WAX INDUCER1/SHINE1 (WIN1) belongs to the AP2/EREBP transcription factor family and plays an important role in wax and cutin accumulation in plants. Here we show that BnWIN1 from Brassica napus (Bn) has dual functions in wax accumulation and oil synthesis. Overexpression (OE) of BnWIN1 led to enhanced wax accumulation and promoted growth without adverse effects on oil synthesis under salt stress conditions. Lipid profiling revealed that BnWIN1-OE plants accumulated more waxes with elevated C29-alkanes, C31-alkanes, C28-alcohol, and C29-alcohol relative to wild type (WT) under salt stress. Moreover, overexpression of BnWIN1 also increased seed oil content under normal growth conditions. BnWIN1 directly bound to the promoter region of genes encoding biotin carboxyl carrier protein 1 (BCCP1), glycerol-3-phosphate acyltransferase 9 (GPAT9), lysophosphatidic acid acyltransferase 5 (LPAT5), and diacylglycerol acyltransferase 2 (DGAT2) involved in the lipid anabolic process. Overexpression of BnWIN1 resulted in upregulated expression of numerous genes involved in de novo fatty acid synthesis, wax accumulation, and oil production. The results suggest that BnWIN1 is a transcriptional activator to regulate the biosynthesis of both extracellular and intracellular lipids.

Plastid-localized glycerol-3-phosphate acyltransferase (ATS1) catalyzes the first-step reaction in glycerolipid assembly through transferring an acyl moiety to glycerol-3-phosphate (G3P) to generate lysophosphatidic acid (LPA), an intermediate in lipid metabolism. The effect of ATS1 overexpression on glycerolipid metabolism and growth remained to be elucidated in plants, particularly oil crop plants. Here, we found that overexpression of BnATS1 from Brassica napus enhanced plant growth and prokaryotic glycerolipid biosynthesis. BnATS1 is localized in chloroplasts and an in vitro assay showed that BnATS1 had acylation activity toward glycerol 3-phosphate to produce LPA. Lipid profiling showed that overexpression of BnATS1 led to increases in multiple glycerolipids including phosphatidylglycerol (PG), monogalactosyldiacylglycerol (MGDG), phosphatidylcholine (PC), and phosphatidylinositol (PI), with increased polyunsaturated fatty acids. Moreover, increased MGDG was attributed to the elevation of 34:6- and 34:5-MGDG, which were derived from the prokaryotic pathway. These results suggest that BnATS1 promotes accumulation of polyunsaturated fatty acids in cellular membranes, thus enhances plant growth under low-temperature conditions in Brassica napus.

Phosphatidic acid (PA) is an important signal molecule in various biological processes including osmotic stress. Lysophosphatidic acid acyltransferase (LPAT) acylates the sn-2 position of the glycerol backbone of lysophosphatidic acid (LPA) to produce PA. The role of LPAT2 and its PA in osmotic stress response remains elusive in plants. Here we showed that LPAT2-derived PA is important for salt and drought stress tolerance in rice. Rice LPAT2 was localized to the endoplasmic reticulum (ER) to catalyze the PA synthesis. The LPAT2 transcript was induced by osmotic stress such as high salinity and water deficit. To reveal its role in osmotic stress response, an LPAT2 knockdown mutant, designated lpat2, was isolated from rice, which contained a reduced PA level relative to wild type (WT) plants under salt stress and water deficit. The lpat2 mutant was more susceptible to osmotic stress and less sensitive to abscisic acid (ABA) than that of WT, which was recovered by either PA supplementation or genetic LPAT2 complementation. Moreover, suppressed LPAT2 also led to a large number of differentially expressed genes (DEGs) involved in diverse processes, particularly, in ABA response, kinase signaling, and ion homeostasis in response to salt stress. Together, LPAT2-produced PA plays a positive role in osmotic tolerance through mediating ABA response, which leads to transcriptional alteration of genes related to ABA response, protein kinase signaling, and ion homeostasis.