CN116106478B - A bioassay device and method for detecting the sensitivity of the American white moth to Cry1Ab toxin - Google Patents
A bioassay device and method for detecting the sensitivity of the American white moth to Cry1Ab toxin Download PDFInfo
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- CN116106478B CN116106478B CN202211463805.0A CN202211463805A CN116106478B CN 116106478 B CN116106478 B CN 116106478B CN 202211463805 A CN202211463805 A CN 202211463805A CN 116106478 B CN116106478 B CN 116106478B
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- 241001531327 Hyphantria cunea Species 0.000 title claims abstract description 33
- 230000035945 sensitivity Effects 0.000 title claims abstract description 25
- 239000003053 toxin Substances 0.000 title claims abstract description 25
- 231100000765 toxin Toxicity 0.000 title claims abstract description 25
- 238000004166 bioassay Methods 0.000 title claims abstract description 22
- 238000000034 method Methods 0.000 title claims abstract description 7
- 238000012360 testing method Methods 0.000 claims abstract description 74
- 239000003153 chemical reaction reagent Substances 0.000 claims abstract description 68
- 238000007789 sealing Methods 0.000 claims abstract description 32
- 238000005192 partition Methods 0.000 claims description 32
- 108700012359 toxins Proteins 0.000 claims description 22
- 125000006850 spacer group Chemical group 0.000 claims description 6
- 230000005540 biological transmission Effects 0.000 claims description 5
- 230000002550 fecal effect Effects 0.000 claims description 3
- 230000004083 survival effect Effects 0.000 claims description 3
- 238000005259 measurement Methods 0.000 abstract description 3
- 238000005457 optimization Methods 0.000 description 5
- 101150063057 CRYAB gene Proteins 0.000 description 3
- 238000006073 displacement reaction Methods 0.000 description 3
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000193388 Bacillus thuringiensis Species 0.000 description 1
- 101150102464 Cry1 gene Proteins 0.000 description 1
- 241000282414 Homo sapiens Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 229930003270 Vitamin B Natural products 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 235000010323 ascorbic acid Nutrition 0.000 description 1
- 229960005070 ascorbic acid Drugs 0.000 description 1
- 239000011668 ascorbic acid Substances 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 230000001066 destructive effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- BEFDCLMNVWHSGT-UHFFFAOYSA-N ethenylcyclopentane Chemical compound C=CC1CCCC1 BEFDCLMNVWHSGT-UHFFFAOYSA-N 0.000 description 1
- 230000002431 foraging effect Effects 0.000 description 1
- 230000000749 insecticidal effect Effects 0.000 description 1
- 210000001161 mammalian embryo Anatomy 0.000 description 1
- 235000010270 methyl p-hydroxybenzoate Nutrition 0.000 description 1
- 239000004292 methyl p-hydroxybenzoate Substances 0.000 description 1
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 1
- 229960002216 methylparaben Drugs 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 235000010199 sorbic acid Nutrition 0.000 description 1
- 239000004334 sorbic acid Substances 0.000 description 1
- 229940075582 sorbic acid Drugs 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 238000009423 ventilation Methods 0.000 description 1
- 235000019156 vitamin B Nutrition 0.000 description 1
- 239000011720 vitamin B Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/30—Rearing or breeding invertebrates
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
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- Zoology (AREA)
- Animal Behavior & Ethology (AREA)
- Food Science & Technology (AREA)
- Medicinal Chemistry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Analytical Chemistry (AREA)
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- General Health & Medical Sciences (AREA)
- General Physics & Mathematics (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
Abstract
The invention relates to a biological assay device and a biological assay method for detecting sensitivity of fall webworm to Cry1Ab toxin, and relates to the technical field of fall webworm control, wherein the biological assay device comprises a test box, and four test cavities with upward openings are equally divided in the test box; the test device comprises a test box, a reagent tube group, a sealing cover assembly and a driving assembly, wherein the sealing cover assembly is arranged on the test box, the reagent tube group is clamped on the sealing cover assembly, the driving assembly is arranged right above the reagent tube group and comprises a rotating shaft rod and a pushing piece arranged on the rotating shaft rod, the pushing piece is in sliding fit with the tube wall of each reagent tube of the reagent tube group under the pushing of the rotating shaft rod, the sealing cover assembly comprises a swinging frame, a sealing cover piece and a locking seat, a rotating piece is arranged in the locking seat, and the locking seat is rotated to enable one end of the rotating shaft rod to be connected with the rotating piece while the locking seat drives the swinging frame to rotate so as to control the opening of the sealing cover piece. The invention can realize batch measurement of sensitivity of fall webworm to Cry1Ab toxin, and has strong testing flexibility and high efficiency.
Description
Technical Field
The invention belongs to the technical field of fall webworm control, and particularly relates to a biological assay device and a biological assay method for detecting sensitivity of fall webworm to Cry1Ab toxin.
Background
The fall webworm is a destructive pest and is listed as an international quarantine object, the development of some biological pesticides can effectively control the occurrence of fall webworm disasters at present, for example, delta-endotoxin generated by bacillus thuringiensis Bacillus thurigensis (Bt) has high insecticidal activity on the fall webworm, is safe to natural enemies and human beings and animals, and based on the thought, the damage of the fall webworm can be better controlled if Bt transgenic crops are developed;
Along with large-scale planting of Bt gene-transferred crops, the fall webworm has resistance to Bt proteins, so that the fall webworm needs to detect Cry1Ab toxin sensitivity, and a theoretical basis is provided for comprehensive treatment of the fall webworm.
Disclosure of Invention
The present invention aims to solve the above problems by providing a bioassay device and a bioassay method for detecting sensitivity of fall webworm to Cry1Ab toxin, so as to realize efficient and batched detection.
The invention realizes the above purpose through the following technical scheme:
The invention provides a biological assay device for detecting sensitivity of fall webworm to Cry1Ab toxin, which comprises a test box, wherein four test cavities with upward openings are formed in the inner part of the test box;
The test device comprises a sealing cover assembly and a reagent tube group, wherein the sealing cover assembly and the reagent tube group are arranged on the test box, and the number of the reagent tubes of the reagent tube group is consistent with that of the test cavities;
The driving assembly is arranged right above the reagent tube group and comprises a rotating shaft rod and a pushing piece arranged on the rotating shaft rod, and the pushing piece is in sliding fit with the tube wall of each reagent tube of the reagent tube group under the pushing of the rotating shaft rod;
The sealing cover assembly comprises a swing frame and a sealing cover piece, the outer side of the swing frame is connected with a locking seat, a rotating piece is arranged in the locking seat, and the locking seat is rotated to enable one end of a rotating shaft rod to be connected with the rotating piece while driving the swing frame to rotate so as to control opening of the sealing cover piece.
As a further optimization scheme of the invention, the structure of the single reagent tube in the reagent tube group comprises an outer tube body, a conical tube arranged at the lower end of the outer tube body and a jet nozzle arranged at the lower end of the conical tube.
As a further optimization scheme of the invention, the pushing piece comprises a transverse frame, a nut seat arranged on the transverse frame and a threaded sleeve sleeved on a rotating shaft rod and in threaded fit with the nut seat, pushing rods corresponding to the number of the reagent tubes of the reagent tube group are arranged on the lower end face of the transverse frame, and the upper end of the rotating shaft rod is driven to rotate by a connecting driving motor.
As a further optimization scheme of the invention, the locking seat comprises an axle seat, an external thread part is arranged at one end of the rotating shaft rod extending into the axle seat, an internal thread part matched with the external thread part is arranged on the inner side wall of the axle seat, the rotating member comprises a ball seat arranged in the axle seat and a ball block movably arranged in the ball seat, an inserting rod inserted with the ball block is arranged at the lower end of the rotating shaft rod, a shaft sleeve is arranged outside the rotating shaft rod, and the meshing transmission distance between the external thread part of the rotating shaft rod and the internal thread part is equal to the inserting distance between the inserting rod and the ball block.
As a further optimization scheme of the invention, the sealing cover part comprises a top cover, through holes corresponding to the positions of the test cavities are formed in the top cover, four partition frames are uniformly distributed on the top cover, a partition cover is matched in a rotating mode inside the partition frames, swing rods connected with the partition cover are arranged on the outer sides of the swing frames, corresponding to the partition frames, of the partition frames, and partition nets corresponding to the through holes are arranged on the partition covers.
As a further optimized scheme of the invention, the partition frame comprises a frame plate, a rotating shaft is rotatably matched in the frame plate through the arrangement of a torsion spring, and one end of the partition cover is connected with the rotating shaft and is wound on the rotating shaft.
As a further optimization scheme of the invention, the lower end of the test box is embedded with the excrement accumulating plate, and each test cavity of the test box is internally provided with a foraging plate.
The invention also provides a method for detecting the sensitivity of fall webworm to CryAb toxin by using the bioassay device as described in any one of the above steps,
Step one, putting the white moth larvae grouped according to the same quantity into each test cavity of a test box correspondingly, and keeping the white moth larvae fed under the same feeding environment;
Step two, preparing CryAb feeds with different concentrations, taking the same amount of feeds, respectively placing the feeds into each reagent tube of the reagent tube group, extending one end of a rotating shaft rod into a locking seat to enable a pushing piece to correspondingly extend into the tube orifice of each reagent tube, rotating the locking seat to enable one end of the rotating shaft rod to be connected with the rotating piece, and simultaneously enabling the locking seat to drive a swing frame to rotate so as to control the opening of a sealing cover piece, so that the opening of each test cavity is in an open state;
thirdly, driving the pushing part to push CryAb feed in each reagent tube to be fed from the opening of each test cavity after rotating the rotating shaft rod, feeding white moth larvae in each test cavity, stopping rotating the rotating shaft rod, rotating the locking seat to disconnect one end of the rotating shaft rod from the rotating part, and simultaneously driving the swinging frame to rotate by the locking seat to control the closing of the sealing cover part, so that the opening of each test cavity is in a closed state;
And step four, setting an observation period, observing the survival rate of the white moth larvae after feeding, and judging the sensitivity of the fall webworms to Cry1Ab toxins.
The invention has the beneficial effects that:
(1) According to the invention, the pushing rod is driven by the rotating shaft rod, and meanwhile, the artificial reagent mixed with the test reagent in each reagent tube of the test tube group is pushed into each test cavity of the test box, so that the operation is simple and quick, batch operation can be realized, and the efficiency is high;
(2) The device has strong structural flexibility, and when the locking seat is rotated to enable one end of the rotating shaft rod to be connected with the rotating piece, the locking seat drives the swinging frame to rotate so as to control the opening of the sealing cover piece, so that the pushing of the reagent into the testing cavity is not hindered, and the sealing performance of the testing cavity can be ensured after the reagent pushing is finished.
(3) The bioassay method provided by the invention not only can realize batch measurement of Cry1Ab toxin sensitivity of the fall webworm at the same concentration or concentration, but also can realize batch measurement of Cry1Ab toxin sensitivity of the fall webworm at different concentrations or different dosages, and has strong test flexibility and high efficiency.
Drawings
FIG. 1 is a schematic view of the overall structure provided by the present invention;
FIG. 2 is a schematic diagram illustrating the connection between a locking seat and a driving assembly according to the present invention;
FIG. 3 is a schematic view of a capping assembly according to the present invention;
FIG. 4 is a perspective view of a cartridge according to the present invention;
FIG. 5 is a schematic view of the internal structure of the bulkhead according to the present invention;
The device comprises a test box 1, a cross partition plate 2, a reagent tube group 31, an outer tube body 32, a conical tube 33, a jet nozzle 4, a driving component 41, a transverse frame 42, a pushing rod 43, a rotating shaft rod 44, a nut seat 45, a driving motor 46, a screw sleeve 47, a shaft sleeve 48, an external thread part 49, a plug rod 5, a sealing component 51, a top cover 52, a swinging frame 53, a swinging rod 53, a partition frame 54, a 541, a frame plate 542, a rotating shaft 543, a torsion spring 55, a partition cover 56, a partition net 6, a locking seat 61, an axle seat 62, a ball seat 63, a ball block 64, an internal thread part 7, a excrement deposition plate 8 and a food-seeking plate.
Detailed Description
The present application will be described in further detail with reference to the accompanying drawings, wherein it is to be understood that the following detailed description is for the purpose of further illustrating the application only and is not to be construed as limiting the scope of the application, as various insubstantial modifications and adaptations of the application to those skilled in the art can be made in light of the foregoing disclosure.
Example 1
As shown in fig. 1-3, the bioassay device for detecting sensitivity of fall webworm to Cry1Ab toxin provided in this embodiment includes a test box 1, and four test chambers with upward openings are equally divided in the test box 1 by a cross partition board 2;
The test device comprises a sealing cover assembly 5 and a reagent tube group 3, wherein the sealing cover assembly 5 and the reagent tube group 3 are arranged on the test box 1 and are clamped on the sealing cover assembly 5, and the number of the reagent tubes of the reagent tube group 3 is consistent with that of the test cavities;
The driving component 4 is arranged right above the reagent tube group 3, the driving component 4 comprises a rotating shaft rod 43 and a pushing piece arranged on the rotating shaft rod 43, and the pushing piece is in sliding fit with the tube wall of each reagent tube of the reagent tube group 3 under the pushing of the rotating shaft rod 43;
As shown in fig. 4-5, the cover assembly 5 includes a swing frame 52 and a cover member, a locking seat 6 is connected to the outer side of the swing frame 52, a rotating member is disposed in the locking seat 6, and the locking seat 6 is rotated to enable one end of the rotating shaft 43 to be connected with the rotating member, and the locking seat 6 drives the swing frame 52 to rotate to control opening of the cover member.
Specifically, the structure of a single reagent tube in the reagent tube set 3 includes an outer tube body 31, a conical tube 32 arranged at the lower end of the outer tube body 31, and a jet nozzle 33 arranged at the lower end of the conical tube 32, wherein the reagent tube set 3 formed by a plurality of reagent tubes can be matched to measure the sensitivity of fall webworm to Cry1Ab toxin under different concentrations or different dosages, and the arrangement of the jet nozzle 33 is beneficial to the rapid input of artificial feed mixed with test reagents into a test cavity.
As shown in fig. 1, the pushing member includes a transverse frame 41, a nut seat 44 disposed on the transverse frame 41, and a threaded sleeve 46 sleeved on the rotating shaft 43 and in threaded engagement with the nut seat 44, the lower end surface of the transverse frame 41 is provided with pushing rods 42 corresponding to the number of reagent tubes of the reagent tube set 3, the upper end of the rotating shaft 43 is driven to rotate by a driving motor 45, and when the pushing member is specifically used, the driving motor 45 drives the rotating shaft 43 to rotate, and then the threaded sleeve 46 outside the rotating shaft rotates together with the rotating shaft 43, so that the nut seat 44 on the threaded sleeve 46 generates vertical displacement, and the nut seat 44 is mounted on the transverse frame 41, thereby the transverse frame 41 and the pushing rods 42 on the transverse frame 41 can also generate vertical displacement, and the pushing rods 42 and the tube walls of the reagent tubes are in a state of abutting sliding fit, so that the test reagent in the reagent tubes can be pushed out when the pushing rods 42 generate vertical displacement.
As shown in fig. 2, the locking seat 6 includes an axle seat 61, an external threaded portion 48 is disposed at an end of the rotating shaft 43 extending into the axle seat 61, an internal threaded portion 64 matching with the external threaded portion 48 is disposed on an inner side wall of the axle seat 61, the rotating member includes a ball seat 62 disposed in the axle seat 61 and a ball block 63 movably disposed in the ball seat 62, a plugging rod 49 plugging with the ball block 63 is disposed at a lower end of the rotating shaft 43, a shaft sleeve 47 is disposed outside the rotating shaft 43, and a meshing transmission distance between the external threaded portion 48 and the internal threaded portion 64 of the rotating shaft 43 is equal to a plugging distance between the plugging rod 49 and the ball block 63;
The sealing cover part comprises a top cover 51, through holes corresponding to the positions of the test cavities are formed in the top cover 51, four partition frames 54 are uniformly distributed on the top cover 51, a partition cover 55 is rotatably matched in the partition frames 54, a swing rod 53 connected with the partition cover 55 is arranged on the outer side of the swing frame 52 corresponding to the outer side of the partition frames 54, and a partition net 56 corresponding to the through holes is arranged on the partition cover 55;
When the test reagent tube is specifically used, the swing frame 52 on the outer side of the swing frame 52 is driven to rotate during rotation of the shaft seat 61, so that the swing rod 53 on the outer side of the swing frame 52 sends the separation cover 55 into the separation frame 54, the opening of the top cover 51 corresponding to each test cavity is opened, on the other hand, when the shaft seat 61 rotates, the inner threaded part 64 on the inner side of the swing frame is meshed with the outer threaded part 48 on the lower end of the swing shaft 43, so that the swing shaft 43 is displaced and extends into the shaft seat 61, after the inner threaded part 64 is meshed with the outer threaded part 48, the inserting rod 49 on the lower end of the swing shaft 43 is just inserted into the ball block 63, the hole for inserting the inserting rod 49 is formed in the ball block 63, meanwhile, the shaft sleeve 47 arranged outside the swing shaft 43 is movably inserted into the upper end opening of the shaft seat 61, and the ball block 63 inserted into the lower end of the swing shaft 43 is in a rotating fit with the ball seat 62, so that the transmission structure of the shaft seat 61 and the sealing cover is not influenced, and the test reagent tube is pushed into each test cavity by the rotating and driving the pushing member by the rotating shaft 43;
After the test reagent is pushed, the rotation of the rotating shaft lever 43 is stopped, the rotating shaft lever 43 is lifted up to separate the inserting rod 49 from the ball block 63, the external thread part 48 of the rotating shaft lever 43 is contacted with the internal thread part 64, the swinging rod 53 of the swinging frame 52 at the outer side of the rotating shaft lever 43 is pulled out of the partition frame 54 by the rotating shaft seat 61, the partition net 56 on the partition frame 55 is only left for ventilation of the test cavity, the external thread part 48 and the internal thread part 64 generate meshing transmission, the rotating shaft lever 43 and the pushing piece can be separated from the structural connection with the locking seat 6 and the reagent tube set 3 respectively, and then the reagent tube set 3 is detached to be convenient for subsequent cleaning.
As shown in fig. 5, the spacer frame 54 includes a frame plate 541, a rotation shaft 542 is rotatably fitted in the frame plate 541 by providing a torsion spring 543, one end of the spacer cover 55 is connected to the rotation shaft 542 and wound around the rotation shaft 542, and the spacer cover 55 is made to be more smoothly retracted into the frame plate 541 by providing the torsion spring 543 with an automatic resetting property of the torsion spring 543, so that the pushing of the test reagent is not affected by being stuck outside the spacer frame 54.
As shown in fig. 3, the lower end of the test box 1 is embedded with a fecal accumulating plate 7 for receiving fecal discharged by the white moth larvae, and each test cavity of the test box 1 is provided with a feeding plate 8 for receiving artificial feed mixed with a test reagent for feeding the white moth larvae.
Example 2
This embodiment provides a method for detecting sensitivity of fall webworm to Cry1Ab toxin by using the bioassay device as described in any one of the above, comprising the steps of,
Firstly, putting the white moth larvae grouped according to the same quantity into each test cavity of the test box 1 correspondingly, and keeping the white moth larvae fed under the same feeding environment;
Step two, preparing artificial feed containing Cry1Ab with different concentrations, taking the same amount of the artificial feed and placing the artificial feed into each reagent tube of the reagent tube group 3, extending one end of a rotating shaft rod 43 into a locking seat 6 to enable a pushing piece to correspondingly extend into the tube orifice of each reagent tube, rotating the locking seat 6 to enable one end of the rotating shaft rod 43 to be connected with the rotating piece, and simultaneously enabling the locking seat 6 to drive a swinging frame 52 to rotate so as to control opening of a sealing cover piece and enable an opening of each test cavity to be in an open state;
step three, the rotating shaft rod 43 is rotated to drive the pushing piece to push Cry1Ab feed in each reagent tube to be thrown from the opening of each test cavity, so that white moth larvae in each test cavity can eat, the rotating shaft rod 43 stops rotating, the locking seat 6 is rotated to disconnect one end of the rotating shaft rod 43 from the rotating piece, and the locking seat 6 drives the swinging frame 52 to rotate to control the closing of the sealing cover piece, so that the opening of each test cavity is in a closed state;
Setting an observation period, raising the white moth larvae at the temperature of 27+/-2 ℃ and the relative humidity of 60% -70% and the photoperiod of 14 L:10D, observing the survival rate of the fed white moth larvae, and judging the sensitivity of the fall webworm to Cry1Ab toxins.
The artificial feed comprises, by weight, 5-12% of embryo bud, 2-5% of sucrose, 3-7% of protein, 0.3-1% of Welch salt, 0.4-1.2% of sorbic acid, 0.1-0.5% of methyl parahydroxybenzoate, 0.02-0.19% of ascorbic acid, 0.03-0.15% of vitamin B, 1.1-1.8% of agar, 0.05-0.16% of cholesterol and the balance of water, wherein Cry1Ab is MVPII aqua with 20% of Cry1 Ab-containing Bt ICP.
The foregoing examples illustrate only a few embodiments of the invention and are described in detail herein without thereby limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention.
Claims (8)
1. A biological assay device for detecting sensitivity of fall webworm to Cry1Ab toxin is characterized by comprising a test box (1), wherein four test cavities with upward openings are equally divided in the test box (1) by a cross baffle (2);
The test device comprises a sealing cover assembly (5) arranged on a test box (1) and a reagent tube group (3) clamped on the sealing cover assembly (5), wherein the number of the reagent tubes of the reagent tube group (3) is consistent with that of the test cavities;
the driving assembly (4) is arranged right above the reagent tube group (3), the driving assembly (4) comprises a rotating shaft rod (43) and a pushing piece arranged on the rotating shaft rod (43), and the pushing piece is in abutting sliding fit with the tube wall of each reagent tube of the reagent tube group (3) under the pushing of the rotating shaft rod (43);
The sealing cover assembly (5) comprises a swing frame (52) and a sealing cover piece, the outer side of the swing frame (52) is connected with a locking seat (6), a rotating piece is arranged in the locking seat (6), the locking seat (6) is rotated to enable one end of a rotating shaft rod (43) to be connected with the rotating piece, and the locking seat (6) drives the swing frame (52) to rotate to control opening of the sealing cover piece.
2. The bioassay device for detecting sensitivity of fall webworm to Cry1Ab toxin according to claim 1, wherein the structure of the single reagent tube in the reagent tube group (3) comprises an outer tube body (31), a conical tube (32) arranged at the lower end of the outer tube body (31) and a jet nozzle (33) arranged at the lower end of the conical tube (32).
3. The biological assay device for detecting sensitivity of fall webworm to Cry1Ab toxin according to claim 1 is characterized in that the pushing piece comprises a transverse frame (41), a nut seat (44) arranged on the transverse frame (41) and a threaded sleeve (46) sleeved on a rotating shaft rod (43) and in threaded fit with the nut seat (44), pushing rods (42) corresponding to the number of reagent tubes of the reagent tube group (3) are arranged on the lower end face of the transverse frame (41), and the upper end of the rotating shaft rod (43) is driven to rotate by a connecting driving motor (45).
4. The biological assay device for detecting sensitivity of fall webworm to Cry1Ab toxin according to claim 3 is characterized in that the locking seat (6) comprises an axle seat (61), one end of the rotating shaft rod (43) extending into the axle seat (61) is provided with an external thread part (48), the inner side wall of the axle seat (61) is provided with an internal thread part (64) matched with the external thread part (48), the rotating member comprises a ball seat (62) arranged in the axle seat (61) and a ball block (63) movably arranged in the ball seat (62), the lower end of the rotating shaft rod (43) is provided with a plugging rod (49) plugged with the ball block (63), the outside of the rotating shaft rod (43) is provided with a shaft sleeve (47), and the meshing transmission distance between the external thread part (48) of the rotating shaft rod (43) and the internal thread part (64) is equal to the plugging distance between the plugging rod (49) and the ball block (63).
5. The biological assay device for detecting sensitivity of fall webworm to Cry1Ab toxin according to claim 1 is characterized in that the sealing cover piece comprises a top cover (51), through holes corresponding to the positions of the test cavities are formed in the top cover (51), four partition frames (54) are uniformly distributed on the top cover (51), a partition cover (55) is rotatably matched inside the partition frames (54), a swing rod (53) connected with the partition cover (55) is arranged on the outer side of the swing frame (52) corresponding to the outer side of the partition frames (54), and a partition net (56) corresponding to the through holes is arranged on the partition cover (55).
6. The biological assay device for detecting the sensitivity of fall webworm to Cry1Ab toxin according to claim 5, characterized in that the spacer frame (54) comprises a frame plate (541), a rotating shaft (542) is rotatably fitted inside the frame plate (541) by providing a torsion spring (543), and one end of the spacer cover (55) is connected to the rotating shaft (542) and wound around the rotating shaft (542).
7. The biological assay device for detecting sensitivity of fall webworm to Cry1Ab toxin according to claim 1 is characterized in that a fecal accumulating plate (7) is embedded at the lower end of the test box (1), and a feeding plate (8) is arranged in each test cavity of the test box (1).
8. A method of detecting sensitivity of fall webworm to Cry1Ab toxins using the bioassay device of any one of claims 1-7, comprising the steps of,
Step one, putting the white moth larvae grouped according to the same quantity into each test cavity of a test box correspondingly, and keeping the white moth larvae fed under the same feeding environment;
Step two, preparing artificial feed containing Cry1Ab with different concentrations, taking the same amount of the artificial feed and respectively placing the same amount of artificial feed into each reagent tube of the reagent tube group (3), enabling one end of a rotating shaft rod (43) to extend into a locking seat (6) so that a pushing piece correspondingly extends into the mouth of each reagent tube, rotating the locking seat (6) to enable one end of the rotating shaft rod (43) to be connected with the rotating piece, and enabling the locking seat (6) to drive a swinging frame (52) to rotate so as to control opening of a sealing cover piece, so that an opening of each test cavity is in an open state;
Step three, driving a pushing part to push Cry1Ab feed in each reagent tube to be thrown from the opening of each test cavity after rotating a rotating shaft rod (43), feeding white moth larvae in each test cavity, stopping rotating the rotating shaft rod (43), rotating a locking seat (6), and driving a swinging frame (52) to rotate while one end of the rotating shaft rod (43) is disconnected from the rotating part, so as to control the closing of a sealing cover part, and enabling the opening of each test cavity to be in a closed state;
And step four, setting an observation period, observing the survival rate of the white moth larvae after feeding, and judging the sensitivity of the fall webworms to Cry1Ab toxins.
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| CN106596525A (en) * | 2016-12-30 | 2017-04-26 | 深圳市世纪天源环保技术有限公司 | Water biological toxicity detection method and device |
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| CN106596525A (en) * | 2016-12-30 | 2017-04-26 | 深圳市世纪天源环保技术有限公司 | Water biological toxicity detection method and device |
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