SILVER NANOPARTICLES AND THE PLANT-ASSOCIATING ABILITIES OF RHIZOBIACEAE BACTERIA

The use of nanoparticle technology in consumer products has been increasing due to their broad-spectrum antimicrobial properties. Specifically, silver nanoparticles (AgNPs) can demonstrate distinct physiochemical properties compared to bulk silver, including a large surface area to volume ratio that allows for higher reactivity with bacterial cell surfaces. AgNPs are being released into the environment, including soil ecosystems through various pathways such as points of production or during disposal of silver-containing products. This raises the concern about the potential impact on beneficial soil bacteria and their surrounding ecosystems. Members of the Rhizobiaceae family play important roles in nutrient cycling and contribute to overall soil fertility and the experiments in this thesis address the potential for AgNP-mediated toxicity on these plant-associating bacteria. Respiration analysis of Bradyrhizobium japonicum, Azospirillum brasilense, and Agrobacterium tumefaciens has revealed that AgNPs can negatively impact the growth and survival of these bacterial species, with B. japonicum being the most susceptible. Additionally, swimming motility assays using B. japonicum showed a significant decrease in colony diameter when treated with AgNPs (50 ppm). A significant decrease in root colonization of Triticum aestivum roots by A. brasilense was observed as AgNP treatment concentrations increased. Although some of the experiments could not be completed, taken together, these experiments and the research reported herein highlights the potential toxicological effects of AgNPs on bacterial species vital to the growth and health of agriculturally important crops.

SILVER NANOPARTICLES AND THE PLANT-ASSOCIATING ABILITIES OF RHIZOBIACEAE BACTERIA

The use of nanoparticle technology in consumer products has been increasing due to their broad-spectrum antimicrobial properties. Specifically, silver nanoparticles (AgNPs) can demonstrate distinct physiochemical properties compared to bulk silver, including a large surface area to volume ratio that allows for higher reactivity with bacterial cell surfaces. AgNPs are being released into the environment, including soil ecosystems through various pathways such as points of production or during disposal of silver-containing products. This raises the concern about the potential impact on beneficial soil bacteria and their surrounding ecosystems. Members of the Rhizobiaceae family play important roles in nutrient cycling and contribute to overall soil fertility and the experiments in this thesis address the potential for AgNP-mediated toxicity on these plant-associating bacteria. Respiration analysis of Bradyrhizobium japonicum, Azospirillum brasilense, and Agrobacterium tumefaciens has revealed that AgNPs can negatively impact the growth and survival of these bacterial species, with B. japonicum being the most susceptible. Additionally, swimming motility assays using B. japonicum showed a significant decrease in colony diameter when treated with AgNPs (50 ppm). A significant decrease in root colonization of Triticum aestivum roots by A. brasilense was observed as AgNP treatment concentrations increased. Although some of the experiments could not be completed, taken together, these experiments and the research reported herein highlights the potential toxicological effects of AgNPs on bacterial species vital to the growth and health of agriculturally important crops.

Classificacao taxonomica, baseada na caracterizacao molecular, das estirpes de rizobio recomendadas para a culturas da soja e do feijoeiro.

As culturas da soja [Glycine max (L.) Merril] e do feijoeiro (Phaseolus vulgaris) sao de grande importancia economica e social para o Brasil e ambas podem ser capazes de suprir suas necessidades do nutriente nitrogenio pela simbiose com bacterias da familia Rhizobiaceae. Para garantir a maximizacao do processo biologico, porem, deve-se proceder a inoculacao das sementes com estirpes de rizobio eficientes e competitivas, recomendadas pela pesquisa. No Brasil, sao comercializadas, anualmente, cerca de 13 milhoes de doses de inoculantes, sendo 99% destinadas a cultura da soja. Neste trabalho, determinou-se a posicao taxonomica das estirpes de rizobio recomendadas comercialmente para as duas culturas por duas tecnicas, o RFLP-PCR da regiao correspondente ao 16S rRNA (regiao conservada entre bacterias mas suficientemente variavel e carregando informacoes que permitem a determinacao das relacoes filogeneticas entre bacterias) e o sequenciamento parcial dos genes desta regiao. O sequenciamento parcial permitiu definir que duas das estirpes recomendadas para a cultura da soja, SEMIA 587 e SEMIA 5019, pertencem a especie Bradyrhizobium elkanii e as duas outras, SEMIA 5079 e SEMIA 5080, a especie B. japonicum. As duas estirpes recomendadas para a cultura do feijoeiro, SEMIA 4077 e SEMIA 4080, pertencem a especie Rhizobium tropici. As sequencias obtidas para essas estirpes foram depositadas no banco mundial de genes.

Evolución de poblaciones de Rhizobium Meliloti en turba

p.139-144

Selección de cepas de Rhizobium japonicum para la inoculación de soja en el Noroeste Argentino

p.255-261

Evaluation of the biotechnological potential of Rhizobium tropici strains for exopolysaccharide production

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Fijación biológica de nitrógeno por cuatro fabáceas en suelos ácidos de Tabasco, México

La fijación biológica de nitrógeno (FBN) es un proceso que ocurre en la naturaleza y representa la fuente de N más barata para los suelos ácidos del trópico húmedo. El objetivo de este trabajo fue cuantificar la cantidad de N fijado por cuatro especies de fabáceas a través de técnicas isotópicas con 15N, en un suelo de sabana en México. Los tratamientos se establecieron bajo un diseño de bloques completos al azar, con cuatro repeticiones. Las variables evaluadas fueron; biomasa fresca (BF), materia seca (MS), número de nódulos (NN), masa seca de nódulos (MSN), Nitrógeno total (Nt) y Nitrógeno fijado biológicamente (Nf). Los resultados muestran que Mucuna deerengiana L. presentó mayor producción de BF y MS (17,50 y 5,47 Mg ha-1) así como MSN (58,79 mg planta-1) y también mayor contenido de Nt y Nf (526,94 y 522,11 kg ha-1) respectivamente, en comparación con Cajanus cajan L., Phaseolus lunatus L. y Sesbania emerus L., especies que mostraron valores bajos en dichas variables. Se concluye que Mucuna deerengiana L. tolera bien los factores desfavorables que predominan en los suelos ácidos y por ello expresa una eficiencia superior a 500 kg ha-1 de Nf; se considera adecuada para aumentar el nivel de nitrógeno en los suelos de sabana sin aplicar fertilizantes químicos.

Functional and expression analysis of the metal-inducible dmeRF system from Rhizobium legumionosarum bv. viciae

A gene encoding a homolog to the cation diffusion facilitator protein DmeF from Cupriavidus metallidurans has been identified in the genome of Rhizobium leguminosarum UPM791. The R. leguminosarum dmeF gene is located downstream of an open reading frame (designated dmeR) encoding a protein homologous to the nickel- and cobalt-responsive transcriptional regulator RcnR from Escherichia coli. Analysis of gene expression showed that the R. leguminosarum dmeRF genes are organized as a transcriptional unit whose expression is strongly induced by nickel and cobalt ions, likely by alleviating the repressor activity of DmeR on dmeRF transcription. An R. leguminosarum dmeRF mutant strain displayed increased sensitivity to Co(II) and Ni(II), whereas no alterations of its resistance to Cd(II), Cu(II), or Zn(II) were observed. A decrease of symbiotic performance was observed when pea plants inoculated with an R. leguminosarum dmeRF deletion mutant strain were grown in the presence of high concentrations of nickel and cobalt. The same mutant induced significantly lower activity levels of NiFe hydrogenase in microaerobic cultures. These results indicate that the R. leguminosarum DmeRF system is a metal-responsive efflux mechanism acting as a key element for metal homeostasis in R. leguminosarum under free-living and symbiotic conditions. The presence of similar dmeRF gene clusters in other Rhizobiaceae suggests that the dmeRF system is a conserved mechanism for metal tolerance in legume endosymbiotic bacteria.