Populus trichocarpa Coproporphyrinogen III oxidase (PtCPO) localizes in sub-chloroplastic structures

<p>The open reading frame (ORF) of PtCPO (see sequence below) was amplified by PCR from the hybrid poplar Beaupré (P. trichocarpa x P. deltoides) leaf cDNAs. The ORF was fused with the N-terminus of a green fluorescent protein (GFP) into the pICH86988 in order to create an in-frame PtCPO-GFP protein fusion, under the control of a 35S-Ω promoter. The final vector was inserted into Agrobacterium tumefaciens (GV3101 strain). The PtCPO-GFP protein fusion was transiently expressed in N. benthamiana leaf cells using A. tumefaciens leaf infiltration. Two days after infiltration, pavement cells were observed by live-cell imaging with a laser-scanning confocal microscope. Chlorophyll and GFP were excited at 488 nm, and their fluorescent signals were collected between 505-525 nm and 680-700 nm, respectively. Images present a single optical section of 0.8 µm. White arrowheads indicate discrete areas (diameter: 0.1 to 0.3 µm) within the chloroplasts. Scale bar: 5 µm. >PtCPO (98.7 % amino acid identity with Potri.011G023900.1, Phytozome portal http://www.phytozome.net/genePage.php?search=1&detail=1&crown&method=0&searchText=transcriptid%3A27003187/)</p> <p> </p>