Leibniz-Institut für Pflanzengenetik und Kulturpflanzenforschung, 06466 Gatersleben, Germany.
Plant Physiol. 2013 Feb;161(2):583-93. doi: 10.1104/pp.112.210062. Epub 2012 Dec 11.
While often thought of as a smoking drug, tobacco (Nicotiana spp.) is now considered as a plant of choice for molecular farming and biofuel production. Here, we describe a noninvasive means of deriving both the distribution of lipid and the microtopology of the submillimeter tobacco seed, founded on nuclear magnetic resonance (NMR) technology. Our platform enables counting of seeds inside the intact tobacco capsule to measure seed sizes, to model the seed interior in three dimensions, to quantify the lipid content, and to visualize lipid gradients. Hundreds of seeds can be simultaneously imaged at an isotropic resolution of 25 µm, sufficient to assess each individual seed. The relative contributions of the embryo and the endosperm to both seed size and total lipid content could be assessed. The extension of the platform to a range of wild and cultivated Nicotiana species demonstrated certain evolutionary trends in both seed topology and pattern of lipid storage. The NMR analysis of transgenic tobacco plants with seed-specific ectopic expression of the plastidial phosphoenolpyruvate/phosphate translocator, displayed a trade off between seed size and oil concentration. The NMR-based assay of seed lipid content and topology has a number of potential applications, in particular providing a means to test and optimize transgenic strategies aimed at the manipulation of seed size, seed number, and lipid content in tobacco and other species with submillimeter seeds.
虽然烟草(Nicotiana spp.)通常被认为是一种吸烟用的毒品,但现在它被认为是用于分子农业和生物燃料生产的首选植物。在这里,我们描述了一种非侵入性的方法,基于核磁共振(NMR)技术,来获取脂质的分布和亚毫米级烟草种子的微观拓扑结构。我们的平台可以实现对完整烟草蒴果内种子的计数,以测量种子大小,对种子内部进行三维建模,量化脂质含量,并可视化脂质梯度。可以同时以 25 µm 的各向同性分辨率对数百个种子进行成像,足以评估每个种子。可以评估胚胎和胚乳对种子大小和总脂质含量的相对贡献。该平台扩展到一系列野生和栽培的 Nicotiana 物种,展示了种子拓扑结构和脂质储存模式的某些进化趋势。对具有种子特异性质体磷酸烯醇丙酮酸/磷酸转运蛋白异位表达的转基因烟草植物进行的 NMR 分析显示,种子大小和油浓度之间存在权衡。基于 NMR 的种子脂质含量和拓扑结构分析具有许多潜在的应用,特别是为测试和优化旨在操纵烟草和其他具有亚毫米种子的物种的种子大小、种子数量和脂质含量的转基因策略提供了一种手段。