CN107643197A - A kind of Predatory Mites that prepare are cut open the belly the method for ovum chromosome - Google Patents
A kind of Predatory Mites that prepare are cut open the belly the method for ovum chromosome Download PDFInfo
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- CN107643197A CN107643197A CN201710808930.3A CN201710808930A CN107643197A CN 107643197 A CN107643197 A CN 107643197A CN 201710808930 A CN201710808930 A CN 201710808930A CN 107643197 A CN107643197 A CN 107643197A
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- belly
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Abstract
The present invention relates to a kind of Predatory Mites that prepare to cut open the belly the method for ovum chromosome, it is characterized in that artificially female into the method that not yet output ovum is obtained in mite body and ovum chromosome of cutting open the belly is prepared on slide from Predatory Mites using the form cut open the belly, comprising the female acquisition into mite of Predatory Mites to be cut open the belly and its placement on slide, Predatory Mites cut open the belly ovum acquisition deposited with it, ovum chromosome slide sample of cutting open the belly 5 steps of preparation.The present invention, which very can cleverly obtain Predatory Mites and cut open the belly, ovum and prepares chromosome, the preparation ovum chromosome that can be cut open the belly for observation Predatory Mites of ovum chromosome of cutting open the belly is monoploid or diploid and heterogeneousization whether occurs to provide raw material, the cut open the belly acquisition of ovum chromosome of different time sections is advantageous to find the time node that chromosome heterogeneousization occurs earliest, can provide strong evidence to the reproduction mechanism for disclosing Predatory Mites.
Description
Technical field
The present invention relates to a kind of Predatory Mites that prepare to cut open the belly the method for ovum chromosome, belongs to Predatory Mites fundamental biological knowledge research neck
Domain.
Background technology
Predatory Mites are important small-sized pest natural enemies, and some Predatory Mites reproductions have particularity, i.e. male and female offspring is equal
Come from development of fertilized ova, but male offspring develops into class monoploid because being disappeared after paternal gene heterochromatinization.This is recognized
To be determined by some Predatory Mites such as special mode of reproduction of Phytoseiidae Predatory Mites, i.e., false arrhenotocia
(Pseudoarrhenotoky) (Hoy, 1985; Nagalkerke & Sabelis 1998; Sabelis et al.
2002).In recent years, Predatory Mites are in addition to as biological control natural enemy, and the research of its biological habit also attracts attention, especially
This special reproduction phenomenon of false arrhenotocia.
Nelson-Rees et al.（1980）Observe west it is quiet walk mite egg development during caryogram change think：
After ovum output in 0-24 hours, the embryonic cell half chromatin of part ovum（3/6）Beginning pyknosis is heterochromatin；After output
In 24-48 hours, there is heterochromatinization phenomenon in most cells in this partial embryonic, and subsequent heterochromatin is discharged cell,
Embryonic development is monoploid male.This result of study be still at present delivered it is most detailed to Phytoseiidae Sex Determination theory
Description, it thinks that initial stage after ovum output occurs for phytoseiid mite Sex Determination.
It is true whether such, heterochromatinization start from actually when, in order to understand fully the ovum of Predatory Mites before output whether
Monoploid caryogram be present, i.e., before ovum output development for male mite ovum in a hemichromosome whether have occurred and that heterogeneousization or
Lose, it is necessary to study ovum chromosome of cutting open the belly, observation chromosome is monoploid or diploid and whether heterogeneousization occurs.
Because Predatory Mites individual is small, only hundreds of microns, and be easy in the more fragile operating process of body to it
Producing damage causes body fluid to outflow, and body trunk is flat can not to carry out any experiment, and progress micromanipulation difficulty is quite big, is difficult to from list
One individual obtains complete tissue samples, therefore either from morphosis to cellular level, research chromosome be monoploid or
Diploid and that heterogeneousization whether occurs is all very difficult.In addition, research Predatory Mites ovum chromosome of cutting open the belly is monoploid or two
Times body and whether heterogeneousization occurs and cut open the belly ovum slide sample, it is necessary to make and observe the ovum of cutting open the belly in slide sample under the microscope
Chromosome, however in available data how to obtain or make Predatory Mites cut open the belly ovum chromosome method report.
Being cut open the belly for observation Predatory Mites, ovum chromosome is monoploid or diploid and heterogeneousization whether occurs, and the present invention provides one
Predatory Mites kind are prepared to cut open the belly the method for ovum chromosome.
The content of the invention
It is an object of the invention to provide a kind of Predatory Mites that prepare to cut open the belly the method for ovum chromosome, can be to obtain Predatory Mites cut open
Cut open the belly ovum chromosome, observation Predatory Mites ovum chromosome of cutting open the belly of abdomen ovum and preparation is monoploid or diploid and whether heterogeneousization occurs
Concrete operation method is provided.
The present invention is artificially female into obtaining not yet output ovum and in slide in mite body from Predatory Mites using the form cut open the belly
Upper preparation is cut open the belly the method for ovum chromosome, comprising the female acquisition into mite of Predatory Mites to be cut open the belly and its placement on slide, is caught
Food mite cut open the belly ovum acquisition deposited with it, chromosome slide sample prepare 5 steps, in addition to chromosome observe, it is all operate
Carried out under Stereo microscope, concrete operation method is：
（1）The female acquisition into mite of Predatory Mites to be cut open the belly：From Predatory Mites raising colony in picking Predatory Mites ovum individually raising to
Into mite, a female hero is then placed on the small interior of raising and matched, the male and female Predatory Mites just sprouted wings can be mated quickly, be treated
After the completion of mating, continue to provide sufficient prey into mite to female, 12-15 hours are female to expand to have to obtain after ovum into mite belly and wait to cut open
Abdomen Predatory Mites are female into mite；
（2）The female placement into mite on slide of Predatory Mites to be cut open the belly：Clear water is dripped to slide, to be dissected female of picking
Be placed on into mite in the clear water on slide, and with insect needle and fine, soft fur pen keep it is female into the mite back side upward, clear water infusion volume should not
Excessively, avoid female swimming on water layer into mite, it is ensured that female to remain able to touch slide into mite；
（3）Predatory Mites are cut open the belly the acquisition of ovum：The female Hubei Province body portion into mite of Predatory Mites first is pinned with No. 3 thick insect needles, then it is thin with No. 0
Insect needle ventrad since Predatory Mites head, which is gently extruded to belly, to be ruptured, you can sees that Predatory Mites are cut open the belly ovum, then with No. 0
Fine, soft fur nib portion gently pushes the tissue around ovum aside, and this step must be carried out in clear water, prevents that ovum dehydration is shrivelled；
（4）Predatory Mites are cut open the belly the storage of ovum：Predatory Mites ovum of cutting open the belly is in overlong time in water and is easily broken, it is necessary to quickly with No. 0
Fine, soft fur pen, which is positioned over after ovum of cutting open the belly carefully is chosen from water in 100% alcohol, to be deposited standby, that is, is obtained Predatory Mites and cut open the belly ovum；
（5）It is prepared by chromosome slide sample：The Klorvess Liquid of a drop 0.075mol/L is instilled in slide with rubber head dropper
Middle position, then the Predatory Mites of taking-up ovum of cutting open the belly is positioned in the Klorvess Liquid on slide with No. 0 fine, soft fur pen, normal temperature
It is lower to place 30 minutes, then with liquid-transfering gun or rubber head dropper by Klorvess Liquid siphon away or will cut open the belly ovum be transferred to fine, soft fur pen it is new
On slide, gently staved, and quick dripped toward instilling one on slide with No. 0 insect needle or thinner the needle-shaped tool ovum that will cut open the belly
The fixer newly configured（Methanol：Glacial acetic acid=3:1）, 30 minutes are fixed, at least 3 fixers are added dropwise in supplement in the process（Cause
It is fast for fixer evaporation rate）, after liquid to be fixed is dried, toward slide on instill Giemsa stain（1 part of stoste, PH7.2 phosphoric acid
9 parts of dilutions of buffer solution）, dye 60 minutes under normal temperature, finally rinsed out dye liquor with clear water, through spontaneously drying or being blown with hair-dryer
After dry, toward slide on caught and instill one above the position of color and drip excellent Ba Laer, then covered, and with marking pen pair
The position for catching color carries out circle note to facilitate observation, and i.e. observable Predatory Mites are cut open the belly under an optical microscope after sample fixes
Ovum chromosome.
The Predatory Mites that the present invention very can cleverly obtain different time sections are cut open the belly and ovum and prepare chromosome, ovum of cutting open the belly dye
The preparation of the colour solid ovum chromosome that can be cut open the belly for observation Predatory Mites is monoploid or diploid and heterogeneousization whether occurs to provide original
Material, the cut open the belly acquisition of ovum chromosome of different time sections are advantageous to find the time node that chromosome heterogeneousization occurs earliest, this
Strong evidence can be provided to the reproduction mechanism for disclosing Predatory Mites.
Embodiment
Predatory Mites of the embodiment by taking Amblyseius persidolongispinosus as an example are cut open the belly ovum chromosome specimen preparation method
Concrete operation method includes following 5 part, and in addition to chromosome is observed, all operations are carried out under Stereo microscope：
（1）The female acquisition into mite of Amblyseius persidolongispinosus to be cut open the belly：Picking plan becomes mildewed blunt from the raising colony of Amblyseius persidolongispinosus
Individually a female hero extremely into mite, is then placed on the small interior of raising and matched the ovum of peaceful mite by raising, the male and female plan length just sprouted wings
Hair amblyseius as predatory mite can be mated quickly, be waited after the completion of mating, and continue to provide sufficient prey into mite to female, 12-15 hours it is female into
Mite belly, which expands, that to obtain Amblyseius persidolongispinosus to be cut open the belly after ovum female into mite；
（2）The female placement into mite on slide of Amblyseius persidolongispinosus to be cut open the belly：Clear water is dripped to slide, and picking is waited to solve
Cut open it is female be placed on into mite in the clear water on slide, and with insect needle and fine, soft fur pen keep it is female into the mite back side upward, clear water instills
Amount should not be excessive, avoids female swimming on water layer into mite, it is ensured that female to remain able to touch slide into mite；
（3）Amblyseius persidolongispinosus is cut open the belly the acquisition of ovum：First the female Hubei Province body portion into mite of Amblyseius persidolongispinosus is pinned with No. 3 thick insect needles
Point, then gently extruded to belly and ruptured with No. 0 thin insect needle ventrad since Amblyseius persidolongispinosus head, you can see plan length
Hair amblyseius as predatory mite is cut open the belly ovum, is then gently pushed the tissue around ovum aside with No. 0 fine, soft fur nib portion, this step must be in clear water
Carry out, prevent that ovum dehydration is shrivelled；
（4）Amblyseius persidolongispinosus is cut open the belly the storage of ovum：Amblyseius persidolongispinosus ovum of cutting open the belly is in overlong time in water and is easily broken, and needs
It is positioned over after quickly ovum of cutting open the belly carefully to be chosen from water with No. 0 fine, soft fur pen in 100% alcohol and deposits standby, that is, is intended
The amblyseius as predatory mite that becomes mildewed is cut open the belly ovum；
（5）It is prepared by chromosome slide sample：The Klorvess Liquid of a drop 0.075mol/L is instilled in slide with rubber head dropper
Middle position, then the Amblyseius persidolongispinosus of taking-up is cut open the belly the Klorvess Liquid that ovum is positioned on slide with No. 0 fine, soft fur pen
In, placed 30 minutes under normal temperature, then Klorvess Liquid is siphoned away or will cut open the belly ovum fine, soft fur pen with liquid-transfering gun or rubber head dropper and is turned
Move on on new slide, gently staved with No. 0 insect needle or thinner the needle-shaped tool ovum that will cut open the belly, and it is quick toward on slide
Instill the fixer that a drop newly configures（Methanol：Glacial acetic acid=3:1）, 30 minutes are fixed, at least supplement is added dropwise 3 times in the process
Fixer（Because fixer evaporation rate is fast）, after liquid to be fixed is dried, toward slide on instill Giemsa stain（1 part of stoste,
The 9 parts of dilutions of PH7.2 phosphate buffers）, dye 60 minutes under normal temperature, finally rinsed out dye liquor with clear water, through spontaneously drying or
After being dried up with hair-dryer, toward slide on caught an excellent Ba Laer of drop instilled above the position of color, then covered, and
Circle note is carried out to the position for catching color with marking pen to facilitate observation, the i.e. observable under an optical microscope after sample fixes
Amblyseius persidolongispinosus is cut open the belly ovum chromosome.
Claims (1)
1. a kind of Predatory Mites that prepare are cut open the belly the method for ovum chromosome, it is characterized in that using the form cut open the belly artificially from Predatory Mites it is female into
The method that not yet output ovum is obtained in mite body and ovum chromosome of cutting open the belly is prepared on slide, it is female into mite comprising Predatory Mites to be cut open the belly
Acquisition with its placement on slide, Predatory Mites cut open the belly ovum acquisition and its deposit, ovum chromosome slide sample system of cutting open the belly
Standby 5 steps, in addition to chromosome is observed, all operations are carried out under Stereo microscope, and concrete operation method is：
（1）The female acquisition into mite of Predatory Mites to be cut open the belly：From Predatory Mites raising colony in picking Predatory Mites ovum individually raising to
Into mite, a female hero is then placed on the small interior of raising and matched, the male and female Predatory Mites just sprouted wings can be mated quickly, be treated
After the completion of mating, continue to provide sufficient prey into mite to female, 12-15 hours are female to expand to have to obtain after ovum into mite belly and wait to cut open
Abdomen Predatory Mites are female into mite；
（2）The female placement into mite on slide of Predatory Mites to be cut open the belly：Clear water is dripped to slide, to be dissected female of picking
Be placed on into mite in the clear water on slide, and with insect needle and fine, soft fur pen keep it is female into the mite back side upward, clear water infusion volume should not
Excessively, avoid female swimming on water layer into mite, it is ensured that female to remain able to touch slide into mite；
（3）Predatory Mites are cut open the belly the acquisition of ovum：The female Hubei Province body portion into mite of Predatory Mites first is pinned with No. 3 thick insect needles, then it is thin with No. 0
Insect needle ventrad since Predatory Mites head, which is gently extruded to belly, to be ruptured, you can sees that Predatory Mites are cut open the belly ovum, then with No. 0
Fine, soft fur nib portion gently pushes the tissue around ovum aside, and this step must be carried out in clear water, prevents that ovum dehydration is shrivelled；
（4）Predatory Mites are cut open the belly the storage of ovum：Predatory Mites ovum of cutting open the belly is in overlong time in water and is easily broken, it is necessary to quickly with No. 0
Fine, soft fur pen, which is positioned over after ovum of cutting open the belly carefully is chosen from water in 100% alcohol, to be deposited standby, that is, is obtained Predatory Mites and cut open the belly ovum；
（5）It is prepared by ovum chromosome slide sample of cutting open the belly：The Klorvess Liquid of a drop 0.075mol/L is instilled in load with rubber head dropper
The middle position of slide, then the Predatory Mites of taking-up are cut open the belly the Klorvess Liquid that ovum is positioned on slide with No. 0 fine, soft fur pen
In, placed 30 minutes under normal temperature, then Klorvess Liquid is siphoned away or will cut open the belly ovum fine, soft fur pen with liquid-transfering gun or rubber head dropper and is turned
Move on on new slide, gently staved with No. 0 insect needle or thinner the needle-shaped tool ovum that will cut open the belly, and it is quick toward on slide
Instill the fixer that a drop newly configures（Methanol：Glacial acetic acid=3:1）, 30 minutes are fixed, at least supplement is added dropwise 3 times in the process
Fixer（Because fixer evaporation rate is fast）, after liquid to be fixed is dried, toward slide on instill Giemsa stain（1 part of stoste,
The 9 parts of dilutions of PH7.2 phosphate buffers）, dye 60 minutes under normal temperature, finally rinsed out dye liquor with clear water, through spontaneously drying or
After being dried up with hair-dryer, toward slide on caught an excellent Ba Laer of drop instilled above the position of color, then covered, and
Circle note is carried out to the position for catching color with marking pen to facilitate observation, the i.e. observable under an optical microscope after sample fixes
Predatory Mites are cut open the belly ovum chromosome.
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Citations (5)
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|---|---|---|---|---|
| US6058878A (en) * | 1998-06-17 | 2000-05-09 | California Polytechnic State University Foundation | Protozoan free colonies of lepidoptera |
| CN101822239A (en) * | 2010-05-07 | 2010-09-08 | 华南农业大学 | Device and method for collecting mite eggs |
| 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 CN201710808930.3A patent/CN107643197A/en active Pending
Patent Citations (5)
| 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 |
| CN101822239A (en) * | 2010-05-07 | 2010-09-08 | 华南农业大学 | Device and method for collecting mite eggs |
| 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 (3)
| Title |
|---|
| 李隆术等: "《蜱螨学》", 30 November 1988 * |
| 陈春生等: "《上海真厉螨的染色体组型及其C带研究》", 《遗传学报》 * |
| 陈春生等: "《茅舍血厉螨核型及染色体的C带、G带的研究", 《动物学研究》 * |
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Application publication date: 20180130 |