Analysis of the root mRNA interactome from canola and rice: Crop species that span the eudicot-monocot boundary

The advent of RNA interactome capture (RIC) has been important in characterizing the mRNA-binding proteomes (mRBPomes) of several eukaryotic taxa. To date, published plant poly(A)⁺ RIC studies have been restricted to Arabidopsis thaliana and specific to seedlings, suspension cell cultures, mesophyll protoplasts, leaves and embryos. The focus of this study was to expand RIC to root tissue in two crop species, the oilseed eudicot Brassica napus (canola) and the cereal monocot Oryza sativa (rice). The optimization and application of root RIC in these species resulted in the identification of 499 proteins and 334 proteins comprising the root mRBPomes of canola and rice, respectively, with 182 shared orthologous proteins between these two species. In both mRBPomes, approximately 80 % of captured proteins were linked to RNA biology, with RRM-containing proteins and ribosomal proteins among the most overrepresented protein groups. Consistent with trends observed in other RIC studies, novel RNA-binding proteins were captured that lacked known RNA-binding domains and included numerous metabolic enzymes. The root mRBPomes from canola and rice shared a high degree of similarity at the compositional level, as shown by a comparative analysis of orthologs predicted for captured proteins to the published Arabidopsis RIC-derived mRBPomes, as well as our Arabidopsis root mRBPome data presented here. This analysis also revealed that 46 proteins in the canola and rice root mRBPomes were unique when orthologs were compared to the published Arabidopsis RBPomes, including those identified recently using phase separation approach that identified proteins bound to all RNA types. The results from this research expands the plant mRBPome into root tissue using two crop species that span the eudicot-monocot clade boundary, and provides fundamental knowledge on RNA-binding protein function in post-transcriptional control of gene expression in crop species for possible future development of beneficial traits.