Stable isotopically labelled organisms have found wide application in life science research including plant physiology, plant stress and defense as well as metabolism related sciences. Therefore, the reproducible production of plant material enriched with stable isotopes such as 13C and 15N is of considerable interest. A high degree of enrichment (> 96 atom %) with a uniformly distributed isotope (global labelling) is accomplished by a continuous substrate supply during plant growth/cultivation. In the case of plants, 13C-labelling can be achieved by growth in 13CO2(g) atmosphere while global 15N-labelling needs 15N- containing salts in the watering/nutrient solution. Here, we present a method for the preparation of 13C and 15N-labelled plants by the use of closed growth chambers and hydroponic nutrient supply. The method is exemplified with durum wheat. In total, 330 g of globally 13C- and 295 g of 15N-labelled Triticum durum wheat was produced during 87 cultivation days. For this, a total of 3.88 mol of 13CO2(g) and 58 mmol of 15N were consumed. The degree of enrichment was determined by LC-HRMS and ranged between 96 and 98 atom % for 13C and 95–99 atom % for 15N, respectively. Additionally, the isotopically labelled plant extracts were successfully used for metabolome-wide internal standardisation of native T.durum plants. Application of an isotope-assisted LC-HRMS workflow enabled the detection of 652 truly wheat-derived metabolites out of which 143 contain N. A reproducible cultivation which makes use of climate chambers and hydroponics was successfully adapted to produce highly enriched, uniformly 13C- and 15N-labelled wheat. The obtained plant material is suitable to be used in all kinds of isotope-assisted research. The described technical equipment and protocol can easily be applied to other plants to produce 13C-enriched biological samples when the necessary specific adaptations e.g. temperature and light regime, as well as nutrient supply are considered. Additionally, the 15N-labelling method can also be carried out under regular glasshouse conditions without the need for customised atmosphere.

中文翻译：

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使用定制的生长室制备统一标记的 13C 和 15N 植物

稳定同位素标记的生物体已广泛应用于生命科学研究，包括植物生理学、植物胁迫和防御以及代谢相关科学。因此，富含稳定同位素（如 13C 和 15N）的植物材料的可重复生产具有相当大的意义。通过在植物生长/培养过程中连续提供底物，实现同位素分布均匀（全局标记）的高度富集（> 96 atom %）。在植物的情况下，13C 标记可以通过在 13CO2(g) 大气中生长来实现，而全球 15N 标记需要在浇水/营养液中含有 15N 的盐。在这里，我们提出了一种通过使用封闭的生长室和水培营养供应来制备 13C 和 15N 标记植物的方法。该方法以硬质小麦为例。在 87 个种植日期间，总共生产了 330 克全球 13C 和 295 克 15N 标记的硬粒小麦。为此，总共消耗了 3.88 mol 的 13CO2(g) 和 58 mmol 的 15N。富集程度由 LC-HRMS 测定，13C 和 15N 分别在 96 和 98 原子％之间和 95-99 原子％之间。此外，同位素标记的植物提取物成功地用于天然硬粒小麦植物的代谢组范围内标准化。同位素辅助 LC-HRMS 工作流程的应用能够检测 652 种真正源自小麦的代谢物，其中 143 种含有 N。利用气候室和水培的可重复栽培成功地适应了生产高度富集、均匀的 13C 和15N 标记的小麦。所得植物材料适用于各种同位素辅助研究。当考虑必要的特定适应（例如温度和光照条件以及营养供应）时，所描述的技术设备和协议可以很容易地应用于其他植物以生产富含 13C 的生物样品。此外，15N 标记方法也可以在常规温室条件下进行，无需定制气氛。