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Quantitative and qualitative analysis of lipids in genetically modified potato tubers with varying rates of 14-3-3 protein synthesis

机译:14-3-3蛋白质合成速率不同的转基因马铃薯块茎中脂质的定量和定性分析

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In six transgenic lines of potatoes with the varying rates of 14-3-3 protein synthesis as well as in control cultivar Desiree the content and composition of the lipids extracted from the mature tubers from three years field trials (1998-2000) were analyzed. The transgenic lines J2 and J1 are both overexpressing gene encoding 14-3-3 protein. The J2 exhibited an overexpression of the protein 14-3-3 derived from pumpkin (Cucurbita pepo) cDNA and in J1 the 14-3-3 overexpression resulted from modifying of ADP-ribosylation factor synthesis. In the remaining lines, synthesis of the protein 14-3-3 was modified by the antisense technology. In tubers from 1998, the content of total lipids was within the range of 0.45-0.88% of tuber dry matter. The highest amount of fat was in tubers of line J2 (69% more than in the control). The content of lipids in tubers from subsequent years ranged from 0.36 to 0.63% of dry matter. Consistently the highest amount of fat was in tubers of line J2, however, the increase was very slight (8.6% more than in the control). The fractionation of lipids into polar and nonpolar fractions showed that all transgenic lines from field trials 1998 and 2000 contained more nonpolar lipids than the control (up to 270% in line J2). The percentage of nonpolar fractions in fats of tubers from all transgenes harvested in 1999 were similar, but they were higher than in tubers from the previous years, and they amounted to 44.4-49.1%. Chromatographic separation of methyl esters of fatty acids demonstrated that cis-alpha-linoleic acid was the main fatty acid present in potato tubers. This acid composed the biggest part of all lipids in G2 line. In the nonpolar fraction of lipids, palmitic acid followed by cis-alpha-linoleic acid showed the highest amounts.
机译:在六个具有14-3-3蛋白合成速率变化的转基因马铃薯中,以及在对照栽培品种Desiree中,对来自三年实地试验(1998-2000年)从成熟块茎中提取的脂质的含量和组成进行了分析。转基因品系J2和J1都过表达编码14-3-3蛋白的基因。 J2展示了南瓜(Cucurbita pepo)cDNA衍生的蛋白14-3-3的过表达,而J1中,修饰ADP-核糖基化因子合成导致14-3-3的过表达。在其余的品系中,通过反义技术修饰了蛋白质14-3-3的合成。 1998年以来的块茎中,总脂质的含量在块茎干物质的0.45-0.88%范围内。 J2系块茎中的脂肪含量最高(比对照高69%)。随后几年块茎中脂质的含量为干物质的0.36%至0.63%。一致的是,J2系块茎中的脂肪含量最高,但是增幅很小(比对照高8.6%)。脂质分成极性和非极性组分的分级显示,来自田间试验1998和2000的所有转基因品系比对照含有更多的非极性脂质(在J2系中高达270%)。 1999年收获的所有转基因块茎脂肪中非极性级分的百分率相似,但高于前几年的块茎脂肪,占44.4-49.1%。脂肪酸甲酯的色谱分离表明,顺式-α-亚油酸是马铃薯块茎中的主要脂肪酸。该酸构成了G2系中所有脂质的最大部分。在脂质的非极性组分中,棕榈酸紧随其后的是顺式-α-亚油酸,含量最高。

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