Specific hopanoid classes differentially affect free-living and symbiotic states of Bradyrhizobium diazoefficiens

Gargi Kulkarni (1) , Nicolas Busset (2) , Antonio Molinaro (3) , Daniel Gargani (4, 5) , Clémence Chaintreuil (2) , Alba Silipo (3) , Eric Giraud (2) , Dianne K. Newman (1, 6, 7)

1 Division of Biology and Biological Engineering
2 UMR LSTM - Laboratoire des symbioses tropicales et méditerranéennes
3 Dipartimento di Scienze Chimiche
4 UMR BGPI - Biologie et Génétique des Interactions Plante-Parasite
5 Cirad - Centre de Coopération Internationale en Recherche Agronomique pour le Développement
6 HHMI - Howard Hughes Medical Institute
7 Division of Geological and Planetary Sciences [Pasadena]

A better understanding of how bacteria resist stresses encountered during the progression of plant-microbe symbioses will advance our ability to stimulate plant growth. Here, we show that the symbiotic system comprising the nitrogen-fixing bacterium Bradyrhizobium diazoefficiens and the legume Aeschynomene afraspera requires hopanoid production for optimal fitness. While methylated (2Me) hopanoids contribute to growth under plant-cell-like microaerobic and acidic conditions in the free-living state, they are dispensable during symbiosis. In contrast, synthesis of extended (C-35) hopanoids is required for growth microaerobically and under various stress conditions (high temperature, low pH, high osmolarity, bile salts, oxidative stress, and antimicrobial peptides) in the free-living state and also during symbiosis. These defects might be due to a less rigid membrane resulting from the absence of free or lipidA-bound C-35 hopanoids or the accumulation of the C-35 hopanoid diploptene. Our results also show that C-35 hopanoids are necessary for symbiosis only with the host Aeschynomene afraspera but not with soybean. This difference is likely related to the presence of cysteine-rich antimicrobial peptides in Aeschynomene nodules that induce drastic modification in bacterial morphology and physiology. The study of hopanoid mutants in plant symbionts thus provides an opportunity to gain insight into host-microbe interactions during later stages of symbiotic progression, as well as the microenvironmental conditions for which hopanoids provide a fitness advantage. IMPORTANCE Because bradyrhizobia provide fixed nitrogen to plants, this work has potential agronomical implications. An understanding of how hopanoids facilitate bacterial survival in soils and plant hosts may aid the engineering of more robust agronomic strains, especially relevant in regions that are becoming warmer and saline due to climate change. Moreover, this work has geobiological relevance: hopanes, molecular fossils of hopanoids, are enriched in ancient sedimentary rocks at discrete intervals in Earth history. This is the first study to uncover roles for 2Me- and C-35 hopanoids in the context of an ecological niche that captures many of the stressful environmental conditions thought to be important during (2Me)-hopane deposition. Though much remains to be done to determine whether the conditions present within the plant host are shared with niches of relevance to the rock record, our findings represent an important step toward identifying conserved mechanisms whereby hopanoids contribute to fitness.

Mots clés: symbiosis; plant - microorganism relationship

Mots clés: bradyrhizobium; croissance des plantes; aeschynomene; symbiose; relation plante-microorganisme; écologie de la rhizosphère; fixation de l'azote

Domaines: Biodiversité et Ecologie

Gargi Kulkarni, Nicolas Busset, Antonio Molinaro, Daniel Gargani, Clémence Chaintreuil, et al.. Specific hopanoid classes differentially affect free-living and symbiotic states of Bradyrhizobium diazoefficiens. mBio, 2015, 6 (5), 9 p. ⟨10.1128/mBio.01251-15⟩. ⟨hal-01594981⟩