An intron sense suppression construct targeting soybean FAD2-1 requires a double-stranded RNA-producing inverted repeat T-DNA insert.
We demonstrate that the transformation of soybean (Glycine max) with sense suppression constructs using intron sequences from the fatty acid oleyl Delta12 desaturase gene FAD2-1A leads to efficient and specific reduction of FAD2-1 transcripts in developing seeds, increased oleic acid, and decreased polyunsaturated fatty acids. The related FAD2-2 transcripts are only marginally affected. Despite screening a large number of independent transformants, no single-copy efficacious transformants could be found. Invariably, all the least complex transgenic loci have two T-DNA copies in an inverted repeat configuration, centered at the right borders. We show that this T-DNA configuration produces an inverted repeat transcript and that small interfering RNAs accumulate against the target sequence.
Mroczka
Andrew
A
Monsanto Company, Calgene Campus, Davis, CA 95616, USA.
Roberts
Peter D
PD
Fillatti
Joanne J
JJ
Wiggins
B Elizabeth
BE
Ulmasov
Tim
T
Voelker
Toni
T
D004269:DNA, Bacterial / Q000235:genetics; D018744:DNA, Plant / Q000235:genetics; D044943:Fatty Acid Desaturases / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D007438:Introns; D016254:Mutagenesis, Insertional; D010940:Plant Proteins / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics; D012330:RNA, Double-Stranded / Q000235:genetics*; D018749:RNA, Plant / Q000235:genetics; D034741:RNA, Small Interfering / Q000235:genetics; D012639:Seeds / Q000254:growth & development; D013025:Glycine max / Q000201:enzymology / Q000235:genetics*; D014170:Transformation, Genetic
D004269:DNA, Bacterial; D018744:DNA, Plant; D010940:Plant Proteins; D012330:RNA, Double-Stranded; D018749:RNA, Plant; D034741:RNA, Small Interfering; C036483:T-DNA; D044943:Fatty Acid Desaturases; C040448:delta-12 fatty acid desaturase