Portable pressure osmometer and pressure osmosis method

Abstract

The present invention discloses a portable pressure osmometer and a pressure osmosis method, which are used for pressure osmosis of living plants, and include: a sample processing chamber, an osmotic pressure regulating device and a gas pipeline; the sample processing chamber is provided with a sample inlet and a liquid adding port for adding osmotic liquid; the sample processing chamber is also provided with a first sealing device for sealing the sample inlet and a second sealing device for sealing the liquid adding port, wherein a channel for the stem and / or leaf of the living plant to pass through is provided in the first sealing device; one end of the gas pipeline is connected to the osmotic pressure regulating device, and the other end is connected to the sample processing chamber; the osmotic pressure regulating device can output a gas with a constant pressure, and the gas is used to regulate the osmotic pressure of the osmotic liquid in the sample processing chamber. The portable pressure osmometer and a pressure osmotic method are used for pressure osmosis of living plants outdoors at a certain constant pressure, thereby accelerating the osmotic efficiency and shortening the osmotic cycle.

Description

Technical Field

[0001] The invention belongs to the technical field of plant genetic engineering equipment, and in particular relates to a portable pressure osmosis instrument and a pressure osmosis method. Background Art

[0002] Plant transient transformation refers to the process of introducing exogenous genes into the genome of recipient plant cells through a certain method so that they are expressed in the cells, thereby enabling rapid study of gene function.

[0003] For outdoor living plants, currently, the infection liquid is mainly applied evenly on the surface of the outdoor living plants to penetrate into the interior of the living plants, which takes a long time to penetrate and has low efficiency; or the infection liquid is penetrated into a local area of ​​the leaves using a syringe, which has a narrow penetration area and is easy to damage the leaves. It is impossible to perform pressure penetration on outdoor living plants. Summary of the invention

[0004] The purpose of the present invention is to provide a portable pressure penetration instrument and a pressure penetration method to solve the problem that it is impossible to perform pressure penetration on outdoor living plants.

[0005] The technical solution is as follows:

[0006] The portable pressure osmometer of the present invention is used for pressure osmosis of living plants, comprising:

[0007] Sample processing chamber, osmotic pressure adjustment device and gas pipeline;

[0008] The sample processing chamber is a hollow cavity, provided with a sample inlet and a liquid adding port for adding permeate; the sample processing chamber is also provided with a first sealing device for sealing the sample inlet and a second sealing device for sealing the liquid adding port, and a channel for the stem and / or leaf of the living plant to pass through is provided in the first sealing device;

[0009] One end of the gas pipeline is connected to the osmotic pressure regulating device, and the other end is connected to the sample processing chamber;

[0010] The osmotic pressure regulating device can output a gas with a constant pressure, and the gas is used to regulate the osmotic pressure of the permeate in the sample processing chamber.

[0011] In one embodiment, the first closing device includes an injection port plug and a plug cover, the injection port plug is provided with the channel, the injection port plug includes an inner plug installed in the injection port in an interference fit manner and an outer plug located outside the injection port, the plug cover is a hollow cylinder with openings at both ends, the inner wall of the plug cover is tightly matched with the outer wall of the outer plug, and under the extrusion of the injection port and the plug cover, the injection port plug can shrink in the radial direction to close the channel.

[0012] In one embodiment, the injection port plug is provided with a first groove extending through the injection port plug in the axial direction, and the first groove forms the channel.

[0013] In one embodiment, the plug cover is provided with a second groove extending axially therethrough, and the second groove is configured to allow the stem and / or leaves of the living plant to pass through the plug cover.

[0014] In one embodiment, the inner wall of the plug cover and the outer wall of the outer plug are both provided with mutually coupled threads.

[0015] In one embodiment, the radial dimension of the inner plug gradually increases from an end away from the center of the injection port plug to an end adjacent to the center of the injection port plug.

[0016] In one embodiment, an injection port cover is further provided. The injection port cover is cylindrical and one end is closed. The injection port cover is screwed to the injection port thread.

[0017] In one of the embodiments, a valve is provided on the gas pipeline.

[0018] The pressure penetration method of the present invention is used to perform pressure penetration on living plants, comprising:

[0019] Passing a living material through a sealing device, placing it into a sample processing chamber from a sample inlet, and sealing the sample inlet, wherein the sealing device is used to seal the sample inlet of the sample processing chamber, and the living material is a stem and / or leaf of the living plant;

[0020] injecting permeate into the sample processing chamber from a liquid filling port of the sample processing chamber and sealing the liquid filling port;

[0021] Setting the pressure of the gas output by the osmotic pressure regulating device;

[0022] The gas outputted from the osmotic pressure regulating device is inputted into the sample processing chamber to perform pressure osmosis on the living material.

[0023] In one embodiment, the sealing device comprises an injection port plug and a plug cover, and the step of passing the living material through the sealing device, placing the living material into the sample processing chamber from the injection port, and sealing the injection port comprises:

[0024] Passing the living material through the injection port plug and then placing it into the sample processing chamber;

[0025] squeezing the injection port plug into the injection port to close the injection port;

[0026] The plug cover is used to compress the portion of the injection port plug exposed at the injection port to improve the sealing effect.

[0027] The technical solution provided by the present invention has the following advantages and effects:

[0028] A channel for the stem and / or leaf of a living plant (referred to as living material) to pass through is provided on the first sealing device that seals the sample inlet. The first sealing device can seal the sample inlet when the living material passes through. The osmotic pressure regulating device then outputs a gas at a constant pressure to regulate the osmotic pressure of the permeate in the sample processing chamber. The pressure permeation can be performed on the living plants outdoors at a certain constant pressure, thereby accelerating the permeation efficiency and shortening the permeation cycle. BRIEF DESCRIPTION OF THE DRAWINGS

[0029] Figure 1 is a schematic diagram of a portable pressure permeameter provided by an embodiment of the present invention;

[0030] Figure 2 is a schematic diagram of another portable pressure osmometer provided by an embodiment of the present invention;

[0031] Figure 3 It is a schematic diagram of the portable pressure permeameter used in the scenario of pressure permeation of in vitro materials;

[0032] Figure 4 is a schematic flow chart of a pressure penetration method provided by an embodiment of the present invention;

[0033] Figure 5 yes Figure 4 A schematic flow chart of step S100 in the embodiment.

[0034] Description of reference numerals:

[0035] 10. Sample processing chamber, 11. Liquid adding port, 12. Sample injection port, 13. First closing device, 14. Sample injection port plug, 14a. First groove, 15. Plug cover, 15a. Second groove, 16. Sample injection port cover, 20. Osmotic pressure regulating device, 22. Piston, 30. Gas pipeline. DETAILED DESCRIPTION

[0036] In order to facilitate the understanding of the present invention, specific embodiments of the present invention will be described in more detail below with reference to the accompanying drawings.

[0037] Unless otherwise specified or defined, the "first, second..." used in this article is merely used to distinguish names and does not represent a specific quantity or order.

[0038] Unless specifically stated or defined otherwise, the term "and / or" as used herein includes any and all combinations of one or more of the associated listed items.

[0039] It should be noted that, in this document, “fixed to” or “connected to” may mean directly fixing or connecting to an element, or indirectly fixing or connecting to an element.

[0040] In view of the problem that it is currently impossible to perform pressure penetration on outdoor living plants to improve penetration efficiency and shorten penetration time, the present invention provides a portable pressure penetration instrument, which provides a channel for the stems and / or leaves of living plants (referred to as living materials) to pass through on the sealing device of the sealed sample inlet. During pressure penetration, the sample inlet is sealed when the living material passes through the sealing device and is placed in the sample processing chamber, and the osmotic pressure of the permeate in the sample processing chamber is adjusted by outputting gas at a constant pressure through the osmotic pressure regulating device. The pressure penetration of living plants is achieved under a certain constant pressure, the penetration efficiency is accelerated, and the penetration cycle is shortened.

[0041] Figure 1 The portable pressure osmometer of this embodiment is shown, which mainly includes a sample processing chamber 10, an osmotic pressure regulating device 20 and a gas pipeline 30, one end of the gas pipeline 30 is connected to the osmotic pressure regulating device 20, and the other end is connected to the sample processing chamber 10, for transmitting the constant pressure gas output by the osmotic pressure regulating device 20 to the sample processing chamber 10.

[0042] Wherein, the sample processing chamber 10 is a hollow cavity, and the shape is a sphere, but it is not limited to a sphere, and can also be various shapes such as a cylinder, a cube, a cone, a flat body, etc. According to the use environment requirements, the sample processing chamber 10 can be made of glass, plastic, metal, and composite materials. The osmotic pressure regulating device 20 is a small pneumatic tool (such as a nail gun, a spray gun, etc.) that can output a constant air pressure. These small pneumatic tools are equipped with a control panel for adjusting the pressure of the output gas, and a piston 22 is installed inside. When the piston 22 is pushed, the gas at a constant pressure is pushed into the gas pipeline 30. In order to facilitate the pressure penetration operation of living plants outdoors, a battery-driven gas tool or a manual pressure regulating tool can be purchased, which is particularly suitable for use in an environment without a convenient power supply or air compressor. Among them, the battery-driven gas tool uses a built-in battery to drive a small electric compressor, which can compress air or other gases and output them at different pressures; the manual pressure regulating tool can compress air through a manual pump or other mechanical device, store it in a built-in small pressure container, and output it at different pressures.

[0043] The sample processing chamber 10 is provided with a liquid adding port 11 for adding permeate, and for placing the stems and / or leaves of a living plant into the sample processing chamber 10 through the injection port 12. The injection port 12 is sealed with a first sealing device 13, and the liquid adding port 11 is sealed with a second sealing device (not shown in the figure). In this embodiment, for ease of operation, the injection port 12 and the liquid adding port 11 are different entrances. In one example, the injection port 12 is also used as the liquid adding port 11, and the permeate is first injected into the sample processing chamber 10 from the injection port 12, and then the stems and / or leaves of the living plant are placed into the sample processing chamber 10 through the injection port 12. In this example, the first sealing device 13 and the second sealing device are the same device.

[0044] In order to achieve pressure penetration of living plants, the first sealing device 13 is different from the commonly used wooden stoppers and glass stoppers for sealing bottle mouths. It is necessary to open a channel in the first sealing device 13 for the stems and / or leaves of the living plants to pass through. With the help of this channel, living materials such as stems and / or leaves do not need to be separated from the living plants, and the living plants can be directly pressure penetrated in a sealed state.

[0045] In the present embodiment, the first closing device 13 includes an injection port plug 14 and a plug cover 15. The injection port plug 14 is made of a soft material, such as rubber, silicone, cork, etc. Under radial extrusion, the injection port plug 14 can shrink in the radial direction. The injection port plug 14 is provided with a first groove 14a that runs through the front and back along its axial direction, and the first groove 14a is the above-mentioned channel for the stem and / or leaves of the living plant to pass through. The stem of the living plant can be easily inserted into the first groove 14a, and can also be applied to the stems of living plants of different diameters. The stems and leaves in front of the injection port plug 14 (towards the injection port) can extend into the sample processing chamber 10. It is also achieved that the stems and / or leaves of the living plant pass through the injection port plug 14 and are placed in the sample processing chamber 10. The injection port plug 14 is installed in the injection port 12 by means of interference fit. Under the pressure of the inner wall of the injection port 12, the injection port plug 14 will shrink in the radial direction to close the first groove 14a.

[0046] In one example, the injection port plug has a front-to-back through-channel dug in the center along the axial direction, and the stem and / or leaves of the living plant are gathered and then passed through the channel and then extended into the sample processing chamber. Similarly, when installed in the injection port, the injection port plug will shrink in the radial direction under the pressure of the inner wall of the injection port to close the channel.

[0047] In this embodiment, only part of the injection port plug 14 is squeezed and installed into the injection port 12, that is, the injection port plug 14 includes two parts: one part is an inner plug installed in the injection port 12, and the other part is an outer plug located outside the injection port 12. In this way, after the pressure penetration is completed, it is not only convenient to pull the injection port plug 14 out of the injection port, but also not easy to damage the stems and / or leaves of the living plant. However, in this case, when the injection port is closed, the inner plug shrinks and the outer plug expands, and the outer plug may cause pulling deformation to the inner plug, affecting the sealing effect. In order to improve the sealing effect, a plug cover 15 is used to compress the outer plug. Specifically, compared with the injection port plug 14 made of soft material, the plug cover 15 is made of metal or rigid material, with openings at both ends, and is a hollow cylinder. The stems and / or leaves of the living plant can directly pass through the cavity in the plug cover 15. When the plug cover 15 is installed, the inner wall of the plug cover 15 forms a tight fit with the outer wall of the outer plug. The above-mentioned channel also exists in the outer plug. Under the pressure of the plug cover 15, the outer plug will shrink in the radial direction to close the channel in the outer plug. In this embodiment, the inner surface of the plug cover 15 is provided with a thread, and the outer surface of the outer plug is also provided with a matching thread. The plug cover 15 is installed on the outer plug by screwing. However, it is not limited to this assembly method. In one example, the plug cover 15 is made into an annular clamp, and the annular clamp is sleeved on the outer plug. The size of the through hole of the annular clamp is adjusted by the adjustment device (such as a screw mechanism) on the annular clamp, thereby compressing the radial size of the outer plug. The above-mentioned annular clamp is often used to surround and tighten tubular objects (such as hoses, pipes, etc.), which will not be repeated here.

[0048] In order to facilitate the passage of the living material through the plug cover 15, a second groove 15a is axially dug through the plug cover 15. By designing the first groove 14a and the second groove 15a, the sealing between the living material passing through the injection port plug 14 and the plug cover 15 is achieved, and the living material can be pressure-permeated.

[0049] Compared with setting a through hole in the center of the injection port plug 14 for the living material to pass through, the first groove 14a is adopted, so that the upper and lower ends of the first groove 14a formed on the injection port plug 14 can be more easily squeezed and fitted when squeezed by the injection port; and considering the sealing performance, the size of the through hole set in the center is limited, and it is difficult to adapt to the stems of living plants of different sizes, and the squeezing force is dispersed on the inner wall of the injection port plug 14. Even if there is a stem of a living plant in the through hole in the center, it may be difficult to make the through hole in the center fit with the stem of the living plant.

[0050] In one example, if Figure 2 As shown, the injection port plug 14 is in the shape of a cone, and no thread is arranged on the injection port plug 14. The injection port plug 14 is squeezed into the injection port by screwing the plug cover 15 with the thread of the injection port 12.

[0051] In one example, the length of the plug cover 15 is greater than that of the outer plug and is in a step shape. The front end is screwed to the injection port 12, and the rear end is matched with the outer plug. The radial dimension of the front end is greater than the radial dimension of the rear end. Through the screw connection between the plug cover 15 and the injection port 12, when the injection port plug 14 is pushed forward (towards the injection port 12) and pressed into the injection port 12, the extrusion force is increased, which is convenient for operation.

[0052] On the basis of providing the plug cover 15 to enhance the sealing effect, the inner plug in this embodiment is designed to be a truncated cone, that is, the radial dimension of the inner plug gradually increases from the end away from the center of the injection port plug 14 to the end adjacent to the center of the injection port plug 14, thereby reducing the difficulty of installing the injection port plug to the injection port.

[0053] For indoor in vitro materials, negative pressure methods are currently mostly used to penetrate the infection liquid into the material. However, this method generally requires pre-treatment, such as ultrasonic treatment, to form tiny holes in the leaves, so as to improve efficiency. The single negative pressure method is not efficient and is mainly used for pressure penetration of in vitro materials in the laboratory.

[0054] In this embodiment, in order to improve the application scenarios of the portable pressure osmometer, a function of performing pressure osmotic experiments on in vitro materials is added, such as Figure 3 As shown, threads are provided on the outer surface of the injection port 12 , and an injection port cover 16 is also designed. The injection port cover 16 is cylindrical and one end is closed. The injection port cover 16 is screwed to the injection port 12 to close the injection port 12 .

[0055] That is, when pressure infiltration is performed on a living material, the injection port plug 14 and the plug cover 15 are used to seal the injection port; when pressure infiltration is performed on an in vitro material, the injection port cover 16 is used to seal the injection port.

[0056] In order to prevent the permeate from flowing back to the osmotic pressure regulating device, in one example, a valve is further provided on the gas pipeline.

[0057] When using the portable pressure osmometer of the present embodiment, the injection port plug and the plug cover are installed on the living material (generally on the stem of a woody plant), and the living material passes through the injection port 12 and is placed in the sample processing chamber 10. The injection port 12 is plugged with the injection port plug 14 and the plug cover 15 is installed on the injection port plug 14. Press zero on the control panel of the osmotic pressure regulating device, and the piston returns to the leftmost end. The sample processing chamber 10 is placed vertically, and the permeate is injected into the sample processing chamber from the liquid filling port 11, and then the liquid filling port 11 is closed. Through the control panel, the electric power is regulated to achieve the effect of fixed pressure. Pull the trigger, and the pressure is transmitted to the sample processing chamber 10 to adjust the osmotic pressure of the permeate. By changing the osmotic pressure of the permeate, the infection liquid can be directly infiltrated into the plant cell body, realizing the instantaneous transformation process on the living plant. This method can increase the permeability of the cell membrane, making it easier for exogenous DNA to enter the plant cell, thereby realizing the instantaneous transformation of exogenous genes in living cells and improving the efficiency of instantaneous transformation.

[0058] After the pressure osmosis process is completed, the second sealing device can be removed and the liquid filling port can be used as an exhaust port; the gas pipeline can also be disconnected from the sample processing chamber or the gas pipeline can be disconnected from the osmotic pressure regulating device to reduce the pressure in the sample processing chamber. Then, the first sealing device is removed from the sample processing chamber, the living plant is pulled out of the sample processing chamber, and the first sealing device is removed from the living plant.

[0059] In summary, this embodiment designs a channel in the injection port plug so that the living material can pass through the injection port plug, and assembles a plug cover on the injection port plug. Under the extrusion of the injection port and the plug cover, the injection port plug can be contracted in the radial direction to close the channel, so that the pressure penetration of the living plant can be performed in the sample processing chamber. By applying pressure, it can penetrate into the plant tissue more quickly, improving the efficiency of plant infection. In addition, an osmotic pressure regulating device that can output a constant gas pressure is adopted, so that the penetration of the living material under constant pressure can be achieved.

[0060] Under the concept of performing pressure penetration of living materials under constant pressure to improve penetration efficiency, the present invention also provides a pressure penetration method, such as Figure 4 As shown, the specific steps include:

[0061] Step S100: placing the living material through the sealing device, into the sample processing chamber from the injection port, and sealing the injection port;

[0062] The sealing device is used to seal the sample inlet of the sample processing chamber, and the living material is the stem and / or leaf of a living plant, and in this embodiment, it is the stem tip and / or leaf with petiole of the living plant;

[0063] Step S200: injecting permeate into the sample processing chamber from the liquid filling port of the sample processing chamber and sealing the liquid filling port;

[0064] Step S300: setting the gas pressure output by the osmotic pressure regulating device;

[0065] Step S400: inputting the gas outputted by the osmotic pressure regulating device into the sample processing chamber to perform pressure osmosis on the living material.

[0066] Optionally, the sealing device includes an injection port plug and a plug cover, such as Figure 5 As shown, step S100 specifically includes:

[0067] Step S110: placing the living material through the injection port plug into the sample processing chamber;

[0068] Step S120: Squeeze the injection port plug into the injection port to close the injection port;

[0069] Step S130: Use the plug cover to press the portion of the injection port plug exposed at the injection port to improve the sealing effect.

[0070] Specifically, in this embodiment, the specific functions of the above-mentioned pressure penetration method can refer to the corresponding description in the above-mentioned portable pressure penetration instrument, and will not be repeated here.

[0071] The above embodiments are not exhaustive enumerations of the present invention, and there may be multiple other implementations not listed. Any replacement and improvement made without violating the concept of the present invention shall fall within the protection scope of the present invention.

Claims

1. A portable pressure osmometer for pressure osmosis of living plants, characterized in that: include: Sample processing chamber, osmotic pressure adjustment device and gas pipeline; The sample processing chamber is a hollow cavity, provided with a sample inlet and a liquid adding port for adding permeate; the sample processing chamber is also provided with a first sealing device for sealing the sample inlet and a second sealing device for sealing the liquid adding port, and a channel for the stem and / or leaf of the living plant to pass through is provided in the first sealing device; One end of the gas pipeline is connected to the osmotic pressure regulating device, and the other end is connected to the sample processing chamber; The osmotic pressure regulating device can output a gas with a constant pressure, and the gas is used to regulate the osmotic pressure of the permeate in the sample processing chamber; The first sealing device comprises an injection port plug and a plug cover, wherein the injection port plug is provided with the passage, wherein the injection port plug comprises an inner plug installed in the injection port in an interference fit manner and an outer plug located outside the injection port, wherein the plug cover is a hollow cylinder with openings at both ends, wherein the inner wall of the plug cover is tightly matched with the outer wall of the outer plug, and under the extrusion of the injection port and the plug cover, the injection port plug can be contracted in a radial direction to close the passage; The injection port plug is provided with a first groove penetrating along the axial direction, and the first groove forms the channel; The plug cover is provided with a second groove which penetrates axially, and the second groove is configured to allow the stem and / or leaves of the living plant to pass through the plug cover.

2. The portable pressure osmometer according to claim 1, characterized in that: The inner wall of the plug cover and the outer wall of the outer plug are both provided with mutually coupled threads.

3. The portable pressure osmometer according to claim 1, characterized in that: The radial dimension of the inner plug gradually increases from an end away from the center of the injection port plug to an end adjacent to the center of the injection port plug.

4. The portable pressure osmometer according to claim 1, characterized in that: A sample injection port cover is also provided, which is cylindrical and has one end closed, and is screwed to the sample injection port thread.

5. The portable pressure osmometer according to claim 1, characterized in that: The gas pipeline is provided with a valve.

6. A pressure penetration method based on the portable pressure penetration instrument as claimed in any one of claims 1 to 5, used for pressure penetration of living plants, characterized in that: include: Passing a living material through a sealing device, placing it into a sample processing chamber from a sample inlet, and sealing the sample inlet, wherein the sealing device is used to seal the sample inlet of the sample processing chamber, and the living material is a stem and / or leaf of the living plant; injecting permeate into the sample processing chamber from a liquid filling port of the sample processing chamber and sealing the liquid filling port; Setting the pressure of the gas output by the osmotic pressure regulating device; The gas outputted from the osmotic pressure regulating device is inputted into the sample processing chamber to perform pressure osmosis on the living material.

7. The pressure infiltration method according to claim 6, characterized in that: The sealing device comprises an injection port plug and a plug cover, and the living material is passed through the sealing device, placed into the sample processing chamber from the injection port, and the injection port is sealed, comprising: Passing the living material through the injection port plug and then placing it into the sample processing chamber; squeezing the injection port plug into the injection port to close the injection port; The plug cover is used to compress the portion of the injection port plug exposed at the injection port to improve the sealing effect.

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