Submitted to: American Phytopathological Society Annual Meeting
Publication Type: Abstract Only
Publication Acceptance Date: March 15, 2003
Publication Date: August 9, 2003
Citation: Hilf, M. E. 2003. Overlapping Primers for Cloning of the Citrus Tristeza Virus Genome. American Phytopathological Society Annual Meeting. Technical Abstract: Citrus tristeza virus (CTV) is the most prevalent and destructive viral pathogen of citrus. Genomic sequences of CTV isolates from different locations and with different biological properties have a highly conserved nucleotide sequence in the 3-prime eight kilobases of the genome, but a higher level of nucleotide sequence diversity in the 5-prime eleven kilobases. Conserved nucleotide sequences were identified from the alignment of the complete genomes of the pathogenically and genetically distinct T36, T30, VT and SY568 CTV isolates, and used to design primers for the polymerase chain reaction mediated cloning of sequences from CTV isolates of unknown genetic complexity. Overlapping primers designed to amplify a complete CTV genome were successfully used to amplify portions of the genomes of the Florida isolates T3 and T68, and from isolates from Peru, New Zealand and Japan. Sequence analysis of clones of T3 and T68 indicated these isolates were genetically distinct from those known, and can be considered new strains. The results suggest that this experimental approach can provide DNA for sequence analysis from genetically diverse CTV isolates.