Estudo de isolados brasileiros de potyvírus causadores de endurecimento dos frutos do maracujazeiro e análise da resistência à doença em linhagens transgênicas R1 de maracujá-amarelo
Autor(a) principal: | |
---|---|
Data de Publicação: | 2006 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | LOCUS Repositório Institucional da UFV |
Texto Completo: | http://locus.ufv.br/handle/123456789/1040 |
Resumo: | Passionfruit woodiness is the most important disease of passion flower. The disease can be caused by the potyviruses Passionfruit woodiness virus (PWV), Cowpea aphid-borne mosaic virus (CABMV) and East Asian Passiflora virus (EAPV). Infected plants display mosaic and woodiness of the fruit. In Brazil, the disease occurs in all states where passion flower is grown, and the causeal agent has always been identified as PWV based on biological and serological properties. Management strategies based on vector control with insecticides, cross-protection and natural resistance are inneficient. The objectives of this work were to characterize at the molecular level a number of Brazilian isolates of potyvirus causing passionfruit woodiness and to evaluate the resistance of transgenic yellow passion flower (Passiflora edulis f. flavicarpa) plants expressing a genomic fragment from a woodinessinducing potyvirus isolate from Brazil. Foliar samples of yellow passion flower showing typical symptoms of the disease were collected collected in the states of Bahia, Espírito Santo, Minas Gerais, Paraíba, Pernambuco and Sergipe. The presence of the virus was confirmed by serology and host range. The viral isolates were capable of infecting plants of common bean and cowpea, although differing in the intensity of symptoms induced in these hosts. Serological relationship between the potyvirus isolates from passion flower Cowpea aphidborne mosaic virus (CABMV) was demonstrated by indirect ELISA. The sequence of the capsid protein coding region was determined for all isolates. Sequence comparisons with other potyviruses indicated a level of identify from 85 to 94% with CABMV isolates. Identity with PWV isolate K ranged from 54 to 70%. Phylogenetic analysis based on the deduced capsid protein amino acid sequences clustered the Brazilian isolates together with CABMV isolates, distant from PWV isolates. Together, these results indicated that the potyvirus isolates from passion flower represent a strain of CABMV. In an attempt to effectively control passionfruit woodiness, transgenic yellow passionfruit plants (Passiflora edulis f. flavicarpa) were generated which express an untranslatable RNA corresponding to the replicase and capsid protein coding regions of a CABMV isolate from Minas Gerais (MGAvr). One R0 plant (TE5-10) was resistant to the MG-Avr isolate due to RNA silencing. However, this plant was susceptible to a second isolate of the virus (PE-Bnt). In order to verify the effect of gene dosage in the spectrum of resistance, several crosses were carried out between the TE5-10 plant, a susceptible R0 plant (T2-5) ans a non-transformed plant. Also, self-pollination of the TE5-10 plant was carried out in orde to generate transgene-homozigous R1 plants. The plants obtained from these crosses were challenged with three CABMV isolates (MG-Avr, PE-Bnt and SE-Nps). One R1 plant obtained from self-pollination (named TE5-10-15J) was resistant to the three isolates. This plant was challenged with an additional four CABMV isolates (PB-Alh, BA-Itb, ES-Vni and PA-Iga), being resistant to all of them based on symptom development. Northern analysis demonstrated the lack of transgenic mRNA accumulation in these plants, indicating that resistance is due to RNA silencing. |
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Nascimento, Ana Verônica Silva dohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4760240Y3Otoni, Wagner Camposhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4786133Y6Zerbini Júnior, Francisco Murilohttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783743U5Pio-ribeiro, Gilvanhttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4793337P7Zerbini, Poliane Alfenashttp://lattes.cnpq.br/8632328159533071Oliveira, Rosângela D arc de Limahttp://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4788026Z32015-03-26T12:41:57Z2007-02-152015-03-26T12:41:57Z2006-08-31NASCIMENTO, Ana Verônica Silva do. Study of Brazilian potyvirus isolates causing passionfruit woodiness and analysis of resistance to the disease in R1 transgenic lines of passionflower. 2006. 55 f. Tese (Doutorado em Etiologia; Epidemiologia; Controle) - Universidade Federal de Viçosa, Viçosa, 2006.http://locus.ufv.br/handle/123456789/1040Passionfruit woodiness is the most important disease of passion flower. The disease can be caused by the potyviruses Passionfruit woodiness virus (PWV), Cowpea aphid-borne mosaic virus (CABMV) and East Asian Passiflora virus (EAPV). Infected plants display mosaic and woodiness of the fruit. In Brazil, the disease occurs in all states where passion flower is grown, and the causeal agent has always been identified as PWV based on biological and serological properties. Management strategies based on vector control with insecticides, cross-protection and natural resistance are inneficient. The objectives of this work were to characterize at the molecular level a number of Brazilian isolates of potyvirus causing passionfruit woodiness and to evaluate the resistance of transgenic yellow passion flower (Passiflora edulis f. flavicarpa) plants expressing a genomic fragment from a woodinessinducing potyvirus isolate from Brazil. Foliar samples of yellow passion flower showing typical symptoms of the disease were collected collected in the states of Bahia, Espírito Santo, Minas Gerais, Paraíba, Pernambuco and Sergipe. The presence of the virus was confirmed by serology and host range. The viral isolates were capable of infecting plants of common bean and cowpea, although differing in the intensity of symptoms induced in these hosts. Serological relationship between the potyvirus isolates from passion flower Cowpea aphidborne mosaic virus (CABMV) was demonstrated by indirect ELISA. The sequence of the capsid protein coding region was determined for all isolates. Sequence comparisons with other potyviruses indicated a level of identify from 85 to 94% with CABMV isolates. Identity with PWV isolate K ranged from 54 to 70%. Phylogenetic analysis based on the deduced capsid protein amino acid sequences clustered the Brazilian isolates together with CABMV isolates, distant from PWV isolates. Together, these results indicated that the potyvirus isolates from passion flower represent a strain of CABMV. In an attempt to effectively control passionfruit woodiness, transgenic yellow passionfruit plants (Passiflora edulis f. flavicarpa) were generated which express an untranslatable RNA corresponding to the replicase and capsid protein coding regions of a CABMV isolate from Minas Gerais (MGAvr). One R0 plant (TE5-10) was resistant to the MG-Avr isolate due to RNA silencing. However, this plant was susceptible to a second isolate of the virus (PE-Bnt). In order to verify the effect of gene dosage in the spectrum of resistance, several crosses were carried out between the TE5-10 plant, a susceptible R0 plant (T2-5) ans a non-transformed plant. Also, self-pollination of the TE5-10 plant was carried out in orde to generate transgene-homozigous R1 plants. The plants obtained from these crosses were challenged with three CABMV isolates (MG-Avr, PE-Bnt and SE-Nps). One R1 plant obtained from self-pollination (named TE5-10-15J) was resistant to the three isolates. This plant was challenged with an additional four CABMV isolates (PB-Alh, BA-Itb, ES-Vni and PA-Iga), being resistant to all of them based on symptom development. Northern analysis demonstrated the lack of transgenic mRNA accumulation in these plants, indicating that resistance is due to RNA silencing.O endurecimento dos frutos é a doença mais importante da cultura do maracujazeiro. Essa doença pode ser causada pelos potyvírus Passionfruit woodiness virus (PWV), Cowpea aphid-borne mosaic virus (CABMV) e East Asian Passiflora Virus (EAPV). Plantas infectadas apresentam mosaico e frutos com endurecimento do pericarpo. No Brasil, a doença está presente nos principais estados produtores, e o agente causal sempre foi identificado como PWV com base em características biológicas e sorológicas. Estratégias de controle baseadas no controle do inseto vetor com inseticida, em proteção cruzada e em resistência natural tem se mostrado ineficientes. Os objetivos deste trabalho foram caracterizar em nível molecular isolados brasileiros de potyvírus causando endurecimento dos frutos do maracujazeiro e avaliar a resistência ao vírus em plantas transgênicas de maracujá-amarelo (Passiflora edulis f. flavicarpa) expressando um fragmento do genoma de um isolado brasileiro de potyvírus causador do endurecimento dos frutos. Amostras foliares de plantas de maracujá-amarelo com sintomas típicos de endurecimento dos frutos foram coletadas nos estados da Bahia, Espírito Santo, Minas Gerais, Paraíba, Pernambuco e Sergipe. A presença do vírus foi comprovada por meio de teste sorológico e gama de hospedeiros. Os isolados virais foram capazes de infectar plantas de feijoeiro e de feijão caupi, porém apresentando reações diferenciais quanto à intensidade dos sintomas induzidos nesses hospedeiros. ELISA indireto demonstrou que os isolados obtidos a partir de plantas de maracujá são sorologicamente relacionados entre si e com o potyvírus CABMV. As seqüências de aminoácidos da proteína capsidial (CP) foram determinadas para todos os isolados. A comparação dessas seqüências com as de outros potyvírus indicou identidade de 85 a 94% com isolados de CABMV. A identidade com o isolado K de PWV foi de 54 a 70%. Análise filogenética realizada a partir das seqüências de aminoácidos da CP, agrupou os isolados em estudo junto aos isolados de CABMV e distante de isolados de PWV. Em conjunto, os resultados indicam que os isolados de maracujá analisados constituem na verdade uma estirpe do CABMV. Na busca de uma estratégia eficiente de controle do endurecimento dos frutos, foram produzidas plantas transgênicas de maracujá-amarelo expressando um RNA nãotraduzível correspondente aos genes nib e cp de um isolado do CABMV de Minas Gerais (MG-Avr). Uma planta transgênica R0 (TE5-10) mostrou-se resistente ao isolado MG-Avr por meio de silenciamento gênico pós-transcricional, sendo porém suscetível a um isolado de Pernambuco (PE-Bnt). Para estudar o efeito da dosagem gênica no espectro da resistência foram realizados cruzamentos entre a planta resistente TE5-10, uma planta R0 suscetível (T2-5) e uma planta não-transformada. Além disso, foram realizadas autofecundações da planta TE5-10 com o objetivo de obter plantas com o transgene em homozigose. As plantas obtidas a partir desses cruzamentos foram inoculadas com três isolados do vírus (MG-Avr, PE-Bnt e SE-Nps). Uma planta R1 (TE5-10-15J) obtida a partir da autofecundação de planta R0 TE5-10 mostrou-se resistente aos três isolados. Essa planta foi desafiada com outros quatros isolados de CABMV (BA-Itb, PB-Alh, ES-Vni e PA-Iga), mostrando-se resistentes a todos eles. A análise do acúmulo de mRNA transgênico por meio de hibridização indicam que o mecanismo de silenciamento gênico é responsável pelo fenótipo de resistência na planta TE5-10-15J.Conselho Nacional de Desenvolvimento Científico e Tecnológicoapplication/pdfporUniversidade Federal de ViçosaDoutorado em FitopatologiaUFVBREtiologia; Epidemiologia; ControleMaracujáPotyvirusPlantas transgênicasPassionfruitPotyvirusTransgenic plantsCNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::FITOPATOLOGIAEstudo de isolados brasileiros de potyvírus causadores de endurecimento dos frutos do maracujazeiro e análise da resistência à doença em linhagens transgênicas R1 de maracujá-amareloStudy of Brazilian potyvirus isolates causing passionfruit woodiness and analysis of resistance to the disease in R1 transgenic lines of passionflowerinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/doctoralThesisinfo:eu-repo/semantics/openAccessreponame:LOCUS Repositório Institucional da UFVinstname:Universidade Federal de Viçosa (UFV)instacron:UFVORIGINAL01 - capa_literatura.pdfapplication/pdf265739https://locus.ufv.br//bitstream/123456789/1040/1/01%20-%20capa_literatura.pdf0417a96a6f8237c38b55b86dac8ab3dfMD5102 - capitulo 1.pdf02 - capitulo 1.pdfapplication/pdf229266https://locus.ufv.br//bitstream/123456789/1040/2/02%20-%20capitulo%201.pdfeb5b050fc055f3bbfccde6386c76173eMD5203 - conclusoes gerais.pdf03 - conclusoes gerais.pdfapplication/pdf74142https://locus.ufv.br//bitstream/123456789/1040/3/03%20-%20conclusoes%20gerais.pdfc745521c0007ecf702a60692f38cb8edMD53TEXT01 - capa_literatura.pdf.txt01 - capa_literatura.pdf.txtExtracted texttext/plain75390https://locus.ufv.br//bitstream/123456789/1040/4/01%20-%20capa_literatura.pdf.txta9d6958fbfeb22cf1145be53789c240dMD5402 - capitulo 1.pdf.txt02 - capitulo 1.pdf.txtExtracted texttext/plain32614https://locus.ufv.br//bitstream/123456789/1040/5/02%20-%20capitulo%201.pdf.txt02c5430add06dae2709e61b1b4f793abMD5503 - conclusoes gerais.pdf.txt03 - conclusoes gerais.pdf.txtExtracted texttext/plain1106https://locus.ufv.br//bitstream/123456789/1040/6/03%20-%20conclusoes%20gerais.pdf.txt2ae1d8c86c7f494503c36e040531e961MD56THUMBNAIL01 - capa_literatura.pdf.jpg01 - capa_literatura.pdf.jpgIM Thumbnailimage/jpeg3621https://locus.ufv.br//bitstream/123456789/1040/7/01%20-%20capa_literatura.pdf.jpg085d3afb0e1b6da044888a3e5d93ec3aMD5702 - capitulo 1.pdf.jpg02 - capitulo 1.pdf.jpgIM Thumbnailimage/jpeg3542https://locus.ufv.br//bitstream/123456789/1040/8/02%20-%20capitulo%201.pdf.jpg001600351d067b239a689f6f5af1ca65MD5803 - conclusoes gerais.pdf.jpg03 - conclusoes gerais.pdf.jpgIM Thumbnailimage/jpeg3949https://locus.ufv.br//bitstream/123456789/1040/9/03%20-%20conclusoes%20gerais.pdf.jpg8c7fb7fa4d68ee8d696d28fcbd22a5e3MD59123456789/10402016-04-06 23:17:25.94oai:locus.ufv.br:123456789/1040Repositório InstitucionalPUBhttps://www.locus.ufv.br/oai/requestfabiojreis@ufv.bropendoar:21452016-04-07T02:17:25LOCUS Repositório Institucional da UFV - Universidade Federal de Viçosa (UFV)false |
dc.title.por.fl_str_mv |
Estudo de isolados brasileiros de potyvírus causadores de endurecimento dos frutos do maracujazeiro e análise da resistência à doença em linhagens transgênicas R1 de maracujá-amarelo |
dc.title.alternative.eng.fl_str_mv |
Study of Brazilian potyvirus isolates causing passionfruit woodiness and analysis of resistance to the disease in R1 transgenic lines of passionflower |
title |
Estudo de isolados brasileiros de potyvírus causadores de endurecimento dos frutos do maracujazeiro e análise da resistência à doença em linhagens transgênicas R1 de maracujá-amarelo |
spellingShingle |
Estudo de isolados brasileiros de potyvírus causadores de endurecimento dos frutos do maracujazeiro e análise da resistência à doença em linhagens transgênicas R1 de maracujá-amarelo Nascimento, Ana Verônica Silva do Maracujá Potyvirus Plantas transgênicas Passionfruit Potyvirus Transgenic plants CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::FITOPATOLOGIA |
title_short |
Estudo de isolados brasileiros de potyvírus causadores de endurecimento dos frutos do maracujazeiro e análise da resistência à doença em linhagens transgênicas R1 de maracujá-amarelo |
title_full |
Estudo de isolados brasileiros de potyvírus causadores de endurecimento dos frutos do maracujazeiro e análise da resistência à doença em linhagens transgênicas R1 de maracujá-amarelo |
title_fullStr |
Estudo de isolados brasileiros de potyvírus causadores de endurecimento dos frutos do maracujazeiro e análise da resistência à doença em linhagens transgênicas R1 de maracujá-amarelo |
title_full_unstemmed |
Estudo de isolados brasileiros de potyvírus causadores de endurecimento dos frutos do maracujazeiro e análise da resistência à doença em linhagens transgênicas R1 de maracujá-amarelo |
title_sort |
Estudo de isolados brasileiros de potyvírus causadores de endurecimento dos frutos do maracujazeiro e análise da resistência à doença em linhagens transgênicas R1 de maracujá-amarelo |
author |
Nascimento, Ana Verônica Silva do |
author_facet |
Nascimento, Ana Verônica Silva do |
author_role |
author |
dc.contributor.authorLattes.por.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4760240Y3 |
dc.contributor.author.fl_str_mv |
Nascimento, Ana Verônica Silva do |
dc.contributor.advisor-co1.fl_str_mv |
Otoni, Wagner Campos |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4786133Y6 |
dc.contributor.advisor1.fl_str_mv |
Zerbini Júnior, Francisco Murilo |
dc.contributor.advisor1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4783743U5 |
dc.contributor.referee1.fl_str_mv |
Pio-ribeiro, Gilvan |
dc.contributor.referee1Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4793337P7 |
dc.contributor.referee2.fl_str_mv |
Zerbini, Poliane Alfenas |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/8632328159533071 |
dc.contributor.referee3.fl_str_mv |
Oliveira, Rosângela D arc de Lima |
dc.contributor.referee3Lattes.fl_str_mv |
http://buscatextual.cnpq.br/buscatextual/visualizacv.do?id=K4788026Z3 |
contributor_str_mv |
Otoni, Wagner Campos Zerbini Júnior, Francisco Murilo Pio-ribeiro, Gilvan Zerbini, Poliane Alfenas Oliveira, Rosângela D arc de Lima |
dc.subject.por.fl_str_mv |
Maracujá Potyvirus Plantas transgênicas |
topic |
Maracujá Potyvirus Plantas transgênicas Passionfruit Potyvirus Transgenic plants CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::FITOPATOLOGIA |
dc.subject.eng.fl_str_mv |
Passionfruit Potyvirus Transgenic plants |
dc.subject.cnpq.fl_str_mv |
CNPQ::CIENCIAS AGRARIAS::AGRONOMIA::FITOSSANIDADE::FITOPATOLOGIA |
description |
Passionfruit woodiness is the most important disease of passion flower. The disease can be caused by the potyviruses Passionfruit woodiness virus (PWV), Cowpea aphid-borne mosaic virus (CABMV) and East Asian Passiflora virus (EAPV). Infected plants display mosaic and woodiness of the fruit. In Brazil, the disease occurs in all states where passion flower is grown, and the causeal agent has always been identified as PWV based on biological and serological properties. Management strategies based on vector control with insecticides, cross-protection and natural resistance are inneficient. The objectives of this work were to characterize at the molecular level a number of Brazilian isolates of potyvirus causing passionfruit woodiness and to evaluate the resistance of transgenic yellow passion flower (Passiflora edulis f. flavicarpa) plants expressing a genomic fragment from a woodinessinducing potyvirus isolate from Brazil. Foliar samples of yellow passion flower showing typical symptoms of the disease were collected collected in the states of Bahia, Espírito Santo, Minas Gerais, Paraíba, Pernambuco and Sergipe. The presence of the virus was confirmed by serology and host range. The viral isolates were capable of infecting plants of common bean and cowpea, although differing in the intensity of symptoms induced in these hosts. Serological relationship between the potyvirus isolates from passion flower Cowpea aphidborne mosaic virus (CABMV) was demonstrated by indirect ELISA. The sequence of the capsid protein coding region was determined for all isolates. Sequence comparisons with other potyviruses indicated a level of identify from 85 to 94% with CABMV isolates. Identity with PWV isolate K ranged from 54 to 70%. Phylogenetic analysis based on the deduced capsid protein amino acid sequences clustered the Brazilian isolates together with CABMV isolates, distant from PWV isolates. Together, these results indicated that the potyvirus isolates from passion flower represent a strain of CABMV. In an attempt to effectively control passionfruit woodiness, transgenic yellow passionfruit plants (Passiflora edulis f. flavicarpa) were generated which express an untranslatable RNA corresponding to the replicase and capsid protein coding regions of a CABMV isolate from Minas Gerais (MGAvr). One R0 plant (TE5-10) was resistant to the MG-Avr isolate due to RNA silencing. However, this plant was susceptible to a second isolate of the virus (PE-Bnt). In order to verify the effect of gene dosage in the spectrum of resistance, several crosses were carried out between the TE5-10 plant, a susceptible R0 plant (T2-5) ans a non-transformed plant. Also, self-pollination of the TE5-10 plant was carried out in orde to generate transgene-homozigous R1 plants. The plants obtained from these crosses were challenged with three CABMV isolates (MG-Avr, PE-Bnt and SE-Nps). One R1 plant obtained from self-pollination (named TE5-10-15J) was resistant to the three isolates. This plant was challenged with an additional four CABMV isolates (PB-Alh, BA-Itb, ES-Vni and PA-Iga), being resistant to all of them based on symptom development. Northern analysis demonstrated the lack of transgenic mRNA accumulation in these plants, indicating that resistance is due to RNA silencing. |
publishDate |
2006 |
dc.date.issued.fl_str_mv |
2006-08-31 |
dc.date.available.fl_str_mv |
2007-02-15 2015-03-26T12:41:57Z |
dc.date.accessioned.fl_str_mv |
2015-03-26T12:41:57Z |
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info:eu-repo/semantics/publishedVersion |
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info:eu-repo/semantics/doctoralThesis |
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doctoralThesis |
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publishedVersion |
dc.identifier.citation.fl_str_mv |
NASCIMENTO, Ana Verônica Silva do. Study of Brazilian potyvirus isolates causing passionfruit woodiness and analysis of resistance to the disease in R1 transgenic lines of passionflower. 2006. 55 f. Tese (Doutorado em Etiologia; Epidemiologia; Controle) - Universidade Federal de Viçosa, Viçosa, 2006. |
dc.identifier.uri.fl_str_mv |
http://locus.ufv.br/handle/123456789/1040 |
identifier_str_mv |
NASCIMENTO, Ana Verônica Silva do. Study of Brazilian potyvirus isolates causing passionfruit woodiness and analysis of resistance to the disease in R1 transgenic lines of passionflower. 2006. 55 f. Tese (Doutorado em Etiologia; Epidemiologia; Controle) - Universidade Federal de Viçosa, Viçosa, 2006. |
url |
http://locus.ufv.br/handle/123456789/1040 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal de Viçosa |
dc.publisher.program.fl_str_mv |
Doutorado em Fitopatologia |
dc.publisher.initials.fl_str_mv |
UFV |
dc.publisher.country.fl_str_mv |
BR |
dc.publisher.department.fl_str_mv |
Etiologia; Epidemiologia; Controle |
publisher.none.fl_str_mv |
Universidade Federal de Viçosa |
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