CN109566543A - A method of improving Candle-sticks stinkbug egg laying amount - Google Patents
A method of improving Candle-sticks stinkbug egg laying amount Download PDFInfo
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- CN109566543A CN109566543A CN201811619299.3A CN201811619299A CN109566543A CN 109566543 A CN109566543 A CN 109566543A CN 201811619299 A CN201811619299 A CN 201811619299A CN 109566543 A CN109566543 A CN 109566543A
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- 241000578422 Graphosoma lineatum Species 0.000 title claims abstract description 131
- 230000017448 oviposition Effects 0.000 title claims abstract description 37
- 238000000034 method Methods 0.000 title claims abstract description 34
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 115
- 102000002322 Egg Proteins Human genes 0.000 claims abstract description 76
- 108010000912 Egg Proteins Proteins 0.000 claims abstract description 76
- 210000004681 ovum Anatomy 0.000 claims abstract description 76
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 claims abstract description 44
- 230000009514 concussion Effects 0.000 claims abstract description 42
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 37
- 238000012545 processing Methods 0.000 claims abstract description 29
- 230000012447 hatching Effects 0.000 claims abstract description 21
- 230000001954 sterilising effect Effects 0.000 claims abstract description 19
- 239000008367 deionised water Substances 0.000 claims abstract description 17
- 229910021641 deionized water Inorganic materials 0.000 claims abstract description 17
- 238000009395 breeding Methods 0.000 claims description 10
- 239000007921 spray Substances 0.000 claims description 10
- 239000003153 chemical reaction reagent Substances 0.000 claims description 9
- 239000012153 distilled water Substances 0.000 claims description 6
- 241000819999 Nymphes Species 0.000 claims description 5
- 230000001488 breeding effect Effects 0.000 claims description 5
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 claims description 4
- 238000005119 centrifugation Methods 0.000 claims description 4
- 241000409991 Mythimna separata Species 0.000 claims description 2
- 230000001476 alcoholic effect Effects 0.000 claims 1
- 238000002474 experimental method Methods 0.000 abstract description 6
- 235000019441 ethanol Nutrition 0.000 description 33
- 235000013601 eggs Nutrition 0.000 description 11
- 238000004140 cleaning Methods 0.000 description 7
- 230000006378 damage Effects 0.000 description 7
- 230000000694 effects Effects 0.000 description 7
- 238000004659 sterilization and disinfection Methods 0.000 description 6
- 241000238631 Hexapoda Species 0.000 description 5
- 230000000052 comparative effect Effects 0.000 description 4
- 239000007788 liquid Substances 0.000 description 3
- 230000010355 oscillation Effects 0.000 description 3
- 238000012360 testing method Methods 0.000 description 3
- 241000607479 Yersinia pestis Species 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000575 pesticide Substances 0.000 description 2
- 241001124076 Aphididae Species 0.000 description 1
- 241000894006 Bacteria Species 0.000 description 1
- 241001367013 Ctenoplusia agnata Species 0.000 description 1
- 241000255967 Helicoverpa zea Species 0.000 description 1
- 241000254043 Melolonthinae Species 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 210000001136 chorion Anatomy 0.000 description 1
- 238000004821 distillation Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000003292 glue Substances 0.000 description 1
- 210000003128 head Anatomy 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 239000012535 impurity Substances 0.000 description 1
- 230000000266 injurious effect Effects 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 210000000554 iris Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000002035 prolonged effect Effects 0.000 description 1
- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 1
- 230000000384 rearing effect Effects 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
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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
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The present invention relates to a kind of method for improving Candle-sticks stinkbug egg laying amount, include the following steps: that 1) the new Candle-sticks stinkbug ovum that produces is put into centrifuge tube；2) after ethanol solution being added into the centrifuge tube in step 1), centrifuge tube is put into oscillator and carries out the processing of concussion three times；3) after formalin being added to the centrifuge tube in step 2), centrifuge tube is put into the concussion processing carried out in oscillator；4) into the centrifuge tube in step 3) after ethanol solution, centrifuge tube is put into oscillator and carries out concussion processing；5) by Candle-sticks stinkbug ovum obtained in step 4) with sterile deionized water be rinsed it is clean after, with blotting surface moisture in sterilizing filter paper.The present invention under the normal husbandry conditions of laboratory, effectively increases egg laying amount after hatching forms adult by the Candle-sticks stinkbug ovum that above-mentioned experimental procedure is handled, so that the expansion for facilitating Candle-sticks stinkbug population is numerous, saves economy, time and the cost of labor of the raising of Candle-sticks stinkbug.
Description
Technical field
The present invention relates to insect artificial feeding field more particularly to a kind of methods for improving Candle-sticks stinkbug egg laying amount.
Background technique
The use of long-term chemical pesticide seriously threatens China's food and ecological safety, has also greatly affected state
People's health, the prevention and treatment problem of crop pests is particularly important here.It can for the biological control of core currently, being used with natural enemy insect
The effectively quantity of control pest, reduces the use of pesticide.And the numerous work of interior expansion for carrying out natural enemy insect is that it is applied to nocuousness
The primary work of biological control.Candle-sticks stinkbug belongs to Semiptera Pentatomiddae, is a kind of wide natural enemy insect of predation range, can prey on it is chrysomelid,
The larva or adult of more than 40 agriculture and forestry injurious insect such as bollworm, three-spotted plusia, slug moth, chafer, aphid.Since Candle-sticks stinkbug is by indoor
Mostly after generation raising, it often will appear egg laying amount decline, ovum reduces etc. again and to be unfavorable for its growth and development and expand numerous phenomenon.Oviposition
Amount often will affect Candle-sticks stinkbug population expansion is numerous and economy, time and the cost of labor of raising, how to improve the egg laying amount of Candle-sticks stinkbug at
For the matter of utmost importance of Candle-sticks stinkbug breeding process.
Summary of the invention
Compared with prior art, it the present invention provides a kind of method for improving Candle-sticks stinkbug egg laying amount, is asked with solving the prior art
At least one of topic technical problem.
The technical scheme to solve the above technical problems is that a kind of method for improving Candle-sticks stinkbug egg laying amount, including such as
Lower step: 1) new produce Candle-sticks stinkbug ovum is put into centrifuge tube；
2) after ethanol solution being added into the centrifuge tube in step 1), centrifuge tube is put into oscillator and is shaken three times
Processing；2.5~3.5min of centrifuge tube is stood between the adjacent concussion processing described twice；Then the ethanol solution in centrifuge tube is inhaled
Out；
3) after formalin being added to the centrifuge tube in step 2), centrifuge tube is put at the concussion carried out in oscillator
Reason；After standing 4~5min, the formalin in the centrifuge tube is sucked out；
4) into the centrifuge tube in step 3) after ethanol solution, centrifuge tube is put into oscillator and carries out concussion processing；It is quiet
After setting 3~4min, the ethanol solution in the centrifuge tube is sucked out；
5) after being rinsed Candle-sticks stinkbug ovum obtained in step 4) completely with sterile deionized water, Candle-sticks is blotted with sterilizing filter paper
Stinkbug ovum surface moisture.
Based on the above technical solution, the present invention can also be improved as follows:
Preferably, the volume that Candle-sticks stinkbug ovum is newly produced described in step 1) accounts for the 1/3~2/5 of centrifugation pipe volume；In step 2), to
It is added in centrifuge tube in step 1) after not crossing the ethanol solution that the volumetric concentration of Candle-sticks stinkbug ovum is 75%, centrifuge tube is put into concussion
The processing of concussion three times is carried out in device, is every time that 500-1000rpm shakes 13~17s with revolving speed, between adjacent concussion twice processing
Stand 2.5~3.5min of centrifuge tube；Then the ethanol reagent in centrifuge tube is sucked out.
Preferably, in step 3), being added to the centrifuge tube in step 2) and not crossing the volumetric concentration of Candle-sticks stinkbug ovum is that 4% formaldehyde is molten
After liquid, centrifuge tube is put into 13~17s of progress in oscillator, the concussion that revolving speed is 500-1000rpm is handled, and stands 4~5min
Afterwards, the formalin in the centrifuge tube is sucked out.
Preferably, in step 4), being added into the centrifuge tube in step 3) and not crossing the volumetric concentration of Candle-sticks stinkbug ovum is 75%
After ethanol solution, the centrifuge tube is put into 13~17s of progress in oscillator, the concussion that revolving speed is 500-1000rpm is handled,
After standing 3~4min, the ethanol solution in the centrifuge tube is sucked out.
Preferably, in step 5), after Candle-sticks stinkbug ovum obtained in step 4) is rinsed completely with sterile deionized water, then
Candle-sticks stinkbug oophyte is placed in sterilizing filter paper and blots surface moisture.
Preferably, the method for improving Candle-sticks stinkbug egg laying amount further includes step 6), and Candle-sticks stinkbug ovum obtained in step 5) is equal
Even is placed in sterilizes culture dish, and the temperature that culture environment is arranged is 25~27, DEG C humidity is 60~70%, the photoperiod is
15L:9D~17L:7D；It is placed with the sterilizing filter paper for having sprayed sterile purified water in the culture dish, sprays one into culture dish daily
Secondary sterile purified water, until Candle-sticks stinkbug egg hatching.
Preferably, the Candle-sticks stinkbug nymph after hatching obtained in step 6) is subjected to laboratory conventinal breeding, the laboratory
Conventinal breeding method includes feeding distilled water and tussah chrysalis and/or mythimna separata.
Preferably, the photoperiod of the environment of culture Candle-sticks stinkbug ovum is 16L:8D in step 6).
Preferably, the oscillator of step 2), step 3) and step 4) is turbula shaker；Step 2), step 3) and step
It is rapid 4) in by centrifuge tube reagent be sucked out tool be pipette.
Preferably, 3~5 grams of new Candle-sticks stinkbug ovum that produce are put into 50ml centrifuge tube by step 1)；Step 2), step 3) and step
4) it is respectively 15s that centrifuge tube, which is put into the time that oscillator shakes, in；Oscillator in step 2), step 3) and step 4)
Revolving speed be respectively 700~800rpm；Ethanol solution in step 2) and step 4) can be replaced volumetric concentration be 70% it is different
Propanol solution.
In conclusion the beneficial effects of the present invention are: the Candle-sticks stinkbug ovum handled by above-mentioned experimental procedure, hatching formed at
After worm, under the normal husbandry conditions of laboratory, egg laying amount is effectively increased, so that the expansion for facilitating Candle-sticks stinkbug population is numerous, saves Candle-sticks
Economy, time and the cost of labor of stinkbug raising.
Specific embodiment
Principles and features of the present invention are described with reference to embodiments, the given examples are served only to explain the present invention,
It is not intended to limit the scope of the present invention.
The present invention relates to a kind of method for improving Candle-sticks stinkbug egg laying amount, include the following steps: 1) to be put into the new Candle-sticks stinkbug ovum that produces
In centrifuge tube；2) after ethanol solution being added into the centrifuge tube in step 1), centrifuge tube is put into oscillator and is shaken three times
Swing processing；2.5~3.5min of centrifuge tube is stood between the adjacent concussion processing described twice；Then the ethyl alcohol in centrifuge tube is tried
Agent is sucked out；3) after formalin being added to the centrifuge tube in step 2), centrifuge tube is put into the concussion processing carried out in oscillator；
4~5min is stood, then the formalin in the centrifuge tube is sucked out；4) ethanol solution into the centrifuge tube in step 3)
Afterwards, centrifuge tube is put into oscillator and carries out concussion processing；It is then that the ethyl alcohol in the centrifuge tube is molten after standing 3~4min
Liquid is sucked out；5) after being rinsed Candle-sticks stinkbug ovum obtained in step 4) completely with sterile deionized water, Candle-sticks is blotted with sterilizing filter paper
Stinkbug ovum surface moisture.The Candle-sticks stinkbug ovum that the present invention is handled by above-mentioned experimental procedure, after hatching forms adult, in laboratory routine
Under rearing conditions, effectively increase egg laying amount, so that the expansion for helping to improve Candle-sticks stinkbug population is numerous, save the raising of Candle-sticks stinkbug economy,
Time and cost of labor.The method of the application protection can continue to apply in the oophyte that a generation generates, and Candle-sticks stinkbug is continuously improved
Egg laying amount.
In some embodiments, a method of improving Candle-sticks stinkbug egg laying amount, include the following steps: 1) to put the new Candle-sticks stinkbug ovum that produces
Enter into centrifuge tube；The new volume for producing Candle-sticks stinkbug ovum accounts for the 1/3~2/5 of centrifugation pipe volume；2) add into the centrifuge tube in step 1)
After the ethanol solution that the volumetric concentration for entering not cross Candle-sticks stinkbug ovum is 75%, centrifuge tube is put into oscillator and is carried out at concussion three times
Reason is every time that 500-1000rpm shakes 13~17s with revolving speed, stood between adjacent concussion twice processing centrifuge tube 2.5~
3.5min；Then the ethanol reagent in centrifuge tube is sucked out；3) volume for not crossing Candle-sticks stinkbug ovum is added to the centrifuge tube in step 2)
For concentration centrifuge tube to be put into oscillator and carries out 13~17s after 4% formalin, revolving speed is at the concussion of 500-1000rpm
After standing 4~5min, then the formalin in the centrifuge tube is sucked out for reason；4) it is added into the centrifuge tube in step 3)
After the ethanol solution that the volumetric concentration for not crossing Candle-sticks stinkbug ovum is 75%, the centrifuge tube is put into 13~17s of progress in oscillator, is turned
The concussion that speed is 500-1000rpm is handled, and after standing 3~4min, then the ethanol solution in the centrifuge tube is sucked out；5)
By Candle-sticks stinkbug ovum obtained in step 4) with sterile deionized water be rinsed it is clean after, then Candle-sticks stinkbug oophyte is placed on sterilizing filter paper
On blot surface moisture；6) Candle-sticks stinkbug ovum obtained in step 5) is uniformly placed in sterilizes culture dish, culture environment is set
Temperature be 25~27, DEG C humidity is 60~70%, the photoperiod is 15L:9D~17L:7D；It is placed with and sprays in the culture dish
The sterilizing filter paper of sterile purified water sprays a sterile purified water into culture dish daily, until Candle-sticks stinkbug egg hatching.It is new to produce Candle-sticks stinkbug ovum
Oophyte generally within 12 to 24 hours postpartum.
In the method for above-mentioned raising Candle-sticks stinkbug egg laying amount, the volume that Candle-sticks stinkbug ovum is newly produced in step 1) accounts for the 1/ of centrifugation pipe volume
3~2/5, it is ensured that the maximum quantity of the Candle-sticks stinkbug ovum of single treatment improves treatment effeciency；
In step 2), the ethanol solution that volumetric concentration is 75% is added into the centrifuge tube in step 1), for Candle-sticks stinkbug
Ovum surface carries out disinfection, and since excessive ethanol solution may cause the damage to Candle-sticks stinkbug ovum, the ethanol solution being added does not have
Candle-sticks stinkbug ovum is crossed, the application does not do excessive limitation to the ethanol solution amount that centrifuge tube is added.Centrifuge tube is put into oscillator
Oophyte surface can be effectively facilitated and come into full contact with reagent by carrying out concussion processing, selection 500~1000rpm revolving speed in method,
Oophyte processing is carried out within the scope of this impact strength can achieve good cleaning effect, while oophyte will not be made to rupture；Every time
Concussion duration is set as 13~17s, while repeating repeatedly to shake, and can achieve preferably cleaning effect, effectively avoids out
Now it is located at the ovum above centrifuge tube to be cleaned, and the ovum of bottom does not reach cleaning effect；Per concussion processing twice
Between be arranged a period of time standing, effectively avoid for a long time shake mechanical damage is brought to oophyte.
In step 3), the formalin that volumetric concentration is 4% is added into the centrifuge tube in step 2), for Candle-sticks stinkbug ovum
Surface carries out disinfection, and since excessive formalin may cause the damage to Candle-sticks stinkbug ovum, the formalin being added did not had
Candle-sticks stinkbug ovum, the application do not do excessive limitation to the formalin amount that centrifuge tube is added.By centrifuge tube be put into oscillator into
Row concussion processing can effectively facilitate oophyte surface and come into full contact with reagent, 500~1000rpm revolving speed be selected in method, at this
Oophyte processing is carried out within the scope of one impact strength can achieve good cleaning effect, while oophyte will not be made to rupture；When concussion
Length is set as 13~17s, can preferably clean oophyte；The standing of a period of time is set after earthquake, effectively restores concussion
Mechanical damage is brought to oophyte.It should be noted that selecting formalin in step 3), effectively avoid micro- on step 2) oophyte
The resistance for ethyl alcohol that biology is formed in the short time, effectively increases the effect of disinfection and sterilization；Since ethyl alcohol is easy to make biological group
It knits and becomes fragile, prolonged impregnate may bring injury to chorion, influence to hatch, and select formalin that can reduce here this
Risk.
In step 4), the ethanol solution that volumetric concentration is 75% is added into the centrifuge tube in step 3), for Candle-sticks stinkbug
Ovum surface carries out disinfection, and since excessive ethanol solution may cause the damage to Candle-sticks stinkbug ovum, the ethanol solution being added does not have
Candle-sticks stinkbug ovum is crossed, the application does not do excessive limitation to the ethanol solution amount that centrifuge tube is added.Centrifuge tube is put into oscillator
Oophyte surface can be effectively facilitated and come into full contact with reagent by carrying out concussion processing, selection 500~1000rpm revolving speed in method,
Oophyte processing is carried out within the scope of this impact strength can achieve good cleaning effect, while oophyte will not be made to rupture；Concussion
Duration is set as 13~17s, can preferably clean oophyte；The standing of a period of time is set after earthquake, effectively restores shake
It swings and mechanical damage is brought to oophyte.
In step 5), by Candle-sticks stinkbug ovum obtained in step 4) with sterile deionized water be rinsed it is clean after, then by Candle-sticks stinkbug
Oophyte, which is placed in sterilizing filter paper, blots surface moisture；Oophyte is rinsed with sterile deionized water again, can be removed residual
The ethanol solution stayed；
In step 6), Candle-sticks stinkbug ovum obtained in step 5) is uniformly placed in sterilizes culture dish, culture environment is set
Temperature be 25~27, DEG C humidity is 60~70%, the photoperiod is 15L:9D~17L:7D；It is placed with and sprays in the culture dish
The sterilizing filter paper of sterile purified water, sprays a sterile purified water into culture dish daily, until Candle-sticks stinkbug egg hatching.
Step 2)~5) in carried out multiple washing, concussion, facilitate by cleaning generate microbial impurities clears up in time,
To reach better experiment effect.The Candle-sticks stinkbug nymph after hatching obtained in step 6) is subjected to laboratory conventinal breeding later.
In some embodiments, the laboratory conventinal breeding method includes feeding distilled water and tussah chrysalis and/or glues
Worm.In step 6) by Candle-sticks stinkbug ovum it is smooth one down be placed on 12cm sterilizing glass culture dish in.It should be noted that Candle-sticks stinkbug
The oophyte shape of ovum is oval structures, and one end end face border of Candle-sticks stinkbug ovum is with outside small burr, a Duan Duan on the other side
Face is smooth, does not in addition grow other structures, and there is the opening of the broken ovum of hatching in long jagged one end end face, and Candle-sticks stinkbug nymph opens
Egg apparatus can puncture spinule, and that irises out, therefore smooth face-down, facilitates the hatching of Candle-sticks stinkbug ovum, the hatching of ovum can be improved
Rate.The oscillator of step 2), step 3) and step 4) is turbula shaker.Turbula shaker is easily operated and regulates and controls, certainly,
The application is not limited to vortex oscillator；The tool that the reagent in centrifuge tube is sucked out in step 2), step 3) and step 4) is to move
Liquid pipe, pipette is easily operated and regulates and controls, and certainly, the application is not limited to pipette.
In one or more embodiments, centrifuge tube oscillator is put into step 2), step 3) and step 4) to shake
The time swung be 15s, after test of many times, duration of oscillation it is longer or it is shorter be not well positioned to meet mixing disinfection needs,
Duration of oscillation is that the 15s time is preferable；Preferably, in step 2), step 3) and step 4) oscillator revolving speed be 700~
800rpm, after test of many times, oscillation intensity is smaller or crosses the needs that cannot mostly meet mixing cleaning well, vibrates revolving speed
It is preferable for 700~800rpm time.
In some embodiments, since specificity of reagent is close, the ethanol solution in step 2) and step 4) can be replaced body
The aqueous isopropanol that product concentration is 70%.
In at least one embodiment, the photoperiod that the environment of Candle-sticks stinkbug ovum is cultivated in step 6 is 16L:8D, the illumination of oophyte
Period is extremely important to oophyte development, obtains through test of many times, and the photoperiod is Candle-sticks stinkbug hatching within the scope of 15L:9D~17L:7D
Rate is higher, and the photoperiod be 16L:8D when, hatching rate is best.
The Candle-sticks stinkbug ovum handled by above-mentioned experimental procedure, after hatching forms adult, under the normal husbandry conditions of laboratory,
Egg laying amount is effectively increased, so that the expansion for improving Candle-sticks stinkbug population is numerous, saves economy, time and the cost of labor of the raising of Candle-sticks stinkbug.
It is illustrated below by way of specific embodiment:
Embodiment 1
A method of Candle-sticks stinkbug egg laying amount is improved, includes the following steps: that 3~5 grams of new Candle-sticks stinkbug ovum that produce 1) are put into 50ml
In centrifuge tube；2) it is added into the centrifuge tube in step 1) after not crossing the ethanol solution that the volumetric concentration of Candle-sticks stinkbug ovum is 75%, it will
Centrifuge tube, which is put into oscillator, carries out the processing of concussion three times, is every time that 700~800rpm shakes 15s with revolving speed, adjacent to shake twice
It swings and stands centrifuge tube 3min between processing；Then the ethanol solution in centrifuge tube is sucked out；3) add to the centrifuge tube in step 2)
Enter do not cross Candle-sticks stinkbug ovum volumetric concentration be 4% formalin after, centrifuge tube is put into oscillator and carries out 15s, revolving speed 700
The concussion of~800rpm is handled, and after standing 4min, then the formalin in the centrifuge tube is sucked out；4) into step 3)
Be added in centrifuge tube do not cross Candle-sticks stinkbug ovum volumetric concentration be 75% ethanol solution after, by the centrifuge tube be put into oscillator into
Row 15s, the concussion that revolving speed is 700~800rpm is handled, and after standing 4min, is then inhaled the ethanol solution in the centrifuge tube
Out；5) after being rinsed Candle-sticks stinkbug ovum obtained in step 4) completely with sterile deionized water, then Candle-sticks stinkbug oophyte is placed on and is gone out
Surface moisture is blotted on bacterium filter paper；6) Candle-sticks stinkbug ovum obtained in step 5) is uniformly placed in 12cm sterilizes culture dish, if
The temperature for setting culture environment is 25~27, DEG C humidity is 60~70%, photoperiod 16L:6D；Spray is placed in the culture dish
The sterilizing filter paper of sterile purified water has been applied, has sprayed a sterile purified water into culture dish daily, until Candle-sticks stinkbug egg hatching.Here it sets
It sets and is set as EG group by the processed oophyte of the above method.
Comparative example 1
Include the following steps: that 1) 3~5 grams of new Candle-sticks stinkbug ovum that produce are put into 50ml centrifuge tube；2) in step 1) from
It is added in heart pipe after not crossing Candle-sticks stinkbug ovum sterile deionized water, centrifuge tube is put into oscillator and carries out the processing of concussion three times, every time
It is that 700~800rpm shakes 15s with revolving speed, stands centrifuge tube 3min between the adjacent processing of concussion twice；It then will be in centrifuge tube
Sterile deionized water be sucked out；3) it after the sterile deionized water for not crossing Candle-sticks stinkbug ovum is added to the centrifuge tube in step 2), will be centrifuged
Pipe, which is put into oscillator, carries out 15s, and the concussion that revolving speed is 700~800rpm is handled, after standing 4min, then by the centrifuge tube
In sterile deionized water be sucked out；It 4), will after the sterile deionized water for not crossing Candle-sticks stinkbug ovum is added into the centrifuge tube in step 3)
The centrifuge tube, which is put into oscillator, carries out 15s, and the concussion that revolving speed is 700~800rpm is handled, and after standing 4min, then will
Sterile deionized water in the centrifuge tube is sucked out；5) Candle-sticks stinkbug ovum obtained in step 4) is rushed with sterile deionized water
After wash clean, then Candle-sticks stinkbug oophyte is placed in sterilizing filter paper and blots surface moisture；6) Candle-sticks stinkbug ovum obtained in step 5) is equal
Even is placed in 12cm sterilizes culture dish, and the temperature that culture environment is arranged is 25~27, DEG C humidity is 60~70%, the photoperiod
For 16L:6D；It is placed with the sterilizing filter paper for having sprayed sterile purified water in the culture dish, sprays primary sterilizing into culture dish daily
Distilled water, until Candle-sticks stinkbug egg hatching.Here setting is set as PG group by the processed oophyte of the above method.
Comparative example 2
Untreated new production Candle-sticks stinkbug 3~5g of ovum is taken, is uniformly placed in 12cm sterilizes culture dish, culture environment is set
Temperature be 25~27, DEG C humidity is 60~70%, photoperiod 16L:6D；It is placed in the culture dish and has sprayed aseptic distillation
The sterilizing filter paper of water sprays a sterile purified water into culture dish daily, until Candle-sticks stinkbug egg hatching.Here above-mentioned side is passed through in setting
The processed oophyte of method is set as CG group.
A sterile purified water is sprayed to EG group, PG group and CG group culture dish daily, until egg hatching, if the Candle-sticks stinkbug after hatching
Worm and adult carry out conventinal breeding in laboratory, and conventinal breeding method includes providing distilled water and tussah chrysalis, distilled water to Candle-sticks stinkbug
Add primary, replacement in tussah chrysalis 3~5 days daily once；The raising of nymph single head, the processes such as male pairing raising of adult one female one, due to
Conventinal breeding method is well-known technique, is not discussed here.The experiment of embodiment 1, comparative example 1 and comparative example 2 respectively into
5 repetitions of having gone are tested, and substantially avoid experimental implementation contingency, and it is right that each group respectively matches Candle-sticks stinkbug more than 400, randomly select 100
To record its lay eggs situation such as table 1.
1 Candle-sticks stinkbug ovum of table Candle-sticks stinkbug Adult worms producting eggs situation statistical form after different disposal
As seen from Table 1: the egg laying amount of PG significantly rises compared with CG, and the egg laying amount of EG significantly rises compared with CG, PG
(F2,297=14.93, P < 0.0001), the same trend (F2,297=14.56, P < 0.0001) that oviposition number is also presented are single
Secondary egg laying amount is without significant difference.EG egg laying amount improves 56.15% relative to CG, and egg hatching rate has decline by a small margin but without aobvious
It writes difference (F2,297=2.878, P=0.0578).It proves that this method can improve Candle-sticks stinkbug egg laying amount well, and will not influence
The single egg laying amount of Candle-sticks stinkbug.
In the description of this specification, reference term " one embodiment ", " some embodiments ", " example ", " specifically show
The description of example " or " some examples " etc. means specific features, structure, material or spy described in conjunction with this embodiment or example
Point is included at least one embodiment or example of the invention.In the present specification, schematic expression of the above terms are not
It must be directed to identical embodiment or example.Moreover, particular features, structures, materials, or characteristics described can be in office
It can be combined in any suitable manner in one or more embodiment or examples.In addition, without conflicting with each other, the skill of this field
Art personnel can tie the feature of different embodiments or examples described in this specification and different embodiments or examples
It closes and combines.
The foregoing is merely presently preferred embodiments of the present invention, is not intended to limit the invention, it is all in spirit of the invention and
Within principle, any modification, equivalent replacement, improvement and so on be should all be included in the protection scope of the present invention.
Claims (10)
1. a kind of method for improving Candle-sticks stinkbug egg laying amount, which comprises the steps of:
1) the new Candle-sticks stinkbug ovum that produces is put into centrifuge tube；
2) after ethanol solution being added into the centrifuge tube in step 1), the centrifuge tube is put into oscillator and is carried out three times
Concussion processing；2.5~3.5min of centrifuge tube is stood between the adjacent concussion processing described twice；Then by the second in the centrifuge tube
Alcoholic solution is sucked out；
3) after formalin being added to the centrifuge tube in step 2), the centrifuge tube is put into the concussion carried out in oscillator
Processing；After standing 4~5min, the formalin in the centrifuge tube is sucked out；
4) centrifuge tube is put into oscillator and is carried out at concussion after ethanol solution into the centrifuge tube in step 3)
Reason；After standing 3~4min, the ethanol solution in the centrifuge tube is sucked out；
5) after being rinsed Candle-sticks stinkbug ovum obtained in step 4) completely with sterile deionized water, Candle-sticks stinkbug ovum is blotted with sterilizing filter paper
Surface moisture.
2. a kind of method for improving Candle-sticks stinkbug egg laying amount according to claim 1, which is characterized in that newly produce Candle-sticks described in step 1)
The volume of stinkbug ovum accounts for the 1/3~2/5 of centrifugation pipe volume；In step 2), it is added into the centrifuge tube in step 1) and does not cross Candle-sticks
After the ethanol solution that the volumetric concentration of stinkbug ovum is 75%, the centrifuge tube is put into oscillator and carries out the processing of concussion three times, often
Secondary with revolving speed is that 500-1000rpm shakes 13~17s, stood between adjacent concussion twice processing the centrifuge tube 2.5~
3.5min；Then the ethanol solution in centrifuge tube is sucked out.
3. a kind of method for improving Candle-sticks stinkbug egg laying amount according to claim 1, which is characterized in that in step 3), to step 2)
In the centrifuge tube be added do not cross Candle-sticks stinkbug ovum volumetric concentration be 4% formalin after, the centrifuge tube is put into oscillator
13~17s of middle progress, the concussion that revolving speed is 500-1000rpm is handled, after standing 4~5min, by the formaldehyde in the centrifuge tube
Solution is sucked out.
4. a kind of method for improving Candle-sticks stinkbug egg laying amount according to claim 1, which is characterized in that in step 4), to step 3)
In the centrifuge tube in be added do not cross Candle-sticks stinkbug ovum volumetric concentration be 75% ethanol solution after, the centrifuge tube is put into vibration
13~17s of progress in device is swung, the concussion that revolving speed is 500-1000rpm is handled, will be in the centrifuge tube after standing 3~4min
Ethanol solution is sucked out.
5. a kind of method for improving Candle-sticks stinkbug egg laying amount according to claim 1, which is characterized in that in step 5), by step 4)
Obtained in Candle-sticks stinkbug ovum with sterile deionized water be rinsed it is clean after, then Candle-sticks stinkbug oophyte is placed in sterilizing filter paper and blots table
Face moisture.
6. a kind of method for improving Candle-sticks stinkbug egg laying amount according to claim 1, which is characterized in that the raising Candle-sticks stinkbug egg laying amount
Method further include step 6), Candle-sticks stinkbug ovum obtained in step 5) is uniformly placed in sterilizes culture dish, setting culture ring
The temperature in border is 25~27 DEG C, humidity is 60~70%, the photoperiod is 15L:9D~17L:7D；It is placed with and sprays in the culture dish
The sterilizing filter paper of sterile purified water, sprays a sterile purified water into culture dish daily, until Candle-sticks stinkbug egg hatching.
7. a kind of method for improving Candle-sticks stinkbug egg laying amount according to claim 2, which is characterized in that will be incubated obtained in step 6)
Candle-sticks stinkbug nymph after change carries out laboratory conventinal breeding, and the laboratory conventinal breeding method includes feeding distilled water and tussah chrysalis
And/or mythimna separata.
8. a kind of method for improving Candle-sticks stinkbug egg laying amount according to claim 2, which is characterized in that cultivate Candle-sticks stinkbug ovum in step 6)
Environment photoperiod be 16L:8D.
9. a kind of method for improving Candle-sticks stinkbug egg laying amount according to claim 1, which is characterized in that step 2), step 3) and step
Rapid oscillator 4) is turbula shaker；The tool that the reagent in centrifuge tube is sucked out in step 2), step 3) and step 4)
It is pipette.
10. a kind of method for improving Candle-sticks stinkbug egg laying amount according to claim 1, which is characterized in that step 1) is new by 3~5 grams
Candle-sticks stinkbug ovum is produced to be put into 50ml centrifuge tube；Centrifuge tube oscillator is put into step 2), step 3) and step 4) to shake
Time be respectively 15s；The revolving speed of oscillator is respectively 700~800rpm in step 2), step 3) and step 4)；Step 2) and
Ethanol solution in step 4) can be replaced the aqueous isopropanol that volumetric concentration is 70%.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110476901A (en) * | 2019-09-17 | 2019-11-22 | 中国农业科学院植物保护研究所 | A method of extending Candle-sticks stinkbug ovum shelf life |
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