Primera evidencia de actividad enzimática nitrilasa en Xylaria sp., y su relación con la biosíntesis de ácido indol-3-acético / First evidence of nitrilase enzymatic activity of Xylaria sp. and its relationship with the biosynthesis of indole-3-acetic acid

Resumen Muchos de los hongos degradadores de madera están implicados en la síntesis de metabolitos bioactivos de naturaleza antimicrobiana y terapéutica, así como de compuestos de importancia biotecnológica, incluyendo derivados indólicos, entre otros. Estos hongos brindan ciertos beneficios ecológicos a las plantas, entre los que se destacan la protección contra fitopatógenos y la promoción del crecimiento radicular. Xylaria sp. es un hongo degradador de celulosa (lignocelulolítico) con potencial biotecnológico. El ácido indol-3-acético (AIA) desempeña un papel sumamente importante en las interacciones planta-microorganismo, ya que es esencial para la fisiología y el correcto desarrollo morfológico vegetal. Se sabe que las enzimas nitrilo-hidrolíticas (nitrilasas) están involucradas en la síntesis de compuestos indólicos en las plantas, no obstante, se dispone de poca información acerca de la naturaleza de estas enzimas en el reino de los hongos. A través de una aproximación bioquímica y de genética molecular, se demuestra por primera vez que Xylaria sp. posee actividad enzimática nitrilasa utilizando compuestos ricos en nitrógeno y carbono como sustrato. La cepa estudiada aumentó sus niveles de expresión génica relativa y mostró crecimiento micelial, ambos en presencia de compuestos químicos como cianobenceno y KCN. Los resultados de este trabajo sugieren que el microorganismo es capaz de degradar moléculas nitrogenadas complejas. Por otra parte, mediante biofertilización con extractos fúngicos, se observó que Xylaria sp. promueve el desarrollo del sistema radicular de plántulas de Arabidopsis thaliana, además de sintetizar AIA.

Abstract Endophytic fungi inhabit plant tissues internally and asymptomatically, and many of them are involved in the synthesis of bioactive metabolites of antifungal and therapeutic nature, as well as other compounds of biotechnological importance including indole derivatives, among many others. Ecologically, they provide some benefits to plants including protection against phy-topathogens and promotion of root growth. In this sense, Xylaria sp. is a cellulose-decomposing fungus with biotechnological potential. It is worth mentioning that indole-3-acetic acid (IAA) also plays an extremely important role in plant-micro-organism interactions, as it is essential for physiology and proper plant morphological development. It is known that nitrile-hydrolytic enzymes (nitrilases) are involved in the synthesis of plant indole compounds; however, relatively little information is available concerning the nature of these enzymes in the fungal kingdom. In view of the above, through a biochemical and molecular-genetic approach, it has been demon-strated for the first time that Xylaria sp. carries out nitrile-hydrolytic enzyme activity using nitrogen and carbonrich compounds as substrate. The studied strain increased its relative gene expression levels and showed mycelial growth, both in the presence of chemical compounds such as cyanobenzene and KCN. Thus, the results of this work suggest that the micro-organism is capable of degrading complex nitrogenous molecules. On the other hand, through fungal biofertilization, it was observed that Xylaria sp. promotes the development of the root system of Arabidopsis thaliana seedlings, in addition to synthesizing IAA.

In vitro bioprospecting of rhizospheric fungi associated to cactus (Opuntia cochenillifera) to plant growth promotion / Bioprospección in vitro de hongos rizosféricos asociados al cactus (Opuntia cochenillifera) para la promoción del crecimiento vegetal

Abstract This study aimed to evaluate in vitro the plant growth promotion capacity of rhizospheric fungi from Opuntia cochenillifera. The isolates were submitted to phosphate solubilization, acetic acid-3-indole (AIA) production, antagonism against phytopathogens, and growth under water activity. All test results were subjected to statistical analysis. The studied isolates were able to solubilize phosphate, being F04 (Aspergillus sp.) and F05 (Coprinellus radians) the ones that presented the highest solubilization indices. As to produce AIA, F02 (Penicillium sp.) and F07 (Aspergillus sp.) were the most promising. All isolates could inhibit the growth of Colletotrichum sp. and Fusarium sp. Water restriction provided mycelial growth for all isolates. Therefore, these characteristics confer characteristics as growth promoters in plants.

Resumen El objetivo de este estudio fue evaluar in vitro la capacidad de los hongos rizosféricos aislados del cactus Opuntia cochenillifera para promover el crecimiento de las plantas. Los aislados fueron sometidos a pruebas de solubilización de fosfato, producción de ácido 3-indol acético (AIA), antagonismo contra fitopatógenos y crecimiento bajo actividad de agua. Todos los resultados de las pruebas se sometieron a análisis estadístico. Los aislados estudiados fueron capaces de solubilizar fosfato, siendo F04 (Aspergillus sp.) y F05 (Coprinellus radians) los que presentaron los mayores índices de solubilización. Sobre la producción de AIA, los que más produjeron fueron F02 (Penicillium sp.) y F07 (Aspergillus sp.). Todos los aislados pudieron inhibir el crecimiento de Colletotrichum sp. e Fusarium sp.. La restricción de agua proporcionó el crecimiento del micelio para todos los aislados. Por lo tanto, estes aspectos les confieren características como promotores del crecimiento en las plantas.

Isolation and Characterization of Plant Growth-Promotion Diazotrophic Endophytic Bacteria Associated to Sugarcane (Saccharum officinarum L.) Grown in Paraíba, Brazil

Abstract: Sugarcane is an important Brazilian commodity, being usually cultivated in soils with low natural fertility. This study aimed to isolate diazotrophic endophytes from sugarcane tissues and evaluate the morphological and physiological characteristics of their colonies as well as their plant growth-promoting (PGP) traits in select diazotrophic endophytic bacteria. Fifty-six bacterial isolates were identified in the sugarcane tissues, and these isolates presented distinct morphological and physiological traits. A total of thirty-five bacterial isolates were biochemically evaluated. Overall, Bacillus was the dominant genus. Isolates of Methylobacterium spp. and Brevibacillus agri were present only in leaves, while Herbaspirillum seropedicae occurred only in stems. Except to IPA-CF45A, all isolates were nitrogenase positive. All endophytes exhibit production of indol 3-acetic acid. Over 50% of endophytes solubilize phosphate, release N-acyl homoserine lactones, and present the activity of 1-aminocyclopropane-1-carboxylic acid deaminase, catalase, lipase and protease. The network analysis showed that isolates belonged to Burkholderia, Herbaspirillum, and Methylobacterium interact with Bacillus. Bacterial endophytes exhibited distinct morphological, physiological, and PGP traits that are useful for sustainable agriculture, highlighting the isolates IPA-CC33, IPA-CF65, IPA-CC9 and IPA-CF27. Further studies on the effects of these diazotrophic endophytes and their potential for providing microbial inoculants for improving sugarcane fields will provide valuable information to maintain the sustainability and environment quality.

IDENTIFICATION OF ENDOPHYTIC BACTERIA OF SEEDS FROM Cedrela odorata L. (Meliaceae) WITH BIOTECHNOLOGICAL CHARACTERISTICS / Identificación de bacterias endófitas de semillas de Cedrela odorata L. (Meliaceae) con características biotecnológicas

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.

Plant growth promoting rhizobacteria and their potential as bioinoculants on Pennisetum clandestinum (Poaceae)

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.

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.

Acute and chronic bovine pulmonary edema and emphysema in Uruguay / Edema e enfisema pulmonar agudo e crônico em bovinos do Uruguai

An outbreak of pulmonary edema and emphysema with acute and chronic cases is reported in a farm in Uruguay. In a herd of 40 Hereford steers, 20 died. The deaths began four days after a change of paddock, from an old pasture of Avena sativa to a lush growing pasture of the same grass. Acutely affected animals showed severe dyspnea, sialorrhea, cough, and subcutaneous edema, and died within 72 hours. Chronically affected steers showed dyspnea, respiratory noises, weight loss, and intolerance to exercise. The deaths began four days after the change of paddock. Ten days after the first death, the steers were withdrawn from the pasture, but continued dying throughout the following 40 days. Twenty animals died and six were necropsied. Grossly, the lungs were diffusely armed and glistening, with reddish and crepitant cut surface, and presented alveolar septae sharply distended by edema and emphysema. There was subpleural emphysema with air blebs distributed across the pleural surface. Presence of Dictyocaulus viviparus was observed in three steers. In some animals, the trachea was diffusely reddish with presence of pink foam; in some others, there was bloody liquid in the tracheal lumen. Histologic examination showed severe diffuse alveolar and interstitial emphysema, hyaline membranes adhered to the alveolar wall, thickening of the interlobular septae with proliferation of type II pneumocytes, and moderate-to-severe multifocal histiocytic, neutrophilic and eosinophilic infiltrate. In the trachea, there was submucosal hemorrhage and moderate multifocal eosinophilic and lymphocytic infiltrate. The steers with chronic signs presented similar lung lesions, but multifocal pulmonary fibrosis and cardiac dilatation were also observed. The diagnosis of acute bovine pulmonary emphysema and edema (ABPE) was based on the occurrence of the disease after introduction of the herd in a lush green pasture, on the characteristic gross and histologic lesions, and on the absence of other toxic or infectious agents causing similar lesions. Cattle raisers should be alert to the risks of occurrence of this disease after the introduction of the herds into paddocks with green and lush pastures.(AU)

Descreve-se um surto de edema e enfisema pulmonar com casos agudos e crônicos em bovinos em uma criação semi-intensiva no Uruguai. De um lote de 40 novilhos da raça Hereford morreram 20. As mortes começaram quatro dias após uma mudança de alimentação, de uma pastagem mais velha de Avena sativa, para uma pastagem recentemente plantada de aveia que estava em brotação. Os animais afetados apresentaram sinais clínicos agudos de dispneia, sialorreia, tosse e alguns desenvolveram edema subcutâneo, morrendo em até 72 horas. Outros novilhos mais cronicamente afetados apresentaram dispneia, ruídos respiratórios, perda de peso e intolerância ao exercício. As mortes começaram quatro dias após a mudança de pastagens. Dez dias após a primeira morte, os novilhos foram retirados do pasto, mas morreram ainda durante 40 dias mais. Ao total, morreram vinte animais e seis foram necropsiados. Nas necropsias dos animais mortos na fase aguda os pulmões estavam difusamente armados e brilhosos e ao corte de coloração avermelhada e crepitante, com os septos alveolares acentuadamente distendidos por edema e enfisema. Havia enfisema subpleural caracterizado por bolhas de ar distribuídas pela superfície pleural. Em três bovinos havia ainda presença de Dictyocaulus viviparus. Alguns animais apresentaram a traqueia difusamente avermelhada com espuma de coloração rósea ou liquido sanguinolento livre na luz traqueal. Histologicamente havia edema e enfisema alveolar e intersticial difuso severo, membranas hialinas espessas aderidas à parede alveolar, espessamento dos septos interlobulares com proliferação de pneumócitos tipo II e infiltrado inflamatório histiocítico, neutrofílico e eosinofílico multifocal moderado a severo. Na traqueia havia hemorragias na submucosa e infiltrado eosinofílico e linfocítico multifocal. Os novilhos com sinais crônicos apresentaram lesões pulmonares semelhantes, entretanto, foram observadas também, fibrose pulmonar multifocal e dilatação cardíaca. O diagnóstico de EEPAB baseou-se na ocorrência da doença após a introdução do rebanho em uma pastagem viçosa em brotação, nas características macroscópicas e histológicas e na ausência de outros agentes tóxicos ou infecciosos que causam lesões semelhantes. Se alerta para os riscos da ocorrência desta enfermidade, quando houver mudanças de pastagens.(AU)

Efecto de ácidos húmicos sobre el crecimiento y la composición bioquímica de Arthrospira platensis / Effect of humic acids on the growth and the biochemical composition of Arthrospira platensis

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.

Rooting of semi-hardwood stem cuttings from current year shoots of Drymis brasiliensis / Enraizamento de estacas caulinares semilenhosas de Drimys brasiliensis oriundas de brotação do ano

ABSTRACT: Drimys brasiliensis Miers is an Angiosperm native to the Atlantic Rainforest, commonly known as cataia. Because of dormancy of its seeds, due to embryonic immaturity, production of cataia seedlings presents challenges regarding propagation of the species. Thus, cuttings emerged as a possible technique to be applied, diminishing plants production time and ensuring uniformity of rooting. Stem cuttings from current year shoots were collected in autumn/2012, prepared with 10-12cm in length, a bevel cut on base and straight on top, keeping two leaves, one leaf or no leaves in the apical portion. After disinfestation, bases of cuttings were submitted to the following treatments with indole-3-butyric acid (IBA) in 50% hydro-alcoholic solution: 100% water, 0, 500, 1500, 3000, 4500 and 6000mg L-1 IBA. A completely randomized experimental design was used, with 3 types of cutting x 7 IBA concentrations, with four replicates of 10 cuttings per experimental unit. After 120 days, the variables percentage of rooted cuttings, number of roots per cutting, length of the three longest roots per cutting, percentage of cuttings with callus, alive and dead, with new shoots and the cuttings maintaining the original leaves were assessed. The application of IBA had no influence on any of the assessed variables. Cuttings with two leaves presented the best rooting percentage (51.1%) and the lowest mortality (5.4%), when compared to cuttings with one leaf (35.0%) or without leaves (0.4%). Cuttings without leaves are to be avoided, since they present the highest mortality percentage (93.6%).

RESUMO: Drimys brasiliensis Miers é uma Angiosperma nativa da Mata Atlântica conhecida popularmente como cataia. Por apresentar dormência em suas sementes devido à imaturidade embrionária, a produção de mudas de cataia apresenta uma problemática a ser desvendada no que se refere à propagação da espécie. Assim, a estaquia foi utilizada como uma possível técnica a ser utilizada, diminuindo o tempo de obtenção das mudas e possibilitando a uniformidade de enraizamento. Estacas caulinares provenientes de brotações do ano foram coletadas no outono/2012, confeccionadas com 10-12cm de comprimento, corte em bisel na base e reto no ápice, mantendo-as com duas, uma e sem folhas na porção apical. Após desinfestação, as bases das estacas foram submetidas aos seguintes tratamentos com ácido indol butírico (IBA) em solução hidroalcoólica 50%: 100% água, 0, 500, 1500, 3000, 4500 e 6000mg L-1 IBA. Foi utilizado um delineamento inteiramente casualizado, com 3 tipos de estacas x 7 concentrações de IBA, com quatro repetições contendo 10 estacas por unidade experimental. Após 120 dias, avaliou-se a porcentagem de estacas enraizadas, número de raízes por estaca, comprimento das três maiores raízes por estaca, porcentagem de estacas com calos, vivas, mortas, com novas brotações e que mantiveram suas folhas originais. A aplicação de IBA não influenciou nenhuma das variáveis estudadas. Estacas com duas folhas apresentaram melhor enraizamento (51,1%) e menor porcentagem de mortalidade (5,4%), quando comparadas com estacas com uma folha (35,0%) ou sem folhas (0,4%). A estaquia sem a presença de folhas não é recomendada, por causar a maior porcentagem de mortalidade (93,6%).

Convergent synthesis of new N -substituted 2-{[5-(1H -indol-3-ylmethyl)-1, 3, 4-oxadiazol-2-yl]sulfanyl}acetamides as suitable therapeutic agents

abstract A series of N-substituted 2-{[5-(1H-indol-3-ylmethyl)-1,3,4-oxadiazol-2-yl]sulfanyl}acetamides (8a-w) was synthesized in three steps. The first step involved the sequential conversion of 2-(1H-indol-3-yl)acetic acid (1) to ester (2) followed by hydrazide (3) formation and finally cyclization in the presence of CS2 and alcoholic KOH yielded 5-(1H-indole-3-yl-methyl)-1,3,4-oxadiazole-2-thiol (4). In the second step, aryl/aralkyl amines (5a-w) were reacted with 2-bromoacetyl bromide (6) in basic medium to yield 2-bromo-N-substituted acetamides (7a-w). In the third step, these electrophiles (7a-w) were reacted with 4 to afford the target compounds (8a-w). Structural elucidation of all the synthesized derivatives was done by 1H-NMR, IR and EI-MS spectral techniques. Moreover, they were screened for antibacterial and hemolytic activity. Enzyme inhibition activity was well supported by molecular docking results, for example, compound 8q exhibited better inhibitory potential against α-glucosidase, while 8g and 8b exhibited comparatively better inhibition against butyrylcholinesterase and lipoxygenase, respectively. Similarly, compounds 8b and 8c showed very good antibacterial activity against Salmonella typhi, which was very close to that of ciprofloxacin, a standard antibiotic used in this study. 8c and 8l also showed very good antibacterial activity against Staphylococcus aureus as well. Almost all compounds showed very slight hemolytic activity, where 8p exhibited the least. Therefore, the molecules synthesized may have utility as suitable therapeutic agents.

resumo Uma série de acetamidas 2-{[5-(1H-indol-3-ilmetil)-1,3,4-oxadiazol-2-il]sulfanila} N-substituídas (8a-w) foi sintetizada em três fases. A primeira etapa envolveu a conversão sequencial de ácido 2-(1H-indol-3-il)acético (1) a éster (2), seguido por hidrazida (3) e, finalmente, a e ciclização na presença de CS2 e KOH alcoólico produziu 5-(1H-indol-3-il- metil)-1,3,4-oxadiazole-2-tiol (4). Na segunda etapa, aminas arílicas/aralquílicas(5a-w) reagiram com brometo de 2-bromoacetila (6​​), em meio básico, para se obter acetamidas 2-bromo-N-substituídas (7a-w). Na terceira etapa, estes eletrófilos (7a- w) reagiram com 4, para se obter os compostos alvo (8a-w). A elucidação estrutural de todos os derivados sintetizados foi realizada por 1H-NMR, IR e técnicas de espectrometria de EI-MS. Além disso, eles foram submetidos a triagem de atividade antibacteriana e hemolítica. Análise da inibição enzimática foi bem apoiada pelos resultados de docking molecular. Por exemplo, o composto 8q exibiu melhor potencial inibitório contra α-glicosidase, e os compostos 8g e 8b exibiram, comparativamente, melhor inibição contra butirilcolinesterase (BChE) elipoxigenase (LOX), respectivamente. Do mesmo modo os compostos 8b e 8c mostraram excelente potencial antibacteriano contra SalmonellaTyphi, semelhante ao do ciprofloxacino, antibiótico padrão usado neste estudo. Os compostos 8c e 8l também mostraram excelente potencial antibacteriano contra Staphylococcus aureus . Quase todos os compostos mostraram pequena atividade hemolítica, sendo que o composto 8p apresentou menor atividade. Assim, as moléculas sintetizadas podem ter a sua utilidade como agentes terapêuticos adequados.

Rooting of semihardwood cuttings of cataia collected in four seasons / Enraizamento de estacas semilenhosas de cataia coletadas em quatro estações

Drymis brasiliensisMiers is a specie of the Winteraceae family, commonly known as cataia or pepper bark. This research was carried out with the aim to study the rooting of semihardwood cuttings, subjected to different concentrations of indolebutyric acid (IBA), collected in four seasons of the year (autumn, winter, spring and summer). Cuttings were prepared with 10-12cm length, keeping two halved leaves. Bases of cuttings were treated as follows: control (100% water), 0; 500; 1500; 3000; 4500 and 6000mg L-1IBA.One hundred and twenty after the planting the following variables were assessed: rooting percentage, number of roots per cutting, length of roots per cutting, cutting callus percentage, cuttings with new shoots and number of cuttings that maintained the initial leaves. The indolebutyric acid is not recommended for rooting of Drimys brasiliensiscuttings. The cuttings collected in winter were the ones that showed greatest rooting percentage (46.96%) for the production of nursery plant of Drimys brasiliensis.

Drimys brasiliensisMiers é uma espécie pertencente à família Winteraceae, popularmente conhecida como cataia. O trabalho foi realizado com o objetivo de avaliar o enraizamento de estacas semilenhosas, submetidas a diferentes concentrações de ácido indol butírico (AIB), coletadas nas quatro estações do ano (outono, inverno, primavera e verão). As estacas foram preparadas com 10-12cm de comprimento e duas folhas cortadas pela metade na porção apical. As bases das estacas foram submetidas aos tratamentos: testemunha (100% água), 0, 500, 1500, 3000, 4500 e 6000mg L-1AIB. Após 120 dias em casa de vegetação, avaliaram-se a porcentagem de estacas enraizadas, o número de raízes por estaca, o comprimento das três maiores raízes por estaca, a porcentagem de estacas com calos, as estacas com novas brotações e as estacas que mantiveram as folhas iniciais. O ácido indolbutírico não é recomendado para o enraizamento de estacas de Drimys brasiliensis.As estacas coletadas no inverno foram as que apresentaram melhor porcentagem de enraizamento (46,96%) para a produção de mudas.

ALLEVIATION OF SALINITY STRESS ON Vicia faba L. PLANTS VIA SEED PRIMING WITH MELATONIN / Disminución del estrés salino en plantas de Vicia faba L. a través de la activación de las semillas con melatonina

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.

Antitumor compounds from Streptomyces sp. KML-2, isolated from Khewra salt mines, Pakistan

BACKGROUND: Actinomycetes are gram positive bacteria with high G + C content in their DNA and are capable of producing variety of secondary metabolites. Many of these metabolites possess different biological activities and have the potential to be developed as therapeutic agents. The aim of the present study was to screen actinomycetes inhabiting halophilic environment such as Khewra salt mines present in Pakistan for cytotoxic and antitumor compounds. RESULTS: An actiomycetes strain designated as Streptomyces sp. KML-2 was isolated from a saline soil of Khewra salt mines, Pakistan. The strain Streptomyces sp. KML-2 showed 84 % cytotoxic activity against larvae of Artemiasalina. In the screening phase, the strain exhibited significant antitumor activity with IC50 values of 12, 48 and 56 µg/ml against Hela, MDBK and Vero cell lines, respectively. After that extract from 20 l fermentation was used to purify secondary metabolites by several chromatographic techniques. Structure elucidation of isolated compounds revealed that it is highly stable producer of Chromomycin SA (1) and 1-(1H-indol-3-yl)-propane-1,2,3-triol (2). Both of the isolated compounds showed significant antitumor activity against Hela and MCF-7 cancer cell lines (IC50 values 8.9 and 7.8 µg/ml against Hela; 12.6 and 0.97 µg/ml against MCF-7, respectively). The 16S rRNA gene sequence (1437 bp) of the strain confirm its identity (99 %) with Streptomyces griseus. CONCLUSIONS: From this research work we were successful in isolating two potent antitumor compounds, Chromomycin SA and 1-(1H-indol-3-yl)-propane-1,2,3-triol from Streptomyces KML-2 strain, isolated from Khewra salt mine. As such this is the second report which confirms that S. griseus can produce Chromomycin SA without introducing any mutagenesis in its biosynthesizing gene cluster and isolated indole derivative is being reported first time from any member of actinomycetes group with having novel antitumor activity against Hela and MCF-7 cells Nucleotide sequences: Nucleotide sequence data reported are available in the GenBank database under the accession number: GenBank KJ009562.

Definição do ponto de colheita comercial e da concentração de etileno exógeno visando melhor aproveitamento do potencial comercial e nutricional de bananas da cultivar Thap Maeo / Definition of harvest time and exogenous etilene concentration aiming at the better commercial and nutritional potential of the bananas of the Thap Maeo cultivar

A banana é um fruto consumido no mundo todo e, ao contrário do que acontece na maioria dos países nos quais predomina o grupo Cavendish, uma grande variedade de cultivares são consumidas nas diferentes regiões do Brasil. No entanto, as informações bioquímicas e fisiológicas a respeito das cultivares consumidas no país são ainda restritas e, apesar de serem muito diferentes entre si, são tratadas da mesma maneira na pré e pós-colheita, o que muitas vezes compromete a qualidade do fruto. Diante disso, dois pontos são fundamentais para a obtenção de frutos de melhor qualidade: a definição do ponto de colheita e o tratamento dos frutos com etileno após a colheita. Hoje a colheita é feita em função do diâmetro dos frutos e o tratamento com etileno é o mesmo para todas as cultivares. Porém, frutos com o mesmo diâmetro nem sempre estão no mesmo estágio de maturidade fisiológica e, usualmente, o tratamento pós-colheita feito com etileno, visando o amadurecimento mais rápido e uniforme dos frutos, não segue nenhuma orientação técnica. A consequência da falta de critérios definidos para a colheita e para a aplicação de etileno resulta em bananas com baixa qualidade e com vida-de-prateleira curta. A produção de banana é dificultada pelos problemas fitossanitários que ocorrem nas plantações, incluindo doenças como as Sigatokas Negra e Amarela e o Mal-do-Panamá. Tendo em vista a ameaça que as doenças da bananeira representam e os prejuízos que podem causar, a introdução de cultivares resistentes é a melhor forma para reduzir a pressão desses patógenos sobre a cultura. Por ser resistente às Sigatokas e ao Mal de Panamá este trabalho visou conhecer melhor a cultivar Thap Maeo (Musa acuminata AAB cv. Thap Maeo) que tem como defeito principal uma vida-de-prateleira muito curta. Os objetivos deste trabalho foram: (1) estabelecer o ponto de colheita das bananas da cultivar Thap Maeo utilizando a metodologia da soma de temperatura a que a planta está exposta durante o desenvolvimento dos frutos; (2) estabelecer, a partir da caracterização físico-química dos frutos, o teor ideal de etileno exógeno para promover o amadurecimento uniforme dos frutos e, (3) estudar o balanço hormonal no amadurecimento dos frutos. Em uma primeira etapa foi implantado um experimento de campo para determinar a Temperatura Base e a Idade Fisiológica Máxima para esta cultivar. Estes parâmetros são necessários para o cálculo da idade fisiológica e na determinação do ponto de colheita. Com o término desta etapa, foram realizadas colheitas de frutos em diferentes épocas do ano para confirmar a metodologia usada. Foi possível estabelecer uma metodologia para estimar o ponto de colheita dos frutos de acordo com a estação do ano em que estes se desenvolveram. Para estabelecer o teor ideal de etileno exógeno aplicado na pós-colheita, os frutos foram testados com diferentes concentrações de etileno (0 a 1000 ppm) em dose única. Os resultados indicam que mesmo as menores concentrações de etileno aplicadas promovem o amadurecimento uniforme dos frutos, sem diferenças aparentes entre os tratamentos. Além disso, as análises do perfil de compostos voláteis da polpa indicam que a maior diferença entre o aroma dos tratamentos é entre frutos tratados e não tratados. Os frutos que não foram tratados apresentaram maior quantidade de compostos identificados. Em todas as colheitas realizadas, o período entre o início do amadurecimento e o amadurecimento completo dos frutos foi consideravelmente curto se comparado a bananas do subgrupo Nanica. Para melhor compreender estes resultados, além do etileno, foram quantificados os teores livres dos hormônios ácido indol-3-acético (AIA) e ácido abcísico (ABA). Em comparação com estudos anteriores, com cultivares do subgrupo Nanica, a cultivar Thap Maeo produz mais etileno e ABA durante o amadurecimento, além de apresentar menores teores de AIA quando verde. Estas diferenças hormonais podem explicar o curto período de amadurecimento desta cultivar

Banana is an important staple food. Unlike the world Market, which has only the Cavendish Bananas as export bananas, Brazilian market has many Bananas varieties. However, biochemical and physiological information about these varieties are still scarce and different bananas cultivars are treated the same way in pre or post-harvest, which often affects the quality of the fruit. In this context, two issues are important to reach best fruit quality: definition of harvest time and the exogenous ethylene treatment. Now a day the harvest time is defined by the diameter of the fruits and the ethylene treatment is equal for all cultivars. However, fruits with the same diameter not always are in the same development grade and the post-harvest ethylene treatment, aiming a faster and uniform ripening, does not follow any technical recommendation. The consequences for the lack of criteria in the harvest and for the ethylene treatment might be a low quality and shelf life banana. Banana production is hampered by plant health problems occurring in the production fields, including the diseases such as the Sigatokas and the Panama Wilt. Given the threat of the banana diseases, and the damage they can cause, the introduction of resistant cultivars is the best way to reduce the pressure of these pathogens on this crop. Being resistant to Sigatokas and Panama diseases, this work aimed to know better the cultivar Thap Maeo (Musa acuminata AAB cv. Thap Maeo) whose main defect is a short shelf life. The objectives of this work were: (1) to establish the harvest time of Thap Maeo bananas using the thermal sum techniques, (2) using the physico-chemical characterization of the fruit, establish the ideal concentration of exogenous ethylene to promote uniform ripening of fruits and (3) study the hormonal balance in the fruit ripening. The first step was a field experiment to determine the base temperature and the maximum physiological age. These parameters are used to calculate the thermal sum. The next step was to harvest fruits from different times of the year to confirm the methodology. It was established a methodology to estimate the harvest time according to the season in which the fruit has developed. Five ethylene concentration were tested (0-1000 ppm). Results showed that even the low ethylene concentration applied could promote the uniform ripening of the fruits with no apparent differences between the treatments. Furthermore, the analysis of volatile compounds in the pulp indicate that the major difference between the aroma of treatments is between treated and untreated fruits. The fruits that were not treated showed a higher amount of identified compounds. For all harvested fruits, the period between the ripening start and full ripening of the fruit was short when compared to the Nanica bananas. Levels of the hormones indole-3-acetic acid and abscisic acid, in addition to ethylene, were quantified to better understand these results. Compared to previous studies, with cultivars of Nanica subgroup, the cultivar Thap Maeo produce more ethylene and ABA during ripening, and have lower IAA level in the green stage. These hormonal differences may explain the short maturity period of this cultivar

Synthesis and DPPH scavenging assay of reserpine analogues, computational studies and in silico docking studies in AChE and BChE responsible for Alzheimer's disease

Alzheimer's disease (AD) is a fast growing neurodegenerative disorder of the central nervous system and anti-oxidants can be used to help suppress the oxidative stress caused by the free radicals that are responsible for AD. A series of selected synthetic indole derivatives were biologically evaluated to identify potent new antioxidants. Most of the evaluated compounds showed significant to modest antioxidant properties (IC50 value 399.07 140.0±50 µM). Density Functional Theory (DFT) studies were carried out on the compounds and their corresponding free radicals. Differences in the energy of the parent compounds and their corresponding free radicals provided a good justification for the trend found in their IC50 values. In silico, docking of compounds into the proteins acetylcholinesterase (AChE) and butyrylcholinesterase (BChE), which are well known for contributing in AD disease, was also performed to predict anti-AD potential.

A doença de Alzheimer (DA) é uma doença neurodegenerativado sistema nervoso central, em rápido crescimento, e antioxidantes ajudam a suprimir o estresse oxidativo causado por radicais livres, responsávies pela DA. Avaliou-se, biologicamente, série de derivados sintéticos de indol selecionados para identificar novos antioxidantes. A maioria dos compostos avaliados apresentou de significativa a boa propriedade antioxidante (valor de IC50 399,07140.0 ± 50 µM). Eftuaram-se estudos de Teoria do Funcional de Densidade (DFT) com os compostos e os seus correspondentes radicais livres. As diferenças de energia entre os compostos protótipos e os radicais livres correspondentes proporcionaram boa justificativa para a tendência encontrada nos seus valores de IC50. O ancoramento in silico dos compostos com a acetilcolinesterase (AChE) e com a butirilcolinesterase (BChE), que contribuem para a DA, foi, também, realizado para prever o seu potencial anti-DA.

Indole butyric acid and substrates influence on multiplication of blackberry 'Xavante' / Influência do ácido indol butírico e substratos na propagação de amoreira-preta 'Xavante'

Blackberry is a shrubby plant specie which has a high economic importance among agriculture crops. Brazil is the major country of Latin America with the highest future scope for blackberries. For availability of good quality and maximum quantity of seedlings, the present study was carried out at the Universidade Estadual de Londrina,PR from January to March in 2013. The aim of the study was to evaluate the multiplication of blackberry 'Xavante' cuttings under different type of substrates treated with different levels of indole butyric acid (IBA). The experiment was laid out in randomized complete block design with 2 factors, i.e., substrate (rice husk, vermiculite and coconut fiber) and IBA (0; 1,000; 2,000 and 3,000mg L-1), with 5 replications. Each replicate consisted of 10 cuttings. The variables studied were: cutting rooting, cutting survival, leaf retention, cuttings with new leaves, number of major roots, length of major roots and roots dry weight. Most of the variables were significantly affected by both substrate and IBA. Rice husk and vermiculite performed better than coconut fiber and provided the same results for most of the variables, while coconut fiber showed lower performance for all of the variables studied. IBA significantly affected the rooting and the number of major roots. It is concluded that for multiplication of blackberry 'Xavante', both rice husk and vermiculite can be used along 2,000mg L-1 of IBA.

A amoreira-preta é uma espécie vegetal arbustiva que apresenta relevante importância econômica entre as culturas frutíferas. O Brasil é o maior produtor da América Latina. Para disponibilidade de boa qualidade e alta quantidade de mudas, o presente estudo foi realizado na Universidade Estadual de Londrina, PR, de janeiro a março, em 2013.O objetivo foi avaliar a mulitiplicação de amoreira-preta 'Xavante' em vários tipos de substratos sob diferentes níveis de ácido indol butírico (AIB). O delineamento experimental foi em blocos casualizados com dois fatores: substrato (casca de arroz carbonizada, vermiculita e fibra de coco) e AIB (0, 1.000, 2.000 e 3.000mg L-1), com cinco repetições, sendo cada repetição constituída por 10 estacas. As variáveis ​​avaliadas foram: estacas enraizadas, sobrevivência das estacas, retenção de folhas, estacas com folhas novas, número de raízes, comprimento de raízes e massa seca dos raízes. A maioria das variáveis foi significativamente afetada pelos substratos e AIB. A casca de arroz e vermiculita apresentaram melhor desempenho quanto à maioria das variáveis, enquanto a fibra de coco apresentou dempenho inferior. O AIB afetou significativamente a porcentagem de enraizamento e o número de principais raízes. Conclui-se que, para a otimizar a multiplicação de mudas de amoreira-preta 'Xavante' , tanto a casca de arroz carbonizada como a vermiculita podem ser usadas, associadas a 2.000mg L-1 de AIB.

The morphology and bioactivity of the rice field cyanobacterium Leptolyngbya / Morfología y bioactividad de la cianobacteria Leptolyngbya en los campos de cultivo de arroz

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.

Exploring the Synthesis of New 1-(4-Substitutedphenylamino) imidazo[1,5-a]indol-3-one Derivatives as Cyclized Analogs of Leucettines.

New 1-arylaminoimidazo[1,5-a]indol-3-ones were synthesized as cyclized derivatives of leucettine L41, a low molecular weight inhibitor of the DYRKs/CLKs protein kinases with potential applications in Alzheimer's disease and Down syndrome. In this first approach, access to the desired 1-aminoimidazo[1,5-a]indol-3-ones involved 5 steps and was explored with a series of various primary amines and polar secondary amines in order to introduce molecular diversity on N-1 position. The 5 step synthesis of the 1-arylaminoimidazo[1,5-a]indol-3-ones was achieved and the limiting step of this process was the final cyclization via a sulphur/nitrogen displacement from methylsulfanyl thiourea intermediates. Good results were obtained for isothioureas derived from primary amines. The 1-arylaminoimidazo[1,5-a]indol-3-ones were evaluated on a panel of five protein kinases (DYRK1A, CK1, CDK5/p25, GSK3 and CLK1).

Influência da salinidade sobre o crescimento e a produção de ácido indol acético de Burkholderia spp. endofíticas de cana-de-açúcar / Salinity influence on the growth and production of indole acetic acid by endophytic Burkholderia spp. from sugarcane

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.

Caracterização e influência de rizóbios isolados de alfafa na germinação e desenvolvimento inicial de plântulas de arroz / Characterization and influence of alfafa rhizobia on germination and early growth of rice seedling

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.

Simple preparation of new N-aryl-N-(3-indolmethyl) acetamides and their spectroscopic analysis / Preparación sencilla de nuevas N-aril-N-(3-indolmetil) acetamidas y su análisis espectroscópico / Preparação simples de novas N-aril-N-(3-indolmetil) acetamidas e sua análise espectroscópica

Objectives. To prepare new indolic molecules and characterize them by spectroscopic methods. Materials and methods: All reagents were purchased from Aldrich, commercial grade. The purity of the products and the composition of the reaction mixtures were monitored by thin layer chromatography over Silufol UV254 0.25 mm-thick chromatoplates. Product isolation and purification were performed by column chromatography (SiO2), using ethyl acetate-petroleum ether mixtures as eluents. Results. The synthesis of new N-aryl-N-(3-indolmethyl) acetamides based on first step iminization reaction of indol-3-carbaldehyde is accomplished. The structures of the C-3 substituted indoles were confirmed by ¹H-NMR and 13C-NMR studies supported by inverse-detected 2D NMR experiments and also through monocrystal X-ray diffraction. Conclusions. An efficient, economic, and fast synthetic route was designed to the construction of the N-aryl-N-(3-indolmethyl) acetamides, structural analogues of some alkaloids.

Objetivos: Preparar nuevas moléculas indólicas y caracterizarlas por los métodos espectroscópicos. Materiales y métodos. Todos los reactivos fueron adquiridos de Aldrich, de grado comercial. La pureza de los productos y composición de las mezclas de reacción fueron monitoreadas por la cromatografía en capa delgada, Silufol UV254 0.25 mm-grosor de cromatoplacas. El aislamiento y purificación de los productos fueron realizados por la cromatografía en columna (SiO2), usando mezclas de acetato de etilo y éter de petróleo como eluentes. Resultados. Se ha realizado la síntesis de nuevas N-aril-N-(3-indolmetil) acetamidas, basada en la reacción de iminización del indol-3-carbaldehído. Las estructuras de los indoles C-3 sustituidos fueron confirmadas por estudios de ¹H, 13C -RMN, experimentos de 2D RMN y también por difracción monocristal de Rayos X. Conclusiones. Se ha diseñado una nueva ruta de síntesis eficiente, económica y rápida para la construcción de las N-aril-N-(3-indolmetil) acetamidas, análogos estructurales de diversos alcaloides.

Objetivos. Preparar novas moléculas indólicas e caracterizar-las através de métodos espectroscópicos. Materiais e métodos. Todos os reagentes foram obtidos de Aldrich, de tipo comercial. A pureza dos produtos e a composição das misturas de reação foram monitoradas por cromatografia em camada fina, Silufol UV254 0,25 mm de espessura das cromatoplacas. O isolamento e a purificação dos produtos foram feitos através de cromatografia em coluna (SiO2), utilizando misturas de acetato de etila e éter de petróleo como eluente. Resultados. Realizou-se a síntese de novas N-aril-N-(3-indolmetil)acetamidas, baseada na reação de iminización do indol-3-carbaldeído. As estruturas dos indóis C-3 substituídos foram confirmadas por estudos de ¹H, 13C -RMN, experimentos de 2D RMN e também por difração monocristal dos Rayos X. Conclusões. Desenhou-se una nova rota de síntese eficiente, econômica e rápida para a construção das N-aril-N-(3-indolmetil) acetamidas, análogos estruturais de vários alcalóides.