Molecular response of Anabaena flos-aquae to differing concentrations of phosphorus:: A combined Fourier transform infrared and X-ray microanalytical study

The response of Anabaena flos-aquae to varying levels of P availability was determined using the combined techniques of Fourier transform infrared (FTIR) microspectroscopy and energy-dispersive X-ray microanalysis (EDXRMA). Batch cultures of Anabaena were grown at initial P concentrations of 50 mu g (low-P culture), 500 mu g (intermediate-P) and 5000 mu g (high-P) (PO(4)-P L(-1)) and were monitored for total biovolume, chlorophyll a, media nutrients (PO(4)-P and NO(3)-N), cellular P quota (EDXRMA) and carbon allocation (FTIR spectroscopy). The high-P culture showed a sixfold increase in total biovolume over the sampling period. Phosphorus in the media remained at more than similar to 4000 mu g L(-1) and intracellular P concentration (P quota) as determined by EDXRMA showed no decline, remaining at more than 0.20% dry weight (DW). The intermediate-P culture showed a similar increase in total biovolume, but P concentrations in the media fell to similar to 20 mu g L(-1), and this was reflected in a decline in the cellular P quota from 0.24% to 0.06% DW. Although the high- and intermediate-P treatments differed markedly in both internal and external P, analysis of the FTIR spectra from the two treatments, using both hierarchical cluster analysis (HCA) and principal component analysis (PCA), indicated no difference in carbon allocation. Cells from the low-P treatment showed strong evidence for P deficiency. P concentrations in the media were undetectable (< 5 mu g L(-1)), total biovolume was much reduced, and P quotas were low, falling from 0.08% to 0.01% DW. HCA and PCA clearly separated FTIR spectra from low-P cells from those of intermediate- and high-P cultures, with low-P cells having increased absorbance in the region of the spectra associated with carbohydrate. Both FTIR spectroscopy and EDXRMA have the resolution to study the elemental and macromolecular composition of single species within mixed phytoplankton populations and the combined use of these techniques has considerable potential for the study of cyanobacterial responses to environmental stress.