RESUMEN
Lysophosphatidic acid acyltransferase1 (LPAT1) catalyzes the second step of de novo glycerolipid biosynthesis in chloroplasts. However, the embryonic-lethal phenotype of the knockout mutant suggested an unknown role for LPAT1 in non-photosynthetic reproductive organs. Reciprocal genetic crossing of the lpat1-1 heterozygous line suggested a female gametophytic defect of the lpat1-1 knockout mutant. By suppressing LPAT1 specifically during seed development, we showed that LPAT1 suppression affected silique growth and seed production. Glycerolipid analysis of the LPAT1 knockdown lines revealed a pronounced decrease of phosphatidylcholine (PC) content in mature siliques along with an altered polyunsaturation level of the polar glycerolipids. In seeds, the acyl composition of triacylglycerol (TAG) was altered albeit not the content. These results indicate that plastidic LPAT1 plays an important role in reproductive growth and extraplastidic glycerolipid metabolism involving PC and TAG.
RESUMEN
The Kennedy pathway is a highly conserved de novo glycerolipid biosynthesis pathway in prokaryotes and eukaryotes. In Arabidopsis, LYSOPHOSPHATIDIC ACID ACYLTRANSFERASE 2 (LPAT2) was assumed to catalyze a crucial reaction step of the endoplasmic reticulum (ER)-localized Kennedy pathway because of lethality in the lpat2-1 knockout mutant. However, whether this lethal phenotype was due to the essential role of the Kennedy pathway or LPAT2 as the key enzyme of the Kennedy pathway was unclear. By creating non-lethal LPAT2-knockdown mutants in Arabidopsis, we found that LPAT2 is required for phospholipid content and plant development in vegetative and reproductive growth. Functional in vivo reporter assays revealed that LPAT2 was ubiquitously expressed and localized to the ER, where de novo phospholipid biosynthesis takes place. Intriguingly, our lipid analysis revealed that LPAT2 suppression had different effects among the organs examined: phospholipid levels were decreased both in leaves and flowers and the effect was more pronounced in flowers, a non-photosynthetic organ enriched with phospholipids. Although seed size was reduced in the LPAT2 suppression lines, no remarkable effect was observed in the lipid content of mature siliques. Our results show that LPAT2 is involved in the ER-localized Kennedy pathway, and suggest that its contribution to de novo phospholipid biosynthesis may have organ selectivity.