THE PLANT CELL, Vol 3, Issue 3 247-258, Copyright © 1991 by American Society of Plant Biologists
Wheat Dwarf Virus Vectors Replicate and Express Foreign Genes in Cells of Monocotyledonous Plants
V. Matzeit, S. Schaefer, M. Kammann, H. J. Schalk, J. Schell and B. Gronenborn
Max-Planck-Institut fur Zuchtungsforschung, Abt. Genetische Grundlagen der Pflanzenzuchtung, Carl-von-Linne-Weg 10, D-5000 Koln 30, Federal Republic of Germany
Wheat dwarf virus (WDV) is a geminivirus that infects monocotyledonous
plants. To exploit the potential of WDV as a replicative gene vector, we
developed a transient replication and expression system based on the
transfection of protoplasts derived from Triticum monococcum suspension
culture cells. Cloned genomic copies of various WDV isolates as well as
mutants constructed in vitro were introduced into the protoplasts and
assayed for their ability to replicate. As a result, regions of the WDV
genome necessary or dispensable for the viral DNA replication could be
defined. In addition, the gene encoding the viral capsid protein was
replaced by three different bacterial marker genes, neomycin
phosphotransferase, chloramphenicol acetyltransferase, and
[beta]-galactosidase. The [beta]-galactosidase gene doubled the size of the
WDV genome. The replication of the recombinant WDV genomes and the
expression of these genes were monitored in suspension culture cells of T.
monococcum. The potential of replicative expression vectors based on the
WDV genome is discussed.
