Transcriptome analysis of Nicotiana benthamiana leaves at 96 hours post-infiltration with a plastid-localized bacterial phytoene synthase (p)crtB
Identifiers: SRA: SRP238752
N. benthamiana leaves were agroinfiltrated with 35S:pcrtB (or 35S:GFP as control) and sampled at 96 hours post infiltration. The second or third youngest leaves of 4 to 6 week old N. benthamiana plants were infiltrated with A. tumefaciens strain GV3101 carrying the plasmids of interest. Gene silencing was prevented by co agroinfiltration with a strain carrying the helper component protease HcPro of the watermelon mosaic virus WMV in plasmid HcProWMV pGWB702. Infiltration cultures were grown on LB medium at 28 degrees and used at optical density at 600 nm of 0.5. Total RNA was extracted from leaves with the Maxwell 16 LEV Plant RNA Kit. Indexed libraries were prepared from 1 ug of purified RNA with Illumina TruSeq Stranded mRNA Sample Prep Kit according to the manufacturer instructions. Libraries were quantified using the TapeStation 4200 and pooled such that each index tagged sample was present in equimolar amounts, with final concentration of the pooled samples of 2 nM. The pooled samples were subjected to cluster generation and sequencing using a Illumina NextSeq 500 System in a 2x75 paired end format at a final concentration of 1.8 pmol. pcrtB sections were compared to green GFP controls.
External Link: Synthetic biogenesis of chromoplasts from leaf chloroplasts