Development of a non-radioactive molecular hybridization probe for detecting Strawberry mottle virus in strawberry
The vegetative propagation of strawberries favors transmission of systemic pathogens, such as viruses, which are one of the main yield-limiting factors for this crop. More than 20 viruses have been described as infecting this species; one of the most frequent is the Strawberry mottle virus (SMoV), w...
| Autores principales: | , , , , |
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| Formato: | Artículo |
| Lenguaje: | Español |
| Publicado: |
2017
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| Materias: | |
| Acceso en línea: | http://hdl.handle.net/20.500.12123/1233 https://revistas.unc.edu.ar/index.php/agris/article/view/16570/16381 http://www.scielo.org.ar/pdf/agrisc/v33n1/v33n1a04.pdf |
| _version_ | 1855482852224794624 |
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| author | Asinari, Florencia Cafrune, Eva Encarnacion Guzman, Fabiana Conci, Luis Rogelio Conci, Vilma Cecilia |
| author_browse | Asinari, Florencia Cafrune, Eva Encarnacion Conci, Luis Rogelio Conci, Vilma Cecilia Guzman, Fabiana |
| author_facet | Asinari, Florencia Cafrune, Eva Encarnacion Guzman, Fabiana Conci, Luis Rogelio Conci, Vilma Cecilia |
| author_sort | Asinari, Florencia |
| collection | INTA Digital |
| description | The vegetative propagation of strawberries favors transmission of systemic pathogens, such as viruses, which are one of the main yield-limiting factors for this crop. More than 20 viruses have been described as infecting this species; one of the most frequent is the Strawberry mottle virus (SMoV), which is responsible for significant economic losses. SMoV is usually detected by reverse transcription polymerase chain reaction (RT-PCR), given that serum is not available for serological tests. In this study, a non-radioactive molecular probe was developed for SMoV detection. The cDNA was synthesized by RT-PCR using specific primers designed from the 3’UTR region of the viral genome. The cloned cDNA segment was labeled and used as a probe. Six RNA extraction protocols were evaluated, and the modified cetyltrimethylammonium bromide (CTAB) method showed the highest sensitivity level. Leaves at different phenological stages and petioles were evaluated; the highest reaction was observed in old leaves and in petioles |
| format | Artículo |
| id | INTA1233 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Español |
| publishDate | 2017 |
| publishDateRange | 2017 |
| publishDateSort | 2017 |
| record_format | dspace |
| spelling | INTA12332017-11-17T13:40:03Z Development of a non-radioactive molecular hybridization probe for detecting Strawberry mottle virus in strawberry Asinari, Florencia Cafrune, Eva Encarnacion Guzman, Fabiana Conci, Luis Rogelio Conci, Vilma Cecilia Enfermedades de las Plantas Fresa Hibridación Virus de las Plantas Plant Diseases Strawberries Hybridization Plant Viruses Frutilla Virus del Moteado de la Frutilla The vegetative propagation of strawberries favors transmission of systemic pathogens, such as viruses, which are one of the main yield-limiting factors for this crop. More than 20 viruses have been described as infecting this species; one of the most frequent is the Strawberry mottle virus (SMoV), which is responsible for significant economic losses. SMoV is usually detected by reverse transcription polymerase chain reaction (RT-PCR), given that serum is not available for serological tests. In this study, a non-radioactive molecular probe was developed for SMoV detection. The cDNA was synthesized by RT-PCR using specific primers designed from the 3’UTR region of the viral genome. The cloned cDNA segment was labeled and used as a probe. Six RNA extraction protocols were evaluated, and the modified cetyltrimethylammonium bromide (CTAB) method showed the highest sensitivity level. Leaves at different phenological stages and petioles were evaluated; the highest reaction was observed in old leaves and in petioles La propagación vegetativa del cultivo de frutilla favorece la transmisión de patógenos sistémicos, como es el caso de los virus, que constituyen uno de los principales factores limitantes. Se han descripto más de 20 virus que infectan esta especie; el Strawberry mottle virus (SMoV) es uno de los más frecuentes y responsable de importantes pérdidas económicas. Debido a la falta de antisuero disponible comercialmente para un diagnóstico serológico, el SMoV es detectado fundamentalmente mediante reacción en cadena de la polimerasa con transcripción reversa (RT-PCR). En este estudio se desarrolló una sonda de hibridación molecular no radioactiva para su detección. Se sintetizó cDNA con cebadores específicos diseñados a partir de la región 3’ no codificante del genoma viral. El cDNA obtenido fue clonado, marcado y utilizado como sonda. Se evaluaron seis protocolos de extracción de ARN viral a partir de plantas infectadas, de los cuales el método de bromuro de cetiltrimetilamonio modificado (CTAB) fue el más eficiente. Se evaluaron hojas de diferentes estados fenológicos y pecíolos, y fueron las hojas viejas y los pecíolos los que mostraron mayor reacción. Inst. Patología Vegetal Fil: Asinari, Florencia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina Fil:Cafrune, Eva Encarnacion. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina Fil: Guzman, Fabiana Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina Fil: Conci, Luis Rogelio. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina. Fil: Conci, Vilma Cecilia. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Patología Vegetal; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2017-09-18T12:21:21Z 2017-09-18T12:21:21Z 2016 info:eu-repo/semantics/article info:ar-repo/semantics/artículo info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/1233 https://revistas.unc.edu.ar/index.php/agris/article/view/16570/16381 http://www.scielo.org.ar/pdf/agrisc/v33n1/v33n1a04.pdf 1668-298X (Online) spa info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Agriscientia [online] 33 (1) : 39-45 (2016) |
| spellingShingle | Enfermedades de las Plantas Fresa Hibridación Virus de las Plantas Plant Diseases Strawberries Hybridization Plant Viruses Frutilla Virus del Moteado de la Frutilla Asinari, Florencia Cafrune, Eva Encarnacion Guzman, Fabiana Conci, Luis Rogelio Conci, Vilma Cecilia Development of a non-radioactive molecular hybridization probe for detecting Strawberry mottle virus in strawberry |
| title | Development of a non-radioactive molecular hybridization probe for detecting Strawberry mottle virus in strawberry |
| title_full | Development of a non-radioactive molecular hybridization probe for detecting Strawberry mottle virus in strawberry |
| title_fullStr | Development of a non-radioactive molecular hybridization probe for detecting Strawberry mottle virus in strawberry |
| title_full_unstemmed | Development of a non-radioactive molecular hybridization probe for detecting Strawberry mottle virus in strawberry |
| title_short | Development of a non-radioactive molecular hybridization probe for detecting Strawberry mottle virus in strawberry |
| title_sort | development of a non radioactive molecular hybridization probe for detecting strawberry mottle virus in strawberry |
| topic | Enfermedades de las Plantas Fresa Hibridación Virus de las Plantas Plant Diseases Strawberries Hybridization Plant Viruses Frutilla Virus del Moteado de la Frutilla |
| url | http://hdl.handle.net/20.500.12123/1233 https://revistas.unc.edu.ar/index.php/agris/article/view/16570/16381 http://www.scielo.org.ar/pdf/agrisc/v33n1/v33n1a04.pdf |
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