Enhancement of anthocyanin production from grape (Vitis vinifera) callus in a viscous additive-supplemented medium

N-medium, which is the medium supplemented with carboxymethyl cellulose (CMC) as a viscous additive, was applied to the anthocyanin production from grape callus, Vitis vinifera cv. Bailey alicant A. Although the cell growth decreased with increasing CMC content in the medium, the anthocyanin content per packed cell volume (PCV) increased significantly. When N-B5-medium (0.8%), B5-medium containing 0.8% CMC, was used, approximately three times higher level of anthocyanin production was obtained than that from the control culture without CMC addition. In the repeated batch culture using N-B5-medium (0.8%), the productivity was found to continue at a high level during 30 batch cultures, while no anthocyanin productivity was obtained after 16th batch in the control culture. When the completely white callus after 16th batch culture was transferred to N-B5-medium (0.8%) and cultured repeatedly, anthocyanin productivity was completely recovered after 16th batch culture in N-B5-medium. In contrast, when calli obtained from N-B5-medium (0.8%) was cultured in conventional medium, the productivity was gradually decreased and became almost 0 after 8th repeated batches. Size distribution of calli obtained from the culture in N-B5-medium (0.8%) was also investigated. After repeated batch culture in N-B5-medium (0.8%), the size of calli was shifted to large size and the fraction of calli with less than 0.2 mm decreased remarkably. AM calli in the fraction of more than 0.6 mm in size contained pigmented purple cells, while in the fraction of less than 0.2 mm in size the ratio of calli containing pigmented purple cells was less than 50%. From these results, it was confirmed that the enhanced and prolonged production of anthocyanin was due to the shift of size distribution to larger size and it was suggested that the shift of cell size was caused by lowering the damage to grape callus by hydrodynamic stress via increase of medium viscosity. (C) 2001 Elsevier Science B.V. All rights reserved.