JPH0326495A - Preparing method for culture section of plant organization - Google Patents

Abstract

PURPOSE:To prepare a culture section of a plant organization capable of culturing without the necrosis of a cut section even being used for a tissue culture easily and in a large quantity by cutting in a slice a plant body subjected to sterile or sterilizing treatment under sterile conditions with its feeding on a blade driven by rotation. CONSTITUTION:A blade 5 is fitted onto a disk 3 rotated and driven by a motor 2 so that a blade tip 5a is faced to the external part from the internal part in the diameter direction and a gap exists properly with the disk 3. A plant body, the organ or tissue of a plant subjected to a sterile or sterilizing treatment under sterile conditions is fed from a feeding port 6 onto this blade 5 driven by rotation and cut in a slice-like. A culture section used for a tissue culture can thus be produced in a mass quantity.

Description

DETAILED DESCRIPTION OF THE INVENTION FIELD OF APPLICATION OF INDUSTRY-E The present invention relates to a method for preparing a cultured section of a plant tissue for obtaining a large scale.

BACKGROUND OF THE INVENTION Conventionally, sections of plant organs, tissues, etc., which are used in plant tissue culture and serve as culture materials, have been obtained by cutting them with a scalpel or by punching them with a cork borer. This method has the advantage of allowing the operator to determine which plant or part of the plant has been cultured the most, and to cut sections in the form most suitable for culture. The drawback was that it required a great deal of effort and time.

Problems to be Solved by the Invention In the manual method of preparing culture sections as described above,
J:. For the reasons mentioned above, it is not suitable for commercial mass production, even if the culture is carried out at an experimental level.

Mechanical cutting methods can be considered as a mass production technology, but if this method is initially adopted as the method for creating culture sections,
The cut sections were necrotic and considered undesirable. However, when we actually created cultured sections using a mechanical cutting method and cultured them, we found that if the appropriate mechanical cutting method was used depending on the type of plant, there would be no problems and there would be no hindrance to the culture. We found an unexpected result: this does not occur. The present invention was made based on this knowledge, and its purpose is to provide a technology for mass production of cultured slices for mm culture. According to the present invention, the blade is mounted on a disc rotationally driven by a motor so that the cutting edge faces outward from the inside in the radial direction, and with a suitable gap between the blade and the disc. A plant body, plant organ, or tissue piece (herein simply referred to as a plant body) that has been sterilized or sterilized under aseptic conditions is fed onto a rotationally driven blade and cut into slices. A number of methods are provided. Here, the cutting edge of the blade may be formed to pass through the center of rotation, or may be eccentric.

In any case, it is better to withdraw rather than push through.
Therefore, the cutting edge is preferably formed in an arc rather than a straight line. When pressing, the cutting becomes dull and tends to get clogged at the cutting edge, causing significant damage to the plant tissue at the cutting surface.
This is because when cutting, the sharpness is relatively good, there is less clogging at the cutting edge, and damage to the vegetable dandelion can be reduced.

The plants targeted by the present invention are of any type, but
This method is most suitable for lily bulbs, potatoes, and other bulbs.

If the plant to be fed onto the blade is soft, such as the root of a lily, you can simply place it on the blade to obtain cultured sections cut into slices, but if the plant is hard, the extent of the cutting may vary. 50-500g depending on
/One presser foot pressure is applied.

To press the plant body against the blade, for example, a predetermined weight is placed on the plant body, a compression spring whose spring pressure is adjusted to an appropriate strength is applied via a presser plate, or It is a good idea to provide a presser plate that can move forward and backward to push it in. The gap between the cutting edge of the blade and the disc and the rotational speed of the blade (disc) are preferably 1 to 4fi and 100 to 4, respectively.
The speed is set at 00 rpm, and the speed is adjusted within that range depending on the plant body and its parts that are the material for culture.

A sterile room is usually used for cutting under sterile conditions. Figures 1 and 2 show an example of the apparatus used in this method, in which a circular plate 3 rotated by a motor 2 is disposed inside a cylindrical container 1 with a bottom, and the circular plate 3 has a center of rotation. An arcuate groove 4 is bored from the vicinity toward the periphery, and a blade 5 is removably attached with a screw 8 along the groove.
The cutting edge 5a is located on the arcuate groove 4, leaving a certain gap with the upper surface of the disk. A lid 7 with a protruding supply port 6 is also removably attached to the container l so that plants are supplied through the supply port 6, and a container (not shown) is placed on top of the supplied plant depending on the type. An appropriate weight can be placed on it via a holding plate. According to this device, the plant body supplied from the supply port 6 as the disk 3 is rotated is cut into slices by the cutting edge 5a of the blade 5, falls through the arcuate groove 4, and is stored in the container. When a certain amount of water has been accumulated, the lid 7 and disk 3 are removed, and the sliced culture section is taken out. The cultured sections can also be taken out from the cylindrical container l by providing an opening. Note that the blade 5 is replaced with a blade having an appropriate thickness when it becomes dull or when it is desired to change the gap between the cutting edge 5a and the upper surface of the disk in order to change the thickness of the culture section.

The gap between the cutting edge 5a of the blade 5 and the upper surface of the disk can also be adjusted by providing a spacer of an appropriate thickness between the blade and the disk.

16 to 18 show another example, in which a disc 53 is disposed in a container 5l having a circular cross section, and is connected to a separately provided motor 52 at the top thereof by a flexible drive shaft. It is designed to be rotationally driven. As shown in FIGS. 17 and 18, long grooves 54 extending from the inner side toward the periphery are bored in the corners of the disk 53 for a certain distance, and as the blades 55 are twisted, the blades 55 are inserted through the grooves 56 and screwed into the screws 57 along the respective grooves 54. Removable ζ, 2 digits {4 digits).

This device is entirely made of metal so that it can be sterilized by heat. 1. On the side of the container, there is a reservoir for the sliced slices, L.
A viewing window 59 is provided so that the user can check the condition. 60 is the {+t candy mouth of the plant body.

In addition, 6l is a transfer station for discharging cut Sawata culture sections.
'', 62 is an air outlet r, and air is sent through a filter 63 to force the cultured slices stored in the container through the transplant D 6 1 and 7. In this device as well, a heavy L is placed on the second node of the plant supplied to the supply port, and as the node is replaced. (The gap between the cutting edge and the disk can be changed.

Another invention for preparing cultured sections includes a cutting device in which a movable blade is arranged between plow plates installed at appropriate intervals, and that rotates, moves or reciprocates between the plow plates, and a plant that has been sterilized or sterilized. A method is provided for plowing the plant body with a plow board and cutting it with a movable blade by pushing either side of the body toward another force under sterile conditions. FIG. 6 shows an example of a cutting device used in this method. Inside a box-shaped container 11, excitation plates 12 are arranged in parallel at a certain interval above and below one side of the container 11, and each plow is 6l5, which is arranged between the plates and consists of a rotary blade 14 fixed to a drive shaft 13 passing through the plow board 12 and driven to rotate, is a presser plate installed on the other side of the container l1 so as to move forward and backward. , Supply Eco16
When a suitable amount of the supplied plants accumulates in the container 23, the cutting device moves the plants forward and pushes them along with the cutting device.

The plant body cut by the cutting device is discharged from the outlet 17, but it may also be temporarily stored behind the cutting device and taken out from the corpse.

In the above cutting device, a rotary blade is arranged between the plow plates, but instead of the rotary blade, a swinging blade may be used, or a movable blade that reciprocates between the plow plates may be used. FIG. 7 shows another embodiment of the cutting device, in which a clipper-like cutting device is provided, in which fixed blades 20 are arranged in parallel at regular intervals below one in a box-shaped container l9, and an adjacent fixed blade 1 Blade 2
Slides in contact with the fixed blade between 0 (7) and reciprocates up and down G (wheeze L)
2), and a movable blade 2l that cooperates with the fixed blade 20 to cut the plant body.

22 is an ilffi retractable presser plate.

Instead of this, the movable blade of the present cutting device may be a common movable blade that slides the fixed blade along each fixed blade L2 and moves downward downward at -L.

In yet another invention for preparing culture slices, a plurality of rollers 2-41 are provided, and on the delivery side between the rollers, the axis is parallel to or perpendicular to the axis of the rollers. - A cutting position consisting of a movable blade that reciprocates in parallel with the roller, and a sterilized or sterilized plant being transferred to the other under aseptic conditions. Does “pause” mean to impose? ．． This provides a method in which the plant is sent out between the rollers and cut with a movable blade.

Figures 8 and 9 show an example of a cutting device used in this method, which includes eight rollers 25 installed front and rear on one side of a box-shaped container 24, and a cutting device installed on the delivery side between the rollers. It consists of rotating blades 27 attached at regular intervals to a rotating shaft 26 parallel to the roller 25. Each sun roller 25 and rotating shaft 26 are driven to rotate by a motor with a speed reducer (not shown), and in the case of the roller 25, the rotating directions are opposite to each other so as to draw in the plant body. . Rotating axis 2
6 may be rotated in any direction. As shown in the figure, each Q-ra 25 is also formed with a circumferential groove 3L at a location corresponding to the rotary blade 27, into which the cutting edge of the G-rotary blade 27 is loosely fitted. As a result, the rotary blade 27 is l
” - It is possible to reliably cut the plant body without contacting the blade 25. 28 is a presser plate that can move forward and backward; 29 is a supply port; and 30 is a discharge port.
Immediately after delivery 17, it is cut by the same rotary blade and the wandering i
It is sequentially discharged from Jj[]30.

This cutting device rW'7I is designed to cut the plant sent out from between L1 and R with each rotary blade, but instead of the rotary blade, each cutting blade moves back and forth in the left and right direction in the figure, that is, it is a roller. Alternatively, a fixed blade whose cutting edge is fixed parallel to the roller with its cutting edge facing forward and backward, and a movable blade that moves back and forth in the forward and backward direction may be provided so that a fixed amount of the plant body is sent out from the roller. It is also possible to cut it every time. Figures 10 to 12 show other embodiments of the above-mentioned cutting device, and Figure 10 shows rotary blades 33 arranged between rollers 32 and arranged in a row, with the cutting edge of each rotary blade 33 connected to the rollers 32 on both sides. It is loosely fitted into the circumferential groove 34. Figure 1.1 shows the 10th
A hook 35 is provided protruding from the circumferential surface between the grooves of the roller 32 shown in the figure, so that a plant can be hooked and raked. In FIG. 12, circumferential grooves 37 are alternately formed on opposing rollers 36, and a rotary blade 38 is provided in each circumferential groove so as to protrude from the groove bottom. In another invention for preparing cultured sections, a cutting device is used in which cutting blades are combined so that the cutting edges are parallel, lattice-shaped, or concentric, and either one of the sterilized or sterilized plant is kept under sterile conditions. A method is provided for cutting a plant by pressing it under one another.

FIG. 13 shows an example of a cutting device used in this method, which consists of cutting blades 42 arranged in parallel on one side of a box-shaped container 4l with their cutting edges facing the other side.
On the other side, a presser foot vi43 is provided so as to be able to move forward and backward toward the cutting blade 42. 44 is a supply port, 45 is a discharge port, and after a suitable amount of the plant body supplied from the supply port 44 is stored in the container, the holding plate 43 pushes the plant body and presses it against the cutting edge, so that the plant body is pressed and cut. The cut plant body is discharged from the discharge port 45.

In this method, the cut plants tend to get stuck between the cutting blades, causing clogging. This problem can be solved by locating the cutting edge on a concave curved surface and gradually widening the interval between the cutting blades from the cutting edge toward the delivery side. FIG. 14 shows an example of this, in which the cutting edges of each cutting blade 49 provided in parallel on one side of a cylindrical container 48 are repositioned onto a spherical surface, and each blade is orthogonal to the spherical surface. 5
0 is a presser plate whose presser surface has a spherical shape that matches the spherical surface above. This allows the distance between the presser surface and the cutting edge to be kept constant during press-cutting, making it possible to cut without leaving any uncut edges in either the center or the periphery.
Each of the above-mentioned cutting devices is fixed and the plant is pushed in with a presser plate, but the cutting device may be made movable back and forth so as to cut the plant as it moves forward. In this case, the presser plate can be omitted.

Each of the above contents 111. ．． 19, 24, and 41, the plants may be supplied in a state suspended in water or a liquid medium, or only the plants may be supplied. In still another invention, a cutting device having a movable blade that rotates, oscillates or reciprocates is provided in a channel through which water or a liquid medium in which plants are suspended is provided, and the cutting device is provided with a movable blade that rotates, oscillates or reciprocates, and the plant body is flown while suspended in the water or liquid medium. The plants that are removed are cut by a movable blade and flowed downstream.

As the cutting device used in this method, for example, the above-mentioned first
, 6 to 13 and 16, especially the apparatus shown in FIGS.
The device shown in FIG. 12 is a preferred example.

FIG. 15 shows a typical example.

In this device, a presser plate may be used together, but it can also be omitted. According to the method shown in FIGS. 6 to 15 described above, cultured sections cut into thin pieces can be obtained, but these methods are not suitable for plants with unilateral buds, such as roses, miniature roses, carnesian suns, chrysanthemums, etc. Suitable for obtaining cultured sections from the body. There is no problem with such plants as long as the lateral buds are not damaged, but according to the method shown in Figs. 6 to 15, the lateral buds are rarely damaged and cultured slices with intact lateral buds can be easily mass-produced. The above-mentioned cutting devices are all horizontally oriented, but
It can be oriented vertically or tilted diagonally if necessary. Example 1 Cutting edge 5a of blade 5 and circle w. First, using the cutting device shown in Figures 1 and 2 with a gap of 2 am between the
Wrap it with an aluminum wheel while covering it, and then
It is placed in a crepe and sterilized with steam. Next, it is transported to a sterile room, and the alkali wheel is peeled off. After that, the shoot lily shoot apical culture obtained from the gun lily shoot 5-8 mm diameter is obtained.
From injection day 6, 60 small Illusfree balls were pushed in, and a weight was placed on top of it via a presser plate (not shown).
Culture slices were obtained by pressing and cutting on plate F rotating at a rotation speed of 35 rpm with a pressure of 00 g/ci.

This cultured section was added to the culture solution by i.p. ty:ya niger(
When culturing was performed in wet air (lOO%0,), 2360 roots with a diameter of 3 sl1 were obtained after 15 weeks of culture.

Example 2 Cylindrical lump of potato (diameter 28*II, height 65 mm)
was pressed with a presser foot pressure of 45 g/c, using various rotational speeds from jf & f ft, and cut with a cutting edge of 2fi. As shown in Figure 4, sections of uniform width were obtained at 100 to 300 rpm. ．． 1f Example 3 A cylindrical lump of the same potato as in Example 2 was cut as a test material with a blade rotation speed of 35 Orpm and a blade of $52 am using various charges. 200-...350g
Sections of uniform width were obtained at /ci.

Example 4 Using the cutting device shown in Figs. 16 to 18, a small L gun lily
Cut E roots and obtain cultured sections No. 1:. When this was subjected to camellia molding in the same manner as in Example 1, almost the same results as the Example beads were obtained.

Effects of the Invention According to the method of the present invention, cultured sections of plant tissues that can be cultured without causing necrosis even when subjected to tissue culture can be easily produced in large quantities.

In addition, clogging of the cutting device can be prevented by positioning the cutting edge of the cutting blade on a concave curved surface and by gradually widening the corner between the cutting blades from the cutting edge toward the delivery side. Then, cut the plant by floating it in water or a liquid medium and cutting it with a movable blade! Damage to the cultured section can be reduced compared to pressing it against a Ji device and cutting it.

[Brief explanation of drawings]

FIG. 1 is a longitudinal sectional view of the cutting device W used in the method of the present invention,
, FIG. 2 is a cross-sectional view of the same, FIG. 3 is an enlarged cross-sectional view taken along the line A-A in FIG. lVj of the rotating blade when
1) A graph examining the relationship between the number of rotations, section thickness, and 2) variation δ. Figure 5 is a graph examining the relationship with load. FIG. 7 is a schematic diagram of another example; FIG. 8 is a schematic diagram of another cutting device; FIG. 9 is a sectional view taken along line B-B of the same device; FIG. Schematic side view of another cutting device,
FIG. 11 is a schematic diagram of yet another example. FIG. 12 is a schematic diagram of another example. FIG. 13 is a schematic diagram of another cutting 11i device. FIG. 14 is a schematic diagram of another example. 16 is a front view of another embodiment of the device shown in FIG. 1, FIG. 17 is a plan view of cutting device 0 in the same device, and FIG.
The figure is a sectional view taken along the line B-B in FIG. 17.1. ．． 11.
19.24.41.51...Yong8 3.53 ・
・Discs 5, 55...Blade 6. 16.29.
58 ... Supply port 7 ... Plow board 14.2
7.33.38 ...Rotary blade l5, 22'. 28...
Holding plate 17.30...Discharge [''20...Fixed blade 21...Movable blade 25.32, 3G...Roller 34.37...Peripheral groove 35...Fufuku 56...Subesa ejector. Mitsui Petrochemical Industries Co., Ltd.

Claims (1)

[Claims] (1) A plate is mounted on a disc rotationally driven by a motor so that the cutting edge faces outward rather than radially inward, and there is an appropriate gap between the plate and the disc, and the plate is mounted under sterile conditions. A method for producing cultured sections of plant tissue, which comprises supplying a plant body that has been sterilized or sterilized under a rotary blade and cutting it into slices to obtain a large number of cultured sections for tissue culture. (2) The method for preparing a cultured section of plant tissue according to claim 1, wherein the cutting edge of the blade is arcuate and the cutting is performed by pulling. (3) The blade is removably attached to the disk with a set screw. (4) The method for producing a cultured section of a plant tissue according to claim 3, wherein the blade is attached to the disk via a spacer (5) The disk is placed in a container. (6) The method for preparing a culture section of a plant tissue according to claim 1, wherein the container and the container are made of metal and can be sterilized by heat. The plant according to claim 1, wherein the number is 100 to 400 rpm, and the pressure applied to the blade is adjusted within the range of 0 to 500 g/cm^2 depending on the plant body, plant organ, tissue piece, or part thereof that is the material for culture. (7) A method for preparing a cultured section of a plant tissue according to claim 1, wherein the plant is a bulb (8) Rotating between the plow plates arranged in parallel at appropriate intervals;
A cutting device equipped with a movable blade that swings or moves back and forth and a sterilized or sterilized plant are pressed against the other under aseptic conditions to plow the plant with a plow board, and the movable blade is used. A method for preparing cultured sections of plant tissues characterized by obtaining a large amount of cultured sections to be used for tissue culture by cutting with a machine. A cutting device consisting of a movable blade that moves or oscillates to cut the plant body in cooperation with a fixed blade, and a sterilized or sterilized plant body is pressed against the other under sterile conditions to cut the plant body. A method for producing cultured sections of plant tissues, characterized in that a large number of cultured sections are obtained for tissue culture by cutting (10) a plurality of rollers arranged in parallel at appropriate intervals;
A cutting device consisting of a movable blade that rotates around an axis parallel to the roller axis on the delivery side between the rollers, moves forward and backward toward the roller, or reciprocates in a plane parallel to the roller, and is sterilized or sterilized. A cultured section of a plant tissue, characterized in that by pressing one of the plant bodies against the other, the plant body is sent out between rollers, and cut with a movable blade to obtain a large amount of cultured sections for tissue culture. Preparation method (11) Preparation of a cultured section of plant tissue according to claim 10, wherein the movable blade is a rotary blade that rotates around an axis parallel to the axis of the roller, and the cutting edge of the movable blade fits loosely into a circumferential groove formed on the roller. Method (12) A cutting device consisting of a plurality of rollers arranged in parallel at appropriate intervals, a rotary blade arranged between the rollers in line with the rollers, and rotating around an axis parallel to the rollers, and a cutting device that is sterile or A method for preparing cultured sections of plant tissues, which comprises pressing one of the sterilized plants against the other under aseptic conditions to cut the plants to obtain a large number of cultured sections for tissue culture (13) 12. The method for preparing a cultured section of a plant tissue according to claim 12, wherein the rotary blade has a cutting edge loosely fitted into a circumferential groove formed in the roller. A cutting device consisting of rotary blades arranged to protrude in the radial direction in circumferential grooves alternately formed in the circumferential grooves and a sterilized or sterilized plant body are pressed against the other under sterile conditions to cut the plant body. A method for preparing a plant tissue culture section, characterized by obtaining a large number of culture sections for tissue culture by cutting (15) A hook protruding from the circumferential surface between the grooves of the roller for scraping the plant body. Item 11, 13 or 14
Method for preparing cultured sections of plant tissue (16) A cutting device in which cutting blades are combined so that the cutting edges are parallel, lattice-shaped, or concentric, and either one of the sterilized or sterilized plant body is placed under sterile conditions. A method for preparing a cultured section of a plant tissue, characterized in that the plant body is cut by pressing the plant toward the other side to obtain a large number of cultured sections for tissue culture (17) The cutting edge is positioned on a concave curved surface. Claim 12 wherein the interval between the cutting blades is gradually widened from the cutting edge toward the delivery side.
The method for preparing a cultured section of a plant tissue (18) as described in claim 17, wherein a presser plate is provided to press the plant body against the cutting edge, and the presser plate has a convex curved surface corresponding to the curved surface.
Method for preparing cultured sections of plant tissues described in (19) A cutting device having a movable blade that rotates, swings or reciprocates is provided in a channel through which water or a liquid medium in which a plant body is suspended is flown, A method for producing cultured sections of plant tissue, characterized by cutting a floating plant body with a movable blade to obtain a large number of cultured sections for tissue culture (20) The plant body has lateral buds. A method for preparing a cultured section of a plant tissue according to claims 8 to 19.