Systematic reverse genetics of transfer-DNA-tagged lines of arabidopsis: Isolation of mutations in the cytochrome P450 gene superfamily

Rodney G. Winkler, Michael R. Frank, David W. Galbraith, René Feyereisen, Kenneth A. Feldmann

Research output: Contribution to journalArticle

85 Scopus citations

Abstract

We have developed an efficient reverse-genetics protocol that uses expedient pooling and hybridization strategies to identify individual transfer-DNA insertion lines from a collection of 6000 independently transformed lines in as few as 36 polymerase chain reactions. We have used this protocol to systematically isolate Arabidopsis lines containing insertional mutations in individual cytochrome P450 genes. In higher plants P450 genes encode enzymes that perform an exceptionally wide range of functions, including the biosynthesis of primary metabolites necessary for normal growth and development, the biosynthesis of secondary products, and the catabolism of xenobiotics. Despite their importance, progress in assigning enzymatic function to individual P450 gene products has been slow. Here we report the isolation of the first 12 such lines, including one (CYP83B1-1) that displays a runt phenotype (small plants with hooked leaves), and three insertions in abundantly expressed genes. The DNAs used in this study are publicly available and can be used to systematically isolate mutants in Arabidopsis.

Original languageEnglish (US)
Pages (from-to)743-750
Number of pages8
JournalPlant physiology
Volume118
Issue number3
DOIs
StatePublished - Jan 1 1998

ASJC Scopus subject areas

  • Physiology
  • Genetics
  • Plant Science

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