Abstract

In the most studied rhizobium-legume interactions, the host plant supplies the symbiont with homocitrate, an essential co-factor of the nitrogenase enzyme complex, via the expression of a nodule-specific homocitrate synthase FEN1. Photosynthetic bradyrhizobia interacting with Nod factor (NF) dependent and NF-independent Aeschynomene legumes are able to synthesize homocitrate themselves as they contain a nifV gene encoding a homocitrate synthase. Here, we show that in the model strain ORS285, nifV is required for free-living and symbiotic dinitrogen fixation with NF-independent Aeschynomene species. In contrast, in symbiosis with NF-dependent Aeschynomene species, the nifV requirement for efficient nitrogen fixation was found to be host plant dependent. Interestingly, orthologs of FEN1 were found in both NF-dependent and NF-independent Aeschynomene species. However, a high nodule specific induction of FEN1 expression was only observed in A. afraspera, a host plant in which nifV is not required for symbiotic dinitrogen fixation. These data indicate that efficient symbiotic nitrogen fixation in many of the tested Aeschynomene species requires rhizobial homocitrate synthesis. Considering that more than 10% of the fully sequenced rhizobium strains do contain a nifV gene, the Aeschynomene/photosynthetic Bradyrhizobium interaction is likely not the only rhizobium/legume symbiosis where rhizobial nifV expression is required.

Highlights

  • Nitrogen is an essential element for all living organisms

  • It has been demonstrated that Lotus japonicus expresses a nodule specific homocitrate synthase (FEN1) that compensates for the absence of homocitrate synthase activity in the bacterial partner Mesorhizobium loti[5]

  • Azorhizobium caulinodans ORS571 and photosynthetic Bradyrhizobium strains are examples of rhizobia that contain a nifV gene. As these rhizobia are capable to fix dinitrogen under free-living conditions[6, 7], we can ask if the presence of the nifV gene in these rhizobia is related to this capacity or because their host plant are unable to supply homocitrate during symbiosis

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Summary

Introduction

Nitrogen is an essential element for all living organisms. On earth the major source of nitrogen is atmospheric dinitrogen, which is fixed by microorganism (diazotrophs) that are able to reduce dinitrogen to ammonium by a nitrogenase enzyme complex. Azorhizobium caulinodans ORS571 and photosynthetic Bradyrhizobium strains are examples of rhizobia that contain a nifV gene As these rhizobia are capable to fix dinitrogen under free-living conditions[6, 7], we can ask if the presence of the nifV gene in these rhizobia is related to this capacity or because their host plant are unable to supply homocitrate during symbiosis. To investigate this question, we have constructed a nifV deletion mutant in the photosynthetic Bradyrhizobium strain ORS285. We correlated the nodulation phenotypes with the presence and expression of FEN1 orthologs in A. afraspera and A. evenia, representatives of the NF-dependent and NF-independent Aeschynomene groups, respectively

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