Characterization of cucumber mosaic virus. VI. Generation of deletions in defective RNA 3s during passage in transgenic tobacco expressing the 3a gene.

Defective mutants of cucumber mosaic virus (CMV) RNA 3, containing deletions in the 3a gene were passaged in transgenic tobacco plants expressing the CMV 3a gene. After six passages, the various progeny RNA 3 were characterized. In all but one case, the size of the deletion increased. For the NheI-fs RNA 3 mutant of the Fny-strain of CMV (with an 8 nucleotide deletion), the deletion increased in the progeny viral RNA 3 to 570 nucleotides. For a similar frameshift mutant in RNA 3 of the M strain of CMV, either single RNA 3 species (with deletions of 579 or 627 nucleotides) or mixtures of RNA 3 deletion variants were observed in different plants. The DeltaE-H mutant (with a deletion of 202 nucleotides) of Fny-CMV RNA 3 underwent further deletion resulting in the loss of the entire 3a gene and flanking sequences. The DeltaKpnI mutant (deletion of 501 nucleotides) of Fny-CMV RNA 3 underwent a further deletion of 30 nucleotides. Except for the deletion progeny of the DeltaE-H RNA 3 mutant, the other defective RNA 3s all contained inframe deletions. It is proposed that the various deletions were created by different types of recombination and that packaging may be an important factor in the selection of particular defective RNA 3 variants.