Department of Biological Sciences, Center for Plant Lipid Research, University of North Texas, Denton, TX 76203, USA.
Biochimie. 2014 Jan;96:28-36. doi: 10.1016/j.biochi.2013.08.010. Epub 2013 Aug 22.
Targeted increases in monounsaturated (oleic acid) fatty acid content of refined cottonseed oil could support improved human nutrition and cardiovascular health. Genetic modifications of cottonseed fatty acid composition have been accomplished using several different molecular strategies. Modification of oleic acid content in cottonseed embryos using a dominant-negative protein approach, while successful in effecting change in the desired fatty acid composition, resulted in reduced oil content and seed viability. Here these changes in fatty acid composition were associated with changes in dominant molecular species of triacylglycerols (TAGs) and their spatial distributions within embryo tissues. A combination of mass spectrometry (MS)-based lipidomics approaches, including MS imaging of seed cryo-sections, revealed that cotton embryos expressing a non-functional allele of a Brassica napus delta-12 desaturase showed altered accumulation of TAG species, especially within cotyledonary tissues. While lipid analysis of seed extracts could demonstrate detailed quantitative changes in TAG species in transgenics, the spatial contribution of metabolite compartmentation could only be visualized by MS imaging. Our results suggest tissue-specific differences in TAG biosynthetic pathways within cotton embryos, and indicate the importance of considering the location of metabolites in tissues in addition to their identification and quantification when developing a detailed view of cellular metabolism.
提高精炼棉籽油中单不饱和(油酸)脂肪酸含量可能有助于改善人类营养和心血管健康。已经使用几种不同的分子策略对棉籽油脂肪酸组成进行了遗传修饰。使用显性负蛋白方法修饰棉籽油胚中的油酸含量,虽然在改变所需脂肪酸组成方面取得了成功,但导致油含量和种子活力降低。在这里,脂肪酸组成的这些变化与三酰基甘油 (TAG) 的主要分子种类的变化及其在胚组织内的空间分布有关。基于质谱 (MS) 的脂质组学方法的组合,包括对种子冷冻切片的 MS 成像,揭示了表达 Brassica napus delta-12 去饱和酶无功能等位基因的棉花胚胎显示出 TAG 种类积累的改变,特别是在子叶组织内。虽然种子提取物的脂质分析可以证明转基因中 TAG 种类的详细定量变化,但只有通过 MS 成像才能观察到代谢物区室化的空间贡献。我们的结果表明,棉籽油胚中的 TAG 生物合成途径存在组织特异性差异,并表明在开发细胞代谢的详细视图时,除了鉴定和定量外,还需要考虑代谢物在组织中的位置。