JPH0636745B2 - Method for producing red pigment by cultured statice cells - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION [Industrial field of use] The present invention relates to a method for producing a red pigment by a statice culture cell.

[Conventional technology]

In recent years, it has been desired to use natural dyes, which are safer than synthetic dyes, as food coloring agents. Natural pigments are produced by extracting them from plants, etc., but they require vast land, time, and labor for cultivation.
It is difficult to obtain a stable quality product depending on the soil.

Conventionally, a method for producing a red pigment from plants such as safflower cultured cells is known, but a method for producing a red pigment using Larchium sp. (Limonium Latifolium O. Kuntze) is not yet known. .

[Problems to be Solved by the Invention]

An object of the present invention is to provide a method for efficiently producing a stable and constant quality red pigment from cultured cells of leaves or stems of statice.

[Means for Solving the Problems]

TECHNICAL FIELD The present invention relates to a method for producing a red pigment by cultured statice cells, which comprises culturing cells of leaves or stems of statice under white light using an agar or liquid medium containing auxin and cytocanin.

The statice used in the present invention is a halophyte belonging to the genus Pinus, and is usually cultivated as a horticultural species, and is one year such as limonium sinuatum (L.sinuatum Mill) or limonium suororowii (L.suworowii O.Kuntze). Grass, limonium latifolium (L.latifolium OK
untze) and limonium tartaricum (L.tataricum Mil
Perennial plants such as l.) are used.

As the statice cells used for callus induction, statice leaf or stem tissues are used. These tissues were placed on agar or liquid medium containing auxin and cytokinin and cultured while irradiating with white light of, for example, 2000 ux to induce callus, and then passaged, for example, every 30 days under the same culture conditions as above. Red pigment is produced by repeating. As it continues to passage, it changes from bleached to black and then from black to red.

The auxin used in the present invention includes indole-
3-acetic acid (1AA), 1-naphthalene acetic acid (NAA),
2,4-dichlorophenoxyacetic acid (2,4-D), 2,
4,5-Trichlorophenoxyacetic acid (2,4,5-T)
And the like, and as cytocanin, zeatin,
Examples include dihydrozeatin, ribosylzeatin, 6-benzyladenine (BA), kinetin and the like.

The medium components used in the present invention are not particularly limited, and known Murashige-Skoog medium (1962), Linsmaier-Skoog
Medium (1965), White medium (1963), GamborgB-
5 (1968), Nitsch medium (1951), Heller medium (1953), Schenk-Hildebrandt medium (1972),
Nitsch-Nitsch medium (1967), Kohlenbach-Schmidt
A medium (1975), a Knop medium (1865), or a modified medium obtained by adding sucrose, glucose, raffinose, fructose, pentose, maltose or the like as a carbon source is used. In the case of agar medium, agar may be added to these media at about 0.7 to 1.5%.

In the present invention, in order to efficiently produce the red pigment by the statice cultured cells, 100 to 20,000 is required.
It is necessary to irradiate white light of ux, preferably about 2000 ux. With white light outside this range, the productivity of the red dye will be poor. As the white light, for example, light from a fluorescent lamp, a white light bulb, or the like can be used. The culture temperature is not particularly limited, but is preferably around 25 ° C. In the case of a liquid medium, shaking or rotary culture may be performed.

〔Example〕

 Hereinafter, the present invention will be described in detail with reference to Examples.

Example 1 Stirchis leaves and stems were cut, sterilized, and then pH 6.
0, kinetin ^{10 -5 M, NAA10 -5 M,} Murashige-Skoog containing 3% sucrose and 1% agar
(Hereinafter, abbreviated as “MS”)
The callus was induced by culturing at 25 ° C. while irradiating white light of x (light source: fluorescent lamp).

The obtained callus of statice is shown in Table 1 in various media (added hormone: kinetin 10 ⁻⁵ M, NAA10.
^-5 M), and the amount of proliferation after 30 days was examined. First
The table shows the amount of proliferation of 1 g of cells after 30 days.
There was no difference in growth depending on the medium except for p medium.

Therefore, using the MS medium used for callus induction, the culture was continued while subcultured every 30 days under the same culture conditions as the above callus induction. The callus was decolorized at the 4th passage, turned black at the 6th passage, and red pigment was observed at the 9th passage. The pigment was extracted from the cultured callus. Extraction is 1g
The cells were added with 10 m of methanol containing 0.5% of hydrochloric acid and allowed to stand at room temperature for 24 hours.
The extract had a maximum absorption at 530 nm in the visible region. Measuring the absorbance of the extract at 530 nm,
The dye amount was calculated, and the results are shown in Table 2.

Comparative Example 1 Callus was induced in the same manner as in Example 1 except that 2000 ux of white light was not used and culture was performed in the dark, and culture was performed while subculturing, and pigment was extracted and extracted. The dye amount of the liquid was calculated. The results are shown in Table 2.

Comparative Example 2 Callus was induced in the same manner as in Example 1 except that the medium in which kinetin and NAA were not added was used, and the cells were cultured while being passaged. The pigment was extracted to extract the pigment amount of the extract. Was calculated. The results are shown in Table 2.

Comparative Example 3 Callus was induced in the same manner as in Example 1 except that the medium in which kinetin and NAA were not added was used, and the darkening was further performed in the dark.
The pigment was extracted and the amount of pigment in the extract was calculated. Second result
Shown in the table.

From Table 2, in this example in which the hormone was added and cultured by irradiating with light, about 30 times more pigment amount was accumulated than in the case of not irradiating with light and not adding the hormone (Comparative Example 3). In addition, the amount of pigment was accumulated about 7 times as much as that obtained by culturing in the dark with the addition of hormones (Comparative Example 1). Indicates that about twice the amount of dye is accumulated.

〔The invention's effect〕

According to the production method of the present invention, a red pigment can be stably produced by culturing statice cells in a predetermined medium under white light and subculturing.

Claims (1)

[Claims] 1. A method for producing a red pigment by cultured statice cells, which comprises culturing cells of leaves or stems of statice under a white light using an agar or liquid medium containing auxin and cytocanin.