Application of FTA technology for sampling, recovery and molecular characterization of viral pathogens and virusderived transgenes from plant tissues

Background: Plant viral diseases present major constraints to crop production. Effective sampling of the viruses infecting plants is required to facilitate their molecular study and is essential for the development of crop protection and improvement programs. Retaining integrity of viral pathogens w...

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Autores principales: Ndunguru, J., Taylor, N., Yadav, J.S., Aly, H., Legg, James P., Aveling, T., Thompson, G., Fauquet, Claude M.
Formato: Journal Article
Lenguaje:Inglés
Publicado: Springer 2005
Materias:
Acceso en línea:https://hdl.handle.net/10568/91505
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author Ndunguru, J.
Taylor, N.
Yadav, J.S.
Aly, H.
Legg, James P.
Aveling, T.
Thompson, G.
Fauquet, Claude M.
author_browse Aly, H.
Aveling, T.
Fauquet, Claude M.
Legg, James P.
Ndunguru, J.
Taylor, N.
Thompson, G.
Yadav, J.S.
author_facet Ndunguru, J.
Taylor, N.
Yadav, J.S.
Aly, H.
Legg, James P.
Aveling, T.
Thompson, G.
Fauquet, Claude M.
author_sort Ndunguru, J.
collection Repository of Agricultural Research Outputs (CGSpace)
description Background: Plant viral diseases present major constraints to crop production. Effective sampling of the viruses infecting plants is required to facilitate their molecular study and is essential for the development of crop protection and improvement programs. Retaining integrity of viral pathogens within sampled plant tissues is often a limiting factor in this process, most especially when sample sizes are large and when operating in developing counties and regions remote from laboratory facilities. FTA is a paper-based system designed to fix and store nucleic acids directly from fresh tissues pressed into the treated paper. We report here the use of FTA as an effective technology for sampling and retrieval of DNA and RNA viruses from plant tissues and their subsequent molecular analysis. Results: DNA and RNA viruses were successfully recovered from leaf tissues of maize, cassava, tomato and tobacco pressed into FTA® Classic Cards. Viral nucleic acids eluted from FTA cards were found to be suitable for diagnostic molecular analysis by PCR-based techniques andrestriction analysis, and for cloning and nucleotide sequencing in a manner equivalent to that offered by tradition isolation methods. Efficacy of the technology was demonstrated both from sampled greenhouse-grown plants and from leaf presses taken from crop plants growing in farmer's fields in East Africa. In addition, FTA technology was shown to be suitable for recovery of viral- derived transgene sequences integrated into the plant genome. Conclusion: Results demonstrate that FTA is a practical, economical and sensitive method for sampling, storage and retrieval of viral pathogens and plant genomic sequences, when working under controlled conditions and in the field. Application of this technology has the potential to significantly increase ability to bring modern analytical techniques to bear on the viral pathogens infecting crop plants.
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spelling CGSpace915052024-06-26T09:37:04Z Application of FTA technology for sampling, recovery and molecular characterization of viral pathogens and virusderived transgenes from plant tissues Ndunguru, J. Taylor, N. Yadav, J.S. Aly, H. Legg, James P. Aveling, T. Thompson, G. Fauquet, Claude M. molecular dna and rna viruses viral pathogens crop plants genomes germplasm Background: Plant viral diseases present major constraints to crop production. Effective sampling of the viruses infecting plants is required to facilitate their molecular study and is essential for the development of crop protection and improvement programs. Retaining integrity of viral pathogens within sampled plant tissues is often a limiting factor in this process, most especially when sample sizes are large and when operating in developing counties and regions remote from laboratory facilities. FTA is a paper-based system designed to fix and store nucleic acids directly from fresh tissues pressed into the treated paper. We report here the use of FTA as an effective technology for sampling and retrieval of DNA and RNA viruses from plant tissues and their subsequent molecular analysis. Results: DNA and RNA viruses were successfully recovered from leaf tissues of maize, cassava, tomato and tobacco pressed into FTA® Classic Cards. Viral nucleic acids eluted from FTA cards were found to be suitable for diagnostic molecular analysis by PCR-based techniques andrestriction analysis, and for cloning and nucleotide sequencing in a manner equivalent to that offered by tradition isolation methods. Efficacy of the technology was demonstrated both from sampled greenhouse-grown plants and from leaf presses taken from crop plants growing in farmer's fields in East Africa. In addition, FTA technology was shown to be suitable for recovery of viral- derived transgene sequences integrated into the plant genome. Conclusion: Results demonstrate that FTA is a practical, economical and sensitive method for sampling, storage and retrieval of viral pathogens and plant genomic sequences, when working under controlled conditions and in the field. Application of this technology has the potential to significantly increase ability to bring modern analytical techniques to bear on the viral pathogens infecting crop plants. 2005-12 2018-03-07T11:26:06Z 2018-03-07T11:26:06Z Journal Article https://hdl.handle.net/10568/91505 en Open Access Springer Ndunguru, J., Taylor, N. J., Yadav, J., Aly, H., Legg, J. P., Aveling, T., Thompson, G., & Fauquet, C. M. (2005). Application of FTA technology for sampling, recovery and molecular characterization of viral pathogens and virus-derived transgenes from plant tissues. Virology Journal, 2(1). https://doi.org/10.1186/1743-422x-2-45
spellingShingle molecular
dna and rna viruses
viral pathogens
crop plants
genomes
germplasm
Ndunguru, J.
Taylor, N.
Yadav, J.S.
Aly, H.
Legg, James P.
Aveling, T.
Thompson, G.
Fauquet, Claude M.
Application of FTA technology for sampling, recovery and molecular characterization of viral pathogens and virusderived transgenes from plant tissues
title Application of FTA technology for sampling, recovery and molecular characterization of viral pathogens and virusderived transgenes from plant tissues
title_full Application of FTA technology for sampling, recovery and molecular characterization of viral pathogens and virusderived transgenes from plant tissues
title_fullStr Application of FTA technology for sampling, recovery and molecular characterization of viral pathogens and virusderived transgenes from plant tissues
title_full_unstemmed Application of FTA technology for sampling, recovery and molecular characterization of viral pathogens and virusderived transgenes from plant tissues
title_short Application of FTA technology for sampling, recovery and molecular characterization of viral pathogens and virusderived transgenes from plant tissues
title_sort application of fta technology for sampling recovery and molecular characterization of viral pathogens and virusderived transgenes from plant tissues
topic molecular
dna and rna viruses
viral pathogens
crop plants
genomes
germplasm
url https://hdl.handle.net/10568/91505
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