Fasteners and fastener making kits

Abstract

To improve a survival rate of grafts.SOLUTION: A fastener (100) is a fastener to fasten a plant to be an object of grafting. The fastener (100) is gelatinous.SELECTED DRAWING: Figure 1

Description

[Technical field]

[0001] The present invention relates to fasteners and fastener making kits. [Background technology]

[0002] Grafting is a method of growing a new plant by grafting a branch or bud of a plant onto another plant. Grafting can prevent problems caused by continuous cropping, improve quality and yield, and propagate new varieties.

[0003] Traditionally, grafting has involved the grafting of relatively mature plants, but in recent years, research has been conducted into micrografting, which involves grafting of young plantlets (Non-Patent Document 1).Non-Patent Document 1 describes how using the apical part of an eggplant shoot as a scion in a test tube inhibits re-callus formation in eggplant adventitious buds and promotes growth. [Prior art documents] [Non-patent literature]

[0004] [Non-Patent Document 1] Yuji Yamamoto, Satoshi Matsumoto, "Inhibition of re-callus formation and promotion of growth of eggplant adventitious shoots by micrografting", Journal of the Japanese Society of Horticulture, ISSN00137626, Vol. 63, No. 1, June 1994, pp. 67-72 Summary of the Invention [Problem to be solved by the invention]

[0005] Generally, grafting requires a certain degree of practice, and the plant to be grafted may not take root properly. In particular, in the micrografting method described in Non-Patent Document 1, the plants to be handled are very small, so the rate of grafting is likely to be low.

[0006] The present invention has been made in consideration of the above problems, and its object is to provide a fastener and a fastener manufacturing kit that can improve the survival rate of grafts. [Means for solving the problem]

[0007] According to one aspect of the present invention, the fastener is a fastener for fastening a plant to be grafted. The fastener is in a gel form.

[0008] According to another aspect of the present invention, a fastener making kit is a fastener making kit for making a fastener for fastening a plant to be grafted. The fastener making kit includes a container, a gel-like material containing agar contained in the container, and a separating tool that is fitted into the container and separates the gel-like material into a plurality of blocks. The separating tool has a lattice portion, a support plate that supports the lattice portion, and a protrusion provided on the support plate.

[0009] According to another aspect of the present invention, a fastener making kit is a fastener making kit for fastening a plant to be grafted. The fastener making kit includes a powder for preparing a gel-like material containing agar, a container capable of containing the gel-like material, and a separating tool that is fitted into the container and separates the gel-like material into a plurality of blocks. The separating tool has a lattice portion, a support plate that supports the lattice portion, and a protrusion provided on the support plate. Effect of the Invention

[0010] According to the present invention, the survival rate of grafts can be improved. [Brief description of the drawings]

[0011] [Figure 1] 1(a) and 1(b) are schematic perspective views of the fastener of the present embodiment. [Diagram 2] 1(a) to 1(d) are schematic perspective views showing grafting of a plant using the fastener of the present embodiment. [Diagram 3] 1(a) to 1(g) are schematic diagrams illustrating a method for producing the fastener of the present embodiment. [Figure 4] 1(a) to 1(c) are schematic diagrams illustrating a method for producing the fastener of the present embodiment. [Diagram 5]1(a) to 1(d) are schematic diagrams illustrating a method for producing the fastener of the present embodiment. [Figure 6] 1(a) to 1(c) are schematic diagrams illustrating a method for producing the fastener of the present embodiment. [Figure 7] 1(a) to 1(c) are schematic diagrams of a fastener producing kit for producing the fastener of the present embodiment. [Figure 8] 1 is a graph showing the graft survival rate in the treatment area where the fastener of Example 1 was attached. [Figure 9] 1 is a graph showing the graft survival rate in the treatment areas where the fasteners of Examples 1 and 2 were attached. [Figure 10] 1 is a graph showing the graft survival rate in the treatment areas where the fasteners of Examples 1 and 3 were attached. [Figure 11] 1 is a graph showing the graft survival rate in the treatment areas where the fasteners of Examples 1 and 4 were attached. [Figure 12] 1 is a graph showing the graft survival rate in the treatment areas where the fasteners of Examples 1 and 5 were attached. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0012] Hereinafter, an embodiment of a fastener and a fastener making kit according to the present invention will be described with reference to the drawings. In the drawings, the same or corresponding parts are denoted by the same reference numerals and will not be described repeatedly. In this specification, in order to facilitate understanding of the invention, the orientation of the fastener may be described with reference to the mutually orthogonal X-axis, Y-axis, and Z-axis. For example, the Z-axis direction indicates the vertical direction, and the X-axis and Y-axis directions indicate the horizontal direction. In addition, in this specification, in order to facilitate understanding of the invention, the orientation of the gel-like material may be described with mutually orthogonal x-axis, y-axis, and z-axis. For example, the z-axis direction indicates the vertical direction, and the x-axis and y-axis directions indicate the horizontal direction.

[0013] First, a fastener 100 of this embodiment will be described with reference to Fig. 1. Fig. 1(a) and Fig. 1(b) are schematic perspective views of the fastener 100. The fastener 100 is used for grafting. The fastener 100 can suitably join multiple plants together.

[0014] As shown in Fig. 1(a), the fastener 100 is solid. The fastener 100 fastens two plants in a joined state. The fastener 100 can suitably join one plant to another plant. For example, the fastener 100 can join two plants by fastening the cross sections of the scion and the rootstock in an adhesive state.

[0015] <Fastener> The fastener 100 is in a gel state. Typically, the fastener 100 is also called a jelly-like material. The fastener 100 is in a gel state.

[0016] Fastener 100 includes a gelling substance for causing fastener 100 to become a gel. Typically, the gelling substance gels a liquid. Typically, the liquid to be gelled is an aqueous solution. Fastener 100 has the gelling substance dispersed substantially uniformly therein.

[0017] Fastener 100 may contain agar. In fastener 100, agar contains agarose. For example, in fastener 100, the mass concentration of agarose is 0.5% or more and 2.0% or less. Typically, the higher the agarose concentration, the more polysaccharide components there are, but fastener 100 tends to become hard. Conversely, the lower the agarose concentration, the softer fastener 100 becomes, making fastener 100 more difficult to handle.

[0018] The fastener 100 may further include chitin. Examples of chitin include chitin, chitin nanofiber, and chitosan.

[0019] In the fastener 100, the concentration of chitin and chitin nanofibers may be 0.0001% or more and 1.0% or less, or 0.025% or more and 0.2% or less, which can improve the survival rate of the graft.

[0020] Additionally, the fastener 100 may further include at least one of an auxin and a cytokinin.

[0021] For example, in fastener 100, the gelling substance is agar. Alternatively, the gelling substance may be gellan gum or gelatin.

[0022] The fastener 100 has a main body portion 110 and a holding portion 120. Typically, the main body portion 110 has a substantially rectangular parallelepiped shape. The main body portion 110 may be a gel-like material.

[0023] The holding portion 120 is provided on the surface of the main body portion 110. The holding portion 120 holds multiple plants to be grafted in a contact state. Here, the holding portion 120 is provided on the -Z direction side of the main body portion 110. Typically, the holding portion 120 is provided in the center of one surface of the main body portion 110. It is preferable that the holding portion 120 extends from one end to the other end on the surface on which the main body portion 110 is provided.

[0024] For example, the holding portion 120 includes a groove or ridge provided on the surface of the main body portion 110. The holding portion 120 may be formed integrally with the main body portion 110. Alternatively, the holding portion 120 may be formed by cutting the surface of the main body portion 110. The holding portion 120 holds the plants by inserting multiple plants to be grafted.

[0025] In detail, the main body 110 has a first surface 110a, a second surface 110b, a third surface 110c, a fourth surface 110d, a fifth surface 110e, and a sixth surface 110f. The first surface 110a is located on the +Z direction side, and the second surface 110b is located on the -Z direction side. The third surface 110c, the fourth surface 110d, the fifth surface 110e, and the sixth surface 110f are located on the -X direction side, the -Y direction side, the +X direction side, and the +Y direction side, respectively. Here, the first surface 110a, the second surface 110b, the third surface 110c, the fourth surface 110d, the fifth surface 110e, and the sixth surface 110f are all flat surfaces.

[0026] Here, the holder 120 is provided on the second surface 110b of the main body 110. The holder 120 extends parallel to the Y direction through the center of a line along the X direction of the second surface 110b. The holder 120 extends on the second surface 110b from an end of the fourth surface 110d to an end of the sixth surface 110f.

[0027] As shown in FIG. 1(b), the fastener 100 fastens the plants P1 and P2 in a contacting state. Here, the plants P1 and P2 have portions that extend linearly in the Y direction. The fastener 100 holds the contact portion (contact portion) Pc of the plants P1 and P2. The holding portion 120 holds the plants P1 and P2. It is preferable to hold the plants P1 and P2 by contacting the sides of the contact portion Pc. Typically, the thickness (length along the X direction) of the plants P1 and P2 is approximately equal to the width (length along the X direction) of the holding portion 120.

[0028] In this embodiment, the fastener 100 containing agar holds the contact portion Pc of the plants P1 and P2, thereby promoting connection and growth of the joint between the plants P1 and P2 while keeping the contact portion Pc moist, and enabling the plants P1 and P2 to be efficiently grafted.

[0029] The fastener 100 of this embodiment is suitable for use in micrografting. For example, the thickness (length along the X direction) of the plants P1 and P2 is 0.5 mm or more and 5 mm or less. Typically, micrografting is performed in a test tube or a petri dish. It is preferable that micrografting is performed in a sterile environment.

[0030] In this case, for example, the length of the main body 110 along the X direction may be 2 mm or more and 20 mm or less, or may be 3 mm or more and 15 mm or less. The length of the main body 110 along the Y direction may be 2 mm or more and 20 mm or less, or may be 3 mm or more and 15 mm or less. The length of the main body 110 along the Z direction may be 2 mm or more and 20 mm or less, or may be 3 mm or more and 15 mm or less. In addition, the length (width) of the holding portion 120 along the Y direction may be 0.5 mm or more and 2 mm or less.

[0031] As described above, the fastener 100 of the present embodiment is suitable for use in micrografting. For example, the fastener 100 can be used to suitably graft shoot tips or seedlings. By grafting seedlings, grafting can be performed in a small space and the survival rate can be improved.

[0032] Next, grafting of plants P1 and P2 using the fastener 100 of this embodiment will be described with reference to Figures 1 and 2. Figures 2(a) to 2(d) are schematic diagrams for explaining grafting of plants P1 and P2 using the fastener 100.

[0033] As shown in Fig. 2(a), a fastener 100 is prepared. The fastener 100 has a main body portion 110 and a holding portion 120. Typically, the main body portion 110 has a substantially rectangular parallelepiped shape. The holding portion 120 is provided on the outer surface of the main body portion 110. The holding portion 120 is provided in the center of one surface of the main body portion 110.

[0034] 2(b), plants P1 and P2 are prepared. Plants P1 and P2 are arranged so that an end of plant P1 and an end of plant P2 are in contact with each other at contact portion Pc.

[0035] For example, the plant P1 and the plant P2 are arranged on the culture medium Cm so as to be in contact with each other. The culture medium Cm may contain agar. The culture medium Cm containing agar as a main component is also called an agar medium. The culture medium Cm may be Murashige and Skoog medium (MS medium). When the culture medium Cm contains agar, the components of the culture medium Cm may be the same as or different from the components of the fastener 100.

[0036] Furthermore, when the medium Cm contains agar, the agarose concentration of the medium Cm may be higher than the agarose concentration of the fastener 100. However, the agarose concentration of the medium Cm may be approximately the same as the agarose concentration of the fastener 100, or may be lower than the agarose concentration of the fastener 100. Alternatively, the medium Cm may contain gellan gum.

[0037] As shown in FIG. 2(c), the fastener 100 is placed on either the plant P1 or the plant P2. Here, the fastener 100 is placed on the plant P2. Therefore, the holding portion 120 of the fastener 100 holds the plant P2. For example, the plant P2 is inserted into the holding portion 120 of the fastener 100. At this time, the fastener 100 does not cover the contact portion Pc between the plant P1 and the plant P2. By not placing the fastener 100 directly on the contact portion Pc between the plant P1 and the plant P2, it is possible to prevent the plant P1 and the plant P2 from being displaced due to an impact.

[0038] As shown in FIG. 2(d), the fastener 100 is moved relative to the plants P1 and P2. Here, the fastener 100 moves on the medium Cm along the Y direction in which the plants P1 and P2 extend. As the fastener 100 moves, the fastener 100 covers the contact portion Pc between the plants P1 and P2. This allows the fastener 100 to fasten the plants P1 and P2 while holding them. When micrografting is performed, the fastener 100 may be moved while the plants P1, P2, and the fastener 100 are observed under a microscope.

[0039] Thereafter, the fastener 100 is left in a state in which it holds the plants P1 and P2. Typically, the fastener 100 is left in a state in which it holds the plants P1 and P2 for several days. This allows the plants P1 and P2 to be grafted.

[0040] Before the fastener 100 covers the contact points between the plants P1 and P2, the plants P1 and P2 are arranged to extend horizontally, and the fastener 100 covers the plants P1 and P2 from above vertically (+Z direction), but this embodiment is not limited to this. The fastener 100 may cover the plants P1 and P2 horizontally. However, when the fastener 100 is attached so as to cover the contact points between the plants P1 and P2, it is preferable that the fastener 100 covers the plants P1 and P2 from above vertically (+Z direction). In this case, after the fastener 100 holds the plants P1 and P2, the fastener 100, the plants P1, and the plants P2 may be moved together in any direction.

[0041] According to this embodiment, the plants P1 and P2 are fastened by the fastener 100 containing agar, so that the survival rate of the plants P1 and P2 can be improved.

[0042] Next, a method for producing the fastener 100 of this embodiment will be described with reference to Figs. 3(a) to 3(g).

[0043] As shown in Fig. 3(a), a gel-like material Ge containing agar is formed. In the gel-like material Ge, the agar contains agarose. The gel-like material Ge may further contain chitin.

[0044] Here, the gel-like material Ge has a substantially rectangular parallelepiped shape. In one example, the gel-like material Ge in FIG. 3(a) is prepared in the same manner as a so-called typical agar medium. For example, the gel-like material Ge may be formed in a square petri dish.

[0045] As shown in FIG. 3(b), a plurality of incisions Ca extending parallel to each other are formed in the gel-like material Ge. The incisions Ca are formed with tweezers or a knife. For example, the incisions Ca are linearly cut into the gel-like material Ge along the x direction. The depth of the incisions Ca is the thickness of the gel-like material Ge (length in the z direction). For example, the interval between the plurality of incisions Ca (distance between them in the y direction) corresponds to the width of the fastener 100. The interval between the plurality of incisions Ca is 3 mm or more and 10 mm or less.

[0046] As shown in FIG. 3(c), a plurality of incisions Cb are formed in the gel-like material Ge, extending parallel to each other in a direction different from the direction in which the incisions Ca extend. The incisions Cb are formed with tweezers or a knife. Here, the plurality of incisions Cb are formed along a direction perpendicular to the previously incised incisions Ca. For example, the incisions Cb are linearly incised in the gel-like material Ge along the y direction. The depth of the incisions Cb is the thickness (length in the z direction) of the gel-like material Ge. For example, the interval between the plurality of incisions Ca (the distance between them in the y direction) corresponds to the length of the fastener 100. The interval between the plurality of incisions Cb (the distance between them in the x direction) is 3 mm or more and 15 mm or less. For example, the interval between the incisions Cb may be greater than the interval between the incisions Ca.

[0047] A gel block Gb is formed by forming a plurality of cuts in two different directions in the gel material Ge.

[0048] 3(d), the block Gb is taken out from the gel-like material Ge. For example, the block Gb is taken out from the gel-like material Ge using tweezers. Here, the block Gb has a substantially rectangular parallelepiped shape.

[0049] As shown in FIG. 3(e), a groove Cg is formed in the block Gb. The groove Cg is formed with tweezers or a knife. The groove Cg is formed relatively shallowly in the center of one main surface of the block Gb from one end to the other end. For example, the groove Cg is formed so as to divide the long side of the block Gb into two equal parts. The depth of the groove Cg is, for example, 0.5 mm or more and 2 mm or less.

[0050] The block Gb having the groove Cg formed therein is preferably used as the fastener 100. Here, the groove Cg of the block Gb functions as the holding portion 120 of the fastener 100.

[0051] 3(f), the fastener 100 is placed on one of the plant P1 and the plant P2. Here, the fastener 100 is placed on the plant P2. The holding portion 120 of the fastener 100 covers the plant P2.

[0052] 3(g), the fastener 100 is slid relative to the plants P1 and P2, so that the fastener 100 holds the contact portion Pc of the plants P1 and P2.

[0053] In this manner, the fastener 100 can be produced. According to this embodiment, the fastener 100 can be produced by partially cutting out and processing the gel material Ge.

[0054] Next, another method for producing the fastener 100 of this embodiment will be described with reference to Figures 4 to 6. Figure 4 shows chemicals and parts prepared when producing the fastener 100 of this embodiment.

[0055] As shown in Figs. 4(a) to 4(c), in order to prepare the fastener 100, mixed salts for MS medium Mp1, vitamin powder for MS medium Mp2, agar powder Mp3, container Dc, container Mc and separation tool Se are prepared.

[0056] 4(a) is a schematic diagram of mixed salts Mp1 for MS medium, vitamin powder Mp2 for MS medium, container Dc, and container Mc. Mixed salts Mp1 for MS medium is sealed in a bag. Mixed salts Mp1 for MS medium may contain any of KNO3, NH4NO3, CaCl2·2H2O, MgSO4·7H2O, KH2PO4, Na2-EDTA, FeSO4·7H2O, MnSO4·4H2O, ZnSO4·7H2O, H3BO3, KI, Na2MoO4·2H2O, CuSO4·5H2O, and CoCl2·6H2O.

[0057] The vitamin powder Mp2 for MS medium may contain any one of glycine, myoinositol, nicotinic acid, pyridoxine hydrochloride, and thiamine hydrochloride. The vitamin powder Mp2 for MS medium is sealed in a bag. Here, the vitamin for MS medium is in powder form, but the vitamin for MS medium may be in liquid form.

[0058] Agar powder Mp3 is a dry agar in powder form.

[0059] The container Dc is used to dissolve the mixed salts Mp1 for MS medium, the vitamin powder Mp2 for MS medium, and the agar powder Mp3 in a liquid. Typically, water is added to the container Dc together with the mixed salts Mp1 for MS medium, the vitamin powder Mp2 for MS medium, and the agar powder Mp3. Typically, in the container Dc, the mixed salts Mp1 for MS medium and the vitamin powder Mp2 for MS medium are dissolved in water. The container Dc is heated together with the dissolving liquid.

[0060] For example, the container Dc is a heat-resistant flask. In one example, the container Dc has a heat resistance of 120° C. or more.

[0061] The container Mc is used to gelatinize the solution in the container Dc to form a gel-like substance. The container Mc has a hollow box Ma. The solution is poured into the container Mc and gelled.

[0062] For example, the width of the box Ma may be 5 cm or more and 50 cm or less, or 10 cm or more and 30 cm or less. The length of the box Ma may be 5 cm or more and 50 cm or less, or 10 cm or more and 30 cm or less. Furthermore, the height of the box Ma may be 3 cm or more and 10 cm or less.

[0063] The container Mc preferably further has a lid Mb that covers the opening of the box Ma. The lid Mb can shield the dissolving solution or gel-like substance in the box Ma from the surroundings.

[0064] Next, the separation tool Se will be described with reference to Fig. 4(b) and Fig. 4(c). Fig. 4(b) is a schematic perspective view of the separation tool Se, and Fig. 4(c) is a schematic view of the separation tool Se. The separation tool Se separates the gel-like material into a plurality of blocks. The separation tool Se is heat-resistant. For example, the separation tool Se is heat-resistant to 60°C or more.

[0065] As shown in Figures 4(b) and 4(c), the separator Se has a lattice-shaped lattice portion S1. The lattice portion S1 includes a plurality of linear portions extending in a first direction and a plurality of linear portions extending in a second direction perpendicular to the first direction.

[0066] The separation tool Se is inserted into the box Ma of the container Mc. The outer diameter of the separation tool Se may be approximately equal to the inner diameter of the box Ma. This allows the separation tool Se to be fitted into the box Ma.

[0067] When the dissolving solution is gelled in the box Ma of the container Mc, the separation tool Se is fitted in the box Ma, so that the gelled material can be separated into a plurality of blocks. The separation tool Se may be fitted in the box Ma before the dissolving solution is gelled in the box Ma. Alternatively, the separation tool Se may be fitted in the box Ma after the dissolving solution is gelled in the box Ma.

[0068] 4(b), the separation tool Se may further include a handle portion S2 disposed on one side surface of the lattice portion S1. The handle portion S2 allows a user to easily handle the separation tool Se.

[0069] As shown in Figures 4(b) and 4(c), the separation tool Se may further include a support plate S3. The support plate S3 supports the lattice portion S1. The support plate S3 supports the lattice portion S1 from below.

[0070] Furthermore, as shown in FIG. 4(c), the separator Se may further have a protrusion S4 provided on the support plate S3 in addition to the lattice portion S1, the handle portion S2, and the support plate S3. The protrusion S4 protrudes in a stripe shape from the support plate S3. The height of the protrusion S4 is lower than the height of the linear portion of the lattice portion S1. Here, the protrusion S4 extends in the first direction together with a plurality of linear portions extending in the first direction and is located between the linear portions of the order.

[0071] Next, a method for producing the fastener 100 using the MS medium mixed salts Mp1, the MS medium vitamin powder Mp2, the agar powder Mp3, the container Dc, the container Mc and the separation tool Se will be described with reference to Figs.

[0072] As shown in FIG. 5(a), mixed salts for MS medium Mp1, vitamin powder for MS medium Mp2, and agar powder Mp3 are poured into a container Dc and mixed with water to prepare a solution.

[0073] As shown in Fig. 5(b), the container Dc is heated. For example, the container Dc is heated to a temperature of 60°C or higher. The agar powder Mp3 contained in the dissolving solution Ld in the container Dc is dissolved by heating. When the dissolving solution Ld in the container Dc is to be sterilized, the container Dc is preferably heated in an autoclave.

[0074] As shown in FIG. 5(c), the dissolving liquid Ld in the container Dc is poured into the container Mc.

[0075] As shown in FIG. 5(d), before the dissolving liquid Ld in the container Mc gels, the separation tool Se is placed in the container Mc.

[0076] As shown in FIG. 6(a), the box Ma is covered with a lid Mb and left at room temperature to cool the dissolving solution in the container Mc.

[0077] As shown in Fig. 6(b), the solution Ld in the container Mc is cooled, and the solution Ld gels and changes into a gel-like material Ge. Here, since a separator Se is disposed in the container Mc, the gel-like material Ge is separated into a plurality of gel-like blocks Gb. In addition, since the separator Se has a protrusion S4, grooves are formed in the plurality of separated blocks of the gel-like material Ge.

[0078] As shown in Fig. 6(c), the gel block Gb is taken out from the container Mc. The separation tool Se has a protrusion S4 on the support plate S3, so that a groove Cg is provided in the gel block Gb. Therefore, the gel block Gb can be used as a fastener 100.

[0079] According to this embodiment, the dissolving liquid Ld is gelled in a state where the separator Se having the lattice portion S1 and the protrusions S4 is disposed, so that the gelled material Ge is separated into a plurality of gelled blocks Gb during gelling and the grooves Cg are formed. Therefore, the gelled blocks Gb having the grooves Cg formed therein can be used as the fastener 100.

[0080] Next, a fastener manufacturing kit 200 of the present embodiment will be described with reference to Fig. 7. Fig. 7(a) to Fig. 7(c) are schematic diagrams of the fastener manufacturing kit 200 for manufacturing the fastener 100 of the present embodiment.

[0081] As shown in Fig. 7(a), the fastener fabrication kit 200 may be an agar gel-like material Ge. The gel-like material Ge can be cut into a block Gb of a specific size and a groove Cg cut into the block Gb to be used as the fastener 100. In this case, the fastener fabrication kit 200 is preferably stored in a sterile environment.

[0082] The agar gel Ge may be formed not only by cutting it out and cutting the grooves Cg into it, but also by dissolving it again and molding it using the container Mc and separation tool Se described above with reference to FIG.

[0083] As shown in FIG. 7(b), the fastener fabrication kit 200 may include a container Mc containing an agar gel-like material Ge and a separator Se. The container Mc is fitted with a separator Se that separates the gel-like material Ge into blocks Gb of a predetermined size and forms grooves Cg in the blocks Gb. This allows a predetermined fastener 100 to be easily removed from the agar gel-like material Ge of the fastener fabrication kit 200. In this case, the fastener fabrication kit 200 is preferably stored in a sterile environment.

[0084] Alternatively, as shown in Fig. 7(c), the fastener preparation kit 200 may include a mixed salt for MS medium Mp1, a vitamin powder for MS medium Mp2, agar powder Mp3, a container Mc, and a separation tool Se. The fastener preparation kit 200 can suitably prepare the fastener 100. The fastener preparation kit 200 may further include a container Dc shown in Fig. 4(a).

[0085] The fastener 100 can be suitably produced using the fastener production kit 200 of this embodiment. EXAMPLES

[0086] Example 1: Agar-containing fastener A dissolving solution was prepared by diluting the dissolving solution for making a normal MS medium by two times using mixed salts for MS medium (manufactured by Fujifilm Wako Pure Chemical Industries, Ltd.) and vitamin solution for MS medium (manufactured by Sigma-Aldrich). In detail, 1500 mL of distilled water was added to a 2000 mL plastic bottle, followed by the mixed salts for MS medium (one bag for 1 L). Then, 1 mL of vitamin solution for MS medium was added to the plastic bottle and stirred with a stirrer. Then, 1 mM potassium hydroxide solution was added to adjust the pH of the dissolving solution to 5.8. Then, distilled water was added to the plastic bottle to adjust the dissolving solution to 2000 mL, and the dissolving solution was prepared by stirring with a stirrer. Table 1 shows the components of the dissolving solution.

[0087] [Table 1]

[0088] Thereafter, 20 g of agar powder (manufactured by Nacalai Tesque, Inc.) was added to 2000 mL of the dissolution solution to adjust the agarose concentration to 1%.

[0089] The plastic bottle was heated in an autoclave for 2 hours. The autoclave was heated at 121°C for the first 15 minutes, and then kept at 60°C. The solution was then poured into a petri dish in a clean bench. The solution was left at room temperature to gel in the petri dish.

[0090] Thereafter, a block measuring 5 mm in length, 7.5 mm in width and 5 mm in height was cut out from the gel-like material, and a groove measuring 2.5 mm in depth and 5 mm in length was cut into one surface of this block to produce the fastener of Example 1.

[0091] [Engraftment test] Lettuce seedlings 4 to 5 days after sowing were Y-grafted and fitted with the fasteners of Example 1. After 7 days of curing, the success rate (graft survival rate) was calculated, with the case where the scion did not fall off being considered successful and the case where the scion fell off being considered unsuccessful.

[0092] Fig. 8 shows the graft survival rate when the fastener of Example 1 was used, as well as the graft survival rate when no fastener was attached as a comparative example. As shown in Fig. 8, the graft survival rate in the control area of ​​the comparative example was 45.7% (number of samples: 129), while the graft survival rate in the treatment area using the fastener of Example 1 was 68.8% (number of samples: 125), showing a significant difference (Fisher's exact test (hereinafter, unless otherwise specified, Holm's method), p = 0.0002346).

[0093] [Example 2: Chitin-containing agar fastener] Chitin was added at different concentrations and suspended in a dissolving solution having the composition shown in Table 1. The fastener of Example 2 was produced in the same manner as in Example 1. Here, the chitin concentration was adjusted to 0.3, 0.1, 0.05, 0.01, 0.001, and 0.0001 to produce the fasteners of Examples 2a to 2e.

[0094] Table 2 shows the fasteners, chitin concentrations, sample numbers, and graft survival rates for Comparative Example and Examples 1 and 2a to 2e. Figure 9 shows the results of the graft survival rates when the fasteners of Examples 1 and 2a to 2e were used, as well as the results for the graft survival rate when no fasteners were used as a Comparative Example.

[0095] [Table 2]

[0096] As shown in Figure 9, the graft survival rate was 45.7% (n = 129) in the control area of ​​the comparative example, and 68.8% (n = 125) in the treatment area where the fastener of Example 1, which does not contain chitin, was attached. In contrast, the graft survival rate increased (76.9-85.5%) in all of the control areas where the fasteners of Examples 2a-2e, which contain chitin, were attached. As shown in Table 2, the graft survival rate was significantly improved in all of the treatment areas where the fasteners of Examples 2a-2e were attached, compared to the control area of ​​the comparative example.

[0097] [Example 3: Chitin nanofiber-containing agar fastener] Chitin nanofibers were added at different concentrations and suspended in a dissolving solution having the composition shown in Table 1 above. The fastener of Example 3 was produced in the same manner as in Example 1. Here, the chitin concentration was changed to 0.3, 0.2, 0.15, 0.1, 0.05, 0.025, 0.01, 0.001, and 0.0001 to produce the fasteners of Examples 3a to 3i.

[0098] Table 3 shows the fasteners, chitin nanofiber concentrations, sample numbers, and graft survival rates for Comparative Example, Example 1, and 3a to 3i. Figure 10 shows the results of the graft survival rates when the fasteners of Example 1 and 3a to 3i were used, as well as the graft survival rate when no fastener was used as a Comparative Example.

[0099] [Table 3]

[0100] As shown in FIG. 10, the graft survival rate was 45.7% in the control area without the fastener of the comparative example, and 68.8% in the treatment area with the fastener of Example 1 (agar fastener not containing chitin nanofiber), while the graft survival rate increased in all areas with the fastener of Example 3 (78.6% to 93.75%). The graft survival rate of all of the treatment areas of Example 3 was significantly improved compared to the control area. On the other hand, when compared with the treatment area with the fastener of Example 1 (agar fastener not containing chitin nanofiber), the graft survival rate was significantly improved for the treatment area with the fastener of Example 3b (fastener containing chitin nanofiber at a concentration of 0.2%) (p=0.014) and the treatment area with the fastener of Example 3d (fastener containing chitin nanofiber at a concentration of 0.1%) (p=0.0078).

[0101] [Example 4] [Auxin / cytokinin-containing agar fastener] Auxin (Oxyberon, manufactured by Bayer CropScience) or cytokinin (Fulmet, manufactured by Sumitomo Chemical Co., Ltd.) was added at different concentrations to the solution having the composition shown in Table 1 above and suspended in the solution. The rest was the same as in Example 1, and the fasteners of Example 4 were produced. Here, the (auxin, cytokinin) was adjusted to (2 mg / L, 0 mg / L), (2 mg / L, 0.2 mg / L), (2 mg / L, 1 mg / L), (4 mg / L, 0 mg / L) and (2 mg / L, 0.4 mg / L) to produce the fasteners of Example 4a to Example 4e.

[0102] Table 4 shows the clamps, auxin or cytokinin concentrations, sample numbers, and graft survival rates for Comparative Example, Example 1, and 4a to 4e. Fig. 11 shows the results of the graft survival rates when the clamps of Example 1 and 4a to 4e were used, as well as the graft survival rate when no clamp was used as a Comparative Example.

[0103] [Table 4]

[0104] As shown in Figure 11, the graft survival rate was 45.7% in the control area without the clamp of the comparative example, and 68.8% in the area with the agar clamp of Example 1, whereas the graft survival rate was significantly improved to over 90% in the treatment areas with the clamps of Examples 4b, 4c and 4e containing both auxin and cytokinin. On the other hand, in the treatment areas with the clamps of Examples 4a and 4d containing auxin but not cytokinin, the graft survival rate was 50% for the clamp of Example 4a and 64.8% for the clamp of Example 4d, which was lower than the area with the clamp of Example 1.

[0105] [Example 5: Agarose concentration] Agarose was added at different concentrations to the dissolving solution having the composition shown in Table 1 above and suspended. The fastener of Example 5 was prepared in the same manner as in Example 1 except for the above. Here, the agarose concentrations were adjusted to 0.4%, 0.5%, 0.6%, 0.8%, 1.2%, 1.4%, 1.6% and 1.8% to prepare the fasteners of Examples 5a to 5h. As mentioned above, in the fastener of Example 1, the agarose concentration was adjusted to 1%.

[0106] Table 5 shows the agarose concentration of the fasteners of Comparative Example, Example 1, and 5a to 5h, the number of successful grafts, the number of failed grafts, the total number of tests, and the success rate. Figure 12 shows the success rate (graft survival rate) when the fasteners of Example 1 and 5a to 5h were used, as well as the success rate when no fastener was attached as a Comparative Example.

[0107] [Table 5]

[0108] As shown in Table 5, the success rate (grafting survival rate) of the treatments with the fasteners of Examples 1 and 5a to 5e (agarose concentrations 1.0%, 0.4%, 0.5%, 0.6%, 0.8%, 1.2%, 1.4%, 1.6% and 1.8%) was higher than that of the treatments without the fasteners of the comparative example. In detail, except for the fasteners of Examples 5a and 5c (agarose concentrations 0.4% and 0.6%), the success rate (grafting survival rate) of the treatments with the fasteners of Example 1 (agarose concentration 1.0%) was the highest, and the success rate of the corresponding treatments decreased as the agarose concentration moved away from 1.0%.

[0109] The embodiments of the present invention have been described above with reference to the drawings. However, the present invention is not limited to the above embodiments, and can be implemented in various aspects without departing from the gist of the present invention. In addition, various inventions can be formed by appropriately combining multiple components disclosed in the above embodiments. For example, some components may be deleted from all components shown in the embodiments. Furthermore, components across different embodiments may be appropriately combined. The drawings are mainly shown schematically for ease of understanding, and the thickness, length, number, spacing, etc. of each component shown in the drawings may differ from the actual ones due to the convenience of drawing. In addition, the material, shape, dimensions, etc. of each component shown in the above embodiments are examples and are not particularly limited, and various changes are possible within a range that does not substantially deviate from the effects of the present invention. [Industrial Applicability]

[0110] The present invention is suitably used for fasteners and fastener manufacturing kits. [Explanation of symbols]

[0111] 100 Fasteners 110 Main body 120 Holding part 200 Fastener Making Kit Mc container Ge gel Se separation tool

Claims

1. A fastener making kit for making a fastener for fastening a plant to be grafted, A container; A gel-like material contained in the container; a separation tool that is fitted into the container and separates the gel-like material into a plurality of blocks; Equipped with The separation tool is A lattice portion; A support plate that supports the lattice portion; A protrusion portion provided on the support plate; A fastener making kit comprising:

2. A fastener making kit for making a fastener for fastening a plant to be grafted, A powder for preparing a gel-like material; A container capable of containing the gel-like material; a separation tool that is fitted into the container and separates the gel-like material into a plurality of blocks; Equipped with The separation tool is A lattice portion; A support plate that supports the lattice portion; A protrusion portion provided on the support plate; A fastener making kit comprising:

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