CN222348975U - Plant callus device is infected to agrobacterium - Google Patents

Abstract

The utility model belongs to the technical field of biology, and discloses an agrobacterium infection plant callus device which comprises an infection bottle body, wherein an infection bottle cover is connected with the top thread of the infection bottle body, a through hole is formed in the center of the top of the infection bottle cover, a breathable film is arranged at the through hole, a containing barrel is integrally formed in the center of the bottom of the infection bottle cover, the device simply and quickly realizes agrobacterium-mediated genetic transformation infection, solves the problem of damage caused by repeated movement of callus in the conventional agrobacterium-mediated genetic transformation infection process, also solves the problem of waste caused by easy pouring of callus after infection is completed, reduces the operation flow, ensures consistency of callus infection time, enables the agrobacterium-mediated genetic transformation to be simpler and more convenient, reduces movement of callus in the operation process, reduces the possibility of pollution, and is also suitable for large-scale callus infection and has wide application.

Description

Plant callus device is infected to agrobacterium

Technical Field

The utility model relates to the technical field of biology, in particular to an agrobacterium infection plant callus device.

Background

The agrobacterium-mediated genetic transformation technology is the most commonly used transgenic method at present, and has the advantages of simple and convenient operation, low cost, high transformation efficiency, genetic stability and the like. Infection is a critical step in genetic transformation, and in conventional infection, callus is placed in a triangular flask and then poured into suspension to infect for a period of time. After infection is completed, pouring out infection liquid, transferring the callus onto sterile filter paper by forceps, sucking the surface bacterial liquid of the callus, and simultaneously, blowing the surface bacterial liquid of the callus in an ultra-clean workbench for a short time, and inoculating the callus into a co-culture medium for a period of time. And (3) performing sterilization treatment after co-culture is finished, placing the callus in a triangular conical flask, cleaning the callus for multiple times by using sterile water added with antibiotics, or soaking the callus in a suspension culture medium added with antibiotics for a period of time to achieve a bacteriostatic effect, pouring out a cleaning solution, and transferring the callus onto sterile filter paper by using forceps to suck surface moisture. All operations were performed in an ultra clean bench.

The method has the main defects that 1, in the infection process and the co-culture sterilization process, forceps are used for moving the callus for many times, callus cells are easy to damage, 2, after the infection is finished and when waste liquid is poured in the co-culture sterilization cleaning process, the callus is easy to pour out of a vessel, so that waste of experimental materials is caused, 3, after the infection is finished and the co-culture sterilization is finished, the callus is required to be transferred from a triangular flask to sterile filter paper and dried on an ultra-clean workbench, the transfer operation process is complicated, and the possibility of increasing pollution is increased.

Disclosure of utility model

The utility model aims at the problems in the prior art, designs an agrobacterium infection plant callus device which can treat a large amount of callus at one time, avoids damage caused by moving the callus by using tweezers for multiple times in the operation process, and avoids waste of materials and increase of pollution probability.

The utility model is realized in the following way:

The utility model provides an agrobacterium infects plant callus device, includes infects the bottle, the top threaded connection who infects the bottle has the bottle lid that infects, the through-hole has been seted up at the top central authorities of infects the bottle lid, through-hole department is provided with the ventilated membrane, the bottom central authorities integrated into one piece who infects the bottle lid has a holding section of thick bamboo, the holding section of thick bamboo is in the upper portion in the bottle that infects, the bottom of holding section of thick bamboo with the opening threaded connection at the top of the section of thick bamboo that infects, the bottom of holding section of thick bamboo with the joint of the interior wall bottom of the bottle that infects, it is provided with the fungus liquid discharge port to infest bottle bottom one side, fungus liquid discharge port department is provided with manual valve.

In one embodiment of the utility model, a circle of annular protrusions are arranged at the top of the infection bottle cap and close to the periphery, and filter paper is clamped between the annular protrusions and the top surface of the infection bottle cap.

In one embodiment of the utility model, a handle is arranged on one side of the peripheral surface of the infection bottle body, and anti-skid patterns are formed on the peripheral surface of the infection bottle cap.

In one embodiment of the utility model, the upper part of the infection cylinder is surrounded by a stainless steel plate, the middle part is surrounded by a stainless steel filter screen, and the bottom is a circular stainless steel plate.

In one embodiment of the utility model, a circle of clamping grooves are formed in the center of the bottom of the inner wall of the infection bottle body, and the edges of the round stainless steel plates at the bottom of the infection cylinder are clamped with the clamping grooves.

Compared with the prior art, the utility model has the beneficial effects that:

The device simply and rapidly realizes the genetic transformation infection of agrobacterium mediation, solves the problem of damage caused by moving the callus for many times in the conventional agrobacterium mediation genetic transformation infection process, also solves the problem of waste caused by pouring the callus after the infection is finished, reduces the operation flow, ensures the consistency of the infection time of the callus, ensures that the agrobacterium mediation genetic transformation is simpler and more convenient, reduces the movement of the callus in the operation process, reduces the possibility of pollution, is also suitable for large-scale callus infection, and can be used for the infection of other explants of plants, thereby having wide application.

Drawings

In order to more clearly illustrate the technical solutions of the embodiments of the present utility model, the drawings that are needed in the embodiments will be briefly described below, it being understood that the following drawings only illustrate some examples of the present utility model and therefore should not be considered as limiting the scope, and other related drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.

FIG. 1 is a schematic view of the overall structure of the present utility model;

Fig. 2 is a schematic view of the structure of an infested bottle cap;

FIG. 3 is a schematic view of the structure of an infested bottle;

fig. 4 is a schematic structural view of an infestation cylinder.

The reference numerals indicate that 1, an infected bottle cap, 2, a through hole, 3, a ventilated membrane, 4, an annular bulge, 5, a containing cylinder, 6, an infected cylinder, 7, an infected bottle body, 8, a clamping groove, 9, a bacterial liquid outlet, 10 and a handle.

Detailed Description

For the purpose of making the objects, technical solutions and advantages of the embodiments of the present utility model more apparent, the technical solutions of the embodiments of the present utility model will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present utility model, and it is apparent that the described embodiments are some embodiments of the present utility model, but not all embodiments. All other embodiments, based on the embodiments of the utility model, which are apparent to those of ordinary skill in the art without inventive faculty, are intended to be within the scope of the utility model.

Thus, the following detailed description of the embodiments of the utility model, as presented in the figures, is not intended to limit the scope of the utility model, as claimed, but is merely representative of selected embodiments of the utility model. All other embodiments, based on the embodiments of the utility model, which are apparent to those of ordinary skill in the art without inventive faculty, are intended to be within the scope of the utility model.

It should be noted that like reference numerals and letters refer to like items in the following figures, and thus once an item is defined in one figure, no further definition or explanation thereof is necessary in the following figures.

In the description of the present utility model, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", etc. indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings are merely for convenience in describing the present utility model and simplifying the description, and do not indicate or imply that the apparatus or elements referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present utility model.

Furthermore, the terms "first," "second," and the like, are used for descriptive purposes only and are not to be construed as indicating or implying a relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defining "a first" or "a second" may explicitly or implicitly include one or more such feature. In the description of the present utility model, the meaning of "a plurality" is two or more, unless explicitly defined otherwise.

Examples

Referring to the drawings, 1-4 show, the utility model provides an agrobacterium infection plant callus device, which comprises an infection bottle body 7, wherein an infection bottle cap 1 is connected with the top thread of the infection bottle body 7, a through hole 2 is formed in the center of the top of the infection bottle cap 1, a breathable film 3 is arranged at the position of the through hole 2, a containing barrel 5 is integrally formed in the center of the bottom of the infection bottle cap 1, the containing barrel 5 is positioned at the upper part in the infection bottle body 7, the bottom of the containing barrel 5 is connected with the opening thread at the top of the infection barrel 6, the bottom of the containing barrel 5 is clamped with the bottom of the inner wall of the infection bottle body 7, a bacterial liquid outlet 9 is arranged at one side of the bottom of the infection bottle body 7, and a manual valve is arranged at the bacterial liquid outlet 9.

In one embodiment of the utility model, further, a circle of annular protrusions 4 is arranged at the top of the infection bottle cap 1 and near the periphery, and filter paper is clamped between the annular protrusions 4 and the top surface of the infection bottle cap 1.

In one embodiment of the utility model, further, a handle 10 is arranged on one side of the peripheral surface of the infection bottle body 7, and anti-skid patterns are formed on the peripheral surface of the infection bottle cap 1.

In one embodiment of the utility model, further, the upper part of the infection cylinder 6 is surrounded by a stainless steel plate, the middle part is surrounded by a stainless steel filter screen, and the bottom part is a circular stainless steel plate.

In one embodiment of the utility model, a circle of clamping grooves 8 are arranged at the center of the bottom of the inner wall of the infection bottle body 7, and the edges of the round stainless steel plates at the bottom of the infection cylinder 6 are clamped with the clamping grooves 8.

Specifically, the invasion bottle cap 1 and the containing cylinder 5 integrally formed at the bottom of the invasion bottle cap are supported by a PP material, the invasion bottle body 7 is made of high-temperature transparent glass material (high-whiteness glass), the invasion cylinder 6 is made of a stainless steel filter screen and a stainless steel plate by welding, and all parts can be sterilized by adopting a high-temperature high-pressure mode;

In the use process, after the callus is placed in the infection cylinder 6 and the opening part of the callus is in threaded connection with the bottom of the accommodating cylinder 5 at the bottom of the infection bottle cap 1, the whole is placed in the infection bottle 7, the infection bottle 7 is poured with agrobacterium invasion solution along the edge in advance, the agrobacterium invasion solution enters into the soaked callus through a stainless steel mesh hole of the infection cylinder 6, the infection bottle cap 1 is tightly covered with the top threaded connection of the infection bottle 7, the bottom of the infection cylinder 6 is clamped with a clamping groove 8 arranged at the bottom of the inner wall of the infection bottle 7, the whole stability of the device is improved, the ventilation membrane 3 arranged at the through hole 2 at the top of the infection bottle cap 1 is used for increasing the ventilation property in the device, bacteria and dust can be isolated, the agrobacterium invasion solution can be discharged through a bacterial solution discharge port 9 arranged at the bottom of one side of the infection bottle 7 after the soaking is completed, the callus inside the device is effectively prevented from being led out to cause operation accidents, and the waste of materials is caused;

In addition, through set up round annular protruding 4 around the top of invasion bottle lid 1, through the aseptic filter paper of card in annular protruding 4, after the callus soaked, through the device inversion, the callus can get into smoothly in holding section of thick bamboo 5 and do not keep in to make the residual agrobacterium of callus surface invade the dye liquor accessible ventilated membrane 3 by aseptic filter paper absorption, need not to press from both sides many times through tweezers and get the condition that the callus leads to tissue damage and infection's risk to increase takes place.

In conclusion, the device simply and rapidly realizes agrobacterium-mediated genetic transformation infection, solves the problem of damage caused by repeated movement of callus in the conventional agrobacterium-mediated genetic transformation infection process, also solves the problem of waste caused by easy dumping of the callus after infection is completed, reduces the operation flow, ensures the consistency of the infection time of the callus, ensures that the agrobacterium-mediated genetic transformation is simpler and more convenient, reduces the movement of the callus in the operation process, reduces the possibility of pollution, is also suitable for large-scale callus infection, and can be used for infection of other explants of plants, thereby having wide application.

In the embodiment of the utility model, the height of the infection bottle body 7 is 120+/-0.1 mm, the diameter of the bottom of the infection bottle body 7 is 140+/-0.1 mm, the diameter of the infection bottle cap 1 is 132+/-0.1 mm, the outer diameter of the infection bottle cap 1 is 134+/-0.1 mm, the inner diameter is 130+/-0.1 mm, the height is 10+/-0.1 mm, the diameter of the through hole 2 is 50+/-0.1 mm, the diameter of the ventilation film 3 is 49+/-0.1 mm, the aperture is 0.22 mu m, the width of the annular bulge 4 is 3+/-0.1 mm, the distance between the annular bulge 4 and the infection bottle cap 1 is 6+/-0.1 mm, the diameter of the accommodating cylinder 5 is 102+/-0.1 mm, the height is 30+/-0.1 mm, the diameter of the infection cylinder 6 is 100+/-0.1 mm, and the height is 84+/-0.1 mm, the stainless steel filter screen comprises an upper stainless steel plate 10+/-0.1 mm high, a middle stainless steel filter screen 74+/-0.1 mm high, a stainless steel filter screen aperture 3+/-0.1 mm, a clamping groove 8 at the bottom with a height of 2+0.1mm, a distance from an infected bottle body 7 to 19+0.1mm, a width of 1+0.1mm, a fungus liquid discharge port 9 with a diameter of 8+0.1mm, a distance from the bottom of 2+/-0.1 mm, a handle 10 at the outer side of the infected bottle body 7 with a thickness of 8+/-0.1 mm, a height of 70+/-0.1 mm, an inner diameter distance from the infected bottle body 7 to 50+/-0.1 mm, and screw thread heights in the embodiment of 9+0.1mm, a bottle cap with a thickness of 1+0.1mm and a bottle bottom with a thickness of 5+0.1mm.

The working principle of the plant callus device infected by agrobacterium is as follows:

1. Sterilizing the whole set of device in a high-pressure high-temperature sterilizing pot, taking out, and opening in an ultra-clean workbench;

2. Placing a plurality of sterile filter papers on the inner side of the annular bulge 4, placing the callus in an infection cylinder 6, and pouring agrobacterium infection liquid into the edge of an infection bottle body 7;

3. The method comprises the steps of (1) screwing an infection bottle cap 1 with an infection bottle body 7 through threads, screwing a containing cylinder 5 with an infection cylinder 6 through threads, and starting infection;

4. After infection is finished, the bacterial liquid outlet 9 is opened to discharge bacterial liquid, if residual bacterial liquid exists, the whole device can be slightly inclined, so that the residual bacterial liquid is conveniently discharged;

5. Inverting the whole set of device to transfer the callus into the accommodating cylinder 5, sucking the bacterial liquid on the surface of the callus by using sterile filter paper placed between the annular bulge 4 and the infection bottle cap 11, unscrewing the infection bottle cap 1, taking out the whole set of device, the accommodating cylinder 5 and the infection cylinder 6, and blowing for 3-8 minutes by using sterile wind to fully remove the agrobacterium infection liquid;

6. Transferring the callus to a co-culture medium for 2-3d dark culture;

7. after co-cultivation is finished, the set of devices are used for washing or soaking;

8. After the water washing or soaking is finished, the operation of the step 5 is performed to suck the water and blow;

9. The callus is placed in a screening medium for screening.

The above is only a preferred embodiment of the present utility model, and is not intended to limit the present utility model, but various modifications and variations can be made to the present utility model by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present utility model should be included in the protection scope of the present utility model.

Claims (5)

1. The utility model provides an agrobacterium infects plant callus device, its characterized in that, including infesting bottle (7), the top threaded connection who infects bottle (7) has infects bottle lid (1), the through-hole (2) have been seted up at the top central authorities that infects bottle lid (1), through-hole (2) department is provided with ventilated membrane (3), the bottom central authorities that infects bottle lid (1) integrated into one piece has holding section of thick bamboo (5), holding section of thick bamboo (5) are in the upper portion in infects bottle (7), the bottom of holding section of thick bamboo (5) with the opening threaded connection at the top of invading section of thick bamboo (6), the bottom of holding section of thick bamboo (5) with the joint of the inner wall bottom of invading bottle (7), one side of the bottom of invading bottle (7) is provided with fungus liquid discharge port (9), fungus liquid discharge port (9) department is provided with manual valve.

2. The agrobacterium infection plant callus device according to claim 1, wherein a circle of annular protrusions (4) is arranged at the top of the infection bottle cap (1) and close to the periphery, and filter paper is clamped between the annular protrusions (4) and the top surface of the infection bottle cap (1).

3. The agrobacterium infection plant callus device according to claim 2, wherein a handle (10) is arranged on one side of the peripheral surface of the infection bottle body (7), and anti-skidding patterns are formed on the peripheral surface of the infection bottle cap (1).

4. An agrobacterium infection plant callus apparatus according to claim 3, wherein the upper part of the infection cylinder (6) is provided with stainless steel plates around, the middle part is provided with a stainless steel filter screen around, and the bottom is provided with a circular stainless steel plate.

5. The agrobacterium infection plant callus device according to claim 4, wherein a circle of clamping groove (8) is arranged at the center of the bottom of the inner wall of the infection bottle body (7), and the edge of the circular stainless steel plate at the bottom of the infection cylinder (6) is clamped with the clamping groove (8).