Homologous recombination in plant cells after Agrobacterium-mediated transformation

K Y Lee, P Lund, K Lowe, P Dunsmuir

Research output: Contribution to journalArticlepeer-review

Abstract

A single amino-acid change in the acetolactate synthase (ALS) protein of tobacco confers resistance to the herbicide chlorsulfuron. A deleted, nonfunctional fragment from the acetolactate synthase gene, carrying the mutant site specifying chlorsulfuron resistance plus a closely linked novel restriction site marker, was cloned into a binary vector. Tobacco protoplasts transformed with Agrobacterium tumefaciens carrying this vector yielded chlorsulfuron-resistant colonies. DNA gel blot analysis of DNA from these colonies suggested that in three transformants homologous recombination had occurred between the endogenous ALS gene and the deleted ALS gene present in the incoming T-DNA. Plants were regenerated from these chlorsulfuron-resistant colonies, and in two of the transformants, genetic analysis of their progeny showed that the novel gene segregated as a single Mendelian locus. Possible models for the generation of these recombinant plants are discussed.
Original languageEnglish
Pages (from-to)415-25
Number of pages11
JournalThe Plant Cell
Volume2
Issue number5
DOIs
Publication statusPublished - May 1990

Keywords

  • Acetolactate Synthase
  • Triazines
  • Drug Resistance
  • Herbicides
  • Plasmids
  • Agrobacterium tumefaciens
  • Plants, Toxic
  • Base Sequence
  • Transformation, Genetic
  • Recombination, Genetic
  • Molecular Sequence Data
  • Tobacco
  • Crosses, Genetic
  • Genes, Plant
  • Sulfonamides

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