Abstract
We generated transgenic lines of Arabidopsis thaliana with an RNA interference construct that expressed hairpin double-stranded RNA for DET2:DWF4:SMT2 to induce sequence-specific RNA silencing. In transgenic plants, expressions of DET2, DWF4, and SMT2 were simultaneously reduced, and the campesterol content was increased by up to 420% compared to the level in the wild-type plant. Triple knock-down of the DET2, DWF4, and SMT2 enzymes also resulted in reduction of brassinosteroid (BR)-specific biosynthesis intermediates. Transgenic plants harboring the RNA interference construct displayed a semi-dwarf phenotype due to altered development. Our findings indicate that redesigning of plant architecture is possible through simultaneous suppression of multiple genes involved in BR biosynthesis.
Publication types
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Research Support, Non-U.S. Gov't
MeSH terms
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Arabidopsis / enzymology
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Arabidopsis / genetics*
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Arabidopsis / growth & development
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Arabidopsis Proteins / genetics*
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Cholesterol / analogs & derivatives*
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Cholesterol / biosynthesis
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Cytochrome P-450 Enzyme System / genetics
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DNA, Complementary / genetics
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Gene Expression Regulation, Plant
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Gene Knockdown Techniques
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Genes, Plant
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Methyltransferases / genetics
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Phenotype
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Phytosterols / biosynthesis*
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Plant Growth Regulators / biosynthesis*
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Plants, Genetically Modified / enzymology
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Plants, Genetically Modified / genetics
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Plants, Genetically Modified / growth & development
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RNA Interference
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RNA, Double-Stranded / genetics
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RNA, Plant / genetics
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Reverse Transcriptase Polymerase Chain Reaction
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Steroids / biosynthesis*
Substances
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AT3G50660 protein, Arabidopsis
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Arabidopsis Proteins
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DET2 protein, Arabidopsis
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DNA, Complementary
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Phytosterols
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Plant Growth Regulators
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RNA, Double-Stranded
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RNA, Plant
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Steroids
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campesterol
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Cytochrome P-450 Enzyme System
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Cholesterol
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Methyltransferases
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sterol methyltransferase 2, Arabidopsis