Development of Viral Vectors Based on Citrus leaf blotch virus to Express Foreign Proteins or Analyze Gene Function in Citrus Plants

Viral vectors have been used to express foreign proteins in plants or to silence endogenous genes. This methodology could be appropriate for citrus plants that have long juvenile periods and adult plants that are difficult to transform. We developed viral vectors based on Citrus leaf blotch virus (C...

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Autores principales: Agüero, Jesús, Ruiz-Ruiz, Susana, Vives, María C., Velázquez, Karelia, Navarro, Luis, Pena, Leandro, Moreno, Pedro, Guerri, José
Formato: article
Lenguaje:Inglés
Publicado: 2017
Acceso en línea:http://hdl.handle.net/20.500.11939/4877
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author Agüero, Jesús
Ruiz-Ruiz, Susana
Vives, María C.
Velázquez, Karelia
Navarro, Luis
Pena, Leandro
Moreno, Pedro
Guerri, José
author_browse Agüero, Jesús
Guerri, José
Moreno, Pedro
Navarro, Luis
Pena, Leandro
Ruiz-Ruiz, Susana
Velázquez, Karelia
Vives, María C.
author_facet Agüero, Jesús
Ruiz-Ruiz, Susana
Vives, María C.
Velázquez, Karelia
Navarro, Luis
Pena, Leandro
Moreno, Pedro
Guerri, José
author_sort Agüero, Jesús
collection ReDivia
description Viral vectors have been used to express foreign proteins in plants or to silence endogenous genes. This methodology could be appropriate for citrus plants that have long juvenile periods and adult plants that are difficult to transform. We developed viral vectors based on Citrus leaf blotch virus (CLBV) by duplicating a minimum promoter (92 bp) either at the 3′ untranslated region (clbv3′pr vector) or at the intergenic region between the movement and coat protein (CP) genes (clbvINpr vector). The duplicated fragment (–42/+50) around the transcription start site of the CP subgenomic RNA (sgRNA) had the full promoter activity and induced synthesis of a new sgRNA in infected plants. Agroinoculation with these vectors resulted in systemic infection of Nicotiana benthamiana and the resulting virions systemically infected citrus plants. A clbvINpr vector carrying the green fluorescent protein (GFP) gene expressed GFP in citrus plants and triggered gfp silencing in gfp-transgenic citrus plants, and vectors carrying fragments of the phytoene desaturase or the magnesium chelatase genes incited a silencing phenotype in citrus plants. These silenced phenotypes persisted in successive flushes. Because CLBV infections are symptomless in most citrus species, the effective silencing induced by CLBV-derived vectors will be helpful to analyze citrus gene function.
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spelling ReDivia48772025-04-25T14:44:49Z Development of Viral Vectors Based on Citrus leaf blotch virus to Express Foreign Proteins or Analyze Gene Function in Citrus Plants Agüero, Jesús Ruiz-Ruiz, Susana Vives, María C. Velázquez, Karelia Navarro, Luis Pena, Leandro Moreno, Pedro Guerri, José Viral vectors have been used to express foreign proteins in plants or to silence endogenous genes. This methodology could be appropriate for citrus plants that have long juvenile periods and adult plants that are difficult to transform. We developed viral vectors based on Citrus leaf blotch virus (CLBV) by duplicating a minimum promoter (92 bp) either at the 3′ untranslated region (clbv3′pr vector) or at the intergenic region between the movement and coat protein (CP) genes (clbvINpr vector). The duplicated fragment (–42/+50) around the transcription start site of the CP subgenomic RNA (sgRNA) had the full promoter activity and induced synthesis of a new sgRNA in infected plants. Agroinoculation with these vectors resulted in systemic infection of Nicotiana benthamiana and the resulting virions systemically infected citrus plants. A clbvINpr vector carrying the green fluorescent protein (GFP) gene expressed GFP in citrus plants and triggered gfp silencing in gfp-transgenic citrus plants, and vectors carrying fragments of the phytoene desaturase or the magnesium chelatase genes incited a silencing phenotype in citrus plants. These silenced phenotypes persisted in successive flushes. Because CLBV infections are symptomless in most citrus species, the effective silencing induced by CLBV-derived vectors will be helpful to analyze citrus gene function. 2017-06-01T10:11:14Z 2017-06-01T10:11:14Z 2012 OCT 2012 article publishedVersion Agueero, Jesus, Ruiz-Ruiz, S., Vives, M.C., Velazquez, Karelia, Navarro, L., Pena, L., Moreno, P., Guerri, J. (2012). Development of Viral Vectors Based on Citrus leaf blotch virus to Express Foreign Proteins or Analyze Gene Function in Citrus Plants. Molecular Plant-Microbe Interactions, 25(10), 1326-1337. 0894-0282 http://hdl.handle.net/20.500.11939/4877 10.1094/MPMI-02-12-0048-R en openAccess Impreso
spellingShingle Agüero, Jesús
Ruiz-Ruiz, Susana
Vives, María C.
Velázquez, Karelia
Navarro, Luis
Pena, Leandro
Moreno, Pedro
Guerri, José
Development of Viral Vectors Based on Citrus leaf blotch virus to Express Foreign Proteins or Analyze Gene Function in Citrus Plants
title Development of Viral Vectors Based on Citrus leaf blotch virus to Express Foreign Proteins or Analyze Gene Function in Citrus Plants
title_full Development of Viral Vectors Based on Citrus leaf blotch virus to Express Foreign Proteins or Analyze Gene Function in Citrus Plants
title_fullStr Development of Viral Vectors Based on Citrus leaf blotch virus to Express Foreign Proteins or Analyze Gene Function in Citrus Plants
title_full_unstemmed Development of Viral Vectors Based on Citrus leaf blotch virus to Express Foreign Proteins or Analyze Gene Function in Citrus Plants
title_short Development of Viral Vectors Based on Citrus leaf blotch virus to Express Foreign Proteins or Analyze Gene Function in Citrus Plants
title_sort development of viral vectors based on citrus leaf blotch virus to express foreign proteins or analyze gene function in citrus plants
url http://hdl.handle.net/20.500.11939/4877
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