Increase in anthocyanin yield from wild-carrot cell cultures by a selection system based on cell-aggregate size

Abstract

Wild-carrot (Daucus carota L.) cell cultures were screened to yield small (less than 63 μm) or large (greater than 170 μm) cell aggregates which were then subcultured. Cultures of the small-size class had a higher, those of the large-size class a lower anthocyanin yield than the unscreened culture. This relationship became accentuated with an increasing number of passages with screening prior to subculture. At the end of six months (12 passages), the pigment yield of the small-size class was triple that of the unscreened cells. Following this selection period, the tendency of the small-size fraction to increase in clump size when subcultured without screening was much less than that of freshly isolated cell aggregates of the same size. These observations may be explainable on the basis of a differential distribution of cytokinin between aggregates of different sizes. High levels of cytokinin inhibit anthocyanin accumulation and inhibit cell separation; these effects result in large cell aggregates having low levels of anthocyanin. In support of this hypothesis, it is shown that addition of kinetin to cultures of small cell aggregates causes an increase in the size of cell aggregates and a parallel decrease in anthocyanin yield.

Isolation and Identification of the Phenols of Paul's Scarlet Rose Stems and Stem-Derived Suspension Cultures

“…Thus, our results are in direct contrast to those of Kinnersely and Dougall (13) and Ozeki and Komamine (20) where sieving significantly enhanced the anthocyanin content of carrot suspension cultures. In their research with carrot cultures, Kinnersely and Dougall (13) have speculated that small cell aggregates produce more anthocyanins because they contain lower endogenous levels ofcytokinins, which inhibit anthocyanin synthesis. If this is true, then the failure of rose cells to behave in a similar manner suggests that the endogenous cytokinin levels of sieved rose cells is still high enough to inhibit increased phenolic synthesis.…”

“…These results are consistent with the common report that most cultured plant cells have lower levels of secondary products than the plant organs from which they were started and that the levels of these compounds in cultured plant cells tends to diminish with increasing numbers of transfers (8). ( 13). It has been shown that the ability of carrot cell cultures to accumulate anthocyanin is reversible (9) and that a positive correlation exists between decreased clump size and anthocyanin production ( 13,20 and a second subline for five transfer periods), and two by sieving through 500-,gm screens (one subline for four subculturing periods and another subline for 12 subculturing periods).…”

“…( 13). It has been shown that the ability of carrot cell cultures to accumulate anthocyanin is reversible (9) and that a positive correlation exists between decreased clump size and anthocyanin production ( 13,20 and a second subline for five transfer periods), and two by sieving through 500-,gm screens (one subline for four subculturing periods and another subline for 12 subculturing periods). Sieving failed to enhance, in either a quantitative (Fig.…”

Nutrient utilization during biomass and anthocyanin accumulation in suspension cultures of wild carrot cells

“…The data on nutrient levels in the culture medium of carrot cell suspension cultures compliment that for suspension culture of Catharanthus roseus [1,18], tobacco [15,17], rice [4], Anchusa officinalis [5], Coptisjaponica [21], and Ipomoea cultures [23].…”

“…For experiments, subclones were grown in anthocyanin production medium (WCM-IMP) [18], modified by doubling the phosphate and ammonium concentrations and reducing the sugar concentration in an attempt to achieve carbohydrate limitation of growth. Cultures were grown in the dark in 50-ml batches of medium in 500-ml conical flasks with silicone sponge closures (Bellco, Inc.), agitated at 125 rpm.…”