Initiation and Establishment of Cell Suspension Cultures of Carrot (Daucus carota)

Abstract

The culture of plant tissues in an agitated liquid medium eliminates many of the disadvantages ascribed to the culture of tissues on agar. Movement of the tissue in relation to the nutrient medium facilitates gaseous exchange, removes any polarity of the tissue due to gravity, and eliminates nutrient gradients within the medium and at the surface of the cells. The incubation of a friable callus in a liquid nutrient medium agitated on a shaking machine, will eventually give a suspension of cells which are more amenable to experimental manipulation than callus grown on agar. The technique described here is one in which a callus of Daucus carota is suspended in a liquid nutrient medium and is subjected to a series of manipulations to ensure the production of an actively growing cell suspension consisting of single cells and very small aggregates of 2–15 cells.