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Universidad Austral De Chile Facultad De Ciencias Escuela De Ciencias Universidad Austral de Chile Facultad de Ciencias Escuela de Ciencias PROFESOR PATROCINANTE Alejandra Zúñiga-Feest Instituto de Ciencias Ambientales y Evolutivas Facultad de Ciencias, Universidad Austral de Chile EFECTO DEL FÓSFORO EN EL CRECIMIENTO Y DESARROLLO DE RAÍCES PROTEOIDEAS EN TRES PROTEÁCEAS CHILENAS (Embothrium coccineum, Gevuina avellana y Orites myrtoidea) Tesis de Grado presentada como parte de los requisitos para optar al Grado de Licenciado en Ciencias Biológicas. MARGARITA AURORA DÍAZ SOTO VALDIVIA – CHILE 2012 2 AGRADECIMIENTOS En primer lugar, agradezco a Dios por darme la oportunidad de poder llegar a estas instancias y abrir puertas que veía imposibles, muchas gracias a mis maravillosos padres, Benjamín Díaz C. y Jacqueline Soto S., por todo el apoyo que siempre me han brindado, tanto en lo académico como en lo espiritual. A mi hermano Benjamín Díaz y a mi hermana Silvana Ulloa, por estar en cada momento de mi vida, entregándome su cariño y el ánimo de seguir adelante. La realización de este trabajo, no habría sido posible sin el apoyo y consejo de la Dra. Alejandra Zúñiga, quien me entregó las herramientas necesarias para completar este ciclo, además de su compañerismo y confianza, a la Dra. Susana Valle. Junto a ellas, agradezco el apoyo del equipo del laboratorio de Fisiología Vegetal: Ángela, Andrea, Fernanda y Vanessa. Y a mis compañeras y amigas de la Universidad. No puedo dejar de agradecer también a todos mis amigos, jóvenes y hermanos en la Fe de mi amada Iglesia Pentecostal de Chile que día tras día me demostraban su cariño y oraban por mí. Gracias a mis amados pastores que siempre tenían un consejo sabio en los momentos difíciles. Y por supuesto, a la Escuela de Ciencias, al Instituto de Ciencias ambientales y evolutiva de la Universidad Austral de Chile y al Proyecto Fondecyt Nº 11080162, que financió este trabajo. 3 A mi Dios y mi familia… 4 INDICE DE CONTENIDOS Páginas RESUMEN ...................................................................................................................................... 9 ABSTRACT .................................................................................................................................. 10 1. INTRODUCCIÓN..................................................................................................................... 11 FAMILIA PROTEACEAE, RAÍCES PROTEOIDEAS (RP) Y EXUDACIÓN DE ÁCIDOS ORGÁNICOS ......... 12 1.2 ROL ECOLÓGICO DE LAS RAÍCES PROTEOIDEAS...................................................................... 13 1.3 LAS RAÍCES PROTEOIDEAS SON INDUCIDAS EN CONDICIONES DE DEFICIENCIA DE FÓSFORO... 14 1.4 CONTENIDO DE FÓSFORO, MECANISMO DE ALMACENAMIENTO Y CRECIMIENTO EN VEGETALES .................................................................................................................................................... 15 1.5 TOXICIDAD DEL FÓSFORO EN VEGETALES ............................................................................. 16 1.6 FORMULACIÓN DEL PROBLEMA ............................................................................................. 18 1.7 PROTEÁCEAS CHILENAS ........................................................................................................ 19 1.7.1 Orites myrtoidea ............................................................................................................ 20 1.7.2 Embothrium coccineum. ............................................................................................... 21 1.7.3 Gevuina avellana ........................................................................................................... 22 1.8 HIPÓTESIS.............................................................................................................................. 24 1.9 OBJETIVOS............................................................................................................................. 25 1.9.1 Objetivo general: ........................................................................................................... 25 1.9.2 Objetivos específicos:................................................................................................ 25 2. MATERIALES Y MÉTODOS.................................................................................................. 26 5 2.1 PROCEDENCIA DEL MATERIAL VEGETAL................................................................................ 26 2.2. DISEÑO EXPERIMENTAL........................................................................................................ 26 2.3 MEDICIÓN DE BIOMASA ......................................................................................................... 28 2.4 BIOMASA EN CULTIVO BAJO INVERNADERO .......................................................................... 29 2.5 EVALUACIÓN DE VITALIDAD POR ASPECTO............................................................................ 29 2.6 EVALUACIÓN DEL NÚMERO DE RP......................................................................................... 30 2.7 MORFOLOGÍA DE RAÍCES PROTEOIDEAS................................................................................. 30 2.8 EXUDACIÓN DE RP ................................................................................................................ 31 2.9 ANÁLISIS DE FÓSFORO INTERNO (FOLIAR) ............................................................................. 32 2.10 ANÁLISIS ESTADÍSTICOS ...................................................................................................... 32 3. RESULTADOS ......................................................................................................................... 33 3.1 EFECTO DEL P SOBRE EL CRECIMIENTO EN ALTURA DE LAS PLÁNTULAS. .............................. 33 3.2 EFECTO DEL P EN LA BIOMASA .............................................................................................. 35 3.2.1 Biomasa total ................................................................................................................. 35 3. 3 EFECTO DEL FÓSFORO EN LA FORMACIÓN RP Y RAÍZ TOTAL ................................................ 40 3.4 CONCENTRACIÓN DE FÓSFORO FOLIAR POR TRATAMIENTOS.................................................. 43 3.5 EFECTO DEL P EN LA CALIDAD MORFOLÓGICA Y SOBREVIVENCIA DE LAS PLANTAS.............. 44 3.6 MORFOLOGÍA DE RAÍCES PROTEOIDEAS ................................................................................ 46 3.6.1 Morfología RP a simple vista. ....................................................................................... 46 3.6.2. Morfología de RP electrónica....................................................................................... 48 3.7 EXUDACIÓN ÁCIDA DE RP .................................................................................................... 50 4. DISCUSIÓN.............................................................................................................................. 51 6 4.1 EFECTO DEL FÓSFORO A NIVEL MORFOLÓGICO. ..................................................................... 51 4.2 EFECTO DEL FÓSFORO A NIVEL FOLIAR.................................................................................. 54 4.3 EXUDACIÓN DE ÁCIDOS ORGÁNICOS...................................................................................... 56 5. CONCLUSIONES..................................................................................................................... 57 6. LITERATURA CITADA.......................................................................................................... 58 7. ANEXOS ….............................................................................................................................. 71 INDICE DE FIGURAS Figura 1. Efecto del abastecimiento de P en la velocidad de absorción de fósforo (P) 17 en dos especies con regulación contrastante, bajo condiciones de invernadero: Grevillea crithmifolia y Hakea prostrata Figura 2. Rango de distribución de las especies de Proteácea chilenas 20 Figura 3. Cultivo Plántulas de Embothrium coccineum, Gevuina avellana y Orites 27 myrtoidea Figura 4. Representantes de cada nivel para evaluación de calidad morfológica en 30 plántulas de E. coccineum, G. avellana y O. myrtoidea Figura 5. Exudación de ácidos en clusters de raíces proteoídeas 31 Figura 6. Efecto del fósforo en la altura promedio inicial – final en E. coccineum, G. 34 avellana y O. myrtoidea. Figura 7. Biomasa inicial, seca total de E. coccineum, G. avellana y O. myrtoidea 36 7 Figura 8. Distribución de biomasa inicial de E. coccineum, G. avellana y O. 36 myrtoidea. Figura 9. Distribución de biomasa inicial de E. coccineum, G. avellana y O. 37 myrtoidea Figura 10. Efecto del fósforo en la distribución de biomasa final de E. coccineum, G. 39 avellana y O. myrtoidea. Figura 11. Efecto del fósforo en el Número de RP inicial – final en E. coccineum, G. 41 avellana y O. myrtoidea. Figura 12. Efecto del fósforo en el N° promedio de RP y el peso relativo RP/Raíz total 42 en E. coccineum, G. avellana y O. myrtoidea Figura 13. Imágenes de RP representativas en E. coccineum 46 Figura 14. Imágenes de RP representativas en G. avellana 47 Figura 15. Imágenes de RP representativas en O. myrtoidea 47 Figura 16. Microscopia electrónica de raíces proteoideas de O. myrtoidea 48 Figura 17. Microscopia electrónica de raíz proteoideas completa de O. myrtoidea 49 Figura 18. Detección de la acidificación de la rizófera en placas de agar, con púrpura 50 de bromocresol
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