JPH0638A - Method for culturing seedling of orchis graminifolia - Google Patents

Abstract

PURPOSE:To obtain a method for efficient culture suitable for large amount of culture in a short culture period. CONSTITUTION:Protocorms or protocorm-like materials of Orchis graminifolia are fed to a culture tank charged with a liquid medium in a culture device and sterile air is passed through the liquid medium. Protocorms or protocorm- like materials of Orchis graminifolia are cultured while irradiating with light rays to raise seedlings.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION The present invention relates to a method for raising a moth orchid seedling by culturing a moth orchid protocomb or a protocomb-like body.

[0002]

2. Description of the Related Art Phalaenopsis orchid seedlings are generally obtained by sowing seeds that have been crossed on agar medium containing various nutrients in a flask, sprouting and rooting through a protocomb, and then in the flask until they reach a certain size. Raise seedlings while maintaining aseptic conditions. During this period, several times of aseptic transplantation work are required to prevent overcrowding of seedlings and removal of poorly grown seedlings, and the culturing period is extremely long, 6 to 10 months.

Recently, the production of cloned seedlings of Phalaenopsis orchid is also increasing, and in this case, leaf pieces or flower stem axillary buds are cultured to form protocome-like bodies, which are proliferated to increase the number of Similar to the method, the seedlings are raised by placing them on an agar medium containing various nutrients in a flask, allowing them to sprout and root. Even in this case, transplantation work and a long culture period are required.

[0004]

According to the conventional method,
The work of transplanting from flask to flask must be done manually by each strain, which is extremely time-consuming.
Also, since the culture period is long, a large culture room was required. Furthermore, there was almost no merit of scale-up by carrying out mass culture, resulting in a costly seedling culture method, which was not suitable for mass culture.

Further, it takes a long time to raise seedlings, which means confirmation of mutations during production of cloned seedlings and prolongation of breeding cycle, and shortening of culture period is strongly desired. An object of the method of the present invention is to reduce the number of cultivation steps and culture cost in cultivating these phalaenopsis seedlings and to cultivate an efficient phalaenopsis seedling with a short culture period.

[0006]

Means for Solving the Problems As a result of intensive studies in view of such circumstances, the inventors of the present invention have introduced a protocome or protocomb-like body of Phalaenopsis orchid into a liquid medium of a culture device and sterilized it in a liquid medium. By aerating the above air and culturing while culturing while irradiating with light, the absorption efficiency of nutrients is improved, growth is effectively promoted by sufficient oxygen supply, and further the labor of transplantation is reduced. It was found that the three-dimensional mass culture was possible and the method of the present invention was completed.

The culture device used in the method of the present invention comprises a culture tank containing a liquid medium, a sterile air supply device, a temperature control device, and a lighting device. When culturing in a temperature-controlled room, the temperature adjusting device can be omitted. In addition, since the stirring blade damages the plant body, it is desirable to use only stirring by aeration. The light source of the lighting device may be an artificial light such as a fluorescent lamp or a high-pressure sodium lamp, and may be outside or inside the culture tank. Also, the culture is performed at a constant temperature of 20 to 35 ° C. and 100
It is preferable to carry out under irradiation with light of 0 to 5000 lux.

The liquid medium used in the method of the present invention is used as a carbon source in a medium containing various nutrients necessary for plant growth, such as Murashige-Skoog medium, Hyponex medium, Camborg medium, or a modified medium thereof. It is possible to add sugars such as sucrose and glucose in an amount of 5 to 30 g / l. Although it is not necessary to add plant hormones such as auxin and cytokinin, 5 to 30% by weight of natural juices such as coconut water, apple, banana, tomato and potato may be added.

The moth orchid plant tissue used in the method of the present invention may be a protocomb formed from seeds or a protocomb-like body formed from leaf pieces, flower stem axillary buds and the like.

[0010]

According to the method for cultivating a phalaenopsis orchid according to the method of the present invention, the protocome or protocomb-like body of the phalaenopsis orchid is efficiently absorbed with nutrients in a liquid medium containing sufficient dissolved oxygen, so that it is extremely grown. It is fast and can significantly shorten the culture period. In addition, since it is cultivated in a liquid medium, the transplanting operation is easy, and three-dimensional culturing is possible, so it is possible to produce a large amount of seedlings at one time, and by performing scale-up, the number of culturing steps and The cost can be reduced, and the culture method is suitable for large-scale culture.

[0011]

【Example】

(Example 1) 6 parts of Hyponex fertilizer (N.P.K. = 6.5.6.19) were placed in a 1-liter glass aeration type culture tank.
900 ml of a medium containing g / l and 15 g / l of sucrose was added and adjusted to pH 5.3, and then sterilized by autoclaving at 120 ° C. for 20 minutes.
10 protocorm-like bodies formed from the axillary buds of the flower stalks of the white-flowered yellow-lip moth orchid were placed in the chamber, and irradiated with 2000 lux of daytime fluorescent light for 14 hours in a temperature-controlled room at 26 ° C. Culture was performed for a month. As a result, the average leaf elongation was 4.14 cm / strain growth.

(Example 2) In a 0.9-liter aeration culture tank made of glass, Hyponex fertilizer (NPK = 6.
5 ・ 6 ・ 19) 3g / l, sucrose 15g
Pour 600 ml of the culture medium containing the
After adjusting to H5.3, autoclave sterilization was performed at 120 ° C. for 20 minutes. 10 protocomes formed from white-flowered yellow lip moth orchid seeds were placed in the room, irradiated with 2000 lux of 14-day photoperiod fluorescent light in a temperature-controlled room at 26 ° C., and sterilized air was aerated to incubate for 2 months. went. As a result, the average leaf elongation was 5.85 cm / strain growth.

(Comparative Example 1) In a 300 ml Erlenmeyer flask, 60 ml of 0.8% agar added to the medium having the same composition as in Example 1 was added, and after adjusting the pH to 5.3 in the same manner, the autoclave was sterilized. The agar solid medium was prepared. On this, 10 protocomb-like bodies formed and proliferated from axillary buds of phalaenopsis of white flower yellow lip were placed,
The cells were cultured for 2 months by irradiating them with a fluorescent light of 2000 lux having a photoperiod of 14 hours in a constant temperature room at 26 ° C. In addition, during this period, one month was subcultured once in a completely similar medium. As a result, the average leaf elongation was 1.48 cm / strain growth.

(Comparative Example 2) A 300 ml Erlenmeyer flask was charged with 60 ml of a medium having the same composition as in Example 2, to which 0.8% of agar was added, and after adjusting the pH to 5.3 in the same manner, the autoclave was sterilized. The agar solid medium was prepared. 10 pieces of protocomes formed from white orchid lip lip moth orchid seeds were placed on this, and they were irradiated with 2000 lux of fluorescent light with a photoperiod of 14 hours in a constant temperature room at 26 ° C.
Culture was carried out for 2 months. In addition, during this period, one month was subcultured once in a completely similar medium. As a result, the average leaf extension amount was 3.21 cm / strain growth.

[0015]

According to the method of the present invention, the moth orchid protocomb or protocomb-like body is put into the liquid medium of the culture apparatus, and the liquid medium is cultivated while aerating with sterile air and irradiating with light. By raising seedlings, the absorption efficiency of nutrients is improved, and sufficient oxygen supply can be expected to promote the growth of phalaenopsis seedlings. In addition, since the culture is performed in a liquid medium, the labor of transplantation and three-dimensional mass culture can be achieved.

Claims (1)

[Claims] 1. A moth orchid protocomb or a protocomb-like body is put into a liquid medium of a culture device, sterile air is aerated to the liquid medium, and the seedlings are grown by culturing while irradiating with light. Method for cultivating Phalaenopsis seedlings.