Effects of nutritional and environmental conditions on Sinorhizobium meliloti biofilm formation

Rhizobia are non-spore-forming soil bacteria that fix atmospheric nitrogen into ammonia in a symbiosis with legume roots. However, in the absence of a legume host, rhizobia manage to survive and hence must have evolved strategies to adapt to diverse environmental conditions. The capacity to respond to variations in nutrient availability enables the persistence of rhizobial species in soil, and consequently improves their ability to colonize and to survive in the host plant. Rhizobia, like many other soil bacteria, persist in nature most likely in sessile communities known as biofilms, which are most often composed of multiple microbial species. We have been employing in vitro assays to study environmental parameters that might influence biofilm formation in the Medicago symbiont Sinorhizobium meliloti. These parameters include carbon source, amount of nitrate, phosphate, calcium and magnesium as well as the effects of osmolarity and pH. The microtiter plate assay facilitates the detection of subtle differences in rhizobial biofilms in response to these parameters, thereby providing insight into how environmental stress or nutritional status influences rhizobial survival. Nutrients such as sucrose, phosphate and calcium enhance biofilm formation as their concentrations increase, whereas extreme temperatures and pH negatively affect biofilm formation.Fil: Rinaudi, Luciana Veronica. Universidad Nacional de Río Cuarto; ArgentinaFil: Fujishige, Nancy A.. University of California; Estados UnidosFil: Hirsch, Ann M.. University of California; Estados UnidosFil: Banchio, Erika. Universidad Nacional de Río Cuarto; ArgentinaFil: Zorreguieta, Ángeles. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Giordano, Walter Fabian. Universidad Nacional de Río Cuarto; Argentin

An integrated view of biofilm formation in Rhizobia

Biofilms are bacterial communities enclosed within an extracellular matrix of polysaccharides produced by the bacteria, which adhere to a living or an inert macrosurface. In nature, biofilms constitute a protected growth modality allowing bacteria to survive in hostile environments. Studies of environmental isolates have revealed a highly ordered, three-dimensional organization of the extracellular matrix, which has important implications for biofilm physiology. The zone of soil immediately surrounding a plant root where complex biological and ecological processes occur, termed rhizosphere, forms an environment that fulfills the requirements for biofilm formation, including sufficient moisture and supply of nutrients, which are provided by the plant. Biofilm formation on plants appears to be associated with symbiotic and pathogenic responses, but it is unclear how plants regulate the association. Biofilms function as structures resistant against stress factors such as desiccation, UV radiation, predation, and antibiosis, which help create protective niches for rhizobia. However, the role of biofilms in rhizobial–legume symbiosis remains to be clarified. Here, the mechanisms involved in bacterial biofilm formation and attachment on plant roots, and the relation of these mechanisms to rhizobial function and survival are reviewed.Fil: Rinaudi, Luciana Veronica. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Biología Molecular; ArgentinaFil: Giordano, Walter Fabian. Universidad Nacional de Rio Cuarto. Facultad de Cs.exactas Fisicoquimicas y Naturales. Instituto de Biotecnologia Ambiental y Salud. - Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet - Cordoba. Instituto de Biotecnologia Ambiental y Salud.; Argentin

EPS II-dependent autoaggregation of sinorhizobium meliloti planktonic cells

Planktonic cells of Sinorhizobium meliloti, a Gram-negative symbiotic bacterium, display autoaggregation under static conditions. ExpR is a LuxR-type regulator that controls many functions in S. meliloti, including synthesis of two exopolysaccharides, EPS I (succinoglycan) and EPS II (galactoglucan). Since exopolysaccharides are important for bacterial attachment, we studied the involvement of EPS I and II in autoaggregation of S. meliloti. Presence of an intact copy of the expR locus was shown to be necessary for autoaggregation. A mutant incapable of producing EPS I displayed autoaggregation percentage similar to that of parental strain, whereas autoaggregation was significantly lower for a mutant defective in biosynthesis of EPS II. Our findings clearly indicate that EPS II is the essential component involved in autoaggregation of planktonic S. meliloti cells, and that EPS I plays no role in such aggregation. © 2010 Springer Science+Business Media, LLC.Fil: Sorroche, Fernando Guido. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Rinaudi, Luciana Veronica. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Zorreguieta, Ángeles. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquímicas de Buenos Aires. Fundación Instituto Leloir. Instituto de Investigaciones Bioquímicas de Buenos Aires; ArgentinaFil: Giordano, Walter Fabian. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas, Fisicoquímicas y Naturales. Departamento de Biología Molecular. Sección Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentin

Analysis of mucR gene regulating biosynthesis of exopolysaccharides: implications for biofilm formation in Sinorhizobium meliloti Rm1021

Bacterial surface polysaccharides are crucial for establishment of successful rhizobia-legume symbiosis, and in most bacteria, are also critical for biofilm formation and surface colonization. In Sinorhizobium meliloti, the regulatory protein MucR controls exopolysaccharide production. To clarify the relationship between exopolysaccharide synthesis and biofilm formation, we studied mucR expression under growth conditions that influence attachment to polyvinylchloride, developed a microtiter plate assay to quantify biofilm formation in S. meliloti strain Rm1021 and mutants defective in succinoglycan (EPS I) and/or galactoglucan (EPS II) production, and analyzed expression of EPS I and EPS II genes by quantitative reverse transcriptase-PCR. Consistent with previous studies of planktonic bacteria, we found that disruption of the mucR gene in Rm1021 biofilms increased EPS II, but reduced EPS I gene expression. mucR expression was not affected by environmental conditions that influence biofilm formation on polyvinylchloride, and biofilm formation by Rm1021 was independent of exopolysaccharide synthesis. Other factors on the Rm1021 cell surface, and growth conditions, presumably regulate attachment and/or growth as a biofilm on polyvinylchloride.Fil: Rinaudi, Luciana Veronica. Universidad Nacional de Río Cuarto. Departamento de Biología Molecular; ArgentinaFil: Sorroche, Fernando Guido. Universidad Nacional de Río Cuarto. Departamento de Biología Molecular; ArgentinaFil: Zorreguieta, Angeles. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Bioquimicas de Buenos Aires; Argentina. Fundación Instituto Leloir; ArgentinaFil: Giordano, Walter Fabian. Universidad Nacional de Río Cuarto. Departamento de Biología Molecular; Argentin

Changes in composition of essential oils and volatile emissions of Minthostachys mollis, induced by leaf punctures of Liriomyza huidobrensis

Plant defensive mechanisms against herbivores include chemical changes following damage. Effects of feeding punctures produced by Liriomyza huidobrensis (pea leafminers) adult females on the plant’s dominant monoterpenes, pulegone and menthone were assessed by monitoring essential oil composition at 24, 48, and 120 h; emission of volatiles was also measured 24 and 48 h after wounding. We studied such changes in Minthostachys mollis, a Lamiaceae species native to Central Argentina with medicinal and aromatic uses. Leaf puncturing resulted in reduced menthone throughout the experiment and increased pulegone concentration in M. mollis essential oil during the first 48 h. The adjacent undamaged leaves showed similar changes, suggesting a systemic response. Composition of volatiles released from damaged leaves was also altered, most noticeably by increasing pulegone and diminishing menthone emissions. Such herbivore-induced chemical changes in aromatic plants are economically relevant, since the quality of essential oils and volatile emissions are altered.Fil: Banchio, Erika. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Valladares, Graciela Rosa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Zygadlo, Julio Alberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto Multidisciplinario de Biología Vegetal. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas Físicas y Naturales. Instituto Multidisciplinario de Biología Vegetal; ArgentinaFil: Bogino, Pablo Cesar. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Rinaudi, Luciana Veronica. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; ArgentinaFil: Giordano, Walter Fabian. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba; Argentin