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Sulfate nutrient-deprivation in Chlamydomonas reinhardtii brings about prompt degradation of Rubisco and a concomitant substantial accumulation of starch. These changes precede hydrogen ( H 2 ) photoevolution by the cells. The cause-and-effect relationship between Rubisco loss, starch accumulation and subsequent H 2 -photoevolution in C. reinhardtii, and the role of illumination for these changes to occur, was investigated in this work. A Rubisco-less and acetate-requiring mutant of C. reinhardtii (CC2653) was employed as a tool in this investigation and compared to the wild type (WT) in terms of protein and starch metabolic flux and H 2 -evolution upon sulfur deprivation. Results showed a prompt Rubisco degradation and concomitant 10-fold starch accumulation in the WT in the light, which was completed within 48 h of S-deprivation. This was followed by a regulated starch degradation and concomitant H 2 -photoevolution, which lasted for up to 120 h in S-deprivation. This massive flux of primary metabolites (protein and starch) did not occur in the dark in the WT, suggesting a strictly light-dependent and integrated process in metabolite rearrangement and H 2 -photoevolution in C. reinhardtii. The Rubisco-less CC2653 mutant failed to accumulate starch upon S-deprivation in the light or dark and also failed to evolve H 2 gas. These results suggested a temporal cause-and-effect relationship between the light-dependent catabolism of Rubisco and starch accumulation, and the subsequent ability of the cell to perform a light-dependent starch degradation and H 2 -photoevolution. The regulated starch breakdown in the light apparently provides the endogenous substrate that supports H 2 -evolution, both by feeding electrons into the plastoquinone pool in chloroplasts, and indirectly by sustaining mitochondrial respiration for the maintenance of anaerobiosis in the cell.

Photosynthetic and antioxidant responses following exposure to either ultraviolet-A or ultraviolet-B were contrasted in two species of the unicellular green alga, DUNALIELLA: Species selection was based on the ability of Dunaliella bardawil (UTEX 2538) to accumulate inter-thylakoid beta-carotene when subjected to environmental stress while Dunaliella salina (UTEX 200) lacks this ability. Cells were cultured in high and low levels of visible light (150 and 35 micro mol photons m(-2 )s(-1), respectively) and then either ultraviolet-A (320-400 nm) or ultraviolet-B (290-320 nm) was added to visible light for 24-h exposure. A potassium chromate solution was found to be an ideal screen for removal of ultraviolet-A and ultraviolet-C from ultraviolet-B radiation. There were no significant changes in photosynthetic or antioxidant parameters following exposure to ultraviolet-B. Ultraviolet-A exposure significantly decreased photosynthetic parameters (>70% decrease in Fv/Fm and the ratio of light-limited to light-saturated photosynthesis in low beta-carotene cells) and resulted in 50% increases in ascorbate peroxidase activity and ascorbate concentrations. The results suggest exposure to ultraviolet-A (but not ultraviolet-B) directly affects photosynthesis, observed as a loss of photosystem II electron transport efficiency and increased radical formation. This research indicates that the accumulated beta-carotene in D. bardawil prevents UV-related photosynthetic damage through blue-light/ultraviolet-A absorption (supported by trends observed for antioxidant enzyme responses).

The impact of herbicide exposure on nontarget vegetation within agroecosystems has sparked extensive research that revealed that current pesticide registration guidelines may be inadequate at predicting the effects of herbicides on wild plants and habitats. This study extends the current interest by presenting three experiments highlighting some of the limitations to current phytotoxicity testing guidelines. Several crops and wild plant species were grown under greenhouse conditions following standard protocol for phytotoxicity testing. Plants were sprayed with five different herbicides at the four- to six-leaf stage, and biomass was recorded at 28 d after spray. Results showed that current regulatory protocol will likely underestimate herbicide phytotoxicity if testing does not include data for the complete tank-mix formulation. The present study also showed that the range in herbicide sensitivity among cultivars of the same crop can be quite extensive and that, depending on the cultivar included in a risk assessment, conclusions regarding the phytotoxicity of any given herbicide may differ. Although no significant differences in sensitivity were found between crops and related wild species, results revealed that current guidelines are too rigid in terms of species selection. Considering the variability among crop cultivars, coupled with the ecological importance and the ease of germination of many noncrop plant species, pesticide regulatory guidelines would be improved if wild species were included in testing. Findings of the present study indicate that current pesticide regulatory guidelines require modifications to ensure a more accurate assessment of herbicide effects on nontarget plant species.