VERFAHREN ZUR ERHÖHUNG DES GESAMTÖLGELHALTES IN ÖLPFLANZEN
Vigeolas, Hélène ; ; et al
DE) Die Erfindung betrifft Verfahren zur Erhöhung des Gesamtölgehaltes und/oder des Gehalts an Glycerol-3-Phosphat in transgenen Ölpflanzen, die mindestens 20 Gew-% Ölsäure bezogen auf den ... [more ▼]
DE) Die Erfindung betrifft Verfahren zur Erhöhung des Gesamtölgehaltes und/oder des Gehalts an Glycerol-3-Phosphat in transgenen Ölpflanzen, die mindestens 20 Gew-% Ölsäure bezogen auf den Gesamtfettsäuregehalt enthalten, bevorzugt in pflanzlichen Samen, durch Expression von Glycerol-3-phosphatdehydrogenasen (G3PDH) aus Hefen, bevorzugt aus Saccharomyces cerevisiae. Vorteilhaft wird das im Verfahren gewonnene Öl und/oder die freien Fettsäure Polymeren, Nahrungsmitteln, Futtermitteln, Kosmetika, Pharmazeutika oder Produkten mit industriellen Anwendungen zugesetzt. [less ▲]Detailed reference viewed: 16 (3 ULg)
Increasing seed oil content in oil-seed rape (Brassica napus L.) by over-expression of a yeast glycerol-3-phosphate dehydrogenase under the control of a seed-specific promoter.
Vigeolas, Hélène ; ; et al
in Plant Biotechnology Journal (2007), 5(3), 431-41
Previous attempts to manipulate oil synthesis in plants have mainly concentrated on the genes involved in the biosynthesis and use of fatty acids, neglecting the possible role of glycerol-3-phosphate ... [more ▼]
Previous attempts to manipulate oil synthesis in plants have mainly concentrated on the genes involved in the biosynthesis and use of fatty acids, neglecting the possible role of glycerol-3-phosphate supply on the rate of triacylglycerol synthesis. In this study, a yeast gene coding for cytosolic glycerol-3-phosphate dehydrogenase (gpd1) was expressed in transgenic oil-seed rape under the control of the seed-specific napin promoter. It was found that a twofold increase in glycerol-3-phosphate dehydrogenase activity led to a three- to fourfold increase in the level of glycerol-3-phosphate in developing seeds, resulting in a 40% increase in the final lipid content of the seed, with the protein content remaining substantially unchanged. This was accompanied by a decrease in the glycolytic intermediate dihydroxyacetone phosphate, the direct precursor of glycerol-3-phosphate dehydrogenase. The levels of sucrose and various metabolites in the pathway from sucrose to fatty acids remained unaltered. The results show that glycerol-3-phosphate supply co-limits oil accumulation in developing seeds. This has important implications for strategies that aim to increase the overall level of oil in commercial oil-seed crops for use as a renewable alternative to petrol. [less ▲]Detailed reference viewed: 46 (4 ULg)
Lipid storage metabolism is limited by the prevailing low oxygen concentrations within developing seeds of oilseed rape.
Vigeolas, Hélène ; ; et al
in Plant Physiology (2003), 133(4), 2048-60
The aim of this study was to investigate whether endogenous restrictions in oxygen supply are limiting for storage metabolism in developing oilseed rape (Brassica napus) seeds. Siliques were studied 30 d ... [more ▼]
The aim of this study was to investigate whether endogenous restrictions in oxygen supply are limiting for storage metabolism in developing oilseed rape (Brassica napus) seeds. Siliques were studied 30 d after flowering, when rapid lipid accumulation is occurring in the seeds. (a). By using microsensors, oxygen concentrations were measured within seeds and in the silique space between seeds. At ambient external oxygen (21% [v/v]) in the light, oxygen fell to 17% (v/v) between and 0.8% (v/v) within seeds. A step-wise reduction of the external oxygen concentration led within 2 h to a further decrease of internal oxygen concentrations, and a step-wise increase of the external oxygen concentration up to 60% (v/v) resulted in an increase in internal oxygen that rose to 30% (v/v) between and 8% (v/v) within seeds. (b). The increase in oxygen levels in the seeds was accompanied by a progressive increase in the levels of ATP, UTP, and the ATP to ADP and UTP to UDP ratios over the entire range from 0% to 60% (v/v) external oxygen. (c). To investigate metabolic fluxes in planta, 14C-sucrose was injected into seeds, which remained otherwise intact within their siliques. The increase in oxygen in the seeds was accompanied by a progressive increase in the rate of lipid (including triacylglycerol), protein and cell wall synthesis, and an increase in glycolytic flux over a range from sub- to superambient oxygen concentrations. In contrast to lipid synthesis, starch synthesis was not significantly increased at superambient oxygen levels. The levels of fermentation products such as lactate and glycerol-3P increased only at very low (0%-4% [v/v]) external oxygen concentrations. (d). When 14C-acetate or 14C-acetyl-coenzyme A (CoA) was injected into seeds, label incorporation into triacylglycerol progressively increased over the whole range of external oxygen concentrations from 0% to 60% (v/v). (e). Stimulation of lipid synthesis was accompanied by an increase in sugar levels and a decrease in the levels of hexose-phosphates and acetyl-CoA, indicating sucrose unloading and the use of acetyl-CoA as possible regulatory sites. (f). Increased lipid synthesis was also accompanied by an increase in the maximal activities of invertase and diacylglycerol acyltransferase. (g). The developmental shift from starch to lipid storage between 15 and 45 d after flowering was accompanied by an increase in the seed energy state. (h). The results show that at ambient oxygen levels, the oxygen supply is strongly limiting for energy metabolism and biosynthetic fluxes in growing rape seeds, affecting lipid synthesis more strongly than starch synthesis. The underlying mechanisms and implications for strategies to increase yield and storage product composition in oilseed crops are discussed. [less ▲]Detailed reference viewed: 17 (0 ULg)