Citrus tristeza virus (CTV) resistance in transgenic citrus based on virus challenge of protoplasts |
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Authors: | O?Olivares-Fuster G?H?Fleming M?R?Albiach-Marti S?Gowda W?O?Dawson Email author" target="_blank">J?W?CrosserEmail author |
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Institution: | (1) Horticulture Department, University of Florida-IFAS, Citrus Research and Education Center, 700 Experiment Station Road, 33850 Lake Alfred, FL |
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Abstract: | Summary A strategy for the sereening of candidate virus-derived sequences to provide RNA-mediated citrus tristeza virus (CTV) resistance
and early selection of virus-resistant citrus is presented. The system is based on the polyethylene glycol-(PEG) mediated
cotransformation of protoplasts using virus-derived sequences and green fluorescent protein as a single selectable marker,
followed by an in vitro assay of virus inoculation into transgenic protoplasts to determine the level of citrus tristeza virus replication. A cotransformation
rate higher than 20% allowed selection of several clones carrying the desired transgenes. Efficient in vitro inoculation of virus in transgenic protoplasts was performed. Tobacco mosaic virus virions were used as a control in order
to check eitrus protoplast viability. Different CTV replication levels were detected in transgenic clones. Only one clone
showed no replication of CTV. Considerations regarding selection of candidate virusderived sequences and virus challenge of
transgenic cells are presented. |
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Keywords: | eitrus protoplasts citrus tristeza virus PEG transformation virus inoculation virus resistance |
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