Two different types of T-DNA insert were found in tobacco plants transformed with
Agrobacterium tumefaciens. High-expressing (H) types had one copy of the T-DNA at a locus and produced high expression of the transgene
uidA, as measured by
uidA RNA levels and

-glucuronidase activity; low-expressing (L) types had inverted repeats of the T-DNA at a locus and produced low
uidA expression. H-types from different transformants acted additively, and cross-fertilization between two different homozygous transformants with H-type inserts produced F
1 plants with GUS activity that equalled the parents and individual F
2 plants with 50%, 100%, 150% and 200% of parental values. However, the L-type inserts worked
in trans to suppress
uidA expression from H-type inserts when both were present in the same genome. Hence when a transformant homozygous for the L-type insert was crossed to one homozygous for the H-type, all plants in the F
1 and F
2 generations with both types of insert had low GUS activity while F
2 segregants that only had the H-type inserts had high GUS activity again. Suppression of the H-type gene was associated with increased methylation of the insert. Particle acceleration was used to introduce further copies of
uidA into tissues of the transformants. Regardless of the promoter used, those plants with endogenous L-type inserts showed none of the distinct loci of GUS activity readily visible in material with no inserts, showing that L-type inserts could suppress not only the
uidA expression of genomic homologues, but also of copies added
in vitro.
Key words additive gene action - antisense RNA - biolistics - gene suppression - methylation