Physiological and cellular level responses of Enteromorpha spp. to chemical and thermal stress
The aims of this project were to investigate the cellular stress response (CSR) in Enteromorpha spp. and assess the potential of the Stress-70 protein (an indicator of the CSR) as a biomarker of pollutant exposure and acquired tolerance in Enteromorpha spp., compared with conventional physiological endpoints of toxicity. Cross-reactivity of a commercial Stress-70 antibody with E. intestinalis proteins was determined and used to develop an assay for Stress-70. Using this assay E. intestinalis was found to exhibit a typical heat shock response. Stress-70 proved to be a relatively insensitive biomarker of copper exposure and did not appear to be involved in copper tolerance, the genetic basis of which was investigated by growing E. intestinalis using a novel culturing technique. Although growth was variable, it provided a simple, consistent and sensitive measure of copper toxicity. The chlorophyll fluorescence parameter Fv/Fm was insensitive to copper exposure. Nutrient limitation enhanced copper toxicity and significantly impaired growth, Fv/Fm and Stress-70 production in E. intestinalis. In both copper ‘sensitive’ and ‘tolerant’ E. intestinalis, copper exposure did not affect the ability to raise a heat shock response. In ‘sensitive’ algae, copper and heat shock were additive stressors, with heat shock acting as a stronger inducer of Stress-70. Only heat shock affected 'tolerant' algae. Zinc was less toxic than copper but in contrast to copper studies, Stress-70 was a relatively sensitive indicator of zinc exposure, compared to Fv/Fm and growth. Studies of triazine herbicides revealed that on a molarity basis, Irgarol 1051 was more toxic to E. intestinalis than atrazine. Fv/Fm and growth were strongly affected by Irgarol exposure, but Stress-70 levels were unaltered by exposure to the herbicide. Fv/Fm and Stress-70 were poor in situ biomarkers of pollution, but another chlorophyll fluorescence parameter - complementary area - appeared to correlate with levels of organic pollution. Overall, Stress-70 was found not to be a useful biomarker of exposure to copper or triazines in E. intestinalis, or in situ pollution. However, the Stress-70 assay developed has a number of alternative applications and Enteromorpha spp. were deemed to be potentially useful in pollution monitoring with the selection of suitable biomarker responses.