Entomopathogenic nematode bioassay device

Abstract

The entomopathogenic nematode bioassay device comprises a bioassay plate 1, a bioassay cover 15, a gradienter 8, a sample plug 11 and a degumming brush 18, the bioassay plate 1 is provided with a bioassay area 2, an identification area 13 and a bioassay area number 3, and shading 17 is arranged at the bottom of the bioassay plate 1; the bioassay area 2 is provided with a cover groove 4 and a bioassay pool 6; the bioassay pool 6 is provided with ruler scales 7 and an indicating arrow 10; the bioassay pool 6 is provided with a height indicating line 14 and a sample adding hole 5; the gradienter 8 is provided with an indicating bubble 9; the sample plug 11 is provided with a plug head 12; finger anti-skid areas 16 are arranged on the two sides of the bioassay cover 15; the glue removing brush 18 is provided with a brush handle 19, brush handle anti-skid lines 20 and a brush head 21. The device has the advantages of being simple in structure, convenient to operate, high in working efficiency, wide in application range and the like, and has good practicability.

Description

TECHNICAL FIELD

[0001] The utility model belongs to the field of plant protection and related scientific and technological fields, and relates to a kind of entomopathogenic nematode bioassay device, mainly for pest biological control and related work. BACKGROUND

[0002] Entomopathogenic nematode refers to a kind of pathogenic nematode that parasitizes insects, which carries pathogenic bacteria in digestive tract. When it invades into insect host from digestive tract or body wall, symbiotic bacteria are released from the body of nematode, proliferate in insect blood and eventually cause host insect to die of septicemia. Entomopathogenic nematode is a natural enemy of pests and has become an important group in biological pest control, including Steinernema and Heterorhabditis, which have attracted increasing attention in biological pest control. Entomopathogenic nematode has the ability to actively search for host insects and can be recycled in natural environment. It has been successfully applied to control a variety of agricultural and forestry pests such as scarab, corn rootworm, agrotis segetum, peach fruitworm and spodoptera frugiperda.

[0003] Nematode has a highly developed chemical sensing system, and the head receptor is mainly used to sense temperature, chemical substances and other stimuli. In recent years, the chemical communication mechanism among entomopathogenic nematode, host insect and host plant has attracted attention. For example, Li Chunjie et al. (2019) found that the chemical signal released by leek roots can attract entomopathogenic nematodes to the roots over a long distance, while the chemical signal released by host insect leekworm can attract entomopathogenic nematodes as a short distance signal, thereby realizing the positioning, recognition and invasion of entomopathogenic nematodes to host insects. Therefore, it is of great significance to study the interaction among entomopathogenic nematode, host insect and host plant, screen substances with attractive activity to entomopathogenic nematode, and attract entomopathogenic nematode to host insect at long and short distances, so as to improve the infection rate and control efficiency of entomopathogenic nematode.

[0004] At present, the behavior activity of nematode is mainly determined by agar plate method, but there are a series of problems such as nematode wandering on agar plate, long wandering time, difficulty in counting, large amount of plate, etc., which greatly affect the detection efficiency and accuracy. Therefore, it is very important to continuously innovate and improve related equipment and devices in order to promote the behavior selection of nematode and improve the reaction efficiency of nematode to chemical substances.

[0005] According to the actual needs of nematode behavior test, a simple and practical entomopathogenic nematode bioassay device is developed, which has the advantages of simple structure, convenient operation, high working efficiency, accurate determination result and wide application range, etc., and has good practicability, which has important practical significance in pest biological control and related work. SUMMARY

[0006] The utility model provides a kind of entomopathogenic nematode bioassay device for the prior art deficiency, it includes: bioassay plate, bioassay cover, level, sample plug, glue removing brush;Bioassay cover is clamped in the cover groove around bioassay area;Sample plug is used to plug in sample addition hole when making glue bed;Level is embedded on bioassay plate.

[0007] The bioassay plate of the entomopathogenic nematode bioassay device has a bioassay area, an identification area, and a bioassay area number. The bioassay area has a cover groove around its perimeter, a bioassay pool in the middle, scale markings on both sides of the upper side, and three indicator arrows in the middle of the lower side. The bioassay pool has height indicator lines on both sides and one sample addition hole at each end.

[0008] The bioassay plate of the entomopathogenic nematode bioassay device has a bottom pattern.

[0009] The level of the entomopathogenic nematode bioassay device has an indicator bubble.

[0010] The sample plug of the entomopathogenic nematode bioassay device has a plug head.

[0011] The bioassay cover of the entomopathogenic nematode bioassay device has finger anti-slip areas on both sides.

[0012] The glue removing brush of the entomopathogenic nematode bioassay device has a brush handle and a brush head, and the brush handle has brush handle anti-slip lines.

[0013] The utility model has the advantages of simple structure, easy operation, high work efficiency, wide application range, and good practicality.

[0014] The utility model will be described in detail in combination with the drawings and examples. BRIEF DESCRIPTION OF DRAWINGS

[0015] Figure 1 It is a top view of the utility model.

[0016] Figure 2 It is a bioassay area structure schematic view of the utility model.

[0017] Figure 3 It is a bioassay cover structure schematic view of the utility model.

[0018] Figure 4 It is a bottom plate structure schematic view of the utility model.

[0019] Figure 5 It is a glue removing brush structure schematic view of the utility model.

[0020] In the diagram, the components are: 1. Bioassay plate; 2. Bioassay area; 3. Bioassay area number; 4. Cover groove; 5. Sample filling hole; 6. Bioassay cell; 7. Scale markings; 8. Level; 9. Indicating bubble; 10. Indicating arrow; 11. Sample plug; 12. Plug head; 13. Marking area; 14. Height indicator line; 15. Bioassay cover; 16. Anti-slip area for fingers; 17. Background texture; 18. Adhesive removal brush; 19. Brush handle; 20. Anti-slip texture on brush handle; 21. Brush head. Detailed Implementation

[0021] like Figures 1 to 5 As shown, a bioassay device for entomopathogenic nematodes includes: a bioassay plate 1, a bioassay cover 15, a level 8, a sample plug 11, and a descaling brush 18.

[0022] The bioassay plate 1 of the bioassay device for entomopathogenic nematodes has six bioassay zones 2 and a marking area 13 at the bottom; different bioassay zones 2 correspond to different bioassay zone numbers 3; the bioassay zone 2 has a cover groove 4 around its perimeter and a bioassay pool 6 in the middle; the bioassay pool 6 has scale markings 7 on the upper two sides and three indicator arrows 10 (left, center, and right) in the lower middle part; the bioassay pool 6 has height indicator lines 14 on both sides inside and a sample feeding hole 5 at each end.

[0023] The bioassay plate 1 of the entomopathogenic nematode bioassay device has a bottom texture 17.

[0024] The level 8 of the entomopathogenic nematode biodetection device has an indicator bubble 9.

[0025] The sample plug 11 of the entomopathogenic nematode bioassay device has a plug head 12.

[0026] The biodetection cover 15 of the entomopathogenic nematode biodetection device has anti-slip finger areas 16 on both sides.

[0027] The adhesive removal brush 18 of the insect pathogen nematode biodetection device has a brush handle 19 and a brush head 21. The brush handle 21 has anti-slip texture 20.

[0028] The utility model discloses a use method: every living test board 1 has six living test areas 2, and every living test area 2 corresponds to a living test area number 3, can carry out six times repeated test simultaneously;Before using, in the identification area 13 of living test board 1, mark relevant determination information (such as: name, date etc.) with a marker pen, through the sample plug 11 of plug head 12 is inserted into the sample adding hole 5 of both ends of living test pool 6, and is placed on the horizontal operation platform, through whether the indicating bubble 9 in level gauge 8 is in the middle judges whether living test board 1 is placed horizontally, avoids the agar bed made in living test pool 6 not horizontal, and the bottom of living test board 1 has the bottom line 17 and avoids the adsorption of the bottom of living test board 1 and test bench surface;The upper two sides of living test pool 6 have scale graduation 7, and the lower middle part has left, middle and right three indicating arrows 10, and the area between the left and right two indicating arrows 10 is the nematode suspension adding area, and the middle indicating arrow 10 indicates that the nematode suspension is added to the area from the middle;The total length of living test pool 6 is 5cm, and the distance between the left and right two indicating arrows 10 is 1cm, and the depth of living test pool 6 is 5mm, and the width of the non-nematode suspension adding area is 2mm;Utilize agar powder or agarose to make agar gel (according to the need, take the quantitative agar powder or agarose, and add the quantitative deionized water, heat and dissolve), and the agar hot solution is added to living test pool 6 to the height indicating line 14 by using the pipette gun, after cooling and solidifying, gently pull out the sample plug 11 in the sample adding hole 5 of both ends of living test pool 6, set up the treatment end and the control end, add a certain amount (such as: 10ul) of the agent to be tested to the treatment end, and a certain amount (such as: 10ul) of the control solution (such as: deionized water) can be added to the control end;The area between the two indicating arrows 10 is the nematode suspension drop adding area, and a certain volume (such as: 10ul) of the nematode suspension containing a certain amount of nematode is added to the nematode suspension adding area (the area between the left and right two indicating arrows 10) in the middle of living test pool 6 lower side middle indicating arrow 10;After being placed in the living test area 2 periphery cover groove 4 by pinching living test cover 15 through the finger anti -skid area 16 of both sides of living test cover 15, place in the constant temperature incubator, cultivate 2h under 26 DEG C and light -proof, take out and observe under the stereoscopic microscope, according to the scale graduation 7 of the upper two sides of living test area 2, record the position of insect pathogenic nematode, and statistics nematode quantity etc.;According to the number of nematodes in the treatment end and the total number of nematodes, calculate the attraction rate (%), and the calculation formula is attraction rate (%)=(the number of nematodes in the treatment end / the total number of nematodes)×100, wherein the total number of nematodes is the number of observed nematodes in living test pool 6;After completing living test, remove living test cover 15, and clean living test board 1, and the cleaning can use the brush 18 to remove the waste glue in living test pool 6, and the specific operation is: the brush head 21 is inserted into living test pool 6 by pinching the brush handle 19 through the brush handle anti -skid line 20 of finger, and the waste glue is removed;After cleaning the living test device, store for standby.

[0029] The utility model discloses a use method: every living test board 1 has six living test areas 2, and every living test area 2 corresponds to a living test area number 3, can carry out six times repeated test simultaneously;Before using, in the identification area 13 of living test board 1, mark relevant determination information (such as: name, date etc.) with a marker pen, through the sample plug 11 of plug head 12 is inserted into the sample adding hole 5 of both ends of living test pool 6, and is placed on the horizontal operation platform, through whether the indicating bubble 9 in level gauge 8 is in the middle judges whether living test board 1 is placed horizontally, avoids the agar bed made in living test pool 6 not horizontal, and the bottom of living test board 1 has the bottom line 17 and avoids the adsorption of the bottom of living test board 1 and test bench surface;The upper two sides of living test pool 6 have scale graduation 7, and the lower middle part has left, middle and right three indicating arrows 10, and the area between the left and right two indicating arrows 10 is the nematode suspension adding area, and the middle indicating arrow 10 indicates that the nematode suspension is added to the area from the middle;The total length of living test pool 6 is 5cm, and the distance between the left and right two indicating arrows 10 is 1cm, and the depth of living test pool 6 is 5mm, and the width of the non-nematode suspension adding area is 2mm;Utilize agar powder or agarose to make agar gel (according to the need, take the quantitative agar powder or agarose, and add the quantitative deionized water, heat and dissolve), and the agar hot solution is added to living test pool 6 to the height indicating line 14 by using the pipette gun, after cooling and solidifying, gently pull out the sample plug 11 in the sample adding hole 5 of both ends of living test pool 6, set up the treatment end and the control end, add a certain amount (such as: 10ul) of the agent to be tested to the treatment end, and a certain amount (such as: 10ul) of the control solution (such as: deionized water) can be added to the control end;The area between the two indicating arrows 10 is the nematode suspension drop adding area, and a certain volume (such as: 10ul) of the nematode suspension containing a certain amount of nematode is added to the nematode suspension adding area (the area between the left and right two indicating arrows 10) in the middle of living test pool 6 lower side middle indicating arrow 10;After being placed in the living test area 2 periphery cover groove 4 by pinching living test cover 15 through the finger anti -skid area 16 of both sides of living test cover 15, place in the constant temperature incubator, cultivate 2h under 26 DEG C and light -proof, take out and observe under the stereoscopic microscope, according to the scale graduation 7 of the upper two sides of living test area 2, record the position of insect pathogenic nematode, and statistics nematode quantity etc.

Claims

1. An apparatus for the bioassay of entomopathogenic nematodes, comprising: The plate (1), cover (15), level (8), sample plug (11), and glue removal brush (18) are characterized in that the plate (1) has six test zones (2), a lower identification zone (13), and a bottom with a bottom line (17). Different test zones (2) correspond to different test zone numbers (3). The periphery of the test zone (2) has a cover groove (4), and the middle has a test pool (6). The upper two sides of the test pool (6) have scale markings (7), the lower middle has left, middle, and right indication arrows (10), the inside has height indication lines (14), and each end has a sample hole (5). The test cover (15) is clamped in the cover groove (4) around the test zone (2). The sample plug (11) has a plug head (12) that can plug the sample hole (5). The level (8) is embedded on the plate (1) and has an indication bubble (9) in the middle.

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