Caracterización de la Diversidad Microbiana de Suelos Destinados a Cultivos de Ananas comosus (piña), Tratados con Distintos Plaguicidas en el Municipio de Lebrija, Santander

Digital

Autores:: Bravo Granados, Natalia Andrea

Tipo de recurso:: Trabajo de grado de pregrado

Fecha de publicación:: 2021

Institución:: Universidad de Santander

Repositorio:: Repositorio Universidad de Santander

Idioma:: spa

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dc.title.spa.fl_str_mv	Caracterización de la Diversidad Microbiana de Suelos Destinados a Cultivos de Ananas comosus (piña), Tratados con Distintos Plaguicidas en el Municipio de Lebrija, Santander
title	Caracterización de la Diversidad Microbiana de Suelos Destinados a Cultivos de Ananas comosus (piña), Tratados con Distintos Plaguicidas en el Municipio de Lebrija, Santander
spellingShingle	Caracterización de la Diversidad Microbiana de Suelos Destinados a Cultivos de Ananas comosus (piña), Tratados con Distintos Plaguicidas en el Municipio de Lebrija, Santander Metagenómica Piña ADN Secuenciación Metagenomics Pineapple DNA IlluminaMiSeq
title_short	Caracterización de la Diversidad Microbiana de Suelos Destinados a Cultivos de Ananas comosus (piña), Tratados con Distintos Plaguicidas en el Municipio de Lebrija, Santander
title_full	Caracterización de la Diversidad Microbiana de Suelos Destinados a Cultivos de Ananas comosus (piña), Tratados con Distintos Plaguicidas en el Municipio de Lebrija, Santander
title_fullStr	Caracterización de la Diversidad Microbiana de Suelos Destinados a Cultivos de Ananas comosus (piña), Tratados con Distintos Plaguicidas en el Municipio de Lebrija, Santander
title_full_unstemmed	Caracterización de la Diversidad Microbiana de Suelos Destinados a Cultivos de Ananas comosus (piña), Tratados con Distintos Plaguicidas en el Municipio de Lebrija, Santander
title_sort	Caracterización de la Diversidad Microbiana de Suelos Destinados a Cultivos de Ananas comosus (piña), Tratados con Distintos Plaguicidas en el Municipio de Lebrija, Santander
dc.creator.fl_str_mv	Bravo Granados, Natalia Andrea
dc.contributor.advisor.none.fl_str_mv	Rueda Forero, Nohora Juliana Suárez Barrera, Miguel Orlando
dc.contributor.author.none.fl_str_mv	Bravo Granados, Natalia Andrea
dc.subject.proposal.spa.fl_str_mv	Metagenómica Piña ADN Secuenciación Metagenomics Pineapple DNA
topic	Metagenómica Piña ADN Secuenciación Metagenomics Pineapple DNA IlluminaMiSeq
dc.subject.proposal.eng.fl_str_mv	IlluminaMiSeq
description	Digital
publishDate	2021
dc.date.accessioned.none.fl_str_mv	2021-08-21T01:16:20Z
dc.date.available.none.fl_str_mv	2021-08-21T01:16:20Z
dc.date.issued.none.fl_str_mv	2021-05-06
dc.type.spa.fl_str_mv	Trabajo de grado - Pregrado
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dc.identifier.local.none.fl_str_mv	T 33.21 B719c
dc.identifier.uri.none.fl_str_mv	https://repositorio.udes.edu.co/handle/001/5530
identifier_str_mv	T 33.21 B719c
url	https://repositorio.udes.edu.co/handle/001/5530
dc.language.iso.spa.fl_str_mv	spa
language	spa
dc.rights.spa.fl_str_mv	Derechos Reservados - Universidad de Santander, 2021
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dc.rights.creativecommons.spa.fl_str_mv	Atribución-NoComercial-SinDerivadas 4.0 Internacional (CC BY-NC-ND 4.0)
dc.rights.uri.spa.fl_str_mv	https://creativecommons.org/licenses/by-nc-nd/4.0/
rights_invalid_str_mv	Derechos Reservados - Universidad de Santander, 2021 Atribución-NoComercial-SinDerivadas 4.0 Internacional (CC BY-NC-ND 4.0) https://creativecommons.org/licenses/by-nc-nd/4.0/ http://purl.org/coar/access_right/c_abf2
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dc.format.extent.spa.fl_str_mv	159 p
dc.format.mimetype.spa.fl_str_mv	application/pdf
dc.publisher.spa.fl_str_mv	Bucaramanga : Universidad de Santander, 2021
dc.publisher.faculty.spa.fl_str_mv	Facultad de Ciencias Exactas, Naturales y Agropecuarias
dc.publisher.place.spa.fl_str_mv	Bucaramanga, Colombia
dc.publisher.program.spa.fl_str_mv	Microbiología Industrial
institution	Universidad de Santander
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spelling	Rueda Forero, Nohora Julianafdefebdb-5a74-4c59-ba33-4ff4cc09bca4-1Suárez Barrera, Miguel Orlando335becb1-2371-418a-acf8-21265141ac00-1Bravo Granados, Natalia Andrea16e6a42d-caeb-4028-9efd-962d5556bec1-12021-08-21T01:16:20Z2021-08-21T01:16:20Z2021-05-06DigitalEl suelo es un hábitat diverso, su composición varía dependiendo del uso, condiciones climáticas y actividades antropogénicas; y conocer los microorganismos presentes requiere la implementación de herramientas de secuenciación de nueva generación. Ananás comosus es un cultivo de gran importancia para Santander, sin embargo, los estudios referentes a la caracterización biológica del suelo asociado, son escasas en la región y en el país. En ese sentido, este trabajo se centró en la caracterización microbiana del suelo destinado a un cultivo de piña. Para lograrlo, las muestras se tomaron aleatoriamente de cuatro lotes a 20 cm de profundidad, obteniendo un total de 14 muestras. Se estandarizó el protocolo de extracción de ADN genómico total de suelo y la cantidad de ADN de cada muestra fue >60 ng/μl. Posteriormente se enviaron a secuenciar a MacroGen mediante la plataforma Illumina MiSeq (2x300 pb), obteniéndose un total de 1’518.101 secuencias de las muestras analizadas. El empleo de herramientas metagenómicas como el FASTQC y el MG-RAST, permitieron establecer la calidad de las secuencias obtenidas y la clasificación desde dominio hasta género de los microorganismos presentes en los suelos estudiados. Filo como Actinobacteria (>50%), Acidobacteria (>18%), Firmicutes (>16%) y Verrumicrobia (>10%), se encontraron con mayor predominancia entre las 14 muestras estudiadas. A nivel de género se encontraron en abundancia Candidatus Koribacter, Streptomyces, Isosphaera, Bacillus, Candidatus Solibacter y Actinomadura. Con respecto a la diversidad α, esta varía por las condiciones nutricionales del suelo, es por esto que en la muestra del suelo Blanco los valores de esta diversidad son los más bajos. Finalmente, se evidenció elevada similitud entre la composición de las comunidades microbianas de los cuatro suelos en estudio, indicando que el plaguicida y el bioplaguicida, no tienen un efecto significativo en la composición de la población microbiana presentes en la rizosfera del cultivo de piña.The soil is a diverse habitat, its composition varies depending on the use, climatic conditions and anthropogenic activities; and knowing the groups of microorganisms present requires the implementation of next-generation sequencing tools. Ananas comosus is a crop of great importance for Santander, however, studies referring to the biological characterization of the associated soil are scarce in the region and the country. In this sense, this work focused on the microbial characterization of the soil destined for a pineapple crop. The samples were randomly taken from four batches at a depth of 20 cm, obtaining a total of 14 samples. The total genomic soil DNA extraction protocol was standardized and the amount of DNA in each sample was> 60 ng / μl. Subsequently, they were sent to be sequenced to MacroGen using the Illumina MiSeq platform, obtaining a total of 1,518,101 sequences of the analyzed samples. The use of metagenomic tools such as FASTQC and MG-RAST made it possible to establish the quality of the sequences obtained and the classification from domain to the gender of the microorganisms present in the soils studied. Phylum such as Actinobacteria (> 50%), Acidobacteria (> 18%), Firmicutes (> 16%) and Verrumicrobia (> 10%), were found to be more prevalent among the 14 samples studied. At the genus level, Candidatus Koribacter, Streptomyces, Isosphaera, Bacillus, Candidatus Solibacter and Actinomadura were found in abundance. Concerning the diversitPregradoMicrobiólogo Industrial1 ed.Introducción ..............................................................................................................................15 Marco Teórico ...........................................................................................................................18 El Suelo .................................................................................................................................18 Aspectos Físicos del Suelo ................................................................................................19 Aspectos Químicos del Suelo ............................................................................................21 Materia Orgánica ...............................................................................................................23 La Agricultura .....................................................................................................................24 Microbiota del Suelo ..............................................................................................................25 Microorganismos de la Rizosfera .......................................................................................25 Funciones de los Microorganismos ....................................................................................26 Secuenciación de Nueva Generación ....................................................................................28 NGS en la Agricultura ........................................................................................................32 Illumina ..............................................................................................................................33 Análisis Bioinformático de Secuencias ..................................................................................39 MG-RAST como Herramienta Bioinformática .....................................................................41 Análisis Estadístico................................................................................................................43 Past Software ....................................................................................................................44 Estado del Arte .........................................................................................................................46 Marco Legal ..............................................................................................................................50 Planteamiento y Justificación del Problema ..............................................................................51 Pregunta de investigación .........................................................................................................52 Hipótesis ...................................................................................................................................53 Objetivos ...................................................................................................................................54 Objetivo General ...................................................................................................................54 Objetivos Específicos ............................................................................................................54 Metodología ..............................................................................................................................55 Especificaciones del Cultivo de Piña .....................................................................................55 Muestreo ...............................................................................................................................56 Extracción de ADN Genómico ...............................................................................................57 Amplificación y Secuenciación del Gen Ribosomal 16S. .......................................................58 Análisis Bioinformáticos .........................................................................................................58 Análisis Estadísticos ..............................................................................................................59 Análisis Físico-Químicos (realizados por los estudiantes de la UPB, sede Bucaramanga) ....59 Resultados y Análisis de Resultados ........................................................................................60 Extracción y Cuantificación de ADN Metagenómico ..............................................................60 Análisis Bioinformático de Secuencias ..................................................................................61 Clasificación por Dominios .................................................................................................61 Clasificación por filo ...........................................................................................................63 Clasificación por Género ....................................................................................................65 Diversidad α .......................................................................................................................67 Abundancia Relativa de Géneros Dominantes ...................................................................68 Géneros Únicos y Comunes en los Tratamientos ..............................................................69 Géneros predominantes .....................................................................................................71 Análisis Estadísticos ..............................................................................................................73 Análisis de Componentes Principales (ACP) ......................................................................73 Diversidad β .......................................................................................................................77 Discusión ..................................................................................................................................78 Análisis Fisicoquímicos .........................................................................................................78 Propiedades Físicas del Suelo ...........................................................................................78 Propiedades Químicas del Suelo .......................................................................................78 Extracción de ADN ................................................................................................................81 Análisis Bioinformático de Secuencias ..................................................................................83 Clasificación por Dominio ...................................................................................................84 Clasificación por filo ...........................................................................................................85 Clasificación por Género ....................................................................................................91 Diversidad α ..................................................................................................................... 101 Abundancia Relativa y Efecto del Plaguicida (Lorsban) y Bioplaguicida (Nicotina y Capsaicina). ..................................................................................................................... 102 Géneros Únicos y Comunes en los Tratamientos ................................................................ 104 Análisis de Componentes Principales (ACP) ....................................................................... 106 Diversidad β ........................................................................................................................ 107 Conclusiones .......................................................................................................................... 109 Recomendaciones .................................................................................................................. 111 Referencias Bibliográficas ....................................................................................................... 112 Apéndices ............................................................................................................................... 149 Apendice A. Protocolo de Extracción de ADN Estandarizado: Kit E.Z.N.A.® Soil DNA Kit (OMEGA) ............................................................................................................................ 149 Apéndice B. Concentración de ADN Genómico Total de Suelos ......................................... 151 Apéndice C. Calidad del ADN Extraído ................................................................................ 152 Apéndice D. Secuencias Obtenidas por Muestra ................................................................. 153 Apéndice E. Curva de Rarefacción de los Suelos Analizados ............................................. 154 Apéndice F. Prueba de Kruskal Wallis – Composición microbiana sin diferencia significativa ............................................................................................................................................ 156 Apéndice G. Propiedades físicoquímicas de los suelos estudiados, realizados por los estudiantes de la Universidad Pontificia Bolivariana sede Bucaramanga ............................ 157 Cálculo de Nitrógeno de mg NTK/Kg a %. ........................................................................... 159159 papplication/pdfT 33.21 B719chttps://repositorio.udes.edu.co/handle/001/5530spaBucaramanga : Universidad de Santander, 2021Facultad de Ciencias Exactas, Naturales y AgropecuariasBucaramanga, ColombiaMicrobiología IndustrialDerechos Reservados - Universidad de Santander, 2021info:eu-repo/semantics/openAccessAtribución-NoComercial-SinDerivadas 4.0 Internacional (CC BY-NC-ND 4.0)https://creativecommons.org/licenses/by-nc-nd/4.0/http://purl.org/coar/access_right/c_abf2MetagenómicaPiñaADNSecuenciaciónMetagenomicsPineappleDNAIlluminaMiSeqCaracterización de la Diversidad Microbiana de Suelos Destinados a Cultivos de Ananas comosus (piña), Tratados con Distintos Plaguicidas en el Municipio de Lebrija, SantanderTrabajo de grado - Pregradohttp://purl.org/coar/resource_type/c_7a1fTextinfo:eu-repo/semantics/bachelorThesishttps://purl.org/redcol/resource_type/TPhttp://purl.org/coar/version/c_71e4c1898caa6e32Todas las AudienciasAguirre V., Delgado, V. 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Antonie Van Leeuwenhoek 108:1331–1339.PublicationLICENSElicense.txtlicense.txttext/plain; charset=utf-859https://repositorio.udes.edu.co/bitstreams/d7cdff84-8f91-4c31-b9aa-bd33404cf4a6/download38d94cf55aa1bf2dac1a736ac45c881cMD52ORIGINALCaracterización de la diversidad microbiana de suelos destinados a cultivos de Ananas comosus (piña)....pdfCaracterización de la diversidad microbiana de suelos destinados a cultivos de Ananas comosus (piña)....pdfDocumento Principalapplication/pdf2311935https://repositorio.udes.edu.co/bitstreams/0af88ba0-af8f-4481-834f-2424a443d381/downloadb146a3146015420b56565d787c6b70fbMD51TEXTCaracterización de la diversidad microbiana de suelos destinados a cultivos de Ananas comosus (piña)....pdf.txtCaracterización de la diversidad microbiana de suelos destinados a cultivos de Ananas comosus (piña)....pdf.txtExtracted texttext/plain101496https://repositorio.udes.edu.co/bitstreams/3f63e337-9435-4c4b-ba89-fabdfa52a06e/downloadee5cef9a2796f4193e75115dc39a25cdMD53THUMBNAILCaracterización de la diversidad microbiana de suelos destinados a cultivos de Ananas comosus (piña)....pdf.jpgCaracterización de la diversidad microbiana de suelos destinados a cultivos de Ananas comosus (piña)....pdf.jpgGenerated Thumbnailimage/jpeg5935https://repositorio.udes.edu.co/bitstreams/d5b00a00-210e-409e-a72c-15fe39ed462c/download8b1a7e2f877fb9f7e259b6dc8a91d1ecMD54001/5530oai:repositorio.udes.edu.co:001/55302022-10-25 10:11:43.436https://creativecommons.org/licenses/by-nc-nd/4.0/Derechos Reservados - Universidad de Santander, 2021https://repositorio.udes.edu.coRepositorio Universidad de Santandersoporte@metabiblioteca.comTGljZW5jaWEgZGUgUHVibGljYWNpw7NuIFVERVMKRGlyZWN0cmljZXMgZGUgVVNPIHkgQUNDRVNPCgo=

Caracterización de la Diversidad Microbiana de Suelos Destinados a Cultivos de Ananas comosus (piña), Tratados con Distintos Plaguicidas en el Municipio de Lebrija, Santander

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