Inheritance and expression of a transgene insert in an aneuploid tobacco line.
A T-DNA locus comprising nptII, uidA and nos genes--all under the control of the nos promoter (this locus was designated K because it encodes resistance to Kanamycin)--was found to be inherited erratically in a transgenic tobacco line. This anomalous behavior was partially explained following a karyotype analysis of plants representing several generations: these plants were aneuploids, presumably for the K-containing chromosome. During four generations of sexual propagation, transgenic plants that were either trisomic or tetrasomic for the K-containing chromosome (i.e. 2n = 49 or 2n = 50, respectively) were obtained. The trisomic plants (2n = 48 + 1) were virtually indistinguishable phenotypically from normal euploids (2n = 4x = 48), whereas the tetrasomic plants (2n = 48 + 2) were smaller, had somewhat misshapen leaves and exhibited reduced fertility. Although the amount of NPTII protein in different trisomic (K--, KK-, KKK) and tetrasomic (KK--, KKK-) plants was generally consistent with a K dosage effect, the genetic behavior of each trisomic--with respect to segregation of KanR and marker gene activity in progeny--was unique and not completely explicable by invoking aneuploidy. Specifically, unexpected gains or losses of K could occur, suggesting the formation of double reductional gametes and/or frequent gene conversion at this locus. The susceptibility of K locus marker genes to trans-inactivation in the trisomic and tetrasomic lines was tested by crossing in partially homologous silencing loci. In all transgenotypes tested, the three K marker genes were sensitive to trans-silencing, which was accompanied by methylation in all copies of the nos promotor. In addition to this directed inactivation/methylation, the K locus could also undergo infrequent, spontaneous partial methylation, which produced stable epialleles. In most plants, however, the multiple copies of the nos promoter at this locus remained unmethylated and active through four generations in all transgenotypes examined. The significance of these results for irregular inheritance patterns, aneuploid syndromes and homology-dependent gene silencing is discussed.
Matzke
M A
MA
Institute of Molecular Biology, Austrian Academy of Sciences, Salzburg.
Moscone
E A
EA
Park
Y D
YD
Papp
I
I
Oberkofler
H
H
Neuhuber
F
F
Matzke
A J
AJ
D000782:Aneuploidy; D018744:DNA, Plant / Q000378:metabolism*; D005785:Gene Conversion; D018506:Gene Expression Regulation, Plant; D005819:Genetic Markers; D007613:Kanamycin Resistance / Q000235:genetics; D008745:Methylation; D030821:Plants, Genetically Modified / Q000235:genetics*; D010947:Plants, Toxic; D012689:Sequence Homology, Nucleic Acid; D014026:Nicotiana / Q000235:genetics*
D018744:DNA, Plant; D005819:Genetic Markers