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This work examines nutritional influence on fungal colony growth and biomass distribution in response to toxic metals. In low-substrate solid medium, 0.1 mM Cd, Cu and Zn caused a decrease in radial expansion of both Trichoderma viride and Rhizopus arrhizus. However, as the amount of available carbon source (glucose) increased, the apparent toxicity of the metals decreased. These metals also affected the overall length of the fungal mycelium and branching patterns. In low-nutrient conditions, T. viride showed a decrease in overall mycelial length and number of branches in response to Cu, resulting in an extremely sparsely branched colony. Conversely, although Cd also reduced overall mycelial length to about one-third of the control length, the number of branches decreased only slightly which resulted in a highly branched colony with many aberrant features. Cu and Cd induced similar morphological changes in R. arrhizus. A large-scale mycelial-mapping technique showed that disruption of normal growth by Cu and Cd resulted in altered biomass distribution within the colony. When grown on metal-free low-substrate medium, T. viride showed an even distribution of biomass within the colony with some allocation to the periphery. However, Cu caused most of the biomass to be allocated to the colony periphery, while in the presence of Cd, most biomass was located at the interior of the colony. These results imply that such alterations of growth and resource allocation by Cu and Cd may influence success in locating nutrients as well as survival, and that these metals have individual and specific effects on the growing fungus.

To directly define vacuolar role(s) in metal detoxification, we have examined the responses of vacuole-deficient mutants of Saccharomyces cerevisiae to several potentially toxic metals known to be mainly detoxified in the cytosol (Cu, Cd) or the vacuole (Co, Mn, Ni, Zn). Three mutants, deficient in targeting of vacuolar proteins, were used with JSR18 delta 1 being devoid of any vacuole-like structure while ScVatB and ScVatC were deficient in specific protein subunits of the V-ATPase. The results obtained show that the absence of a vacuole or a functional vacuolar H(+)-ATPase was associated with increased sensitivity and a largely decreased capacity of the vacuole-deficient strains to accumulate Zn, Mn, Co and Ni, confirming an essential role for the vacuole in detoxification of these metals. In addition, the lack of vacuolar involvement in detoxification of Cu and Cd was confirmed since these metals did not exhibit increased toxicity towards the vacuolar mutants nor were there significant differences in Cu or Cd accumulation between parental and mutant strains.

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