Cucumis sativus glycine rich protein interacts with cucumber mosaic virus 2b protein

The development of plant stress tolerance is critical in agriculture and horticulture. During viral infection (biotic stress), various plant host factors engage in interactions with viral proteins leading to gene expression dynamics, differential complex formation, signaling, and metabolite production and subsequent tolerance/ resistance or susceptibility. In this study, we found that during cucumber mosaic virus (CMV) infection in Cucumis sativus , glycine-rich protein ( Cs GRP) is associated with the CMV 2b protein. Initially, interaction was identi fied by screening the cucumber cDNA library using yeast two-hybrid assay in which the 2b protein of the virus was used as bait protein. Subsequently, the interaction was further validated using in planta bimolecular fluorescence complementation (BiFC) and in vivo far western blotting (FWB) assays. In BiFC, the bimolecular complex of Cs GRP and 2b was observed as punctate spots in Nicotiana benthamiana epidermal cells. In the localization and colocalization studies, Cs GRP and the bimolecular complex of the Cs GRP-2b protein were found to be localized in the cell wall region. Phylogenetic analysis and domain architecture studies revealed that the interacting Cs GRP belongs to the class II of GRPs, and are characterized by signal peptide, glycinerich, and cysteine-rich regions. After CMV infection, the expression of the CsGRP transcript was signi ficantly upregulated at 7 and 14 days post virus inoculation (dpi). These findings shed light on the molecular connections that establish plant -virus interactions and highlight the potential role of Cs GRP in CMV-induced disease development. (c) 2024 Published by Elsevier B.V. on behalf of SAAB.