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

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    Date
    2005
    Author
    Ndunguru, J.
    Taylor, N.
    Yadav, J.S.
    Aly, H.
    Legg, J.P.
    Aveling, T.
    Thompson, G.
    Fauquet, Claude M.
    Type
    Journal Article
    Metadata
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    Abstract/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.
    https://dx.doi.org/10.1186/1743-422X-2-45
    Multi standard citation
    Permanent link to this item
    https://hdl.handle.net/20.500.12478/3182
    Digital Object Identifier (DOI)
    https://dx.doi.org/10.1186/1743-422X-2-45
    IITA Subjects
    Plant Genetic Resources; Pests Of Plants; Plant Breeding; Diseases Control; Farm Management; Food Security; Genetic Improvement; Plant Diseases; Plant Production
    Agrovoc Terms
    Molecular; Dna And Rna Viruses; Viral Pathogens; Crop Plants; Genomes; Germplasm
    Regions
    Acp; Africa; North America; Middle East; East Africa; Southern Africa
    Countries
    United States; Tanzania; Egypt; Uganda; South Africa
    Collections
    • Journal and Journal Articles4836
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