JP3074804B2 - Tissue culture method of Saffron Mesbe - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for effectively cultivating a pigment which is one of the active ingredients in edible and medicinal saffron.

[0002]

2. Description of the Related Art Saffron is a perennial herb of the Iridaceae family, Crocus sativas L .. (Crocus Sativus L.), whose stigma is dried is used as a crude drug. It is produced in Hiroshima, Kagawa, Oita and Iwate prefectures in Spain, France, Italy and Japan. It contains about 2% of carotenoid pigment, crocin (glycoside of crocetin), and bitter glycoside Picrocrocin.
Contains about 2% and 0.4-1.3% of essential oils. The main component of the essential oil is safranal (fragrance component peculiar to crude drugs).

[0003] This saffron is used as a spice, a food coloring, or as a medicinal product. However, since it uses a dried mesibet, it cannot be obtained in a natural product.
The unit price is also very high. In addition, natural products are difficult to obtain products of a certain quality due to changes in the climate. Therefore, various attempts have been made for tissue culture.

For example, Japanese Patent Application Laid-Open No. 62-275617 discloses Crocus Satibas L. That is, a method is disclosed in which a saffron pistil or an organ corresponding thereto is grown and hypertrophically matured using cytokinin or an artificial culture medium containing cytokinin and auxin. Cytokinin has been known for a long time as a substance that induces cell division of cultured plant cells, and it is also known that callus induction and cell growth are promoted when used in combination with auxin.

[0005] JP-A-63-109788 discloses Crocus Sativas L.A. Of the female organ of the present invention, cultured in an artificial culture medium containing cytokinin, and selected from the group consisting of safranal, picrocrosin and their analogs, crocin, crocetin and their analogs
It produces the above substances. However, it has long been known that saffron pistils contain these substances.

JP-A-63-160580 discloses a method of cultivating callus of a plant belonging to the genus Crocus on an MS medium supplemented with plant hormones. The use of plant hormones such as auxin and cytokinin for culturing plant tissues has been well known for a long time, and Murashige-Skoog medium is also a commonly used medium.

[0007] JP-A-63-258574 discloses that the stigma, style, ovary, ovule, and petals of saffron are excised and cut and placed in a LS medium or a B5 medium supplemented with cytokinin and auxin. It is transplanted and subcultured. Extraction, cutting and cultivation of plant parts have been performed for a long time, and plant hormones such as cytokinin and auxin are usually used, and Linsmeier Skoog's medium and Gamborg's medium are also commonly used media.

Tests of any of these methods do not significantly increase the rate of growth of the active ingredient of saffron and do not significantly increase the production of the active ingredient.

[0009]

SUMMARY OF THE INVENTION An object of the present invention is to provide a method for more efficiently obtaining pigments of crosine and crocetin, which are one of the active ingredients of saffron.

[0010]

Means for Solving the Problems As a result of intensive studies to solve the above-mentioned problems, the present inventors have found that it can be solved by adding methyl salicylate to the medium, and have completed the present invention. That is, the present invention is a method for cultivating a tissue of saffron veins comprising culturing using a medium containing methyl salicylate.

The amount of methyl salicylate is 1 to 10000 ppm, preferably 10 to 1000 pp, based on the culture medium.
It has been found that the addition of m gives good results.

The medium used for this purpose is not particularly limited, but may be Murashige and Skoog medium, White medium, Gamborg et al. Medium, Nitsch medium, Heller medium. Medium, Schenk and Hildebra
ndt), Nitsch and Nitsc
h) Medium, Kohlenbach and Schmidt (Kohllenbac
k and Schmidt's medium, Knop's medium, and Linsmaier and Skoog's medium.

If necessary, cytokinins and auxins known as plant growth regulators may be added to these media. Examples of cytokinins include benzyladenine, zeatin, kinetin, and the like, which vary depending on the type.
Add 0 ppm. Examples of the auxin include naphthalene acetic acid, indole acetic acid, 2,4-dichlorophenoxy acetic acid and the like. The auxin varies depending on the type thereof, but when added, 0.01 to 20 ppm is added.

[0014] If necessary, other medium-forming agents can also be added. The culture is performed at a temperature of 20 to 25 ° C. in a dark place. The inoculated mesibet usually turns brown in 2 to 4 weeks, but by culturing under these conditions, it does not turn brown even after 6 months, grows normally, and the amount of pigment increases, It turned out that the red part was more red, and the white part was also red.

[0015]

EXAMPLES The present invention will be described more specifically with reference to the following examples, but the present invention is not limited to these examples. (Examples 1 to 5) (Comparative Examples 1 to 16) The outer skin of a saffron bulb with flower buds extending to about 10 cm was peeled off, the discolored portion was removed, and the saffron was washed with running water for 3 hours. 3-4 seconds for 10% benzalkonium chloride, 1-2 seconds for 70% ethanol, 5
% Sodium hypochlorite for 20 to 40 minutes. After that, it was performed aseptically. The flower buds were cut off leaving the bulbs, and washed three times with sterile water. Next, the buds were taken out from the flower buds, incised, and the mesibes were taken out with the style long, and inoculated on the medium. The medium was Murashige and Skoog's medium, 4% sucrose, 0.8% agar powder, pH 5.8.
Was adjusted. To this was added 10 ppm of naphthalene acetic acid and 1 ppm of benzyl adenine. The additives shown in Table 1 were further added thereto. Table 2 shows the culture results.

[0016]

[Table 1]

[0017]

[Table 2]

(Examples 6 to 10) (Comparative Examples 17 to 32) In the medium of Examples 1 to 5, naphthalene acetic acid was changed to 7 ppm, and benzyl adenine was changed to 0.5 ppm. Cultures were performed with additives. Table 4 shows the culture results.
Shown in

[0019]

[Table 3]

[0020]

[Table 4]

[0021]

According to the present invention, the addition of methyl salicylate to the culture medium of saffron mesibes increases the amount of pigment and suppresses browning over a long period of time, as shown in Examples.

──────────────────────────────────────────────────の Continued on the front page (51) Int.Cl. ⁷ Identification code FI (C12P 19/44 C12R 1:91)

Claims (1)

(57) [Claims] 1. A method for cultivating a tissue of Saffron Messibe, comprising culturing in a medium containing methyl salicylate.