Genetic Diversity and Characterization of Symbiotic Bacteria Isolated from Endemic Phaseolus Cultivars Located in Contrasting Agroecosystems in Venezuela.

Phaseolus vulgaris is a grain cultivated in vast areas of different countries. It is an excellent alternative to the other legumes in the Venezuelan diet and is of great agronomic interest due to its resistance to soil acidity, drought, and high temperatures. Phaseolus establishes symbiosis primarily with Rhizobium and Ensifer species in most countries, and this rhizobia-legume interaction has been studied in Asia, Africa, and the Americas. However, there is currently no evidence to show that rhizobia nodulate the endemic cultivars of P. vulgaris in Venezuela. Therefore, we herein investigated the phylogenetic diversity of plant growth-promoting and N2-fixing nodulating bacteria isolated from the root nodules of P. vulgaris cultivars in a different agroecosystem in Venezuela. In comparisons with other countries, higher diversity was found in isolates from P. vulgaris nodules, ranging from α- and ß-proteobacteria. Some isolates belonging to several new phylogenetic lineages within Bradyrhizobium, Ensifer, and Mesorhizobium species were also specifically isolated at some topographical regions. Additionally, some isolates exhibited tolerance to high temperature, acidity, alkaline pH, salinity stress, and high Al levels; some of these characteristics may be related to the origin of the isolates. Some isolates showed high tolerance to Al toxicity as well as strong plant growth-promoting and antifungal activities, thereby providing a promising agricultural resource for inoculating crops.

Phylogenetic Analysis of Symbiotic Bacteria Associated with Two Vigna Species under Different Agro-Ecological Conditions in Venezuela.

Vigna is a genus of legumes cultivated in specific areas of tropical countries. Species in this genus are important crops worldwide. Vigna species are of great agronomic interest in Venezuela because Vigna beans are an excellent alternative to other legumes. However, this type of crop has some cultivation issues due to sensitivity to acidic soils, high temperatures, and salinity stress, which are common in Venezuela. Vigna species establish symbioses mainly with Bradyrhizobium and Ensifer, and Vigna-rhizobia interactions have been examined in Asia, Africa, and America. However, the identities of the rhizobia associated with V. radiata and V. unguiculata in Venezuela remain unknown. In the present study, we isolated Venezuelan symbiotic rhizobia associated with Vigna species from soils with contrasting agroecosystems or from fields in Venezuela. Several types of soils were used for bacterial isolation and nodules were sampled from environments characterized by abiotic stressors, such as high temperatures, high concentrations of NaCl, and acidic or alkaline pH. Venezuelan Vigna-rhizobia were mainly fast-growing. Sequencing of several housekeeping genes showed that in contrast to other continents, Venezuelan Vigna species were nodulated by rhizobia genus including Burkholderia, containing bacteria from several new phylogenetic lineages within the genus Bradyrhizobium. Some Rhizobium and Bradyrhizobium isolates were tolerant of high salinity and Al toxicity. The stress tolerance of strains was dependent on the type of rhizobia, soil origin, and cultivation history. An isolate classified as R. phaseoli showed the highest plant biomass, nitrogen fixation, and excellent abiotic stress response, suggesting a novel promising inoculant for Vigna cultivation in Venezuela.

Burkholderia and Paraburkholderia are Predominant Soybean Rhizobial Genera in Venezuelan Soils in Different Climatic and Topographical Regions.

The climate, topography, fauna, and flora of Venezuela are highly diverse. However, limited information is currently available on the characterization of soybean rhizobia in Venezuela. To clarify the physiological and genetic diversities of soybean rhizobia in Venezuela, soybean root nodules were collected from 11 soil types located in different topographical regions. A total of 395 root nodules were collected and 120 isolates were obtained. All isolates were classified in terms of stress tolerance under different concentrations of NaCl and Al3+. The tolerance levels of isolates to NaCl and Al3+ varied. Based on sampling origins and stress tolerance levels, 44 isolates were selected for further characterization. An inoculation test indicated that all isolates showed the capacity for root nodulation on soybean. Based on multilocus sequence typing (MLST), 20 isolates were classified into the genera Rhizobium and Bradyrhizobium. The remaining 24 isolates were classified into the genus Burkholderia or Paraburkholderia. There is currently no evidence to demonstrate that the genera Burkholderia and Paraburkholderia are the predominant soybean rhizobia in agricultural fields. Of the 24 isolates classified in (Para) Burkholderia, the nodD-nodB intergenic spacer regions of 10 isolates and the nifH gene sequences of 17 isolates were closely related to the genera Rhizobium and Bradyrhizobium, respectively. The root nodulation numbers of five (Para) Burkholderia isolates were higher than those of the 20 α-rhizobia. Furthermore, among the 44 isolates tested, one Paraburkholderia isolate exhibited the highest nitrogen-fixation activity in root nodules.

Study of nitrogen fixation by tropical legumes in acid soil from venezuelan savannas using 15N

En Venezuela, alrededor del 70 por ciento de las tierras cultivables presentan limitaciones por pH ácidos y baja disponibilidad en el suelo de P y N. Estrategias sustentables para proveer N, particularmente para sistemas de bajos a medios insumos en suelos ácidos de sabanas, pudiera ser enmendado por procesos de fijación biológica de N2. El objetivo de este estudio fue estimar el potencial de fijación biológica de nitrógeno (FBN) en varias leguminosas que crecen en suelos ácidos de sabanas venezolanas a través del uso de 15N. Los resultados sugieren que Cajanus cajan es una leguminosa con alta capacidad de fijación de nitrógeno (79 por ciento de N derivado de la atmósfera), generando un balance de N positivo en el agrosistema. La estimación de FBN en todas las leguminosas estudiadas varió dependiendo del cultivo no fijador de N usado como referencia, confirmando que el uso inadecuado de un cultivo de referencia, puede resultar en estimaciones de FBN erradas, especialmente cuando el nivel de FBN es bajo. La leguminosa Indigosphera lespediciodes mostró ser una alternativa potencial para ser usada como abono verde, debido a la acumulación de N total y a la alta cantidad de N derivado de la atmósfera (62-71 por ciento). Estos resultados demuestran que en suelos ácidos es posible promover la sustentabilidad de agrosistemas a través de prácticas de manejo conservacionista

Technical considerations for the use of 15N-DNA stable-isotope probing for functional microbial activity in soils.

Stable-isotope DNA probing is a culture-independent technique that may provide a link between function and phylogeny of active microorganisms. The technique has been used in association with 13C substrates while here we evaluate feasibility and limitations of 15N-DNA stable-isotope probing (SIP) using labelled and unlabelled pure microbial cultures or soil extracts. Our results showed that (15)N-DNA probing is feasible for cultures as well as soil samples. Limitations of 15N-DNA-SIP are (a) the need for relatively large quantities of DNA to visualise bands (although molecular resolution is much higher) and (b) 15N-DNA enrichment needed to ideally be >50 at%; however, this requirement can be lowered to approx. 40 atom% 15N with pure cultures using a modified CsCl centrifugation method (140K g for 69 h). These advances in 15N-DNA-SIP methodology open new opportunities to trace active microbial populations utilising specific N substrates in situ.

Propiedades quimico-estructurales de la materia orgánica del suelo en un agro sistema de los llanos centrales venezolanos bajo diferentes prácticas de manejo / Chemical-structural properties of the organic matter of the floor in an agriculture system of the plains central Venezuelans low different handling practices

Los usos de la tierra y las prácticas agrícolas ocasionan diferencias en la estructura y composición de la materia orgánica del suelo (MOS). La abundancia relativa de compuestos orgánicos volátiles fue determinada para evaluar las propiedades químico estructurales de la MOS bajo diferentes prácticas de manejo: labranza convencional (LC), siembra directa directa (SD), siembra directa previo un pase de cincel (CSD), mantenimiento (MR) y exportación (ER) de los residuos de cosecha del suelo, maíz continuo (M-M) y una rotación interanual maíz-soya (M-S). El experimento se realizó en un suelo Tipic Hapluster Af, ubicado en la altiplanicie de los llanos centrales (9 grados 23'33''N y 66 grados 38'30''O), Venezuela. Las características químico-estructurales de MOS se determinaron a travéz de pirólisis cromatográfica de gases (PY-CG), obteniéndose fragmentos de compuestos orgánicos volátiles en suelo y extracto. Los efectos individuales de los tratamientos no generaron diferencias en la estructura molecular de MOS, que se caracterizó por promedio de compuestos humificados y microbiológicamente alterados. MRM-S y ERM-M favorecieron el enriquesimiento del suelo con material resistente a la degradación. MR presentó un menor índice de humificación y aromatización de MOS bajo estas condiciones. La disminución de furfural, ácido acético y fenol a 20 días después de la cosecha (ddc), así como de pirrolo a 35ddc, indicó una tasa alta de descomposición de MOS antes de los 20ddc. El menorus índice de mineralización ocurrió en la combinación de SDMRM-M, mientras la estructura molecular de la sustancia húmica no mostró evidencias de haber sido alterada por SDMRM-M, y LCMRM-M