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Agroinfection of Nicotiana spp. with Cloned DNA of Tomato Golden Mosaic Virus

Department of Pure and Applied Biology, Imperial College of Science and Technology, Prince Consort Road, London SW7 2BB, U.K.

Head-to-tail dimers of cloned DNA A and DNA B of tomato golden mosaic virus (TGMV) were integrated between the T-DNA border sequences of a broad host range binary vector and transferred into cells of Nicotiana benthamiana seedlings using an Agrobacterium tumefaciens-mediated delivery system. Most of the inoculated plants developed golden-yellow mosaic and leaf curling symptoms typical of TGMV infection. Extracts of infected leaves were shown to contain both double-stranded and single-stranded TGMV DNA forms of genome length and the virus capsid polypeptide. Infection was also achieved by inoculating plants with mixtures of Agrobacterium strains containing dimers of DNA A or DNA B, with a strain containing a partial dimer of DNA A and a dimer of DNA B and with a strain containing a dimer of DNA B and a partial dimer of DNA A with a 603 bp deletion in the coat protein gene. In the latter case, mosaic symptoms were mild and leaves did not curl. Transgenic N. tabacum cv. Samsun plants containing head-to-tail dimers of DNA A (A2 plants) or DNA B (B2 plants) were produced by transformation with Ti plasmid vectors. A2 plants and B2 plants were agroinfected with dimeric DNA B and dimeric DNA A, respectively. In both cases, symptoms typical of TGMV infection were induced and viral single-stranded DNA of both components was detected in the systemically infected tissue.

Keywords: TGMV, geminivirus, agroinfection

Received 26 January 1988; accepted 10 March 1988.