Differential gene expression between pigmented and non-pigmented cell culture lines of Daucus carota

Abstract

RNA differential display was used to look at differential transcript abundance between cell culture lines of wild carrot (Daucus carota L.) that either produce no pigments, or carotenoids only, or carotenoids and cyanidin-based anthocyanins. Nineteen partial cDNA clones were isolated and used in northern RNA analysis, confirming that seven of the corresponding genes were preferentially expressed in particular cell lines. For six of the seven differential clones, longer cDNAs were isolated from a cDNA library. Nucleotide sequence analysis allowed putative identification of the encoded products of three. The isolation of the cDNAs allowed the temporal variation in transcript abundance to be examined. For two out of five cDNAs tested the differential expression pattern was lost after a period of 24 months, representing approximately 52 subcultures. The results demonstrate the relevance of a differential RNA approach to trying to understanding the nature of mechanisms causing variability in plant cell cultures, the first step in bringing it under control for the improvement of culture technologies.