Next generation sequencing for understading the role of nitrate reductase in remodeling intermediate metabolism in Phaeodactylum tricornutum. Phaeodactylum tricornutum

We have done next generation sequencing of optically thin, exponentialy growing WT Phaeodactylum tricornutum and our ~50% nitrate reductase knock-down cultures. Culture were grown with and without nitrogen source to improve our undestanding of the pathways regulation under conditions that promote lipid accumulation (N-starvation). Overall design: 3 biologically independent exponentially growing culture of the WT and the NR21 knock-down were pelleted and washed several times with N-free media. Each culture was devided to 2 replicates with initial cell concentration of 2.5X105 cells/mL, one with NaNO3 as nitrogen source and the other without any nitrogen source. The cuture were bubbled for 48 hours and sampled for transcriptome together with other physiological parameters and lipid analysis after reachign exponential growth.