Methodology
An automated growth enclosure for metabolic labeling of Arabidopsis thaliana with 13C-carbon dioxide - an in vivo labeling system for proteomics and metabolomics research
1 Department of Horticultural Science, University of Minnesota, Saint Paul, USA
2 Department of Plant Biology, University of Minnesota, Saint Paul, USA
3 Microbial and Plant Genomics Institute, University of Minnesota, Saint Paul, USA
4 Saint Paul Apparatus Shop, University of Minnesota, Saint Paul, USA
5 Yeastern Biotech Co., Ltd. 6F, 23, Lane 169, Kang Ning St., Shijr, Taipei, Taiwan
6 Room S1-411, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, No. 1 West BeiChen road, ChaoYang district, Beijing, PR China
Proteome Science 2011, 9:9 doi:10.1186/1477-5956-9-9
Published: 10 February 2011Abstract
Background
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.
Results
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.
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 13C-labeling growth environment was built using readily available materials and software, it can be adapted easily to accommodate many different experimental designs.
