CN106973864A - A kind of breeding apparatus and its application method suitable for white backed planthopper injection RNA interference experiments - Google Patents
A kind of breeding apparatus and its application method suitable for white backed planthopper injection RNA interference experiments Download PDFInfo
- Publication number
- CN106973864A CN106973864A CN201710278674.1A CN201710278674A CN106973864A CN 106973864 A CN106973864 A CN 106973864A CN 201710278674 A CN201710278674 A CN 201710278674A CN 106973864 A CN106973864 A CN 106973864A
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- China
- Prior art keywords
- backed planthopper
- white backed
- door body
- transparent plastic
- gene
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- 241000176086 Sogatella furcifera Species 0.000 title claims abstract description 43
- 238000009395 breeding Methods 0.000 title claims abstract description 18
- 230000001488 breeding effect Effects 0.000 title claims abstract description 18
- 230000009368 gene silencing by RNA Effects 0.000 title claims abstract description 17
- 238000012228 RNA interference-mediated gene silencing Methods 0.000 title claims abstract description 14
- 238000002347 injection Methods 0.000 title claims abstract description 14
- 239000007924 injection Substances 0.000 title claims abstract description 14
- 238000002474 experimental method Methods 0.000 title claims abstract description 13
- 238000000034 method Methods 0.000 title claims abstract description 10
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 34
- 235000007164 Oryza sativa Nutrition 0.000 claims abstract description 24
- 235000009566 rice Nutrition 0.000 claims abstract description 24
- 229920003023 plastic Polymers 0.000 claims abstract description 20
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims abstract description 14
- 102000040650 (ribonucleotides)n+m Human genes 0.000 claims abstract description 11
- 238000005070 sampling Methods 0.000 claims abstract description 10
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 7
- 229910052799 carbon Inorganic materials 0.000 claims abstract description 7
- 241000209094 Oryza Species 0.000 claims description 23
- 108010057988 ecdysone receptor Proteins 0.000 claims description 4
- 238000009434 installation Methods 0.000 claims description 4
- 229920002101 Chitin Polymers 0.000 claims description 3
- 108090000364 Ligases Proteins 0.000 claims description 3
- 108010087472 Trehalase Proteins 0.000 claims description 3
- 238000001764 infiltration Methods 0.000 claims description 3
- 230000008595 infiltration Effects 0.000 claims description 3
- 238000003780 insertion Methods 0.000 claims description 3
- 230000037431 insertion Effects 0.000 claims description 3
- 235000015097 nutrients Nutrition 0.000 claims description 3
- 239000000243 solution Substances 0.000 claims description 3
- 101150067597 treh gene Proteins 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 101150095206 akirin gene Proteins 0.000 claims 1
- 238000011160 research Methods 0.000 abstract description 8
- 240000007594 Oryza sativa Species 0.000 abstract 1
- 229920000742 Cotton Polymers 0.000 description 9
- 238000005516 engineering process Methods 0.000 description 5
- 235000016709 nutrition Nutrition 0.000 description 4
- 230000035764 nutrition Effects 0.000 description 4
- 108091030071 RNAI Proteins 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 101710204790 Akirin Proteins 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 235000013339 cereals Nutrition 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- 241001466044 Delphacidae Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 108700001097 Insect Genes Proteins 0.000 description 1
- 206010033799 Paralysis Diseases 0.000 description 1
- 241000517324 Pediculidae Species 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000003651 drinking water Substances 0.000 description 1
- 235000020188 drinking water Nutrition 0.000 description 1
- 239000004744 fabric Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 230000030279 gene silencing Effects 0.000 description 1
- 238000012226 gene silencing method Methods 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000007654 immersion Methods 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 208000028454 lice infestation Diseases 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000007639 printing Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000012857 repacking Methods 0.000 description 1
- 238000009738 saturating Methods 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 239000002341 toxic gas Substances 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- 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
- A01K67/0333—Genetically modified invertebrates, e.g. transgenic, polyploid
- A01K67/0337—Genetically modified Arthropods
- A01K67/0339—Genetically modified insects, e.g. Drosophila melanogaster, medfly
-
- 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
- A01K2207/00—Modified animals
- A01K2207/05—Animals modified by non-integrating nucleic acids, e.g. antisense, RNAi, morpholino, episomal vector, for non-therapeutic purpose
-
- 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
- A01K2227/00—Animals characterised by species
- A01K2227/70—Invertebrates
- A01K2227/706—Insects, e.g. Drosophila melanogaster, medfly
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
This application discloses the breeding apparatus suitable for white backed planthopper injection RNA interference experiments, including transparent plastic bottle and for cultivating the hypothallus of paddy rice, the hypothallus is located at the bottom in transparent plastic bottle bottle, the body of the transparent plastic bottle is provided with opening, the door body that can prevent that white backed planthopper from passing in and out is hinged with the body at the opening, the door body offers provided with active carbon layer and the magnetic lock used cooperatively with opening, the door body and is hinged with sampling gate on sample tap, sample tap.In use, opening door body, the white backed planthopper for having injected specific gene dsRNA is put into transparent plastic bottle at door body, door body is closed；Cultivate after 24h, observe the change of white backed planthopper phenotype, observed, taken 4 ~ 5 times by sample tap insect-taking every 24h, record experimental result.Present apparatus structures and methods set reasonable, are more suitable for utilizing the research for injecting RNA perturbation techniques for white backed planthopper gene function.
Description
Technical field
The present invention relates to technical field of bioengineering, and in particular to paddy rice important pests white backed planthopper gene functional research is auxiliary
Technical field of tools is helped, what is further related to is a kind of breeding apparatus suitable for white backed planthopper injection RNA interference experiments and its makes
Use method.
Background technology
Paddy rice is the important cereal crops of China, and yield ranks first in various cereal crops middle positions.White backed planthopper is subordinate to together
Wing mesh, Delphacidae insect, not only thorn absorption drinking water rice is so that the normal growth of hazard rice, can more propagate plant virus, cause
Paddy rice strain paralysis lodging, the paddy rice underproduction is even had no harvest.It is gene level research each gene of white backed planthopper from the essence of white backed planthopper
Function will be helpful to the research and preventing and treating of this insect.RNA is disturbed（RNA interference, are abbreviated as RNAi）Refer to one
Plant by double-stranded RNA（Double-stranded RNA, dsRNA）The gene silencing phenomenon of induction.Due to can be with using RNAi technology
Specific depletion or the expression for closing specific gene, so the technology is widely used to explore the function of gene.
Since RNAi technology self-discovery, in the research for being widely used to insect genes function, but for white backed planthopper
Gene functional research technology but have no report.
The content of the invention
After the problem of present invention lacks the gene function Prevention Technique of white backed planthopper for prior art, is there is provided one kind injection
It is easy to the breeding apparatus observed and sampled, the research and preventing and treating for white backed planthopper gene function provide facility.
Concrete scheme is：A kind of breeding apparatus suitable for white backed planthopper injection RNA interference experiments, including transparent plastic
Bottle and the hypothallus for cultivating paddy rice, the hypothallus are located at the bottom in transparent plastic bottle bottle, the transparent plastic bottle
Body is provided with opening, and the door body that can prevent that white backed planthopper from passing in and out, the door body are hinged with the body at the opening
Provided with active carbon layer and the magnetic lock used cooperatively with opening, the door body is offered to be hinged with sample tap, sample tap and taken
Sample door.
Further, the top of the transparent plastic bottle offers and is arranged with gauze on bottleneck, the bottleneck.Bottleneck and yarn
The purpose of cloth is to improve the air permeable effect of device.
A kind of application method of breeding apparatus suitable for white backed planthopper injection RNA interference experiments, comprises the following steps：
Step 1: the installation of breeding apparatus：Door body is opened, hypothallus bottom of bottle is laid on, lower floor's infiltration paddy rice nutrient solution will
Band root Seedling Stage paddy rice cleans up, and dries in water stain rear insertion hypothallus, upright and fixed；
Step 2: the dispensing of white backed planthopper：The white backed planthopper for having injected specific gene dsRNA is put into transparent plastic at door body
In bottle, door body is closed；
Step 3: sampling：Cultivate after 24h, observe the change of white backed planthopper phenotype, observed, taken by sample tap insect-taking every 24h
4 ~ 5 times, record experimental result.
Further, the specific gene dsRNA is the double-stranded RNA of white backed planthopper specific gene, the specific gene bag
Include immunogene, grow related gene or other functional genes, the related gene that grows includes moulting hormone
Acceptor gene EcR, chitin synthetase gene Chs and trehalase gene Treh, the immunogene include Toll genes and
Akirin genes.
The operation principle and beneficial effect of the present invention：For the device of white backed planthopper injection RNA interference experiments, using saturating
Bright plastic bottle easy processing repacking, while the characteristics of printing opacity is conveniently observed and do not influence Rice Photosynthesis, opens up on body
Opening, opening is hinged door body, and door body is provided with active carbon layer and magnetic lock, and active carbon layer is arranged on the environment in bottle in door body
With external environment condition opposing spaced apart, air is entered back into bottle after being filtered through active carbon layer, air is become pure, it is to avoid after pollution
The growth of air Whitebacked Planthopper is impacted, in the case where running into special circumstances, such as fire or toxic gas leakage,
The device of the present invention may insure that the white backed planthopper in bottle is not affected still in 10 ~ 20min clocks, be provided to experimenter
The time of redemption, it is to avoid the waste of previous work；It is connected and locks with opening by magnetic lock, gently one draws one to push away and can open
Close, operation more facilitates, while the more conducively installation of paddy rice and hypothallus, hypothallus is cotton layer or soil horizon, paddy rice
In the presence of rice nutrition liquid and illumination, grown in breeding apparatus, food is provided to white backed planthopper；It will inject special again
Gene dsRNA white backed planthopper is delivered into breeding apparatus, during culture, utilizes the sample tap dialogue backward flight in door body
Lice is sampled, it is to avoid directly by opening during sampling, and it is excessive to be open, and the white backed planthopper device that flies out influences test effect,
It can also be operated during supplement rice nutrition liquid by sample tap；Each part of the present apparatus is readily obtained, with simple in construction and warp
The characteristics of Ji is practical, is more suitable for utilizing RNA perturbation techniques to be used for the research of white backed planthopper gene function.
Brief description of the drawings
Fig. 1 is structural representation of the present invention suitable for the breeding apparatus of white backed planthopper injection RNA interference experiments.
Embodiment
Below by embodiment, the present invention is further detailed explanation：
Illustrate the embodiment of the present invention, but protection scope of the present invention by example below, be not limited to this.
Reference：Gauze 1, bottleneck 2, body 3, paddy rice 4, sampling gate 5, sample tap 6, door body 7, cotton layer 8.
Embodiment 1 is as shown in figure 1, the breeding apparatus suitable for white backed planthopper injection RNA interference experiments of the present embodiment
Including a transparent square transparent plastic bottle, gauze 1 is covered on bottleneck 2, fixed with rubber band, the side of body 3 is offered
Fine and closely woven wire netting is hinged with body 3 at opening, opening to be made in door body 7, door body 7 covered with active carbon layer, and can be outside
Open, door body 7 is provided with magnetic lock, is used cooperatively with opening, for quick locking door body 7, sampling is further opened with door body 7
Mouth 6, sample tap 6 can accommodate stretching into for test tube, for insect-taking, and same be made up of fine and closely woven wire netting is hinged with sample tap 6 and is taken
Sample door 5；Cotton layer 8 is provided with inside transparent plastic bottle, the bottom of cotton layer 8 is moistened by rice nutrition immersion, and paddy rice 4 is fixed on vertically
In cotton layer 8.
In use, comprising the following steps：Step 1: the installation of breeding apparatus：Door body 7 is opened, cotton layer 8 is laid on
Bottom of bottle, lower floor's infiltration paddy rice nutrient solution of cotton layer 8, will be cleaned up with root Seedling Stage paddy rice 4, dry water stain rear insertion cotton
It is upright and fixed in layer 8；
Step 2: the dispensing of white backed planthopper：The white backed planthopper for having injected Toll genes is put into transparent plastic bottle at door body 7
In, close door body 7；
Step 3: sampling：Cultivate after 24h, observe the change of white backed planthopper phenotype, sampling gate 5 is opened every 24h, pass through sampling
Mouth 6 insect-takings observation, takes 4 ~ 5 times, records experimental result.
When culture, by sample tap 6, rice nutrition liquid is supplemented in cotton layer 8.
Square transparent plastic bottle is substituted for circular non-opaque plastic bottle by embodiment 2 with differing only in for embodiment 1, will
Toll genes are substituted for Akirin genes.
Toll genes are substituted for ecdysone receptor gene EcR by embodiment 3 with differing only in for embodiment 1.
Toll genes are substituted for chitin synthetase gene Chs by embodiment 4 with differing only in for embodiment 1.
Toll genes are substituted for trehalase gene Treh by embodiment 5 with differing only in for embodiment 1.
Above-described is only that the known general knowledge such as concrete structure and characteristic is not made herein in embodiments of the invention, scheme
Excessive description., without departing from the structure of the invention, can be with it should be pointed out that for those skilled in the art
Several modifications and improvements are made, these should also be considered as protection scope of the present invention, these are implemented all without the influence present invention
Effect and practical applicability.The scope of protection required by this application should be based on the content of the claims, in specification
Embodiment etc. records the content that can be used for explaining claim.
Claims (4)
1. a kind of breeding apparatus suitable for white backed planthopper injection RNA interference experiments, it is characterised in that including transparent plastic bottle
With the hypothallus for cultivating paddy rice, the hypothallus is located at the bottom in transparent plastic bottle bottle, the bottle of the transparent plastic bottle
Opening is provided with it, is hinged with the body at the opening in the door body that can prevent that white backed planthopper from passing in and out, the door body
The magnetic lock used cooperatively provided with active carbon layer and with opening, the door body, which is offered, is hinged with sampling on sample tap, sample tap
Door.
2. the breeding apparatus according to claim 1 suitable for white backed planthopper injection RNA interference experiments, its feature exists
In the top of the transparent plastic bottle is offered is arranged with gauze on bottleneck, the bottleneck.
3. the application method of the breeding apparatus according to claim 2 suitable for white backed planthopper injection RNA interference experiments,
It is characterised in that it includes following steps：
Step 1: the installation of breeding apparatus：Door body is opened, hypothallus bottom of bottle is laid on, lower floor's infiltration paddy rice nutrient solution will
Band root Seedling Stage paddy rice cleans up, and dries in water stain rear insertion hypothallus, upright and fixed；
Step 2: the dispensing of white backed planthopper：The white backed planthopper for having injected specific gene dsRNA is put into transparent plastic at door body
In bottle, door body is closed；
Step 3: sampling：Cultivate after 24h, observe the change of white backed planthopper phenotype, observed, taken by sample tap insect-taking every 24h
4 ~ 5 times, record experimental result.
4. the application method of the breeding apparatus according to claim 3 suitable for white backed planthopper injection RNA interference experiments,
Characterized in that, the specific gene dsRNA is the double-stranded RNA of white backed planthopper specific gene, the specific gene includes immune
Gene, grow related gene or other functional genes, and the related gene that grows includes ecdysone receptor base
Because of EcR, chitin synthetase gene Chs and trehalase gene Treh, the immunogene includes Toll genes and Akirin
Gene.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710278674.1A CN106973864A (en) | 2017-04-25 | 2017-04-25 | A kind of breeding apparatus and its application method suitable for white backed planthopper injection RNA interference experiments |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710278674.1A CN106973864A (en) | 2017-04-25 | 2017-04-25 | A kind of breeding apparatus and its application method suitable for white backed planthopper injection RNA interference experiments |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106973864A true CN106973864A (en) | 2017-07-25 |
Family
ID=59344690
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710278674.1A Pending CN106973864A (en) | 2017-04-25 | 2017-04-25 | A kind of breeding apparatus and its application method suitable for white backed planthopper injection RNA interference experiments |
Country Status (1)
| Country | Link |
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| CN (1) | CN106973864A (en) |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020086356A1 (en) * | 2000-03-30 | 2002-07-04 | Whitehead Institute For Biomedical Research | RNA sequence-specific mediators of RNA interference |
| US20050080032A1 (en) * | 2003-07-02 | 2005-04-14 | Gross Paul S. | dsRNA induced specific and non-specific immunity in crustaceans and other invertebrates and biodelivery vehicles for use therein |
| CN102220341A (en) * | 2011-05-12 | 2011-10-19 | 南京农业大学 | Gene silencing technology-based lethal gene fragment Chitinase 7 of laodelphax striatellus and dsRNA (double-stranded RNA) thereof |
| CN103421775A (en) * | 2013-07-21 | 2013-12-04 | 浙江大学 | Nilaparvata lugen gene N14777 as well as encoded products and application thereof |
| CN103976162A (en) * | 2014-04-09 | 2014-08-13 | 东北农业大学 | Artificial feed for soybean pod borers, and preparation method and application thereof |
| CN104044912A (en) * | 2014-05-30 | 2014-09-17 | 楚天科技股份有限公司 | Bottle sampling testing device |
| CN204811555U (en) * | 2015-07-20 | 2015-12-02 | 中国水稻研究所 | Device is simply raised to planthopper short -term |
| CN205682214U (en) * | 2016-06-03 | 2016-11-16 | 贵州大学 | A kind of breeding apparatus of cacac moth |
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-
2017
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| US20020086356A1 (en) * | 2000-03-30 | 2002-07-04 | Whitehead Institute For Biomedical Research | RNA sequence-specific mediators of RNA interference |
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| CN102220341A (en) * | 2011-05-12 | 2011-10-19 | 南京农业大学 | Gene silencing technology-based lethal gene fragment Chitinase 7 of laodelphax striatellus and dsRNA (double-stranded RNA) thereof |
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| CN204811555U (en) * | 2015-07-20 | 2015-12-02 | 中国水稻研究所 | Device is simply raised to planthopper short -term |
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Application publication date: 20170725 |
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