Azospirillum brasilense Az39, a model rhizobacterium with AHL quorum quenching capacity

AimsThe aim of this research was to analyze the Quorum sensing (QS) and Quorum quenching (QQ) mechanisms based on N‐acyl‐L‐homoserine lactones (AHLs) in A. brasilense Az39, a strain with remarkable capacity to benefit a wide range of crops under agronomic conditions. Methods and ResultsWe performed an in silico and in vitro analysis of the quorum mechanisms in A. brasilense Az39. The results obtained in vitro using the reporter strains C. violaceum and A. tumefaciens and Liquid Chromatography coupled to Mass‐Mass Spectrometry (LC‐MS/MS) analysis showed that although Az39 does not produce AHL molecules, it is capable of degrading them by at least two hypothetical enzymes identified by bioinformatics approach, associated to the bacterial cell. In Az39 cultures supplemented with 500 nmol l−1 of the C3 unsubstituted AHLs (C4, C6, C8, C10, C12, C14), AHL levels were lower than in non‐inoculated LB media controls. Similar results were observed upon addition of AHLs with hydroxy (OH‐) and keto (oxo‐) substitutions in carbon 3. These results not only demonstrate the ability of Az39 to degrade AHLs. They also show the wide spectrum of molecules that can be degraded by this bacterium. ConclusionsAlthough A.brasilense Az39 is a silent bacterium unable to produce AHL signals, it is able to interrupt the communications between other bacteria and/or plants by a quorum quenching activity

Evaluation of nitrous oxide emission by soybean inoculated with Bradyrhizobium strains commonly used as inoculants in South America

Aims: The purpose of this work was to analyze the agronomic and environmental performance of soybean plants inoculated with the Bradyrhizobium strains widely used as soybean biofertilizers in South America and to determine if these strains possess any functional or taxonomic trait associated with the NO emission. Methods: Bradyrhizobium japonicum E109 and CPAC 15, B. diazoefficiens USDA 110 and CPAC 7, and B. elkanii SEMIA 5019 and SEMIA 587 were used to inoculate soybean seeds. The field experiment was carried out in a soil without history of soybean cultivation in the Argentinian Humid Pampa. The natural N abundance method was applied to estimate N-fixation, and NO production was evaluated using gas chromatography. Among other physiological parameters, shoot dry weight, shoot N content, and crop yield were estimated after harvest. Results: B. japonicum inoculation produced the greatest increases in soybean growth and crop yield but also led to higher NO emissions compared to all other inoculated treatments. Plants inoculated with B. diazoefficiens released the lowest amount of NO, and their growth and yield were the least affected. Inoculation with B. elkanii resulted in intermediate NO emission fluxes and crop yield compared with B. japonicum and B. diazoefficiens. Conclusions: We found that soybean inoculation with strains of B. japonicum and B. elkanii that lack the nosZ gene led to the highest NO emissions under field conditions, but also to the highest crop yield, while inoculation with strains that carry out complete denitrification, nosZ-containing B. diazoefficiens, showed lower NO emission and lower crop yield.To the Instituto de Investigaciones Agrobiotecnológicas (INIAB); Universidad Nacional de Río Cuarto (UNRC); Consejo Nacional de Investigaciones Científcas y Tecnológicas (CONICET), Fondo Nacional de Ciencia y Tecnología (FONCyT); Instituto Nacional de Tecnología Agropecuaria (INTA). FC is Researcher of CONICET at the UNRC. DT and FD are Postdoc and PhD students at the UNRC granted by CONICET. MOC is a former PhD student at the UNRC granted by CONICET. To Mariano Cicchino from INTA Chascomús, who was in charge of sowing and yield estimation at R8. To Juan Pedro Ezquiaga from INTA Castelar, for their contribution to N2O measurements

The importance of denitrification performed by nitrogen-fixing bacteria used as inoculants in South America

Background Replacing synthetic fertilizers by biological nitrogen fixation (BNF) is regarded as an environmentally sound practice, but some diazotrophic bacteria are capable of emitting N2O by denitrification. The ability to use nitrate represents an ecological advantage for the survival of some microorganisms under O2-limiting conditions, but may contribute to increased N2O emissions. Scope The importance of denitrification performed by N2-fixing bacteria used as inoculants in South America is discussed, especially the possibility of these bacteria act as N2O source or sink. Conclusions There is no doubt of the importance of BNF as a sustainable N source for plants. Through genome investigation, we demonstrated that some strains widely used as inoculants for BNF harbor the entire denitrification pathway to reduce nitrate to N2. Others contain none, or only some of the denitrification genes, resulting in complete absence of denitrification or production of intermediates such as NO2−, NO or N2O. Evidence of differential effects of bacterial strains on soil N2O were reported, but more studies are still needed to affirm crop inoculation can be a driver for source or sink of this gas. Finally, considerations were made about BNF as an indispensable resource to indirectly mitigate greenhouse gas emissions in agroecosystems

Azospirillum brasilense Az39, a model rhizobacterium with AHL quorum-quenching capacity

The aim of this research was to analyse the quorum-sensing (QS) and quorum-quenching (QQ) mechanisms based on N-acyl-L-homoserine lactones (AHLs) in Azospirillum brasilense Az39, a strain with remarkable capacity to benefit a wide range of crops under agronomic conditions. We performed an in silico and in vitro analysis of thequorum mechanisms in A. brasilense Az39. The results obtained in vitro using the reporter strains Chromobacterium violaceum and Agrobacterium tumefaciens and liquid chromatography coupled with mass-mass spectrometry analysis showed that although Az39 does not produce AHL molecules, it is capable of degrading them by at least two hypothetical enzymes identified by bioinformatics approach, associated with the bacterial cell. In Az39 cultures supplemented with 500 nmol l-1 of the C3 unsubstituted AHLs (C4, C6, C8, C10, C12, C14), AHL levels were lower than in noninoculated LB media controls. Similar results were observed upon the addition of AHLs with hydroxy (OH-) and keto (oxo-) substitutions in C3. These results not only demonstrate the ability of Az39 to degrade AHLs. They also show the wide spectrum of molecules that can be degraded by this bacterium. Although A. brasilense Az39 is a silent bacterium unable toproduce AHL signals, it is able to interrupt the communications between other bacteria and/or plants by a QQ activity.Significance and Impact of the Study: This is the first report confirming byunequivocal methodology the ability of A. brasilense, one of the most agriculturally used benefic bacteria around the world, to degrade AHLs by a QQ mechanism.Fil: Gualpa, J.. Universidad Nacional de Rio Cuarto. Facultad de Cs.exactas Fisicoquimicas y Naturales. Instituto de Investigaciones Agrobiotecnologicas. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Cordoba. Instituto de Investigaciones Agrobiotecnologicas.; ArgentinaFil: Lopez, Gaston Alberto. Universidad Nacional de Rio Cuarto. Facultad de Cs.exactas Fisicoquimicas y Naturales. Instituto de Investigaciones Agrobiotecnologicas. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Cordoba. Instituto de Investigaciones Agrobiotecnologicas.; ArgentinaFil: Nievas, Sofia Mariela. Universidad Nacional de Rio Cuarto. Facultad de Cs.exactas Fisicoquimicas y Naturales. Instituto de Investigaciones Agrobiotecnologicas. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Cordoba. Instituto de Investigaciones Agrobiotecnologicas.; ArgentinaFil: Coniglio, Nayla Anahí. Universidad Nacional de Rio Cuarto. Facultad de Cs.exactas Fisicoquimicas y Naturales. Instituto de Investigaciones Agrobiotecnologicas. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Cordoba. Instituto de Investigaciones Agrobiotecnologicas.; ArgentinaFil: Halliday, N.. Science and Technology Facilities Council of Nottingham. Rutherford Appleton Laboratory; Reino UnidoFil: Cámara, M.. Science and Technology Facilities Council of Nottingham. Rutherford Appleton Laboratory; Reino UnidoFil: Cassán, F.. Universidad Nacional de Rio Cuarto. Facultad de Cs.exactas Fisicoquimicas y Naturales. Instituto de Investigaciones Agrobiotecnologicas. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Cordoba. Instituto de Investigaciones Agrobiotecnologicas.; Argentin

Analysis of the denitrification pathway and greenhouse gases emissions in Bradyrhizobium sp. strains used as biofertilizers in South America

Aims: Greenhouse gases are considered as potential atmospheric pollutants, with agriculture being one of the main emission sources. The practice of inoculating soybean seeds with Bradyrhizobium sp. might contribute to nitrous oxide (NO) emissions. We analysed this capacity in five of the most used strains of Bradyrhizobium sp. in South America. Methods and Results: We analysed the denitrification pathway and NO production by Bradyrhizobium japonicum E109 and CPAC15, Bradyrhizobium diazoefficiens CPAC7 and B. elkanii SEMIA 587 and SEMIA 5019, both in free-living conditions and in symbiosis with soybean. The in silico analysis indicated the absence of nosZ genes in B. japonicum and the presence of all denitrification genes in B. diazoefficiens strains, as well as the absence of nirK, norC and nosZ genes in B. elkanii. The in planta analysis confirmed NO production under saprophytic conditions or symbiosis with soybean root nodules. In the case of symbiosis, up to 26.1 and 18.4 times higher in plants inoculated with SEMIA5019 and E109, respectively, than in those inoculated with USDA110. Conclusions: The strains E109, SEMIA 5019, CPAC15 and SEMIA 587 showed the highest NO production both as free-living cells and in symbiotic conditions in comparison with USDA110 and CPAC7, which do have the nosZ gene. Although norC and nosZ could not be identified in silico or in vitro in SEMIA 587 and SEMIA 5019, these strains showed the capacity to produce NO in our experimental conditions. Significance and Impact of the Study: This is the first report to analyse and confirm the incomplete denitrification capacity and NO production in four of the five most used strains of Bradyrhizobium sp. for soybean inoculation in South America.Financial support was obtained from the Universidad Nacional de Río Cuarto and Consejo Nacional de Investigación Científico‐Tecnológica (CONICET) from Argentina, the FEDER‐cofinanced Project AGR2012‐1968 from Consejería de Economía, Innovación y Ciencia (Junta de Andalucía, Spain) and from the CSIC‐sponsored I‐COOP2014 project COOPA20069. We thank J. Chacón for technical assistance. M.O. and J.G. are PhD students at the UNRC granted by CONICET.Peer Reviewe