Comparability of imazapyr-resistant Arabidopsis created by transgenesis and mutagenesis

Transgenic Res. 2012 Dec;21(6):1255-64. doi: 10.1007/s11248-012-9597-z. Epub 2012 Mar 21.

Abstract

The Arabidopsis CSR1 gene codes for the enzyme acetohydroxyacid synthase (AHAS, EC 2.2.1.6), also known as acetolactate synthase, which catalyzes the first step in branched-chain amino acid biosynthesis. It is inhibited by several classes of herbicides, including the imidazolinone herbicides, such as imazapyr; however, a substitution mutation in csr1-2 (Ser-653-Asn) confers selective resistance to the imidazolinones. The transcriptome of csr1-2 seedlings grown in the presence of imazapyr has been shown in a previous study (Manabe in Plant Cell Physiol 48:1340-1358, 2007) to be identical to that of wild-type seedlings indicating that AHAS is the sole target of imazapyr and that the mutation is not associated with pleiotropic effects detectable by transcriptome analysis. In this study, a lethal null mutant, csr1-7, created by a T-DNA insertion into the CSR1 gene was complemented with a randomly-inserted 35S/CSR1-2/NOS transgene in a subsequent genetic transformation event. A comparison of the csr1-2 substitution mutant with the transgenic lines revealed that all were resistant to imazapyr; however, the transgenic lines yielded significantly higher levels of resistance and greater biomass accumulation in the presence of imazapyr. Microarray analysis revealed few differences in their transcriptomes. The most notable was a sevenfold to tenfold elevation in the CSR1-2 transcript level. The data indicate that transgenesis did not create significant unintended pleiotropic effects on gene expression and that the mutant and transgenic lines were highly similar, except for the level of herbicide resistance.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / drug effects
  • Arabidopsis / genetics*
  • Arabidopsis / metabolism
  • Arabidopsis Proteins / genetics*
  • Arabidopsis Proteins / metabolism
  • Gene Expression Profiling*
  • Gene Expression Regulation, Plant
  • Genes, Plant*
  • Genetic Complementation Test
  • Herbicide Resistance / genetics*
  • Imidazoles / pharmacology*
  • Mutagenesis
  • Mutation / genetics
  • Niacin / analogs & derivatives*
  • Niacin / pharmacology
  • Oligonucleotide Array Sequence Analysis
  • Plants, Genetically Modified / drug effects
  • Plants, Genetically Modified / genetics*
  • Plants, Genetically Modified / metabolism

Substances

  • Arabidopsis Proteins
  • Imidazoles
  • Niacin
  • imazapyr