Dissolved organic matter studies in enclosed systems: Application of Hydrophobic fractionation for the assessment of organic nitrogen dynamics

Abstract

The evolution of dissolved organic matter (DOM) in a non-axenic batch culture of the marine diatom Thalassiosira tumida was studied by hydrophobic fractionation during a three month experiment. DOM was fractionated with XAD-2 resin into hydrophobic (acid and neutral, “humic”) and hydrophilic fractions. The combined amino acid contents of unfractionated filtered seawater, XAD-fractions and particulate material were determined during the growth, stationary and degradation phases of the culture, and variations related to changes in dissolved organic nitrogen (DON) in XAD-fractions, dissolved inorganic nitrogen, algal and bacterial biomass. XAD-fractionation enabled the discrimination of simultaneously ocurring release and uptake of organic nitrogenous compounds: During the diatom growth there was a net increase of tolal DON concentrations, which was mostly accounted for by the hydrophilic fraction. A concurrent heterotrophic uptake of combined amino acids and other non-amino acid organic nitrogen was discernible by the decrease of their concentrations in the hydrophobic fractions. In the stationary phase, during the prevailing net consumption of total DON, the production of algal exudates could be detected in the hydrophobic fractions, while uptake mainly involved non-amino acid organic nitrogen from the hydrophilic fraction. During the degradation phase, after two months part of the particulate amino acid pool was transformed into hydrophilic DON, which in contrast to the stationary phase, was not adequate for supporting sustained bacterial growth. This suggests that the generation of recalcitrant substances may begin in the hydrophilic fraction of DOM. A slight increase of the hydrophobic acid fraction was indicative of the incipient formation of humic substances. XAD-2 was able to adsorb substances from fast changing DOM pools and thus should be a useful tool in studies concerned with phytoplankton and bacterial dynamics.