RNA (Ribonucleic Acid) interference delivery device for small piercing-sucking insects

Abstract

The utility model provides an RNA (Ribonucleic Acid) interference delivery device for small piercing-sucking insects. The RNA interference delivery device comprises a test insect box, a thin film and a sealing cover, the test insect box is a cup-shaped container with a closed bottom surface and an open upper end; the thin film is a plastic film and covers the opening of the test insect box; the upper end of the sealing cover is closed, the bottom surface of the sealing cover is open, the sealing cover can be buckled at the opening of the test insect box, and an absorption pad is arranged in the sealing cover and is used for absorbing dsRNA; the absorption pad is in contact with the thin film; the sealing cover is made of a colored material or a detachable colored layer is arranged on the sealing cover. The device has the advantages of convenience in use, simplicity in operation, low cost and the like, solves the problems that time is wasted, the operation difficulty is high, precise instruments are needed, the cost is high and the like in the RNA interference delivery process, solves the problem that part of small-sized piercing-sucking insects cannot deliver dsRNA by using artificial feed, and is suitable for the RNA interference test of the small-sized piercing-sucking insects in a laboratory.

Description

Technical Field

[0001] The utility model relates to a test instrument, in particular to an RNA interference delivery device for small piercing-sucking insects. Background Art

[0002] The disclosure of this background information is intended to increase the understanding of the overall background of this patent, and should not necessarily be regarded as an admission or any form of suggestion that this information constitutes prior art that is already known to a person of ordinary skill in the art.

[0003] RNA interference (RNAi) is a phenomenon in which double-stranded RNA triggers mRNA degradation, thereby inhibiting the expression of specific target genes. It is specific and efficient, and can be used to screen target genes and study gene functions, and can also be used to develop RNAi pesticides for pest control.

[0004] The right delivery method is a key factor for the success of RNAi. Currently, RNAi delivery methods include feeding-mediated and cuticle penetration-mediated. Cuticle penetration-mediated RNAi delivery systems include injection and immersion. This delivery method can bypass the intestinal expression system, but the injection method is time-consuming and requires sophisticated instruments. The immersion method uses a large amount of double-stranded RNA and has high experimental costs. The feeding-mediated RNAi delivery system usually mixes double-stranded RNA with artificial insect feed. Insects that are not fed with artificial feed can also achieve RNAi effects by feeding transgenic plants expressing dsRNA, but the requirements for plant species are high and the amount of dsRNA delivered cannot be controlled.

[0005] Chinese patent CN 106942162 A discloses a device suitable for RNA interference experiments using the white-backed planthopper feeding method, in which dsRNA and artificial feed are fixed in the sealing layer of the white-backed planthopper feeding method RNA interference device through the setting of a double-layer membrane. However, this double-layer membrane setting is not suitable for direct feeding of dsRNA solution: first, the droplets will be spread out, resulting in uneven distribution and easy drying; second, the feeding behavior of the insects cannot be stimulated, both of which will affect the interference effect. Utility Model Content

[0006] In view of the current lack of convenient, efficient and low-cost RNA interference delivery devices for small piercing-sucking insects that cannot be fed with artificial feed, the utility model provides an RNA interference delivery device for small piercing-sucking insects, which has a simple structure, is easy to use and has low cost.

[0007] To achieve the above purpose, the utility model adopts the following technical solutions.

[0008] An RNA interference delivery device for small piercing-sucking insects, comprising a test insect box, a film and a cover;

[0009] The insect testing box is a cup-shaped container with a closed bottom and an open top;

[0010] The film is a plastic film, and the film is coated on the opening of the test insect box;

[0011] The upper end of the cover is closed, the bottom surface is open, and can be buckled at the opening of the insect testing box. An absorption pad is arranged inside the cover; and the absorption pad is in contact with the film.

[0012] In order to adapt to test insects in different habitats, the test insect box can also be provided with air holes for adjusting the humidity in the cup and increasing ventilation; at the same time, for test insects that are too small, in order to prevent them from escaping, a gauze is also provided on the air holes.

[0013] In order to increase the feeding amount of the test insects, an attracting layer is also provided on the film, and the attracting layer is on one side of the opening of the test insect box. The attracting layer contains a test insect attractant for attracting the test insects to feed.

[0014] In order to simulate the thickness of the epidermis of different plants, the thickness of the film can be adjusted. For example, food-use plastic wrap or laboratory sealing film can be used and stretched to the required thickness.

[0015] The absorption pad is used for absorbing the dsRNA solution, and its material can be natural or synthetic fiber, sponge, etc.

[0016] In order to facilitate opening of the cover, a lifting ear is provided on the edge of the cover.

[0017] In order to increase the feeding of the test insects, the cover is made of colored material, and the color of the test insects can be selected according to the type of test insects, such as green, yellow, brown or brown. In order to improve the universality of the device, the cover is colorless and transparent, and a detachable colored layer is also provided on its upper surface, such as a translucent or opaque color card is attached or placed on the cover. In order to facilitate the fixing of the colored layer, a plurality of fixing claws are also provided on the cover.

[0018] The size of the device can be adjusted according to the size and number of the test insects, and can be applied to agricultural and sanitary pests.

[0019] When the utility model is used, a number of test insects of a test quantity are placed in a test insect box, the film is disinfected with alcohol, then stretched to a suitable thickness, and an attractant solution of a suitable concentration is sprayed, and when the solvent evaporates, an attractant layer is formed, and then the attractant layer is sealed toward the inside of the test insect box, and then a dsRNA solution is quantitatively added to the absorption pad of the cover, and the cover is closed so that the absorption pad can contact the film without liquid flowing out; in order to increase feeding, a translucent color card is attached to the upper surface of the cover. According to the biological characteristics of the test insects, the device is placed under suitable temperature, humidity and light cycle conditions for 24h-96h to complete the RNA interference test.

[0020] The utility model has the following advantages:

[0021] The utility model provides a small-scale piercing-sucking insect RNA interference delivery device, which directly feeds the test insects by absorbing the dsRNA solution into the built-in absorption pad of the sealing cover, and the feeding and delivery of dsRNA can be completed without artificial feed; the absorption pad can accommodate a large amount of solution, is not easy to dry, and the liquid does not flow, which is easy to operate and can ensure that the test insects can eat the dsRNA solution during the entire experiment. The utility model lures insects to eat dsRNA solution from both the olfactory and visual aspects by applying an attractant on the side of the sealing film close to the test insects and setting the color of the sealing cover, thereby enhancing the RNA interference effect. In addition, the test insect box and the sealing cover with a built-in absorption pad of the utility model can be recycled after sterilization and RNAse-free treatment, which is more economical and environmentally friendly.

[0022] The utility model has the advantages of easy use, simple operation and low cost. It solves the problems of time-consuming, difficult operation, need for precision instruments and high cost in the RNA interference delivery process. It also solves the problem that some small piercing-sucking insects cannot use artificial feed to deliver dsRNA. The utility model is suitable for RNA interference experiments of small piercing-sucking insects in laboratories. BRIEF DESCRIPTION OF THE DRAWINGS

[0023] Figure 1 It is the structural schematic diagram of the test insect box and the film;

[0024] Figure 2 Schematic diagram of the structure of the cover and the absorbent pad in Examples 1 and 2;

[0025] Figure 3 It is a schematic diagram of the structure of the cover and the color card in Example 2;

[0026] Figure 4 is a schematic diagram of the structure of the cover and the absorbent pad in Example 3;

[0027] Figure 5 This is a schematic diagram of the structure of the cover and the color card in Example 3;

[0028] Figure 6 This is a schematic diagram of the structure of the RNA interference delivery device for small piercing-sucking insects in Example 3;

[0029] Among them, 1-test insect box, 2-film, 3-cover, 4-air vent, 5-absorption pad, 6-lifting ear, 7-color card, 8-fixing claw. DETAILED DESCRIPTION

[0030] The present invention will be further described below in conjunction with embodiments and drawings, but the present invention is not limited by the following embodiments.

[0031] Example 1

[0032] An RNA interference delivery device for small piercing-sucking insects, such as Figure 1 and Figure 2 As shown, it includes an insect test box 1, a film 2 and a cover 3;

[0033] The test insect box 1 is a cup-shaped container with a closed bottom and an open top. The test insect box 1 is also provided with a plurality of air holes 4 for adjusting the humidity in the cup and increasing ventilation.

[0034] The film 2 is a plastic sealing film. In order to simulate the thickness of the epidermis of different plants, the thickness of the film 2 can be adjusted. The film 2 covers the opening of the test insect box 1. In order to increase the feeding amount of the test insects and enhance the interference effect, an attractant layer is also provided on the film 2. The attractant layer contains a test insect attractant. The attractant layer is on one side of the opening of the test insect box.

[0035] The upper end of the cover 3 is closed and the bottom is open, and can be snapped into the opening of the test insect box 1. Seven sponge-made absorption pads 5 are arranged in an array inside the cover 3; the absorption pads 5 are slightly higher than the lower opening plane of the cover 3 and are in contact with the film 2; the cover 3 is translucent yellow; in order to facilitate the opening of the cover 3, a lifting ear 6 is provided on the edge of the cover 3.

[0036] The above device is used for RNA interference test of small piercing-sucking pests with body length less than 1.5 mm such as aphids. The opening diameter of the test insect box 1 is 30 mm and the volume is about 25 mL; the diameter of the cover 2 is slightly larger than the opening of the test insect box 1 and can be snapped on the opening of the test insect box 1, with a depth of about 8 mm; the absorption pad 5 is a rectangular high-density sponge with a length and width of 5 mm and a height of 8.5 mm.

[0037] Example 2

[0038] An RNA interference delivery device for small piercing-sucking insects, such as Figure 1-3 As shown, it includes an insect test box 1, a film 2 and a cover 3;

[0039] The test insect box 1 is a cup-shaped container with a closed bottom and an open top. The box body of the test insect box 1 is also provided with a plurality of air holes 4 for adjusting the humidity in the cup and increasing the ventilation volume. At the same time, in order to prevent the test insects from escaping, a gauze is also provided on the air holes 4.

[0040] The film 2 is a plastic sealing film. In order to simulate the thickness of the epidermis of different plants, the thickness of the film 2 can be adjusted. The film 2 covers the opening of the test insect box 1. In order to increase the feeding amount of the test insects and enhance the interference effect, an attractant layer is also provided on the film 2. The attractant layer contains a test insect attractant. The attractant layer is on one side of the opening of the test insect box 1.

[0041] The cover 3 is closed at the upper end and open at the bottom, is colorless and transparent, and can be snapped on the opening of the test insect box 1. Seven sponge absorption pads 5 are arranged in the cover 3; the absorption pads 5 are slightly higher than the lower opening plane of the cover and are in contact with the film 2; a translucent green plastic color card 7 is also attached to the cover 3; in order to facilitate the opening of the cover 3, a lifting ear 6 is arranged on the edge of the cover 3.

[0042] The above device is used for RNA interference test of small piercing-sucking pests with body length less than 1.5 mm such as whiteflies. According to the following, the opening diameter of the test insect box 1 is 30 mm, and the volume is about 25 mL; the caliber of the cover 3 is slightly larger than the opening of the test insect box 1 and can be snapped on the opening of the test insect box 1, with a depth of about 8 mm; the absorption pad 5 is a rectangular high-density sponge with a length and width of 8 mm and a height of 8.5 mm.

[0043] When the above device is used for RNAi interference test of small piercing-sucking pests with a body length of 2-3 mm such as leafhoppers, the opening diameter of the test insect box 1 is 50 mm and the volume is about 40 mL; the diameter of the cover is slightly larger than the opening of the test insect box 1 and can be snapped on the opening of the test insect box 1, with a depth of about 8 mm; the absorption pad 5 is a rectangular high-density sponge with a length and width of 8 mm and a height of 8.5 mm.

[0044] Example 3

[0045] An RNA interference delivery device for small piercing-sucking insects, such as Figure 1 and Figure 4-6 As shown, it includes an insect test box 1, a film 2 and a cover 3;

[0046] The test insect box 1 is a cup-shaped container with a closed bottom and an open top. The test insect box 1 is also provided with a plurality of air holes 4 for adjusting the humidity in the cup and increasing ventilation.

[0047] The film 2 is a plastic sealing film. In order to simulate the thickness of the epidermis of different plants, the thickness of the film 2 can be adjusted. The film 2 covers the opening of the test insect box 1. In order to increase the feeding amount of the test insects and enhance the interference effect, an attractant layer is also provided on the film 2. The attractant layer contains a test insect attractant. The attractant layer is on one side of the opening of the test insect box.

[0048] The cover 3 is closed at the upper end and open at the bottom, is colorless and transparent, and can be snapped into the opening of the test insect box 1. A sponge-made absorption pad 5 is arranged inside the cover 3; the absorption pad 5 is slightly higher than the lower opening plane of the cover 3 and is in contact with the film 2; four fixing claws 8 are arranged on the upper surface of the cover 3 for fixing a translucent plastic green color card 7; in order to facilitate the opening of the cover 3, a lifting ear 6 is arranged on the edge of the cover 3.

[0049] When the above device is used for RNA interference testing of small piercing-sucking pests with a body length of about 5 mm, such as plant hoppers, the opening diameter of the test insect box 1 is 70 mm and the volume is about 70 mL; the caliber of the cover 3 is slightly larger than the opening of the test insect box 1 and can be snapped on the opening of the test insect box 1, with a depth of about 8 mm; the absorption pad 5 is a rectangular high-density sponge with a length and width of 20 mm and a height of 8.5 mm.

[0050] When the above devices are used, the entire device is placed in a sterile environment for operation, 10-30 healthy test insects are selected and placed in the test insect box 1, and Parafilm ® The sealing film is stretched to a suitable thickness as a film 2, and the front and back sides are disinfected with 75% alcohol two or three times to evaporate the alcohol; then, an attractant solution of suitable concentration is sprayed, and when the solvent evaporates, an attractant layer is formed, and then the attractant layer is directed toward the inside of the test insect box 1, so that the film 2 is attached to the mouth of the test insect box 1 and tightened; then the dsRNA solution is quantitatively added to each absorption pad 5 of the cover 3 with a pipette, and the cover 3 is covered so that the absorption pad 5 can contact with the film 2 without liquid flowing out. According to the biological characteristics of the test insects, the device is placed under suitable temperature, humidity and light cycle conditions for 24h-96h to complete the RNA interference test. After the test is completed, the test insects are taken out, the film 2 is discarded, the device is cleaned and dried, sterilized and then used for standby.

[0051] The above is only an embodiment of the present application and is not intended to limit the present application. For those skilled in the art, the present application may have various changes and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present application should be included in the scope of the claims of the present application.

Claims

1. An RNA interference delivery device for small piercing-sucking insects, characterized in that: Including test insect box, film and cover; The insect testing box is a cup-shaped container with a closed bottom and an open top; The film is a plastic film, and the film is coated on the opening of the test insect box; The upper end of the cover is closed, the bottom surface is open, and can be snapped on the opening of the test insect box. An absorbent pad is arranged inside the cover; The absorbent pad is in contact with the film.

2. The RNA interference delivery device according to claim 1, characterized in that The insect testing box is also provided with air holes.

3. The RNA interference delivery device according to claim 2, characterized in that The ventilation hole is also provided with a gauze net.

4. The RNA interference delivery device according to claim 1, characterized in that The film is also provided with an attracting layer, and the attracting layer is located on one side of the opening of the insect testing box.

5. The RNA interference delivery device according to claim 1, characterized in that The material of the absorption pad is natural or synthetic fiber or sponge.

6. The RNA interference delivery device according to claim 1, characterized in that The sealing cover is made of colored material.

7. The RNA interference delivery device according to claim 1, characterized in that The sealing cover is colorless and transparent, and a detachable colored layer is provided on its upper surface.

8. The RNA interference delivery device according to claim 7, characterized in that: A plurality of fixing claws are also arranged on the sealing cover.

9. The RNA interference delivery device according to claim 1, characterized in that: The edge of the cover is provided with a lifting ear.

Images