Isotope fractionation of zinc in the paddy rice soil-water environment and the role of 2’deoxymugineic acid (DMA) as zincophore under Zn limiting conditions

Abstract

Non-traditional stable isotope systems are increasingly used to study micronutrient cycling and acquisition in terrestrial ecosystems. We previously proposed for zinc (Zn) a conceptual model linking observed isotope signatures and fractionations to biogeochemical processes occurring in the rice soil environment and we suggested that 2’deoxymugineic acid (DMA) could play an important role for rice during the acquisition of Zn when grown under Zn limiting conditions. This proposition was sustained by the extent and direction of isotope fractionation observed during the complexation of Zn with DMA synthesised in our laboratory. Here we report a new set of experimental data from field and laboratory studies designed to further elucidate the mechanisms controlling Zn isotope fractionation in the rice rhizosphere and the role of DMA. First, we present acidity (pKa) and complexation (logK) constants for DMA with H+ and Zn2+, respectively, using synthetic 2’deoxymugineic acid and show that they are significantly different from previously published data using isolates from plants. Our new set of thermodynamic data allows for a more accurate calculation of the formation of ZnDMA complexes over pH ranges typically found in the rhizosphere of flooded lowland rice soils and in rice plant compartments (xylem, phloem). We show that at pH > 6.5, Zn is fully complexed by DMA and at pH <4.5 fully dissociated. This has important implications, i.e. that in alkaline paddy soils, DMA can strip Zn from soil solids (organic and inorganic) and that ZnDMA complexes are stable at the root interface if the pH is alkaline and in the phloem and xylem of the rice shoot. Second, we present a new set of Zn isotope data in rice grown in alkaline soils with low Zn availability with and without Zn addition. We used two genotypes not tested to date, i.e. A69–1, tolerant to low Zn supply, and IR26, sensitive to low Zn supply. We confirm previous findings that, in contrast to observations with rice grown in hydroponic studies, no isotopically light Zn is taken up into the shoot irrespective of Zn fertilization and that isotopically heavier Zn is taken up by rice grown in soils with low Zn supply. Third, we determined the isotope fractionation isotherm for Zn during absorption on goethite, representing the iron phase (plaque) typically forming on rice roots, in acidic solution. The negative Zn isotope fractionation observed (i.e., Δ66Zngoethite-solution < 0, where Δ66Zngoethite-solution = (δ66Zn of goethite) – (δ66Zn of solution)) is in contrast to the positive isotope fractionation (Δ66Zngoethite-solution > 0) detected during adsorption of Zn on goethite in alkaline solutions or between root and soil solution in rice grown in alkaline soils. We show that removal of different Zn species from solution and changes in the Zn coordination control extent and direction of isotope fraction during adsorption. Using the new set of results and combining it with recent findings from the literature, we present a refined conceptual model linking biogeochemical processes to Zn isotope fractionation in the rice soil system. Our results confirm the importance of root induced chemical changes in the rhizosphere of rice growing in soils with low Zn availability, demonstrate the unique ability of isotope signatures to deconvolve geochemical processes and conditions in the plant-soil environment and support the hypothesis of an important role of DMA in Zn acquisition under Zn limiting conditions.