A lipid droplet-associated protein involved in lipid droplet biogenesis and triacylglycerol accumulation in the oleaginous microalga Phaeodactylum tricornutum

Lipid droplets (LDs) are lipid monolayer-enclosed organelles comprising a lipid core and surface associatedproteins. However, the protein components and their regulatory functions in LDs have remained largely unknown in oleaginous diatoms. In this study, we identified a gene encoding lipid droplet (LD)-associated protein (PtLDP1) in Phaeodactylum tricornutum and examined its function. The PtLDP1 showed homology to the diatomoleosome-associated protein 1 (DOAP1) from Fistulifera. Overexpression of the PtLDP1 gene elevated lipid content, enlarged LD size and increased relative expression levels of key genes involved in triacylglycerol (TAG) and fatty acid biosynthesis. In contrast, knockdown of PtLDP1 by RNAi decreased lipid and TAG content, and subsequently reduced LD size. In addition, LDs were isolated from P. tricornutum cells and the proteome of LDs was identified by mass spectrometry. We found that PtLDP1 was a significant protein in the LD proteome. Importantly, labeling of enhanced yellow fluorescent protein (EYFP) confirmed that the PtLDP1 was localized to the LDs. Altogether, our data suggest that the PtLDP1 could be an important LD-associated protein contributing to regulation of TAG synthesis and lipogenesis. The findings will provide new targets for genetic improvement of oleaginous microalgae.