Activation tagging: a means of isolating genes implicated as playing a role in plant growth and development

Richard Walden, Klaus Fritze, Hiroaki Hayashi, Edvins Miklashevichs, Hinrich Harling, Jeff Schell

Research output: Contribution to journalArticlepeer-review

79 Citations (Scopus)

Abstract

Activation T-DNA tagging has been used to generate a variety of tobacco cell lines selected by their ability to grow either in the absence of auxin or cytokinin in the culture media, or under selective levels of an inhibitor of polyamine biosynthesis. The majority of the cell lines studied in detail contain single T-DNA inserts genetically co-segregating with the selected phenotype. While most of the plants regenerated from the mutant cell lines appear phenotypically normal, several display phenotypes which could be inferred to result from disturbances in the content, or the metabolism, of auxins and cytokinins, or polyamines. The tagging vector is designed to allow the isolation of tagged plant genes by plasmid rescue. Confirmation that the genomic sequence responsible for the selected phenotype has indeed isolated is provided by PEG-mediated protoplast DNA uptake of rescued plasmids followed by selection for protoplast growth under the original selective conditions. Several plasmids have been rescued from the mutant lines which confer on transfected protoplasts the ability to grow either in the absence of auxin or cytokinin in the culture media, or under selective levels of an inhibitor of polyamine biosynthesis. This review describes the background to activation tagging and our progress in characterizing the genes that have been tagged in the mutant lines we have generated.

Original languageEnglish
Pages (from-to)1521-1528
Number of pages8
JournalPlant Molecular Biology
Volume26
Issue number5
DOIs
Publication statusPublished - Dec 1994

Keywords

  • activation tagging
  • Arabidopsis
  • auxin
  • cytokinin
  • polyamines
  • signal transduction
  • T-DNA tagging

Field of Science

  • 1.6 Biological sciences
  • 3.1 Basic medicine
  • 3.2 Clinical medicine

Publication Type

  • 1.1. Scientific article indexed in Web of Science and/or Scopus database

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