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C:\Users\Vaio\Dropbox\GISSELLE-PEDRO\BECA CONICET\TESIS\Tesis Versión Final CD Apud Gisselle 2018\1-Carátulas, Agradecimientos UNIVERSIDAD NACIONAL DE TUCUMAN FACULTAD DE BIOQUIMICA QUIMICA Y FARMACIA Ayacucho 471 - T. E. 0054 381 4107215 - FAX 0054 381 4248169 T4000CAN – San Miguel de Tucumán – República Argentina Trabajo de Tesis Doctoral METABOLITOS SECUNDARIOS DE PLANTAS AUTÓCTONAS DEL NOA: IDENTIFICACIÓN Y CARACTERIZACIÓN DE MOLÉCULAS ÚTILES EN EL CONTROL DE HONGOS TOXIGÉNICOS CAUSANTES DE PODREDUMBRES EN UVA Y CEREALES Licenciada en Biotecnología GISSELLE RAQUEL APUD -2018- UNIVERSIDAD NACIONAL DE TUCUMAN FACULTAD DE BIOQUIMICA QUIMICA Y FARMACIA Ayacucho 471 - T. E. 0054 381 4107215 - FAX 0054 381 4248169 T4000CAN – San Miguel de Tucumán – República Argentina AUTORIDADES HONORABLE CONSEJO DIRECTIVO Mag. Adriana Correa Zeballos Dr. Manuel Javier Aybar Dra. Viviana Andrea Rapisarda Bioq. Esp. Ana Verónica Oldano Dra. Ana Lucrecia Iruzubieta Villagra Dra. María Antonieta Gordillo Bioq. Esp. Vanesa Estela Quiroga Sr. Mario Rodríguez Sr. Joaquín Hernán Vargas Srta. Elizabeth Abigail Gutiérrez Srta. Karen Nahir Ríos DECANO Dr. Edgardo Hugo Cutin VICE-DECANA Dra. Inés del Carmen Ramos SECRETARIA DE ASUNTOS ACADEMICOS Dra. Marta Elena Cecilia JEFA DEL DEPARTAMENTO POSGRADO Lic. Marta Inés Quinteros UNIVERSIDAD NACIONAL DE TUCUMAN FACULTAD DE BIOQUIMICA QUIMICA Y FARMACIA Ayacucho 471 - T. E. 0054 381 4107215 - FAX 0054 381 4248169 T4000CAN – San Miguel de Tucumán – República Argentina AUTORIDADES DEPARTAMENTO DE POSGRADO DIRECTOR: Dr. Sergio Enrique Pasteris CONSEJO TITULAR: Dra. Inés del Carmen Ramos Dra. María Carolina Navarro Dra. Maria Cristina Gaudioso Dra. Paula Andrea Vincent Dra. Maria Cristina Rubio Suplentes Mag. Maria Graciela Benzal Dra. Clara del Valle Silvia de Ruiz Dra. María Inés Nieva Moreno Dra. Claudia Alejandra Crespo Dra. María Angélica Véliz REPRESENTANTE DE POSGRADO ANTE LA SECRETARÍA DE POSGRADO DE LA UNT Dra. Paula Andrea Vincent UNIVERSIDAD NACIONAL DE TUCUMAN FACULTAD DE BIOQUIMICA QUIMICA Y FARMACIA Ayacucho 471 - T. E. 0054 381 4107215 - FAX 0054 381 4248169 T4000CAN – San Miguel de Tucumán – República Argentina TRABAJO DE POSGRADO PARA LA OBTENCIÓN DEL GRADO ACADÉMICO SUPERIOR DE DOCTORA EN BIOQUIMICA CARRERA DE DOCTORADO EN BIOQUIMICA Acreditado y Categorizado A ante la Comisión Nacional de Acreditación Universitaria (CONEAU) Resolución nº: 732/00 Acreditado y Categorizado A ante la Comisión Nacional de Acreditación Universitaria (CONEAU) Resolución nº: 489-CONEAU-12 DIRECTOR: Dr. Manuel Javier Aybar COMITÉ ACADÉMICO: Dra. Aida Ben Altabef Dra. Gladis Susana Álvarez Dra. Inés del Carmen Ramos Dra. Roxana Beatriz Medina Dr. Raúl Armando Salomón UNIVERSIDAD NACIONAL DE TUCUMAN FACULTAD DE BIOQUIMICA QUIMICA Y FARMACIA Ayacucho 471 - T. E. 0054 381 4107215 - FAX 0054 381 4248169 T4000CAN – San Miguel de Tucumán – República Argentina TRABAJO DE POSGRADO TITULADO: METABOLITOS SECUNDARIOS DE PLANTAS AUTÓCTONAS DEL NOA: IDENTIFICACIÓN Y CARACTERIZACIÓN DE MOLÉCULAS ÚTILES EN EL CONTROL DE HONGOS TOXIGÉNICOS CAUSANTES DE PODREDUMBRES EN UVA Y CEREALES TESISTA: Lic. en Biotecnología GISSELLE RAQUEL APUD DIRECTOR: Dr. DIEGO ALEJANDRO SAMPIETRO CODIRECTOR: Dr. PEDRO ADRIÁN AREDES FERNÁNDEZ COMISION DE SUPERVISION: Dra. NANCY ROXANA VERA Dra. LUCÍA MARGARITA MENDOZA A mi familia Agradecimientos A mi Director, Dr. Diego Sampietro por sus conocimientos, sus aportes y su dedicación. Gracias por estar pendiente de mí siempre para que pueda cumplir mi objetivo. A mi Co-director, Dr. Pedro Aredes Fernández por acompañarme desde mis inicios y enseñarme a valorar mi esfuerzo, creer en mí y continuar a pesar de las dificultades. Gracias Pedro por preocuparte siempre, por no dejarme bajar los brazos, por escucharme, por confiar en mí. Me siento orgullosa de poder cumplir otra meta a tu lado. A la Dra. Marta Elena Farías porque aún tengo presente la confianza que depositó en mí para formar parte de su grupo de trabajo. Gracias por transmitirme tus valores y tus conocimientos. Te extraño! Un gran beso al cielo. A la Dra. Nancy Vera y Dra. Lucía Mendoza, miembros de la comisión de supervisión, por brindarme sus conocimientos y los aportes necesarios para que pueda desarrollar esta Tesis. Gracias por su bondad y su apoyo constante. Al Dr. César Catalán por enseñarme y acompañarme en diferentes etapas experimentales de mi Tesis brindándome sus aportes y grandes conocimientos. Al personal del Instituto de Biotecnología, Dr. Roberto Navarro, Dra. Cristina Rubio, Dra. Cristina Maldonado, Dra. Zonnia López y Dra. María Ester Romero por permitirme realizar mi Tesis en esta cátedra, por su amabilidad y su colaboración. A los no docentes Carlos Catania y Gustavo Álvarez por su alegría, amabilidad y respeto. A la Dra. María Antonieta Gordillo por su excelente predisposición, cariño y amabilidad. Gracias por resolver mis dudas y ayudarme siempre que necesité. A Oscar Parravicini, Fernando Díaz y Luis Argañaraz de la Cátedra de Química Orgánica II, por su colaboración, su buena predisposición para ayudarme y su gran amabilidad. Al Departamento de Posgrado de la Facultad de Bioquímica, Química y Farmacia, principalmente a la Lic. Marta Quinteros y Carlos Puig por su gran predisposición para despejar mis dudas y resolver mis inconvenientes. A la Dra. María Gilda Stivala, mi amiga y compañera incondicional, por enseñarme y transmitirme su experiencia, por preocuparse siempre por mí, por entenderme, por su ayuda desinteresada, por sus consejos y palabras de aliento, por su confianza y por los hermosos momentos compartidos. Gracias Gil por estar a mi lado siempre! Sos una excelente persona y profesional y le agradezco a Dios haberte conocido. A la Lic. Cecilia Ledesma por ayudarme siempre y darme alegría con todas sus ocurrencias, por las charlas y los momentos compartidos, por transmitirme su fortaleza. Gracias Ce por ser parte de mi vida y acompañarme en esta etapa. Al Lic. José Martínez Chamás por brindarme su alegría con su risa incomparable, por creer siempre que lo lograría y por su ayuda constante. A la Lic. Irina Kristof por las charlas y mates compartidos, por preocuparse, por destacar mis virtudes, por darme valor y creer en mí. A Melina Belizán, Pamela Terán Baptista, Analía Gómez, Laura Andina y Marisol Jiménez de la Cátedra de Fitoquímica por ser excelentes personas, por su ayuda, colaboración y amabilidad de siempre. A la Dra. Melina Sgariglia y Dr. Rodolfo Soberón por su ayuda, sus aportes y sugerencias. Gracias por la amabilidad y el respeto de siempre. A Baltasar, mi amor, mi amigo, mi compañero, mi confidente, porque me acompañó en toda esta etapa, desde el inicio, y nada hubiera sido igual si no hubiera estado a mi lado. Gracias por creer en mí, por alentarme, por tu infinita paciencia, por tu inmenso amor y tu compañía inigualable…Te amo!! A mi mamá, Raquel, la persona que más me conoce en el mundo, mi amiga fiel y mi compañera incondicional. No me alcanzan las palabras para agradecerle porque cada paso que di estuvo acompañado de sus consejos, sus enseñanzas y su fe en mí. Gracias por incentivarme cada día y enseñarme que todo es posible y se puede lograr. A mi papá, Félix, porque a pesar de la distancia me acompaña, me guía y me alienta en cada paso que doy. Gracias por conocerme, por entenderme, por hablarme con las palabras justas cuando más lo necesito y sobre todo, gracias por demostrar desde siempre sentirte orgulloso de mí. A mi hermano Fede, por su apoyo incondicional y su cariño sincero. Gracias por enseñarme a ser fuerte y enfrentar mis obstáculos, por conocerme y ser mi sostén, por cuidarme, valorarme y aconsejarme. A mi hermana Raquelita por ser mi amiga y confidente, mi cable a tierra, por festejar mis logros, por estar a mi lado siempre, por quererme y creer en mí. Gracias por tu compañía incondicional en esta etapa tan importante para mí. A mi abuela Emilia porque siempre confío en mi y se sintió orgullosa de todos mis logros. Desearía poder tenerte cerca y compartir mi alegría con vos. Un gran abrazo al cielo Sete querida! A mis queridas amigas María Belén Arce y Priscilla Barón, por todos los hermosos momentos compartidos, por acompañarme y estar presente en todo momento de forma incondicional, por incentivarme, escucharme y confiar en mí. A mi amiga Claudia Cequeira, una de las mejores personas que conozco, con la que compartí llantos y alegrías. Gracias por tu fidelidad, tu comprensión y tu cariño inmenso. A mi amiga Celeste Arrieta por su cariño sincero y todos los momentos compartidos. A Reyes, María Celia, Emilia, Julieta, Nicolás y Eugenio por el inmenso cariño que me brindan, por apoyarme siempre y por dejarme formar parte de su familia. ABSTRACT Fungi responsible of rot diseases reduce crops yields and contaminate the harvestable food products with mycotoxins noxious for human and animal health. They include the Fusarium species causing cereal rots, which are fumonisin and trichothecene producers, and Aspergillus species of the Nigri section responsible of black grape rots and ochratoxin A (OTA) production. Chemical control of these fungi is based in the use of commercial fungicides which are expensive, generate environmental pollution and led to the development of fungal resistance. Plant extracts might provide new antifungals overcoming these problems. In this work, the inhibitory effect of extracts from aerial parts (leaves and stems) of native plants from northwest Argentina (Amphilophium cynanchoides, Tecoma stans, Macfadyena cynanchoides and Jacaranda mimosifolia) was investigated on Fusarium (F. graminearum and F. verticillioides) and Aspergillus (A. carbonarius and A. niger) species. Ten of a total of 24 extracts inhibited 50% of the fungal growth (IC50). Microdilution assays indicated that the extracts with the lowest IC50 values were the ethyl acetate leaf and stem extracts of A. cynanchoides against F. graminearum (IC50=1320 ppm) and F. verticillioides (IC50=1305 ppm), respectively, and the dichloromethane stem extract of M. cynanchoides against A. niger (IC50=1025 ppm) and A. carbonarius (1066 ppm). The dichloromethane stem extract of M. cynanchoides was the only one that completely suppressed fungal growth (MIC) at 2500 ppm. It was antifungal on all the fungal strains in dot blot assays where Aspergillus strains were more sensitive than the Fusarium strains.
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