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      Bacterial community of rhizosphere associated to the annual halophyte Salicornia bigelovii (Torr.) Translated title: Comunidad bacteriana de la rizosfera asociada a la halófita anual Salicornia bigelovii (Torr.)

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          Abstract

          The specie Salicornia bigelovii (Chenopodiaceae) is a halophyte with promising biological and commercial value as a natural resource from arid zones. However, its productivity depends on the nitrogen supply. Nitrogen fixation by bacteria associated with the roots of S. bigelovii is a determinant source of available nitrogen. However, the diversity in Salicomia's rhizosphere is unknown. In order to increase the knowledge of the bacterial diversity associated with the S. bigelovii rhizosphere, polymerase chain reaction (PCR) and single-strand conformation polymorphism (SSCP) analysis was performed. This study was conducted in La Paz, BCS, Mexico. A random sample of 25 S. bigelovii plants at the flowering stage was collected from 6 areas along the coastal line of La Paz Bay in the southern portion of the Baja California Peninsula. DNA extractions of bacterial DNA through amplification (PCR) of the 16S rDNA, SSCP-profiles analysis, isolation of selected bands of SSCP-profiles, cloning, and sequences were carried out. The SSCP technique reflected the bacterial composition evidencing the available diversity and also the dominant populations. The results obtained reflect the presence of Rhizobium spp. and Bacillus spp. This is the first report of these microorganisms associated with S. bigelovii, a novel halophyte crop. Further studies on diversity of the microbial community in the Salicornia bigelovii rhizosphere are required to clarify and describe this ecological association under saline conditions.

          Translated abstract

          La especie Salicornia bigelovii (Chenopodiaceae) es una halófita con un promisorio valor biológico y comercial en zonas áridas. Sin embargo, su productividad depende de la aportación de nitrógeno. La fijación de nitrógeno por bacterias asociadas a la raíz de S. bigelovii, es una fuente determinante para disponer de nitrógeno. Sin embargo, la diversidad de estos microorganismos en Salicornia se desconoce. Por lo anterior, el objetivo del presente trabajo consistió en conocer la comunidad bacteriana, asociada a la rizósfera de S. bigelovii, mediante la técnica de reacción en cadena de la polimerasa (PCR) y la técnica de conformación polimórfica de cadena simple (SSCP). El estudio fue desarrollado en Bahía de La Paz, BCS México, mediante un muestreo al azar de 25 plantas de S. bigelovii en etapa fenológica de floración, las cuales fueron colectadas de 6 áreas del sistema costero. Se realizaron extracciones de ADN de la población bacteriana a través de amplificación por PCR del 16 S rDNA, análisis mediante SSCP, aislamiento de bandas seleccionadas de SSCP, clonación y, secuenciación. Los resultados mostraron que la técnica SSCP evidencia una diversa composición bacteriana con poblaciones dominantes de las cuales los resultados obtenidos indican la presencia de Rhizobium spp. y Bacillus spp. Este es el primer informe de este tipo de microorganismos asociados a S. bigelovii. Estudios relacionados con la diversidad microbiana en rizósfera de Salicornia bigelovii deben ser desarrollados para clarificar y describir las asociaciones ecológicas presentes bajo condiciones de salinidad.

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          Fast and sensitive silver staining of DNA in polyacrylamide gels

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            Endophytic colonization and in planta nitrogen fixation by a Herbaspirillum sp. isolated from wild rice species.

            Nitrogen-fixing bacteria were isolated from the stems of wild and cultivated rice on a modified Rennie medium. Based on 16S ribosomal DNA (rDNA) sequences, the diazotrophic isolates were phylogenetically close to four genera: Herbaspirillum, Ideonella, Enterobacter, and Azospirillum. Phenotypic properties and signature sequences of 16S rDNA indicated that three isolates (B65, B501, and B512) belong to the Herbaspirillum genus. To examine whether Herbaspirillum sp. strain B501 isolated from wild rice, Oryza officinalis, endophytically colonizes rice plants, the gfp gene encoding green fluorescent protein (GFP) was introduced into the bacteria. Observations by fluorescence stereomicroscopy showed that the GFP-tagged bacteria colonized shoots and seeds of aseptically grown seedlings of the original wild rice after inoculation of the seeds. Conversely, for cultivated rice Oryza sativa, no GFP fluorescence was observed for shoots and only weak signals were observed for seeds. Observations by fluorescence and electron microscopy revealed that Herbaspirillum sp. strain B501 colonized mainly intercellular spaces in the leaves of wild rice. Colony counts of surface-sterilized rice seedlings inoculated with the GFP-tagged bacteria indicated significantly more bacterial populations inside the original wild rice than in cultivated rice varieties. Moreover, after bacterial inoculation, in planta nitrogen fixation in young seedlings of wild rice, O. officinalis, was detected by the acetylene reduction and (15)N(2) gas incorporation assays. Therefore, we conclude that Herbaspirillum sp. strain B501 is a diazotrophic endophyte compatible with wild rice, particularly O. officinalis.
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              Classification of rhizobia based on nodC and nifH gene analysis reveals a close phylogenetic relationship among Phaseolus vulgaris symbionts.

              The nodC and nifH genes were characterized in a collection of 83 rhizobial strains which represented 23 recognized species distributed in the genera Rhizobium, Sinorhizobium, Mesorhizobium and Bradyrhizobium, as well as unclassified rhizobia from various host legumes. Conserved primers were designed from available nucleotide sequences and were able to amplify nodC and nifH fragments of about 930 bp and 780 bp, respectively, from most of the strains investigated. RFLP analysis of the PCR products resulted in a classification of these rhizobia which was in general well-correlated with their known host range and independent of their taxonomic status. The nodC and nifH fragments were sequenced for representative strains belonging to different genera and species, most of which originated from Phaselous vulgaris nodules. Phylogenetic trees were constructed and revealed close relationships among symbiotic genes of the Phaseolus symbionts, irrespective of their 16S-rDNA-based classification. The nodC and nifH phylogenies were generally similar, but cases of incongruence were detected, suggesting that genetic rearrangements have occurred in the course of evolution. The results support the view that lateral genetic transfer across rhizobial species and, in some instances, across Rhizobium and Sinorhizobium genera plays a role in diversification and in structuring the natural populations of rhizobia.
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                Author and article information

                Journal
                tl
                Terra Latinoamericana
                Terra Latinoam
                Sociedad Mexicana de la Ciencia del Suelo A.C. (Chapingo, Estado de México, Mexico )
                0187-5779
                2395-8030
                December 2010
                : 28
                : 4
                : 345-353
                Affiliations
                [01] Santa Ana Sonora orgnameUniversidad de Sonora orgdiv1Campus Santa Ana México erueda04@ 123456santana.uson.mx
                [03] La Paz Baja California Sur orgnameUniversidad Autónoma de Baja California Sur México
                [05] Chapingo Estado de México orgnameUniversidad Autónoma Chapingo México
                [04] Torreón Coahuila orgnameInstituto Tecnológico de Torreón México
                [02] La Paz Baja California Sur orgnameCentro de Investigaciones Biológicas del Noroeste S.C. México
                Article
                S0187-57792010000400006 S0187-5779(10)02800400006
                8ecfb606-b98e-4cdb-83c9-347998baffd7

                This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.

                History
                : November 2010
                : April 2010
                Page count
                Figures: 0, Tables: 0, Equations: 0, References: 50, Pages: 9
                Product

                SciELO Mexico

                Categories
                Division II

                ecología del suelo,biofertilizantes,bacterias promotoras del crecimiento vegetal (BPCV),soil ecology,biofertilizer,plant growth-promoting bacteria (PGPB)

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