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Bioprospección Química Y Propagación De Plantas Nativas Del Monte Patagónico Como Estrategias De Conservación Y Uso Sust

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Bioprospección Química Y Propagación De Plantas Nativas Del Monte Patagónico Como Estrategias De Conservación Y Uso Sust “Bioprospección química y propagación de plantas nativas del monte patagónico como estrategias de conservación y uso sustentable” TESIS DOCTORAL Autor: Patricia Alejandra Boeri Director: Dra Sandra Sharry Codirectores: Dr Daniel Barrio Dra Marta Goleniowski Año: 2016 Facultad de Ciencias Agrarias y Forestales- Universidad Nacional de La Plata DEDICATORIA A Sandra, por confiar siempre en mí A mis padres, por los grandes valores que me han transmitido A Sergio, Luana, Juliana y Gonzalo, porque los amo y porque son lo mejor que me pasó en la vida A Eliana, por ser siempre mi gran compañera A Nelly, porque la sigo extrañando INDICE GENERAL AGRADECIMIENTOS i – ii ABREVIATURAS Y SIGLAS iii - vi TABLAS Y FIGURAS vii - xiii RESUMEN xiv - xvii ABSTRACT xviii - xxi I INTRODUCCION GENERAL……………….……………………………………….....1 II Objetivo General………………………………….……………………………….........6 III Objetivos Específicos……………………………………………………….…………6 IV Hipótesis…………………...……………………………………………….……….….7 V Bibliografía………………………………………………………………….………..7-12 CAPÍTULO I: Contexto teórico de la Investigación….…..…….……...........13-145 CAPÍTULO II: Propagación de las especies…...………….…………....…..146-279 CAPÍTULO III: Bioprospección………….……………………….………...….280-343 CONCLUSIONES…………….…………………………………………..………344-346 ANEXOS………………………………………………………………………….. 347 1 CAPITULO I: CONTEXTO TEÓRICO DE LA INVESTIGACIÓN .…………… 13 1.1 CONTEXTO REGIONAL Y PROVINCIAL…………………………………….. 13 1.1.1 La patagonia…………………………………………………………….. 13 1.1.2 Provincia de Río Negro……………………………………………….... 20 1.2 PROVINCIA FITOGEOGRÁFICA DEL MONTE………………….……. 22 1.2.1. Monte Pampeano o de Transición……………………………………. 25 1.3 DESARROLLO HISTÓRICO DE LA REGIÓN: LA DESERTIFICACIÓN EN LA PATAGONIA….................................................................................. 26 1.4 BIODIVERSIDAD, BIOTECNOLOGÍA Y BIOPROSPECCIÓN……..…… 29 1.4.1 Algunas Consideraciones Bioéticas………………………….............. 33 1.5 PROPAGACIÓN Y REPRODUCCIÓN DE LAS PLANTAS………….... 38 1.5.1 Propagación Sexual………………………………………………….. 39 1.5.1.1 Aspectos fisiológicos de la germinación……………...…..….. 39 1.5.1.2 Viabilidad, vigor, longevidad y conservación de semillas……. 40 viabilidad………….………...…………………………………...……. 41 vigor………………………...……………………………………...…….. 42 1.5.2 Propagación Vegetativa, Asexual o Clonal…………….………….… 43 1.5.2.1 Macropropagación…………………………………….………..…. 45 1.5.2.2. Micropropagación……………………...………………….….…... 46 i Cultivo de Tejidos Vegetales in vitro…………………………..……….. 46 ii Micropropagación……………………...…...….…………….................. 47 Principales métodos de micropropagación…………………………… 49 Aplicaciones de la Organogénesis…………………………………….. 52 Aplicaciones de la Embriogénesis Somática…………………………. 54 1.6 CULTIVO DE TEJIDOS DE ESPECIES LEÑOSAS…………………....... 55 1.6.1 Dificultades del CTV de especies leñosas…………………………… 57 1.7 METABOLISMO PRIMARIO, SECUNDARIO Y PRINCIPIOS BIOACTIVOS…………………………………………………………………....... 60 1.7.1 Características del metabolismo secundario…………………........... 60 1.7.2 Metabolitos secundarios en plantas de regiones áridas y semiáridas …………………………………………………..………………… 64 1.7.3 Bioprospección de metabolitos secundarios con valor nutricional…. 65 1.7.4 Caracterización físico química……………………………..…………. 67 1.7.4.1 Hidratos de carbono…………………………………………........ 67 1.7.4.2 Fibras……………………………………………………...……….. 68 1.7.4.3 Mucílagos………………………………………………….………. 68 1.7.4.4 Proteínas…………………………………………………………... 69 i Albuminas……………………………………………………………….... 71 ii Globulinas………………………………………………………………... 71 iii Prolaminas………………………………………………………………. 72 iv Glutelinas………………………………………………………………... 72 v Fraccionamiento de Proteínas…………………………………...…….. 73 vi Solubilidad de las Proteínas……………………………………………... 73 vii Efecto del pH en la solubilidad proteica………………………….….. 74 1.7.4.5 Lípidos……………………………………………………………... 74 i Ácidos grasos saturados………………………………..……………….. 75 ii Los ácidos grasos insaturados…………….…………….……………... 75 iii Ácidos grasos monoinsaturados………………………………..……….. 75 iv Ácidos grasos poliinsaturados………………………………….….…….. 76 1.7.4.6 Humedad……….……………………………………….……......... 76 1.7.4.7 Cenizas…………………………………………………………….. 76 1.8 ACTIVIDAD BIOLÓGICA…………………………………………...……… 77 1.8.1 Modelo del Pez Cebra…………………………………………...…….. 77 1.8.2. Determinación de Compuestos fenólicos…………………..……....... 78 1.8.3 Lectinas………………………………………………………..………… 79 1.8.4 Actividad Antioxidante………………………………………..………... 79 1.8.4.1 Método del DPPH.…………………………………………..…….. 80 1.9 LAS ESPECIES OBJETO DE ESTUDIO……………………………........ 81 1.9.1 El género Prosopis…………………………………………….….......... 81 1.9.2 Prosopis alpataco Phil. (Fabaceae, Mimosoideae).………………… 81 1.9.2.1 Ubicación taxonómica de la especie…………………….………. 81 1.9.2.2 Descripción Prosopis alpataco…...……………………………... 83 1.9.2.3 Usos populares de Prosopis…………………………….............. 87 i Usos alimenticios………………………………………………..………… 89 ii Usos farmacológicos…………………………………………..…………. 90 Principios activos………………………………………………………… 91 1.9.2.4 Usos no tradicionales de Prosopis………………….…………… 91 i Curtientes…………………………………………………………………... 91 ii Fibras...…………………………………………………………………….. 91 iii Tinturas……………………………….…………………………………… 91 iv Gomas……...…….……………………………………………….…...…. 92 v Miel……………………………………………………………………….… 92 vi Usos madereros (madera, leña y carbón)…………….…………….… 92 vii El valor ecológico de los Prosopis……………….…………..…...…… 94 1.9.3. El género Condalia………………………………………………….…. 95 1.9.4 Condalia microphylla Cav…………………………………….………... 96 1.9.4.1 Ubicación taxonómica de la especie…………………………….. 96 1.9.4.2 Descripción de Condalia microphylla……………………………. 96 1.9.4.3 Usos populares de la Familia Rhamnaceae y del género Condalia…………………………………………………………………..… 99 i Usos alimenticios……………………………………………………….... 99 ii Usos farmacológicos……………………………………………….….... 100 Principios activos……………………………………………………...... 100 1.9.4.4 Usos no tradicionales…………………………………………….. 101 i Tinturas…………………………………………………………………… 101 ii Insecticidas………………………………………………………………. 101 iii Miel……………………………………………………….………………… 101 iv Usos madereros…………………………………………………….…….. 102 1.10 BIBLIOGRAFIA…………………………….………………………….…… 102-145 2 CAPITULO II: PROPAGACIÓN DE LAS ESPECIES.……………………….. 146 2.1 PROPAGACIÓN DE Prosopis………………………………………….…… 146 2.1.1 Propagación sexual- Germinación de semillas de Prosopis………... 147 2.1.2 Propagación asexual o vegetativa de Prosopis………………………... 150 2.1.2.1 Macropropagación………………………………..……….……….… 150 2.1.2.2 Micropropagación…………………………………………………..... 151 2.2 PROPAGACIÓN DE Condalia Y OTRAS RHAMNACEAE........................ 153 2.2.1 Propagación sexual……….…………………………….…………........... 153 2.2.2 Germinación de Condalia microphylla…..………..…………………….. 157 2.2.3 Propagación asexual…………………………………..………….…….... 157 2.2.3.1 Macropropagación………………………………………..…………. 157 2.2.3.2 Micropropagación…………………………………………..……….. 158 2.3 OBJETIVOS…………………………….……………………………………..... 160 2.4 MATERIALES Y MÉTODOS……………………………………………….…. 160 2.4.1 Germinación………………………….…………………………………..... 160 2.4.1.1 Recolección de semillas………………………………………….… 160 2.4.1.2 Plantas Madres…………………………………………………….… 164 2.4.1.3 Cosecha de las semillas………………………………………….… 164 2.4.1.4 Determinación de la fecha de recolección idónea……………… 165 2.4.1.5 Modalidad de cosecha …………………………………….….... 165 2.4.1.6 Procesamiento y manejo de las semillas……………………….… 165 2.4.1.7 Evaluación de la germinación………………………………….….. 168 2.4.1.8 Tratamiento estadístico de los datos………………………….…… 170 2.4.2 Germinación de Prosopis alpataco……………………………………... 170 2.4.2.1 Condiciones ambientales…………………….……………………. 170 2.4.2.2 Determinación de la temperatura óptima…………………………. 171 2.4.2.3 Dormancia…………………………………….……………………... 172 2.4.2.4 Evaluación de las plántulas obtenidas………………………….... 173 2.4.2.5 Patrón de imbibición de las semillas (a 30ºC).…………………... 173 2.4.2.6 Viabilidad………………………………………………….…………. 174 2.4.2.7 Vigor (métodos indirectos).………………………………………… 175 i Prueba de conductividad eléctrica……………..…………………………. 175 ii Prueba colorimétrica……………………………….……………………… 176 2.4.3 Germinación de Condalia microphylla…….……………………............ 176 2.4.3.1 Escarificación mecánica con papel de lija………………….……. 178 2.4.3.2 Escarificación mecánica con pinza tipo pelacable……………… 178 2.4.3.3 Escarificación de endocarpos con H2SO4 concentrado.……….. 178 2.4.3.4 Inmersión en agua……………….…………………………………. 178 2.4.3.5 Estratificación fría y húmeda………….…………………………… 178 2.4.3.6 Germinación de semillas de C. microphylla en invernadero….... 179 2.4.3.7 Germinación de semillas de C. microphylla en condiciones de 179 laboratorio………………………………………………………….…….…… 2.4.3.8 Evaluación de la imbibición en los endocarpos……………….... 180 2.4.4 Micropropagación de P. alpataco………………………………………. 180 2.4.4.1 Acondicionamiento y desinfección de las semillas………….… 180 2.4.4.2 Medios de cultivo y reguladores de crecimiento……………...… 180 2.4.4.3 Germinación in vitro………………………………………………... 182 2.4.4.4 Selección del explante…………………………………………….. 182 2.4.4.5 Establecimiento in vitro……………………………………………. 183 i Enraizamiento in vitro (organogénesis)..………………………………... 183 ii Inducción de brotes (organogénesis)……………………………………. 183 iii Embriogénesis somática………………………………………………..... 183 Determinación de la producción de embriones………………….……... 184 Germinación de los embriones somáticos………………………………. 184 2.4.4.6 Condiciones ambientales…………………………………….….… 185 2.4.4.7 Aclimatación y rusticación de plantas obtenidas vía 185 organogénesis y embriogénesis somática…………………………….…. 2.4.4.8 Análisis de las respuestas morfogénicas………………………... 185 2.4.4.9 Cortes con micrótomo…………………………………………….... 186 2.5 RESULTADOS…………………………….…………….…………………….. 187 2.5.1
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