Overexpression of NSP2 in Parasponia andersonii

NSP2 over-expression was shown to regulate a range of signalling pathways in barley and M. truncatula (Li et al. 2022), in this study we questioned the conservation of NSP2 regulation of signalling pathways in P. andersonii. To investigate that transgenic lines over-expressing P. andersonii NSP2 (PanNSP2) with gradient levels of gene expression were created, following three strategies mutating the putative site of microRNA targeting PanNSP2 messenger RNA (mRNA), inserting into PanNSP2 coding sequence (CDS) an intron known to enhance gene expression, and driving PanNSP2 CDS by a constitutive promoter.