JP2005046059A - Method for field artificial proliferation of foliar thallus and material utilizing foliar thallus proliferated by the method for artificial proliferation therefor - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To solve problems that a thallus extremely slowly grows and has difficulty in artificial culture, to readily redifferentiate a thallus from a fungus and an alga in vitro, to make the redifferentiated thallus synthesize the same chemical component as that of a natural substance, to take out the thallus from in vitro and to grow the thallus in the same manner in a field. <P>SOLUTION: A piece of the thallus is cut out from field Parmelia tinctorum, transplanted on the surface of a base article, covered with a nylon mesh and fixed, a hemispheric projection is formed from around an alga layer in the thallus, enlarged in an Isidea state, fused with an adjacent lobule in an Isidea state, the lobule in the Isidea state is enlarged to a palmar state in a development stage, a hypomembraneous layer rhizomorph produced in the formation of the lobule is fixed to the base article and the thallus is proliferated. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

  The present invention relates to a method for artificially proliferating foliate lichens (for example, genus Mushrooms, turnips, etc.) and a thing using a foliate lichen proliferated by this artificial propagation method.

  As is well known, many lichens can live in harsh environments such as Antarctica, Takayama, and the desert. However, the resistance to environmental changes due to air pollution and urbanization is very low, and even a slight environmental change kills it easily, or the number of individuals is extremely reduced and its growth is suppressed. In consideration of this characteristic, it is also used as an indicator plant for air pollution.

  And in the actual situation, the main factor driving this type of lichen to extinction is an increase in the amount of sulfur dioxide in the atmosphere. For example, if the concentration of sulfur dioxide exceeds 0.02 ppm in Tokyo, Can no longer grow. Other causes of death include acid rain, dryness created by an artificial environment, and logging of natural forests.

  For reference, the plant version Red List (conventional technology) published by the Environment Agency in 1997 lists 82 species of lichens that may be extinct among domestic lichens. That is, three species that are extinct (species that are considered extinct in Japan), 22 species that are endangered (species that are difficult to survive if the pressure factors that brought about the current situation continue to act), and II species that are endangered ( If the pressure factors that brought about the current situation continue to work, 23 species that are likely to move to the “Endangered Class I” rank in the near future), semi-endangered (currently low extinction risk, but the habitat conditions Depending on the change, it is considered to be 17 species that have elements that move to higher ranks as “endangered”, and 17 species that lack information (species that are considered to be rare but lack sufficient information to evaluate). .

  The above situation and lichens are extremely slow in growth and difficult to cultivate artificially. Therefore, a new artificial cultivation method has been strongly desired in the industry. Therefore, the present applicant established a method for easily redifferentiating lichens from fungi and algae in a test tube, and filed the method as a patent (Japanese Patent Laid-Open No. 4-63528). It was clarified that the same chemical components are synthesized. However, it was impossible to take out the lichen body in the test tube out of the test tube and grow it on the bark or the like as in the field.

  And this kind of pine with ume tree, moss pine on plum branches, moss plum, etc. have been prized in Japan since ancient times. However, until now, attempts have been made to artificially produce koke pine and koke plum, but there has been no successful example. However, a group of Prof. Bando and others at Kinki University cut out the Japanese amber mushrooms, cultured them in plastic bags, measured their diameters on a regular basis, and found optimal culture conditions, but they did not confirm regeneration.

JP-A-4-63528

  In the above-mentioned literature, a cell population composed of undifferentiated fungi and algae is obtained from a part of a differentiated lichen body and cultured under specific conditions. The redifferentiated lichen body has the same chemistry as natural. This is the stage where it has been clarified that the components are synthesized, and it is the content that has been cultured in a test tube. Therefore, it was cultured under the condition of aseptic condition in a test tube, and it was not taken out of the test tube and grown on bark or the like in the same manner as in the field. In addition, the culture was performed under the condition of shielding bacteria and molds. Therefore, it is essentially different from field culture.

  Therefore, the present invention is cultivating (living) the lichen body in the field under the condition that there is no air pollution (clean air environment), and cultivating the lichen body in the field. It is intended to be used as a bioindicator for human growth and human growth.

  In the invention of claim 1, for example, the experiment site is an area where many large leaf-like lichens are seen in the vicinity, and the cleanliness of the atmosphere is that there are few substances that are a source of pollution such as sulfur dioxide in the atmosphere. Possible areas and climatic conditions are those in which the average annual temperature is relatively warm and the precipitation amount exceeds 2000 mm.In these areas, the lichens are cultured in the field, and the lichens are It is intended to ensure supply to the desired industry, or for human growth and use as a biological indicator. In addition, it is intended to easily induce the redifferentiation of Amaranthus by selecting such a place and using a nylon mesh and attaching it to the base.

Claim 1 is: (1) A piece of lichen cut out from the peripheral part of a large Umenochoke growing in the field,
(2) The cut lichen body is transplanted to the surface of a tree species such as cedar, plum, pine, or the base material such as stone,
(3) The transplanted lichen body is covered with a mesh material such as nylon mesh or vinyl mesh for preventing the dropout, and the lichen body is fastened to the base with fasteners such as staples, nails, pins, adhesives,
(4) By the formation of a hemispherical protrusion from the algal layer in the fastened lichen body, and by the expansion of this protrusion, an ideal shape expansion, and by this expansion, by fusion of the adjacent small fragments of the ideal shape , The developmental stage of expanding the isidea-shaped fragment into a palm shape;
(5) Apricot mushroom characterized by the establishment of the base and the proliferation of lichens through the hypodermis false roots that occur at the time when these expanded small fragments (fissures) are formed It is an artificial propagation method in the field.

  Further, the invention of claim 2 provides a method for artificially proliferating apricot moss in the field optimal for achieving the invention of claim 1.

Claim 2 is: (1) A square lichen body having a length of 4 mm to 10 mm and a weight of 2 mg to 8 mg is cut out from a peripheral portion of a large leaf-like lichen growing in the field,
(2) Transplant the cut-out square lichen onto the surface of a tree species such as cedar, plum, pine, or a base such as stone,
(3) The transplanted lichen body is covered with a mesh material such as nylon mesh or vinyl mesh for preventing the dropout, and the lichen body is fastened to the base with fasteners such as staples, nails, pins, adhesives,
(4) Formation of hemispherical protrusions from the algae layer in the clamped lichen body and the expansion of the protrusions by the growth of the protrusions. A developmental stage in which the icidal fragment is expanded into a palm by fusion;
(5) At the stage where this medullary layer is covered with the lower skin layer, and the medullary layer is covered by the lower skin layer through the inferior hypodermis false root that occurs at the time when this expanded small fragment (split-like one) is formed. After this hypodermis layer is formed with the mycelium of the medulla layer,
(6) A method for artificially proliferating the Japanese apricot, characterized by the development of the lower skin layer and the proliferation of lichens.

  The invention of claim 3 is based on the fact that pine with this kind of plum tree, pine pine on the branch of plum, moss plum, etc. have been prized since ancient times in Japan. It is intended to provide bonsai with good value and good sense by making effective use of koke plum.

  Claim 3 uses the agate mushrooms grown by the artificial propagation method, characterized in that the agate mushrooms produced by the outdoor artificial propagation method of claim 1 or claim 2 are vegetated on bonsai soil or trees. It is a thing.

  The invention of claim 4 is intended to provide a biological indicator that can easily know the state of air pollution in the area where they live in the future. Also, you can stick this small Umbrella fragment on a suitable board, hang it on a tree branch such as cherry blossoms in the school, and let it stand for several months, so you can know the state of air pollution from the state of Umno Moke, so elementary and junior high school children, It is intended to be usable as an effective material for environmental education for students, and to be commercially available as this biological indicator.

  Claim 4 uses the agate mushrooms grown by the artificial growth method, characterized in that the agate mushrooms produced by the outdoor artificial growth method of claim 1 or claim 2 are used as a biological indicator of air pollution. It is a thing.

  The invention of claim 1 is as follows: (1) A piece of lichen body is cut out from the peripheral part of a large Umenochoke growing in the field; (2) The cut lichen body is a tree species such as cedar, plum, pine, or stone (3) Cover the lichen body with a mesh material to prevent falling off, and fasten the lichen body to the base with fasteners. (4) From the algal layer in the lichen body The formation of a hemispherical protrusion, the expansion of the protrusion, and the enlargement of the protrusion, and the enlargement of the protrusion, and the fusion of the adjacent small fragments of the ideal (the organ called fibroblasts) expands the small fragment of the ideal shape into the palm of the hand (5) Developmental stage (morphogenesis), (5) Artificial propagation in the field of the umbilical moss, which establishes the base through the hypodermis false root that occurs at the time when the expanded small fragments are formed, and promotes the growth of lichens Is the method.

  Therefore, for example, claim 1 is that, for example, the experiment site is an area where many large leaf-like lichens are found in the vicinity, and the cleanliness of the atmosphere is that there are few substances such as sulfur dioxide in the atmosphere. Possible regions, and also the climatic conditions, the annual average temperature is relatively warm, the region where the precipitation is about 1000 mm to 3000 mm is specified, and the lichens are cultured in the field in this region, It has features such as being able to reliably supply to the industry that demands lichens, growing humans, and being able to use it as a biological indicator. In addition, there is a feature that it is possible to easily induce the re-differentiation of the apricot mushroom by selecting such a place and using a nylon mesh and a method of fastening to a base.

  The invention of claim 2 is as follows: (1) A square lichen body having a length of 7 mm and a weight of 4 mg is cut out from a peripheral portion of a large genus Umenogoke growing in the field, (2) a square lichen body, (3) Cover the lichen body with a mesh material to prevent it from falling off, and fasten the lichen body to the base with fasteners. (4) The formation of hemispherical protrusions from around the algal layer in the square lichen body, the expansion of the ideal shape by the growth of the protrusion, and the fusion of the adjacent small fragments of the ideal shape, (5) Stages in which the medulla layer is covered by the lower skin layer, and (5) the root is fixed to the base via the false root of the lower skin layer, which is generated when the enlarged small fragment is formed. Then, after the lower skin layer is formed with mycelium of the medulla layer, (6 And development of the lower skin layers, or an artificial growth method in the field of Parmeliaceae achieving growth of lichen body.

  Therefore, Claim 2 can provide a method for artificially proliferating the ume tree in the field that is optimal for achieving the invention of Claim 1.

  The invention of claim 3 is a genus mushroom grown by an artificial propagation method, characterized in that the ume tree produced by the method for artificial propagation in the field of claim 1 or claim 2 is vegetated on bonsai soil or trees. It is a thing using.

  Therefore, claim 3 claims that this kind of pine tree with ume tree mushrooms, pine tree pine tree attached to a branch of plum, moss ume tree, etc. have been prized since ancient times in Japan. By making effective use of moss plums, we can provide bonsai with an excellent sense of product value.

  The invention of claim 4 uses the ume tree produced by the field artificial growth method according to claim 1 or claim 2 as a biological indicator of air pollution, characterized in that the ume tree grown by the artificial growth method is used. It is a thing using.

  Therefore, claim 4 can provide a biometric indicator that can easily know the state of air pollution in the area where they live in the future. Also, you can stick this small Umbrella fragment on a suitable board, hang it on a tree branch such as cherry blossoms in the school, and let it stand for several months, so you can know the state of air pollution from the state of Umno Moke, so elementary and junior high school children, It is characterized by the fact that it can be used as an effective material for environmental education for students, and can be marketed as a biological indicator.

Hereinafter, embodiments (examples) of the present invention will be described.
1. Example 1
1-1 Setting the experimental conditions (1) Collection location and experiment location Collection location: Kimitsu City, Chiba Prefecture Experiment location: Same as above

(2) Method ・ Cut out a square lichen body (4 mg in weight) with a length of 7 mm from the periphery of a large Umenochoke growing in the field.
・ Base tree species: To prevent cedar and lichens from falling off, put nylon mesh and staple the base. This work ensures light irradiation (which is an absolute condition) and prevents scattering.
・ As an investigation method of the results, external forms on both sides were taken and displayed with a stereomicroscope.
The internal form was taken with an optical microscope and displayed in a drawing.
A fine external form was taken with a scanning electron microscope and displayed in a drawing.
Displayed using the section preparation method (frozen microtome) during internal morphology observation.

1-2 Results (1) Ecological photographs are shown in Figs.
Figure 1 shows the date of transplantation on May 5, 2001.
Fig. 2 is about 4 months after the start of transplantation on September 27, 2001.
FIG. 3 is about 15 months (1 year and 3 months) after the start of transplantation on August 3, 2002.
In the above figure, FIG. 2 shows no small fragments in the periphery, but FIG. 3 clearly shows many small pieces formed from the periphery.

(2) The external form was photographed with a microscope and displayed in FIGS.
FIG. 4 shows the surface in a drawing.
FIG. 5 shows the back side of the drawing.

(3) Photographed with a scanning electron microscope and displayed in FIG. 6 to FIG. 11, but in general, the cross-section of the lichen body shows a part where the development stage has advanced and a slow part at the same time. You can follow the steps. A specific example will be described below.
In FIG. 6, the surface of the differentiated lichen was photographed with a scanning electron microscope and displayed in a drawing.
In FIG. 7, the back side of the differentiated lichen was photographed with a scanning electron microscope and displayed in a drawing. False roots can be observed.
In FIG. 8, the newly formed buds on the surface of the transplanted lichen (old) were photographed with a scanning electron microscope and displayed in a drawing.
FIG. 9 shows a very early lichen formed on the cut surface, and a state in which the external form resembles rupture was photographed with a scanning electron microscope and displayed in a drawing.
In FIG. 10, the lichen body in a slightly grown state was photographed with a scanning electron microscope and displayed in a drawing.
In FIG. 11, a lichen fully developed in the shape of a palm is photographed with a scanning electron microscope and displayed in a drawing.
Since the contents of the generation stage illustrated above have been examined, the results are described below.
a: A hemispherical protrusion is formed around the algal layer. Protrusions grow and are very similar in shape to the ideas formed on the lichen surface.
b: This isidea-like thing gradually grows and develops in the shape of a palm. Adjacent small fragments are expected to fuse and become larger. The reason is presumed from the fact that the number of isidea formed and the number of small pieces are small when compared under the same length.
c: The mycelium of the medulla layer is still exposed at the time when isidea is formed, but this medulla layer is eventually covered with the lower skin layer (judged by brown), and the formation of the lower skin layer is exposed Looks like it can be made from mycelium.

(4) Internal form FIG. 12 shows the state of the initial stage of occurrence (when forming an ideal cylindrical lichen) taken with a microscope and displayed in a drawing. The outline is as follows.
・ The epithelial layer is continuous.
・ Algae layer is not continuous.
-The inferior layer does not develop in the medulla layer (the medullary hyphae are bare).
I wrote this in the drawing.
FIG. 13 shows the state of late development (at the time of the formation of a fallen egg-shaped fragment) taken with a microscope and displayed in a drawing. The outline is as follows.
・ The epithelial layer and algae layer are continuous with the old and new lichens.
・ In the new lichen body, the development of the medulla layer is poor, so the lichen body is thin.
・ The lower skin layer is well developed. A clear border is recognized.
I wrote this in the drawing.

(5) Comparison of lichen components When the lichen components of newly formed small pieces were compared with natural ones, atranoline and decanolic acid were detected. This is the same component as that originally contained in the natural lichen body, and is evidence that the proliferation of the lichen body by redifferentiation was confirmed.

"Summary of invention"
The following is a summary based on one embodiment of the present invention.

A 7mm square lichen body piece 1 cut out from the periphery of the lichen body of Umenokigoke was fixed to the cedar bark 2 with nylon mesh 3, and cultured for 15 months. Redifferentiation of the lichen body 100 (debris) was confirmed. That is, it is considered that a bud-like thing is formed from a part including a part of the algal layer or the medulla layer, and the formation of this bud starts and regeneration begins. And the process of this fragment formation is guessed as follows. That is, in the initial stage of formation, small protrusions 4 are formed from the cortex or medulla layer containing algae. They are very similar in form to the idea formed from the epithelial layer. Further, the small protrusions extend into a cylindrical shape, and eventually, they coalesce with each other to form a palm-shaped piece. Umenochoke contains reganolic acid and atranoline as secondary metabolites, but the same two components were also detected in the newly formed fragment.

"Example of use"
As other use examples of the present invention, as shown in FIGS. 14 to 16, there is an example used for ornamental purposes. Is also possible.

Moreover, although not shown in figure, there exists an example utilized as a biological parameter | index. In this example, a small genus mushroom fragment is attached to a suitable board, and it is hung on a tree branch such as a cherry tree in the school. After being left for several months, the state of air pollution can be known from the state of the genus mushroom. Therefore, it can be used as an effective material for environmental education for elementary and junior high school children and students.

This is an ecological photograph of the lichen used in the experiment, showing the figure at the start of transplantation on May 5, 2001 This is an ecological photograph of the lichen used in the experiment, showing a figure about 4 months after the start of transplantation on September 27, 2001. This is an ecological photograph of the lichen used in the experiment, showing a figure about 15 months (1 year and 3 months) after the start of transplantation on August 3, 2002 The figure which showed the surface of the external form of a lichen The figure which showed the back of the external form of a lichen A diagram showing the surface of a differentiated lichen body, taken with a scanning electron microscope. Figure showing the back side of a differentiated lichen body, taken with a scanning electron microscope. The image of the lichen body development stage taken with a scanning electron microscope, showing the newly formed sprouting on the surface of the transplanted lichen body A photograph showing the stage of lichen formation with a scanning electron microscope, showing an extremely early lichen body on the cut surface, and a state in which the external form resembles fissure A figure showing the lichen body in a slightly grown state, taken with a scanning electron microscope. A figure showing a lichen body that has been fully developed in the shape of a palm, taken with a scanning electron microscope. The figure shows the initial state of the lichen body taken with a microscope. Figure showing the state of the lichen body taken with a microscope and showing the state of the late stage Front view of the lichen body of the present invention planted on a large bonsai set up in the lobby of a hotel, inn, etc. Front view of the lichen body of the present invention planted on a small bonsai mainly for home use Front view of the lichen body of the present invention planted on a tree or the like in a garden

Explanation of symbols

1 Lichen body piece
100 lichen
2 Cedar bark
3 Nylon mesh
4 Small protrusion
H Tree
H1 Bonsai

Claims (4)

(1) Cut out a piece of lichen from the surrounding area of a large leaf-like lichen growing in the field,
(2) The cut lichen body is transplanted to the surface of a tree species such as cedar, plum, pine, or the base material such as stone,
(3) The transplanted lichen body is covered with a mesh material such as nylon mesh or vinyl mesh for preventing the dropout, and the lichen body is fastened to the base with fasteners such as staples, nails, pins, adhesives,
(4) By the formation of a hemispherical protrusion from the algal layer in the fastened lichen body, and by the expansion of this protrusion, an ideal shape expansion, and by this expansion, by fusion of the adjacent small fragments of the ideal shape , The developmental stage of expanding the isidea-shaped fragment into a palm shape;
(5) A leaf shape characterized in that it fixes to the substrate and proliferates the lichen body through the hypodermis false roots that occur at the time when the expanded small fragments (broken-like ones) are formed. Artificial breeding method for lichens outdoors. (1) Cut out a square lichen body with a length of 4mm to 10mm and a weight of 2mg to 8mg from the periphery of a large leaf-like lichen growing in the field,
(2) Transplant the cut-out square lichen onto the surface of a tree species such as cedar, plum, pine, or a base such as stone,
(3) The transplanted lichen body is covered with a mesh material such as nylon mesh or vinyl mesh for preventing the dropout, and the lichen body is fastened to the base with fasteners such as staples, nails, pins, adhesives,
(4) Formation of hemispherical protrusions from the algae layer in the clamped lichen body and the expansion of the protrusions by the growth of the protrusions. A developmental stage in which the icidal fragment is expanded into a palm by fusion;
(5) At the stage where this medullary layer is covered with the lower skin layer, and the medullary layer is covered by the lower skin layer through the inferior hypodermis false root that occurs at the time when this expanded small fragment (split-like one) is formed. After this hypodermis layer is formed with the mycelium of the medulla layer,
(6) A method for artificially proliferating a leaf-shaped lichen body in the field, characterized in that the development of the lower skin layer or the proliferation of the lichen body is intended. The foliate lichen produced by the artificial propagation method in the field according to claim 1 or claim 2 is used for the foliage lichen grown by the artificial propagation method, characterized by being vegetated on bonsai soil or trees. object. A leaf-like lichen produced by the artificial propagation method in the field according to claim 1 or claim 2 is used as a biological indicator of air pollution, and a leaf-like lichen grown by the artificial growth method is used. object.

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