JP2517322B2 - Growth promoter for orchids - Google Patents

Description

TECHNICAL FIELD The present invention relates to a growth promoter for orchids.

Conventional technology and its problems Conventionally, in tissue culture of orchids, in order to promote rooting and growth of orchids, in a tissue culture medium, as a growth promoter, an auxin plant hormone, A cytokinin plant hormone and coconut oil (coconut milk) are mixed and added. Although these growth promoters show excellent effects when used in combination with each other, they have drawbacks that production, reaction, separation and purification are difficult and cost is high because they are chemically synthesized products. . Moreover, in order to sufficiently bring out the effect, it is required to add a very small amount to each, so that handling operations such as weighing become complicated.

Means for Solving the Problems The inventors of the present invention have conducted extensive studies in view of the above-mentioned problems of the prior art, and as a result, (a) a hot water extract of Euglena cells, which are protozoa, is used for orchids. Conventional growth promoter 3
It shows an even better growth-promoting effect than in the case of using seeds in combination. (B) Even if the addition amount is selected from a reasonably wide range where handling is not complicated, the effect will not change at all. Heading, completed the present invention.

That is, the present invention relates to a growth promoter for western orchids, which comprises a hot water extract of Euglena cells as an active ingredient.

The protozoan euglena used in the present invention retains chloroplasts in its cells, normally lives in fresh water such as rivers and lakes, and is used as an autotrophic and external environment having a photosynthetic function using carbon dioxide as a carbon source. It has a life stage of heterotrophic with existing organic substances as carbon sources. Examples of euglena include euglena gracilis and euglena
Examples include all Euglena species such as virgin, Euglena intermidiae and other Euglenoids, wild strains inhabiting rivers and lakes, and mutants thereof. In addition, there is no restriction on the medium and culture form, and any of those that are heterotrophically cultured in the dark and those that are cultured under photosynthetic conditions under irradiation of sunlight or artificial light can be used. is there. Of course, by changing the medium and culture conditions,
Although the components in the Euglena cells change somewhat, there is no obstacle to achieving the object of the present invention.

The hot water extract of Euglena cells cultured by the above method can be produced, for example, as follows.

First, the Euglena cells cultured by the above method are recovered from the culture broth according to a known microbial cell recovery method such as centrifugation or overseparation. Then, the recovered Euglena cells are washed with tap water, pure water, ion-exchanged water or the like a suitable number of times (usually about 2 to 3 times), if necessary, and then a known method such as centrifugation or gravity sedimentation method. The water and the Euglena cells are separated with and the obtained Euglena cells are dried.
A known method can be adopted as a drying method, and examples thereof include freeze gliding, hot air drying, and natural air drying. The dried product of Euglena cells (hereinafter referred to as dried cells) thus obtained is subjected to the following hot water treatment. The recovered Euglena cells and the Euglena cells after washing with water (hereinafter referred to as wet cells) can also be used as a raw material for hot water treatment.

The hot water treatment is carried out by heating an aqueous suspension of dry cells or wet cells under pressure, if necessary.
The amount of dry cells or wet cells in the water suspension is not particularly limited, but usually the weight ratio (cell / water) may be about 1/20 to 1/2. The heating temperature and time are not particularly limited, but the heating temperature is usually about 60 to 120 ° C., and the heating time is about 5 to 120 minutes. In the present invention, 60 to 120 ° C
Hot water is water at about the same temperature. After cooling the aqueous suspension of dry cells or wet cells treated with hot water to room temperature, the solid content in the aqueous suspension is removed by a known separation method such as centrifugation or excess to remove heat of Euglena. A water extract solution is obtained.

In the present invention, the hot water extract solution of Euglena obtained above may be added to the medium of Western orchids as it is or after diluting or concentrating it, or may be added as a dried product according to a known method. Good, but when the ratio of cells to water used in hot water extraction is the above ratio (1/20 to 1/2), dilute the hot water extract obtained with water to about 10 to 10,000 cultures. Is preferably used.

The tissue culture of the orchids using the Euglena hot water extract of the present invention may be carried out according to a known method, for example, as follows.

As a tissue culture medium for western orchids, any of those conventionally used can be used, for example, Murashige-Skoog medium (hereinafter referred to as MS medium),
Examples thereof include a white medium and a medium obtained by adding various commercially available inorganic fertilizers to fruit juice such as apple juice.

In the present invention, a hot water extract of Euglena cells is added to the above medium to obtain the tissue culture medium for western orchids of the present invention. The addition amount of the hot water extract of Euglena cells is not particularly limited, but Euglena cells and water are added in the above ratio (1/20 to 1/1).
When a hot water extract obtained by the hot water extraction treatment in (2) is used, it is sufficient that the extract is usually contained in the medium in an amount of about 10 to 100000 ppm. If it is less than 10 ppm, the growth promoting effect of orchids tends to be insufficient, while
If it exceeds 100000ppm, the growth of orchids tends to decrease.

The thus obtained culture medium for tissue culture of western orchids may be filled and sterilized in the same manner as in the conventional method, and then inoculated with a suitable piece of tissue of western orchids and cultured.

EFFECTS OF THE INVENTION The hot water extract of Euglena cells of the present invention exhibits a growth promoting effect on orchids which is superior to that of conventional growth promoters. In the case of culturing western orchids using the extract, the effect will be exhibited even if the addition amount is selected from a reasonably wide range so that handling is not complicated.

Examples The following examples and comparative examples will further clarify the present invention.

Example 1 1) Preparation of hot water extract of Euglena cells 50 g of a dry product of Euglena was suspended in 300 ml of tap water and subjected to hot water extraction at a temperature of 120 ° C. for 10 minutes in an autoclave, followed by centrifugation (10000 × G). ) To remove solids to obtain an extract of Euglena cells.

2) Preparation of tissue culture medium Solution A NH ₄ NO ₃ 26.4g KNO ₃ 30.4g CaCl ₂・ 2H ₂ O 7.0g MgSO ₄・ 7H ₂ O 5.9g KH ₂ PO ₄ 2.7g And was finally set at 4.

Liquid B MnSO ₄・ 4H ₂ O 25.0g H ₃ BO ₃ 10.0g ZnSO ₄・ 7H ₂ O 10.0g KI 1.0g Na ₂ MoO ₄・ 2H ₂ O 250.0mg CuSO ₄・ 5H ₂ O 25.0mg CoCl ₂・ 6H ₂ O 25.0 mg The above components were dissolved in distilled water 1.

Solution C Myo-inositol 10.0 g Nicotinic acid 500.0 mg Glycine 200.0 mg Vitamin B ₁ 50.0 mg Vitamin B ₆ 50.0 mg d-Biotin 5.0 mg The above components were dissolved in distilled water 1.

D liquid 50.0 mg of folic acid was dissolved in 100 ml of distilled water.

Solution E Na ₂ · EDTA 7.46 g FeSO ₄ · 7H ₂ O 5.56 g The above components were dissolved in distilled water 1.

A solution 250 ml, B solution 1 ml, C solution 5 ml, D solution 1 ml, E solution 5 ml,
Stock solution of Euglena cells extracted with sucrose 10g, agar 8g and 1) hot water extraction stock 100ml (dry weight 21.7g), 10ml or 1ml
Water was added to the whole to prepare a tissue culture medium for 1 year (pH 5.4).

3) Tissue culture The medium obtained in 2) was placed in 20 glass flat-bottom test tubes each having a diameter of 30 cm and a length of 150 cm, 20 ml each, and sterilized by autoclaving at 120 ° C for 15 minutes. After cooling, unrooted seedlings were planted in each test tube. As the unrooted seedlings, those obtained by cultivating the meliclon of the apex of the stem of a Western orchid of the genus Dendrobium were used. After planting, 90 at a temperature of 20-23 ℃
After culturing for a day, the rooting state (root length and number of roots) was examined. The results showed that the root length was 1.
The number was 48 to 1.52 cm and the number of roots was 7.1 to 8.0 (standard deviation 0.701).

Example 2 Instead of the hot water extraction stock solution, dried products of the hot water extraction stock solution 10 g, 1 g
Alternatively, the culture was performed in the same manner as in Example 1 except that 0.1 g was used. The results showed that the root length was 1.43 to 1.46 cm and the number of roots was 7.5 to 8.2 (standard deviation 0.750), almost independent of the amount of dry matter added.

Comparative Example 1 Culture was performed in the same manner as in Example 1 except that the hot water extraction stock solution was not used. The root length was 0.674 cm, and the number of roots was 5.7 (standard deviation 0.563).

Comparative Example 2 Instead of the hot water extraction stock solution, plant hormones (auxin-based and cytokinin-based) 1 ppm each and coconut milk 15
Cultivation was carried out in the same manner as in Example 1 except that 100% was used. The root length was 1.420 cm and the number of roots was 7.5 (standard deviation 0.808).

Claims (1)

(57) [Claims] 1. A growth promoter for western orchids, which comprises a hot water extract of Euglena cells as an active ingredient.