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(Azotobacter Y Rhizobium) Para Tarwi Y Frijol Caupí Como Alternativa Ambiental a La Fertilización Nitrogenada

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(Azotobacter Y Rhizobium) Para Tarwi Y Frijol Caupí Como Alternativa Ambiental a La Fertilización Nitrogenada Universidad Nacional Mayor de San Marcos Universidad del Perú. Decana de América Dirección General de Estudios de Posgrado Facultad de Ingeniería Geológica, Minera, Metalúrgica y Geográfica Unidad de Posgrado Evaluación de un biofertilizante (Azotobacter y Rhizobium) para tarwi y frijol caupí como alternativa ambiental a la fertilización nitrogenada TESIS Para optar el Grado Académico de Magíster en Ciencias Ambientales con mención en Gestión y Control de la Contaminación AUTOR Erika Yovana GONZALES MEDINA ASESOR Mg. Mario ALCARRAZ CURI Lima, Perú 2019 Reconocimiento - No Comercial - Compartir Igual - Sin restricciones adicionales https://creativecommons.org/licenses/by-nc-sa/4.0/ Usted puede distribuir, remezclar, retocar, y crear a partir del documento original de modo no comercial, siempre y cuando se dé crédito al autor del documento y se licencien las nuevas creaciones bajo las mismas condiciones. No se permite aplicar términos legales o medidas tecnológicas que restrinjan legalmente a otros a hacer cualquier cosa que permita esta licencia. Referencia bibliográfica Gonzales, E. (2019). Evaluación de un biofertilizante (Azotobacter y Rhizobium) para tarwi y frijol caupí como alternativa ambiental a la fertilización nitrogenada. Tesis para optar grado de Magíster en Ciencias Ambientales con mención en Gestión y Control de la Contaminación. Unidad de Posgrado, Facultad de Ingeniería Geológica, Minera, Metalúrgica y Geográfica, Universidad Nacional Mayor de San Marcos, Lima, Perú. HOJA DE METADATOS COMPLEMENTARIOS CODIGO ORCID DEL AUTOR: 0000-0001-9172-0158 CODIGO ORCID DEL ASESOR: 0000-0001-5262-2969 DNI: 41835178 GRUPO DE INVESTIGACIÓN: Grupo de investigación sobre Microbiología Industrial, en el laboratorio de Bioprocesos Industriales de la Universidad Nacional Mayor de San Marcos (UNMSM). Grupo de investigación en Biofertilizantes en la Universidad San Ignacio de Loyola (USIL). INSTITUCIÓN QUE FINANCIA PARCIAL O TOTALMENTE LA INVESTIGACIÓN: Autofinanciado UBICACIÓN GEOGRÁFICA DONDE SE DESARROLLÓ LA INVESTIGACIÓN. DEBE INCLUIR LOCALIDADES Y COORDENADAS GEOGRÁFICAS El lugar de muestreo se realizó en Lima, distrito La Molina, Perú. Las coordenadas geográficas: latitud: 12° 05´ 06´´ S, longitud 76° 57´ 09´´ S.O, una altura de 236 m.s.n.m. AÑO O RANGO DE AÑOS QUE LA INVESTIGACIÒN ABARCÓ: Agosto 2017- Marzo 2018 DEDICATORIA La presente tesis se la dedico en primer lugar a Dios, por darme mucha la fortaleza, fe y esperanza durante la realización de la tesis, a todos mis seres queridos gracias por acompañarme y brindarme su apoyo, comprensión y aliento. - 2 - AGRADECIMIENTOS Le agradezco mucho a Dios por darme la oportunidad de culminar mi tesis, por darme la fortaleza que necesito, por darme la paz y tranquilidad en todo momento. Gracias Sr. por bendecirme y poner en mi camino tan buenas personas que me acompañaron en la realización de la tesis. A mi querida Madre, de quien estoy muy orgullosa, gracias mamita por siempre estar allí cuando te necesitaba para alentarme y apoyarme. Me siento muy feliz y siempre te llevo en mi corazón. A mi padre Carlitos, gracias papá por tus consejos y apoyo constante. A mis hermanos Carlos y Karina, de la misma muchas gracias hermanos por darme su apoyo y comprensión siempre. A Manuel Hallasi Vega, por su constante apoyo durante la realización de la tesis. Sus palabras motivadoras, su comprensión, su gran apoyo en todo este tiempo. A todo equipo de Investigación del Laboratorio de Bioprocesos de la Universidad Nacional Mayor de San Marcos, al Prof. Mg. Sc. Mario Alcarraz Curi, por todos sus consejos y gracias a todos los compañeros del laboratorio, en especial a Katty y Anita, Raúl y Julián por apoyo constante y compañerismo durante la realización de la tesis. A mis compañeros de mi centro de Labores la Universidad San Ignacio de Loyola, por su apoyo durante la realización de la tesis. A los miembros del jurado por el tiempo dedicado a la revisión de mi proyecto de tesis y borrador de tesis. Por sus recomendaciones y sugerencias para la realización del trabajo de investigación. - 3 - TABLA DE CONTENIDO CAPITULO 1. INTRODUCCIÓN .................................................................................................... 10 1.1 Situación Problemática ....................................................................................................................... 11 1.2 Formulación del Problema ................................................................................................................ 13 1.3 Justificación teórica ............................................................................................................................ 14 1.4 Justificación práctica .......................................................................................................................... 15 1.5 Objetivos .............................................................................................................................................. 18 CAPÍTULO 2: MARCO TEÓRICO ................................................................................................. 19 2.1. Antecedentes de investigación ....................................................................................................... 19 2.1.1 Fertilizantes nitrogenados y sus formas contaminantes 19 2.1.2 Impactos negativos de los fertilizantes 20 2.1.3. Biofertilización como alternativa de reducción de la contaminación por nitrógeno 22 2.1.4. Microorganismos promotores de crecimiento vegetal y mecanismos de acción. 24 2.1.5. Aplicación de los biofertilizantes en Sudamérica 25 2.2 Bases teóricas .................................................................................................................................... 31 2.2.1 Clasificación taxonomía y características de Rhizobium 31 2.2.2 Rhizobium como biofertilizante 33 2.2.3 Rhizobium como promotor de crecimiento vegetal PGPR 34 2.2.4 Clasificación taxonómica y características de Azotobacter 36 2.2.5 Azotobacter como biofertilizante 39 2.2.6 El cultivo del tarwi (Lupinus mutabilis) 40 2.2.7 El cultivo de frijol caupí (Vigna unguiculata) 44 2.2.8. Aplicaciones del cultivo 47 2.3 Marco conceptual o glosario ............................................................................................................. 48 2.3.1 Azotobacter 48 2.3.2 Rhizobium 48 2.3.3 Agricultura sustentable 48 2.3.4 Biofertilizantes 49 2.3.5 Fijación biológica de nitrógeno (FBN) 49 2.3.6. Eutrofización 50 2.3.7. Las bacterias promotoras de crecimiento (PGPRs) 50 - 4 - CAPITULO 3: MATERIALES Y MÉTODOLOGÍA ....................................................................... 51 3.1 MATERIALES ..................................................................................................................................... 51 3.1.1 Material biológico 51 3.1.2 Medios de cultivo 51 3.1.3 Soluciones y reactivos 51 3.1.4 Materiales y equipos de laboratorio 52 3.1.5. Materiales para el ensayo en bandejas 53 3.1.6. Materiales para la elaboración del invernadero 53 3.2 MÉTODOLOGÍA ................................................................................................................................. 54 3.2.1 Lugar de toma de muestras y toma de muestras de suelo y plántulas 54 3.2.2 Aislamiento de Rhizobium 56 3.2.3 Aislamiento de Azotobacter 57 3.2.4 Tinción de Gram 58 3.2.5 Tinción de flagelos 59 3.2.6 Preparación de los inoculantes 59 3.2.7 Experimento en bandejas con semillas de caupí 61 3.2.8 Experimento en invernadero con semillas de caupí 62 3.2.9 Experimento en bandejas con semillas de tarwi (Lupinus mutabilis) 64 3.2.10 Experimento en invernadero con semillas de tarwi 64 CAPITULO 4. RESULTADOS ....................................................................................................... 65 4.1. Evaluación de las plántulas muestreadas ..................................................................................... 65 4.2. Caracterización de las cepas de Rhizobium ................................................................................. 66 4.3 Caracterización de las cepas de Azotobacter sp .......................................................................... 69 4.4 Observación de las células por medio de la Tinción de GRAM .................................................. 71 4.5 Resultado de la tinción de flagelos con el reactivo Leifson ......................................................... 72 4.6 Evaluación de la germinación de las semillas de caupí ............................................................... 73 4.7. Evaluación de la presencia de hongos en bandejas ................................................................... 78 4.8 Resultados del experimento de caupí en invernadero ................................................................. 79 CAPITULO 5. DISCUSIONES ....................................................................................................... 97 5.1 Aislamiento e identificación de colonias de Rhizobium ................................................................ 97 5.2 Tinción Gram de colonias de Rhizobium ........................................................................................ 97 5.3. Aislamiento e identificación de colonias de Azotobacter ............................................................ 98 5.4. Tinción Gram y de flagelos de Azotobacter .................................................................................. 99 5.5 Efecto de las cepas individuales (Rhizobium o Azotobacter) en la germinación de semillas de frijol caupí
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