ABSTRACT
ABSTRACT In the present study, 62 endophytic bacterial strains of cedar seeds (Cedrela odorata L.), collected in the municipalities of Huehuetán, Motozintla, and Pijijiapan in the state of Chiapas, Mexico were isolated. The goal was to identify characteristics of biotechnological interest such as biocontrol, promotion ofplant growth, and growth in aromatic compounds. The strains were identified by the partial sequence of the 16S ribosomal gene as belonging to the Bacillus genus. The biocontrol capacity of phytopathogenic fungi, production of indoleacetic acid (IAA), solubilization of phosphate, and growth in xenobiotic compounds (phenanthrene, benzene, anthracene, or phenol) were detected in 26 strains of the 62 isolates. 21 % of the strains inhibited the mycelial growth of Alternaria solani and Fusarium sp., and 13 % of the Phytophthora capsici oomycete. IAA production was detected in 24 isolates, phosphate solubilizing activity was identified in 18 isolates, while the ability to grow in the presence of phenanthrene and benzene was found in 26 isolates; 24 isolates grew in the presence of anthracene and only two isolates grew in phenol as the only carbon sources. This is the first report of the isolation and identification of endophytic bacteria from cedar seeds, where biotechnological characteristics were detected for biological control, promotion of plant growth, and growth in the presence of xenobiotic compounds.
RESUMEN En el presente estudio se aislaron 62 cepas bacterianas endófitas de semillas de cedro (Cedrela odorata L.) colectadas en los municipios de Huehuetán, Motozintla y Pijijiapan en el estado de Chiapas, México, con el objetivo de identificar características de interés biotecnológicas como biocontrol, promoción del crecimiento vegetal y crecimiento en compuestos aromáticos. Las cepas se identificaron por la secuencia parcial del gen 16S ribosomal como pertenecientes al género Bacillus. En 26 cepas de las 62 aisladas se detectaron la capacidad de biocontrol de hongos fitopatógenos, la producción de ácido indolacético (AIA), la solubilización de fosfato y el crecimiento en compuestos xenobióticos (fenantreno, benceno, antraceno o fenol). El 21 % de las cepas inhibió el crecimiento miceliar de Alternaria solani y Fusarium sp., y el 13 % del oomiceto Phytophthora capsici. La producción de ácido indolacético se detectó en 24 aislados y la actividad solubilizadora de fosfato se encontró en 18 aislados, mientras que la capacidad de crecer en presencia de fenantreno y benceno se manifestó en 26 aislados (24 aislados crecieron en presencia de antraceno y solo dos aislados crecieron en fenol como únicas fuentes de carbono). Es importante mencionar que este es el primer reporte del aislamiento e identificación de bacterias endófitas de semillas de cedro, en el que se detectaron características biotecnológicas para el control biológico, la promoción del crecimiento vegetal y el crecimiento en presencia de compuestos xenobióticos.
ABSTRACT
RESUMEN Con el propósito de evaluar el efecto de tres concentraciones de ácidos húmicos (AH) 1, 10, 100 mg/L contra una concentración de ácido indol acético (AIA), sobre la producción de biomasa, pigmentos, proteínas, carbohidratos y lípidos de A. platensis; se realizaron cultivos en discontinuo en medio Zarrouk 25% suplementados con AH y AIA, bajo condiciones de aireación constante y fotoperiodos de 12:12 horas, durante 30 días. Se encontró que la producción máxima de biomasa fue mayor en el cultivo suplementado con 10 mg/L de AH, comparado con el control y los cultivos suplementados con AIA. De la misma manera fue observado en este tratamiento la producción más alta de pigmentos, proteínas y carbohidratos. Por tanto, se logró incrementar el crecimiento y la producción de metabolitos de A. platensis, lo cual permite observar el uso potencial de estas sustancias como estimulantes biológicos orgánicos.
ABSTRACT The purpose of this research was to evaluate the effect of three concentrations of humic acids (HA) 1, 10, 100 mg/L against a concentration of indole acetic acid (IAA) on production of biomass, pigments, proteins, carbohydrates and lipids of A. platensis. Cultures discontinuous in Zarrouk 25% medium supplemented with HA and IAA under conditions of aeration constant and photoperiod from 12:12 hours, during 30 days were made. It was found that the maximum biomass production was higher in the culture supplemented with 10 mg / L of AH, compared to the control and cultures supplemented with AIA. In the same way the highest production of pigments, proteins and carbohydrates were observed in this treatment. Therefore, the growth and production of metabolites of A. platensis was increased, which makes it possible to observe the potential use of these substances as organic biological stimulants.
ABSTRACT
Melatonin is an environmentally friendly-molecule with a potent free radical scavenger and antioxidant capacity. Two pot experiments were conducted during two successive winter seasons (2011/2012 and 2012/2013) at the wire-house of the National Research Centre, Dokki, Cairo, Egypt to study the potentiality of melatonin (100 mM and 500 mM) in alleviating the harmful effect of diluted seawater at a relatively low and high concentrations (3.85 dS/m and 7.69 dS/m, respectively) on the performance of faba bean plants. The results revealed that irrigation of faba bean plants with diluted seawater reduced growth parameters (plant height, leaves number/plant, fresh and dry weights of plant), relative water content (RWC), photosynthetic pigments (chlorophylls a, b and carotenoids), indole acetic acid, total carbohydrate, K+,Ca+2, as well as the ratios of K+/Na+ and Ca+2/Na+. Thiswasaccompanied by significant increases in phenolic content, compatible solutes (total soluble carbohydrate, free amino acids, proline), Na+ and Cl- relative to the control plants (untreated plants). On the other hand, melatonin treatments improved growth parameters, RWC, photosynthetic pigments, total carbohydrate, total phenolic content, indole acetic acid, K+,Ca+2 as well as K+/Na+ and Ca+2/Na+ ratios, either in the plants irrigated with tap water or with diluted seawater, as compared with corresponding controls. Meanwhile, melatonin treatments reduced the levels of compatible solutes, as well as Na+ and Cl- contents, relative to those of corresponding controls. Salinity stress and/ or melatonin treatments induced the production of new protein bands that did not occur in the control plants. Melatonin at 500 mM had a more pronounced effect in alleviating the adverse effects of the two salinity levels under study on the performance of faba bean plants than 100 mM melatonin.
La melatonina es una molécula ambientalmente amigable con una potente capacidad antioxidante y de trampa de radicales libres. Dos experimentos en materas fueron realizados en dos inviernos consecutivos (2011/2012 y 2012/2013) en instalaciones del Centro Nacional de Investigaciones, Dokki, Cairo, Egipto, para estudiar el potencial de la melatonina (100 mM and 500 mM) para disminuir los efectos nocivos del agua de mar diluida a concentraciones relativamente bajas y altas (3,85 dS/m and 7,69 dS/m, respectivamente). Los resultados mostraron que la irrigación de plantas de haba con agua de mar diluida reduce los parámetros de crecimiento (altura de la planta, número de hojas/planta, peso fresco y seco de la planta), el contenido relativo de agua (RWC), los pigmentos fotosintéticos (clorofilas a, b y carotenoides), el ácido indo lacético, los carbohidratos totales, K+, Ca+2, al igual que las relaciones K+/Na+ y Ca2+/Na+. Esto fue acompañado por un incremento significativo en el contenido de fenoles, solutos compatibles (carbohidratos solubles totales, aminoácidos libres, prolina), Na+ y Cl- en comparación con las plantas control (plantas no tratadas). De otro lado, los tratamientos con melatonina mejoraron los parámetros de crecimiento, RWC, los pigmentos fotosintéticos, carbohidratos totales, contenido fenólico total, ácido indo acético, K+,Ca+2 al igual que las relaciones K+/Na+ y Ca+2/Na+ , tanto en las plantas irrigadas con agua dulce de la llave como en las irrigadas con agua de mar diluida en comparación con los controles correspondientes. De otro lado, los tratamientos con melatonina redujeron los niveles de solutos compatibles, al igual que los contenidos de Na+ y Cl-, en comparación con los controles. El estrés por salinidad y/o los tratamientos con melatonina indujeron la producción de nuevas bandas de proteínas que no estuvieron presentes en las plantas control. El tratamiento de melatonina 500 mM tuvo un efecto más pronunciado que el tratamiento de 100 mM en disminuir los efectos adversos de los dos niveles de salinidad estudiados sobre el comportamiento de las plantas de haba.
ABSTRACT
The genus Leptolyngbya comprises filamentous cyanobacteria that are important in rice fields. In the rhizosphere, cyanobacteria produce a variety of secondary metabolites such as auxins that are important in agriculture soil performance. To assess this, Leptolyngbya strain MMG-1, was isolated from the rhizosphere of rice plants and described. For this, the morphology of this strain was studied by light microscopy as well as by confocal laser scanning microscopy. Besides, the ability of this strain to synthesize an auxin-like bioactive compound was demonstrated under various culture conditions (different amounts of tryptophan; pH; different alternating light:dark periods; duration of the incubation). The auxin-like compound was extracted from the culture of Leptolyngbya strain MMG-1 and identified as indole-3-acetic acid (IAA) by thin layer chromatography (TLC) as well as by high performance liquid chromatography (HPLC). Our results showed that the strain required the precursor L-tryptophan for the synthesis of IAA. Leptolyngbya strain MMG-1 accumulated IAA intracellularly. The IAA secreted by Leptolyngbya strain MMG-1 was significantly correlated with the initial concentration of L-tryptophan in the medium, as well as with the duration of the incubation. The bioactivity of the secreted IAA was determined by its effect on the rooting pattern of Pisum sativum seedlings. The culture supernatant of Leptolyngbya strain MMG-1 stimulated the seedling lateral rooting, while it decreased root length. Hence, rhizospheric Leptolyngbya produced auxin under different conditions and affected the plants rooting pattern. Rev. Biol. Trop. 62 (3): 1251-1260. Epub 2014 September 01.
El género Leptolyngbya comprende cianobacterias filamentosas que son importantes en los campos de cultivo de arroz. En la rizosfera, las cianobacterias producen una variedad de metabolitos secundarios, tales como auxinas, que son importantes en el rendimiento de la agricultura del suelo. La cepa Leptolyngbya MMG-1, fue aislada de la rizosfera de plantas de arroz y se describe en este trabajo. La morfología de esta cepa se estudió por microscopía de luz, así como por microscopía confocal de barrido láser. Además, se estimó la capacidad de esta cepa para sintetizar el compuesto bioactivo auxina como se demostró en diversas condiciones de cultivo (diferentes cantidades de triptófano; pH; diferente luz alterna: períodos oscuros; duración de la incubación). La auxina se extrajo a partir del cultivo de la cepa Leptolyngbya MMG-1 y se identificó como ácido indol-3-acético (AA) por cromatografía de capa fina (TLC), así como por cromatografía líquida de alta resolución (HPLC). Nuestros resultados mostraron que la cepa requiere el precursor de L-triptófano para la síntesis de IAA. La cepa Leptolyngbya MMG-1 acumula intracelularmente IAA. El IAA secretada por la cepa Leptolyngbya MMG-1 se correlacionó significativamente con la concentración inicial de L-triptófano en el medio, así como con la duración de la incubación. La bioactividad de la IAA secretada se determinó por su efecto sobre el patrón de enraizamiento de plantas de semillero de Pisum sativum. El sobrenadante del cultivo de la cepa Leptolyngbya MMG-1 estimuló el enraizamiento lateral en la plántula, mientras que se redujo la longitud de la raíz. Por lo tanto, la producción de auxina por Leptolyngbya rizosférica afectó el crecimiento de las plantas.
Subject(s)
Cyanobacteria/physiology , Indoleacetic Acids/isolation & purification , Oryza/microbiology , Cyanobacteria/chemistry , Cyanobacteria/classification , Cyanobacteria/isolation & purificationABSTRACT
A cana-de-açúcar é uma fonte importante de alimento e bioenergia. Assim, o estudo da interação com bactérias endofíticas que fixam o nitrogênio atmosférico (FBN) e produzem o ácido indol acético (AIA) vem sendo amplamente explorado. Porém, devido à sensibilidade bacteriana, tanto a produção de AIA, como a FBN, podem sofrer o impacto de fatores edafoclimáticos. O objetivo deste trabalho foi avaliar a influência de diferentes concentrações de NaCl sobre o crescimento bacteriano, a FBN e a produção de AIA in vitro de linhagens bacterianas do gênero Burkholderia, isoladas endofiticamente de plantas de cana-de-açúcar, cultivadas no estado de Pernambuco. Para tanto, duas linhagens bacterianas endofíticas das espécies Burkholderia gladioli e B. heleia foram inoculadas em meio TSA (com L-triptofano) acrescido de cinco concentrações de NaCl: 0,1; 1; 10; 25 e 50 g.L-1 e incubadas sob agitação. O crescimento bacteriano e a produção de AIA foram avaliados através de um espectrofotômetro por 48 horas. Para o teste de fixação de nitrogênio, as linhagens foram inoculadas em meio LGI-P semi-sólido acrescido de cinco concentrações de NaCl: 0,1; 1; 10; 25 e 50 g.L1 e incubadas a 28ºC por 10 dias. Foi possível observar que altas concentrações de NaCl influenciaram negativamente o crescimento bacteriano e a produção do AIA in vitro por B. glaidoli e B. heleia. E que, concentrações de NaCl acima de 25 g.L-1 inibiram a capacidade de fixar nitrogênio, de ambas as linhagens.
The sugarcane is an important source of food and bioenergy. So, the study of its interaction with endophytic bacteria that fix atmospheric nitrogen (BNF) and produce indole acetic acid (IAA) has been widely explored. However, due to bacterial sensitivity, both the production of IAA as the BNF may suffer the impact of soil and climatic factors. The goal of this study was to evaluate the influence of the NaCl different concentrations on the bacterial growth, on the BNF and on the IAA production in vitro by endophytic bacterial strains of the genus Burkholderia isolated from sugarcane plants grown in the Pernambuco State. In this study, two endophytic bacterial strains of Burkholderia gladioli and B. heleia species were inoculated in TSA (with L-tryptophan) added of five NaCl concentrations: 0.1, 1, 10, 25 and 50 g.L-1 and incubated under agitation. Bacterial growth and IAA production were assessed using a spectrophotometer for 48 hours. The nitrogen fixation test was carried out by the inoculation of the bacteria in semi-solid LGP-I added of five NaCl concentrations: 0.1, 1, 10, 25 and 50 g.L-1 and incubated at 28°C during 10 days. It was observed that high concentrations of NaCl influenced negatively the bacterial growth and indole acetic acid production in vitro by B. glaidoli and B. heleia. Nevertheless, NaCl concentrations above 25 g.L-1 inhibited the ability to fix nitrogen in both strains.
Subject(s)
Sodium Chloride , Saltpetre Soils , Burkholderia , Saccharum , Burkholderia gladioli , Nitrogen Fixation , BiofuelsABSTRACT
A inoculação de plantas leguminosas com rizóbios é um dos principais métodos biotecnológicos de utilização de micro-organismos em plantas visando à fixação biológica de nitrogênio na agricultura. No entanto, nos últimos anos, vêm sendo observada nesses micro-organismos a capacidade de produção de fitohormônios, principalmente o ácido indol-acético (AIA) e a promoção de crescimento em gramíneas. Dessa forma, os objetivos deste trabalho foram quantificar o ácido indol-acético produzido por rizóbios isolados de alfafa, avaliar o efeito da inoculação desses micro-organismos na germinação de sementes de arroz e realizar a caracterização genética desses isolados. Nove rizóbios isolados de nódulos de alfafa foram avaliados quanto a sua capacidade de produção de equivalentes de AIA e a influência da inoculação desses micro-organismos na germinação e desenvolvimento de plântulas de arroz. Os rizóbios produtores de AIA foram identificados pelo sequenciamento da região do gene 16S do DNAr. A produção de equivalentes ao ácido indol-acético foi observada em todos rizóbios, com valores que variaram de 43,04 a 101,26µg mL-1 em meio de cultura. Com relação à germinação das sementes de arroz, a inoculação com rizóbios acelerou o processo e o crescimento de suas plântulas. Os rizóbios UFRGS Ms58, Ms515, Ms195, Ms205, Ms2010 e 2012 foram identificados como pertencentes à espécie Sinorhizobium meliloti e UFRGS Ms55, Ms72 e Ms75 à espécie Rhizobium sp.
The inoculation of leguminous plants with rhizobia is one of the main methods of biotechnological use of microorganisms in order to obtain biological nitrogen fixation in agriculture. However, in recent years it has been attributed to these microorganisms the ability to produce phytohormones, mainly indole acetic acid (IAA), and to promote the growth in grasses. Thus, the objectives of this study were to quantify the indole acetic acid produced by rhizobia from alfalfa and to evaluate the effect of inoculation of these microorganisms on the germination of rice seed and to perform the genetic characterization of these isolates. Nine rhizobia, from nodules of alfalfa, were evaluated for their ability to produce IAA equivalents and for their influence in inoculating these microorganisms on germination and seedling development of rice. Moreover, these rhizobia producers of IAA were identified by the 16S region of DNAr. The equivalent production of indole acetic acid was observed in all tested isolates, with values ranging from 43.04 to 101.26µg mL-1 in culture medium. Regarding the germination of rice seeds, the inoculation with rhizobia accelerated this germination and its growth. Microorganisms UFRGS Ms58, UFRGS Ms515, UFRGS Ms195, UFRGS Ms205, UFRGS Ms2010 and UFRGS 2012 were identified as belonging to the species of Sinorhizobium meliloti. Microorganisms Ms55 UFRGS, UFRGS Ms75 and UFRG Ms72 were identified as belonging to the species of Rhizobium sp.
ABSTRACT
Os rizóbios, conhecidos por sua capacidade de fixar N2 em associação com leguminosas, também se mostram capazes de promover o crescimento de não-leguminosas, especialmente pela produção de ácido indol-acético (AIA). Neste trabalho, objetivou-se selecionar rizóbios produtores de AIA e avaliar o efeito de diferentes concentrações deste fitormônio sobre a germinação e o desenvolvimento inicial de plântulas de alface. Foram selecionados quatro isolados de Bradyrhizobium sp. e um isolado de Rhizobium leguminosarum biovar trifolii, os quais foram crescidos por quatro dias em meio levedura-manitol enriquecido com triptofano. Após esse período, avaliou-se a produção de AIA e procedeu-se à inoculação de sementes de alface com os isolados. O isolado TV-13, de R. leguminosarum biovar trifolii produziu 171,1µg mL-1 de AIA, causando prejuízos para o desenvolvimento das plântulas de alface. Por outro lado, os isolados de Bradyrhizobium sp. produziram entre 1,2 e 3,3µg mL-1 de AIA e aumentaram o vigor das plântulas em relação ao tratamento sem inoculação com rizóbios. Para verificar se essas diferenças foram decorrentes das concentrações de AIA, foram realizados mais dois experimentos, nos quais as sementes foram embebidas em culturas de TV-13 com ou sem a presença de triptofano ou em doses crescentes de AIA sintético. O isolado TV-13 crescido na presença de triptofano causou danos progressivos sobre o desenvolvimento das plântulas de alface, o que não ocorreu na ausência de triptofano. Também foi verificado um retardo na germinação das sementes quando submetidas a altas concentrações de AIA sintético. Os resultados indicam a influência do AIA sobre os parâmetros de germinação, de modo que a inoculação de sementes de alface com rizóbios que produzem baixas quantidades de AIA é uma prática recomendável.
Rhizobia are known by their ability to fix nitrogen in symbiosis with legumes, but they are also capable of promote the growth of non-legume, mainly due to indoleacetic acid production (IAA). In this research, it was aimed to select rhizobia producers of IAA and evaluate the effect of different levels of this hormone over the germination and initial development of lettuce seedlings. Four isolates of Bradyrhizobiumsp. and one isolate of Rhizobium leguminosarum biovar trifolii were grown during four days in yeast manitol medium enriched with tryptophan. After that period, the production of IAA was evaluated and the isolates were inoculated in lettuce seeds. The isolate TV-13, of R. leguminosarum biovar trifolii, produced 171.1µg mL-1 of IAA, causing damages to lettuce seedlings. On other hand, Bradyrhizobium sp. isolates produced between 1.2 and 3.3µg mL-1 of IAA and improved seedlings vigor. In order to verify if these results were due to IAA concentrations, other two assays were carried out, in which lettuce seeds were imbibed in TV-13 cultures with or without tryptophan or in increasing dosages of synthetic IAA. The isolate TV-13 grown in the presence of tryptophan caused progressive damages to lettuce seedlings development, fact that did not occur in the absence of tryptophan. It was also verified a delay in germination of seeds exposed to high levels of synthetic IAA. The results show the influence of IAA on germination parameters, so that the inoculation of lettuce seeds with rhizobia that produce low amounts of IAA is a recommended practice.