JPS6339517A - Plant tissue culture method and container - Google Patents

Abstract

(57) [Abstract] This bulletin contains application data before electronic filing, so abstract data is not recorded.

Description

[Detailed Description of the Invention] [Industrial Application Field] This invention aims to continuously produce large quantities of plants with the same properties and uniform grains as their parents by extracting and culturing only excellent cells and Mi fibers. The present invention relates to a plant tissue culture method and a culture container thereof.

[Conventional technology]

In recent years, when breeding and improving plant varieties, unlike traditional plant breeding methods such as cuttings and division, the plant tissue culture method has been used as a technology for mass production by passing on the excellent traits of YT to the next generation. is in the spotlight.

The advantage of this method is that it can be cultured and grown from plant cells on a laboratory scale.

The current plant tissue culture method uses a container with an opening at the top, such as a hard Erlenmeyer flask, a culture flask, or a large cylindrical hard test tube, as a culture container, and places the plant tissue in it. The upper opening is closed with a lid or stopper such as silicone resin foam, aluminum foil, a rubber plug filled with perforated cotton, a silicone resin cap, or a stainless steel cap, thereby preventing the intrusion of germs and allowing ventilation. It is cultivated with the aim of

[Problem that the invention seeks to solve]

However, as described above, all of these culture containers have an opening at the top of the container.

The opening is extremely narrow, and the opening is closed with a silicone stopper or aluminum foil that has a ventilation hole.

Therefore, it is extremely difficult to implant cultured plants into these culture containers, to transfer the plants, etc., and because the opening of the culture container is located upward, and the opening is closed with the above-mentioned lid or stopper, This has the disadvantage of blocking light from above, causing poor growth and fading.

Furthermore, when these culture containers are used, they are sterilized by heating the entire container, but this heating causes water droplets to adhere to the inside of the container after sterilization.

Therefore, it is necessary to completely remove the water droplets attached to the inside of the culture container, but since the opening of the culture container is extremely narrow, it is not possible to completely remove the water droplets, which reduces the amount of light. It is also a cause.

Furthermore, since insufficient light to the culture container is likely to occur, 2.
To compensate for the lack of light due to these various factors, it is necessary to take measures such as increasing the number of light sources on the culture container or moving the culture container closer to the light source. The cultivation container is exposed to higher temperatures than necessary, causing the troublesome problem of high temperature damage to the plants inside the container.
Another disadvantage is that cultivation costs are high.

[Means for solving problems]

In view of the current situation, this invention has been developed to
The purpose is to facilitate work such as planting plants by using a new culture container in which the M body and the container body can be separated, and to perform plant tissue culture under excellent lighting conditions. It is.

The first invention consists of a hollow transparent lid body having a culture space inside, and a wide-mouthed container body formed with a locking groove capable of locking an opening of the lid body on the outer periphery of a culture medium storage part. This is a plant tissue culture method characterized in that plant tissue culture is performed using a culture container constructed as described above.

Further, the second invention includes a hollow transparent lid having a culture space therein, and a wide-mouthed lid having a locking groove for locking the opening of the lid on the outer periphery of the culture medium storage section. It consists of a container main body, and an air intake port is formed in a part of the container main body, and the culture is formed between the lid body and the container main body only through a filter attached to the air intake port. This plant tissue culture container is characterized in that it is configured to allow ventilation without the intrusion of germs into the space.

That is, the plant tissue culture method in the first invention more specifically includes a lid body that can be divided into upper and lower parts;
Plant tissue culture is carried out using a culture container consisting of a container body.A solid medium is stored in the medium storage section of the container body, and after inoculating cell masses (callus) etc. into this solid medium, .

Only the 9M body has a larger culture space in order to enlarge the culture space in the culture container according to the growth degree of callus, etc., with a hollow transparent lid having the above-mentioned culture space attached to the container body. This is a method of culturing and growing by replacing the lid.

Furthermore, the second invention relates to the structure of the plant tissue culture container for use in the first invention, and the lid body constituting the culture container that can be divided into one part is formed of a transparent material such as plastic or glass, or a colored transparent material, The interior thereof has a size and/or shape that can substantially constitute the culture space,
Moreover, this transparent body has a structure in which there is no part that obstructs light from entering the culture space.

One container body is provided with a wide-mouth culture medium storage part having a storage part for a solid medium such as an agar medium, and a locking groove that can lock the opening of the lid body formed on the outer periphery thereof, The container body has an air intake port for supplying outside air to the culture space, and its material is, for example, polypropylene or polyethylene.

Tetrafluoroethylene resin, silicone resin, glass.

Alternatively, it is made of a material such as ceramic.

Further, a filter is attached to the air intake port provided in the container body, and this filter is made of a material such as silicone resin foam, cotton paper, porous ceramic, or glass wool.

[For production]

The plant tissue culture container 1 of the present invention includes a hollow transparent lid 2 having a culture space 5 therein, and a lock for locking the opening of the lid 2 to the outer periphery of the culture medium storage section 8. It is composed of a wide-mouthed container body 7 in which a groove 10 is formed.

Therefore, when culturing plant tissue, a solid medium 13 such as an agar medium is stored in the medium storage part 8, and after plant cells or callus are planted in this solid medium 13, the medium storage part 8 is covered and the container is placed. The hollow transparent lid body 2 is locked to the main body 7 in a substantially hermetically sealed state, and only the air taken in through the air intake port 11 consisting of a vent hole provided in the container main body 7 is purified by the filter 12 to form a culture container. At the same time, the cells or callus, etc., are cultured and grown in a dark place, etc., while irradiating them with light for a predetermined period of time if they require light, or in the dark if they do not require light. be.

Especially when culturing and growing by irradiating light,
The irradiated light is efficiently absorbed into the culture space 5 from the entire surface of the transparent lid 2.

Furthermore, prior to tissue culture, the culture container 1 is sterilized either alone or with the solid medium 13 housed therein.
In particular, when sterilization is performed with the solid medium 13 stored, even if water droplets adhere to the inside of the lid 2 due to moisture evaporating from the solid medium 13, these water droplets can be easily removed. I can do it.

There will be no shortage of light due to water droplets or the like.

When culturing and growing plant seeds, callus, etc. in the culture container 1, the generated seedlings are exposed to sufficient light and moderate temperature from the entire surface of the hollow transparent lid 2 into the culture space 5 of the culture container 1. Clean air containing no germs is supplied to the culture space 5 only through the filter 12 from the air intake port 11 formed in the container body 7, such as a vent hole.

The culture space 5 is always kept under the optimum conditions for growth, and cells etc. can differentiate into stems and leaves, extend filamentous roots, and achieve ideal growth.

Furthermore, in this plant tissue culture method, the culture container 1
By appropriately replacing only the hollow transparent lid 2 constituting the container with a large hollow lid that can gradually enlarge the culture space 5 as the plants grow, plant growth is not hindered. It is also possible to maintain this condition and promote growth.

〔Example〕

Hereinafter, one embodiment of the plant tissue culture container of the present invention will be described in detail based on the accompanying drawings.

A method for culturing plant tissue using the above plant culture container will be explained.

FIG. 1 is a longitudinal cross-sectional view showing a culture container of the present invention and an example of culturing plant tissue using this culture container.

This plant tissue culture container 1 has a lid 2 that can be divided into 9 parts.
and the container body7. and a filter 12.

The lid body 2 is made entirely of a transparent body made of glass or plastic, and consists of a spherical hollow part 3 with a culture space 5 formed therein, and a cylindrical skirt part 4 formed at the bottom of the hollow part 3. An opening 6 is formed by the portion 4.

The container body 7 is made of 2 materials such as polypropylene, polyethylene,
Tetrafluoroethylene resin, silicone resin.

It is made of a material such as glass or ceramic, and is located on the outer periphery of a flat cylindrical culture medium storage section 8 with a relatively wide mouth as shown in FIG.

A cylindrical body 9 having a peripheral wall 9a lower than the peripheral wall 8a of the culture medium storage part 8 is provided on a concentric circle, and the peripheral wall 8 of the culture medium storage part 8 is provided on a concentric circle.
A ring-shaped locking groove 10 for loosely fitting and locking the skirt portion 4 of the lid body 2 is formed between the peripheral wall 9a of the outer cylinder 9 and the circumferential wall 9a of the outer cylinder 9. Peripheral wall 9a in places
An air intake port 11 consisting of a ventilation hole is provided through the culture space 5, and only outside air from the air intake port 11 is supplied to the culture space 5 via a filter 12, which will be described later.

Therefore, in the case of this embodiment, the culture medium is made such that a predetermined space is always maintained between the peripheral wall 8a of the culture medium storage section 8 and the outer peripheral surface of the cylindrical skirt portion 4 of the lid body 2. It is necessary to make the inner diameter of the skirt portion 4 larger than the outer diameter of the storage portion 8.

The filter 12 installed in the engagement groove 10 of the container body 7 is made of a porous material such as silicone resin foam, glass wool, cotton paper, or porous ceramic, and is shown in FIGS. 1 and 3. Engagement groove l as shown in
A cross-sectional cylindrical body having a cylindrical body integrally provided on the outer peripheral edge of a ring member having the same width as the width of the air intake port 11 and having a thickness that covers the opening surface of the air intake port 11. It is the body.

In order to perform tissue culture of plants using a culture container having the above structure, first, as shown in FIG. For example, conventionally known agar,
The main material is a basic medium containing S. cerevisiae, etc., and various drugs such as vitamin B, amino acids, plant growth hormones, etc.
After storing the solid medium 13 containing nutrients and obtained by a conventional method, a flat lid 2a that hardly forms the culture space 5 is placed above the medium storage part 8 to sterilize the medium. The lid body 2 having the large hollow part 3 forming the culture space 5 above the storage part 8 is separately sterilized.

In this case, if it is desired to culture and grow using the lid 2 with a large hollow portion 3 from the beginning, the container body 7 and the lid 2 may be sterilized at the same time.

After that, a plant tissue 2 that has been sterilized in advance, such as sterilized plant cells or callus, is placed on the solid medium 13, and the skirt portion 4 of the 9M body 2 is locked in the locking groove 10 of the container body 7, and the growth begins. The air necessary for and a filter 12 from an air intake port formed by a gap a formed by the circumferential wall 9a of the cylinder body 9 and the skirt portion 4 of the lid body 2.
The cells are supplied to the culture space 5 through the space formed between the peripheral wall 8a of the culture medium storage part 8 and the inner wall of the skirt part 4 in a cleaned state, and the cells and the like are grown in the culture container 1. It is something to do.

Note that when plant cells or callus are initially planted in the solid medium 13, they do not require much air, so the fourth
1 to 3 using the flat lid body 2a as shown in the figure.
The cells are cultured and grown for about a week, and then depending on the degree of growth of the cells, etc., the lid body 2 of the culture space 5 is large. That is, culture and growth may be performed by replacing the lid body 2 with a large hollow part 3.

At that time, if the container body 7 is fitted and mounted on a highly stable base 14 as shown in FIG. 1, the culture container 1 can be kept stable even if the culture space 5 of the lid 2 used is large. It can be used in the same condition.

If the plants to be cultured are ornamental flowers such as carnations, gypsophila, orchids, the culture container 1 can also be used as a plant identification container by considering the design of the lid 2 in advance. It is something that can be done.

Even if the lid 2 constituting the culture container 1 is entirely spherical as in the above embodiment, it may have a mushroom-shaped hollow part 3 as shown in FIG. 5, or as shown in FIG. It may be a type having a right cylindrical hollow part 3 as shown in the figure, and the key point is to use it according to the shape of the plant to be cultured, the size at maximum growth, the growth stage, the work process during cultivation, etc. in.

If necessary, several types of lids with different sizes of hollow portions 3 may be prepared in advance and used by replacing them according to the conditions described above.

Further, the shape of the container main body 7 is not limited to the above-mentioned embodiment. For example, as shown in FIGS. 8b
, 8b facing each other in the vertical direction, and having a peripheral wall 9a of the same height on the outside of the peripheral wall 8a. are placed on concentric circles.

A locking groove 10 for locking the skirt portion 4 of the lid body 2 is formed between this peripheral wall 9a and the peripheral wall 8a of the medium storage portion 8 to constitute the container body 7. An air intake port 11a consisting of a vent hole communicating with the vent hole 8b provided in the culture medium storage section 8 may be provided at the bottom, and a filter 12 may be installed in the air intake port 11a. Well, the point is to send air into the culture space 5 formed by the culture medium storage part 8 and the lid body 2 through the container body 7, which has a locking groove 10 for locking the lid body 2 on the outside of the medium storage part 8. There is no particular restriction on the structure as long as it has an air intake port 11 that allows air to pass through the culture space 5 only through the filter 12 and allows ventilation to flow into the culture space 5 without invading germs.

Furthermore, the shape of the filter 12 is also as shown in FIG.
A cylindrical body is integrally provided on the inner peripheral edge of a ring member that has the same width as the groove width of the engagement groove 10 and has a thickness that covers the opening surface of the air intake port 11. As a type of cylindrical body,
After this filter 12 is installed in the locking groove 10 of the container body 7, the outer circumferential wall of the filter 12 and the inner circumferential wall 9 of the cylinder body 9 are
The lid 2 may be fixed to the container body 7 by locking the skirt portion 4 of the lid 2 in the groove formed between the grooves a.

〔Effect of the invention〕

The plant tissue culture method of the present invention includes a hollow transparent lid body having a culture space inside, and a wide mouth container body having a locking groove for locking the lid body on the outer periphery of a culture medium storage part. After a plant tissue is placed in a culture medium stored in a culture medium storage part of the culture container, the lid is locked to the container body to perform tissue culture of the plant.

Therefore, if the mouth of the container, such as a hard Erlenmeyer flask or a large hard test tube, is extremely narrow and the distance between the mouth and the culture medium is relatively long, conventional methods that use a cap or stopper at the mouth of the container may be difficult. Compared to the culture method, the culture container itself can be divided as necessary, making it extremely easy to place cells, etc. on the bed, which greatly improves work efficiency. If necessary, light can be absorbed from the entire surface of the lid.

Therefore, the amount of light required for plant tissue culture can be sufficiently secured with a small light source, and the cost for culturing can be significantly reduced, and there is no risk of inducing poor growth or fading.

Also according to the plant growth stage and working process.

By replacing only the -41i body that is locked to the container body with a lid body of a different size and shape, the cultivation can be made more reliable and the work can be carried out more easily.

Furthermore, the plant tissue culture container of the present invention is composed of a container body having a locking groove on the outer periphery, and a hollow transparent lid that covers the culture medium storage part of the container body to form a culture space, In addition, the container body has an air intake hole that allows ventilation into the culture space without bacteria entering through the filter, so that by using this culture container, the plant tissue culture method having the above-mentioned characteristics can be achieved. can be carried out.

In addition, this culture container has a hollow, transparent lid that provides sufficient light and an appropriate temperature within the culture space, and at the same time, bacteria are removed only through a filter from the air intake hole formed in the container body. Since clean air can be supplied to the culture space, the culture space can be placed in the optimal environment for growth.

In addition, this plant tissue culture container has a divided structure consisting of the container body and lid, which makes it possible to prevent insufficient light intensity due to adhesion of water droplets etc. to the inner surface of the 9M body, and also to reduce maintenance work You can make it happen.

Furthermore, it is also possible to replace the fluid with one of a different size and/or shape as the plant grows, etc.
These can maintain conditions that do not hinder plant growth.

Note 2: This plant tissue culture container has a hollow, transparent lid with various designs, so that it is possible to store a single tissue-cultured plant in the culture container and sell it for plant appreciation. It has a great effect on tissue culture.

[Brief explanation of the drawing]

FIG. 1 is a longitudinal cross-sectional view showing an example of the structure of the culture container of the present invention, and illustrating the cultivation of plants Mi and textiles using this culture container. FIG. 2 is a perspective view of the container body in FIG. 1, and FIG.
The figure is a partially enlarged longitudinal cross-sectional view showing the locked state of the lid and the container body. Figure 4 shows the combination of the container body and the flat lid.
FIG. 5 is a vertical cross-sectional view showing i-woven culture, FIG. 5 is a vertical cross-sectional view showing another shape of the fluid, and FIG. 6 is a vertical cross-sectional view showing another shape of the lid body. FIG. 7 is a partially enlarged vertical cross-sectional view showing another example of the structure in which the lid and the container body are engaged, and FIG. 8 is a partially cutaway plan view showing another example of the structure of the container main body. FIG. 9 is a longitudinal sectional view of FIG. 8. 1... Plant tissue culture container 2... Lid body 3... Hollow part 4... Skirt part 5... Culture space 6... Opening part 7... Container body 8... Culture medium storage Part 9...
Cylindrical body 10...Latching groove 11...Air intake port 12...
・Filter 13...Solid medium 14...Base

Claims (6)

[Claims] (1) A hollow transparent lid body with a culture space inside;
The present invention is characterized in that plant tissue culture is carried out using a culture container consisting of a wide-mouthed container body in which a locking groove capable of locking the opening of the lid body is formed on the outer periphery of the culture medium storage section. Plant tissue culture methods. (2) In the above plant tissue culture method, tissue culture is performed by replacing only the lid of the culture container used so that the size and/or shape of the internal culture space can be varied depending on the growth stage of the cultured plant and the work process. The plant tissue culture method according to claim 1, characterized in that: (3) a hollow transparent lid body with a culture space inside;
It consists of a wide mouth container body in which a locking groove capable of locking the opening of the lid body is formed on the outer periphery of the culture medium storage part, and an air intake port is formed in a part of the container body. Plant tissue culture, characterized in that the culture space formed between the lid body and the container body can be ventilated only through a filter attached to the air intake port without intrusion of germs. container. (4) The container body is made of a material selected from polypropylene, polyethylene, tetrafluoroethylene resin, silicone resin, glass, and ceramic. The described plant tissue culture vessel. (5) The plant tissue culture container according to claim 3, wherein the lid is made of a transparent material such as glass or plastic, or a colored transparent material. (6) The filter used in the culture container is made of a material selected from silicone resin foam, cotton, paper, glass wool, and porous ceramic. The plant tissue culture container described in Section.