An active role for endogenous beta-1,3-glucanase genes in transgene-mediated co-suppression in tobacco.
Post-transcriptional gene silencing (PTGS) is characterized by the accumulation of short interfering RNAs that are proposed to mediate sequence-specific degradation of cognate and secondary target mRNAs. In plants, it is unclear to what extent endogenous genes contribute to this process. Here, we address the role of the endogenous target genes in transgene-mediated PTGS of beta-1,3-glucanases in tobacco. We found that mRNA sequences of the endogenous glucanase glb gene with varying degrees of homology to the Nicotiana plumbaginifolia gn1 transgene are targeted by the silencing machinery, although less efficiently than corresponding transgene regions. Importantly, we show that endogene-specific nucleotides in the glb sequence provide specificity to the silencing process. Consistent with this finding, small sense and antisense 21- to 23-nucleotide RNAs homologous to the endogenous glb gene were detected. Combined, these data demonstrate that a co-suppressed endogenous glucan ase gene is involved in signal amplification and selection of homologous targets, and show that endogenous genes can actively participate in PTGS in plants. The findings are introduced as a further sophistication of the post-transciptional silencing model.
Sanders
Matthew
M
Department of Plant Systems Biology, Flanders Interuniversity Institute for Biotechnology (VIB), Ghent University, K.L. Ledeganckstraat 35, Belgium.
Maddelein
Wendy
W
Depicker
Anna
A
Van Montagu
Marc
M
Cornelissen
Marc
M
Jacobs
John
J
D001483:Base Sequence; D018744:DNA, Plant; D020868:Gene Silencing; D043326:Glucan 1,3-beta-Glucosidase; D008969:Molecular Sequence Data; D030821:Plants, Genetically Modified; D012333:RNA, Messenger / Q000235:genetics; D012689:Sequence Homology, Nucleic Acid; D014026:Nicotiana / Q000201:enzymology / Q000235:genetics*; D019076:Transgenes; D001617:beta-Glucosidase / Q000235:genetics*
D018744:DNA, Plant; D012333:RNA, Messenger; D001617:beta-Glucosidase; D043326:Glucan 1,3-beta-Glucosidase