Influence of a low molecular weight metabolite (citrate) on the toxicity of cadmium and zinc to the unicellular green alga Selenastrum capricornutum: An exception to the free-ion model
Résumé
Toxicities of Cd and Zn to the chlorophyte Selenastrum capricornutum were determined in synthetic media buffered by 100 µmol*l -1 of NTA (FRAQ(NTA)) or citrate (FRAQ(CIT)), or in unbuffered FRAQUIL medium (FRAQ). Metal speciation in these exposure media was adjusted so as to maintain constant free concentrations of C 2+, Mg 2+ and Me(z+) even while the free Zn 2+ or Cd 2+ concentrations were varied. The growth response of the algae to variations in free Zn 2+ and free Cd 2+ was similar in FRAQ and FRAQ(NTA) media, but algal sensitivity to free Zn 2+ or free Cd 2+ increased markedly in FRAQ(crr) medium. Addition of Zn to FRAQ, FRAQ(NTA) or FRAQ(CIT) media (0.02-2 µmol*l -1 free Zn 2+) caused a decrease in the algal growth rate, but the algae remained in exponential growth just as long as in the control (72 h). Similar results were obtained for exposures to Cd in FRAQ or FRAQ(NTA) media (0.01-0.9 µmol*l -1 free Cd 2+), but additions of Cd to FRAQ(CIT) yielded anomalous growth curves (heightened sensitivity to Cd 2+ at low Cd levels; shortened period of exponential growth at high Cd levels). Short-term (6 min) intracellular uptake of radiolabelled 109Cd was measured in the three media containing the same free Cd 2+ concentration (0.25 µmol*l -1). Uptake of cadmium was more than 2-fold higher in FRAQ(CIT) than in the other two media. Under the same conditions, the algae could be shown to accumulate citrate; the measured uptake rate (83 pmol citrate m -2*s -1) was about 4 times greater than that of Cd in the presence of citrate (21 pmol Cd*m -2*s -1) - the citrate transporter would only have to be fooled once every four transport events to account for the enhanced cadmium uptake in the presence of citrate. Our results clearly show that the bioavailability of Cd and Zn diverges from the predictions of the Free-Ion Model in the presence of a low molecular weight metabolite such as citrate - accidental or piggy-back uptake of the metal-ligand complex across the biological membrane is a plausible explanation for this enhanced availability.Toxicities of Cd and Zn to the chlorophyte Selenastrum capricornutum were determined in synthetic media buffered by 100 µmol*l -1 of NTA (FRAQ NTA) or citrate (FRAQ CIT), or in unbuffered FRAQUIL medium (FRAQ). Metal speciation in these exposure media was adjusted so as to maintain constant free concentrations of Ca 2+, Mg 2+ and Me z+ even while the free Zn 2+ or Cd 2+ concentrations were varied. The growth response of the algae to variations in free Zn 2+ and free Cd 2+ was similar in FRAQ and FRAQ NTA media, but algal sensitivity to free Zn 2+ or free Cd 2+ increased markedly in FRAQ CIT medium. Addition of Zn to FRAQ, FRAQ NTA or FRAQ CIT media (0.02-2 µmol*l -1 free Zn 2+) caused a decrease in the algal growth rate, but the algae remained in exponential growth just as long as in the control (72 h). Similar results were obtained for exposures to Cd in FRAQ or FRAQ NTA media (0.01-0.9 µmol*l -1 free Cd 2+), but additions of Cd to FRAQ CIT yielded anomalous growth curves (heightened sensitivity to Cd 2+ at low Cd levels; shortened period of exponential growth at high Cd levels). Short-term (6 min) intracellular uptake of radiolabelled 109Cd was measured in the three media containing the same free Cd 2+ concentration (0.25 µmol*l -1). Uptake of cadmium was more than 2-fold higher in FRAQ CIT than in the other two media. Under the same conditions, the algae could be shown to accumulate citrate; the measured uptake rate (83 pmol citrate*m -2*s -1) was about 4 times greater than that of Cd in the presence of citrate (21 pmol Cd*m -2*s -1) - the citrate transporter would only have to be fooled once every four transport events to account for the enhanced cadmium uptake in the presence of citrate. Our results clearly show that the bioavailability of Cd and Zn diverges from the predictions of the Free-Ion Model in the presence of a low molecular weight metabolite such as citrate - accidental or piggy-back uptake of the metal-ligand complex across the biological membrane is a plausible explanation for this enhanced availability.