JPH0766A - Moss ball and method for culturing the same - Google Patents

Abstract

PURPOSE:To obtain a dense moss ball having an arranged shape. CONSTITUTION:This method for culturing a moss ball comprises subjecting the plant tissues of a moss to a sterilization operation, culturing the moss to divide into young plants, and subsequently culturing the young plants, while revolution-shaking the young plants so as to roll. The dense moss ball has an arranged shape comprising a central layer in which the moss or its residues are radially arranged and a surface layer containing the plants as a main constituting element.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to marigot and its culturing method.

[0002]

2. Description of the Related Art Recently, the problems to be solved by the invention
Mosses are attracting attention for appreciating Japanese gardens and bonsai. The beauty of moss is not caused by flowers like flower plants, but is exhibited by a large number of individuals (communities). However, it is difficult to cultivate a large amount of moss due to landscaping and the like because there are many unknown parts in its physiological ecology, and it is the current situation that it is necessary to rely on a horticulturist skilled in the cultivation. .

By the way, "marigoke" is known as a moss designated as a natural monument of Japan. This Marigot is a group of moss that lives near the shores of Lake Akan, Lake Inawashiro, etc., extracted by the strong waves and swung by the waves of the shallow water, while gathering with the water plants and small pieces of garbage there. It has become a sphere or an ellipsoid.

[0004] As such moss which becomes Marigot in the natural world, there are representatives such as Hilohanosukigogoke and Kinusippogoke. The Marigots formed in such natural world are moss plants as described above. Since the body is entangled in a spherical shape with aquatic plants and small pieces of dust, etc., there are problems that the shape is distorted, the quality is rough, and many are soft, which are not suitable for viewing. Moreover, marigots are formed only when the balance of many environmental conditions is suitable, and their cultivation is difficult, and there is a problem that immediately after cultivation, new shoots grow upward and lose their shape. If well-formed and precise Marigots can be obtained, not only Japanese-style gardens and bonsai but also demand for decorations in tanks as an alternative to Marimo can be expected.

[0005] Therefore, an object of the present invention is to provide a marigo moss that is well-shaped and dense.

[0006] Another object of the present invention is to provide a method for easily producing a dense Marigot having a regular shape.

[0007]

As a result of earnest research in view of the above-mentioned object, the present inventors have conducted a sterilization operation on plant tissues of moss, and then cultured and differentiated into young plant bodies. The present invention has been found out that it is possible to easily produce a dense Marigot having a uniform shape by culturing while shaking the plant so that the plant rolls.

[0008] That is, the marigot of the present invention is composed of an almost spherical aggregate of moss, and has a central layer in which moss or its remnants are radially arranged and a plant as main constituent elements. And a surface layer.

Further, the method of the present invention for producing the above-mentioned Marigot is (a) after sterilizing a moss plant tissue,
It is characterized by culturing and differentiating into a young plant, and (b) shaking culture so that the young plant rolls.

The present invention will be described in detail below. [1] Marigots Marigots of the present invention basically consist of a central layer in which moss or its debris are radially arranged, and a surface layer whose main constituents are plants. Such Marigots are roughly classified into two depending on the configuration of the central layer (hereinafter, referred to as a first Marigot and a second Marigot depending on the difference in the configuration).

(1) The first moss, which is the first moss, is a moss that does not form a filamentous body in the culture medium and basically grows in a plant body, and is almost spherical by the culture method described below. It will be. Examples of such moss include moss, moss moss, moss moss, moss moss, moss moss, moss moss, moss moss, moss moss, moss moss, moss moss, and the like.

FIG. 1 schematically shows the structure of the first Marriage moss of the present invention comprising such moss. In FIG. 1, the boundaries between the layers are shown as clear lines for convenience of explanation.
In reality, there are no such boundaries between layers. The first Marigot mosquito 1 has a substantially spherical shape, and is composed of a central layer 2 in which moss or its debris are radially arranged, and a surface layer 3. In addition, in the present invention, the term “nearly spherical” means not only a true sphere but also a distorted sphere having a certain shape, an ellipse or an oval shaped rotating body, an egg shape, a shape such as a rugby ball,
Alternatively, these shapes include those that are slightly distorted.

The central layer 2 is composed of a temporary root, a plant, and / or the remains thereof as constituent elements, and is a layer in which tissues are arranged approximately radially from the central layer. In the central layer, the ratio of the temporary root (and / or its debris) and the plant body (and / or its debris) is such that the temporary root (and / or its debris) is about 10 to 25%, and the plant body (and / Or the remains) is about 75 to 95%.

When Marigoke grows to a diameter of about 5 cm or more, most of the tissue up to about 2 cm from the surface is alive, but most of the tissue deeper than that has already died and turned brown. In general, the moss tissues that have died and become brown in this way are called remnants in the present invention.

The surface layer 3 is a layer containing a plant as a main constituent element, and may contain a small amount of temporary roots.

The size (diameter) of the above-mentioned Marigot moss
Is about 1 to 10 cm, preferably about 2 to 7 cm.
If it is less than about 1 cm, it is not sufficiently spherical, and it is difficult to make it larger than about 10 cm. The size (radial length) of each layer is generally 0.2-1.
5 cm, especially 0.2-0.5 cm, surface layer 3
Is about 0.8 to 2.0 cm, preferably about 1.0 to 1.3 cm. For example, a diameter of about 5 cm (radius about 2.5 c
m), the radial length of the central layer 2 is about 0.5 to
At 1.0 cm, the surface layer 3 becomes about 1.5 to 2.0 cm.

[0017] In such a first marigot, first, one or more individuals of moss are entangled with each other to form a nucleus, from which a plant grows, and from the plant, a temporary root and a plant are further formed. By repeating the germination of the body and growing radially outward, the central layer 2
Is obtained. That is, in the Marigot of the present invention, the outermost layer is always the plant body layer (surface layer), and the plant body layer (surface layer) is further outside by germinating the roots and the plant body from the plant body. It is presumed that the portion that had been transferred and was originally the plant body layer (surface layer) will gradually grow by assimilating with the central layer 2.

(2) Second Marriage Moss The second moss that serves as the second moss of the present invention grows in a culture solution on a filamentous body and a plant body, and grows in a substantially spherical shape by a culture method described later. To do. Examples of such moss include moss, moss, moss, moss, moss, moss, moss, moss, and moss.

FIG. 2 schematically shows the structure of the second Marigot which consists of such moss. In addition, in FIG.
The boundaries of the layers are shown as clear lines for convenience, as in the case of the first description of Marigot. The second Marigot 11 has a substantially spherical shape, and is composed of a center layer 12 in which moss or its debris are radially arranged, and a surface layer 13.

The central layer 12 is composed of a raw thread, a root, a plant, and / or the remains thereof as constituents. In the center layer 12, the ratio of the raw thread (and / or its remnants) to the temporary roots (and / or its remnants) and the plant body (and / or its remnants) is calculated as follows: ) Is 50
~ 70%, and temporary roots (and / or its debris) 20 ~
The plant body (and / or its remains) is about 10% at about 30%.
It is about 20%.

The surface layer 13 is a layer containing a plant as a main constituent element, and may be slightly mixed with a temporary root and a raw thread.

The size (diameter) of the above-mentioned Marigot 11 is about 0.5 to 10 cm, preferably about 1 to 6 cm. If it is less than about 0.5 cm, it will not be sufficiently spherical, and it will be difficult to make it larger than about 10 cm. In addition,
The size (radial length) of each layer is generally 0.4 to 2.0 cm, particularly 1.0 to 1.3 cm for the central layer 12, and 0.1 to 1.0 cm for the surface layer 13. Degree, especially 0.2-
It is preferably about 0.5 cm. For example, about 5 cm in diameter
When the radius is about 2.5 cm, the radial length is about 2.0 to 2.3 cm for the central layer 12 and about 0.2 to 0.5 cm for the surface layer 13.
Becomes

In such a second Marigot, first, one or two or more individuals of moss are entangled with each other to form a nucleus (central layer 12), from which a raw thread grows, and the raw thread thereof is formed. It is obtained by forming the growth layer 12 by repeating germination of the plant body from the body and further germination of the protophyte, the plant body and the temporary root from the plant body. That is, the outermost part of the second marigot is always a plant body layer (surface layer), and the protozoa, the plant body, and the roots germinate from the plant body, so that the plant is further outside. The body layer (surface layer) 13 is formed, and the initial plant body layer (surface layer) 13 is formed.
Is supposed to grow by assimilating with the central layer 12. If the protozoa, the plant, and the roots are further germinated from the plant forming the surface layer, the surface layer should include not only the plant but also the protozoa and the plant. Since most of the roots grow in the central direction and most of the protozoa along the surface, almost only the plant stands in the outer direction. Therefore, judging from the surface of Marigot moss, the surface layer mainly contains plants.

[2] Culturing Method Next, a method for cultivating the above-mentioned Marigot mushroom of the present invention will be described.

(1) Collection of moss First, moss plants, spores, etc. are collected from the mountains. As the moss to be collected, desired ones can be used among the above-mentioned first and second moss to be the moss. The moss plant tissues to be cultured include moss gametophytes, cell debris, spores, asexual buds, rhizomes,
Examples include temporary roots and raw thread bodies. In addition, callus derived from each plant tissue can also be used.

(2) Medium The moss plant tissue thus obtained is used to cultivate Marigot, but the medium used in the present invention is not particularly limited, and MS medium (Murashige-Skoog medium) is used. ) And various MS modified mediums described below (Murashige-Skoog
Deformation medium), etc. are appropriately selected according to the moss to be used, and if necessary, a plant growth regulator (kinetin,
It is possible to use those to which benzyladenine, indole-3-acetic acid, 2,4-dichlorophenoxyacetic acid (2,4-D), etc. are added.

As the MS modified medium, the following So is used.
l. 1 to 3 (solvent is distilled water), saccharides such as glucose and vitamins, organic acids, inorganic acids and salts thereof, and the like, and a combination of distilled water, for example, NA-MS shown below. Medium, AS-MS medium, ANA-MS medium, AAS-MS medium, MSF medium, and these
Examples include those diluted 10-fold.

Sol. 1 Calcium chloride dihydrate 4,400 mg / liter magnesium sulfate heptahydrate 3,700 mg / l manganese sulfate 4-5 hydrate 223 mg / liter boric acid 62 mg / l zinc sulfate heptahydrate 107 mg / liter Sodium molybdate dihydrate 2.5 mg / liter Copper sulfate pentahydrate 0.25 mg / liter Cobalt chloride hexahydrate 0.25 mg / liter Potassium iodide 8.3 mg / liter

Sol. Monopotassium diphosphate 42.5 mg / l

Sol. 3 myo-inositol 10,000 mg / liter Nicotinic acid 50 mg / liter Pyridoxine hydrochloride 50 mg / liter Thiamine hydrochloride 10 mg / liter

Composition of various MS modified media NA-MS media Sol. 1 100 ml / liter Sol. 24 ml / liter Sol. 3 10 ml / liter Glucose 20 g / liter Potassium nitrate 4 g / liter Ethylenediaminetetraacetic acid / disodium 47 mg / liter Iron sulfate heptahydrate 27.8 mg / liter pH About 5.7

AS-MS medium Sol. 1 100 ml / liter Sol. 24 ml / liter Sol. 3 10 ml / liter Glucose 20 g / liter Ethylenediaminetetraacetic acid / sodium 47 mg / liter Iron sulfate heptahydrate 27.8 mg / liter Ammonium chloride 44 mM Sodium succinate or sodium fumarate 33 mM pH About 5.8

ANA-MS medium Sol. 1 100 ml / liter Sol. 24 ml / liter Potassium nitrate 4 g / liter Ethylenediaminetetraacetic acid / disodium 47 mg / liter Iron sulfate heptahydrate 27.8 mg / liter

AAS-MS medium Sol. 1 100 ml / liter Sol. 24 ml / liter Ethylenediaminetetraacetic acid / disodium 47 mg / liter Iron sulfate heptahydrate 27.8 mg / liter Ammonium chloride 44 mM Sodium succinate or sodium fumarate 33 mM pH About 5.8

MSF medium Sol. 1 10 ml / liter Sol. 2 0.4 ml / liter Sol. 3 1 ml / liter Glucose 2 g / liter Potassium nitrate 0.4 g / liter Ethylenediaminetetraacetic acid / 2 sodium 4.7 mg / liter Iron sulfate heptahydrate 2.78 mg / liter Ammonium chloride 20 mM Sodium succinate or sodium fumarate 15 mM pH About 5.8

(3) Cultivation The sterilized plant tissue is subjected to a sterilization operation using such a medium, and the sterilized plant thus obtained is shake-cultured under predetermined conditions to obtain the present invention. You can get Marigot of.

Hereinafter, the method for producing marigota of the present invention will be described in 1 /
5 (means diluted 5 times; the same applies hereinafter) NA-
The case where MS medium is used will be described step by step.

(3-1) Aseptic Operation First, the collected bryophyte plant tissues (gametophyte, cell debris, spores, asexual buds, rhizomes, rhizomes, protozoa, etc.) are cut into about 1 to 10 mm. Then, after thoroughly cleaning with an ultrasonic cleaner or the like, sterilization treatment is performed.

The sterilization treatment is carried out by using polyoxyethylene sorbitan monolaurate (for example, trade name: Twee
n 20), polyoxyethylene sorbitan monooleate
(For example, wash with nonionic surfactant such as Tween 80) or cationic surfactant solution (concentration 0.1 to 0.5% by volume) such as benzalkonium chloride for 40 to 60 seconds. It can be performed by sterilizing with 0.5 to 1% by volume of sodium hypochlorite solution and immediately followed by thorough washing with sterile distilled water.

Next, the sterilized moss tissue was
Cultured on 5NA-MS medium (solid: solidified with agar, gellite, etc.) while irradiating with light of about 1000 to 3000 lux at 20 to 25 ° C, and a site contaminated with other fungi After culturing, the culturing is continued again until the site contaminated with other fungi and the like disappears, whereby a sterilized moss plant tissue can be obtained.

(3-2) Marigotization and Culture The thus sterilized moss tissue is ⅕N
It is placed in an A-MS medium (liquid) and shake-cultured for 6 months or longer in a state in which the moss tissue rolls. The culture solution should be replaced after about 1 month. As a method of shaking culture,
There is no particular limitation as long as the sterilized tissue rolls, and examples thereof include a swirling method and a method of rolling moss by bubbling, and a swirling shaking culture is particularly preferable. .

In the case of orbital shaking culture, the orbital speed is
It is preferably 100 to 140 rpm, particularly 110 to 120 rpm. When the swirling speed is less than 100 rpm, the sterilized moss tissues do not roll sufficiently, so that a group of moss does not develop in a spherical shape.On the other hand, when the swirling speed exceeds 140 rpm, the container is crushed by centrifugal force. The moss tissue is pressed against the outer wall and does not roll.

Culture conditions other than the above include 20 to
It is preferable to irradiate at a temperature of 25 ° C. and a light amount of about 8,000 to 10,000 lux.

The culture vessel to be used is not particularly limited, and an appropriate size can be used according to the desired size of the moss, but the moss tissue can be efficiently transferred. For dynamic culture, the bottom diameter should be 10-15
It is preferable to use an Erlenmeyer flask having a size of about cm.

By culturing the sterilized moss tissue in this manner for about 3 months, the seedlings of the present invention, which will be the core of the moss, can be obtained. However, the seedlings
At this stage, it does not yet have the structure as described above. By culturing the seedlings with shaking for 3 to 5 months, one individual of the seedlings or an aggregate of 2 to 5 individuals grew to have a diameter of about 3
You can get cm of moss. The growth process at this time is different between the first marigot and the second marigot as described above.

In the step of transforming the seedlings into moss, the porous bodies such as porous ceramics having a diameter of about 3 to 10 mm are cultivated at a rate of 1 to 5 to 10 seedlings. It can be mixed in the liquid. By mixing the porous body, the moss roots and the like are easily entangled in the pores, and the nucleus is efficiently formed.

After that, since this Marriage moss continues to grow by about 5 to 10 mm every 1 to 2 months by continuing the shaking culture, the shaking culture should be continued under the same conditions until the desired size is reached. Good. The upper limit of the size of the moss, which depends on the size of the container to be cultivated, is about 7 cm or less in diameter as described above.

In this culturing step, the amount of the culture medium may be such that at least about half of the moss is submerged, and it is preferable to replace the culture broth approximately every one month.

Thus, the Marriage moss of the present invention can be cultivated, but the culturing method of the present invention can be applied in various ways by culturing moss plant tissue while rolling. For example, the medium used is 1/5 NA-M
Not limited to S medium, various media suitable for culturing the moss can be used. Further, the degree of enrichment of carbon dioxide in the medium can be variously changed depending on the moss to be cultured and the environment.

[0050]

The present invention will be described in more detail with reference to the following specific examples.

Example 1 A moss moss was collected, and a plant having relatively little damage such as worm eating of the moss was separated and the plant was about 0.5 cm.
It was cut into pieces and washed well with an ultrasonic cleaner. Subsequently, it was washed with a nonionic surfactant solution (concentration: 0.5% by volume) for 40 seconds, then with a 1.0% by volume sodium hypochlorite solution for 40 seconds, and then thoroughly washed with sterile water. did.

One of the cut pieces thus washed is
Culture was performed on a / 5NA-MS solid medium (solidified with about 0.8% agar) while irradiating with a light amount of 2000 lux at 25 ° C. to remove the site contaminated with other fungi. After that, the operation of culturing again was repeated three times to obtain a sterilized plant.

The sterilized plant thus obtained was placed in an Erlenmeyer flask having a diameter of 11 cm on the bottom surface containing a sterilized ⅕NA-MS medium (liquid), and cultivated shaking culture with light control function. Machine (ROTARY SHAKE made by Sanki Seiki Co., Ltd.
(R Model GR-S) at 25 ° C, irradiating with light of 8000lux, and culturing for 60 days while rotating at 115 rpm / min to promote differentiation into seedlings that will be the core of Marigotoke. did.

Ten of these seedlings were placed in a 1/5 NA-MS medium and shake-cultured under the same conditions for 90 days by swirling to obtain Marigotoke having a diameter of about 2 cm.

Thereafter, the culture was continued for 90 days under the same conditions while exchanging the culture medium every 30 days to obtain Marigots with a diameter of about 4 cm. The thus obtained Marriage moss was cut, and its tissue was disentangled to examine the constituent elements. The region within a radius of about 1.5 cm from the center had the roots, plants, and their remnants as constituent elements. It was confirmed that the surface layer was the central layer, and the area up to about 0.5 cm from the surface was the surface layer mainly composed of the plant body.

Example 2 Eggs were collected, and the plants with relatively little damage such as worm eating of these eggs were separated, and the plants were separated by about 0.5.
It was cut to about cm and washed well with an ultrasonic cleaner. Then, add 40 with nonionic surfactant solution (concentration 0.5% by volume).
It was washed for 10 seconds, then washed with 1.0% by volume of sodium hypochlorite solution for 40 seconds, and then thoroughly washed with sterile water.

One of the cut pieces thus washed is
Culture was performed on a / 5NA-MS solid medium (solidified with about 0.8% agar) while irradiating with a light amount of 2000 lux at 25 ° C. to remove the site contaminated with other fungi. After that, the operation of culturing again was repeated three times to obtain a sterilized plant.

The sterilized plant thus obtained was placed in an Erlenmeyer flask having a bottom diameter of 11 cm and containing a sterilized ⅕NA-MS medium (liquid), and cultivated shaking culture with light control function. Machine (ROTARY SHAKE made by Sanki Seiki Co., Ltd.
(R Model GR-S) at 25 ° C, irradiating with light of 8000lux, and culturing for 60 days while rotating at 115 rpm / min to promote differentiation into seedlings that will be the core of Marigotoke. did.

10 seedlings were treated with 1/5 NA-MS
The mixture was placed in a medium and cultivated with shaking under the same conditions for 90 days in a swirling manner to obtain Marigots with a diameter of about 1 cm.

Thereafter, the culture was continued under the same conditions for 90 days while exchanging the culture medium every 30 days, and the diameter was about 2.5 cm.
I got Marigotoke. When the Marigot moss obtained in this way was cut, its tissue was disentangled and the constituent elements were examined,
The area within a radius of about 1.0 cm from the center is the central layer that consists of protofilaments, temporary roots, plants, and their debris, and the area from the surface to about 0.3 cm contains plant bodies. It was confirmed that the surface layer was the main component.

[0061]

As described above in detail, in the present invention,
After sterilizing the plant tissue of moss, it is cultivated and differentiated into seedlings, which are cultivated while swirling so that the seedlings roll. You can get precise Marigot.

The above-described Marigot mushroom of the present invention is suitable not only for Japanese gardens and bonsai trees, but also for decoration in aquariums as a substitute for marimo.

[Brief description of drawings]

FIG. 1 is a schematic diagram showing the structure of an example of a marigot mushroom of the present invention.

FIG. 2 is a schematic view showing the structure of another example of Marigot moss of the present invention.

[Explanation of symbols]

 1 ... 1st Marigot 2 ... Central layer 3 ... Surface layer 11 ... 2nd Marigot 12 ... Central layer 13 ... Surface layer

Claims (4)

[Claims] 1. A moss consisting of an almost spherical aggregate of moss, comprising a central layer in which moss or its remnants are radially arranged, and a surface layer whose main constituents are plant bodies. Marigot which is characterized by. 2. The Marriage mosquito according to claim 1, wherein the central layer comprises a temporary root, a plant, and / or a remnant thereof. 3. The Marriage mosquito according to claim 1, wherein the central layer is composed of a raw thread, a root, a plant, and / or a remnant thereof. 4. The method for cultivating Marigots according to any one of claims 1 to 3, wherein (a) the plant tissue of moss is sterilized and then cultured to differentiate into a young plant. , (B)
A method for cultivating Marigotoke, which comprises shaking culture so that the seedlings roll.