Temporal and Spatial Heterogeneity in the Accumulation of Anthocyanins in Cell Cultures ofCatharanthus roseus(L.) G.Don.

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Abstract
The accumulation of anthocyanins, a group of pigmented secondary metabolites, in cell cultures of the Madagascar periwinkle Catharanthus roseus has been investigated. In these cultures it was found that anthocyanin accumulation was restricted to the post-division phase of the culture growth cycle, during which the cultures became deep purple in colour. As a result of anthocyanin visibility it has been possible to ascertain that accumulation of these metabolites occurred in only a small proportion of the cell population. Approximately 10% of cells regularly accumulated detectable levels. Considerable variation within this ‘productive’ population was observed and using a standard integrating microdensitometer it has been possible to quantify directly this heterogeneity and compare it with data obtained from whole plants. Analysis has revealed that the variation in both intracellular anthocyanin content and concentration in cell cultures was much greater than that observed within tissues of mature plants. Significant differences in mean values were however found between the whole plant tissues. The relevance of this temporal and spatial heterogeneity observed in vitro to our understanding of the control of secondary metabolite accumulation and to the potential use of tissue culture systems as a means to produce these compounds is discussed.