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The indiscriminate use of chemical fertilizers has contributed to the deterioration of the biological, physical and chemical properties of the soil, resulting in the loss of its productive capacity. For this reason, the use of biofertilizers has emerged as a technological alternative. The objective of this research was to develop a suitable liquid fermentation system and encystment for the multiplication of Azotobacter chroococcum AC1 strain, a bacterium employed in a biofertilizer formulation produced at present by CARPOICA, Colombia. Sequential statistical designs were used to determine the conditions in the fermentation system. The interaction between agitation, aeration and pH was evaluated on the viable biomass (CFU/ml) of AC1. In addition, the encystment ability of the strain was evaluated using two encystment agents and the potential plant growth-promoting rhizobacteria (PGPR) activity was assessed by different techniques, such as nitrogen fixation by ARA, phosphate solubilization by the phospho-molybdenum-blue reaction and indolic compound production by colorimetric reaction using the Salkowski reagent. Results showed significant effects (p

Introduction: The sustainable production of pastures has become a fundamental challenge for the livestock sector where research with plant growth-promoting rhizobacteria as a viable solution, has nearly not been reported. Objective: In this study, we aimed to examine the potential to stimulate growth in Pennisetum clandestinum grass using four isolated bacterial strains from soils obtained from a Colombian tropical silvopastoral system. Methods: We previously identified genetically the strains and characterized two plant growth promoting activities. In addition, we evaluated the growth-promoting effect of the strains in Kikuyo grass under greenhouse conditions. Results: We found that the four bacterial strains were phylogenetically associated with Klebsiella sp. (strains 28P and 35P), Beijerinka sp. (37L) and Achromobacter xylosoxidans (E37), based on partial 16S rRNA gene sequencing. Moreover, the in vitro biochemical assays demonstrated that the strains exhibited some plant growth promoting mechanisms such as 1-aminocyclopropane-1-carboxylic acid deaminase activity and indole compound synthesis. Notably, bacterial inoculation under greenhouse conditions showed a positive influence on P. clandestinum growth. We found a significant (P < 0.05) effect on root and shoot length and shoot dry weight. Shoot length increased by 52 % and 30 % with 37L and 35P compared to those without inoculation treatment. Similarly, the use of 37L and 28P raised shoot dry weight values by 170 % and 131 %, respectively. In root development, inoculation with strains 37L and E37 increased root length by 134 % and 100 %, respectively. Conclusion: Beijerinckia sp. 37L was the most effective of the four strains at increasing P. clandestinum biomass and length. RESUMEN Introducción: La producción sostenible de pastos se ha convertido en un desafío fundamental para el sector ganadero, donde investigaciones con bacterias promotoras de crecimiento vegetal, como una solución viable, han sido poco reportadas. Objetivo: El objetivo de este estudio fue examinar el potencial para estimular el crecimiento del pasto Pennisetum clandestinum utilizando cuatro cepas bacterianas aisladas de suelos obtenidos de un sistema silvopastoril tropical colombiano. Métodos: Anteriormente identificamos genéticamente las cepas y caracterizamos dos actividades que promueven el crecimiento de las plantas. Además, evaluamos el efecto promotor del crecimiento de las cepas en el pasto Kikuyo en condiciones de invernadero. Resultados: Encontramos que las cuatro cepas bacterianas se asociaron filogenéticamente con Klebsiella sp. (cepas 28P y 35P), Beijerinka sp. (37L) y Achromobacter xylosoxidans (E37), basados en la secuenciación parcial del gen 16S rRNA. Además, los ensayos bioquímicos in vitro demostraron que las cepas exhibían algunos mecanismos que promueven el crecimiento de las plantas tales como la actividad de la enzima desaminasa del ácido 1-aminociclopropano-1- carboxílico, y la síntesis del compuesto indol. En particular, la inoculación bacteriana bajo condiciones de invernadero mostró una influencia positiva en el crecimiento de P. clandestinum. Encontramos un efecto significativo (P < 0.05) en la longitud de la raíz y el tallo, y el peso seco del tallo. La longitud del tallo aumentó en un 52 % y 30 % con 37L y 35P, respectivamente, en comparación con aquellos sin tratamiento de inoculación. Igualmente, el uso de las cepas 37L y 28P aumentó los valores de peso seco del tallo en un 170 y un 131 %, respectivamente. En el desarrollo de la raíz, la inoculación con las cepas 37L y E37 aumentó la longitud de la raíz en 134 y 100 %, respectivamente. Conclusión: Beijerinckia sp. 37L fue la más efectiva de las cuatro cepas al aumentar la biomasa y la longitud de P. clandestinum.

The indiscriminate use of chemical fertilizers has contributed to the deterioration of the biological, physical and chemical properties of the soil, resulting in the loss of its productive capacity. For this reason, the use of biofertilizers has emerged as a technological alternative. The objective of this research was to develop a suitable liquid fermentation system and encystment for the multiplication of Azotobacter chroococcum AC1 strain, a bacterium employed in a biofertilizer formulation produced at present by CARPOICA, Colombia. Sequential statistical designs were used to determine the conditions in the fermentation system. The interaction between agitation, aeration and pH was evaluated on the viable biomass (CFU/ml) of AC1. In addition, the encystment ability of the strain was evaluated using two encystment agents and the potential plant growth-promoting rhizobacteria (PGPR) activity was assessed by different techniques, such as nitrogen fixation by ARA, phosphate solubilization by the phospho-molybdenum-blue reaction and indolic compound production by colorimetric reaction using the Salkowski reagent. Results showed significant effects (p0.05) on the viable biomass in the three conditions (pH, aeration and agitation) tested individually, in one dual interaction and one tripartite interaction, were demonstrated to have a positive effect on the response variable aeration and agitation. The addition of the two encystment agents evaluated, AE01 and AE02, demonstrated the ability of AC1 to form cysts under stress conditions. Likewise, fermentation and encystment conditions did not affect the biological activities tested.

Phosphorus is limiting for growth of maize plants, and because of that use of fertilizers like rock phosphate has been proposed. However, direct use of rock phosphate is not recommended because of its low availability, so it is necessary to improve it. In this study, a group of diazotrophic bacteria were evaluated as phosphate-solubilizing bacteria, for their production of indolic compounds and for their effects on growth of maize plants. Strains of the genera Azosporillum, Azotobacter, Rhizobium and Klebsiella, were quantitatively evaluated for solubilization of Ca3(PO4)2 and Rock Phosphate as a single source of phosphorous in SRS culture media. Additionally, the phosphatase enzyme activity was quantified at pH 5.0, 7.0 and 8.0 using p-nitrophenyl phosphate, and production of indolic compounds was determined by colorimetric quantification. The effect of inoculation of bacteria on maize was determined in a completely randomized greenhouse experiment where root and shoot dry weights and phosphorus content were assessed. Results showed that strain C50 produced 107.2 mg .L -1 of available-P after 12 days of fermentation, and AC10 strain had the highest phosphatase activity at pH 8 with 12.7 mg of p-nitrophenol mL .h -1 . All strains synthetized indolic compounds, and strain AV5 strain produced the most at 63.03 µg .mL -1 . These diazotrophic bacteria increased plant biomass up to 39 % and accumulation of phosphorus by 10%. Hence, use of diazotrophic phosphate-solubilizing bacteria may represent an alternative technology for fertilization systems in maize