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THE PLANT CELL, Vol 3, Issue 2 149-157, Copyright © 1991 by American Society of Plant Biologists
Embryonic Lethals and T-DNA Insertional Mutagenesis in Arabidopsis
D. Errampalli, D. Patton, L. Castle, L. Mickelson, K. Hansen, J. Schnall, K. Feldmann and D. Meinke
Department of Botany, Oklahoma State University, Stillwater, Oklahoma 74078
T-DNA insertional mutagenesis represents a promising approach to the
molecular isolation of genes with essential functions during plant embryo
development. We describe in this report the isolation and characterization
of 18 mutants of Arabidopsis thaliana defective in embryo development
following seed transformation with Agrobacterium tumefaciens. Random T-DNA
insertion was expected to result in a high frequency of recessive embryonic
lethals because many target genes are required for embryogenesis. The
cointegrate Ti plasmid used in these experiments contained the nopaline
synthase and neomycin phosphotransferase gene markers. Nopaline assays and
resistance to kanamycin were used to estimate the number of functional
inserts present in segregating families. Nine families appeared to contain
a T-DNA insert either within or adjacent to the mutant gene. Eight families
were clearly not tagged with a functional insert and appeared instead to
contain mutations induced during the transformation process. DNA gel blot
hybridization with internal and right border probes revealed a variety of
rearrangements associated with T-DNA insertion. A general strategy is
presented to simplify the identification of tagged embryonic mutants and
facilitate the molecular isolation of genes required for plant
embryogenesis.
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