Induction of anthocyanin synthesis in relation to embryogenesis in a carrot suspension culture: Correlation of metabolic differentiation with morphological differentiation

Abstract

A system in which anthocyanin synthesis could be induced under a defined condition, was established in a carrot suspension culture. A cell suspension culture of carrot (Daucus carota L. cv. Kurodagosun) was subcultured for more than a year in a medium containing 5 × 10−7M 2,4‐dichlorophenoxyacetic acid (2,4‐D). At every subculture the cultures were sieved through nylon screens and the cells and cell clusters collected in the size range of 31–81 μm were transferred to a fresh medium. When the cells were transferred to a medium without auxin, synthesis of anthocyanin was induced. Zeatin promoted anthocyanin synthesis in a medium lacking auxin, with maximum yields of anthocyanin obtained at 10−7 to 10−8M zeatin, 2,4‐D at higher concentrations than 10−7M inhibited anthocyanin synthesis completely. The sieved cells were fractionated by Ficoll density gradient centrifugation. Somatic embryos were formed in the fraction of higher density (>14% of Ficoll) in a medium containing 10−7M zeatin but lacking auxin, while synthesis of anthocyanin was hardly observed. On the other hand, cells in the fraction of lower density (<12% of Ficoll) synthesized anthocyanin in the same medium, but formed few embryos. Forty to fifty percent of the total cells in this lighter cell fraction synthesized anthocyanin at a maximum. The similarity between anthocyanin synthesis and embryogenesis was observed in the time course as well as in the effects of growth regulators. The correlation between metabolic and morphological differentiation is discussed.

Production of Anthocyanins, Carotenoids, and Proanthocyanidins by Cultured Cells of Rabbiteye Blueberry (Vaccinium asheiReade)

“…33 ) In case of Vitis vinifera cell suspension culture, the effect of 2,4-D on anthocyanin formation depended on kinetin concentration, and also vice versa. 23) Sucrose at 6% stimulated anthocyanin formation, but it inhibited cell growth considerably (Table II).…”

Role of miniature inverted repeat transposable elements inserted into the promoter region of a carrot phenylalanine ammonia-lyase gene and its gene expression

“…Kurodagosun) cells subcultured in medium containing 2,4-D under undifferentiated conditions, without anthocyanin synthesis, were maintained as described previously (Ozeki and Komamine 1981). The dedifferentiated cells were cultured and harvested five days after their transfer to medium containing 2,4-D.…”

“…The carrot cells in the suspension cultures grow in medium containing 2,4-dichlorophenoxyacetic acid (2,4-D). Anthocyanin synthesis is induced when the cells are cultured in medium lacking 2,4-D (Ozeki and Komamine 1981). We studied the regulatory mechanism of the phenylalanime ammonia-lyase genes (PAL) in this system, because PAL is a well-known key enzyme at the branching point from primary metabolism to phenylpropanoid metabolism (Dixon and Paiva 1995;Weisshaar and Jenkins 1998).…”

“…We established a system of carrot suspension cell cultures in which anthocyanin synthesis is induced as "metabolic differentiation" (Ozeki and Komamine 1981). The carrot cells in the suspension cultures grow in medium containing 2,4-dichlorophenoxyacetic acid (2,4-D).…”

Isolation of Daucus carota ethylene insensitive3-like (DcEIL) involved in stress-inducible DcMYB1 expression in suspension-cultured carrot cells

“…Kurodagosun, were grown in a modified Lin and Staba (LS) medium containing 0.5 mM 2,4-dichlorophenoxyacetic acid (2,4-D) at 27°C, and subcultured every 7 days (Ozeki and Komamine 1981). cDNA-encoding proteins that bound to the Ϫ135 to Ϫ105 sequence of the DcMYB1 promoter were isolated using a yeast one-hybrid system.…”