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Sustainable-forage production is globally increasing, especially in marginal areas where the edaphic conditions for plant growth are not optimal. Soil salinization influences the symbiotic interaction between alfalfa and rhizobia. The efficiency of different symbiotic pairs (Sinorhizobium meliloti—Medicago sativa) was evaluated in relation to NaCl application (100 mM) on two different alfalfa cultivars (Marina and Etrusca) and 21 S. meliloti strains isolated in Algeria. At 100 mM NaCl, it was observed that there was a higher variability of plant dry weight compared to the control. The strains able to improve plant growth at 100 mM NaCl were different and specific for each alfalfa cultivar, highlighting that (symbiont) G × (host) G interaction is magnified under stressed (saline) conditions (E). Three strains were then identified as candidate inoculants for M. sativa cv Marina and used for an in-field experiment with induced stress (no irrigation), together with S. meliloti GR4 (a highly competitive strain). In-field experiments, showed a high variability, and a significant difference of plant biomass was observed only for those inoculated with S. meliloti GR4. Obtained results suggest that multiple traits should be considered for inoculant-strain selection, and for an efficient translation from lab to field, it requires extensive comprehension of the mechanisms driving G × G × E interaction.

Sustainable-forage production is globally increasing, especially in marginal areas where the edaphic conditions for plant growth are not optimal. Soil salinization influences the symbiotic interaction between alfalfa and rhizobia. The efficiency of different symbiotic pairs (Sinorhizobium meliloti—Medicago sativa) was evaluated in relation to NaCl application (100 mM) on two different alfalfa cultivars (Marina and Etrusca) and 21 S. meliloti strains isolated in Algeria. At 100 mM NaCl, it was observed that there was a higher variability of plant dry weight compared to the control. The strains able to improve plant growth at 100 mM NaCl were different and specific for each alfalfa cultivar, highlighting that (symbiont) G × (host) G interaction is magnified under stressed (saline) conditions (E). Three strains were then identified as candidate inoculants for M. sativa cv Marina and used for an in-field experiment with induced stress (no irrigation), together with S. meliloti GR4 (a highly competitive strain). In-field experiments, showed a high variability, and a significant difference of plant biomass was observed only for those inoculated with S. meliloti GR4. Obtained results suggest that multiple traits should be considered for inoculant-strain selection, and for an efficient translation from lab to field, it requires extensive comprehension of the mechanisms driving G × G × E interaction.

Sustainable-forage production is globally increasing, especially in marginal areas where the edaphic conditions for plant growth are not optimal. Soil salinization influences the symbiotic interaction between alfalfa and rhizobia. The efficiency of different symbiotic pairs (Sinorhizobium meliloti—Medicago sativa) was evaluated in relation to NaCl application (100 mM) on two different alfalfa cultivars (Marina and Etrusca) and 21 S. meliloti strains isolated in Algeria. At 100 mM NaCl, it was observed that there was a higher variability of plant dry weight compared to the control. The strains able to improve plant growth at 100 mM NaCl were different and specific for each alfalfa cultivar, highlighting that (symbiont) G × (host) G interaction is magnified under stressed (saline) conditions (E). Three strains were then identified as candidate inoculants for M. sativa cv Marina and used for an in-field experiment with induced stress (no irrigation), together with S. meliloti GR4 (a highly competitive strain). In-field experiments, showed a high variability, and a significant difference of plant biomass was observed only for those inoculated with S. meliloti GR4. Obtained results suggest that multiple traits should be considered for inoculant-strain selection, and for an efficient translation from lab to field, it requires extensive comprehension of the mechanisms driving G × G × E interaction.

Sustainable-forage production is globally increasing, especially in marginal areas where the edaphic conditions for plant growth are not optimal. Soil salinization influences the symbiotic interaction between alfalfa and rhizobia. The efficiency of different symbiotic pairs (Sinorhizobium meliloti—Medicago sativa) was evaluated in relation to NaCl application (100 mM) on two different alfalfa cultivars (Marina and Etrusca) and 21 S. meliloti strains isolated in Algeria. At 100 mM NaCl, it was observed that there was a higher variability of plant dry weight compared to the control. The strains able to improve plant growth at 100 mM NaCl were different and specific for each alfalfa cultivar, highlighting that (symbiont) G × (host) G interaction is magnified under stressed (saline) conditions (E). Three strains were then identified as candidate inoculants for M. sativa cv Marina and used for an in-field experiment with induced stress (no irrigation), together with S. meliloti GR4 (a highly competitive strain). In-field experiments, showed a high variability, and a significant difference of plant biomass was observed only for those inoculated with S. meliloti GR4. Obtained results suggest that multiple traits should be considered for inoculant-strain selection, and for an efficient translation from lab to field, it requires extensive comprehension of the mechanisms driving G × G × E interaction.

Methylation of specific DNA sequences is ubiquitous in bacteria and has known roles in immunity and regulation of cellular processes, such as the cell cycle. Here, we explored DNA methylation in bacteria of the genus Ensifer, including its potential role in regulating terminal differentiation during nitrogen-fixing symbiosis with legumes. Using single-molecule real-time sequencing, six genome-wide methylated motifs were identified across four Ensifer strains, five of which were strain-specific. Only the GANTC motif, recognized by the cell cycle-regulated CcrM methyltransferase, was methylated in all strains. In actively dividing cell cultures, methylation of GANTC motifs increased progressively from the ori to ter regions in each replicon, in agreement with a cell cycle-dependent regulation of CcrM. In contrast, there was near full genome-wide GANTC methylation in the early stage of symbiotic differentiation. This was followed by a moderate decrease in the overall extent of methylation and a progressive decrease in chromosomal GANTC methylation from the ori to ter regions in later stages of differentiation. Based on these observations, we suggest that CcrM activity is dysregulated and constitutive during terminal differentiation, which we hypothesize is a driving factor for endoreduplication of terminally differentiated bacteroids. IMPORTANCE Nitrogen fixation by rhizobia in symbiosis with legumes is economically and ecologically important. The symbiosis can involve a complex bacterial transformation-terminal differentiation-that includes major shifts in the transcriptome and cell cycle. Epigenetic regulation is an important regulatory mechanism in diverse bacteria; however, the roles of DNA methylation in rhizobia and symbiotic nitrogen fixation have been poorly investigated. We show that aside from cell cycle regulation, DNA methyltransferases are unlikely to have conserved roles in the biology of bacteria of the genus Ensifer. However, we present evidence consist

Methylation of specific DNA sequences is ubiquitous in bacteria and has known roles in immunity and regulation of cellular processes, such as the cell cycle. Here, we explored DNA methylation in bacteria of the genus Ensifer, including its potential role in regulating terminal differentiation during nitrogen-fixing symbiosis with legumes. Using single-molecule real-time sequencing, six genome-wide methylated motifs were identified across four Ensifer strains, five of which were strain-specific. Only the GANTC motif, recognized by the cell cycle-regulated CcrM methyltransferase, was methylated in all strains. In actively dividing cell cultures, methylation of GANTC motifs increased progressively from the ori to ter regions in each replicon, in agreement with a cell cycle-dependent regulation of CcrM. In contrast, there was near full genome-wide GANTC methylation in the early stage of symbiotic differentiation. This was followed by a moderate decrease in the overall extent of methylation and a progressive decrease in chromosomal GANTC methylation from the ori to ter regions in later stages of differentiation. Based on these observations, we suggest that CcrM activity is dysregulated and constitutive during terminal differentiation, which we hypothesize is a driving factor for endoreduplication of terminally differentiated bacteroids. IMPORTANCE Nitrogen fixation by rhizobia in symbiosis with legumes is economically and ecologically important. The symbiosis can involve a complex bacterial transformation-terminal differentiation-that includes major shifts in the transcriptome and cell cycle. Epigenetic regulation is an important regulatory mechanism in diverse bacteria; however, the roles of DNA methylation in rhizobia and symbiotic nitrogen fixation have been poorly investigated. We show that aside from cell cycle regulation, DNA methyltransferases are unlikely to have conserved roles in the biology of bacteria of the genus Ensifer. However, we present evidence consist

Many molecular signals are exchanged between rhizobia and host legume plants, some of which are crucial for symbiosis to take place, while others are modifiers of the interaction, which have great importance in the competition with the soil microbiota and in the genotype-specific perception of host plants. Here, we review recent findings on strain-specific and host genotype-specific interactions between rhizobia and legumes, discussing the molecular actors (genes, gene products and metabolites) which play a role in the establishment of symbiosis, and highlighting the need for research including the other components of the soil (micro)biota, which could be crucial in developing rational-based strategies for bioinoculants and synthetic communities’ assemblage.

Plant-associated microbiota are becoming central in the development of ways to improve plant productivity and health. However, most research has focussed mainly on a few model plant species. It is essential to translate discoveries to the many nonmodel crops, allowing the design and application of effective synthetic microbiota.

Many molecular signals are exchanged between rhizobia and host legume plants, some of which are crucial for symbiosis to take place, while others are modifiers of the interaction, which have great importance in the competition with the soil microbiota and in the genotype-specific perception of host plants. Here, we review recent findings on strain-specific and host genotype-specific interactions between rhizobia and legumes, discussing the molecular actors (genes, gene products and metabolites) which play a role in the establishment of symbiosis, and highlighting the need for research including the other components of the soil (micro)biota, which could be crucial in developing rational-based strategies for bioinoculants and synthetic communities’ assemblage.

Plant-associated microbiota are becoming central in the development of ways to improve plant productivity and health. However, most research has focussed mainly on a few model plant species. It is essential to translate discoveries to the many nonmodel crops, allowing the design and application of effective synthetic microbiota.

Many molecular signals are exchanged between rhizobia and host legume plants, some of which are crucial for symbiosis to take place, while others are modifiers of the interaction, which have great importance in the competition with the soil microbiota and in the genotype-specific perception of host plants. Here, we review recent findings on strain-specific and host genotype-specific interactions between rhizobia and legumes, discussing the molecular actors (genes, gene products and metabolites) which play a role in the establishment of symbiosis, and highlighting the need for research including the other components of the soil (micro)biota, which could be crucial in developing rational-based strategies for bioinoculants and synthetic communities’ assemblage.

The rhizosphere and rhizoplane are nutrient-rich but selective environments for the root microbiome. Here, we deciphered a posttranscriptional network regulated by the homologous trans-small RNAs (sRNAs) AbcR1 and AbcR2, which rewire the metabolism of the nitrogen-fixing a-rhizobium Sinorhizobium meliloti during preinfection stages of symbiosis with its legume host alfalfa. The LysR-type regulator LsrB, which transduces the cell redox state, is indispensable for AbcR1 expression in actively dividing bacteria, whereas the stress-induced transcription of AbcR2 depends on the alternative s factor RpoH1. MS2 affinity purification coupled with RNA sequencing unveiled exceptionally large and overlapping AbcR1/2 mRNA interactomes, jointly representing;6% of the S. meliloti protein-coding genes. Most mRNAs encode transport/metabolic proteins whose translation is silenced by base pairing to two distinct anti-Shine Dalgarno motifs that function independently in both sRNAs. A metabolic model-aided analysis of the targetomes predicted changes in AbcR1/2 expression driven by shifts in carbon/nitrogen sources, which were confirmed experimentally. Low AbcR1/2 levels in some defined media anticipated overexpression growth phenotypes linked to the silencing of specific mRNAs. As a proof of principle, we confirmed AbcR1/2-mediated downregulation of the L-amino acid AapQ permease. AbcR1/2 interactomes are well represented in rhizosphere-related S. meliloti transcriptomic signatures. Remarkably, a lack of AbcR1 specifically compromised the ability of S. meliloti to colonize the root rhizoplane. The AbcR1 regulon likely ranks the utilization of available substrates to optimize metabolism, thus conferring on S. meliloti an advantage for efficient rhizosphere/rhizoplane colonization. AbcR1 regulation is predicted to be conserved in related a-rhizobia, which opens unprecedented possibilities for engineering highly competitive biofertilizers. IMPORTANCE Nitrogen-fixing root nodule symbioses

Many molecular signals are exchanged between rhizobia and host legume plants, some of which are crucial for symbiosis to take place, while others are modifiers of the interaction, which have great importance in the competition with the soil microbiota and in the genotype-specific perception of host plants. Here, we review recent findings on strain-specific and host genotype-specific interactions between rhizobia and legumes, discussing the molecular actors (genes, gene products and metabolites) which play a role in the establishment of symbiosis, and highlighting the need for research including the other components of the soil (micro)biota, which could be crucial in developing rational-based strategies for bioinoculants and synthetic communities’ assemblage.

Plant-associated microbiota are becoming central in the development of ways to improve plant productivity and health. However, most research has focussed mainly on a few model plant species. It is essential to translate discoveries to the many nonmodel crops, allowing the design and application of effective synthetic microbiota.

Plant-associated microbiota are becoming central in the development of ways to improve plant productivity and health. However, most research has focussed mainly on a few model plant species. It is essential to translate discoveries to the many nonmodel crops, allowing the design and application of effective synthetic microbiota.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

Bacterial endophytes support the adaptation of host plants to harsh environments. In this study, culturable bacterial endophytes were isolated from the African rice Oryza glaberrima L., which is well-adapted to grow with poor external inputs in the tropical region of Mali. Among these, six N-fixer strains were used to inoculate O. glaberrima RAM133 and the Asian rice O. sativa L. cv. Baldo, selected for growth in temperate climates. The colonization efficiency and the N-fixing activity were evaluated and compared for the two rice varieties. Oryza sativa-inoculated plants showed a fairly good colonization efficiency and nitrogenase activity. The inoculation of Oryza sativa with the strains Klebsiella pasteurii BDA134-6 and Phytobacter diazotrophicus BDA59-3 led to the highest nitrogenase activity. In addition, the inoculation of ‘Baldo’ plants with the strain P. diazotrophicus BDA59-3 led to a significant increase in nitrogen, carbon and chlorophyll content. Finally, ‘Baldo’ plants inoculated with Kl. pasteurii BDA134-6 showed the induction of antioxidant enzymes activity and the maintenance of nitrogen-fixation under salt stress as compared to the unstressed controls. As these endophytes efficiently colonize high-yielding crop varieties grown in cold temperate climates, they become good candidates to promote their growth under unfavorable conditions.

In the past 10 years, the average demand for meat and milk across the world has significantly increased, especially in developing countries. Therefore, to support the production of animal-derived food products, a huge quantity of feed resources is needed. This paper does not present original research, but rather provides a conceptual strategy to improve primary production in a sustainable way, in relation to forthcoming issues linked to climate change. Increases in meat and milk production could be achieved by formulating balanced diets for ovines based on alfalfa integrated with local agricultural by-products. As the central component of the diet is alfalfa, one goal of the project is increasing the yield of alfalfa in a sustainable way via inoculating seeds with symbiotic rhizobia (i.e., Sinorhizobium meliloti). Seed inoculants are already present on the market but have not been optimized for arid soils. Furthermore, a part of the project is focused on the selection of elite symbiotic strains that show increased resistance to salt stress and competitiveness. The second component of the experimental diets is bio-waste, especially that obtained from olive oil manufacturing (i.e., pomace). The addition of agro-by-products allows us to use such waste as a resource for animal feeding, and possibly, to modulate rumen metabolism, thereby increasing the nutritional quality of milk and meat.

Bacterial genome evolution is characterized by gains, losses, and rearrangements of functional genetic segments. The extent to which large-scale genomic alterations influence genotype-phenotype relationships has not been investigated in a high-throughput manner. In the symbiotic soil bacterium Sinorhizobium meliloti, the genome is composed of a chromosome and two large extrachromosomal replicons (pSymA and pSymB, which together constitute 45% of the genome). Massively parallel transposon insertion sequencing (Tn-seq) was employed to evaluate the contributions of chromosomal genes to growth fitness in both the presence and absence of these extrachromosomal replicons. Ten percent of chromosomal genes from diverse functional categories are shown to genetically interact with pSymA and pSymB. These results demonstrate the pervasive robustness provided by the extrachromosomal replicons, which is further supported by constraint-based metabolic modeling. A comprehensive picture of core S. meliloti metabolism was generated through a Tn-seq-guided in silico metabolic network reconstruction, producing a core network encompassing 726 genes. This integrated approach facilitated functional assignments for previously uncharacterized genes, while also revealing that Tn-seq alone missed over a quarter of wild-type metabolism. This work highlights the many functional dependencies and epistatic relationships that may arise between bacterial replicons and across a genome, while also demonstrating how Tn-seq and metabolic modeling can be used together to yield insights not obtainable by either method alone.

In all domains of life, proper regulation of the cell cycle is critical to coordinate genome replication, segregation and cell division. In some groups of bacteria, e.g. Alphaproteobacteria, tight regulation of the cell cycle is also necessary for the morphological and functional differentiation of cells. Sinorhizobium meliloti is an alphaproteobacterium that forms an economically and ecologically important nitrogen-fixing symbiosis with specific legume hosts. During this symbiosis S. meliloti undergoes an elaborate cellular differentiation within host root cells. The differentiation of S. meliloti results in massive amplification of the genome, cell branching and/or elongation, and loss of reproductive capacity. In Caulobacter crescentus, cellular differentiation is tightly linked to the cell cycle via the activity of the master regulator CtrA, and recent research in S. meliloti suggests that CtrA might also be key to cellular differentiation during symbiosis. However, the regulatory circuit driving cell cycle progression in S. meliloti is not well characterized in both the free-living and symbiotic state. Here, we investigated the regulation and function of CtrA in S. meliloti. We demonstrated that depletion of CtrA cause cell elongation, branching and genome amplification, similar to that observed in nitrogen-fixing bacteroids. We also showed that the cell cycle regulated proteolytic degradation of CtrA is essential in S. meliloti, suggesting a possible mechanism of CtrA depletion in differentiated bacteroids. Using a combination of ChIP-Seq and gene expression microarray analysis we found that although S. meliloti CtrA regulates similar processes as C. crescentus CtrA, it does so through different target genes. For example, our data suggest that CtrA does not control the expression of the Fts complex to control the timing of cell division during the cell cycle, but instead it negatively regulates the septum-inhibiting Min system. Our findings provide valuable in, France. Agence nationale de la recherche (ANR-JCJC-2011-Castacc), National Institutes of Health (U.S.) (Grant GM31010), National Institutes of Health (U.S.) (Grant P30 ES002109), National Institutes of Health (U.S.) (Pre-doctoral Training Grant T32GM007287)

As is well known, pathogenic microbes evolve rapidly to escape from the host immune system and antibiotics. Genetic variations among microbial populations occur frequently during the long-term pathogen–host evolutionary arms race, and individual mutation beneficial for the fitness can be fixed preferentially. Many recent comparative genomics studies have pointed out the importance of selective forces in the molecular evolution of bacterial pathogens. The public availability of large-scale next-generation sequencing data and many state-of-the-art statistical methods of molecular evolution enable us to scan genome-wide alignments for evidence of positive Darwinian selection, recombination, and other evolutionary forces operating on the coding regions. In this chapter, we describe an integrative analysis pipeline and its application to tracking featured evolutionary trajectories on the genome of an animal pathogen. The evolutionary analysis of the protein-coding part of the genomes will provide a wide spectrum oof genetic variations that play potential roles in adaptive evolution of bacteria.

In this work we analyzed the composition and structure of cultivable bacterial communities isolated from the stem/leaf and root compartments of two medicinal plants, Echinacea purpurea (L.) Moench and Echinacea angustifolia (DC.) Hell, grown in the same soil, as well as the bacterial community from their rhizospheric soils. Molecular PCR-based techniques were applied to cultivable bacteria isolated from the three compartments of the two plants. The results showed that the two plants and their respective compartments were characterized by different communities, indicating a low degree of strain sharing and a strong selective pressure within plant tissues. Pseudomonas was the most highly represented genus, together with Actinobacteria and Bacillus spp. The presence of distinct bacterial communities in different plant species and among compartments of the same plant species could account for the differences in the medicinal properties of the two plants. [Int Microbiol 2014; 17(3):165-174]Keywords: Echinacea purpurea · Echinacea angustifolia · rhizosphere · medicinal plants · endophytes

Sinorhizobium meliloti is a soil bacterium that invades the root nodules it induces on Medicago sativa, whereupon it undergoes an alteration of its cell cycle and differentiates into nitrogen-fixing, elongated and polyploid bacteroid with higher membrane permeability. In Caulobacter crescentus, a related alphaproteobacterium, the principal cell cycle regulator, CtrA, is inhibited by the phosphorylated response regulator DivK. The phosphorylation of DivK depends on the histidine kinase DivJ, while PleC is the principal phosphatase for DivK. Despite the importance of the DivJ in C. crescentus, the mechanistic role of this kinase has never been elucidated in other Alphaproteobacteria. We show here that the histidine kinases DivJ together with CbrA and PleC participate in a complex phosphorylation system of the essential response regulator DivK in S. meliloti. In particular, DivJ and CbrA are involved in DivK phosphorylation and in turn CtrA inactivation, thereby controlling correct cell cycle progression and the integrity of the cell envelope. In contrast, the essential PleC presumably acts as a phosphatase of DivK. Interestingly, we found that a DivJ mutant is able to elicit nodules and enter plant cells, but fails to establish an effective symbiosis suggesting that proper envelope and/or low CtrA levels are required for symbiosis., National Institutes of Health (U.S.) (Grant GM31010)

In this work we analyzed the composition and structure of cultivable bacterial communities isolated from the stem/leaf and root compartments of two medicinal plants, Echinacea purpurea (L.) Moench and Echinacea angustifolia (DC.) Hell, grown in the same soil, as well as the bacterial community from their rhizospheric soils. Molecular PCR-based techniques were applied to cultivable bacteria isolated from the three compartments of the two plants. The results showed that the two plants and their respective compartments were characterized by different communities, indicating a low degree of strain sharing and a strong selective pressure within plant tissues. Pseudomonas was the most highly represented genus, together with Actinobacteria and Bacillus spp. The presence of distinct bacterial communities in different plant species and among compartments of the same plant species could account for the differences in the medicinal properties of the two plants. [Int Microbiol 2014; 17(3):165-174]Keywords: Echinacea purpurea · Echinacea angustifolia · rhizosphere · medicinal plants · endophytes

Ensifer (syn. Sinorhizobium) meliloti is an important symbiotic bacterial species that fixes nitrogen. Strains BO21CC and AK58 were previously investigated for their substrate utilization and their plant-growth promoting abilities showing interesting features. Here, we describe the complete genome sequence and annotation of these strains. BO21CC and AK58 genomes are 6,985,065 and 6,974,333 bp long with 6,746 and 6,992 genes predicted, respectively.