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Variabilidade entre isolados de Cowpea severe mosaic virus por meio de caracterização biológica, sorológica e molecular

Detalhes bibliográficos
Ano de defesa: 2017
Autor(a) principal: Silva, Ana Kelly Firmino da
Orientador(a): Aragão, Fernando Antonio Souza de
Banca de defesa: Não Informado pela instituição
Tipo de documento: Tese
Tipo de acesso: Acesso aberto
Idioma: por
Instituição de defesa: Não Informado pela instituição
Programa de Pós-Graduação: Não Informado pela instituição
Departamento: Não Informado pela instituição
País: Não Informado pela instituição
Palavras-chave em Português:
Link de acesso: http://www.repositorio.ufc.br/handle/riufc/25569
Resumo: Cowpea (Vigna unguiculata subsp. unguiculata) is grown as staple food for the rural and urban populations inf the North and Northeast Brazil. It has great importance as a generator and income, being cultivated by small to medium producers. Brazil is the largest producer of beans [ commob bean (Phaseolus vulgaris) and cowpea] with production of 2.5 milion tons in the 2015/16 harvest, being the States of Paraná and Minas Gerais the largest producers of common bean, and State of Ceará in ninth position. Several factors affect the cowpea crop influencing the quality and quantity produced, highlighting the viruses as responsible for large losses in the crop. Several species of virus can naturally infect cowpea, especially the Cowpea severe mosaic virus (CPSMV). The objective of this research was to study the genetic variability among eight isolates of CPSMV obtained in different States of the Northeast region, through biological, serological and molecular characterization studies. The experiments were carried out in a greenhouse and in the Plant Virology Laboratory of the Federal University of Ceará. The CPSMV-MC isolate was purified from cowpea cv. Macaibo, obtaining a virus preparation with a concentration of 64.83 mg of virus per kg of infected tissue and antiserum title of 1: 1024 in Double Immune Diffusion Test. The biological study, by partial range of host plants showed symptomatological difference between the CPSMV-MC isolate of the others. The results of the serological study showed that the CPSMV-MC isolate may be serologically distinct when using antiserum produced from this isolate. The electrophoretic analyzes revealed two bands related to the weight of the capsidial protein formed by 22.5 kDa and 39 kDa structural proteins. Comparison between variations in the PTA-ELISA technique, showed that IP-PTA-ELISA, was efficient and reliable for detection of CPSMV suppressing the problems of virus of the genus Comovirus that do not adhere adequately to the Of ELISA plates. In the molecular study, the primers designed for the partial genome of RNA1 and RNA2 regions could be used for both detection and sequencing of the virus. The phylogenetic analyzes of the sequences of the CPSMV isolates, using primers for the RNA1 region revealed a difference between the CPSMV-MC isolate and the others, making it possible to fit them into two groups. One group that causes severe symptoms, adding seven isolates of CPSMV and the other group with only one isolate that causes mild symptom (CPSMV-MC) in cowpea. The study of the interaction between the virus that infects cowpea 'Setentão' showed strong synergism between CPSMV, Cowpea aphid-borne mosaic virus (CABMV) and Cucumber mosaic virus (CMV), leading in some cases to death of the plant, mainly when interacting Of these three viruses, and two of these isolates showed severe symptoms in the plants when in simple infection. Studies of this nature involving aspects related to pathogens and the plant produce important information in the research programs that aim at the selection and production of virus resistant cultivars.
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spelling Silva, Ana Kelly Firmino daAragão, Fernando Antonio Souza de2017-09-06T20:35:43Z2017-09-06T20:35:43Z2017SILVA, Ana Kelly Firmino. Variabilidade entre isolados de Cowpea severe mosaic virus por meio de caracterização biológica, sorológica e molecular. 2017. 81 f. Tese (Doutorado em Agronomia/Fitotecnia)-Universidade Federal do Ceará, Fortaleza, 2017.http://www.repositorio.ufc.br/handle/riufc/25569Cowpea (Vigna unguiculata subsp. unguiculata) is grown as staple food for the rural and urban populations inf the North and Northeast Brazil. It has great importance as a generator and income, being cultivated by small to medium producers. Brazil is the largest producer of beans [ commob bean (Phaseolus vulgaris) and cowpea] with production of 2.5 milion tons in the 2015/16 harvest, being the States of Paraná and Minas Gerais the largest producers of common bean, and State of Ceará in ninth position. Several factors affect the cowpea crop influencing the quality and quantity produced, highlighting the viruses as responsible for large losses in the crop. Several species of virus can naturally infect cowpea, especially the Cowpea severe mosaic virus (CPSMV). The objective of this research was to study the genetic variability among eight isolates of CPSMV obtained in different States of the Northeast region, through biological, serological and molecular characterization studies. The experiments were carried out in a greenhouse and in the Plant Virology Laboratory of the Federal University of Ceará. The CPSMV-MC isolate was purified from cowpea cv. Macaibo, obtaining a virus preparation with a concentration of 64.83 mg of virus per kg of infected tissue and antiserum title of 1: 1024 in Double Immune Diffusion Test. The biological study, by partial range of host plants showed symptomatological difference between the CPSMV-MC isolate of the others. The results of the serological study showed that the CPSMV-MC isolate may be serologically distinct when using antiserum produced from this isolate. The electrophoretic analyzes revealed two bands related to the weight of the capsidial protein formed by 22.5 kDa and 39 kDa structural proteins. Comparison between variations in the PTA-ELISA technique, showed that IP-PTA-ELISA, was efficient and reliable for detection of CPSMV suppressing the problems of virus of the genus Comovirus that do not adhere adequately to the Of ELISA plates. In the molecular study, the primers designed for the partial genome of RNA1 and RNA2 regions could be used for both detection and sequencing of the virus. The phylogenetic analyzes of the sequences of the CPSMV isolates, using primers for the RNA1 region revealed a difference between the CPSMV-MC isolate and the others, making it possible to fit them into two groups. One group that causes severe symptoms, adding seven isolates of CPSMV and the other group with only one isolate that causes mild symptom (CPSMV-MC) in cowpea. The study of the interaction between the virus that infects cowpea 'Setentão' showed strong synergism between CPSMV, Cowpea aphid-borne mosaic virus (CABMV) and Cucumber mosaic virus (CMV), leading in some cases to death of the plant, mainly when interacting Of these three viruses, and two of these isolates showed severe symptoms in the plants when in simple infection. Studies of this nature involving aspects related to pathogens and the plant produce important information in the research programs that aim at the selection and production of virus resistant cultivars.O feijão caupi Vigna unguiculata subsp. unguiculata constitui um alimento básico das populações rurais e urbanas das regiões Norte e Nordeste, possui grande importância como gerador de emprego e renda, sendo cultivado por pequenos e médios produtores. O Brasil é o maior produtor de feijão [feijão comum (Phaseolus vulgaris) e feijão caupi] com produção de 2,5 mil toneladas na safra de 2015/16, sendo os estados do Paraná e Minas Gerais os maiores produtores de feijão comum, ficando o estado do Ceará na nona posição. Vários fatores afetam a cultura do feijão caupi influenciando na qualidade e na quantidade produzida, destacando as viroses como responsáveis por grandes perdas na cultura. Várias espécies de vírus podem infectar naturalmente o feijão caupi, destacando-se o Cowpea severe mosaic virus (CPSMV). O objetivo desta pesquisa foi estudar a variabilidade genética entre oito isolados de CPSMV obtidos em diferentes Estados da Região Nordeste, mediante estudos de caracterização biológica, sorológica e molecular. Os experimentos realizados foram conduzidos em casa de vegetação e no Laboratório de Virologia Vegetal da Universidade Federal do Ceará. O isolado de CPSMV-MC foi purificado a partir de plantas de feijão caupi cv. Macaibo, obtendo uma preparação viral com concentração de 64,83 mg de vírus por Kg de tecido infectado e título do antissoro de 1:1.024 em teste de Dupla Difusão em Ágar (DDA). No estudo biológico, por gama parcial de plantas hospedeiras mostrou diferença sintomatológica entre o isolado CPSMV-MC dos demais. O resultado do estudo sorológico mostrou que o isolado CPSMV-MC pode ser sorologicamente distinto quando utilizando antissoro produzido a partir deste isolado. As análises eletroforética revelaram bandas referentes ao peso da proteína capsidial formada por duas proteínas estruturais de 22,5 kDa e de 39 kDa, aproximadamente. Estudo de comparação entre PTA-ELISA e IP-PTA-ELISA, variações da técnica de ELISA, mostrou que o IP-ELISA foi eficiente e confiável para a detecção do CPSMV contornando alguns problemas de vírus do gênero Comovirus que não aderem adequadamente aos fundos dos orifícios das placas de ELISA. No estudo molecular, os primers desenhados para o genoma parcial de regiões dos RNA1 e RNA2 podem ser utilizados tanto para detecção quanto para sequenciamento do vírus. As análises filogenéticas das sequências dos isolados de CPSMV utilizando primers para a região do RNA1 revelaram diferença entre o isolado CPSMV-MC e os demais, possibilitando o enquadramento dos mesmos em dois grupos. Um grupo que causam sintomas severos, agregando sete isolados de CPSMV e o outro grupo com apenas um isolado que causa sintoma leve (CPSMV-MC) em feijão caupi. O estudo de interação entre vírus que infecta o feijão caupi ‘Setentão’ mostrou forte sinergismo entre CPSMV, Cowpea aphid-borne mosaic virus (CABMV) e Cucumber mosaic virus (CMV), levando em alguns caso morte da planta, principalmente, quando da interação desses três vírus, sendo que dois desse isolados apresentaram sintomas severos nas plantas quando em infecção simples. Estudos desta natureza, envolvendo aspectos relacionados ao patógenos e a planta produzem informações importantes nos programas de pesquisam que visam a seleção e produção de cultivares resistentes a vírus.Feijão caupiVigna unguiculataVariabilidade entre isolados de Cowpea severe mosaic virus por meio de caracterização biológica, sorológica e molecularVariability between isolates of Cowpea severe mosaic virus of biological, serological and molecular characterizationinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisporreponame:Repositório Institucional da Universidade Federal do Ceará (UFC)instname:Universidade Federal do Ceará (UFC)instacron:UFCinfo:eu-repo/semantics/openAccessORIGINAL2017_tese_akfsilva.pdf2017_tese_akfsilva.pdfapplication/pdf1731430http://repositorio.ufc.br/bitstream/riufc/25569/3/2017_tese_akfsilva.pdf52fe9ff98614a1fa8e493ba2d12f68efMD53LICENSElicense.txtlicense.txttext/plain; charset=utf-81748http://repositorio.ufc.br/bitstream/riufc/25569/2/license.txt8a4605be74aa9ea9d79846c1fba20a33MD52riufc/255692022-09-05 11:16:18.99oai:repositorio.ufc.br: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Repositório InstitucionalPUBhttp://www.repositorio.ufc.br/ri-oai/requestbu@ufc.br || repositorio@ufc.bropendoar:2022-09-05T14:16:18Repositório Institucional da Universidade Federal do Ceará (UFC) - Universidade Federal do Ceará (UFC)false
dc.title.pt_BR.fl_str_mv Variabilidade entre isolados de Cowpea severe mosaic virus por meio de caracterização biológica, sorológica e molecular
dc.title.en.pt_BR.fl_str_mv Variability between isolates of Cowpea severe mosaic virus of biological, serological and molecular characterization
title Variabilidade entre isolados de Cowpea severe mosaic virus por meio de caracterização biológica, sorológica e molecular
spellingShingle Variabilidade entre isolados de Cowpea severe mosaic virus por meio de caracterização biológica, sorológica e molecular
Silva, Ana Kelly Firmino da
Feijão caupi
Vigna unguiculata
title_short Variabilidade entre isolados de Cowpea severe mosaic virus por meio de caracterização biológica, sorológica e molecular
title_full Variabilidade entre isolados de Cowpea severe mosaic virus por meio de caracterização biológica, sorológica e molecular
title_fullStr Variabilidade entre isolados de Cowpea severe mosaic virus por meio de caracterização biológica, sorológica e molecular
title_full_unstemmed Variabilidade entre isolados de Cowpea severe mosaic virus por meio de caracterização biológica, sorológica e molecular
title_sort Variabilidade entre isolados de Cowpea severe mosaic virus por meio de caracterização biológica, sorológica e molecular
author Silva, Ana Kelly Firmino da
author_facet Silva, Ana Kelly Firmino da
author_role author
dc.contributor.author.fl_str_mv Silva, Ana Kelly Firmino da
dc.contributor.advisor1.fl_str_mv Aragão, Fernando Antonio Souza de
contributor_str_mv Aragão, Fernando Antonio Souza de
dc.subject.por.fl_str_mv Feijão caupi
Vigna unguiculata
topic Feijão caupi
Vigna unguiculata
description Cowpea (Vigna unguiculata subsp. unguiculata) is grown as staple food for the rural and urban populations inf the North and Northeast Brazil. It has great importance as a generator and income, being cultivated by small to medium producers. Brazil is the largest producer of beans [ commob bean (Phaseolus vulgaris) and cowpea] with production of 2.5 milion tons in the 2015/16 harvest, being the States of Paraná and Minas Gerais the largest producers of common bean, and State of Ceará in ninth position. Several factors affect the cowpea crop influencing the quality and quantity produced, highlighting the viruses as responsible for large losses in the crop. Several species of virus can naturally infect cowpea, especially the Cowpea severe mosaic virus (CPSMV). The objective of this research was to study the genetic variability among eight isolates of CPSMV obtained in different States of the Northeast region, through biological, serological and molecular characterization studies. The experiments were carried out in a greenhouse and in the Plant Virology Laboratory of the Federal University of Ceará. The CPSMV-MC isolate was purified from cowpea cv. Macaibo, obtaining a virus preparation with a concentration of 64.83 mg of virus per kg of infected tissue and antiserum title of 1: 1024 in Double Immune Diffusion Test. The biological study, by partial range of host plants showed symptomatological difference between the CPSMV-MC isolate of the others. The results of the serological study showed that the CPSMV-MC isolate may be serologically distinct when using antiserum produced from this isolate. The electrophoretic analyzes revealed two bands related to the weight of the capsidial protein formed by 22.5 kDa and 39 kDa structural proteins. Comparison between variations in the PTA-ELISA technique, showed that IP-PTA-ELISA, was efficient and reliable for detection of CPSMV suppressing the problems of virus of the genus Comovirus that do not adhere adequately to the Of ELISA plates. In the molecular study, the primers designed for the partial genome of RNA1 and RNA2 regions could be used for both detection and sequencing of the virus. The phylogenetic analyzes of the sequences of the CPSMV isolates, using primers for the RNA1 region revealed a difference between the CPSMV-MC isolate and the others, making it possible to fit them into two groups. One group that causes severe symptoms, adding seven isolates of CPSMV and the other group with only one isolate that causes mild symptom (CPSMV-MC) in cowpea. The study of the interaction between the virus that infects cowpea 'Setentão' showed strong synergism between CPSMV, Cowpea aphid-borne mosaic virus (CABMV) and Cucumber mosaic virus (CMV), leading in some cases to death of the plant, mainly when interacting Of these three viruses, and two of these isolates showed severe symptoms in the plants when in simple infection. Studies of this nature involving aspects related to pathogens and the plant produce important information in the research programs that aim at the selection and production of virus resistant cultivars.
publishDate 2017
dc.date.accessioned.fl_str_mv 2017-09-06T20:35:43Z
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dc.identifier.citation.fl_str_mv SILVA, Ana Kelly Firmino. Variabilidade entre isolados de Cowpea severe mosaic virus por meio de caracterização biológica, sorológica e molecular. 2017. 81 f. Tese (Doutorado em Agronomia/Fitotecnia)-Universidade Federal do Ceará, Fortaleza, 2017.
dc.identifier.uri.fl_str_mv http://www.repositorio.ufc.br/handle/riufc/25569
identifier_str_mv SILVA, Ana Kelly Firmino. Variabilidade entre isolados de Cowpea severe mosaic virus por meio de caracterização biológica, sorológica e molecular. 2017. 81 f. Tese (Doutorado em Agronomia/Fitotecnia)-Universidade Federal do Ceará, Fortaleza, 2017.
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