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Bacterial Community of Rhizosphere Associated to the Annual Halophyte

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Bacterial Community of Rhizosphere Associated to the Annual Halophyte BACTERIAL COMMUNITY OF RHIZOSPHERE ASSOCIATED TO THE ANNUAL HALOPHYTE Salicornia bigelovii (Torr.) Comunidad Bacteriana de la Rizosfera Asociada a la Halófita Anual Salicornia bigelovii (Torr.) Edgar O. Rueda-Puente1‡, Thelma Castellanos-Cervantes2, José Luis Díaz de León-Álvarez3, Pablo Preciado-Rangel4 and Gustavo Almaguer-Vargas5 SUMMARY with S. bigelovii, a novel halophyte crop. Further studies on diversity of the microbial community in the Salicornia The specie Salicornia bigelovii (Chenopodiaceae) bigelovii rhizosphere are required to clarify and describe is a halophyte with promising biological and commercial this ecological association under saline conditions. value as a natural resource from arid zones. However, its productivity depends on the nitrogen supply. Nitrogen Index words: plant growth-promoting bacteria fixation by bacteria associated with the roots of S. (PGPB), biofertilizer, soil ecology. bigelovii is a determinant source of available nitrogen. However, the diversity in Salicornia’s rhizosphere is RESUMEN unknown. In order to increase the knowledge of the bacterial diversity associated with the S. bigelovii La especie Salicornia bigelovii (Chenopodiaceae) rhizosphere, polymerase chain reaction (PCR) and single- es una halófita con un promisorio valor biológico y strand conformation polymorphism (SSCP) analysis was comercial en zonas áridas. Sin embargo, su productividad performed. This study was conducted in La Paz, BCS, depende de la aportación de nitrógeno. La fijación de Mexico. A random sample of 25 S. bigelovii plants at nitrógeno por bacterias asociadas a la raíz de S. bigelovii, the flowering stage was collected from 6 areas along es una fuente determinante para disponer de nitrógeno. the coastal line of La Paz Bay in the southern portion of Sin embargo, la diversidad de estos microorganismos en the Baja California Peninsula. DNA extractions of Salicornia se desconoce. Por lo anterior, el objetivo del bacterial DNA through amplification (PCR) of the 16S presente trabajo consistió en conocer la comunidad rDNA, SSCP-profiles analysis, isolation of selected bacteriana, asociada a la rizósfera de S. bigelovii, bands of SSCP-profiles, cloning, and sequences were mediante la técnica de reacción en cadena de la carried out. The SSCP technique reflected the bacterial polimerasa (PCR) y la técnica de conformación composition evidencing the available diversity and also polimórfica de cadena simple (SSCP). El estudio fue the dominant populations. The results obtained reflect desarrollado en Bahía de La Paz, BCS México, mediante the presence of Rhizobium spp. and Bacillus spp. This un muestreo al azar de 25 plantas de S. bigelovii en is the first report of these microorganisms associated etapa fenológica de floración, las cuales fueron colectadas de 6 áreas del sistema costero. Se realizaron 1 Universidad de Sonora, Campus Santa Ana. Carretera Internacional extracciones de ADN de la población bacteriana a través y Avenida 16 de Septiembre s/n, Col. La Loma. 84600 Ciudad Santa de amplificación por PCR del 16 S rDNA, análisis Ana, Sonora, México. mediante SSCP, aislamiento de bandas seleccionadas ‡ Autor responsable ([email protected]) de SSCP, clonación y, secuenciación. Los resultados 2 Centro de Investigaciones Biológicas del Noroeste. Mar Bermejo No. 195, Col. Playa Palo Santa Rita. Apartado Postal 128. 23090. mostraron que la técnica SSCP evidencia una diversa La Paz, Baja California Sur, México. composición bacteriana con poblaciones dominantes de 3 Universidad Autónoma de Baja California Sur. Apartado Postal las cuales los resultados obtenidos indican la presencia 128. 23080 La Paz, Baja California Sur, México. de Rhizobium spp. y Bacillus spp. Este es el primer 4 Instituto Tecnológico de Torreón. km 7.5 carretera Torreón-San informe de este tipo de microorganismos asociados a S. Pedro. 27170 ejido Ana, Torreón, Coahuila, México. 5 Universidad Autónoma Chapingo. 56230 Chapingo, estado de bigelovii. Estudios relacionados con la diversidad México. microbiana en rizósfera de Salicornia bigelovii deben ser desarrollados para clarificar y describir las Recibido: abril del 2010. Aceptado: noviembre de 2010. asociaciones ecológicas presentes bajo condiciones de Publicado como nota de investigación en Terra Latinoamericana 28: 345-353. salinidad. 345 346 TERRA LATINOAMERICANA VOLUMEN 28 NÚMERO 4, 2010 Palabras clave: bacterias promotoras del crecimiento populations of the same species. For multiple vegetal (BPCV), biofertilizantes, ecología del suelo. environmental and economic purposes, it is important to increase the populations of salt-tolerant, nitrogen-fixing INTRODUCTION bacteria (Zahran, 1999), so that potential novel crops, such as Salicornia bigelovii, which has already been Halophytes, particularly Salicornia bigelovii considered a new plant resource in arid region agriculture (Chenopodiaceae), are promising plant resources in arid (Bashan et al., 1992), can thrive. coastal zones because they are able to tolerate high saline Few studies on soil ecology and bacterial diversity conditions. Such salt-adapted plant resources are within the rhizosphere of salt marsh plants are known commonly found along coastal plains (Glenn et al., 1991). and available. In previous studies, one nitrogen fixing These important plants hold the potential to be bacterium was isolated from Salicornia bigelovii incorporated into traditional agriculture to help support rhizosphere through conventional techniques, using an the agricultural economy in coastal areas (Glenn et al., N-free media: OAB (Reinhold et al., 1987) and Rennie’s 1995). In the states of Baja California Sur and Sonora, medium (Rennie, 1981), evidencing Klebsiella Mexico, S. bigelovii is widely distributed along the pneumoniae as a potential plant-growth-promoting- coasts. It has been suggested elsewhere that this rhizobacterium (Rueda et al., 2003; 2004). Other halophyte has real prospects for commercial exploitation reported studies were limited to the study of mycoflora, (Glenn et al., 1994; Troyo et al., 1994). However, in the mainly S. europaea (Ito et al., 1999). southern part of these states, halophyte productivity is This work focused on the composition of the natural limited by a lack of available nitrogen. This condition bacterial diversity inhabiting the rhizosphere of the affects growth, reproduction, and nutrient nitrogen levels halophyte Salicornia bigelovii, by means of the single- within the plant biomass (Rueda et al., 2004). strand conformation polymorphism (SSCP) technique. Traditionally, to solve soil nitrogen deficiency, farmers apply synthetic fertilizers, such as urea, ammonium MATERIALS AND METHODS sulfate, and others. However, their indiscriminate use severely damages soil microbiological flora (Banwari and Microbial DNA Isolation Rao, 1990; Al-Nahidh and Gomah, 1991; Akhavan et al., 1991), and also gradually increases the level of soil salinity. Plant and soil samples of 26 plants were collected Another option to improve the availability of nitrogen at flowering stage from six areas along the southern is the use of plant-growth-promoting-bacteria (PGPB), shore of La Paz Bay, in Baja California Sur, northwest which are gaining acceptance worldwide; a large number Mexico, 17 km west of La Paz City (sample 1: Latitude of bacterial strains have been isolated and evaluated 24° 8’ 16.36” N, Longitude 110° 25’ 33.92” O; sample 2: primarily for improving plant growth (Kloeper et al., Latitude 24° 8’ 14.05” N, Longitude 110° 25’ 32.19” O; 1991). Strains that fail to demonstrate a positive effect sample 3: Latitude 24° 7’ 55.56” N, Longitude 110° 25’ or that induce a negative plant response are routinely 8.10” O; sample 4: Latitude 24° 7’ 50.44” N, Longitude discarded and their negative effects are seldom reported 110° 25’ 12.44” O; sample 5: Latitude 24° 7’ 48.88” N, (Nehl et al., 1997). Longitude 110° 25’ 6.31” O; sample 6: Latitude 24° 7’ These strains can be beneficial or harmful (Nehl 30.17” N, Longitude 110° 25’ 15.36” O). et al., 1997), saprophytic becoming pathogenic by Conventional soil and plant physical-chemical changing the host, or any of these under different analysis were conducted to determine texture class (using microbe-host relationships (Pimentel et al., 1991). Some Bouyoucos densimeter), organic matter concentration strains may exhibit beneficial effects on one plant species (by titration with ferrous sulfate) salinity of saturated but may exert harmful or variable effects on others paste (using the Weatston electrical conductivity bridge), (O’neil et al., 1992). Because of the variability in strain’s sodium balance expressed as SAR (numerical features, some of the groups or strains do not hold calculation of the sodium adsorption relationship properties that enhance growth in some plant species. parameter, quantifying Ca2+ and Mg2+ by EDTA method), In this sense, it becomes relevant to know what kind of and the concentration of major nutrients through bacterial population is present in the plant rhizosphere bromatological analysis. The general characteristics to isolate them for inoculation in other individuals or of the soil present in the six areas sampled are cited in RUEDA ET AL. BACTERIAL COMMUNITY DIVERSITY OF RHIZOSPHERE 347 Table 1. La Paz Bay has different geographical and the phosphorilated strand was performed by means of climatic conditions. The Bay area and the coast are the enzyme exonuclease (LAMBDA) in order to obtain desert, warm (19 ± 4 °C) in the summer and cool in simple strands. Electrophoresis
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