The<i>C</i>_<i>4</i><i>Ppc</i>promoters of many C₄grass species share a common regulatory mechanism for gene expression in the mesophyll cell

Abstract

C₄ photosynthetic plants have evolved from C₃ ancestors and are characterized by differential expression of several hundred genes. Strict compartmentalization of key C₄ enzymes either to mesophyll (M) or bundle sheath cells is considered a crucial step towards the evolution of C₄ photosynthesis. In this study, we demonstrate that the 5'-flanking sequences of the C₄ type phosphoenolpyruvate carboxylase (Ppc) gene from three C₄ grass species could drive M-cell-specific expression of a reporter gene in rice. In addition to that, we identified about 450 bp (upstream of their transcription start site) of the analyzed C₄ Ppc promoters contain all the essential regulatory elements for driving M-cell-specific expression in rice leaves. Importantly, four motifs of conserved nucleotide sequences (CNSs) were also determined, which are essential for the activity of the promoter. A putative interaction between the CNSs and an unknown upstream element(s) is required for driving M-cell-specific expression. This work identifies the evolutionary conservation of C₄ Ppc regulatory mechanisms of multiple closely related C₄ grass species.