealed that most of the group B CaRaf-like kinases are probably nuclear-localized, whereas a majority of group C members targeted into the cytoplasm. Transcriptional regulation of the 47 CaRaf genes in response to treatment with ABA, drought, NaCl, and mannitol was quantitatively analyzed by reverse-transcription PCR analysis. This revealed a significant induction of subgroup B3, C2, C3, and C5 members, indicating that these genes may be functionally associated with the response to osmotic stress, mediated via both ABA-dependent and -independent pathways. The findings of this study can accordingly serve as a basis for the identification of CaRaf genes associated with the regulation of ABA signaling and osmotic stress response and thus contribute to enhancing our understanding of the biological functions of CaRaf kinases in the responses of plants to different abiotic stresses.