Evaluación de la microbiota del tracto gastrointestinal del biotipo de maíz de Spodoptera frugiperda en presencia de endotoxinas del Bacillus thuringiensis
ilustraciones, diagramas, tablas
- Autores:
-
Castañeda Molina, Yuliana del Pilar
- Tipo de recurso:
- Fecha de publicación:
- 2021
- Institución:
- Universidad Nacional de Colombia
- Repositorio:
- Universidad Nacional de Colombia
- Idioma:
- spa
- OAI Identifier:
- oai:repositorio.unal.edu.co:unal/82152
- Palabra clave:
- 570 - Biología::577 - Ecología
570 - Biología::576 - Genética y evolución
570 - Biología::572 - Bioquímica
Maíz - Enfermedades y plagas
Spodoptera frugiperda
Arsenophonus
Biotipo maíz
Endotoxinas BT
Microbiota intestinal
Resistencia
Corn Strain
Bt toxins
Gut microbiota
Resistance
- Rights
- openAccess
- License
- Reconocimiento 4.0 Internacional
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| dc.title.spa.fl_str_mv |
Evaluación de la microbiota del tracto gastrointestinal del biotipo de maíz de Spodoptera frugiperda en presencia de endotoxinas del Bacillus thuringiensis |
| dc.title.translated.eng.fl_str_mv |
Evaluation of Spodoptera frugiperda (corn strain) gut microbiota in presence of Bacillus thuringiensis endotoxins |
| title |
Evaluación de la microbiota del tracto gastrointestinal del biotipo de maíz de Spodoptera frugiperda en presencia de endotoxinas del Bacillus thuringiensis |
| spellingShingle |
Evaluación de la microbiota del tracto gastrointestinal del biotipo de maíz de Spodoptera frugiperda en presencia de endotoxinas del Bacillus thuringiensis 570 - Biología::577 - Ecología 570 - Biología::576 - Genética y evolución 570 - Biología::572 - Bioquímica Maíz - Enfermedades y plagas Spodoptera frugiperda Arsenophonus Biotipo maíz Endotoxinas BT Microbiota intestinal Resistencia Corn Strain Bt toxins Gut microbiota Resistance |
| title_short |
Evaluación de la microbiota del tracto gastrointestinal del biotipo de maíz de Spodoptera frugiperda en presencia de endotoxinas del Bacillus thuringiensis |
| title_full |
Evaluación de la microbiota del tracto gastrointestinal del biotipo de maíz de Spodoptera frugiperda en presencia de endotoxinas del Bacillus thuringiensis |
| title_fullStr |
Evaluación de la microbiota del tracto gastrointestinal del biotipo de maíz de Spodoptera frugiperda en presencia de endotoxinas del Bacillus thuringiensis |
| title_full_unstemmed |
Evaluación de la microbiota del tracto gastrointestinal del biotipo de maíz de Spodoptera frugiperda en presencia de endotoxinas del Bacillus thuringiensis |
| title_sort |
Evaluación de la microbiota del tracto gastrointestinal del biotipo de maíz de Spodoptera frugiperda en presencia de endotoxinas del Bacillus thuringiensis |
| dc.creator.fl_str_mv |
Castañeda Molina, Yuliana del Pilar |
| dc.contributor.advisor.none.fl_str_mv |
Cadavid Restrepo, Gloria Ester Saldamando Benjumea, Clara Inés |
| dc.contributor.author.none.fl_str_mv |
Castañeda Molina, Yuliana del Pilar |
| dc.contributor.educationalvalidator.none.fl_str_mv |
Moreno Herrera, Claudia Ximena |
| dc.contributor.researchgroup.spa.fl_str_mv |
Microbiodiversidad y Bioprospección |
| dc.subject.ddc.spa.fl_str_mv |
570 - Biología::577 - Ecología 570 - Biología::576 - Genética y evolución 570 - Biología::572 - Bioquímica |
| topic |
570 - Biología::577 - Ecología 570 - Biología::576 - Genética y evolución 570 - Biología::572 - Bioquímica Maíz - Enfermedades y plagas Spodoptera frugiperda Arsenophonus Biotipo maíz Endotoxinas BT Microbiota intestinal Resistencia Corn Strain Bt toxins Gut microbiota Resistance |
| dc.subject.lemb.none.fl_str_mv |
Maíz - Enfermedades y plagas |
| dc.subject.proposal.none.fl_str_mv |
Spodoptera frugiperda Arsenophonus |
| dc.subject.proposal.spa.fl_str_mv |
Biotipo maíz Endotoxinas BT Microbiota intestinal Resistencia |
| dc.subject.proposal.eng.fl_str_mv |
Corn Strain Bt toxins Gut microbiota Resistance |
| description |
ilustraciones, diagramas, tablas |
| publishDate |
2021 |
| dc.date.issued.none.fl_str_mv |
2021 |
| dc.date.accessioned.none.fl_str_mv |
2022-08-26T22:04:40Z |
| dc.date.available.none.fl_str_mv |
2022-08-26T22:04:40Z |
| dc.type.spa.fl_str_mv |
Trabajo de grado - Maestría |
| dc.type.driver.spa.fl_str_mv |
info:eu-repo/semantics/masterThesis |
| dc.type.version.spa.fl_str_mv |
info:eu-repo/semantics/acceptedVersion |
| dc.type.content.spa.fl_str_mv |
Text |
| dc.type.redcol.spa.fl_str_mv |
http://purl.org/redcol/resource_type/TM |
| status_str |
acceptedVersion |
| dc.identifier.uri.none.fl_str_mv |
https://repositorio.unal.edu.co/handle/unal/82152 |
| dc.identifier.instname.spa.fl_str_mv |
Universidad Nacional de Colombia |
| dc.identifier.reponame.spa.fl_str_mv |
Repositorio Institucional Universidad Nacional de Colombia |
| dc.identifier.repourl.spa.fl_str_mv |
https://repositorio.unal.edu.co/ |
| url |
https://repositorio.unal.edu.co/handle/unal/82152 https://repositorio.unal.edu.co/ |
| identifier_str_mv |
Universidad Nacional de Colombia Repositorio Institucional Universidad Nacional de Colombia |
| dc.language.iso.spa.fl_str_mv |
spa |
| language |
spa |
| dc.relation.references.spa.fl_str_mv |
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Paddock, K.J., Pereira, A.E., Finke, D.L., Ericsson, A.C., Hibbard, B.E. and Shelby, K.S. (2021), Host resistance to Bacillus thuringiensis is linked to altered bacterial community within a specialist insect herbivore. Mol Ecol, 30: 5438-5453. https://doi.org/10.1111/mec.15875 Pardo-Lopez, L., Soberon, M., & Bravo, A. (2013). Bacillus thuringiensis insecticidal three-domain Cry toxins: mode of action, insect resistance and consequences for crop protection. FEMS microbiology reviews, 37(1), 3-22. Patil, C. D., Borase, H. P., Salunke, B. K., & Patil, S. V. (2013). Alteration in Bacillus thuringiensis toxicity by curing gut flora: novel approach for mosquito resistance management. Parasitology research, 112(9), 3283-3288. Paul J. McMurdie phyloseq: An R package for reproducible interactive analysis and graphics of microbiome censos data., 2013, R package version 1.19 Peterson, B., Bezuidenhout, C. C., & Van den Berg, J. (2017). 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Wang, Q.; Ren, M.; Liu, X.; Xia, H.; Chen, K. Peptidoglycan recognition proteins in insect immunity.Mol. Immunol.2019,106, 69–76 Yamamoto, S., & Harayama, S. (1995). PCR amplification and direct sequencing of gyrB genes with universal primers and their application to the detection and taxonomic analysis of Pseudomonas putida strains. Applied and environmental microbiology, 61(3), 1104-1109. Zhu, Y. C., Kramer, K. J., Oppert, B., & Dowdy, A. K. (2000). cDNAs of aminopeptidase-like protein genes from Plodia interpunctella strains with different susceptibilities to Bacillus thuringiensis toxins. Insect Biochemistry and Molecular Biology, 30(3), 215-224 |
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Reconocimiento 4.0 Internacionalhttp://creativecommons.org/licenses/by/4.0/info:eu-repo/semantics/openAccesshttp://purl.org/coar/access_right/c_abf2Cadavid Restrepo, Gloria Ester678bdd51eb1e2c1316a16d5074303830600Saldamando Benjumea, Clara Inéseb84ab30edba9019a94e8184c42a2d41600Castañeda Molina, Yuliana del Pilar835da70c24b3c123d000494a5a241de9Moreno Herrera, Claudia XimenaMicrobiodiversidad y Bioprospección2022-08-26T22:04:40Z2022-08-26T22:04:40Z2021https://repositorio.unal.edu.co/handle/unal/82152Universidad Nacional de ColombiaRepositorio Institucional Universidad Nacional de Colombiahttps://repositorio.unal.edu.co/ilustraciones, diagramas, tablasSpodoptera frugiperda (Lepidoptera, Noctuidae), es reconocida como una plaga polífaga primaria de cultivos de maíz (Zea mays) y arroz (Oryza sativa) en todo el continente americano y recientemente en África, Asia y Australia. En Colombia, esta polilla ha divergido en dos biotipos morfológicamente idénticos en estado de larva, pero morfológicamente diferentes en estado adulto, los cuales se han denominado con base en la asociación al alimento que consumen con mayor preferencia: biotipos de maíz (asociado a los cultivos del maíz, algodón, sorgo y caña de azúcar) y biotipo arroz (asociado a arroz y pasto). Desde hace varias décadas, se ha descrito que en los insectos, las capacidades de defensa, digestión, captación de nutrientes, incluso la degradación de compuestos tóxicos como insecticidas y endotoxinas está fuertemente influenciada por su microbiota intestinal. Se ha encontrado que en Colombia, el biotipo maíz ha desarrollado resistencia a las endotoxinas de Bacillus thuringiensis (Bt) Cry 1Ac y Cry 1Ab, por lo que la microbiota intestinal podría tener un efecto modulador en la toxicidad de las mismas. En este estudio, se llevó a cabo la caracterización de la microbiota intestinal de larvas de S. frugiperda biotipo maíz en presencia/ausencia de endotoxinas de Bt y de antibióticos para determinar el papel de la microbiota en la respuesta del insecto al Bt. Para ello se implementó el uso de métodos cultivo dependientes y cultivo independientes mediante análisis de secuenciación Sanger y NGS (Next Generation Sequence). Adicionalmente se determinó la presencia de microorganismos endosimbiontes con potencial en control biológico como Wolbachia, Arsenophonus, Microsporidia, Spiroplasma, y Cardinium, usando cebadores específicos. Los aislados bacterianos intestinales fueron identificados a partir de caracterización molecular, mediante la secuenciación de los genes RNAr 16S y girasa, previo análisis de la región intergénica ribosomal (ITS) y adicionalmente la caracterización macro y microscópica de las colonias fue realizada. Los resultados indicaron que los 15 aislamientos bacterianos pertenecen a las especies Enterococcus mundtii, Enterococcus sileciacus, Enterococcus gallinarum y Enterococcus casseliflavus. Los resultados de NGS (Illumina Miseq) reportaron que, Firmicutes y Proteobacteria fueron los filos más abundantes en las muestras; un total de 790.011 lecturas fueron analizadas y asignadas por similaridad a 2439 ASVs (Del inglés Amplicon Sequence Variant). Con relación a la presencia de endosimbiontes, se encontró una prevalencia del 100% de Arsenophonus, por métodos de PCR convencional y NGS, en todas las muestras evaluadas; sin embargo, otros simbiontes no fueron detectados en las condiciones evaluadas. En este estudio se reportó la prevalencia del género Enterococcus así como ya se ha descrito en otros estudios anteriores realizados en el mismo biotipo. Adicionalmente se encontró que posterior al ensayo con antibióticos, la diversidad de la microbiota intestinal aumentó, lo cual podría sugerir la actuación de los antibióticos sobre bacterias del género Enterococcus lo que consecuentemente permitió un aumento en la diversidad bacteriana ya que se eliminó el género más competitivo y abundante del tracto intestinal del insecto. Por otro lado, se lograron identificar los géneros Burkholderia e Ileibacterium en aquellos tratamientos con presencia de Bt, sugiriendo que estas bacterias podrían realzar la respuesta del insecto a la endotoxina. Finalmente, este es, el primer reporte del género Arsenophonus en S. frugiperda, este endosimbionte es importante en insectos ya que ha sido relacionado con la respuesta de tolerancia a insecticidas que han sido utilizados para su control. Este trabajo es una primera aproximación al conocimiento de la microbiota de S. frugiperda biotipo maíz y su dinámica en presencia de endotoxinas lo que puede ayudar a comprender su papel potencial en la respuesta del insecto al control por Bt. (Texto tomado de la fuente)Spodoptera frugiperda (Lepidoptera, Noctuidae), is recognized as a polyphagous pest and primary pest of corn (Zea mays) and rice (Oryza sativa) in America and recently in Africa, Asia and Australia. In Colombia, this moth has diverged in two strains that are morphologically identical at the larvae instar but are morphologically different at adult stages. These strains have been named according to the host they preferentially consume that are the corn strain (associated to corn, cotton, sorghum and sugar cane) and the rice strain (associated to rice and pasture grasses). Decades ago, it has been described that the capacity of insects’ defense, digestion, nutrients absorption and even degradation of toxic compounds such as insecticides and endotoxins are influenced by their microbiota. It has been found that in Colombia, the corn strain has developed resistance to Bacillus thuringiensis (Bt) endotoxins Cry1Ac and Cry1Ab and gut microbiota might have a modulatory effect on the response of this insect towards these endotoxins. In this study, the characterization of the gut microbiota of S. frugiperda larvae was carried out un presence/absence of Bt endotoxins and antibiotics to determine the role that microbiota plays in the response of the insect to Bt. For that, the use of cultivate dependent and independent methods were employed by using Sanger and NGS (Next Generation Sequence) sequencing. Additionally, detection of endosymbionts was also done to identify Wolbachia, Arsenophonus, Microsporidia, Spiroplasma, and Cardinium.by using specific primers for them. Bacteria isolates obtanied from the gut were identified based on a molecular characterization by sequencing the genes RNAr 16S, girase and ITS. Also, a macroscopic and microscopic characterization of the colonies was done allowing the identification of 15 different isolates that belong to the species Enterococcus mundtii, Enterococcus sileciacus, Enterococcus gallinarum y Enterococcus casseliflavus. NGS (Illumina Miseq) results reported the presence of the phyla Firmicutes and Proteobacteria as the most abundant groups; a total of 790.011 reads grouped into 2439 ASVs according to their genetic similarities. The presence of endosymbionts analysis showed a prevalence of the genus Arsenophonus in all samples based on conventional PCR and NGS; however, other endosymbionts were not detected. In this study, the prevalence of the genus Enterococcus was demonstrated, same results have been shown in other studies made on the species. Additionally, after the expose of gut microbiota to antibiotics an increment of bacteria diversity was observed, suggesting that these antibiotics eliminated this abundant genus found in S. frugiperda gut, and thus they eliminated the most competitive bacteria genus detected in this insect. On the other hand, the genus Burkholderia and Ileibacterium were mainly found in the bioassay of bacteria and presence of Bt, meaning that these bacteria might enhance the response of this insect to the endotoxin. Finally, this is the first report of the genus Arsenophonus in S. frugiperda and this endosymbiont is important in insects since it is related to the tolerance response of insects to insecticides that have been used for pest control. This work is a first approximation to the knowledge of S. frugiperda gut bacteria corn strain and its dynamics in the presence of endotoxins, which can help to understand its potential role in the response of the insect to control by Bt.MaestríaMagíster en Ciencias - BiotecnologíaEcología MicrobianaÁrea Curricular Biotecnologíaxx, 142 páginasapplication/pdfspaUniversidad Nacional de ColombiaMedellín - Ciencias - Maestría en Ciencias - BiotecnologíaEscuela de biocienciasFacultad de CienciasMedellín, ColombiaUniversidad Nacional de Colombia - Sede Medellín570 - Biología::577 - Ecología570 - Biología::576 - Genética y evolución570 - Biología::572 - BioquímicaMaíz - Enfermedades y plagasSpodoptera frugiperdaArsenophonusBiotipo maízEndotoxinas BTMicrobiota intestinalResistenciaCorn StrainBt toxinsGut microbiotaResistanceEvaluación de la microbiota del tracto gastrointestinal del biotipo de maíz de Spodoptera frugiperda en presencia de endotoxinas del Bacillus thuringiensisEvaluation of Spodoptera frugiperda (corn strain) gut microbiota in presence of Bacillus thuringiensis endotoxinsTrabajo de grado - Maestríainfo:eu-repo/semantics/masterThesisinfo:eu-repo/semantics/acceptedVersionTexthttp://purl.org/redcol/resource_type/TMAdamczyk, John J, Jonathan W Holloway, Billy R Leonard, and Jerry B Graves. 1997. “Susceptibility of Fall Armyworm Collected from Different Plant Hosts to Selected.” 28(August): 21–28.Abdelhadi AA, Elarabi NI, Salim RG, Sharaf AN, Abosereh NA (2016) Identification, characterization and genetic improvement of bacteriocin producing lactic acid bacteria. 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Insect Biochemistry and Molecular Biology, 30(3), 215-224Código QUIPU: 2020100134MinCienciasEstudiantesInvestigadoresMaestrosLICENSElicense.txtlicense.txttext/plain; charset=utf-81748https://repositorio.unal.edu.co/bitstream/unal/82152/1/license.txt8a4605be74aa9ea9d79846c1fba20a33MD51ORIGINAL1033687052_2021.pdf1033687052_2021.pdfTesis de Maestría en Ciencias - Biotecnologíaapplication/pdf4431307https://repositorio.unal.edu.co/bitstream/unal/82152/2/1033687052_2021.pdf30d6ba6cf42a413083ab92cd305fa29eMD52THUMBNAIL1033687052_2021.pdf.jpg1033687052_2021.pdf.jpgGenerated Thumbnailimage/jpeg5526https://repositorio.unal.edu.co/bitstream/unal/82152/3/1033687052_2021.pdf.jpg8e8801aec65532f2ffdd65a734db2b52MD53unal/82152oai:repositorio.unal.edu.co:unal/821522023-08-08 23:04:02.72Repositorio Institucional Universidad Nacional de Colombiarepositorio_nal@unal.edu.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 |