Effect of metal stress on photosynthetic pigments in the Cu-hyperaccumulating lichens Cladonia humilis and Stereocaulon japonicum growing in Cu-polluted sites in Japan.

To understand the ecology and physiology of metal-accumulating lichens growing in Cu-polluted sites, we investigated lichens near temple and shrine buildings with Cu roofs in Japan and found that Stereocaulon japonicum Th. Fr. and Cladonia humilis (With.) J. R. Laundon grow in Cu-polluted sites. Metal concentrations in the lichen samples collected at some of these sites were determined by inductively coupled plasma mass spectroscopy (ICP-MS). UV-vis absorption spectra of pigments extracted from the lichen samples were measured, and the pigment concentrations were estimated from the spectral data using equations from the literature. Secondary metabolites extracted from the lichen samples were analyzed by high-performance liquid chromatography (HPLC) with a photodiode array detector. We found that S. japonicum and C. humilis are Cu-hyperaccumulating lichens. Differences in pigment concentrations and their absorption spectra were observed between the Cu-polluted and control samples of the 2 lichens. However, no correlation was found between Cu and pigment concentrations. We observed a positive correlation between Al and Fe concentrations and unexpectedly found high negative correlations between Al and pigment concentrations. This suggests that Al stress reduces pigment concentrations. The concentrations of secondary metabolites in C. humilis growing in the Cu-polluted sites agreed with those in C. humilis growing in the control sites. This indicates that the metabolite concentrations are independent of Cu stress.

Effect of copper stress on cup lichens Cladonia humilis and C. subconistea growing on copper-hyperaccumulating moss Scopelophila cataractae at copper-polluted sites in Japan.

We investigated lichen species in the habitats of the copper (Cu)-hyperaccumulating moss Scopelophila cataractae and found that the cup lichens Cladonia subconistea and C. humilis grow on this moss. We performed X-ray fluorescence and inductively coupled plasma mass (ICP-MS) analysis of lichen samples and measured the visible absorption spectra of the pigments extracted from the samples to assess the effect of Cu stress on the cup lichens. The chlorophyll a/b ratio and degradation of chlorophyll a to pheophytin a were calculated from the spectral data. X-ray fluorescence analysis indicated that Cu concentrations in cup lichens growing on S. cataractae were much higher than those in control samples growing on non-polluted soil. Moreover, Cu microanalysis showed that Cu concentrations in parts of podetia of C. subconistea growing on S. cataractae increased as the substrate (S. cataractae) was approached, whereas those of C. humilis growing on S. cataractae decreased as the substrate was approached. This reflects the difference in the route of Cu ions from the source to the podetia. Furthermore, ICP-MS analysis confirmed that C. subconistea growing on S. cataractae was heavily contaminated with Cu, indicating that this lichen is Cu tolerant. We found a significant difference between the visible absorption spectra of pigments extracted from the Cu-contaminated and control samples. Hence, the spectra could be used to determine whether a cup lichen is contaminated with Cu. Chlorophyll analysis showed that cup lichens growing on S. cataractae were affected by Cu stress. However, it also suggested that the areas of dead moss under cup lichens were a suitable substrate for the growth of the lichen. Moreover, it suggested that cup lichens had allolepathic effects on S. cataractae; it is likely that secondary metabolites produced by cup lichens inhibited moss growth.