Lipid turnover between membrane lipids and neutral lipids via inhibition of diacylglyceryl N,N,N-trimethylhomoserine synthesis in Chlamydomonas reinhardtii

Chlamydomonas reinhardtii of the microalgal model species lacks phosphatidylcholine (PC), and PC is replaced by diacylglyceryl N,N,N-trimethylhomoserine (DGTS). DGTS is a betaine lipid that is placed in the endoplasmic reticulum (ER) and is synthesized by a single gene, BTA1. In this study, we aimed to ascertain the turnover between membrane and neutral lipids via BTA1 knockdown transformants. Transgenic lines CrBta-hm13 and CrBta-hm31 were 80% and 60% downregulated in BTA1 gene expression levels, respectively. Both transformants had half the amount of DGTS, which coincided with decreased monogalactosyldiacylglycerol (MGDG), which was increased approximately threefold in neutral lipids. While the reduction of DGTS was shown to arise from inhibition of DGTS synthesis, decreased MGDG was affected by internal stress, such as ER stress, which was induced to have a decreased amount of DGTS in the ER. However, galactolipids, except for MGDG, and phospholipids in the transformants were maintained at similar levels. In the transformants, the molar proportion of C16:4 and C18:3(9,12,15), which are the major fatty acids of MGDG, was significantly increased in triacylglycerol (TAG) because of MGDG degradation. Thus, lipid turnover arising from the downregulation of DGTS and induction of ER stress caused a decrease in DGTS and MGDG, which generated a synergy effect on the accumulation of TAG. This study implies that genetic modification of a membrane lipid synthesis pathway could not only be a suitable approach to target accumulation of TAG, but could also suggest a mechanism for the lipid turnover between membrane lipids and neutral lipids.