Transgenic Research

, Volume 21, Issue 6, pp 1255–1264 | Cite as

Comparability of imazapyr-resistant Arabidopsis created by transgenesis and mutagenesis

  • Jaimie Schnell
  • Hélène Labbé
  • Nik Kovinich
  • Yuzuki Manabe
  • Brian Miki
Original Paper

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.

Keywords

Mutagenesis Transgenesis Transcriptome Unintended effects Acetohydroxyacid synthase Acetolactate synthase 

Supplementary material

11248_2012_9597_MOESM1_ESM.jpg (753 kb)
Online Resource 1. Arabidopsis CSR1 mutant alleles and transgenes (JPG 753 kb)
11248_2012_9597_MOESM2_ESM.pdf (81 kb)
Online Resource 2. Genes up- or down-regulated greater than twofold (P < 0.05) in transgenic lines. M is log2 fold change and P is FDR-adjusted P-value for Student’s t-test (PDF 81 kb)

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Copyright information

© Her Majesty the Queen in Right of Canada 2012

Authors and Affiliations

  • Jaimie Schnell
    • 1
    • 2
  • Hélène Labbé
    • 1
  • Nik Kovinich
    • 1
  • Yuzuki Manabe
    • 1
    • 3
  • Brian Miki
    • 1
  1. 1.Bioproducts and Bioprocesses, Research BranchAgriculture and Agri-Food CanadaOttawaCanada
  2. 2.Plant and Biotechnology Risk Assessment UnitCanadian Food Inspection AgencyOttawaCanada
  3. 3.Feedstocks Division, Joint BioEnergy InstituteLawrence Berkely National LaboratoryBerkeleyUSA

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