Department of Horticultural Science, University of Minnesota, Saint Paul, USA.
Proteome Sci. 2011 Feb 10;9(1):9. doi: 10.1186/1477-5956-9-9.
Labeling whole Arabidopsis (Arabidopsis thaliana) plants to high enrichment with 13C for proteomics and metabolomics applications would facilitate experimental approaches not possible by conventional methods. Such a system would use the plant's native capacity for carbon fixation to ubiquitously incorporate 13C from 13CO2 gas. Because of the high cost of 13CO2 it is critical that the design conserve the labeled gas.
A fully enclosed automated plant growth enclosure has been designed and assembled where the system simultaneously monitors humidity, temperature, pressure and 13CO2 concentration with continuous adjustment of humidity, pressure and 13CO2 levels controlled by a computer running LabView software. The enclosure is mounted on a movable cart for mobility among growth environments. Arabidopsis was grown in the enclosure for up to 8 weeks and obtained on average >95 atom% enrichment for small metabolites, such as amino acids and >91 atom% for large metabolites, including proteins and peptides.
The capability of this labeling system for isotope dilution experiments was demonstrated by evaluation of amino acid turnover using GC-MS as well as protein turnover using LC-MS/MS. Because this 'open source' Arabidopsis 13C-labeling growth environment was built using readily available materials and software, it can be adapted easily to accommodate many different experimental designs.
对拟南芥（Arabidopsis thaliana）植物进行 13C 高富集标记，用于蛋白质组学和代谢组学应用，将有助于采用常规方法无法进行的实验方法。这种系统将利用植物固碳的固有能力，将 13CO2 气体中的 13C 普遍掺入。由于 13CO2 的成本很高，因此设计必须节约标记气体。
设计并组装了一个完全封闭的自动化植物生长室，该系统可同时监测湿度、温度、压力和 13CO2 浓度，并通过运行 LabView 软件的计算机连续调整湿度、压力和 13CO2 水平。该室安装在可移动的小车上，可在不同的生长环境中移动。拟南芥在该室中生长了长达 8 周，平均获得了>95 原子%的小分子代谢物（如氨基酸）和>91 原子%的大分子代谢物（如蛋白质和肽）的富集。
通过使用 GC-MS 评估氨基酸周转率以及使用 LC-MS/MS 评估蛋白质周转率，证明了该标记系统进行同位素稀释实验的能力。由于这个“开源”的拟南芥 13C 标记生长环境是使用现成的材料和软件构建的，因此可以轻松适应许多不同的实验设计。
