RESUMEN
Recently, the Rhizobium-legume symbiotic interaction has been proposed as an interesting tool in bioremediation. However, little is known about the effect of most common contaminants on this process. The phytotoxic effects of arsenic on nodulation of Medicago sativa have been examined in vitro using the highly arsenic resistant and symbiotically effective Sinorhizobium sp. strain MA11. The bacteria were able to grow on plates containing As concentrations as high as 10 mM. Nevertheless, as little as 25-35 microM arsenite produced a 75% decrease in the total number of nodules, due to a 90% reduction in the number of rhizobial infections, as could be determined using the strain MA11 carrying a lacZ reporter gene. This effect was associated to root hair damage and a shorter infective root zone. However, once nodulation was established nodule development seemed to continue normally, although earlier senescence could be observed in nodules of arsenic-grown plants.
Asunto(s)
Arsénico/toxicidad , Carcinógenos Ambientales/toxicidad , Medicago sativa/efectos de los fármacos , Sinorhizobium/efectos de los fármacos , Contaminantes del Suelo/toxicidad , Biodegradación Ambiental , Ecología/métodos , Medicago sativa/crecimiento & desarrollo , Medicago sativa/microbiología , Fijación del Nitrógeno , Nódulos de las Raíces de las Plantas/efectos de los fármacos , Nódulos de las Raíces de las Plantas/crecimiento & desarrollo , Sinorhizobium/metabolismo , Simbiosis/efectos de los fármacosRESUMEN
In Morocco, storage reservoirs are particular systems of water supply in rural areas. These reservoirs are fed with rainwater and/or directly from the river, which are very contaminated by several pathogenic bacteria. They are used without any treatment as a drinking water by the surrounding population. In this context, the aim of this study is to evaluate the impact of consuming contaminated water stored in reservoirs on health status for six rural communities located in Assif El Mal, Southern East of Marrakech. This was investigated using a classical methodology based on population survey and by molecular approach using PCR-DGGE technique to determine the intestinal bacterial diversity of consumers. The survey showed that, the residents of the studied area suffered from numerous health problems (diarrheal diseases, vomiting or hepatitis A) due to the lack of waste management infrastructures. The consumer's stool analysis by molecular approach revealed that numbers of Escherichia coli, Aeromonas hydrophila and Clostridia, were significantly higher in the diarrheal feces. In addition, PCR-DGGE study of the prevalence and distribution of bacteria causing human diseases, confirmed that, there is a relationship between water bacterial contaminations of storage reservoirs and microbial disease related health status. Therefore, water reservoir consumption is assumed to be the mean way of exposure for this population. It's clear that this approach gives a very helpful tool to confirm without any doubt the relationship between water bacterial contamination and health status.
RESUMEN
The presence of cyanotoxins, mainly microcystins (MCs), in surface freshwater represents a serious health risk to aquatic organisms living in the water body, as well as terrestrial animals and plants that are in contact with contaminated water. Consequently, the use of MCs contaminated water for irrigation represents a hazard for cultivated plants and could induce severe economical losses due to crops' yield reduction. The experimental approach undertaken in this work was exposing Vicia faba seedlings (inoculated with a Rhizobium strain resistant to MCs), to water supplemented with cyanobacterial crude extract containing total microcystins at a concentration of 50 and 100 µg/L (environmental relevant concentrations of MCs dissolved in the raw irrigation water from Lalla Takerkoust Lake-Marrakesh region). After chronic MCs exposure (2 months), biological and physiological parameters (plant growth, nitrogen uptake, mineral assimilation, and oxidative defense mechanisms) were evaluated. The results obtained showed evidence that chronic exposure to cyanobacterial bloom extract containing MCs strongly affected the physiological and biological plants activities; reduction of dry matter, photosynthetic activity, nodule number, and nitrogen assimilation. At the same time, an increase of oxidative stress was observed, as deduced from a significant increase of the activities of peroxidase, catalase, polyphenoloxidase, and phenylalanine ammonia lyase in leaves, roots, and nodules of faba bean plants exposed to cyanotoxins, especially at 100 µg/L of MCs. This experimentation constitutes a simulation of the situation related to cyanotoxins chronic exposure of seedlings-plants via the contaminated irrigation water. For this reason, once should take into consideration the possibility of contamination of agricultural crops and the quality of irrigation water should be by the way monitored for cyanotoxins biohazard.
Asunto(s)
Mezclas Complejas/farmacología , Cianobacterias , Microcistinas/farmacología , Rhizobium/efectos de los fármacos , Simbiosis/efectos de los fármacos , Vicia/efectos de los fármacos , Catalasa/metabolismo , Catecol Oxidasa/metabolismo , Peroxidasa/metabolismo , Fenoles/metabolismo , Fenilanina Amoníaco-Liasa/metabolismo , Fotosíntesis/efectos de los fármacos , Hojas de la Planta/efectos de los fármacos , Hojas de la Planta/metabolismo , Raíces de Plantas/efectos de los fármacos , Raíces de Plantas/metabolismo , Rhizobium/fisiología , Vicia/fisiologíaRESUMEN
BACKGROUND: Rhizobium-Legume symbiosis is an attractive biological process that has been studied for decades because of its importance in agriculture. However, this system has undergone extensive study and although many of the major factors underpinning the process have been discovered using traditional methods, much remains to be discovered. RESULTS: Here we present an analysis of the 'Symbiosis Interactome' using novel computational methods in order to address the complex dynamic interactions between proteins involved in the symbiosis of the model bacteria Sinorhizobium meliloti with its plant hosts. Our study constitutes the first large-scale analysis attempting to reconstruct this complex biological process, and to identify novel proteins involved in establishing symbiosis. We identified 263 novel proteins potentially associated with the Symbiosis Interactome. The topology of the Symbiosis Interactome was used to guide experimental techniques attempting to validate novel proteins involved in different stages of symbiosis. The contribution of a set of novel proteins was tested analyzing the symbiotic properties of several S. meliloti mutants. We found mutants with altered symbiotic phenotypes suggesting novel proteins that provide key complementary roles for symbiosis. CONCLUSION: Our 'systems-based model' represents a novel framework for studying host-microbe interactions, provides a theoretical basis for further experimental validations, and can also be applied to the study of other complex processes such as diseases.
Asunto(s)
Biología Computacional , Sinorhizobium meliloti/metabolismo , Simbiosis , Proteínas Bacterianas/metabolismo , Bases de Datos de Proteínas , Evolución Molecular , Genoma Bacteriano , Medicago sativa/metabolismo , Medicago sativa/fisiología , Modelos Biológicos , Sinorhizobium meliloti/genética , Sinorhizobium meliloti/fisiologíaRESUMEN
Tip growth is a specialized type of polar growth where new cell wall is deposited in a localized region of the cell, the growing tip. These cells show a characteristic zonation, with a high accumulation of secretory vesicles containing cell wall components at the tip, followed by an organelle-enriched zone. MsPG3 is a Medicago sativa polygalacturonase gene isolated in our laboratory, specifically expressed during the interaction of this plant with its symbiotic partner Sinorhizobium meliloti and which might participate in tip growth processes during symbiosis. We have used MsPG3-GFP fusions to study in vivo protein transport processes and localization during root hair growth. Different MsPG3-GFP fusions were expressed in Medicago truncatula'hairy roots' following a protocol developed for this study and also tested by transient expression in onion epidermal cells. Preferential accumulation of an MsPG3-GFP fusion protein in the tip of the growing root hair at different developmental stages was found, confirming the delivery of MsPG3 to the newly synthesized cell wall. This indicates that this protein may participate in tip growth processes during symbiosis and, in addition, that this fusion could be a useful tool to study this process in plants.
Asunto(s)
Proteínas Luminiscentes/genética , Medicago/genética , Poligalacturonasa/genética , Técnicas de Transferencia de Gen , Genes Reporteros , Proteínas Fluorescentes Verdes , Medicago/metabolismo , Cebollas/metabolismo , Epidermis de la Planta/metabolismo , Raíces de Plantas/metabolismo , Transporte de Proteínas/fisiología , Proteínas Recombinantes de Fusión/genéticaRESUMEN
While the biology of nitrogen-fixing root nodules has been extensively studied, little is known about the evolutionary events that predisposed legume plants to form symbiosis with rhizobia. We have studied the presence and the expression of two pectic gene families in Medicago, polygalacturonases (PGs) and pectin methyl esterases (PMEs) during the early steps of the Sinorhizobium meliloti-Medicago interaction and compared them with related pollen-specific genes. First, we have compared the expression of MsPG3, a PG gene specifically expressed during the symbiotic interaction, with the expression of MsPG11, a highly homologous pollen-specific gene, using promoter-gus fusions in transgenic M. truncatula and tobacco plants. These results demonstrated that the symbiotic promoter functions as a pollen-specific promoter in the non-legume host. Second, we have identified the presence of a gene family of at least eight differentially expressed PMEs in Medicago. One subfamily is represented by one symbiotic gene (MtPER) and two pollen-expressed genes (MtPEF1 and MtPEF2) that are clustered in the M. truncatula genome. The promoter-gus studies presented in this work and the homology between plant PGs, together with the analysis of the PME locus structure and MtPER expression studies, suggest that the symbiotic MsPG3 and MtPER could have as ancestors pollen-expressed genes involved in polar tip growth processes during pollen tube elongation. Moreover, they could have been recruited after gene duplication in the symbiotic interaction to facilitate polar tip growth during infection thread formation.