Phosphate diester utilization by marine diazotrophs Trichodesmium erythraeum and Crocosphaera watsonii

In the phosphate-depleted oligotrophic ocean, microbes utilize various dissolved organic phosphorus (P) compounds as alternative P sources, using enzymes such as alkaline phosphatases. However, knowledge of such P acquisition mechanisms is limited, especially in association with the physiology of nitrogen-fixing organisms, which play a substantial role in marine biogeochemical cycling. We show that nonaxenic clonal cultures of 2 oceanic diazotrophs, Trichodesmium erythraeum and Crocosphaera watsonii, have the ability to utilize phosphate diester as their sole P source, using a model artificial compound—bis-p-nitrophenyl phosphate (bisNPP). Although both diazotroph cultures likely preferred phosphate monoester to diester, the expressed diesterase activity was theoretically sufficient to fulfill their P demands, and they showed significant growth in bisNPP-added media. Interestingly, a distinct difference in their growth trends was observed, with faster onset of growth by C. watsonii and delayed onset of growth by T. erythraeum. This indicates that the C. watsonii consortium can effectively and rapidly assimilate in situ diesters as alternative P sources in the field. Nonetheless, when considering the poor bisNPP utilization reported from other marine phytoplankton taxa, our results indicate that the utilization of particular diester compounds is a notable and advantageous strategy for both diazotroph consortia to alleviate P limitation in the oligotrophic ocean.

Transfer of diazotroph-derived nitrogen towards non-diazotrophic planktonic communities: a comparative study between <i>Trichodesmium</i> <i>erythraeum</i>, <i>Crocosphaera watsonii</i> and <i>Cyanothece</i> sp.

Biological dinitrogen (N2) fixation is the major source of new nitrogen (N) for the open ocean, and thus promotes marine productivity, in particular in the vast N-depleted regions of the surface ocean. Yet, the fate of the diazotroph-derived N (DDN) in marine ecosystems is poorly understood, and its transfer to auto- and heterotrophic surrounding plankton communities is rarely measured due to technical limitations. Moreover, the different diazotrophs involved in N2 fixation (Trichodesmium spp. vs. UCYN) exhibit distinct patterns of N2 fixation and inhabit different ecological niches, thus having potentially different fates in the marine food webs that remain to be explored. Here we used nanometer scale secondary ion mass spectrometry (nanoSIMS) coupled with 15N2 isotopic labelling and flow cytometry cell sorting to examine the DDN transfer to specific groups of natural phytoplankton and bacteria during artificially induced diazotroph blooms in New Caledonia (southwestern Pacific). The fate of the DDN was compared according to the three diazotrophs: the filamentous and colony-forming Trichodesmium erythraeum (IMS101), and the unicellular strains Crocosphaera watsonii WH8501 and Cyanothece ATCC51142. After 48 h, 7–17 % of the N2 fixed during the experiment was transferred to the dissolved pool and 6–12 % was transferred to non-diazotrophic plankton. The transfer was twice as high in the T. erythraeum bloom than in the C. watsonii and Cyanothece blooms, which shows that filamentous diazotrophs blooms are more efficient at promoting non-diazotrophic production in N-depleted areas. The amount of DDN released in the dissolved pool did not appear to be a good indicator of the DDN transfer efficiency towards the non-diazotrophic plankton. In contrast, the 15N-enrichment of the extracellular ammonium (NH4+) pool was a good indicator of the DDN transfer efficiency: it was significantly higher in the T. erythraeum than in unicellular diazotroph blooms, leading to a DDN transfer twice as efficient. This suggests that NH4+ was the main pathway of the DDN transfer from diazotrophs to non-diazotrophs. The three simulated diazotroph blooms led to significant increases in non-diazotrophic plankton biomass. This increase in biomass was first associated with heterotrophic bacteria followed by phytoplankton, indicating that heterotrophs took the most advantage of the DDN in this oligotrophic ecosystem.

Transfer of diazotroph derived nitrogen towards non-diazotrophic planktonic communities: a comparative study between <i>Trichodesmium</i> <i>erythraeum</i>, <i>Crocosphaera watsonii</i> and <i>Cyanothece</i> sp.

Biological dinitrogen (N2) fixation is the major source of new nitrogen (N) for the open ocean, and thus promotes marine productivity, in particular in the vast N-depleted regions of the surface ocean. Yet, the fate of the diazotroph-derived N (DDN) in marine ecosystems is poorly understood and its transfer to auto- and heterotrophic surrounding plankton communities is rarely measured due to technical limitations. Moreover, the different diazotrophs involved in N2 fixation (Trichodesmium spp. vs. UCYN) exhibit distinct patterns of N2 fixation and inhabit different ecological niches, thus having potentially different fates in the marine food webs, that remains to be explored. Here we used nanometer scale secondary ion mass spectrometry (nanoSIMS) coupled with 15N2 isotopic labelling and flow cytometry cell sorting to examine the DDN transfer to specific groups of natural phytoplankton and bacteria during artificially-induced diazotroph blooms in New Caledonia (southwestern Pacific). The fate of the DDN was compared according to the three diazotrophs: the filamentous and colony forming Trichodesmium erythraeum (IMS101), and the unicellular strains Crocosphaera watsonii WH8501 and Cyanothece ATCC51142. After 48 h, 7–17 % of the N2 fixed during the experiment was transferred to the dissolved pool and 6–12 % was transferred to non-diazotrophic plankton. The transfer was twice as high during the T. erythraeum bloom than during the C. watsonii and Cyanothece blooms, arguing that filamentous diazotrophs blooms are more efficient at promoting non-diazotrophic production in N depleted areas. The amount of DDN released in the dissolved pool did not appear as a good indicator of the DDN transfer efficiency towards the non-diazotrophic plankton. In contrast, the 15N-enrichment of the extracellular ammonium (NH4+) pool was a good indicator of the DDN transfer efficiency: it was significantly higher in the T. erythraeum than in unicellular diazotroph blooms, leading to a DDN transfer twice as efficient. This suggests that NH4+ was the main pathway of the DDN transfer from diazotrophs to non-diazotrophs. The three simulated diazotroph blooms led to significant increases in non-diazotrophic plankton biomass. This increase in biomass was first associated with heterotrophic bacteria followed phytoplankton, indicating that heterotrophs took the most advantage of the DDN in this oligotrophic ecosystem.