Distinct functional microbial communities mediating the heterotrophic denitrification in response to the excessive Fe(II) stress in groundwater under wheat-rice stone and rock phosphate amendments.

Denitrifying microbial community can be utilized for eliminating nitrate and Fe(II) combined contamination in groundwater, while excessive amount of Fe(II) limit the process. Natural mineral can be additional substrate for the microbial growth, whereas how it influences the microbial community that mediating the denitrification coupling with Fe(II) oxidation and balancing inhibition of excessive Fe(II) on denitrification remain unclear. In the present study, we conducted a series of microcosm experiments to explore the denitrification and Fe(II) oxidation kinetic, and used RNA-based qPCR and DNA-based high-throughput sequencing to elucidate microbial diversity, co-occurrence and metabolic profiles amended by wheat-rice stone and rock phosphate. The results showed that both minerals could extensively improve and double the denitrification rates (2.0 ± 0.03 to 2.12 ± 0.13 times), decrease the nitrite accumulation and trigger the high resistance of the denitrifiers from the stress of Fe(II), whereas only wheat-rice stone with higher surface area increased the oxidation of Fe(II) (<10%). The addition of both minerals enhanced the microbial alpha-diversity, shaped the beta-diversity and co-occurrence network, and recovered the transcription of nitrate and nitrite reductase (Nar, Nap, NirS, NirK) from the Fe(II) inhibition. Accordingly, heterotroph Methyloversatilis sp., Methylotenra sp. might contribute to the denitrification under wheat-rice stone amendment, Denitratisoma sp. contribute to the denitrification for rock phosphate, and Fe oxidation was partially catalyzed by Dechloromonas sp. or abiotically by the nitrite/nitrous oxide. These findings would be helpful for better understanding the bioremediation of nitrate and Fe contaminated groundwater.

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Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.