Although denitrification-dependent chemolithotrophic sulfur oxidizers
proliferated in tsunami-deposited marine sediment with nitrate amendment,
their ecophysiological roles in biogeochemical carbon transfer are
not addressed. We employed time-resolved high-sensitivity 13C-bicarbonate probing of rRNA to unveil the carbon fixation and resulting
trophic relationship of the nitrate-amended sediment microorganisms.
Nitrate reduction and sulfur oxidation co-occurred along with significant
decreases in the 13CO2 and dissolved bicarbonate
concentrations for the first 4 days of the incubation, during which
the denitrification-dependent sulfur-oxidizing chemolithotrophs, i.e.,
the Sulfurimonas sp. HDS01 and Thioalkalispira sp. HDS22 relatives, and the sulfate-reducing heterotrophs, i.e.,
the Desulfobulbus spp. and Desulfofustis
glycolicus relatives, actively incorporated 13C. These indicated that the sulfur oxidizers and sulfate reducers
were tightly associated with each other through the direct carbon
transfer. Relatives of the fermentative Thalassomonas sediminis and the hydrolytic Pararheinheimera aquatica, in
addition to various sulfur-cycling microorganisms, significantly assimilated 13C at day 14. Although the incorporation of 13C
was not detected, a syntrophic volatile-fatty-acid oxidizer and hydrogenotrophic
methanogens significantly expressed their 16S rRNA molecules at day
21, indicating the metabolic activation of these final decomposers
under the latter nutrient-limited conditions. The results demonstrated
the nitrate-driven trophic association of sulfur-cycling microorganisms
and the subsequent microbial activation and diversification, triggering
the restoration of the marine ecosystem function.