CN107568199A - The preparation method of Tianjin river amblyseius as predatory mite hero mite sperm slide sample - Google Patents
The preparation method of Tianjin river amblyseius as predatory mite hero mite sperm slide sample Download PDFInfo
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- CN107568199A CN107568199A CN201710808918.2A CN201710808918A CN107568199A CN 107568199 A CN107568199 A CN 107568199A CN 201710808918 A CN201710808918 A CN 201710808918A CN 107568199 A CN107568199 A CN 107568199A
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Abstract
The present invention relates to the preparation method of Tianjin river amblyseius as predatory mite hero mite sperm slide sample, it is characterized in that making the method for sperm slide sample after spermatophore is artificially obtained from Tianjin river amblyseius as predatory mite hero mite gnathosoma to mate, 5 parts are prepared comprising the acquisition of male and female mite, the sizing of male and female mite, the separation of male and female mite, the acquisition of spermatophore, the acquisition of sperm and slide sample in mating.The present invention is obtained with Tianjin river amblyseius as predatory mite hero mite spermatophore and is produced sperm slide sample using very simple cleverly method, the acquisition of spermatophore can provide raw material for the making and research of Tianjin river amblyseius as predatory mite sperm slide sample, and making for sperm slide sample can provide reference to understand the form of Tianjin river amblyseius as predatory mite sperm, disclosing Tianjin river amblyseius as predatory mite reproduction principle.
Description
Technical field
The present invention relates to the preparation method of Tianjin river amblyseius as predatory mite hero mite sperm slide sample, it is biologically-based to belong to Tianjin river amblyseius as predatory mite
Plinth research field.
Background technology
Tianjin river amblyseius as predatory mite is gonochorism animal, must can just be laid eggs by male and female mite mating after fertilization and breed offspring, offspring
No matter male and female are come from development of fertilized ova.
Available data mates for Tianjin river amblyseius as predatory mite male and female mite to be formed the understanding of embryonated egg and is：Male mite sperm is entering female mite
It is stored in before in vivo in spermatophore, spermatophore is filled in female mite copulatory opening by female mite（Bibliography：Liang Lairong, 1979, plant protection,
How mite class lives）, sperm comes out from spermatophore slow release again, is transferred to spermatheca and ovum combines, form embryonated egg.But
How to enter in female mite body for Tianjin river amblyseius as predatory mite hero mite sperm and male mite sperm is not reported with the presence or absence of dimorphism, if
Research and solve this series of problems, it is necessary first to obtain sperm, film-making observational study is carried out to sperm, and Sperm storage is in essence
In bag, it is therefore desirable to first obtain spermatophore and then carry out film-making observation to the sperm in spermatophore.
But due to very small only hundreds of microns of Tianjin river amblyseius as predatory mite individual, its spermatophore just goes out when there was only the mating of male and female mite
Existing, male and female mite being superimposed together closely in mating process, mating time is also very short only 1 hour or so, and spermatophore
Size only has more than ten micron and is hidden between the outside of belly of male and female mite body close contact, is intuitively hardly visible, existing money
The method for expecting also how not obtain spermatophore.Therefore want obtain spermatophore it is very difficult, make spermatophore slide sample it is more difficult, it is necessary to
Open up a kind of method for obtaining Tianjin river amblyseius as predatory mite spermatophore, making Tianjin river amblyseius as predatory mite sperm slide sample.
The content of the invention
It is an object of the invention to provide Tianjin river amblyseius as predatory mite hero mite sperm slide sample preparation method, using this method in body formula
Male mite spermatophore can be just obtained under microscope and produces sperm slide sample.
The present invention is to make sperm slide after spermatophore is artificially obtained from Tianjin river amblyseius as predatory mite hero mite gnathosoma to mate
The method of sample, include the acquisition of male and female mite in mating, the sizing of male and female mite, the separation of male and female mite, the acquisition of spermatophore, sperm
Obtain and slide sample prepares 5 parts, all operations are carried out under the microscope in addition to the male and female mite sizing in mating, specific behaviour
As：
（1）The acquisition of male and female mite in mating：Out of Tianjin river amblyseius as predatory mite raising population, the ovum for choosing Tianjin river amblyseius as predatory mite is individually raised extremely
Male and female mite pairing is carried out after into mite, 10 minutes or so male and female mites of pairing can mate, so as to male and female mite in being mated；
（2）The sizing of male and female mite in mating：In phytoseiid mite post-coitum 0-120min, by the male and female being in mating process
Body is positioned over 3-5min in -80 DEG C of ultra low temperature freezers or liquid nitrogen together with breeding apparatus, carries out male and female mite sizing；
（3）The separation of male and female mite after sizing：Male and female mite is taken out together with breeding apparatus after sizing, then with fine, soft fur pen from raising
The male and female mite of mating is chosen on slide in device, recycles the instrument such as No. 0 insect needle and fine, soft fur pen by the male and female mite of mating
Gently separate, prevent from destroying spermatophore；
（4）The acquisition of spermatophore：After male and female mite successfully separates, Hubei Province body of male mite is scaled off using scalpel, retains gnathosoma, in jaw
The spermatophore in spermatodactyl top is readily observed on body, then will be put into 70%-100% under the light picking of spermatophore with fine, soft fur nib
Preserve in alcohol or directly utilize；
（5）It is prepared by the acquisition of sperm and slide sample：The Klorvess Liquid of a drop 0.075mol/L is instilled in load with rubber head dropper
The middle position of slide, then Tianjin river amblyseius as predatory mite spermatophore is positioned in the Klorvess Liquid on slide with No. 0 fine, soft fur pen, often
Temperature is lower to be placed 30 minutes, then Klorvess Liquid is siphoned away or is transferred to spermatophore with fine, soft fur pen new with liquid-transfering gun or rubber head dropper
On slide, spermatophore is gently staved with No. 0 insect needle or thinner needle-shaped tool, and it is quick new toward the drop of instillation one on slide
The fixer of configuration（Methanol：Glacial acetic acid=3:1）, 30 minutes are fixed, at least 3 fixers are added dropwise in supplement in the process（Because
Fixer evaporation rate is fast）, after liquid to be fixed is dried, toward slide on instill Giemsa stain（1 part of stoste, PH7.2 phosphoric acid delays
9 parts of dilutions of fliud flushing）, dye 60 minutes under normal temperature, finally rinsed out dye liquor with clear water, through spontaneously drying or being dried up with hair-dryer
Afterwards, toward instilling an excellent Ba Laer of drop above the position that color is caught on slide, then covered, and with marking pen to dye
The position of colouring carries out circle note to facilitate observation, and i.e. observable Tianjin river amblyseius as predatory mite is male under an optical microscope after sample fixes
Mite sperm.
The present invention is obtained with Tianjin river amblyseius as predatory mite hero mite spermatophore and is produced sperm using very simple cleverly method
Slide sample, the acquisition of spermatophore can provide raw material, sperm slide mark for the making and research of Tianjin river amblyseius as predatory mite sperm slide sample
Making for this can provide reference to understand the form of Tianjin river amblyseius as predatory mite sperm, disclosing Tianjin river amblyseius as predatory mite reproduction principle.
Embodiment
The preparation method of embodiment Tianjin river amblyseius as predatory mite hero mite sperm slide sample
This method is the method for artificially obtaining from Tianjin river amblyseius as predatory mite hero mite gnathosoma to mate, choosing lower spermatophore, includes friendship
Acquisition, the sizing of male and female mite, the separation of male and female mite, the acquisition of spermatophore with middle male and female mite are with preserving 4 parts, and all operations are except friendship
Carried out under the microscope outside male and female mite sizing in matching somebody with somebody, concrete operations are：
（1）The acquisition of male and female mite in mating：Out of Tianjin river amblyseius as predatory mite raising population, the ovum for choosing Tianjin river amblyseius as predatory mite is individually raised extremely
Male and female mite pairing is carried out after into mite, 10 minutes or so male and female mites of pairing can mate, so as to male and female mite in being mated；
（2）The sizing of male and female mite in mating：In phytoseiid mite post-coitum 0-120min, by the male and female being in mating process
Body is positioned over 3-5min in -80 DEG C of ultra low temperature freezers or liquid nitrogen together with breeding apparatus, carries out male and female mite sizing；
（3）The separation of male and female mite after sizing：Male and female individual is taken out together with breeding apparatus after sizing, then with fine, soft fur pen from feeding
Support and choose the male and female mite of mating on slide in device, recycle the instrument such as No. 0 insect needle and fine, soft fur pen by the male and female of mating
Mite gently separates, and prevents from destroying spermatophore；
（4）The acquisition and preservation of spermatophore：After male and female mite successfully separates, Hubei Province body of male mite is scaled off using scalpel, retains jaw
Body, the spermatophore in spermatodactyl top is readily observed on gnathosoma, then will be put into fine, soft fur nib under the light picking of spermatophore
Preserve in 70%-100% alcohol or directly utilize；
（5）It is prepared by the acquisition of sperm and slide sample：The Klorvess Liquid of a drop 0.075mol/L is instilled in load with rubber head dropper
The middle position of slide, then Tianjin river amblyseius as predatory mite spermatophore is positioned in the Klorvess Liquid on slide with No. 0 fine, soft fur pen, often
Temperature is lower to be placed 30 minutes, then Klorvess Liquid is siphoned away or is transferred to spermatophore with fine, soft fur pen new with liquid-transfering gun or rubber head dropper
On slide, spermatophore is gently staved with No. 0 insect needle or thinner needle-shaped tool, and it is quick new toward the drop of instillation one on slide
The fixer of configuration（Methanol：Glacial acetic acid=3:1）, 30 minutes are fixed, at least 3 fixers are added dropwise in supplement in the process（Because
Fixer evaporation rate is fast）, after liquid to be fixed is dried, toward slide on instill Giemsa stain（1 part of stoste, PH7.2 phosphoric acid delays
9 parts of dilutions of fliud flushing）, dye 60 minutes under normal temperature, finally rinsed out dye liquor with clear water, through spontaneously drying or being dried up with hair-dryer
Afterwards, toward instilling an excellent Ba Laer of drop above the position that color is caught on slide, then covered, and with marking pen to dye
The position of colouring carries out circle note to facilitate observation, and i.e. observable Tianjin river amblyseius as predatory mite is male under an optical microscope after sample fixes
Mite sperm.
Claims (1)
1. the preparation method of Tianjin river amblyseius as predatory mite hero mite sperm slide sample, it is characterized in that male from Tianjin river amblyseius as predatory mite to mate
The method that sperm slide sample is made after spermatophore is obtained on mite gnathosoma, comprising the acquisition of male and female mite in mating, the sizing of male and female mite,
The separation of male and female mite, the acquisition of spermatophore, the acquisition of sperm and slide sample prepare 5 parts, and all operations are except the male and female in mating
Mite sizing is outer to be carried out under the microscope, and concrete operations are：
（1）The acquisition of male and female mite in mating：Out of Tianjin river amblyseius as predatory mite raising population, the ovum for choosing Tianjin river amblyseius as predatory mite is individually raised extremely
Male and female mite pairing is carried out after into mite, 10 minutes or so male and female mites of pairing can mate, so as to male and female mite in being mated；
（2）The sizing of male and female mite in mating：Within Tianjin river amblyseius as predatory mite post-coitum 0-120 minutes, it will be in mating process
Male and female individual is positioned over 3-5 minutes in -80 DEG C of ultra low temperature freezers or liquid nitrogen together with breeding apparatus, carries out male and female mite sizing；；
（3）The separation of male and female mite after sizing：Male and female mite is taken out together with breeding apparatus after sizing, then with fine, soft fur pen from raising
The male and female mite of mating is chosen on slide in device, recycles the instrument such as No. 0 insect needle and fine, soft fur pen by the male and female mite of mating
Gently separate, prevent from destroying spermatophore；
（4）The acquisition of spermatophore：After male and female mite successfully separates, Hubei Province body of male mite is scaled off using scalpel, retains gnathosoma, in jaw
The spermatophore in spermatodactyl top is readily observed on body, then will be put into 70%-100% under the light picking of spermatophore with fine, soft fur nib
Preserve in alcohol or directly utilize；
（5）It is prepared by the acquisition of sperm and slide sample：The Klorvess Liquid of a drop 0.075mol/L is instilled in load with rubber head dropper
The middle position of slide, then Tianjin river amblyseius as predatory mite spermatophore is positioned in the Klorvess Liquid on slide with No. 0 fine, soft fur pen, often
Temperature is lower to be placed 30 minutes, then Klorvess Liquid is siphoned away or is transferred to spermatophore with fine, soft fur pen new with liquid-transfering gun or rubber head dropper
On slide, spermatophore is gently staved with No. 0 insect needle or thinner needle-shaped tool, and it is quick new toward the drop of instillation one on slide
The fixer of configuration（Methanol：Glacial acetic acid=3:1）, 30 minutes are fixed, at least 3 fixers are added dropwise in supplement in the process（Because
Fixer evaporation rate is fast）, after liquid to be fixed is dried, toward slide on instill Giemsa stain（1 part of stoste, the phosphorus of PH=7.2
9 parts of dilutions of acid buffer）, dye 60 minutes under normal temperature, finally rinsed out dye liquor with clear water, through spontaneously drying or using hair-dryer
After drying, toward slide on caught an excellent Ba Laer of drop, then covered, and use marking pen instilled above the position of color
Circle note is carried out to the position for catching color to facilitate observation, i.e. observable Tianjin river is blunt under an optical microscope after sample fixes pacifies
Mite hero mite sperm.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6058878A (en) * | 1998-06-17 | 2000-05-09 | California Polytechnic State University Foundation | Protozoan free colonies of lepidoptera |
| CN102607907A (en) * | 2012-02-24 | 2012-07-25 | 东北农业大学 | Paraffin section method for fern gametophytes |
| CN103868768A (en) * | 2014-02-14 | 2014-06-18 | 河南省农业科学院植物保护研究所 | Treatment method of scanning electron microscope samples of insect tentacles and appendages |
| CN105486554A (en) * | 2015-11-19 | 2016-04-13 | 新疆农业大学 | Formula and method for immobilizing tick sample for scanning electron microscopy |
-
2017
- 2017-09-09 CN CN201710808918.2A patent/CN107568199A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6058878A (en) * | 1998-06-17 | 2000-05-09 | California Polytechnic State University Foundation | Protozoan free colonies of lepidoptera |
| CN102607907A (en) * | 2012-02-24 | 2012-07-25 | 东北农业大学 | Paraffin section method for fern gametophytes |
| CN103868768A (en) * | 2014-02-14 | 2014-06-18 | 河南省农业科学院植物保护研究所 | Treatment method of scanning electron microscope samples of insect tentacles and appendages |
| CN105486554A (en) * | 2015-11-19 | 2016-04-13 | 新疆农业大学 | Formula and method for immobilizing tick sample for scanning electron microscopy |
Non-Patent Citations (1)
| Title |
|---|
| 李隆术等: "《蜱螨学》", 30 November 1988, 重庆出版社 * |
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Effective date of registration: 20180510 Address after: 271406 the village committee of Shi Zhuang village, Heshan Town, Ningyang County, Tai'an, Shandong Applicant after: Shandong first Bio Technology Co., Ltd. Address before: 230041, 4, garden 1, Internet garden, 17 Garden Avenue, Baohe Economic Development Zone, Hefei, Anhui. Applicant before: Hefei Zhong Bao Hui Agricultural Biotechnology Co., Ltd. |
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Application publication date: 20180112 |
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