JPH07184497A - Germ-free capsule for ornamental plant - Google Patents

Abstract

PURPOSE:To provide germ-free capsules for ornamental plants being fit for admiration for a prolonged period of time with no infiltration of microorganisms and no fogging with moisture. CONSTITUTION:Ornamental plants 6 such as tissue-cultured TSUWABUKI (Japanese silver leaf) or coffee plant aseptically grown from seeds are transplanted to the culture medium 5 in a capsule made of a clear, heat-resistant polycarbonate resin. At the top of the capsule cover 2, an air vent hole 3 is bored in a diameter of 2 to 6mmphi. The vent hole 3 is blocked with an air vent film 4 which inhibits the infiltration of microorganism floating in air, but permits oxygen and carbon dioxide gases concerning to the plant metabolism to pass through freely. The air vent film 4 is, for example, a porous membrane film of polytetrafluoroethylene of 10 to 5mum maximum pore size and 60 to 80% porosity.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to an aseptic plant aseptic capsule, and more particularly to an aseptic plant aseptic capsule capable of enduring long-term appreciation.

[0002]

2. Description of the Related Art An ornamental plant container for observing a sterile plant produced by biotechnology or the like while keeping it in a transparent container in a sterile state, or a cultivation method is widely known. In these technologies, how to maintain the plants in good condition and extend the viewing period is important. For that purpose, while maintaining the sterile condition, during the viewing period, oxygen gas and carbon dioxide gas related to the metabolic physiology of the plant are put in and out of the container, water from the medium is released, and the cloudiness in the container due to water is reduced, It is necessary to ensure that the viewing of plants is not disturbed. However, it is a seemingly contradictory request to prevent the invasion of microorganisms from the outside into the container and to have air permeability, which has been the largest technical issue to date.

Conventionally, for example, in the invention described in Japanese Patent Laid-Open No. 62-259514, a container is hermetically sealed in order to prevent the invasion of bacteria, and a hygroscopic agent is installed in the container. However, in this case, it is difficult to provide a long-term appreciation because the active period of the hygroscopic agent is limited. Also, JP-A-63-2
In the invention described in Japanese Patent No. 30021, as the container sealing structure, it is possible to utilize the flexibility of synthetic resin, cover with aluminum foil, seal with a filter, screw a lid and a container, and the like. However, it is only described functionally that bacteria, mold and the like do not enter the container from the outside, and that oxygen and carbon dioxide gas are sealed in and out naturally, and the specific means is not described at all.

According to the findings of the present inventors, as will be apparent from the comparative example described later, no matter how small a ventilation hole or a gap is provided between the lid and the container, the medium can be removed on the medium within 3 days. Mold will be generated. Further, even if it is called a filter, it is difficult to maintain a sterile condition for a long time with a normal filter medium or the like.

[0005]

SUMMARY OF THE INVENTION It is an object of the present invention to provide an aseptic plant sterile capsule which is free from the above-mentioned problems and is breathable and free from the invasion of microorganisms.

Means for Solving the Problems That is, the ornamental plant sterile capsule of the present invention which has achieved the above-mentioned object is an ornamental plant consisting of a transparent heat-resistant capsule container having a medium and a sterile plant placed in the medium. A sterile plant capsule, characterized in that the capsule container has at least one vent hole sealed with an air vent film.

In the present invention, the air vent film has air permeability and prevents passage of microorganisms, and a porous membrane filter made of polytetrafluoroethylene is used. The thickness of the membrane filter is not particularly limited, but usually 1
It is preferably about 25 μm. The maximum operating temperature should be approximately 120 ° C or higher, but it can withstand high-pressure steam sterilization stably. Are preferred. Even if the pore size is within the above range, if the porosity exceeds 80%, various bacteria may enter depending on the viewing environment. Further, depending on the viewing environment, even if the air vent film is out of this range, it has a certain effect, but as will be apparent from the examples described later, it can be viewed while maintaining a good condition for a long period of 6 months or more. It is not possible. It is convenient to use an air vent film having an acrylic adhesive layer on one side.

Next, embodiments of the capsule container of the present invention will be described in detail with reference to the drawings. In FIG. 1, the capsule container includes a base portion 1 and a transparent lid body 2 that is airtightly screwed into the base portion 1. The shape of the entire capsule container is not particularly limited, but it is preferably substantially cylindrical, and the plant can be seen through from this cylindrical portion. Also the base 1
Is provided with a culture medium chamber 1a, and further, a collar portion 1b on a funnel is provided concentrically outside the culture medium chamber 1a.
It is desirable to limit the amount of dead plant leaves to the collar part 1b to prevent the culture medium from being contaminated.
The material of the container may be glass, polypropylene, polyethylene or the like as long as it can withstand a high temperature sterilization treatment at about 120 ° C. for about 20 minutes, but a polycarbonate resin is most preferable in practical use. It can be colored if necessary, but the part where the ornamental plant is seen through must be transparent. In the present invention, a vent hole 3 is formed in the upper portion of the lid body 2, and the vent hole 3 is closed by the air vent film 4 as described above. The size of the vent hole 3 cannot be specified because it varies depending on the volume of the capsule container, the type of ornamental plant, and the like, but if it is too small, about 2 to 6 mm is necessary because the dissipation of water is insufficient. 6 is an ornamental sterile plant.

As the medium to be filled in the capsule container of the present invention, either a liquid medium or a solid medium can be used, but from the viewpoint of stability, a solid medium using colorless or colored agar or gellan gum is preferable. Gellan gum is particularly preferable from the viewpoint of transparency. For this medium, for example, Murashige-Skoog (MS) medium widely used for plant tissue culture is suitable. The medium composition basically has inorganic salts and carbon sources as main components, but amino acids, vitamins, plant hormones and the like can be included as trace components. Inorganic salts include ammonium nitrate, potassium nitrate, calcium chloride, manganese sulfate, potassium dihydrogen phosphate, disodium ethylenediaminetetraacetate, gnesium sulfate, iron sulfate, boric acid, potassium iodide, cobalt chloride, sodium molybdate, copper sulfate, sulfuric acid. A mixture such as zinc is used. Saccharose is generally used as a carbon source. As amino acids and vitamins, nicotinic acid, pyridoxine hydrochloride, thiamine hydrochloride,
Myo-inositol, glycine, etc. can be used. Naphthalene acetic acid (NAA) and benzylaminopurine (BA) are used as plant hormone agents.

Ornamental sterile plants include seeds, stems,
Shoot apex, germ part, root tip part, bulb part, root part, bulb germ part,
A part of the plant tissue such as flower bud, cambium, leaf and the like is extracted and tissue culture is used. Alternatively, the seed may be directly planted in a medium and aseptically grown may be transplanted.
As the type of plant, various plants such as ornamental plants and woody plants can be used. For example, rose, benjamin, coffee, Tsuwabuki, Hamayu, clover, etc. can be mentioned. In the present invention, the shoot apex of Tsuwabuki or the spotted Tsuwabuki, which has never been used so far, has been extracted and tissue-cultured seedlings have been successfully used to prepare an ornamental plant. In case of Tsuwabuki,
When seedlings grow and grow in full capsule containers, if they have chlorophyll, cut the stems from the roots and wash the roots well, avoid direct sunlight and plant them in trees or flower pots and grow them in the open field. You can do it, so there is no waste.

The aseptic plant sterile capsule of the present invention can be produced by the following method. The following seven types of stock solutions are prepared in advance for MS basic medium and stored in a refrigerator or a freezer. MS-0 Ammonium nitrate Potassium nitrate MS-1 Potassium dihydrogen phosphate Boric acid Manganese sulfate MS-2 Calcium chloride MS-3 Magnesium sulfate MS-4 Ethylenediaminetetraacetic acid disodium MS-5 Myoinositol (frozen) Glycine Nicotinic acid Thiamine pyridoxine hydrochloride MS -6 Potassium iodide Sodium molybdate Copper sulphate Cobalt chloride Zinc sulphate

Then, sucrose is added to these stock solutions and dissolved in deionized water to obtain MS basic medium (liquid). To this MS basic medium, an appropriate amount of plant hormones such as naphthalene acetic acid (NAA) or benzylaminopurine (BA) and agar or gellan gum are added according to the purpose, and the mixture is subjected to a steamer or the like to prepare a shoot-top culture medium, a seedling culture medium, A solid medium such as a growth medium and a rooting medium is obtained. When the agar or gellan gum is completely dissolved, dispense into containers. The rooting medium is dispensed into the medium chamber 1a of the transparent capsule container made of polycarbonate resin shown in FIG. 1, the lid body 2 and the collar portion 1b of the base portion 1 are screwed together, and silicon packing (not shown) is used. Etc. to be airtightly integrated. An air vent film 4 having an acrylic adhesive on one surface is previously attached to the ventilation hole of the lid body 2. The capsule container is put into an autoclave and sterilized by autoclaving at 115 ° C. for 20 minutes. After sterilization, it is placed horizontally and allowed to cool to solidify the medium.

On the other hand, in ornamental plants such as Tsuwabuki, unnecessary parts such as stems, leaves and roots are cut off and immersed in 75% ethanol for about 10 to 15 seconds. Then, in a clean bench, disinfect the surface with sodium hypozincate (antiformin) for about 2 minutes, and rinse with sterilized water. Under the stereoscopic microscope, the obtained sterile plants are separated from the leaves one by one from the outside, the stem apex portion is confirmed and cut off with a scalpel, and the plant is placed in a test tube filled with the shoot apical culture medium and stored in a culture shelf. After the cells have been grown for about 2 months, the cells are split and transplanted into a growth medium filled in another large container and further grown. During the growth, if necessary, such as at night, the growth can be promoted by illumination of about 2000 lux at room temperature (about 25 ° C.).
The strain grown to a predetermined size is cut with a knife and placed on the rooting medium in the capsule container described above.

The ornamental plant of the present invention can be aseptically grown from seeds separately from the above-mentioned tissue culture. For example, in the case of coffee seedlings, soak coffee beans in about 75% ethanol for about 20 seconds in a clean bench and 1%
Disinfect the surface by immersing it in antiformin for about 3 minutes, and then rinse with sterilized water several times. Then, it is placed in a petri dish, and sterilized water is put thereinto and left to stand for about 12 to 24 hours. After being sufficiently sucked with water, the seedling culture medium is transplanted into a flask. Wait for rooting and germination for 2-3 months, and transplant to the rooting medium in the capsule container described above.

[0014]

In the aseptic plant sterile capsule of the present invention, oxygen, carbon dioxide and water can pass through, but the invasion of microorganisms can be blocked by the action of the special air vent film with the vent hole closed. This makes the ornamental plant last longer and prevents the inside of the capsule from fogging due to moisture.

[0015]

Example 1 A capsule container made of polycarbonate resin shown in FIG. 1 was used. Of container base 1 The height was 75 mm. As a result, the total volume of the medium chamber 1a excluding the volume is about 130. It was closed with an air vent film 4 having a system adhesive. As the air vent film, a polytetrafluoroethylene mitex membrane (trade name, Millipore) %Met. A rooting medium described below is dispensed into the medium chamber 1a of the capsule container, and the base 1 and the lid 2 are airtightly screwed together, and then autoclaved at 115 ° C. for 20 minutes under high pressure steam sterilization, Allowed to cool and solidify.

A pre-prepared stock solution for MS medium was prepared to have the following composition and dissolved in deionized water,
The pH was adjusted to 5.8 to obtain MS basic medium (liquid).

On the other hand, spotted Tsuwabuki was used as an ornamental plant. First, unnecessary portions were cut off and then immersed in 75% ethanol for 15 seconds. The surface was then disinfected with 1% antiformin for 2 minutes in a clean bench and rinsed with sterile water. Under a stereoscopic microscope, the leaves were cut off one by one, the stem apex was confirmed, and the leaves were excised with a scalpel. This was placed on the shoot apical culture medium in a test tube and stored on a culture shelf. As the shoot apical culture medium, a solid medium obtained by adding the following amounts of plant hormones and agar to the MS basic medium was used.

After 2 months, the slightly grown strain was transferred to a mayonnaise bottle containing 350 ml of a growth medium. The growth medium includes
A solid medium prepared by adding the following amounts of plant hormones and agar to the MS basic medium was used.

A strain grown for 2 months was cut out to about 1 to 3 mm at room temperature at room temperature under illumination of 2000 lux and transplanted to a rooting medium in a capsule container to obtain a sterile Tsubakiki capsule for ornamental use. . As the rooting medium, a solid medium in which the following amounts of plant hormones and gellan gum were added to MS basic medium was used.

[0020]

Example 2 The same as Example 1 except that was used.

[0021]

Example 3 The same as Example 1 except that

[0022]

[Example 4] 75% coffee beans in a clean bench
Immerse in ethanol for 20 seconds. The surface was then disinfected by immersion in 1% antiformin for 3 minutes. The sterilized beans were washed twice with sterilized water, placed in a petri dish, filled with sterilized water and left for 12 to 24 hours to absorb water sufficiently, and then transferred to a flask containing a seedling culture medium. As the seedling culture medium, a solid medium obtained by adding the following amounts of plant hormones and agar to the MS basic medium was used.

The seedlings, which had grown sufficiently for 3 months, were transplanted to a rooting medium in a capsule container to obtain an aseptic coffee sterile capsule. As the rooting medium, a solid medium in which the following amounts of plant hormones and gellan gum were added to MS basic medium was used.

[0024]

Comparative Example 1 The same as Example 1 except that a lid without air holes was used to hermetically screw the base of the container and the lid to form a completely sealed capsule container.

[0025]

[Comparative Example 2] The same as Example 1 except that a cap without a vent was used to loosen the screwing between the base of the container and the cap to form a capsule container with a physical gap.

The results of observing the capsules of the above Examples and Comparative Examples for 6 months are as follows.

[Table 1]

From the above results, the capsules of the examples using the air vent film of the present invention are free from mold and fogging and can be viewed for a long time. Especially air vent film And in Example 4, the atmosphere in the capsule is maintained for 6 months or more, and the plant actually grows to fill the capsule and last long until it does not serve the purpose of appreciation.

[0028]

INDUSTRIAL APPLICABILITY As described above, in the ornamental plant sterile capsule of the present invention, the following effects are obtained by using the air vent film, and plants can be safely viewed indoors for a long period of time. (1). Oxygen and carbon dioxide, which are involved in the metabolic physiology of plants, can freely flow in and out, allowing long-term survival. (2). The water that escapes from the medium can be dissipated, so that the inside of the capsule becomes cloudy and the see-through of plants is not hindered. (3). Bacteria and microorganisms floating in the atmosphere do not enter the capsule, and the sterility can be maintained, so that the capsule can be viewed for a long time.

[Brief description of drawings]

FIG. 1 is a side sectional view showing an example of an aseptic plant sterile capsule of the present invention.

[Explanation of symbols]

 1 ... Base 1a ... Medium chamber 1b ... Collar 2 ... Lid 3 ... Vent 4 ... Air vent film 5 ... Medium 6 ... Plant

Claims (9)

[Claims] 1. An aseptic plant sterile capsule comprising a transparent heat-resistant capsule container having a medium and a sterile plant placed in the medium, the capsule container having at least one vent hole sealed with an air vent film. An aseptic plant aseptic capsule comprising: 2. The aseptic plant sterile capsule according to claim 1, wherein the medium is agar or gellan gum containing a nutrient and a plant hormone. 3. The capsule container comprises a base portion and a substantially cylindrical transparent lid body airtightly coupled to the base portion, and the sterile plant is viewed through the lid body. Aseptic plant sterile capsules as described. 4. The ornamental plant according to claim 3, wherein the base has a medium chamber and a collar, and the lid is airtightly coupled to the collar so that the leaves of the sterile plant do not contaminate the medium. Aseptic capsule. 5. The ornamental plant sterile capsule according to claim 1, 3 or 4, wherein the capsule container is made of a polycarbonate resin. 6. The sterile capsule for ornamental plants according to claim 1, wherein the aseptic plant is a seedling of Tsuwabuki or spotted Tsuwabuki which has been tissue-cultured by extracting from a part of the tissue. 7. The sterile capsule for ornamental plants according to claim 1, wherein the sterile plant is a seedling of coffee aseptically grown from seed in a medium. 8. The air vent film is porous made of polytetrafluoroethylene. The aseptic plant sterile capsule according to claim 1. 9. The sterile plant ornamental capsule according to claim 1, wherein the ventilation holes have a diameter of about 2 to 6 mm.