Carrot (Daucus carota L.)

Methods Mol Biol. 2006:344:3-12. doi: 10.1385/1-59745-131-2:3.

Abstract

Plants are susceptible to infection by a broad range of fungal pathogens. Many horticulturally important crop species lack adequate genetic resistance to disease. Studies on potential mechanisms of disease resistance in plants have revealed the importance of a range of pathogenesis-related (PR) proteins with antifungal activity in reducing colonization of plant tissues by pathogens. We are evaluating a range of PR-proteins, through heterologous expression in transgenic carrot tissues, for their effects on fungal disease development. The protocols for carrot transformation with a thaumatin-like protein are described. In addition, the use of herbicide resistance as a selectable marker in carrot transformation is illustrated. In this protocol, petiole segments from carrot seedlings are exposed to Agrobacterium for 10-30 min and co-cultivated for 3 d, after which herbicide selection is imposed until embryogenic calli are produced after 8-12 wk. The transfer of the embryogenic calli to hormone-free medium yields transgenic plantlets. This genetic transformation protocol has supported the generation of transgenic carrot plants with defined T-DNA inserts at the rate of between 1 and 3 Southern positive independent events out of 100.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Agrobacterium tumefaciens / cytology
  • Agrobacterium tumefaciens / genetics*
  • Cell Culture Techniques
  • Culture Media
  • Daucus carota / genetics*
  • Daucus carota / metabolism
  • Daucus carota / microbiology
  • Fungi / physiology
  • Genetic Vectors
  • Immunity, Innate / genetics
  • Plant Diseases / microbiology
  • Plant Proteins / genetics
  • Plants, Genetically Modified / metabolism
  • Plants, Genetically Modified / microbiology
  • Polymerase Chain Reaction
  • Proteomics
  • Seedlings / genetics
  • Seedlings / metabolism
  • Seedlings / microbiology
  • Tissue Culture Techniques
  • Transformation, Genetic*

Substances

  • Culture Media
  • Plant Proteins