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Infectious and Non-infectious Mutants of Cauliflower Mosaic Virus DNA

David L. Steffens

David J. Lyttle

Haifan Lin

Department of Biochemistry, Oklahoma State University, Stillwater, Oklahoma 74078, U.S.A.
and¹ Institute of Molecular and Cellular Biology, CNRS, 67084 Strasbourg Cedex, France

Mutants of cauliflower mosaic virus (CaMV), generated in vitro by modification of recombinant DNA plasmids containing the viral genome, either retained the ability to induce disease symptoms on turnip plants, produced less severe symptoms or failed to induce symptoms. Wild-type symptoms were produced by a variant CaMV DNA of the Cabbage S isolate that had 4 bp in open reading frame (ORF) III replaced with a 16 bp sequence. Less severe symptoms, due to a delay in symptom appearance relative to inoculation with wild-type DNA, were induced by a mutant with a frameshift mutation in ORF II (pSA103). CaMV DNA, recovered from plants infected with pSA103, contained a second mutation which restored the original translation reading frame. Nucleic acid hybridization to ‘squishes’ of leaf tissue from plants that had been inoculated with mutant DNAs that included DNAs modified in each of the six major ORFs of CaMV DNA revealed that only those plants that appeared diseased had detectable CaMV nucleic acid in uninoculated leaves. Replicated CaMV DNA was also not detected in non-encapsidated and virion DNA fractions from inoculated leaves of non-diseased plants.

Keywords: caulimoviruses, frameshift, DNA hybridization, mutants

Present address: Sigma Chemical Co., P.O. Box 14508, St. Louis, Missouri, U.S.A.

Present address: N.R.C. Plant Biotechnology Institute, 110 Gymnasium Rd., Saskatoon, Saskatchewan, Canada.

Present address: Program inGenetics, Cornell University, Ithaca, New York, U.S.A.

^< Present address: Division of Allergy and Immunology, University of Tennessee Medical School, Memphis, Tennessee, U.S.A.

Received 23 October 1985; accepted 24 March 1986.

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