A high-throughput determination of metal concentrations in whole intact Arabidopsis thaliana seeds using synchrotron-based X-ray fluorescence spectroscopy.
The identification of genes involved in metal metabolism in plants requires the 'screening' of thousands of genetic variants. While inductively coupled plasma mass-spectroscopy has been used to identify variants with an altered total metal concentration, a more convenient high-throughput technique capable of examining individual seeds (or other tissues) would be useful. Here, the high brightness of synchrotron radiation has been utilised to examine relative metal concentrations in seeds of the genetically well characterised plant Arabidopsis thaliana. The relative concentrations of Mn, Fe, Ni, Cu and Zn in individual seeds were determined using a 500 microm x 500 microm beam. Metal concentrations were normally distributed, except where metal-containing dust contaminated the samples. Neither seed orientation nor genetic background (from three 'wild type' variants with different genetic lineages) had a significant affect on the Zn-normalised metal concentration. No advantages, such as the observation of tissue-specific metal accumulation, were obtained by using a 50 microm x 50 microm beam. A high-throughput proof-of-concept experiment was demonstrated that could be used to screen libraries of genetic variants for individuals with altered metal concentrations. Further work is required to standardise the technique before screening of libraries is possible.
Young
Lester W
LW
Agriculture and Agri-Food Canada, 107 Science Place, Saskatoon, SK, Canada S7N 0X2.
Westcott
Neil D
ND
Attenkofer
Klaus
K
Reaney
Martin J T
MJ
D000465:Algorithms; D017360:Arabidopsis / Q000737:chemistry*; D008670:Metals / Q000032:analysis*; D015203:Reproducibility of Results; D012639:Seeds / Q000737:chemistry*; D012680:Sensitivity and Specificity; D013048:Specimen Handling / Q000379:methods; D013052:Spectrometry, X-Ray Emission / Q000379:methods*; D017356:Synchrotrons
D008670:Metals