Proc Natl Acad Sci U S A 1993 Nov 1;90(21):10370-4

 

The maize transposable element system Ac/Ds as a mutagen in Arabidopsis: identification of an albino mutation induced by Ds insertion.

 

Long D, Martin M, Sundberg E, Swinburne J, Puangsomlee P, Coupland G

Molecular Genetics Department, Cambridge Laboratory, John Innes Centre, Norwich, United Kingdom.

A two-component transposon system based on the Ac element of maize was used as a mutagen in Arabidopsis thaliana. Transposition of a Ds element marked with a hygromycin-resistance gene was activated from four different locations in the Arabidopsis genome. The progeny of 201 plants carrying independent transposition events were screened for mutants with severe, visible phenotypes. Seven mutants were identified and four of them were analyzed genetically. Three mutations were shown to be very closely linked to a transposed copy of the element. Moreover, a mutation (alb3) causing an albino phenotype was conclusively shown to be caused by insertion of the Ds element: somatic and germinal reversion of the mutation occurred in the presence of the transposase gene but not in its absence, and in three revertants the Ds had excised from its position in the mutant line. The DNA adjacent to Ds in the mutant was isolated and it was demonstrated that revertants retained part of the 8-bp duplication caused by insertion of Ds. These experiments indicate that the Ac/Ds system can be used as an insertional mutagen in the heterologous host Arabidopsis, which will permit the isolation of genes from this species by transposon tagging.

 

MeSH Terms:

  • Arabidopsis/genetics*
  • Base Sequence
  • Corn/genetics*
  • DNA/isolation & purification
  • DNA Primers
  • DNA Transposable Elements*
  • Glucuronidase/metabolism
  • Glucuronidase/analysis
  • Molecular Sequence Data
  • Mutagenesis, Insertional*
  • Phenotype
  • Polymerase Chain Reaction/methods
  • Support, Non-U.S. Gov't

Substances:

  • DNA
  • DNA Transposable Elements
  • DNA Primers
  • Glucuronidase

PMID: 8234300, UI: 94052164