Optimizing conditions for biolistic inoculation of Begomovirus in tomato and tobacco

Experimental transmission of Begomovirus is problematic. Most Begomoviruses can be transmitted readily from plant to plant by the whitefly vector, but this also requires a live infected plant and extensive facilities to maintain the insect. Whitefly inoculations can also be problematic because of their preferential feeding habits on certain plants. Mechanical inoculation of Begomovirus is possible but generally at low rates and for others not at all. For this reason particle bombardment (biolistic) of DNA viral as an inoculum was developed. The possibility of using the Helios Gen Gun System(Biorad®), a biolistic hand-held device, for transmitting Begomovirus bipartite to tomato and tobacco plants was assayed and optimized. Biolistic inoculation was carried out with the hand held device at 220 or 320 psi, applying 1 or 2 shots /plant and using gold particles of 0.6 or 1.6µm in size. Characteristic symptoms of viral disease (chlorosis, mosaic and leaf deformation) appeared 3 weeks post-inoculation in the newly developing leaves. Replication of the viral DNA in plants was confirmed by Polymerase Chain Reaction. All bombarded plants became infected when biolistic inoculation was carried out with the hand held device at 320psi and using 1.6 µm gold particles in size. To our knowledge this is the first report in Colombia of successful direct inoculation of tomato and tobacco plants with Begomovirus bipartite geminivirus using a biolistic hand-held device.

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Main Authors: López-López, Karina, Rodríguez-Mora, Diana Milena, Vaca-Vaca, Juan Carlos
Format: Digital revista
Language:spa
Published: Universidad Nacional de Colombia - Sede Bogotá - Instituto de Biotecnología 2013
Online Access:https://revistas.unal.edu.co/index.php/biotecnologia/article/view/41261
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institution UNAL
collection OJS
country Colombia
countrycode CO
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databasecode rev-biotecnologia-co
tag revista
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language spa
format Digital
author López-López, Karina
Rodríguez-Mora, Diana Milena
Vaca-Vaca, Juan Carlos
spellingShingle López-López, Karina
Rodríguez-Mora, Diana Milena
Vaca-Vaca, Juan Carlos
Optimizing conditions for biolistic inoculation of Begomovirus in tomato and tobacco
author_facet López-López, Karina
Rodríguez-Mora, Diana Milena
Vaca-Vaca, Juan Carlos
author_sort López-López, Karina
title Optimizing conditions for biolistic inoculation of Begomovirus in tomato and tobacco
title_short Optimizing conditions for biolistic inoculation of Begomovirus in tomato and tobacco
title_full Optimizing conditions for biolistic inoculation of Begomovirus in tomato and tobacco
title_fullStr Optimizing conditions for biolistic inoculation of Begomovirus in tomato and tobacco
title_full_unstemmed Optimizing conditions for biolistic inoculation of Begomovirus in tomato and tobacco
title_sort optimizing conditions for biolistic inoculation of begomovirus in tomato and tobacco
description Experimental transmission of Begomovirus is problematic. Most Begomoviruses can be transmitted readily from plant to plant by the whitefly vector, but this also requires a live infected plant and extensive facilities to maintain the insect. Whitefly inoculations can also be problematic because of their preferential feeding habits on certain plants. Mechanical inoculation of Begomovirus is possible but generally at low rates and for others not at all. For this reason particle bombardment (biolistic) of DNA viral as an inoculum was developed. The possibility of using the Helios Gen Gun System(Biorad®), a biolistic hand-held device, for transmitting Begomovirus bipartite to tomato and tobacco plants was assayed and optimized. Biolistic inoculation was carried out with the hand held device at 220 or 320 psi, applying 1 or 2 shots /plant and using gold particles of 0.6 or 1.6µm in size. Characteristic symptoms of viral disease (chlorosis, mosaic and leaf deformation) appeared 3 weeks post-inoculation in the newly developing leaves. Replication of the viral DNA in plants was confirmed by Polymerase Chain Reaction. All bombarded plants became infected when biolistic inoculation was carried out with the hand held device at 320psi and using 1.6 µm gold particles in size. To our knowledge this is the first report in Colombia of successful direct inoculation of tomato and tobacco plants with Begomovirus bipartite geminivirus using a biolistic hand-held device.
publisher Universidad Nacional de Colombia - Sede Bogotá - Instituto de Biotecnología
publishDate 2013
url https://revistas.unal.edu.co/index.php/biotecnologia/article/view/41261
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spelling oai:www.revistas.unal.edu.co:article-412612019-07-02T21:22:54Z Optimizing conditions for biolistic inoculation of Begomovirus in tomato and tobacco Optimización de las condiciones de inoculación por biobalística de un Begomovirus en tomate y tabaco López-López, Karina Rodríguez-Mora, Diana Milena Vaca-Vaca, Juan Carlos geminivirus Solanum lycopersicum Nicotiana tabacum N benthamiana Bemisia tabaci geminivirus Solanum lycopersicum Nicotiana tabacum N. benthamiana Bemisia tabaci Experimental transmission of Begomovirus is problematic. Most Begomoviruses can be transmitted readily from plant to plant by the whitefly vector, but this also requires a live infected plant and extensive facilities to maintain the insect. Whitefly inoculations can also be problematic because of their preferential feeding habits on certain plants. Mechanical inoculation of Begomovirus is possible but generally at low rates and for others not at all. For this reason particle bombardment (biolistic) of DNA viral as an inoculum was developed. The possibility of using the Helios Gen Gun System(Biorad®), a biolistic hand-held device, for transmitting Begomovirus bipartite to tomato and tobacco plants was assayed and optimized. Biolistic inoculation was carried out with the hand held device at 220 or 320 psi, applying 1 or 2 shots /plant and using gold particles of 0.6 or 1.6µm in size. Characteristic symptoms of viral disease (chlorosis, mosaic and leaf deformation) appeared 3 weeks post-inoculation in the newly developing leaves. Replication of the viral DNA in plants was confirmed by Polymerase Chain Reaction. All bombarded plants became infected when biolistic inoculation was carried out with the hand held device at 320psi and using 1.6 µm gold particles in size. To our knowledge this is the first report in Colombia of successful direct inoculation of tomato and tobacco plants with Begomovirus bipartite geminivirus using a biolistic hand-held device. La transmisión experimental de Begomovirus es problemática. La mayoría de estos virus se pueden transmitir de planta a planta por su vector biológico, Bemisia tabaci. Las inoculaciones experimentales con mosca blanca son problemáticas debido a sus hábitos de alimentación, requerimiento de una planta viva infectada e instalaciones de contención para el vector. Por su parte la inoculación mecánica de Begomovirus es posible, pero generalmente a tasas bajas y no en todos los casos. Por esta razón el bombardeo de partículas (biobalística) de DNA viral como una estrategia de inoculación fue desarrollada. La posibilidad de utilizar el dispositivo de mano Helios Gen Gun System (Biorad®), un equipo de biobalística, para la transmisión de un Begomovirus bipartita a plantas de tomate y tabaco fue ensayado y optimizado. Los parámetros evaluados fueron: número de disparos (1-2), presión de helio (220 y 320 psi) y diámetro de las partículas de oro (0.6 y 1.6µm). Los síntomas característicos de la enfermedad viral (clorosis, mosaico y deformación de la hoja) aparecieron 3 semanas después del bombardeo en las hojas jóvenes no inoculadas. La replicación del DNA viral en las plantas se confirmó por Reacción en cadena de la polimerasa. Plantas infectadas en un 100% se obtuvieron cuando en el bombardeo se emplearon partículas de oro de 1.6 µm recubiertas con DNA viral a una presión de 320psi. A nuestro entender este es el primer reporte en Colombia de la inoculación directa de plantas de tomate y tabaco con un Begomovirus bipartita usando un dispositivo portátil de biobalística. Universidad Nacional de Colombia - Sede Bogotá - Instituto de Biotecnología 2013-07-01 info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion Artículo revisado por pares application/vnd.openxmlformats-officedocument.wordprocessingml.document application/pdf text/html https://revistas.unal.edu.co/index.php/biotecnologia/article/view/41261 10.15446/rev.colomb.biote.v15n2.41261 Revista Colombiana de Biotecnología; Vol. 15 No. 2 (2013); 8-17 Revista Colombiana de Biotecnología; Vol. 15 Núm. 2 (2013); 8-17 1909-8758 0123-3475 spa https://revistas.unal.edu.co/index.php/biotecnologia/article/view/41261/42826 https://revistas.unal.edu.co/index.php/biotecnologia/article/view/41261/43479 https://revistas.unal.edu.co/index.php/biotecnologia/article/view/41261/43793 Derechos de autor 2013 Revista Colombiana de Biotecnología https://creativecommons.org/licenses/by/4.0