Submitted to: Methods in Molecular Biology
Publication Type: Book / Chapter
Publication Acceptance Date: November 2, 2007
Publication Date: December 3, 2007
Citation: Huang, Q., Hartung, J.S. 2007. Construction of infectious clones for double stranded dna viruses of plants using citrus yellow mosaic cirus as an example. Methods in Molecular Biology. 451:525-534. Interpretive Summary: In order to control diseases caused by plant viruses, researchers must understand the mechanisms used by the virus to infect, replicate and induce symptoms in plants. By using full length infectious clones of the virus, mutants can be created and used to follow these events in plants. This article describes the procedures needed to produce a full length, infectious clone of a double stranded DNA virus. Researchers in the plant virology field will find this information helpful to advance their own research projects.
Technical Abstract: Double-stranded DNA (dsDNA) viruses of plants are believed to be plant pararetroviruses. Their genome is replicated by reverse transcription of a larger than unit length terminally redundant RNA transcript of the viral genomic DNA using the virus-encoded replicase. In order to produce a cloned, infectious viral genome, the clone must be constructed in a binary vector and be longer than the full, unit-length viral genome. The clone can then be transferred by Agrobacterium-assisted inoculation into a suitable host plant to induce virus infection.