Abstract
In marine ecosystems, the avid binding of iron (Fe) to organic ligands influences Fe bioavailability in seaweed. This study aimed to elucidate Fe's biological availability to seaweed and develop a simple and rapid bioassay method as a new evaluation system. Undaria pinnatifida was used as a model seaweed species and the actual seaweed samples were collected using the 0.5 m × 0.5 m quadrat from the Mashike Bay area of Hokkaido, Japan. Chlorophyll fluorescence measurements were utilized as an index to evaluate the biological -effectiveness of Fe and compared with the results of culture tests based on growth. The effect of Fe content on media, pre-culture, concentrations and types of chelating and reducing agents in clearing solutions, cleaning time, Fe removal effect, and resistance to seaweed were systematically optimized to obtain the maximum efficacy of the washing solution. A bioassay was developed to evaluate the Fe environment by combining chlorophyll fluorescence measurements. The findings suggest that the tolerance of seaweeds to the wash solution is strongly influenced by the concentrations of the chelating and reducing agents than their types. Washing with 0.02 M Ti-Citrate/EDTA solution for 80 s was the most effective in terms of maximum Fe removal with minimum cell damage. The application of pre-culture and chemical pre-treatment methods under Fe deficiency to the culture strain confirmed the maximum reproducibility in the culture test. Finally, the developed method was applied to actual seaweed samples and was found to be applicable to many seaweed species. However, the method was less robust for some seaweed species and depended on the seaweed growth stage.
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