Effects of sucrose concentration on the accumulation of anthocyanins in grape (Vitis vinifera) cell suspension

Abstract

A cell suspension of Vitis vinifera L. cv. Gamay Frédaux var. teinturier composed of 50% pigmented cells was grown in Gamborg B5 medium supplemented with (per litre) 250 mg casein hydrolysate, 0.1 mg α-naphthalenacetic acid, 0.2 mg kinetin, and either 20, 30, 50, or 60 g sucrose. In the presence of 20 and 30 g sucrose/L, growth of cells was characterized by a typical sigmoid pattern and maximum cell density was obtained in 30 g sucrose/L. In both media, the anthocyanin content of pigmented cells did not change significantly throughout the growth cycle. In the presence of 50 g sucrose/L, cell growth was characterized by an extended lag phase. During this period, there was a significant (P < 0.05) accumulation of anthocyanins in the pigmented cells. Browning of the medium and premature cell death was observed in medium with 60 g sucrose/L. Microscopic observation of the suspension revealed the occurrence of anthocyanoplasts, especially in anthocyanin-containing cells cultured in 50 g sucrose/L. In some instances anthocyanoplasts were observed in cells with nonpigmented vacuoles, thus supporting the thesis according to which anthocyanoplasts would be the site of anthocyanin synthesis. Key words: anthocyanins, anthocyanoplasts, cell culture, metabolism, sucrose, Vitis vinifera.

Effects of high ammonium concentrations on growth and anthocyanin formation in grape (Vitis vinifera L.) cell suspension cultured in a production medium

“…In previous studies, we have shown that high sucrose and low nitrate concentrations had a positive effect on the intracellular accumulation of peonidin 3-glucoside and overall anthocyanins while inhibiting growth of cells [4,5]. The mode of action of high sucrose concentration on the cell culture was identified as an osmotic stress [2,3]. Elsewhere, several studies have shown that both the type and amount of nitrogen source affect not only growth but also the pattern and the yield of secondary metabolites formed.…”

“…Reducing sugars were analyzed by the Nelson method [14]. Total sugars (reducing sugars plus enzymatically hydrolysed sucrose) were estimated by the same method after sucrose was hydrolysed by an invertase [2].…”

Pigments in Grape

“…The primary source of cells for suspension cultures for the production of grape anthocyanins come from the two intensely pigmented teinturier cultivars, "Gamay Freaux" or "Bailey Alicante," in which high osmotic potential has been shown to enhance anthocyanin production [110,111]. Elevating the sucrose content in the cell culture medium can increase the external osmotic potential and lead to an increase in anthocyanin accumulation, presumably through a mechanism similar to the osmotic stress induced under water deficit.…”

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

“…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]. However, most of these reports have relied on absorbance measurement after pigment extraction, and this extraction method can only provide the average level of pigment accumulation.…”

“…Moreover, even in highly pigmented plant-cell cultures, a considerable number of non-pigmented white cells are usually observed. Several studies have examined the percentages of pigmented cells among total cells by counting them under a microscope [4][5][6]12] and reported the wide distribution of anthocyanin content in cultured cells. In these cases, the pigmented cells were identified irrespective of the amount of the accumulated pigment and the identification depended on the criteria of individual examiners.…”