(49e) Derivation of Non-Anthocyanin-Producing Grape Cell Culture from a Stable Anthocyanin-Producing Grape Cell Culture: an Example of Reverse Engineering

Production instability in plant cell culture is a major obstacle to commercial production of valuable secondary metabolites, such as, new drugs, or natural pigments. The production of anthocyanin in grape cell culture was used as a model to understand the physiological basis for instability of secondary metabolites production in plant cell culture. The study began with a stable anthocyanin-producing grape cell line. In order to compare the differences between anthocyanin-producing cell and non-anthocyanin-producing cell in physiological and genetic level, a stable non-anthocyanin-producing cell line with same genetic background was derived from the anthocyanin-producing grape cell line. The results show that reducing sucrose concentration to 10 g/l in solid B5 medium can help to significantly lower the anthocyanin production in grape callus and form white callus. In addition to the low sucrose level in the medium, restriction of gas exchange was a critical stress factor in the formation of non-anthocyanin-producing suspension cultures. The anthocyanin content was 0.1 mg/ml and 2.9 mg/ml in the non-anthocyanin-producing suspension culture and the original anthocyanin-producing suspension culture respectively. For enhancement of production of anthocyanin by plant cell cultures, many efforts in creating a more supportive environment such as increasing sucrose level, lights, oxygen supply have been studied and outstanding results have been reported. On the contrary, this research suggests that environmental stress, such as reducing the level of carbon source such as sucrose concentration and restriction of gas exchange, could be methods to derive a non-anthocyanin-producing cell line with same genetic background from a stable anthocyanin-producing cell line.