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Lipid and protein accumulation in developing seeds of three lupine species: Lupinus luteus L., Lupinus albus L., and Lupinus mutabilis Sweet.

Borek S, Pukacka S, Michalski K, Ratajczak L - J. Exp. Bot. (2009)

Bottom Line: Asparagine caused an increase in protein accumulation and simultaneously decreased the lipid content, but nitrate increased accumulation of both protein and lipid.The main fatty acid in yellow lupine cotyledons was linoleic acid, in white lupine it was oleic acid, and in Andean lupine it was both linoleic and oleic acids.The relationship between stimulation of lipid and protein accumulation by nitrate in developing lupine cotyledons and enhanced carbon flux through glycolysis caused by the inorganic nitrogen form is discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Physiology, Faculty of Biology, Adam Mickiewicz University, Umultowska 89, 61-614 Poznań, Poland. borek@amu.edu.pl

ABSTRACT
A comparative study was carried out on the dynamics of lipid accumulation in developing seeds of three lupine species. Lupine seeds differ in lipid content; yellow lupine (Lupinus luteus L.) seeds contain about 6%, white lupine (Lupinus albus L.) 7-14%, and Andean lupine (Lupinus mutabilis Sweet) about 20% of lipids by dry mass. Cotyledons from developing seeds were isolated and cultured in vitro for 96 h on Heller medium with 60 mM sucrose (+S) or without sucrose (-S). Each medium was additionally enriched with 35 mM asparagine or 35 mM NaNO3. Asparagine caused an increase in protein accumulation and simultaneously decreased the lipid content, but nitrate increased accumulation of both protein and lipid. Experiments with [1-14C]acetate and [2-14C]acetate showed that the decrease in lipid accumulation in developing lupine seeds resulted from exhaustion of lipid precursors rather than from degradation or modification of the enzymatic apparatus. The carbon atom from the C-1 position of acetate was liberated mainly as CO2, whereas the carbon atom from the C-2 position was preferentially used in anabolic pathways. The dominant phospholipid in the investigated lupine seed storage organs was phosphatidylcholine. The main fatty acid in yellow lupine cotyledons was linoleic acid, in white lupine it was oleic acid, and in Andean lupine it was both linoleic and oleic acids. The relationship between stimulation of lipid and protein accumulation by nitrate in developing lupine cotyledons and enhanced carbon flux through glycolysis caused by the inorganic nitrogen form is discussed.

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Electron micrographs of white lupine cotyledons isolated from seeds from developmental stage III, grown in vitro for 96 h. +S, medium with 60 mM sucrose; –S, medium without sucrose; +Asn, medium with 35 mM asparagine; +NO3–, medium with 35 mM NaNO3. CW, cell wall; M, mitochondrion; OB, oil body; S, starch; SP, storage protein; V, vacuole.
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fig6: Electron micrographs of white lupine cotyledons isolated from seeds from developmental stage III, grown in vitro for 96 h. +S, medium with 60 mM sucrose; –S, medium without sucrose; +Asn, medium with 35 mM asparagine; +NO3–, medium with 35 mM NaNO3. CW, cell wall; M, mitochondrion; OB, oil body; S, starch; SP, storage protein; V, vacuole.

Mentions: The total lipid level in in vitro grown cotyledons of each investigated species was higher in organs fed with sucrose (compare +S and –S; Fig. 4). Yellow and white lupine cotyledons grown in vitro on medium without sucrose (–S) contained less lipids than cotyledons used for preparation of in vitro culture (control in Fig. 4). In Andean lupine cotyledons the lipid level was higher (with and without sucrose) than in cotyledons used for preparation of in vitro culture (Fig. 4). Addition of asparagine to the medium (+S and −S) caused a decrease, whereas addition of NaNO3 caused an increase in total lipid level. The inhibitory effect of asparagine was the clearest and was statistically significant in yellow and Andean lupine cotyledons cultured on medium without sucrose (decrease of about 20.6% and 22.3%, respectively). In yellow, white, and Andean lupine cotyledons fed with sucrose and NaNO3, the lipid level was the highest among all the variants studied (increase of about 7.8, 8.6, and 10.2%, respectively; Fig. 4). The stimulatory effect of NaNO3 on lipid accumulation was clearly visible in the ultrastructure of cotyledon parenchyma cells (Figs 5–7). It was estimated that the highest number of oil bodies accumulating storage lipids in cotyledons of three lupine species was in organs cultured on medium with sucrose and with NaNO3. In cotyledons cultured on medium supplemented with asparagine, the number of lipid bodies was the lowest.


Lipid and protein accumulation in developing seeds of three lupine species: Lupinus luteus L., Lupinus albus L., and Lupinus mutabilis Sweet.

Borek S, Pukacka S, Michalski K, Ratajczak L - J. Exp. Bot. (2009)

Electron micrographs of white lupine cotyledons isolated from seeds from developmental stage III, grown in vitro for 96 h. +S, medium with 60 mM sucrose; –S, medium without sucrose; +Asn, medium with 35 mM asparagine; +NO3–, medium with 35 mM NaNO3. CW, cell wall; M, mitochondrion; OB, oil body; S, starch; SP, storage protein; V, vacuole.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC2724698&req=5

fig6: Electron micrographs of white lupine cotyledons isolated from seeds from developmental stage III, grown in vitro for 96 h. +S, medium with 60 mM sucrose; –S, medium without sucrose; +Asn, medium with 35 mM asparagine; +NO3–, medium with 35 mM NaNO3. CW, cell wall; M, mitochondrion; OB, oil body; S, starch; SP, storage protein; V, vacuole.
Mentions: The total lipid level in in vitro grown cotyledons of each investigated species was higher in organs fed with sucrose (compare +S and –S; Fig. 4). Yellow and white lupine cotyledons grown in vitro on medium without sucrose (–S) contained less lipids than cotyledons used for preparation of in vitro culture (control in Fig. 4). In Andean lupine cotyledons the lipid level was higher (with and without sucrose) than in cotyledons used for preparation of in vitro culture (Fig. 4). Addition of asparagine to the medium (+S and −S) caused a decrease, whereas addition of NaNO3 caused an increase in total lipid level. The inhibitory effect of asparagine was the clearest and was statistically significant in yellow and Andean lupine cotyledons cultured on medium without sucrose (decrease of about 20.6% and 22.3%, respectively). In yellow, white, and Andean lupine cotyledons fed with sucrose and NaNO3, the lipid level was the highest among all the variants studied (increase of about 7.8, 8.6, and 10.2%, respectively; Fig. 4). The stimulatory effect of NaNO3 on lipid accumulation was clearly visible in the ultrastructure of cotyledon parenchyma cells (Figs 5–7). It was estimated that the highest number of oil bodies accumulating storage lipids in cotyledons of three lupine species was in organs cultured on medium with sucrose and with NaNO3. In cotyledons cultured on medium supplemented with asparagine, the number of lipid bodies was the lowest.

Bottom Line: Asparagine caused an increase in protein accumulation and simultaneously decreased the lipid content, but nitrate increased accumulation of both protein and lipid.The main fatty acid in yellow lupine cotyledons was linoleic acid, in white lupine it was oleic acid, and in Andean lupine it was both linoleic and oleic acids.The relationship between stimulation of lipid and protein accumulation by nitrate in developing lupine cotyledons and enhanced carbon flux through glycolysis caused by the inorganic nitrogen form is discussed.

View Article: PubMed Central - PubMed

Affiliation: Department of Plant Physiology, Faculty of Biology, Adam Mickiewicz University, Umultowska 89, 61-614 Poznań, Poland. borek@amu.edu.pl

ABSTRACT
A comparative study was carried out on the dynamics of lipid accumulation in developing seeds of three lupine species. Lupine seeds differ in lipid content; yellow lupine (Lupinus luteus L.) seeds contain about 6%, white lupine (Lupinus albus L.) 7-14%, and Andean lupine (Lupinus mutabilis Sweet) about 20% of lipids by dry mass. Cotyledons from developing seeds were isolated and cultured in vitro for 96 h on Heller medium with 60 mM sucrose (+S) or without sucrose (-S). Each medium was additionally enriched with 35 mM asparagine or 35 mM NaNO3. Asparagine caused an increase in protein accumulation and simultaneously decreased the lipid content, but nitrate increased accumulation of both protein and lipid. Experiments with [1-14C]acetate and [2-14C]acetate showed that the decrease in lipid accumulation in developing lupine seeds resulted from exhaustion of lipid precursors rather than from degradation or modification of the enzymatic apparatus. The carbon atom from the C-1 position of acetate was liberated mainly as CO2, whereas the carbon atom from the C-2 position was preferentially used in anabolic pathways. The dominant phospholipid in the investigated lupine seed storage organs was phosphatidylcholine. The main fatty acid in yellow lupine cotyledons was linoleic acid, in white lupine it was oleic acid, and in Andean lupine it was both linoleic and oleic acids. The relationship between stimulation of lipid and protein accumulation by nitrate in developing lupine cotyledons and enhanced carbon flux through glycolysis caused by the inorganic nitrogen form is discussed.

Show MeSH
Related in: MedlinePlus