Suppression of beta-1,3-glucanase transgene expression in homozygous plants.
A chimeric construct containing the Nicotiana plumbaginifolia beta-1,3-glucanase gn1 gene was introduced into Nicotiana tabacum SR1 to produce high levels of the enzyme constitutively. We determined that the GN1 protein represents a basic beta-1,3-glucanase isoform which accumulates into the vacuoles of the transgenic plants. Analysis of the progeny of the transgenic plant with the highest levels of gn1 expression revealed an unexpected phenomenon of gene suppression. Plants hemizygous for the T-DNA locus contained high levels of gn1 mRNA and exhibited a 14-fold higher beta-1,3-glucanase activity than untransformed plants. However, the expression of gn1 was completely suppressed in the homozygous plants: no corresponding mRNA or protein could be detected. This suppression mechanism occurs at a post-transcriptional level and is under developmental control. In addition, by generating haploid plants we found that this silencing phenomenon is not dependent on allelic interaction between T-DNA copies present at the same locus of homologous chromosomes, but rather is correlated with the transgene dose in the plant genome. We postulate that high doses of GN1 protein relative to the level(s) of other still unknown plant products could trigger the cellular processes directed to suppress gn1 expression.
de Carvalho
F
F
Laboratorium voor Genetica, Universiteit Gent, Belgium.
Gheysen
G
G
Kushnir
S
S
Van Montagu
M
M
Inzé
D
D
Castresana
C
C
D015152:Blotting, Northern; D015180:Electrophoresis, Gel, Two-Dimensional; D015971:Gene Expression Regulation, Enzymologic; D004693:Glucan Endo-1,3-beta-D-Glucosidase / Q000235:genetics* / Q000378:metabolism; D006720:Homozygote; D007150:Immunohistochemistry; D030821:Plants, Genetically Modified; D010947:Plants, Toxic; D012313:RNA / Q000378:metabolism; D012323:RNA Processing, Post-Transcriptional; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics* / Q000254:growth & development; D014158:Transcription, Genetic
D011993:Recombinant Fusion Proteins; D012313:RNA; D004693:Glucan Endo-1,3-beta-D-Glucosidase