Figure 1

Construction of chimeric orf220 gene, transformation into wild type stem mustard and its sub-localization in mitochondria. Orf220, a candidate cytoplasmic male gene associated with sterility in Brassica juncea, was isolated from cytoplasmic male-sterile stem mustard. A mitochondrial targeted peptide (atp2-1) was isolated from Nicotiana encoding the β subunit of F1-ATPase (Figure 1-A). Figure 1-B shows the transgenic regenerations from proximal portions of hypocotyls of cotyledons in stem mustard. Figure 1-C shows the identification of transgenic stem mustard by checking for the presence of the orf220 gene using PCR and RT-PCR. Figure 1-D shows that the mitochondrial targeted peptide (atp2-1) can guide the ORF220 protein to the mitochondria target, while the ORF220 protein is only found in the nucleus in the absence of the mitochondrial targeted peptide (atp2-1). Mt-RFP was used as the mitochondrially-targeted control.