Novel assays to monitor gene expression and protein-protein interactions in rice using the bioluminescent protein, NanoLuc

Luciferases have been widely utilized as sensitive reporters to monitor gene expression and protein-protein interactions. Compared firefly luciferase (Fluc), a recently developed luciferase, Nanoluciferase (NanoLuc or Nluc), has several superior properties such smaller size stronger luminescence activity. We compared the reporter of Nluc Fluc in rice (Oryza sativa). In both plant-based two-hybrid split luc complementation (SLC) assays, activity was detected with higher sensitivity specificity than that Fluc. To apply research involving photoperiodic regulation flowering, we made knock-in plant which coding region inserted in-frame OsMADS15 gene, target florigen Hd3a. Strong response Hd3a, change day length, protoplasts single shoot apical meristem, respectively. Our results indicate assay systems will be powerful tools interaction research.