The nitrogen cycle is an indispensable part of the biogeochemical cycle, and the reactions that occur in the ocean oxygen minimum zone (OMZ) mediate much of the loss of nitrogen from oceans worldwide. Here, nitrate-utilizing bacteria were isolated from the water column at 17 stations within the OMZ of the Changjiang (Yangtze River) Estuary using selective media and a culture-dependent method. The microbial diversity, nitrogen metabolism and nitrate reduction test of culturable heterotrophic bacteria were examined. A total of 164 isolates were obtained; they were mostly affiliated with Proteobacteria (81.1%), Actinobacteria (5.5%), Bacteroidetes (12.3%), and Firmicutes (0.6%). Pseudomonas aeruginosa, Sphingobium naphthae, and Zunongwangia profunda were found at most stations. Among 24 tested representative strains, 8 were positive for nitrate reduction; they belonged to genera Aurantimonas, Halomonas, Marinobacter, Pseudomonas, Thalassospira, and Vibrio. Pseudomonas aeruginosa contained the genes (napAB, norBC, nirS, and nosZ) for complete denitrification and may be responsible for mediating denitrification. 66% representative isolates (16/24) contained genes for reducing nitrate to nitrite (nasA, napAB, or narGHI) and 79% representative isolates (19/24) possessed genes for converting nitrite to ammonia (nirA or nirBD), suggesting that nitrate and nitrite could act as electron acceptors to generate ammonium, subsequently being utilized as a reduced nitrogen source. This study improves our understanding of the microbial diversity within the OMZ of Changjiang Estuary and may facilitate the cultivation and exploitation of bacteria involved in the nitrogen cycle.

To better understand bacterial communities and metabolism under nitrogen deficiency, 154 seawater samples were obtained from 0-200 m at 22 stations in the photic zone of the Western North Pacific Ocean and, from them, 634 nitrate-utilizing bacteria were isolated using selective media and culture-dependent methods. Twenty-nine of 94 representative nitrate-utilizing isolates were found to be nitrate reducers, among which Qipengyuania flava, Roseibium aggregatum, Erythrobacter aureus, Vibrio campbellii, and Stappia indica were identified from all tested seawater layers of the photic zone and at almost all stations. The presence of multiple genes for chitin degradation and extracellular peptidases may indicate that almost all nitrate-reducing isolates (28/29) can use chitin and proteinaceous compounds as important sources of carbon and nitrogen. Most strains contained assimilatory nitrate reduction or dissimilatory nitrate reduction to ammonia genes suggesting that nitrate and nitrite could act as electron acceptors to generate ammonia. All of the nitrate-reducing isolates contained genes encoding the assimilatory sulfate reduction pathway, and six of the nitrate-reducing isolates encoded sulfur oxidization genes, indicating that they could oxidize sulfur. Five nitrate-reducing isolates obtained near the chlorophyll a-maximum layer contained a dimethylsulfoniopropionate synthesis gene and three of them contained both dimethylsulfoniopropionate synthesis and cleavage genes. This suggests that nitrate-reducing isolates may participate in dimethylsulfoniopropionate synthesis and catabolism in photic seawater. Collectively, these results reveal nitrate-reducing bacterial diversity and have implications for understanding the role of such strains in the ecology and biogeochemical cycles of nitrogen, carbon, and sulfur in the oligotrophic marine photic zone.

To better understand bacterial communities and metabolism under nitrogen deficiency, 154 seawater samples were obtained from 5 to 200 m at 22 stations in the photic zone of the Western North Pacific Ocean. Total 634 nitrate-utilizing bacteria were isolated using selective media and culture-dependent methods, and 295 of them were positive for nitrate reduction. These nitrate-reducing bacteria belonged to 19 genera and 29 species and among them, Qipengyuania flava, Roseibium aggregatum, Erythrobacter aureus, Vibrio campbellii, and Stappia indica were identified from all tested seawater layers of the photic zone and at almost all stations. Twenty-nine nitrate-reducing strains representing different species were selected for further the study of nitrogen, sulfur, and carbon metabolism. All 29 nitrate-reducing isolates contained genes encoding dissimilatory nitrate reduction or assimilatory nitrate reduction. Six nitrate-reducing isolates can oxidize thiosulfate based on genomic analysis and activity testing, indicating that nitrate-reducing thiosulfate-oxidizing bacteria exist in the photic zone. Five nitrate-reducing isolates obtained near the chlorophyll a-maximum layer contained a dimethylsulfoniopropionate synthesis gene and three of them contained both dimethylsulfoniopropionate synthesis and cleavage genes. This suggests that nitrate-reducing isolates may participate in dimethylsulfoniopropionate synthesis and catabolism in photic seawater. The presence of multiple genes for chitin degradation and extracellular peptidases may indicate that almost all nitrate-reducing isolates (28/29) can use chitin and proteinaceous compounds as important sources of carbon and nitrogen. Collectively, these results reveal culturable nitrate-reducing bacterial diversity and have implications for understanding the role of such strains in the ecology and biogeochemical cycles of nitrogen, sulfur, and carbon in the oligotrophic marine photic zone.