An automated growth enclosure for metabolic labeling of Arabidopsis thaliana with C-13-carbon dioxide - an in vivo labeling system for proteomics and metabolomics research

作者：Chen, Wen-Ping; Yang, Xiao-Yuan; Harms, Geoffrey L.; Gray, William M.; Hegeman, Adrian D.^*; Cohen, Jerry D.

来源：Proteome Science, 2011, 9: 9.

DOI：10.1186/1477-5956-9-9

• CHROMATOGRAPHY-MASS SPECTROMETRY
• QUANTITATIVE-ANALYSIS
• AMINO-ACIDS
• ISOTOPE-DILUTION
• STABLE-ISOTOPES
• FLUX ANALYSIS
• (CO2)-C-13
• TURNOVER
• LEAVES
• PLANTS

Background: Labeling whole Arabidopsis (Arabidopsis thaliana) plants to high enrichment with C-13 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 C-13 from (CO2)-C-13 gas. Because of the high cost of (CO2)-C-13 it is critical that the design conserve the labeled gas.
Results: A fully enclosed automated plant growth enclosure has been designed and assembled where the system simultaneously monitors humidity, temperature, pressure and (CO2)-C-13 concentration with continuous adjustment of humidity, pressure and (CO2)-C-13 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.
Conclusion: 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 C-13-labeling growth environment was built using readily available materials and software, it can be adapted easily to accommodate many different experimental designs.

• 出版日期2011-2-10
• 单位中国科学院遗传与发育生物学研究所