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ACUMULACION DE TREALOSA EN GENOTIPOS DE FRIJOL SOMETIDOS a SEQUIA Para Obtener El Grado De UNIVERSIDAD MICHOACANA DE SAN NICOLAS DE HIDALGO Instituto de Investigaciones Químico – Biológicas Programa de doctorado Institucional en Ciencias Biológicas, Opción en Biología Experimental TESIS ACUMULACION DE TREALOSA EN GENOTIPOS DE FRIJOL SOMETIDOS A SEQUIA Para obtener el grado de: DOCTORA EN CIENCIAS EN BIOLOGIA EXPERIMENTAL Presenta: M.C: MARIA DE LOURDES BALLESTEROS ALMANZA DIRECTOR DE TESIS: DR. RODOLFO FARIAS RODRIGUEZ CO-DIRECTOR: DR. JUAN JOSE PEÑA CABRIALES MORELIA, MICH, MEXICO Mayo, 2010 1 CONTENIDO I.RESUMEN .................................................................................................................... 8 ABSTRACT ..................................................................................................................... 9 II. INTRODUCCIÓN ....................................................................................................... 10 III. ANTECEDENTES ...................................................................................................... 13 III.1. LEGUMINOSAS ............................................................................................................ 13 III.2. ORIGEN Y DOMESTICACIÓN DEL FRIJOL COMÚN: RAZAS Y CLASES COMERCIALES........... 14 III.2.1. Importancia del frijol .................................................................................................................... 18 III.3. DIVERSIDAD GENÉTICA EN LAS ESPECIES DE Phaseolus EN RELACIÓN CON EL FRIJOL COMÚN. ............................................................................................................................. 19 III.4. SIMBIOSIS Rhizobium/LEGUMINOSA ............................................................................. 21 III.5. MECANISMOS DE LAS PLANTAS A FACTORES ADVERSOS .............................................. 23 III.6. TREALOSA ................................................................................................................... 24 III.6.1. Propiedades químicas de la trealosa ............................................................................................ 25 III.6.2. Trealosa en la Naturaleza ............................................................................................................. 26 III.6.3. Trealosa en Microorganismos....................................................................................................... 27 III.6.4. Trealosa en las plantas .................................................................................................................. 28 III.6.5. Sintesis de trealosa en sistemas simbióticos ................................................................................ 29 III.6.6. Mecanismo de acción de la trealosa ............................................................................................. 34 III.6.7. La trealosa en la tolerancia al estrés hídrico en las plantas de frijol ............................................ 36 IV. HIPOTESIS .............................................................................................................. 38 V. OBJETIVOS .............................................................................................................. 39 VI. MATERIALES Y METODOS ....................................................................................... 40 VI.1. MATERIAL BIOLÓGICO ................................................................................................. 40 VI.2. PREPARACIÓN DE INOCULANTES ................................................................................. 40 VI.3. SEMILLAS Y DESINFECCIÓN .......................................................................................... 41 VI.4. SUSTRATO Y DESINFECCIÓN ........................................................................................ 41 VI.5. SIEMBRA E INOCULACIÓN DE SEMILLAS ....................................................................... 42 VI.6. CONDICIONES DE RIEGO .............................................................................................. 42 VI.7. DETERMINACIÓN DE LA ACUMULACIÓN DE TREALOSA ................................................. 42 2 VI.8. EXTRACCIÓN DE AZÚCARES SOLUBLES ......................................................................... 44 VI.9. CUANTIFICACIÓN DE TREALOSA ................................................................................... 45 VI.10. DETERMINACIÓN DE PARÁMETROS AGRONÓMICOS .................................................. 47 VI.11. DETERMINACIÓN DEL PORCENTAJE DE MICORRIZACIÓN EN RAÍCES ............................ 48 VI.12. PREPARACIÓN DE MUESTRAS PARA MICROSCOPÍA ELECTRÓNICA DE BARRIDO (MEB) 48 VI.13. DISEÑO EXPERIMENTAL ............................................................................................. 48 VI.14. ANÁLISIS ESTADÍSTICO .............................................................................................. 51 VII. RESULTADOS ......................................................................................................... 52 VII.1. ACUMULACIÓN DE TREALOSA EN PLANTAS DE FRIJOL SOMETIDAS A ESTRÉS HÍDRICO . 52 VII.1.1. Trealosa en nódulos .................................................................................................................... 52 VII.1.1.1. Phaseolus vulgaris P1325687 ............................................................................................... 52 VII.1.1.2. Phaseolus vulgaris Flor de Mayo M38 ................................................................................. 53 VII.1.1.3. Phaseolus acutifolius P40142 ............................................................................................... 54 VII.1.1.4. Phaseolus vulgaris cv. Pinto Saltillo ..................................................................................... 55 VII.1.1.5. Phaseolus filiformis p1535309 ............................................................................................. 56 VII.1.2. Acumulación de trealosa en raíz ................................................................................................. 57 VII.1.3. Trealosa en tallo .......................................................................................................................... 59 VII.1.4. Trealosa en hojas ......................................................................................................................... 59 VII.1.5. Acumulación de trealosa en semilla ............................................................................................ 60 VII.2. ÁREA FOLIAR DE FRIJOL SOMETIDO A DIFERENTES TRATAMIENTOS DE INOCULACIÓN Y ESTRÉS HÍDRICO .................................................................................................................. 62 VII.3. BIOMASA DE GENOTIPOS DE FRIJOL SOMETIDO A SEQUÍA Y BAJO DIFERENTES TRATAMIENTOS DE INOCULACIÓN ...................................................................................... 64 VII.4. CONTENIDO RELATIVO DE AGUA (CRA) EN HOJAS DE FRIJOL SOMETIDO A DIFERENTES TRATAMIENTOS DE INOCULACIÓN ...................................................................................... 66 VII.4.1. Phaseolus vulgaris P132568 ........................................................................................................ 66 VII.4.2. Phaseolus vulgaris cv. Flor de Mayo M38 ................................................................................... 67 VII.4.3. Phaseolus acutifolius G40142 ...................................................................................................... 68 VII.4.4. Phaseolus vulgaris cv. Pinto Saltillo ............................................................................................. 69 VII.4.5. Phaseolus filiformis P1535309 ..................................................................................................... 70 VII.5. PESO FRESCO DE NÓDULOS DE GENOTIPOS DE FRIJOL BAJO DIFERENTES TRATAMIENTOS DE INOCULACIÓN ...................................................................................... 71 VII.5.1. Phaseolus vulgaris P132568 ........................................................................................................ 71 VII.5.2. Phaseolus vulgaris cv. Flor de Mayo ............................................................................................ 72 VII.5.3. Phaseolus acutifolius G40142 ...................................................................................................... 73 VII.5.4. Phaseolus vulgaris cv. Pinto Saltillo ............................................................................................. 74 VII.5.5. Phaseolus filiformis P1535309 ..................................................................................................... 75 3 VII.6. PESO SECO DE NÓDULO DE FRIJOL SOMETIDO A DIFERENTES TRATAMIENTOS DE INOCULACIÓN BAJO ESTRÉS HIDRICO .................................................................................. 76 VII.6.1. Phaseolus vulgaris P132568 ........................................................................................................ 76 VII.6.2. Phaseolus vulgaris cv. Flor de Mayo ............................................................................................ 77 VII.6.3. Phaseolus acutifolius P132568 .................................................................................................... 78 VII.6.4. Phaseolus vulgaris cv. Pinto Saltillo ............................................................................................. 79 VII.6.5. Phaseolus filiformis P1535309 ....................................................................................................
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