Azotobacter chroococcum as a potentially useful bacterial biofertilizer for cotton (Gossypium hirsutum): Effect in reducing N fertilization / Azotobacter chroococcum como biofertilizante bacteriano potencialmente útil para el algodón (Gossypium hirsutum L.): efecto en la reducción de la fertilización nitrogenada

The aim of this research was to evaluate whether the application of two plant growth-promoting (rhizo)bacteria might reduce nitrogen fertilization doses in cotton. We used strains Azotobacter chroococcum AC1 and AC10 for their proven ability to promote seed germination and cotton growth. These microorganisms were characterized by their plant growth-promoting activities. Then, we conducted a glasshouse study to evaluate the plant growth promoting ability of these strains with reduced doses of urea fertilization in cotton. Results revealed that both strains are capable of fixing nitrogen, solubilizing phosphorus, synthesizing indole compounds and producing hydrolytic enzymes. After 12 weeks, the glasshouse experiment showed that cotton growth was positively influenced due to bacterial inoculation with respect to chemical fertilization. Notably, we observed that microbial inoculation further influenced plant biomass (p<0.05) than nitrogen content. Co-inoculation, interestingly, exhibited a greater beneficial effect on plant growth parameters compared to single inoculation. Moreover, similar results without significant statistical differences were observed among bacterial co-inoculation plus 50% urea and 100% fertilization. These findings suggest that coinoculation of A. chroococcum strains allow to reduce nitrogen fertilization doses up to 50% on cotton growth. Our results showed that inoculation with AC1 and AC10 represents a viable alternative to improve cotton growth while decreasing the N fertilizer dose and allows to alleviate the environmental deterioration related to N pollution.

El objetivo de esta investigación fue evaluar si la aplicación de 2 (rizo)bacterias promotoras del crecimiento vegetal podría reducir la dosis de fertilizante nitrogenado en el cultivo de algodón. Se usaron las cepas Azotobacter chroococcum AC1 y AC10 por su habilidad para promover la germinación de semillas y el crecimiento del algodonero. Estos microorganismos fueron caracterizados sobre la base de sus actividades de promoción del crecimiento vegetal. Luego se realizó un estudio de invernadero con plantas de algodón para evaluar la capacidad de promoción del crecimiento vegetal de dichas cepas con dosis reducidas de urea. Los resultados revelaron que ambas cepas son capaces de fijar nitrógeno, solubilizar fósforo, sintetizar compuestos indólicos y producir enzimas hidrolíticas. Después de 12 semanas, el experimento de invernadero permitió observar que el crecimiento del algodón fue influido positivamente por la inoculación bacteriana con respecto a la fertilización química. En particular, se evidenció que la inoculación microbiana impactó más en la biomasa vegetal (p<0,05) que en el contenido de nitrógeno. Curiosamente, la coinoculación exhibió un mayor efecto positivo sobre los parámetros de crecimiento en comparación con la inoculación simple. Además, se observaron resultados similares, sin diferencias estadísticamente significativas, entre la coinoculación bacteriana más del 50% de urea y el 100% de fertilización. Estos hallazgos indican que la coinoculación de las cepas de A. chroococcum AC1 y AC10 permitiría reducir las dosis de fertilización nitrogenada del cultivo de arroz en hasta el 50% y aliviar, de esta manera, el deterioro ambiental relacionado con la contaminación por N.

Compatibility of Azospirillum brasilense and Pseudomonas fluorescens in growth promotion of groundnut ( Arachis hypogea L)

ABSTRACT We attempted to study the compatibility among plant beneficial bacteria in the culture level by growing them near in the nutrient agar plates. Among all the bacteria tested, Rhizobium was found to inhibit the growth of other bacteria. From the compatible group of PGPR, we have selected one biofertilizer (Azospirillum brasilense strain TNAU) and one biocontrol agent (Pseudomonas fluorescens strain PF1) for further studies in the pot culture. We have also developed a bioformulation which is talc powder based, for individual bacteria and mixed culture. This formulation was used as seed treatment, soil application, seedling root dip and foliar spray in groundnut crop in vitro germination conditions. A. brasilense was found to enhance the tap root growth and P. fluorescens, the lateral root growth. The other growth parameters like shoot growth, number of leaves were enhanced by the combination of both of the bacteria than their individual formulations. Among the method of application tested in our study, soil application was found to be the best in yielding better results of plant growth promotion.

Uso de Leersia hexandra (Poaceae) en la fitorremediación de suelos contaminados con petróleo fresco e intemperizado / Use of Leersia hexandra (Poaceae) for soil phytoremediation in soils contaminated with fresh and weathered oil

ResumenLa industria petrolera ha generado derrames crónicos de petróleo y su acumulación en Gleysoles en zonas anegadas en el estado de Tabasco, en el sureste de México. El anegamiento es un factor que limita el uso de tecnologías de remediación por el alto costo y los bajos niveles de degradación del petróleo, sin embargo, Leersia hexandra Sw. es un pasto que crece en estas zonas contaminadas con petróleo intemperizado. El objetivo del estudio fue evaluar la densidad de bacterias, producción de biomasa vegetal y fitorremediación de L.hexandra en suelo contaminado con petróleos fresco e intemperizado, bajo condiciones experimentales de anegamiento. Se realizaron dos experimentos (E1 y E2) en un túnel de plástico. El E1 se basó en ocho dosis: 6 000, 10 000, 30 000, 60 000, 90 000, 120 000, 150 000 y 180 000 mg kg-1 base seca (b.s.) de hidrocarburos totales de petróleo fresco (HTPF), y en el E2 se evaluaron cinco dosis: 14 173, 28 400, 50 598, 75 492 y 112 142 mg kg-1 b. s. de hidrocarburos totales de petróleo intemperizado (HTPI), con ocho repeticiones en cada experimento, además se utilizó un testigo con 2 607 mg kg-1 b. s. de HTP de origen biogénico. Las variables evaluadas a los tres y seis meses fueron 1) densidad microbiana de las bacterias fijadoras de nitrógeno de vida libre totales (BFN), del grupo Azospirillum (AZP) y Azotobacter (AZT), por cuenta viable en placa seriada; 2) producción de materia seca total (MS), se cuantificó por el peso seco por gravimetría, y 3) el porcentaje de descontaminación de los hidrocarburos (DSC) por extracción en equipo soxhlet. En suelos con HTPF, la población de BFN, AZP y AZT se estimuló hasta cinco veces más que el tratamiento testigo a los tres y seis meses; sin embargo, concentraciones de 150 000 y 180 000 mg kg-1 b. s. inhibieron entre un 70 y 89 % la densidad bacteriana. A su vez, en suelos con PI, la inhibición se registró hasta en un 90 %, a excepción del tratamiento con 14 173 mg kg-1 b. s., el cual estimuló las BNF y AZT en 2 y 0.10 veces más que testigo, respectivamente. La producción de MS fue continua en los experimentos hasta los seis meses, con valores de 63 y 89 g en PF y PI, respectivamente; sin diferencias significativas con el testigo (p ≤ 0.05). El DSC alcanzó valores del 66 % al 87 % en HTPF como HTPI a los seis meses, respectivamente. Estos resultados demuestran la habilidad del L. hexandra para desarrollar una rizósfera con alta densidad de BFN, producir biomasa vegetal y fitorremediar Gleysoles con petróleo fresco e intemperizado en ambientes tropicales inundados.

Abstract:The oil industry has generated chronic oil spills and their accumulation in wetlands of the state of Tabasco, in Southeastern Mexico. Waterlogging is a factor that limits the use of remediation technologies because of its high cost and low levels of oil degradation. However, Leersia hexandra is a grass that grows in these contaminated areas with weathered oil. The aim of the study was to evaluate the bacteria density, plant biomass production and phytoremediation of L. hexandra in contaminated soil. For this, two experiments in plastic tunnel were performed with fresh (E1) and weathered petroleum (E2) under waterlogging experimental conditions. The E1 was based on eight doses: 6 000, 10 000, 30 000, 60 000, 90 000, 120 000, 150 000 and 180 000 mg.kg-1 dry basis (d. b.) of total petroleum hydrocarbons fresh (TPH-F), and the E2, that evaluated five doses: 14 173, 28 400, 50 598, 75 492 and 112 142 mg. kg-1 d. b. of total petroleum hydrocarbons weathered (TPH-W); a control treatment with 2 607 mg.kg-1 d. b. was used. Each experiment, with eight replicates per treatment, evaluated after three and six months: a) microbial density of total free-living nitrogen-fixing bacteria (NFB) of Azospirillum (AZP) and Azotobacter group (AZT), for viable count in serial plate; b) dry matter production (DMP), quantified gravimetrically as dry weight of L. hexandra; and c) the decontamination percentage of hydrocarbons (PDH) by Soxhlet extraction. In soil with TPH-F, the NFB, AZP y AZT populations were stimulated five times more than the control both at the three and six months; however, concentrations of 150 000 and 180 000 mg.kg-1 d. b. inhibited the bacterial density between 70 and 89 %. Likewise, in soil with TPH-W, the FNB, AZP and AZT inhibitions were 90 %, with the exception of the 14 173 mg.kg-1 d. b. treatment, which stimulated the NFB and AZT in 2 and 0.10 times more than the control, respectively. The DMP was continued at the six months in the experiments, with values of 63 and 89 g in fresh and weathered petroleum, respectively; had no significant differences with the control (p≤0.05). The PDH reached values of 66 to 87 % both TPH-F and TPH-W at six months, respectively. These results demonstrated the ability the L. hexandra rhizosphere to stimulate the high NFB density, vegetal biomass production and phytoremediation of contaminated soils (with fresh and weathered petroleum), in a tropical waterlogging environment. Rev. Biol. Trop. 65 (1): 21-30. Epub 2017 March 01.

The use of lacZ marker in enumeration of Azotobacter chroococcum in carrier based inoculants

A transconjugant of Azotobacter chroococcum Mac 27 tagged with lac Z(A. chroococcum Mac27 L) was found to possess high levels of β-galactosidase activity constitutively.Further, the lac Z marker was found to be stably integrated into the chromosome of the A. chroococcum Mac 27 and did not have any adverse effect on growth, nitrogen fixation and excretion of ammonia. A quick method to determine the viable cell number in broth culture and carrier based inoculants has been developed on the basis of β-galactosidase assay. It was found that there was a direct relationship between the number of cell as determined by standard plate count and intensity of colour that developed upon degradation of ONPG due to β-galactosidase activity .The method was found to be sensitive enough to determine 1.7 x 10(6) CFU mL-1 in broth culture as well as carrier based Azotobacter inoculants. Further, it was observed that when A. chroococcum Mac27 L was inoculated on Brassica campestris, it could be detected in the presence of other bacteria capable of growing on Burks agar medium containing X-gal on the basis of lac Z genetic marker.

Evaluación de la estabilidad de Trichoderma sp. y Azotobacter sp. conservados por diferentes métodos / Evaluation of the stability of Trichoderma sp. and Azotobacter sp. preserved by different methods

En el Laboratorio de Microbiología Aplicada de la Universidad Francisco de Paula Santander (Cúcuta, Colombia) constantemente ingresan al Banco de Cepas, cultivos microbianos de interés biotecnológico, especialmente del sector agrícola, los cuales son utilizados en las actividades de docencia e investigación. Por esta razón, existe el interés de mantener viables a través del tiempo estos cultivos microbianos, para lo cual se realizó en esta investigación, la evaluación de la estabilidad de los aislados de Trichoderma sp. y Azotobacter sp., utilizando las técnicas de conservación en viales con solución salina estéril (0,85% NaCl) en refrigeración (4°C) y temperatura ambiente (30°C), suelo estéril en refrigeración (4°C), comparados con la metodología de repiques sucesivos como tratamiento control. Los resultados no mostraron diferencias significativas según el Test de Duncan (P≤0,05) en la tasa de supervivencia microbiana entre los métodos de conservación. Sin embargo, se observó en las técnicas de viales con solución salina estéril y suelo estéril mantenidos en refrigeración, mayor estabilidad en la concentración celular durante los cuatros meses de evaluación, sin registrar contaminación en los cultivos. Así mismo, se registró un óptimo crecimiento macroscópico de los cultivos microbianos y sus características microscópicas se mantuvieron estables en estos dos métodos de conservación. Por esta razón, se seleccionaron como técnicas de conservación de estos microorganismos, teniendo en cuenta además, ventajas como la facilidad de la técnica, disponibilidad de equipos, materiales y personal con los que se cuenta en el laboratorio.

In the Laboratory of Applied Microbiology of the University Francisco de Paula Santander (Cúcuta, Colombia), strains of microbial cultures of biotechnological interest are constantly entering, particularly from the agricultural sector, which are used for teaching and research activities. There is an interest in keeping them viable over time, which was the main reason of this research: To evaluate the stability of the isolates of Trichoderma sp. and Azotobacter sp., using different conservation techniques: vials with saline solution (0,85% NaCl) in two temperature conditions: refrigeration (4°C) and room temperature (30°C), sterile soil under refrigeration (4°C), and successive peals methodology as the control treatment. Results showed no significant differences according to Duncan test (P ≤ 0.05) in the microbial survival ratio between preservation methods. However, a greater stability in cell concentration was observed with the techniques of vials with sterile saline solution and sterile soil -both under refrigeration-, during the four-month evaluation, with no register of contamination on cultures. Also, there was an optimum growth of microbial cultures macroscopically and the microscopic characteristics were stable in these two methods of preservation for both strains. For this reason, we suggest these conservation techniques for the two microorganisms, considering further advantages such as ease of technology, availability of equipment, materials and personnel available in the laboratory.

[Response of quantity and quality yield of valerian [Valeriana officinalis L.] to application of phosphorous bio/chemical fertilizers]

Since phosphorus is the vital nutrient for plants, the evaluation of phosphorous fertilizers effects on yield and quality of valerian is necessary. In this research the effect of phosphorus solubilization bacterias as bio-fertilizers and phosphorous chemical fertilizer on quantity and quality yield of valerian was studied. This study conducted in the research farm of Institute of Medicinal Plants [IMP] through a factorial experiment based on a randomized complete blocks design with three replications. Factors of experiment included the inoculation bio-fertilizers factor in four levels: Azotobacter, Pseudomonas, mix of both and control [without inoculation], and also the phosphorous chemical fertilizer factor as super phosphate triple in three levels: 0, 50 and 100 kg ha[-1]. The bio-fertilizers using had a significant effect [p

Production and partial characterization of serine and metallo peptidases secreted by Aspergillus fumigatus Fresenius in submerged and solid state fermentation

Enzyme production varies in different fermentation systems. Enzyme expression in different fermentation systems yields important information for improving our understanding of enzymatic production induction. Comparative studies between solid-state fermentation (SSF) using agro-industrial waste wheat bran and submerged fermentation (SmF) using synthetic media were carried out to determinate the best parameters for peptidase production by the fungus Aspergillus fumigatus Fresen. Variables tested include: the concentration of carbon and protein nitrogen sources, the size of the inoculum, the pH of the media, temperature, and the length of the fermentation process. The best peptidase production during SSF was obtained after 96 hours using wheat bran at 30 ºC with an inoculum of 1 x 10(6) spores and yielded 1500 active units (UµmL). The best peptidase production using SmF was obtained after periods of 72 and 96 hours of fermentation in media containing 0.5% and 0.25% of casein, respectively, at a pH of 6.0 and at 30 ºC and yielded 40 UµmL. We also found examples of catabolite repression of peptidase production under SmF conditions. Biochemical characterization of the peptidases produced by both fermentative processes showed optimum activity at pH 8.0 and 50 ºC, and also showed that their proteolytic activity is modulated by surfactants. The enzymatic inhibition profile using phenylmethylsulfonyl fluoride (PMSF) in SmF and SSF indicated that both fermentative processes produced a serine peptidase. Additionally, the inhibitory effect of the ethylene-diaminetetraacetic acid (EDTA) chelating agent on the peptidase produced by SmF indicated that this fermentative process also produced a metallopeptidase.

Influencia de la materia orgánica y Azotobacter nigricans en un cultivo de Stevia rebaudiana B / Influência da matéria orgânica e Azotobacter nigricans em uma cultura de Stevia rebaudiana B / Influence of organic matter and Azotobacter nigricans on a Stevia rebaudiana B. plantation

Stevia rebaudiana es una planta que acumula un compuesto edulcorante no calórico conocido como esteviosido. Su cultivo tradicionalmente se fertiliza con compuestos químicos que causan daño al ecosistema, obligando a buscar alternativas orgánicas para mitigar este daño. Objetivo. Evaluar el efecto de la materia orgánica y un biofertilizante con base en Azotobacter nigricans en un cultivo de Stevia rebaudiana ubicado en el departamento de Meta, Colombia. Materiales y métodos. Se establecieron cinco tratamientos: T1 y T2 Incorporando al cultivo compost de residuos domiciliarios en concentraciones de 15 y 30 ton ha-1 respectivamente; T3 y T4 con las mismas concentraciones de compost e inoculando el biofertilizante y T5 solamente con el biofertilizante. El control correspondió a las condiciones normales de cultivo con compost Bokashi. El crecimiento vegetal se evaluó por determinación de biomasa en peso seco, producción y área foliar. El análisis fisicoquímico de suelo incluyó: porcentaje carbono orgánico, humedad y pH. Resultados. La inoculación del biofertilizante produjo un incremento en la velocidad de mineralización del compost, con %CO final de 4,85 entre los 90 y 180 días. Se presentaron diferencias estadísticamente significativas (p< 0,05) entre la producción de biomasa de T2 (1538 kg ha-1) y el control (477 kg ha-1). Respecto al contenido de sólidos solubles T1, T3 y T4 presentaron los más altos valores de °Brix (12,4: 12,35 y 12,15). Conclusiones. La concentración 30 ton ha-1 presentó los mejores rendimientos de biomasa y la aplicación del biofertilizante mostró una correlación positiva con la mineralización del compost y síntesis de glucósidos...

Influence of organic matter and Azotobacter nigricans on a Stevia rebaudiana B. plantation Stevia rebaudiana is a plant that accumulates a non-caloric sweetener compound known as stevioside. This crop is traditionally fertilized with chemicals that are harmful for the ecosystem, forcing to find organic alternatives to mitigate this damage. Objective. To study the effect of organic matter and an Azotobacter nigricans-based bio-fertilizer on a Stevia rebaudiana plantation grown in acidic soil in the Department of Meta, Colombia. Materials and methods. Five treatments were established: T1 and T2 with the application of home organic waste compost at concentrations of 15 and 30 ton ha-1. T3 and T4 with the same compost concentrations and inoculating the A. nigricans bio-fertilizer. T5 contained the bio-fertilizer alone. The control consisted of the application of the Bokashi compost under the usual conditions of cultivation. Plant growth was assessed by biomass increase measured as dry weight, production, and leaf area. The physicochemical analysis of soil included: percentage of organic carbon, water content, and pH. Results. The inoculation of the bio-fertilizer produced an increase in the rate of mineralization of compost, reaching a final 4.85% of OC between 90 and 180 days after inoculation. There were significant (p< 0.05) differences between biomass production with T2 (1,538 kg ha-1) and the control (477 kg ha-1). Regarding the soluble solid content, T1, T3 and T4 showed the highest °Brix values (12.4, 12.35 and 12.15, respectively). Conclusions. The concentration of 30 ton ha-1 produced the highest biomass production and the application of the biofertilizer showed a positive correlation with compost mineralization and glucoside synthesis...

Influência da matéria orgânica e Azotobacter nigricans em uma cultura de Stevia rebaudiana B. Stevia rebaudiana é uma planta que acumula um composto edulcorante não-calórico conhecido como esteviosídeo. Sua cultura é tradicionalmente fertilizada com produtos químicos que danificam o ecossistema, forçando a procura de alternativas orgânicas para reduzir esse dano. Objetivo. Avaliar o efeito da matéria orgânica e um biofertilizante com base em Azotobacter nigricans numa cultura de Stevia rebaudiana localizada no departamento de Meta, na Colômbia. Materiais e métodos. Cinco tratamentos foram estabelecidos: T1 e T2 incorporando ao cultivo composto de lixo doméstico em concentrações de 15 e 30 t ha-1, respectivamente; T3 e T4 com as mesmas concentrações do composto e inoculando o biofertilizante e T5 apenas com o biofertilizante. O controle correspondeu a condições normais de cultivo com composto Bokashi. O crescimento das plantas foi avaliado por estimação da biomassa em peso seco, da produção e da área foliar. A análise físico-química do solo inclui o seguinte: porcentagem de carbono orgânico, umidade e pH. Resultados. A inoculação do biofertilizante produz um aumento na taxa de mineralização do composto, com CO final de 4,85% entre 90 e 180 dias. Diferenças estatisticamente significativa (p <0,05) foram observadas entre a produção de biomassa de T2 (1538 kg ha-1) e o controle (477 kg ha-1). Com relação ao conteúdo de sólidos solúveis T1, T3 e T4 tiveram os maiores valores de °Brix (12,4: 12,35 e 12,15). Conclusões. A concentração de 30 ton ha-1 apresentou o melhor rendimento de biomassa e a aplicação do biofertilizante mostrou uma correlação positiva com a mineralização do composto e a síntese de glicosídeos...

Efecto biofertilizante del preparado: residuos vegetales - bacteria nativa diazótrofa, sobre las variables biométricas en plántulas de Rhapanus sativus / Biofertilizer effect of the prepared from vegetales wastes - diazotroph native bacterium on biometrics variables of Rhapanus sativus seedlings

El uso de bioinoculantes a base de microorganismos con potencial biofertilizante representa una alternativa económicamente viable y de producción limpia para el sector agrícola. El objetivo del presente trabajo fue evaluar el efecto biofertilizante de un preparado elaborado con residuos sólidos vegetales (RSV) procedentes del mercado y la bacteria nativa diazótrofa Azotobacter A15M2G. Se elaboraron biopreparados utilizando diferentes concentraciones de bacteria (106, 107 y 108 UFC) en un medio de cultivo obtenido a partir del 25% p/v de cada uno de los siguientes RSV: Brassica oleracea (repollo), Lactuca sativa (lechuga) y Allium fistulosum (cebollín). Los biopreparados fueron evaluados en plantas de rábano (Rhapanus sativus) en invernadero, utilizando un diseño estadístico completamente al azar de 5 tratamientos con 3 repeticiones: T1, control; T2, semillas pregerminadas tratadas con RSV al 25% p/v; T3, semillas pregerminadas con bioinoculante de 106 UFC; T4, semillas pregerminadas con bioinoculante de 107 UFC; T5, semillas pregerminadas con bioinoculante de 108 UFC. Se evaluó: número de hojas, área foliar, longitud de la planta, longitud de la raíz y peso seco de toda la planta (ensayos por triplicado). Se observó un incremento altamente significativo en peso seco para T5 (0,88 g) y T4 (1,10 g); y diferencias significativas en el área foliar, para los mismos tratamientos, con un valor superior a 2000 cm2. El biopreparado con bacterias nativas y RSV mejoró el crecimiento y desarrollo de las plantas de rábano, pudiéndose dar un valor agregado a estos residuos y de esta manera obtener un biofertilizante potencialmente utilizable en otros cultivos.

The use of bioinoculantes from microorganisms with biofertilizer potential, represents an economically viable alternative and of clean production for the agricultural sector. The aim of this study was to evaluate the effect of biofertilizer preparation obtained from vegetable solid waste (RSV) of the market and the native bacteria Azotobacter A15M2G diazotroph.Biological cultures were prepared using different inoculum concentrations, 106, 107 y 108 UFC in a culture medium obtained from 25% w / v of each of the following substrates: Brassica oleracea (cabbage), Lactuca sativa (lettuce) and Allium fistulosum (chives). The microbial inoculants were evaluated in radish plants (Rhapanus sativus) in greenhouse using a completely randomized design of 5 treatments with 3 replicates: T1, pre-germinated seeds without any treatment; T2, pre-germinated seeds treated with the dye waste vegetables 25% w / v; T3, pre-germinated seeds treated with bacterial concentration bioinoculants to 106 UFC; T4, pre-germinated seeds treated with bacterial concentration bioinoculants to 107 UFC, and T5, pre-germinated seeds treated with bacterial concentration bioinoculants to 108 UFC. Assessed variables were: number of leaves, leaf area, plant length, root length and dry weight of the entire plant (all assays in triplicate). The results showed a highly significant increase in dry weight, for T5 (0.88 g) and T4(1.10 g); and significant differences in leaf area for the same treatments, with a value greater than 2000 cm2, compared to others. The biopreparado from native bacteria and RSV improved the growth and development of the radish plants, being able to give a added value to these residues and to obtain a potentially usable biofertilizer in other cultures.

Growth and yield responses of tomato [lycopersicon esculentum] grown under soilless cultivation to application of azospirillum, azotobacter and nitrogen

THIS STUDY was undertaken to investigate the possibility of using Azospirillum bras/lease and Azotobacter chroococcum applied with inorganic nitrogen to enhance tomato [Lycopersicon escnlentum, Mill] production in soilless systems. The effect of inoculation with A. brasilense and / or Azoto. chroococcum with the application of 75% [of the recommended dose] of N[2] on certain tomato growth parameters [shoot height, number of leaves, and fresh and dry weights of plants] after 30, 60 and 90 days of transplanting were determined. In addition, total yield, mean fruit weight and number of fruits per plant were evaluated in plants inoculated with the two bacteria [separately or in combination] with 75% N[2], and plants inoculated with 75% N[2] [control 1] and 100% N[2] [control 2], Plants inoculated with a mixed inoculum of A. bras/lease and Azoto. chroococcum with 75% N[2] gave the highest values of the growth parameters tested while plants inoculated with 75% N[2] [control 1] gave the lowest. The same treatment also resulted in the highest tomato yield, mean weight and number of fruits per plant followed by plants treated with A. brasilense and 75% N[2]. The responses to the tested biofertilizers on nitrogen, phosphorous and potassium [NPK] uptake, population of diazotrophic bacteria and nitrogenase and dehydrogenase activity of the tested bacteria on the rhizosphere of tomato showed that the mixed inoculum of A. brasilense and Azoto. chroococcum with 75% N[2] gave the highest NPK uptake levels after 30, 60 and 90 days of transplanting. The same treatment gave the highest population of diazotrophic bacteria and dehydrogenase and nitrogenase activity of the tested bacteria compared with the plants inoculated with a single organism or controls. In contrast, control 1 gave the least values among all treatments. This study shows that inoculation of tomato plants with Azospirillum brasilense and Azolobacter chroococcum have a significantly beneficial effect on tomato yield under soilless cultivation

Fatty acids in an estuarine mangrove ecosystem

Fatty acids have been successfully used to trace the transfer of organic matter in coastal and estuarine food webs. To delineate these web connections, fatty acid profiles were analyzed in species of microbes (Azotobacter vinelandii, and Lactobacillus xylosus), prawns (Metapenaeus monoceros and Macrobrachium rosenbergii) and finfish (Mugil cephalus), that are associated with decomposing leaves of two mangrove species, Rhizophora apiculata and Avicennia marina. The fatty acids, except long chain fatty acids, exhibit changes during decomposition of mangrove leaves with a reduction of saturated fatty acids and an increase of monounsaturated fatty acids. The branched fatty acids are absent in undecomposed mangrove leaves, but present significantly in the decomposed leaves and in prawns and finfish, representing an important source for them. This revealed that the microbes are dominant producers that contribute significantly to the fishes and prawns in the mangrove ecosystem. This work has proved the fatty acid biomarkers as an effective tool for identifying the trophic interactions among dominant producers and consumers in this mangrove. Rev. Biol. Trop. 58 (2): 577-587. Epub 2010 June 02.

Los ácidos grasos se han utilizado con éxito para estudiar la transferencia de materia orgánica en las redes alimentarias costeras y estuarinas. Para delinear las interacciones tróficas en las redes, se analizaron perfiles de ácidos grasos en las especies de microbios (Azotobacter vinelandii y Lactobacillus xylosus), camarones (Metapenaeus monoceros y Macrobrachium rosenbergii) y peces (Mugil cephalus), que están asociadas con la descomposición de las hojas de dos especies de mangle, Rhizophora apiculata y Avicennia marina. Los ácidos grasos, con excepción de los de cadena larga, exhiben cambios durante la descomposición de las hojas de mangle, con una reducción de los ácidos grasos saturados y un aumento de los monoinsaturados. Los ácidos grasos ramificados están ausentes en las hojas de mangle sin descomponer, pero presentes de manera significativa en las hojas descompuestas, en camarones y peces, representando una fuente importante para ellos. Esto revela que los microbios son productores dominantes que contribuyen significativamente con los peces y camarones en el ecosistema de manglar. Este trabajo demuestra que los marcadores biológicos de los ácidos grasos son una herramienta eficaz para la identificación de las interacciones tróficas entre los productores dominantes y consumidores en este manglar.

Effect of carbon source and precursors on the production of plant growth regulators by azotobacter chroococcum [R19] and bacillus megaterium var. phosphaticum [R44]

An experiment was carried out to examine the effect of carbon source, tryptophan and adenine concentrations on the production of growth regulators by A. chroococcum [R19] and B. megaterium var. phosphaticum [R44]. Mannitol and glucose were the best carbon sources for the production of plant growth regulators [PGRs] by A. chroococcum and B. megaterium var. phosphaticum, respectively. A. chroococcum produced higher amounts of zeatin and kinetin compared to those produced by B. megaterium var. phosphaticum while B. megaterium var. phosphaticum produced higher amounts of [9R] benzyl adenine and [9G] bcnzyl adenine compared with those produced by A. chroococcum. Production of auxins, gibberellic acid [GA[3]] and cytokinins was increased with increasing tryptophan concentration. The highest amounts of PGRs produced by the two strains were obtained with tryptophan at 1000 micro M. Highest amounts of PGRs produced by A. chroococcum and B. megaterium var. phosphaticum at 10 and 100 micro M of adenine, respectively. Also, the produced amounts of gibberellic acid and cytokinin than that produced by B. megaterium var. phosphaticum. Generally, obtained data showed that the application of the optimal conditions together gave highest amounts of PGRs as compared with the other individual factors. This result is logic and was anticipated

Producción de un biofertilizante a partir de un aislamiento de Azotobacter nigricans obtenido en un cultivo de Stevia rebaudiana Bert / Produção de um bio-fertilizante a partir do isolamento de Azotobacter nigricans obtido num cultivo de Stevia rebaudiana Bert. rato / Bio-fertilizer production from an isolate of Azotobacter nigricans obtained from a plantation of Stevia rebaudiana Bert

Realizar un aislamiento de bacterias fijadoras de nitrógeno para emplearlas en un programa de fertilización bajo un esquema de agricultura orgánica. Materiales y métodos. El aislamiento de bacterias fijadoras de nitrógeno se realizó en medio Ashby-benzoato a partir del suelo de un cultivo de Stevia rebaudiana Bert. Los aislamientos identificados como Azotobacter nigricans fueron evaluados mediante una cinética de crecimiento y la cepa con mayor velocidad se utilizó para la elaboración de un biofertilizante por fermentación discontinua La evaluación preliminar del biofertilizante se realizó mediante su inoculación en 3 eras de un cultivo de S. rebaudiana Bert. y el rendimiento se determinó con base en la producción de biomasa y concentración de glucósidos. Resultados. Dos aislamientos (A5 y A6) fueron identificados como A. nigricans con base en la caracterización fenotípica y genotípica. El aislamiento A5 se seleccionó para la elaboración del biofertilizante debido a que presentó mejor estabilidad, pigmentación, mayor velocidad de crecimiento 0,1405 h-1 fase exponencial de 18 horas y una producción de AIA promedio de 38,4 mg/ml a las 150 horas. El biofertilizante se obtuvo en medio leche con una concentración celular de 4x1012 UFC/ml. Conclusiones. La evaluación preliminar en campo mostró una correlación positiva entre el aumento de la concentración de glucósidos en las hojas de S. rebaudiana y una mayor producción de biomasa en respuesta a la aplicación del biofertilizante...

Bio-fertilizer production from an isolate of Azotobacter nigricans obtained from a plantation of Stevia rebaudiana Bert. Objective.To isolate nitrogen fixing bacteria to be used in a fertilization regime of an organic agriculture program. Materials and methods. The isolation of nitrogen fixing bacteria was done in an Ashby-benzoate medium from soil of a Stevia rebaudiana plantation. Isolates identified as Azotobacter nigricans were evaluated by their growth kinetics and the strain with the fastest growth was used for the production of a biofertilizer by discontinuous fermentation. The preliminary evaluation of the biofertilizer was done by its inoculation into three ridges of a plantation of S. rebaudiana and yield determination was based upon biomass production and glycoside concentration. Results. Two isolates (A5 and A6) were identified as A. nigricans based on their phenotypic and genotypic characterization. Isolate A5 was selected for preparing the biofertilizer because it showed a better stability, pigmentation, a faster growth rate (0.1405 h-1 exponential phase of 18 hours) and an average IAA production of 38.4 mg/ml after 150 hours. The bio-fertilizer was obtained in milk medium with a cell concentration of 4x1012 CFU/ml. Conclusions. The preliminary field evaluation showed a positive correlation between the increase of the glycoside concentration in the leaves of S. rebaudiana and a higher production of biomass in response to the bio-fertilizer application...

Produção de um bio-fertilizante a partir do isolamento de Azotobacter nigricans obtido num cultivo de Stevia rebaudiana Bert.Objetivo: Realizar um isolamento de bactérias fixadoras de nitrogênio para empregá-las num programa de fertilização sob um esquema deagricultura orgânica. Materiais e métodos: O isolamento de bactérias fixadoras de nitrogênio realizou-se no meio Ashby-benzoato a partirdo solo de um cultivo de Stevia rebaudiana Bert. Os isolamentos identificados como Azotobacter nigricans foram avaliados mediante umacinética de crescimento e a cepa com maior velocidade utilizou-se para a elaboração de um bio-fertilizante por fermentação descontínua. Aavaliação preliminar do bio-fertilizante realizou-se mediante sua inoculação em 3 eras de um cultivo de S. rebaudiana Bert., e o rendimento determinou-se com base na produção de biomassa e concentração de glucosideos. Resultados: Dois isolamentos (A5 e A6) foram identificados como A. nigricans com base na caracterização fenotípica e genotípica. O isolamento A5 selecionou-se para a elaboração do bio-fertilizante por apresentar melhor estabilidade, pigmentação, maior velocidade de crescimento (0,1405 h-1 fase exponencial de 18 horas) e uma produção de AIA media de 38,4? mg/ml as 150 horas. O bio-fertilizante obteve-se no meio leite com uma concentração celular de 4x1012 UFC/ml. Conclusões. A avaliação preliminar no campo apresentou uma correlação positiva entre o aumento da concentração deglucosideos nas folhas de S. rebaudiana e uma maior produção de biomassa como resposta à aplicação do bio-fertilizante...

Bacterias fijadoras asimbióticas de nitrógeno de la zona agrícola de San Carlos. Córdoba, Colombia / Non-symbiotic bacterial diazotrophs from of agricultural crops of San Carlos. Córdoba, Colombia

Se aislaron bacterias diazotróficas de los géneros Azotobacter sp y Azospirrillum sp a partir de la rizosfera de cultivos de plátano, pastos, maíz y de rastrojos (zonas sin cultivar) en el municipio de San Carlos (Valle del Sinú medio en el departamento de Córdoba, Colombia). Las poblaciones microbianas se identificaron mediante pruebas bioquímicas y observación macroscópica y microscópica con tinción de Gram en diferentes medios de cultivo: a) Burk´s, Asbhy y Jensen´s, (género Azotobacter sp), y b) Burk´s, nfb y rojo congo (género Azospirillum sp). El objetivo del presente trabajo fue determinar la producción del ión amonio a partir de los géneros bacterianos aislados;la cuantificación del ión amonio fue llevada a cabo por el método colorimétrico de Berthelot (fenol-hipoclorito) empleando un espectrofotómetro Perkin-Elmer Lamba 11 uv-Vis; la técnica fue modificada y estandarizada de acuerdo con las condiciones del equipo. Como resultado seobtuvieron 14 aislados que produjeron concentraciones de 0,9 hasta 5,2 mg/l, siendo los más destacados para el género Azotobacter sp, A16PG y A26M1P (5,1545 y 5,1743 mg/l de amonio, respectivamente), y para el género Azospirrillum spp, A5M1G (4,6741 mg/l de amonio). La fijación biológica del nitrógeno (fbn) por bacterias diazotróficas ha contribuido a incrementar el rendimiento en las cosechas, reduciendo la necesidad de fertilizantes nitrogenados y la emisión de gases tóxicos como el N2O, obteniendo beneficios económicos y ambientales en las granjas.

Growth of Avicennia marina and Ceriops decandra seedlings inoculated with halophilic azotobacters.

Inoculation of azotobacter has significant positive effects on the growth characteristics and pigments in mangrove seedlings of Avicennia marina and Ceriops decandra. The bacterial inoculation significantly increased the root dry biomass at the maximum of 75.8% at 30 gl(-1) salinity in Ceriops decandra. But in Avicennia marina, the shoot dry biomass was increased significantly at the maximum of 56.12% at 30 gl(-1) salinity in general, the Azotobacter beijerinkii improved the growth characteristics better in both species of mangroves preferably at higher salinity levels in A. marina and at a range of salinity in C. decandra. The results recommend this forraising vigorous seedlings under nursery conditions.

Growth-associated production and characterization of poly (3-hydroxybutyric acid) of Azotobacter beijerinckii DAR-102.

Production of poly (3-hydroxybutyric acid) [P(3HB)] by Azotobacter beijerinckii DAR-102 isolated in this laboratory has been optimized under batch-culture. The accumulatad polymer attained 58% of cell dry mass during mid-stationary phase with an yield of 0.58 g/l when grown in nitrogen-free medium. The optimum concentration of glucose and fructose for P(3HB) production was 3% (w/v) and 2% (w/v) respectively while that of casamino acid and tryptose was 0.1% (w/v). Phosphate at a concentration suboptimal for growth and limitation of oxygen in the medium favoured P(3HB) accumulation. The production of P(3HB) was maximum with an inoculum dose of 4% (v/v). The accumulated polymer was isolated by direct chloroform extraction of the dry cell mass and purified by precipitation with diethyl ether. The purified polymer has been characterized in terms of its solubility properties, melting temperature, and UV-, IR- and NMR-spectroscopic analyses.

Analogue-resistant mutants of Azotobacter chroococcum derepressed for nitrogenase activity and early ammonia excretion having potential as inoculants for cereal crops.

Spontaneous mutants resistant to methionine sulfoximine (Msx), methyl alanine (Mal) and methyl ammonium chloride (Mac) were derived from A. chroococcum strain A103. Msx and Mal-resistant mutants expressed 1.73 to 10.98% of the fully derepressed nitrogenase activity when grown in Burk's medium containing ammonium acetate. Mac-resistant mutants did not express nitrogenase activity in ammonium acetate supplemented medium. The mutants excreted ammonia even after 2 days of growth and some mutants excreted more ammonia as compared to the parent. Selected mutants were inoculated on wheat (Triticum aestivum) and barley (Hordeum vulgare) under field conditions. Majority of the derepressed mutants increased grain yield of wheat and barley varying from 1.2 to 33.3%. However, host-dependent effects on grain yield were observed with different mutants. Two mutants, Mal 27 and Mac 19 showed significant increase in grain yields of both the crops. The results suggest that metabolic analogue-resistant mutants of Azotobacter have potential for use as a biofertilizer for cereal crops.

Seed borne nature of Azotobacter chroococcum in chilli (Capsicum annum) and its role in seed germination and plant growth.

Investigations were carried out on seed borne nature of Azotobacter chroococcum in chilli and their role in plant growth at the Department of Agriculture Microbiology, University of Agricultural Sciences, Dharwad. Azotobacter chroococcum were isolated from both surface sterilized and unsterilized seeds of 14 varieties of chilli at different stages of fruit maturity. Inoculation with these strains increased the seed germination, root, shoot length and total dry matter content of chilli plants significantly in both sterilized and unsterilized soil., the results of which are presented.

Isolation and characterization of azotobacter strains from brazilian soils

Thirty-eight Azotobacter spp strains isolated from Brasilian soils were characterized for the foollowing properties : utilization of different carbon sources, diffusible pigment production, melanin production, sodium-dependent growth, siderophore production, streptomycin resistance and antibiotic-like substances production. Four strains were identified : one as A. armeniacus and three as A. paspali.

Effect of malathion on certain beneficial microbial activities in Egyptian soils

Malathion was incorporated into Azotobacter chrococcum growth medium with different concentrations up to 600 ppm. The increase in the insecticide concentrations gradually inhibited microbial growth and nitrogen fixation. Meanwhile, the fixed nitrogen continued to increase, for each concentration throughout the experimental period [30 days]. On the other hand, low doses of malathion [100 and 200 ppm] increased both viable count of Bacillus megaterium and its capability of dissolving rock phosphate all over the incubation periods [30 days]. Meanwhile, high doses [800 and 100 ppm] were deleterious to growth and mineralization activity of B. megaterium. The applied malathion up to 400 ppm completely disappeared at the end of the incubation period