Simultaneous suppression of three genes related to brassinosteroid (BR) biosynthesis altered campesterol and BR contents, and led to a dwarf phenotype in Arabidopsis thaliana

Plant Cell Rep. 2010 Apr;29(4):397-402. doi: 10.1007/s00299-010-0830-z. Epub 2010 Feb 19.

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

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / enzymology
  • Arabidopsis / genetics*
  • Arabidopsis / growth & development
  • Arabidopsis Proteins / genetics*
  • Cholesterol / analogs & derivatives*
  • Cholesterol / biosynthesis
  • Cytochrome P-450 Enzyme System / genetics
  • DNA, Complementary / genetics
  • Gene Expression Regulation, Plant
  • Gene Knockdown Techniques
  • Genes, Plant
  • Methyltransferases / genetics
  • Phenotype
  • Phytosterols / biosynthesis*
  • Plant Growth Regulators / biosynthesis*
  • Plants, Genetically Modified / enzymology
  • Plants, Genetically Modified / genetics
  • Plants, Genetically Modified / growth & development
  • RNA Interference
  • RNA, Double-Stranded / genetics
  • RNA, Plant / genetics
  • Reverse Transcriptase Polymerase Chain Reaction
  • Steroids / biosynthesis*

Substances

  • AT3G50660 protein, Arabidopsis
  • Arabidopsis Proteins
  • DET2 protein, Arabidopsis
  • DNA, Complementary
  • Phytosterols
  • Plant Growth Regulators
  • RNA, Double-Stranded
  • RNA, Plant
  • Steroids
  • campesterol
  • Cytochrome P-450 Enzyme System
  • Cholesterol
  • Methyltransferases
  • sterol methyltransferase 2, Arabidopsis