Formation of Anthocyanins from Cells Cultured from Different Parts of Strawberry Plants

Abstract

Anthocyanin formation and structural identification were investigated in the cultured strawberry, Frugaria unannssa cv Shikinari. Friable callus tissues were obtained from apical meristems, leafs, and petioles grown on LS agar medium with 2,4-dichlorophenoxyacetic acid and benzyladenine. Using these calli, effects of auxins and cell inoculum size on pigment production were studied in suspension culture. Anthocyanin was produced under 8000 lux after preculture for 3 wk under 800 Iux in suspension culture. The main extracted pigment was identified as a peonidin-3-glucoside and the second major pigment was a cyanidin-3-glucoside by FABMS and NMR. Anthocyanin production was stimulated by 2+dichlorophenoxyacetic acid and its yield was about three times that of indole-3-acetic acid. An inoculum rate of 2g (fresh weight)/100 mL LS medium resulted in highest pigment production. The callus derived from the leaf was much higher in anthocyanin production than the others tissues of strawberry.

Quantitative determination of cultured strawberry-cell heterogeneity by image analysis: effects of medium modification on anthocyanin accumulation

“…Shikinari) were derived from strawberry plant leaves [9]. The strawberry cells were grown in a liquid Linsmaier-Skoog medium [18] supplemented with 30 g sucrose, 1 mg 2,4-dichlorophenoxyacetic acid (2,4-D), and 0.1 mg benzylaminopurine (BAP) per 1 l medium.…”

“…The medium pH was adjusted to 5.7 before autoclaving. Subculturing was carried out every week by inoculating 100 ml of fresh medium with 2 g-fresh cell weight (fcw) of inoculum cells [9]. The flasks were maintained at 26 • C on a reciprocating shaker at 100 strokes/min.…”

“…Absorbance of the extract solution was determined by a spectrophotometer at 530 nm. The average anthocyanin content was calculated based on the extinction coefficient (E 1% 1cm =680 at 530 nm), which was obtained using purified peonidin-3-glucoside from cultured strawberry cells as a standard [9].…”

“…Anthocyanin, one of the secondary metabolites of plant cells, is well known as a natural pigment for food colorants with low toxicity. Anthocyanins are produced by various plant-cell cultures, including wild carrot [1,2], sweet potato [3], grape [4][5][6][7][8] and strawberry [9][10][11][12][13] cultures. It has been well documented that anthocyanin accumulation in plant cell cultures is strongly influenced by culture conditions, including light intensity [4,12,[14][15][16], plant growth regulators [10,11], medium components [4,12,17] and sugar concentration [2,[5][6][7][8]11,12,14].…”

Enhanced anthocyanin production by repeated-batch culture of strawberry cells with medium shift

“…Strawberry cell suspension culture has become established for the production of anthocyanin and batch mode was adopted for all these cultures (Hong et al 1989;Mori et al 1993;Sato et al 1996;Zhang et al 1997Zhang et al , 1998Zhang et al , 1999. It seems a favorable and necessary undertaking to explore process optimization to enhance anthocyanin productivity in strawberry cell culture.…”