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THE PLANT CELL, Vol 1, Issue 11 1057-1067, Copyright © 1989 by American Society of Plant Biologists
Functional Expression of the Leftward Open Reading Frames of the A Component of Tomato Golden Mosaic Virus in Transgenic Tobacco Plants
L. Hanley-Bowdoin, J. S. Elmer and S. G. Rogers
Plant Gene Expression, Corporate Research Laboratories, Monsanto Company, 700 Chesterfield Village Parkway, St. Louis, Missouri 63198
The genome of the geminivirus tomato golden mosaic virus (TGMV) consists of
two circular DNA molecules designated as components A and B. We have
constructed Nicotiana benthamiana plants that are transgenic for the three
overlapping open reading frames, AL1, AL2, and AL3, from the left side of
TGMV A. In the transgenic plants, the AL open reading frames are under the
control of the cauliflower mosaic virus (CaMV) 35S promoter. In TGMV
infectivity assays, seven of 10 transgenic lines complemented TGMV A
variants with mutations in AL1, AL2, or AL3 when co-inoculated with the B
component. The 35S-AL construct was transcribed as a single RNA species in
the transgenic plants, indicating that AL1, AL2, and AL3 were expressed
from a polycistronic mRNA. This differs from the complex transcription
pattern in TGMV-infected plants, which contains five AL transcripts. There
was no quantitative correlation between the efficiency of complementation
in the infectivity assay and the level of expression of transgenic AL RNA
in the leaves of a transgenic line. One line that failed to complement
defects in AL1, AL2, and AL3 in infectivity assays contained high levels of
transgenic AL RNA and functional AL1 protein. These results provide
evidence that chromosomal position can affect the cell- and tissue-specific
transcription of the 35S promoter in transgenic plants. Comparison of the
complementing plants and wild-type infected plants may provide insight into
the TGMV infection process and the use of the CaMV 35S promoter for gene
expression in transgenic plants.
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