Device for instantly infecting radix tetrastigme with agrobacterium tumefaciens

Abstract

The utility model discloses an instantaneous agrobacterium infection device for radix tetrastigme, belongs to the technical field of plant infection, and aims to solve the problem that an existing infection device can only treat a single sample. Comprising an infection assembly and a net bag, the infection assembly comprises a bracket, a tray, containers and upper covers, the multiple containers are placed on the tray, the multiple upper covers cover the multiple containers in a one-to-one correspondence mode, holes are formed in the upper covers, the bracket is placed on the multiple upper covers, the bracket is provided with an inner circular ring, bag openings are formed in the upper end and the lower end of the net bag, and tightening pull ropes are arranged at the bag openings. Tetrastigma hemsleyanum is planted in the flowerpot, the flowerpot is arranged in an inverted mode, the opening end of the flowerpot extends into the net bag, the two tightening pull ropes are tightened, the flowerpot is placed on the bracket, and a plurality of branches of the Tetrastigma hemsleyanum penetrate out of a bag opening in the lower end of the net bag and correspondingly extend into the containers through the holes. Different bacterial strain infection solutions are arranged in the plurality of containers, so that multiple treatments can be simultaneously infected, the infection process is shortened, and the time is saved.

Description

TECHNICAL FIELD

[0001] The utility model belongs to the technical field of plant infection, especially relates to a radix tetrastigme agrobacterium transient infection device. BACKGROUND

[0002] Radix tetrastigme is a rare medicinal plant in China. In recent years, pharmacological studies have shown that radix tetrastigme has anti-inflammatory and antipyretic, antibacterial and antiviral, immune homeostasis regulation and anti-tumor functions. At the same time, the tubers of radix tetrastigme are rich in terpenes, flavonoids, phenolic acids and polysaccharides and other secondary metabolites, making it a good model plant for studying plant secondary metabolites. Due to the problem of regenerating seedlings in the tissue culture of radix tetrastigme, it is an ideal transformation method to directly infect the leaves or tubers of radix tetrastigme with agrobacterium by transient transformation method. In this method, agrobacterium carries certain enzyme genes or other functional genes, which can directly verify the function of the gene according to the phenotype of the organ or the enrichment amount of related secondary metabolites.

[0003] The leaves of radix tetrastigme are the main organs for genetic transformation. Due to long-term vegetative propagation, the genotype of radix tetrastigme is relatively hybrid, resulting in large genetic differences between plant individuals. Therefore, comparison of different treatments on the same plant is an ideal experimental scheme, which can effectively reduce the error caused by individual differences between different plants. Radix tetrastigme has many branches, long branches and infinite growth characteristics, so when carrying out large-scale genetic transformation, not only the multi-branch characteristics need to be used to meet the simultaneous performance of multiple treatments, but also cross contamination between samples must be avoided and high transformation efficiency must be ensured. Designing a device suitable for agrobacterium transient transfection of radix tetrastigme not only helps to improve the transformation efficiency, but also avoids cross infection between samples during transfection, which has important application value for the basic molecular biology research of radix tetrastigme.

[0004] A simple plant inverted vacuum infection device (authorized publication number CN219122161U) is designed to invert the plant, so that the aboveground part of the plant is located outside the device, and the underground part is placed in the folding flowerpot inside the device. The bottom of the device is equipped with an openable arc-shaped leaf, which can effectively block the soil by controlling the bottom opening, preventing the soil of the plant from falling off during the infection process, thereby ensuring the smooth progress of the infection experiment.

[0005] This vacuum infection device is specially designed with a soil blocking function to prevent soil contamination of the immersion liquid during the plant inversion infection process. Although similar devices exist in the market, they can usually only handle a single sample and are not suitable for handling plants such as radix tetrastigme with many branches and long branches. In addition, existing devices cannot meet the needs of simultaneous infection of multiple treatment strains. UTILITY MODEL CONTENTS

[0006] The utility model discloses a device for three-leaf ginseng agrobacterium transient infection, to solve the problem that only single sample can be handled by the existing infection device.

[0007] A device for three-leaf ginseng agrobacterium transient infection, comprising an infection assembly and a net bag, the infection assembly comprises a bracket, a tray, a container and an upper cover, a plurality of containers are placed on the tray, a plurality of upper covers correspondingly cover the plurality of containers, the upper cover is provided with a hole, the bracket is placed on the plurality of upper covers, the bracket is provided with an inner ring, the upper and lower ends of the net bag are provided with bag mouths, the bag mouth is provided with a tightening pull rope, three-leaf ginseng is planted in a flowerpot, the flowerpot is arranged in an inverted mode, the open end of the flowerpot is inserted into the net bag, and the two tightening pull ropes are tightened, the flowerpot is placed on the bracket, and a plurality of branches of the three-leaf ginseng are inserted into the plurality of containers through the hole from the bag mouth at the lower end of the net bag.

[0008] Further, the tray comprises a disc body, and the disc body is provided with a positioning cylinder in the center.

[0009] Further, the container is a transparent acrylic component, the circumferential contour of the container is in the shape of a fan ring, a plurality of containers are circumferentially arranged on the outer periphery of the positioning cylinder, and the inner periphery of the container is matched with the outer periphery of the positioning cylinder.

[0010] Further, the outer periphery of the container is provided with a strip-shaped protruding belt in the circumferential direction, the outer end of the upper cover is bent downward to form a hook-shaped edge, and when the upper cover covers the container, the hook-shaped edge is clamped on the lower end of the strip-shaped protruding belt.

[0011] Further, the number of the containers is four, and the central angle of the container is 90 degrees.

[0012] Further, the upper cover is in the shape of a fan ring, and the upper cover is matched with the upper opening of the container.

[0013] Further, the upper cover is circumferentially spliced by two fan ring units with the same central angle, and the hole is arranged on the joint of the two fan ring units.

[0014] Further, the bracket further comprises an outer supporting cylinder, the inner ring and the outer supporting cylinder are sleeved, the lower edge of the outer supporting cylinder is supported on the plurality of upper covers, and the inner ring and the outer supporting cylinder are connected through a plurality of supporting rods.

[0015] Further, the cross section of the inner ring is a circle with a diameter of 1 cm, the inner hole diameter of the inner ring is 5 cm, the outer diameter of the outer supporting cylinder is 25 cm, and the wall thickness of the outer supporting cylinder is 0.5 cm.

[0016] Further, the mesh aperture of the net bag is 0.2 cm.

[0017] Compared with the prior art, the utility model has the beneficial effects that:

[0018] The threeleaf ginseng leaf is transformed by using vacuum transient agrobacterium infection, which is the main method for molecular biology research of the threeleaf ginseng at present. A container is designed, which can simplify the steps of the threeleaf ginseng agrobacterium transient transfection by cooperating with the currently marketed vacuum container. The flowerpot is covered by a net bag, and the bag opening is tightened by tightening the pull rope, so that the soil can be prevented from falling into the container. Different strains of infection liquid are arranged in a plurality of containers, so that a plurality of treatments can be simultaneously infected, the infection process is shortened, and time is saved. The container is made of transparent acrylic material, so that the penetration of the leaf can be observed, and the infection effect can be tracked. Meanwhile, the acrylic material is acid-resistant, which is beneficial to the disinfection of the transgenic microorganism by using hypochlorite in the later period, avoids the spread of the transgenic microorganism, and increases the service life of the equipment. The utility model also specially considers the compatibility with the commonly used vacuum container on the market, and can well cooperate with the cylindrical vacuum container with a diameter of 40 cm, and is suitable for the vacuum infection experiment of the threeleaf ginseng agrobacterium. BRIEF DESCRIPTION OF DRAWINGS

[0019] Figure 1 is a structure diagram of the infection assembly;

[0020] Figure 2 is a structure diagram of the net bag;

[0021] Figure 3 is a top view of the utility model

[0022] Figure 4 is a structure diagram of the bracket;

[0023] Figure 5 is a structure diagram of the tray;

[0024] Figure 6 is a structure diagram of the container;

[0025] Figure 7 is a structure diagram of the upper cover;

[0026] Figure 8 is a use state diagram of the utility model for infecting threeleaf ginseng.

[0027] In the drawing, 1. tray, 11. disc body, 12. positioning cylinder, 2. container, 21. strip-shaped protruding belt, 3. upper cover, 31. fan ring unit, 32. hole, 33. hook-shaped edge, 4. bracket, 41. outer supporting cylinder, 42. inner ring, 43. supporting rod, 5. net bag, 51. bag opening, 52. tightening pull rope, 6. flowerpot. DETAILED DESCRIPTION

[0028] To make the objectives, technical solutions, and advantages of this utility model clearer, the present utility model is described below with reference to specific embodiments shown in the accompanying drawings. However, it should be understood that these descriptions are merely exemplary and not intended to limit the scope of the present utility model. Furthermore, descriptions of well-known structures and technologies are omitted in the following description to avoid unnecessarily obscuring the concept of the present utility model.

[0029] The connections mentioned in this utility model are divided into fixed connections and detachable connections. Fixed connections, also known as non-detachable connections, include but are not limited to conventional fixed connection methods such as folded connections, riveted connections, adhesive connections, and welded connections. Detachable connections include but are not limited to conventional disassembly methods such as bolt connections, snap-fit ​​connections, pin connections, and hinge connections. When a specific connection method is not explicitly defined, it is assumed that at least one existing connection method can be found to achieve this function, and those skilled in the art can choose according to their needs. For example, a welded connection can be chosen for a fixed connection, and a bolted connection can be chosen for a detachable connection.

[0030] The present invention will be further described in detail below with reference to the accompanying drawings. The following embodiments are explanations of the present invention, but the present invention is not limited to the following embodiments.

[0031] Example: Figures 1-8 As shown, an apparatus for instantaneous infection of Agrobacterium trifolium includes an infection component and a mesh bag 5. The infection component includes a bracket 4, a tray 1, containers 2, and a top cover 3. Several containers 2 are placed on the tray 1, and several top covers 3 are correspondingly sealed to several containers 2. The top covers 3 are provided with holes 32. The bracket 4 is placed on several top covers 3 and is provided with an inner ring 42. Both the upper and lower ends of the mesh bag 5 are provided with bag openings 51, and the bag openings 51 are provided with tightening ropes 52. Agrobacterium trifolium is planted in a flowerpot 6, which is set up upside down. The open end of the flowerpot 6 extends into the mesh bag 5, and the two tightening ropes 52 are tightened. The flowerpot 6 is placed on the bracket 4, and several branches of Agrobacterium trifolium pass through the bag openings 51 at the lower end of the mesh bag 5 and extend into several containers 2 through the holes 32.

[0032] Using vacuum-transferred Agrobacterium tumefaciens to transfect and transform *Trifolium repens* leaves is currently a major research method in the molecular biology of *Trifolium repens*. A container designed to work with commercially available vacuum containers can simplify the steps of instantaneous Agrobacterium tumefaciens transfection. A mesh bag 5 is used to cover the flowerpot 6, and the bag opening 51 is tightened by a drawstring 52 to prevent soil from falling into the container 2. Different strains of infection solutions can be placed in several containers 2, allowing for simultaneous infection of multiple treatments, shortening the infection process, and saving time.

[0033] In operation, the containers 2 are first filled with infection liquid containing Agrobacterium, several same-sized fan ring-shaped containers 2 are placed on the tray 1 and enclosed into a circular ring body, the flowerpot 6 is inverted on the bracket 4, then several branches of Radix Pteroxyprofini are immersed in different infection liquid, finally the whole device is placed in a vacuum column with a diameter of 40 cm, so that the infection process is started under vacuum condition.

[0034] The tray 1 comprises a disc body 11, and the disc body 11 is provided with a positioning cylinder 12 in the center. The disc body 11 is a hard plastic disc with a diameter of 30 cm, a thickness of 1 cm, and a cylinder with a diameter of 3.8 cm and a height of 19 cm in the center, which can be used as a positioning structure of the container 2.

[0035] The container 2 is a transparent acrylic component, and the circumferential profile of the container 2 is fan ring-shaped. The containers 2 are circumferentially arranged outside the outer periphery of the positioning cylinder 12, and the inner periphery of the container 2 matches the outer periphery of the positioning cylinder 12. The container 2 is made of transparent acrylic material, which can facilitate observation of the penetration of the leaves and tracking of the infection effect. At the same time, the acrylic material is acid-resistant, which is conducive to the disinfection of the transgenic microorganisms by using hypochlorite in the later stage, avoids the spread of the transgenic microorganisms, and increases the service life of the equipment.

[0036] The outer periphery of the container 2 is provided with a strip-shaped protruding belt 21 in the circumferential direction, and the outer end of the upper cover 3 is bent downward to form a hook-shaped edge 33. When the upper cover 3 covers the container 2, the hook-shaped edge 33 is clamped on the lower end of the strip-shaped protruding belt 21. The strip-shaped protruding belt 21 is used for fixing the upper cover 3.

[0037] The number of the containers 2 is four, and the central angle of the container 2 is 90°. The utility model also particularly considers the compatibility with the commercially available general vacuum container, which can well cooperate with the cylindrical vacuum container with a diameter of 40 cm, and is suitable for the vacuum infection experiment of Radix Pteroxyprofini Agrobacterium.

[0038] The upper cover 3 is fan ring-shaped, and the upper cover 3 matches the upper opening of the container 2.

[0039] The upper cover 3 is composed of two fan ring-shaped units 31 with the same central angle, and the hole 32 is arranged on the joint of the two fan ring-shaped units 31. The upper cover 3 composed of the two fan ring-shaped units 31 can be separated or combined, and the hole 32 is also formed by the half-circle notches on the two fan ring-shaped units 31, which is convenient for the branches of Radix Pteroxyprofini to pass through and does not damage the branches of Radix Pteroxyprofini.

[0040] The bracket 4 further comprises an outer supporting cylinder 41, and the inner circular ring 42 and the outer supporting cylinder 41 are sleeved. The lower edge of the outer supporting cylinder 41 is supported on the several upper covers 3, and the inner circular ring 42 and the outer supporting cylinder 41 are connected through the several supporting rods 43.

[0041] The outer supporting cylinder 41 plays the role of a supporting leg, and the inner ring 42 and the outer supporting cylinder 41 are connected through eight stainless steel supporting rods 43 arranged in the shape of a Chinese character'mi', the supporting rods 43 are used for supporting the flowerpot 6, the maximum load of the bracket 4 is 5 kg, and the branches of the Sanweiqing pass through the inner ring 42 and extend into the corresponding container 2 through the hole 32.

[0042] The section of the inner ring 42 is a circle with a diameter of 1 cm, the inner hole diameter of the inner ring 42 is 5 cm, the outer diameter of the outer supporting cylinder 41 is 25 cm, and the wall thickness of the outer supporting cylinder 41 is 0.5 cm. The inner ring 42 is free of sharp corners, and the Sanweiqing can be prevented from being damaged.

[0043] The mesh aperture of the net bag 5 is 0.2 cm.

[0044] The above examples are only illustrative of the present application and do not limit the protection scope thereof, and a person skilled in the art can also make changes to parts thereof, as long as the changes do not exceed the spirit and essence of the present application and are within the protection scope of the present application.

Claims

1. A device for transient infection by Agrobacterium trifoliatum, characterized in that: The device includes a dyeing assembly and a mesh bag (5). The dyeing assembly includes a bracket (4), a tray (1), containers (2), and a top cover (3). Several containers (2) are placed on the tray (1), and several top covers (3) are correspondingly sealed to several containers (2). The top covers (3) are provided with holes (32). The bracket (4) is placed on several top covers (3) and is provided with an inner ring (42). The mesh bag (5) is provided with bags at both the top and bottom ends. The bag opening (51) is equipped with a tightening rope (52). The three-leaf ginseng is planted in a flowerpot (6). The flowerpot (6) is placed upside down, and the open end of the flowerpot (6) extends into the net bag (5). The two tightening ropes (52) are tightened, and the flowerpot (6) is placed on the bracket (4). Several branches of the three-leaf ginseng pass through the bag opening (51) at the lower end of the net bag (5) and extend into several containers (2) through the holes (32).

2. The device for instantaneous infection of Agrobacterium trifolium according to claim 1, characterized in that: The tray (1) includes a disc body (11), and a positioning cylinder (12) is provided in the center of the disc body (11).

3. The device for instantaneous infection of Agrobacterium trifolium according to claim 2, characterized in that: The container (2) is a transparent acrylic component. The circumferential outline of the container (2) is a fan-shaped ring. Several containers (2) are arranged on the outer periphery of the positioning cylinder (12). The inner circumferential surface of the container (2) matches the outer periphery of the positioning cylinder (12).

4. The device for instantaneous infection of Agrobacterium trifolium according to claim 3, characterized in that: The top of the outer periphery of the container (2) is provided with a strip-shaped protrusion (21) along the circumferential direction. The outer end of the cover (3) is bent downward to form a hook-shaped edge (33). When the cover (3) seals the container (2), the hook-shaped edge (33) is stuck on the lower end of the strip-shaped protrusion (21).

5. The device for instantaneous infection of Agrobacterium trifolium according to claim 3, characterized in that: There are four containers (2), and the central angle of the container (2) is 90°.

6. The device for instantaneous infection of Agrobacterium trifolium according to claim 3, characterized in that: The top cover (3) is fan-shaped and fits the top opening of the container (2).

7. The device for instantaneous infection of Agrobacterium trifolium according to claim 6, characterized in that: The top cover (3) is formed by circumferentially joining two fan-shaped annular units (31) with the same central angle, and the hole (32) is opened on the joint of the two fan-shaped annular units (31).

8. The device for instantaneous infection of Agrobacterium trifolium according to claim 1, characterized in that: The bracket (4) also includes an outer support cylinder (41), an inner ring (42) and an outer support cylinder (41) are connected in an inner and outer sleeve, the lower edge of the outer support cylinder (41) is supported on several upper covers (3), and the inner ring (42) and the outer support cylinder (41) are connected by several support rods (43).

9. The device for instantaneous infection of Agrobacterium trifolium according to claim 8, characterized in that: The inner ring (42) has a cross-section with a diameter of 1cm, the inner hole diameter of the inner ring (42) is 5cm, the outer diameter of the outer support cylinder (41) is 25cm, and the wall thickness of the outer support cylinder (41) is 0.5cm.

10. The apparatus for transient infection of Agrobacterium trifolium according to any one of claims 1-9, characterized in that: The mesh size of the mesh bag (5) is 0.2cm.

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