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El Género Hypocrea Fr. (Hypocreales, Ascomycota) En La Argentina

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El Género Hypocrea Fr. (Hypocreales, Ascomycota) En La Argentina Tesis Doctoral El género Hypocrea Fr. (Hypocreales, Ascomycota) en la Argentina. Estudio de la variabilidad molecular de su estado anamórfico Trichoderma Barrera, Viviana Andrea 2012 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: Barrera, Viviana Andrea. (2012). El género Hypocrea Fr. (Hypocreales, Ascomycota) en la Argentina. Estudio de la variabilidad molecular de su estado anamórfico Trichoderma. Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. Cita tipo Chicago: Barrera, Viviana Andrea. "El género Hypocrea Fr. (Hypocreales, Ascomycota) en la Argentina. Estudio de la variabilidad molecular de su estado anamórfico Trichoderma". Facultad de Ciencias Exactas y Naturales. Universidad de Buenos Aires. 2012. 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 Biologa Experimental —El género Hypocrea Fr. (Hypocreales, Asco ycota) en la Argentina. Estudio de la variabilidad olecular de su estado ana #rfico Trichoder a' Tesis presentada para optar por el t(tulo de Doctor de la Universidad de Buenos Aires en el *rea Biolog(a. Lic. ,iviana Andrea Barrera Directora- Dra. .o ero, Andrea Irene Directora asistente- Dra. 0asoni, A elia Laura Conse1era de estudios- Dra. Lope2, Silvia Edith Buenos Aires, 2012 —El género Hypocrea Fr. (Hypocreales, Ascomycota) en la Argentina. Estudio de la variabilidad molecular de su estado anamrfico Trichoderma En la presente tesis se informan los resultados de un estudio polifsico integral del género Hypocrea/Trichoderma de Argentina. El estudio se encar desde varios puntos de vista: biodiversidad, morfologa, filogenia, incluyendo el anlisis de la variabilidad de cepas biocontroladoras de T. harzianum. Para mostrar los resultados de forma ordenada se a organizado el texto en dos captulos. El Captulo I trata sobre la identificacin de e%emplares coleccionados y depositados en erbarios nacionales e internacionales. En el Captulo II se analiza una coleccin de cepas bicontroladoras nativas de T. harzianum para reconocer caracteres &tiles para su aplicacin en el campo del control biolgico. Como consecuencia de este estudio se identificaron 38 especies de las cuales se incorporan 4 especies nuevas para la ciencia, 2 especies que se citan por primera vez para ,udamérica, 1. especies que se citan por primera vez para la Argentina y por primera vez se cita la relacin teleomorfo-anamorfo para T. longibrachiatum. ,e caracterizaron 20 cepas nativas biocontroladoras de T. harzianum, se observaron diferencias en cuanto a sus requerimientos nutricionales y alta similitud entre sus caracteres genéticos. ,e encontraron 211 marcadores moleculares polimrficos de 2AP3 de los cuales 10 fueron especficos para 0 cepas y 08 marcadores moleculares polimrficos de 4P-PC2 de los cuales 0 resultaron especficos para una cepa. Palabras claves: Hypocrea/Trichoderma, filogenia, tef1, requerimientos nutricionales, marcadores moleculares, 2AP3, 4P-PC2. —Hypocrea Fr. (Hypocreales, Ascomycota) in Argentina. Studies on the molecular variability of its anamorphic state Trichoderma In t is t esis we report t e results of a compre ensive multi-p ase study of t e genus Hypocrea/Trichoderma in Argentina. T e study was faced from several points of view: biodiversity, morp ology and p ylogeny, including analysis of t e variability of biocontrol strains of T. harzianum. To display t e results in an orderly way, t e text is organized into two c apters. C apter I deals wit t e identification of specimens collected and deposited in national and international erbaria. C apter II discusses a collection of native biocontrol T. harzianum strains to recognize c aracters useful for application in t e field of biological control. As a result of t is study 38 species were identified of w ic are incorporated 4 new species to science. Two species are recorded for t e first time for ,out America, 1. species recorded for t e first time for Argentina and for t e first time cited t e teleomorp -anamorp relations ip to T. longibrachiatum. Twenty five native biocontrol T. harzianum strains were c aracterized, s owing differences in t eir nutritional requirements and ig genetic similarity between t eir c aracters. 7e found 211 polymorp ic 2AP3 molecular mar8ers of w ic 10 were specific for 0 strains and 08 polymorp ic molecular mar8ers of 4P-PC2 of w ic 0 were specific for one strain. 9ey words: Hypocrea/Trichoderma, p ylogeny, tef1, nutritional requirements, molecular mar8ers, 2AP3, 4P-PC2. Agradecimientos A la Dra. Andrea Romero por toda su dedicacin y enseanzas, as como su compaerismo y amistad. A la Dra. Laura Gasoni por brindarme todo su apoyo para llevar adelante este doctorado y darme la oportunidad para desarrollar mis proyectos. Al Dr. Leopoldo Iannone por su ayuda desinteresada, por el tiempo dedicados a las consultas y orientacin en filo enias. A la Dra. Carolina Martnez por la ua en el trabajo con los marcadores moleculares, su apoyo, compaerismo, amistad, por estar siempre. A la Prof. Nanci Lpez por estar siempre a mi lado dedicando mucho esfuerzo y ale ra a las lar as horas de trabajo. Al Dr. Die o Sau(a por la buena onda y su enerosidad, por poder contar siempre y la lectura crtica. Al )il. Rodri o Rojo por tantas horas de charlas, debates, intercambio de ideas, compaerismo al extremo. Al Lic. Matas Zapiola por el soporte inform,tico, toda la paciencia y sus a udas observaciones -ue siempre terminan en risas. Al In . Guido Chiessa por toda la ayuda profesional sin olvidarse de las risas -ue nunca faltaron. A la Lic. Mariana Capdet por el ran aporte de colecciones, por compartir horas de trabajo con paciencia y amistad. A la Dra. Adriana .lad(i por esos viajes de coleccin imperdibles por esa bella Tucum,n, los llevar0 siempre conmi o. Al Dr. Ricardo Comerio por toda la buena onda, sabios consejos y compartir el entusiasmo por el microscopio. A la In . )enintende por sus enseanzas sobre el trabajo con bacterias y los buenos chistes. Al In . Rub0n Larossa por su tiempo brindado en la discusin de temas estadsticos. A la Prof. Lorena Lafuente y la Lic. 1stela 2avret por todo el apoyo biblio r,fico y ese lu ar c,lido y tran-uilo en la biblioteca. Al Dr. Roberto Lecuona y el staff del INTA por todo el apoyo brindado desde la institucin. A todos los compaeros de trabajo Carmia, Martita, Pedro, 3uan Carlos, Mnica, Gachy por el buen humor para sobrellevar las lar as horas de trabajo. Al PROPLAM15PR.ID1)5CONIC1T por todo su apoyo y a sus inte rantes por brindar el espacio para el desarrollo acad0mico. A 3ICA por el soporte econmico en el marco del proyecto y la oportunidad de realizar experiencias profesionales en 3apn. A mis hijos 6anina y Tom,s -ue me han comprendido en todo momento, a mis padres, a mis hermanas 2abiana y Romina, 3oa-un, 2abi,n, Co-ui, Cynthia, 2acundo, 1li, toda la familia y ami os por estar siempre y tenerme paciencia. A Daniel -ue me ha brindado cario, apoyo, trabajo, paciencia, fuerza en todo momento. ÈNDICE GENERAL PORTADA TITULO Y RESUMEN EN CASTELLANO TITULO Y RESUMEN EN INGLES AGRADECIMIENTOS ÈNDICE GENERAL CAPITULO I —Estudios taxonmicos del g nero Hypocrea/Trichoderma en Argentina# Pg. INTRODUCCIN 1-10 MATERIALES Y MÉTODOS 11-20 1- Muest as y sitios de muest eo 11 1.1. Colecciones 11 1.2. Muest as de suelo 12 1.3. Muest as de a+ces 13 1.,. Espec+menes estudiados de .e ba io. 13 1.0. Obse 1aciones mic osc2picas 13 2- Aislamiento de las cepas 1, 2.1. 3uentes de las cepas 1, 2.2. Conse 1aci2n de los aislamientos 14 2.3 Tasa de c ecimiento 17 3- Estudio 3ilogen6tico 17 3.1. E7t acci2n de ADN 17 3.2. Cuantificaci2n del ADN e7t a+do 18 3.3. Ma cado es Gen6ticos 18 3.,. Elect ofo esis pa a e1aluaci2n de p oductos de amplificaci2n 19 3.0 Secuenciaci2n 19 3.4. Anlisis filogen6tico 19 ,. O denamiento y modalidad de las desc ipciones 20 RESULTADOS 21-80 1. Colecciones 21 2. Aislamiento de las cepas 21 3. Especies identificadas 21 ,. Desc ipciones y discusi2n de las especies 22-80 Secci2n Longib ac.iatum 22 H. longibrachiata / T. longibrachiatum 22 H. nothoandinensis 2, H. schweinitzii / T. citrinoviride 24 T. ghanense 29 T. parareesei 30 T. saturnisporum 31 Secci2n T ic.ode ma Clado Rufa 33 H. atroviridis /T. atroviride 33 T. gamsii 3, T. aff. koningii 34 H. koningiopsis / T. koningiopsis 37 Trichoderma sp. no1. 1 39 T. ovalisporum ,1 T. aff. stilbohypoxyli ,3 Hypocrea sp. 1 ,, Trichoderma sp. no1. 3 ,4 Clado Pac.ybasium A ,7 T. asperellum ,7 T. hamatum ,8 Trichoderma sp. no1. 2 ,9 Secci2n Pac.ybasium B 00 Clado C.lo ospo a 00 H. cremea / 00 T. cremeum 01 H. sinuosa 02 T. sinuosum 03 Clado Li7ii / Catopt on 0, H. catoptron / T. catoptron 0, H. lixii / T. harzianum 00 Clado Vi ens 09 H. virens / T. virens 09 Clado Ce amica 40 H. ceramica 40 Clado Lutea 41 H. melanomagna / T. melanomagnum 41 >Lone Lineages? 42 H. eucorticioides 42 H.
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