Subcellular Localization

The putative plastid transit peptides of DXS isoforms were predicted with TargetP 1.1. The fragments of three DXS genes amplified with specific primers were inserted into the BamHI site of the pTF486 expression vector, which includes an enhanced green fluorescent protein (EGFP) coding sequence ( Table S1 ). For each recombinant plasmid, approximately 2 μg was introduced into A. thaliana mesophyll protoplasts via polyethylene glycol-mediated transformation (Yoo et al., 2007). The EGFP fluorescence and chlorophyll autofluorescence were observed with the FV1200 confocal laser scanning microscope (Olympus, Tokyo, Japan) at excitation wavelengths of 488 and 561 nm, respectively.