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Pereira, Susana. 2014 04 21 Tesis Doctoral Aspergillus spp. en plantas de maní nativo y cultivado. Identificación, capacidad toxicogénica y control biológico Pereira, Susana 2014-04-21 Este documento forma parte de la colección de tesis doctorales y de maestría de la Biblioteca Central Dr. Luis Federico Leloir, disponible en digital.bl.fcen.uba.ar. Su utilización debe ser acompañada por la cita bibliográfica con reconocimiento de la fuente. This document is part of the doctoral theses collection of the Central Library Dr. Luis Federico Leloir, available in digital.bl.fcen.uba.ar. It should be used accompanied by the corresponding citation acknowledging the source. Cita tipo APA: Pereira, Susana. (2014-04-21). Aspergillus spp. en plantas de maní nativo y cultivado. Identificación, capacidad toxicogénica y control biológico. Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Cita tipo Chicago: Pereira, Susana. "Aspergillus spp. en plantas de maní nativo y cultivado. Identificación, capacidad toxicogénica y control biológico". Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. 2014-04-21. Dirección: Biblioteca Central Dr. Luis F. Leloir, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires. Contacto: [email protected] Intendente Güiraldes 2160 - C1428EGA - Tel. (++54 +11) 4789-9293 UNIVERSIDAD DE BUENOS AIRES Facultad de Ciencias Exactas y Naturales Departamento de Biodiversidad y Biología Experimental Aspergillus spp. en plantas de maní nativo y cultivado. Identificación, capacidad toxicogénica y control biológico. Tesis presentada para optar al título de Doctor de la Universidad de Buenos Aires en el área CIENCIAS BIOLOGICAS Licenciada Susana Pereira Directoras de tesis: Dra. Silvia Lopez Directorasde tesis:: Dra. Graciela Vaamonde Consejera de Estudios: Dra. Silvia Lopez Lugar de trabajo: Universidad de Buenos Aires, Facultad de Ciencias Exactas y Naturales, Departamento de Biodiversidad y Biología Experimental. PRHIDEB - CONICET. Buenos Aires, 2013 A quienes quiero agradecer: A todos, a tantos… En primer lugar, gracias Daniel, porque me diste un lugar y la oportunidad de trabajar en lo que quiero, cuando ya creía que era imposible. Gracias por confiar en mí. Mis Directoras: Graciela, que me acompañaste desde el primer momento y Silvia que fuiste capaz de “adoptarme” en ese momento tan triste que nos tocó vivir. Memo: Mi primer maestro y quien me ayudó a formar la base. Andre, gracias por “bancarme” todos estos meses con tanto cariño. Ni bien termine, me pongo al día con la Colección… Al fin el “Código de Nomenclatura” nos reconoció la importancia de “mis Aspergillus”… Ceci y Vicky por considerarme una más desde que entré al grupo, por acompañarme, escucharme y aconsejarme siempre. Leo, me enseñaste a aislar endófitos de pastos… ¿Te acordás? y terminamos analizando genes de Aspergillus…¡Casi lo mismo!. Gracias Leo, Ceci, Caro y Esteban, mis salvavidas moleculares!!!!!!. Las chicas del Labo: Su, Caro, Lili, son unas genias, unas amigas, siempre al pie del cañón para ofrecerme ayuda en todo ¿Su, tenés una cajita? ¿Tenés un numerito de BAFC?. Siempre atentas para cerrar todo detrás de mí cuando me voy corriendo (Grande Lili, pero no me apagues la música), para despertarme cuando estoy rendida (en estos últimos tiempos, ojo…). ¿Caro, hoy te toca hacer mate, cierto? Gracias Silvi por estar siempre, menos mal que a veces me ayudás a lavar las cajas, ¡son tantas! Los chicos y chicas de los otros labos: Pato, Vicky “chiquita”, Agus, Ale, Esteban, Esteban “the new”, Fer, Andres, Laurita, Rita, Franco, espero no olvidarme de nadie, los años no vienen solos y somos tantos… Ah!! Ojo que soy la segunda responsable de “La Noche de los Museos” Stella, una ídola, ¡qué días aquellos en que cuantificábamos ocra con mi panza doble! Andrea, Gabi, Pipi, Luli, Virginia, Graciela, Nilda y todas las “chicas” de Virginia que siempre me trataron como si me conocieran desde siempre. Gerardo Cueto, por tu invaluable ayuda al momento de analizar los datos. ¡Cuánto sabés de estadística! Todavía te debo la torta… Al Grupo de pronóstico del Depto. de Ciencias de la Atmósfera y los Océanos que amablemente me cedió los datos climatológicos de Ciudad Universitaria. Mamá, Papá, gracias por estar siempre conmigo y confiar en mi capacidad. Mis hermanos, cuñados y sobrinos, siempre listos para ocuparse de mis nenes por un rato para que yo pueda avanzar...¿Y Susi, todavía te falta mucho?. Gracias Alicia por estar siempre pendiente de nosotros. FEDE, JULI, GONZI, AGUS (TIN TÍN), mis principitos… aún con lo chiquitos que son, fueron capaces de acompañar a una mamá loca que no deja de estudiar ¿Mamá adicta al estudio / trabajo? JOSE, mi compañero y el hombre de mi vida. Si tuviera la oportunidad de volver a vivir, te elegiría otra vez. Gracias por llevar adelante “todo” mientras yo estaba atada a la compu o en la facu hasta tan tarde… y gracias porque a pesar del cansancio te queda un resto para mimarme y alentarme siempre… no podría hacer esto sin vos… A mamá y papá, que me trajeron a este mundo… A Jose, mi compañero en cada paso… A Fede, por tus manos siempre listas para ayudarme… A Gonzi, por tu sonrisa… A Juli, por tus abrazos… A Agus, por tus besos… …Lo que caracteriza al hombre de ciencia no es la posesión del conocimiento o de verdades irrefutables, sino la búsqueda desinteresada e incesante de la verdad… Karl Popper Resumen Aspergillus spp. en plantas de maní nativo y cultivado. Identificación, capacidad toxicogénica y control biológico. Las especies del género Aspergillus son productoras de aflatoxinas, ácido ciclopiazónico y ocratoxina A, importantes por su impacto en la salud humana. El maní es un cultivo económicamente importante en Argentina, susceptible a la colonización por A. flavus y A. parasiticus, con posterior contaminación con aflatoxinas. A fin de reducir dicha contaminación, surgió el biocontrol como alternativa de manejo. Se aislaron Aspergillus spp. a partir de diferentes sustratos de plantas nativas y cultivadas. Se estudió la capacidad toxicogénica en las secciones Nigri y Flavi y la contaminación natural de toxinas en maní, a fin de realizar ensayos de biocontrol in vitro. Se analizó la cinética de crecimiento de cepas seleccionadas como potenciales biocontroladores. Los resultados muestran que las secciones Flavi y Nigri resultan más abundantes en plantas cultivadas, mostrando una elevada habilidad competitiva para colonizar la semilla. La distribución de quimiotipos y esclerocios en A. flavus, podría relacionarse con factores climáticos. Muy bajo porcentaje de cepas de A. niger resultaron ocratoxicogénicas, ninguna asociada a semilla. Su mayor frecuencia en este sustrato podría determinar una menor incidencia de la sección Flavi, debido tal vez a su cinética de crecimiento. Estas características apoyan la hipótesis sobre la capacidad de A. niger como biocontrolador de cepas toxicogénicas de Aspergillus de la sección Flavi. Se encontraron tres cepas atoxicogénicas (A. niger 1636, A. flavus 1639 y A. parasiticus 1640) cuyas características demostraron que podrían ser utilizadas como agentes de control biológico. Palabras clave: Aspergillus – maní – biocontrol – aflatoxinas – ácido ciclopiazónico – ocratoxina A – especies nativas de Arachis– Aspergillus flavus – Aspergillus parasiticus – Aspergillus niger – Abstract Aspergillus spp. in native and cultivated peanut plants. Identification, toxigenic capacity and biological control. The genus Aspergillus contains several species capable of producing aflatoxins, cyclopiazonic acid and ochratoxin A, all important for their impact on human health. The peanut is an economically important crop in Argentina and one of the crops most susceptible to colonization by A. flavus and A. parasiticus from the soil, with the subsequent aflatoxin contamination. In order to reduce the effects of mycotoxins, biological control emerged as an alternative. Aspergillus spp. were isolated from different substrates of native and cultivated plants. Toxigenic capacity in sections Flavi and Nigri and natural contamination in peanuts by toxins were studied, in order to perform biological control tests in vitro. The results show that sections Flavi and Nigri are often found in cultivated and native peanuts, although they are more abundant in cultivated plants, showing a higher competitive ability to colonize the seed. The distribution of chemotypes and sclerotia of A. flavus could be related to climatic factors. A very low percentage of A. niger strains were ochratoxin-producers, and none were associated with seeds. Its higher frequency on this substrate could determine a low incidence of Flavi section, perhaps due to its growth kinetics. These features support the hypothesis that A. niger could be a good biocontrol agent against toxigenic strains of Aspergillus section Nigri. Three atoxigenic strains (A. niger 1636, A. flavus 1639 and A. parasiticus 1640) showed characteristics that suggest that they could be used as biological control agents. Key words: Aspergillus – peanut – biocontrol – aflatoxin – cyclopiazonic acid – ochratoxin A – wild Arachis species – Aspergillus flavus – Aspergillus parasiticus – Aspergillus niger – ÍNDICE GENERAL INTRODUCCIÓN GENERAL 1 1. EL GÉNERO ASPERGILLUS 1 1.1 Importancia del género 1 1.2 Diagnosis 2 1.3 Sistemática 4 1.4 Características morfológicas de importancia para la identificación 6 1.5 Breve descripción de Subgénero Circumdati, sección Flavi 7 1.6 Breve descripción de Subgénero Circumdati, sección Nigri 13 1.7 Micotoxinas 16 1.7.1 Aflatoxinas
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