CN106520768A - Breeding for brown planthopper population based on male sterilty after RNAi and control effect evaluation method - Google Patents
Breeding for brown planthopper population based on male sterilty after RNAi and control effect evaluation method Download PDFInfo
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- CN106520768A CN106520768A CN201610905214.2A CN201610905214A CN106520768A CN 106520768 A CN106520768 A CN 106520768A CN 201610905214 A CN201610905214 A CN 201610905214A CN 106520768 A CN106520768 A CN 106520768A
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/033—Rearing or breeding invertebrates; New breeds of invertebrates
- A01K67/0333—Genetically modified invertebrates, e.g. transgenic, polyploid
- A01K67/0335—Genetically modified worms
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- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/87—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; 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
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- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/14—Type of nucleic acid interfering N.A.
- C12N2310/141—MicroRNAs, miRNAs
Abstract
The invention relates to breeding for brown planthopper population based on male sterilty after RNAi and a control effect evaluation method. According to the invention, a RNAi breeding method is employed for silencing brown planthopper male testis growth gene PHF7, by detecting the mortality and gene silencing rate, the concentration of dsNlPHF7 is determined; the silenced male insect growth and reproduction parameter indexes are detected, which mainly comprise body weight, life span, accessory gland protein content and arginine content; the copulated male insect growth and reproduction parameter indexes are detected, which mainly comprise body weight, the life span, vitellogenin content, oviposition amount, and YLS richness; and the expression level of the vitellogenin gene in the copulated male insect fat is detected. The brown planthopper population increasing and controlling effects enable quantification evaluation by detecting the important reproduction parameter indexes of the brown planthopper population male insects and female insects, the method has the advantages of simple and fast operation, and has wide applicability, the method provides a novel control approach for preventing and treating brown planthopper, and has theory meaning and utility values.
Description
Technical field
The present invention relates to the indoor RNAi artificial diet techniques of China paddy rice important pests brown paddy plant hopper and quantitative assessment its control
The method of effect processed.
Background technology
Brown paddy plant hopper Nilaparvata lugensCategory Semiptera, Delphacidae is a kind of seasonality, migrating property, r-
Countermeasure, fulminant insect, widely distributed in Asia, South Asia, Southeast Asia, East Asia, South Pacific Ocean island and Australia have point
Cloth, also has to the east of Japan and some archipelagos in Southeast Asia, Micronesia and Papua New Guinea west from Pakistan
Distribution.Brown paddy plant hopper is monophagy insect, can only take food and raise up seed on paddy rice and on common wild rice.From 20th century 60
Since age, due to the popularization of Rice Dwarf kind, a large amount of change using N fertilizer and other cultivation steps, chemical insecticide
It is intensive the factor such as to use, promote the frequent population outbreak of brown paddy plant hopper, seriously threaten grain-production safety.In paddy fields, agriculture
The people generally prevent and treat Population of Rice Brown Planthopper, chemical prevention with various insecticides (organic phosphates, carbamate, anabasine compound)
And always as the most effective approach of preventing and treating brown paddy plant hopper.For a long time, it is that the various insecticides of the various rice grubs of preventing and treating replace
Use, causing target pest to develop immunity to drugs with non-target insects stimulates reproduction.Brown paddy plant hopper is a kind of typical insecticide induction
Type rampant type insect again.Therefore, it is current very asking in the urgent need to solution to prevent and treat brown paddy plant hopper rampant again new method and new way
Topic.
RNAi technology is that the non-transcribed fragment of the 3 ' ends of the mRNA or mRNA by target gene of degrading is combined and prevents base
The expression of cause makes specific gene that a kind of highly conserved mechanisms of gene regulation of silence to occur.As RNAi has target special
Property, high efficiency, persistence, agility the advantages of, be widely used in it is various biology gene functions research in.It is anti-in insect
Field is controlled, RNAi technology is made a kind of next new strategy of insect pest control and gradually received by people in recent years, and in laboratory bar
The insects such as Lepidoptera, coleoptera, Semiptera are successfully applied under part.In RNAi researchs, suitable target gene and appropriate lead
It is its successful key to enter method.In entomology research, some Lepidoptera elder brothers (are widely used in using micro-injection mainly now
Worm), feeding method (the less piercing-sucking mouthparts pests of build) and genetically modified plants method.For these reasons, as long as finding properly
PHF target genes, then using RNAi technology just can be good at control pest population growth.Both will not using RNAi technology
Make insect develop immunity to drugs, will not also stimulate pest resurgence, the operation and maintenance ecological balance of agricultural chemicals can be reduced again, improve agriculture
Crop product safety.Additionally, the control differred primarily in that from the research of male sterility angle to pest population length of our researchs
Effect is evaluated, and can control determining for Population of Rice Brown Planthopper growth as evaluating by these important reproductive parameters indexs
Figureofmerit.
The content of the invention
It is an object of the invention to provide in brown paddy plant hopper room RNAi artificial diet techniques and quantitative assessment control effect method.
For reaching above-mentioned purpose, the technical scheme that the present invention is provided is：
A kind of raising and evaluation method based on male sterility after RNAi to Population of Rice Brown Planthopper control effect, using RNAi skills
Art control male worm infertility, the male worm and female adult post-coitum for then recycling infertility detect the reproductive parameters of its male worm or female adult to determine
The control effect that amount assessment Population of Rice Brown Planthopper increases, method for breeding comprise the steps：
(1) dsRNA design of primers：According to known NlPHF7 genes (NLU00303) primers, using primer
The specific thing of design software design；Every primer is evaluated and is changed, with avoid in primer or between primer dimer with
And the appearance of primer mismatch site etc., final design goes out dsPHF7 double-chain primers；
(2) extraction of brown paddy plant hopper male worm RNA and cDNA synthesis：Brown paddy plant hopper male worm total serum IgE, Ran Houhe are extracted using kit
Into cDNA；
(3) dsPHF7 synthesis：After using kit synthesis dsPHF7, purifying is quantitative determined by ultraviolet absorption spectroscopy
DsPHF7 concentration, and detected to guarantee its integrality with agarose gel electrophoresis；
(4) in Population of Rice Brown Planthopper room RNAi raising：Brown paddy plant hopper is raised by 3 age nymph with man-made feeds, double with glass
Siphunculus is sealed with Parafilm films as receptacle, both-side opening, plus 40 μ L contain the liquid of the dsPHF7 that concentration is 0.05 μ g/mL
State man-made feeds, then feed is sealed with Parafilm films.
In method for breeding step (4), each glass two-way pipe discharges 30 brown paddy plant hopper nymphs；Changed once fresh per two days
Man-made feeds.
In method for breeding step (4), glass two-way pipe is positioned over bio-incubator, is wrapped up with black cotton, using insect
Phototaxis make which arrive glass two-way pipe two ends, take food both sides and seal intermembranous feed.
Described raising and evaluation method based on male sterility after RNAi to Population of Rice Brown Planthopper control effect：Evaluation method
Comprise the steps：
(1) brown paddy plant hopper 3 age nymph RNAi was raised to 5 latter stages in age, is suctioned out to be positioned over and is placed with same tillering stage paddy rice
In the glass of cane, raised to adult is just sprouted wings into, the firm adult for sprouting wings carried out into a female hero then and is matched,
The paddy rice rice stem for placing fresh tillering stage carries out oviposition；
(2) a paddy rice stem was changed every 2 days, is then dissected under the microscope and is calculated its each egg laying amount, whole
Process record female adult preoviposition period, egg-laying season, life-span and the index of aging of lay eggs sum and male worm；Each treatment group index and right
Repeat according at least 20 biology of group index；
(3) measure of reproductive parameters index was carried out to the male worm of 1 day, 3 days and 5 days after emergence, to post-coitum 2 days and 3 days
Female adult carry out the measure of reproductive parameters index, then dissect female, male worm internal genitalia system, whether observe in which reproductive system
Deformity；
(4) egg laying amount of the reproductive parameters and female adult of the female male worm for the treatment of group accurately can be evaluated compared with control group
Control effect of the male sterility to control to Population of Rice Brown Planthopper.
Wherein, after emergence, the row reproductive parameters index of the male worm of 1 day, 3 days and 5 days includes that arginine and accessory gland albumen contain
Amount；The reproductive parameters index of the post-coitum female adult of 2 days and 3 days includes egg laying amount, fat-body and ovary internal protein and Nlvg tables
Up to level.
Beneficial effect：
The invention provides RNAi artificial diet techniques and Population of Rice Brown Planthopper quantitative assessment control are imitated in a kind of brown paddy plant hopper room
The method of fruit.In terms of RNAi artificial feedings in brown paddy plant hopper room, main purpose is using RNAi technology silence male worm reproductive gene
NlPHF7, designs specificity dsRNA double-chain primers (primer is as shown in table 1) according to NlPHF7 genes (NLU00303), recycles
The dsRNA double-strands (Fig. 1) of T7 RiboMax RNAi System (Progema, Madism, WI, USA) synthesizing and purifyings, purifying
During dsPHF7 double-strands (final concentration of 0.05 μ g/mL) add man-made feeds afterwards, then brown paddy plant hopper 3 age nymph is carried out in RNAi rooms
Raise.
The method of Population of Rice Brown Planthopper quantitative assessment its control effect, essentially consists in RNAi technology silence brown paddy plant hopper male worm target
After gene, after the firm emerging adult of brown paddy plant hopper, female, male imago mates respectively or individually raises, and then determines female, male worm reproduction ginseng
Number, by treatment group and the accessory gland protein content and arginine content, the egg laying amount of female adult and Nlvg genes of control group male worm
The reproductive parameters such as expression multiple compare, and can be good at evaluating control effect of the male sterility to brown paddy plant hopper, and these are important
The measurement index whether reproductive parameters can increase as Population of Rice Brown Planthopper, the preventing and treating for brown paddy plant hopper provide new way and Xin Fang
Method.
Description of the drawings
Fig. 1：Invention is using NlPHF7 genes design specific primer synthesis dsPHF7 electrophoresis detection collection of illustrative plates.(in figure：1. just
Adopted ssRNA；2. antisense ssRNA；3. double-strand dsPHF7).
Fig. 2：Raised using RNAi in the brown paddy plant hopper dsPHF7 rooms of synthesizing and purifying of the present invention and female male worm mating flow chart.
(in figure：A. brown paddy plant hopper RNAi indoor feedings and Population Control effect diagram；B. brown paddy plant hopper RNAi indoor feedings；C. brown paddy plant hopper
Take food dsRAN+ man-made feeds；D. the female male worm mating just sprouted wings；E. the female mating of Dan Xiongdan).
Fig. 3：Using male worm and the female male worm internal genitalia comparison diagram of female adult post-coitum of this RNAi feeding.(in figure：A.
Female adult ovary (is compareed)；The female adult ovary (negative control) of B.dsGFP process；The female adult ovary of C.dsNlPHF7 process；D. it is male
Worm internal genitalia (is compareed)；E.dsGFP processes male worm internal genitalia (negative control)；Reproduction in the male worm of F.dsNlPHF7 process
Device).
Specific embodiment
With reference to specific embodiment, the invention will be further described.
1.dsRNA design of primers
According to known NlPHF7 genes (NlU00303) primers.According to design of primers principle and attention thing
, carry out the design of primer as far as possible in specific regions, using Primer primer 5.0, Oligo 6.71 and Primer
The specific thing of 3.0online primer-design softwares design, then every primer is commented with 6.71 primer-design softwares of Oligo
Valency and modification, to avoid appearance of dimer and primer mismatch site etc. in primer or between primer, final design goes out
DsPHF7 double-chain primers (table 1)
2. the extraction of brown paddy plant hopper male worm RNA and cDNA synthesis
This experiment using Progega companies of U.S. SV total isolation system Kit (Promega,
Corporation, Madison, WI, USA) kit extraction brown paddy plant hopper male worm total serum IgE.The method is simple to operate, quick, efficient
And it is nontoxic, the RNA of extraction meets this research requirement, comprises the following steps that：
1) after sprout wings 5, the brown paddy plant hopper male worm of 2 days is placed in 1.5mL centrifuge tubes, add 175 μ L RNA Lysis to from
Grinding homogenate in heart pipe, adds RNA Dilution Buffer (blueness) of 350 μ L, overturns 3-4 mixing.70 DEG C of water-baths
3min；
2) 12,000 × g are centrifuged 10min, and supernatant is moved in new 1.5mL centrifuge tubes；
3) 95% ethanol, 200 μ L are added, is mixed 3-4 time with liquid-transfering gun, be transferred in centrifugal column assembly, 12,000 × g
Centrifugation 1min；
4) liquid in collecting pipe is discarded, collecting pipe is reclaimed, 600 μ L RNA Wash Solution, 12,000 × g are added
Centrifugation 1min；
5) liquid in collecting pipe is discarded, and 40 μ L Yellow Core Buffer and 5 μ L is added to centrifugal column inner membrance
0.09M Mncl2And 5 μ L DNase I mixtures incubated (now with)；
6) 15min is incubated under room temperature (25 DEG C) environment, be subsequently adding 200 μ L DNAase Stop Solution, 12,000
× g is centrifuged 1min；
7) 600 μ L RNA Wash Solution are directly added into and then, 12,000 × g centrifugation 1min；
8) liquid is discarded, adds 250 μ L RNA Wash Solution, 12,000 × g centrifugation 2min into centrifugal column；
9) centrifugal column is moved to and eluted on wash-out pipe, 40 μ L Nuclease-free are added to post central authorities
Water, is overlying on film surface, and after standing 1min, 12,000 × g centrifugation 1min, using -80 DEG C of preservations of front placement.
According to the first chain cDNA kit PrimeScriptTM 1st stand cDNA synthesis Kit(TaKaRa
Biotechnology (Dalian) Co., Ltd) synthesis cDNA.
3.dsPHF7 synthesizes
The synthesis of double-strand dsPHF7 is according to T7 RiboMAXTM Express RNAi System(Progema
Corporation, Madison, WI, USA) kit.Comprise the following steps that：
1) (single-stranded) synthesis of justice ssRNA, reaction system are as follows：
2) (single-stranded) synthesis of antisense ssRNA, reaction system are as follows
PCR reaction conditions are：94 DEG C of denaturations 5min, 94 DEG C of denaturation 40S, 55 DEG C (Tm) annealing 40S, 72 DEG C of extensions
1min, totally 30 circulations, last 72 DEG C of extensions 10min.
DsRNA is further synthesized according to the ssRNA of PCR reaction conditions synthesis
37 DEG C of water-bath 4h after mentioned reagent mixing.Then respectively will be the positive-sense strand ssRNA and antisense ssRNA of above-mentioned synthesis each
Annealing reaction is carried out after 20 μ L mixing, and 70 DEG C of incubation 10min are subsequently cooled to room temperature about 20min, and total reaction volume is 40 μ L.
2 μ L didution RNAase A solution (1 are placed again:200) it is anti-that He 2 μ L RNase-free DNase are added to transcription
In answering the centrifuge tube of mixed liquor, 37 DEG C of water-bath 30min, then 110 μ L, 95% ethanol and 4.4 μ L 3M sodium acetates (pH5.2) are placed,
12,000 × g is centrifuged 1min, then washs precipitation with 70% ethanol of 0.5mL, then drying at room temperature, with 50 μ L Nuclear-
Free water dissolve.DsPHF7 (260nm) concentration of ultraviolet absorption spectroscopy quantitative determination purifying, and it is electric with Ago-Gel
Swimming detects to guarantee its integrality.
4. in Population of Rice Brown Planthopper room RNAi raising
Brown paddy plant hopper is raised by 3 age nymph with man-made feeds, with glass two-way pipe (15 × 2.5cm) as receptacle, two
Side opening Parafilm films are sealed plus the liquid artificial containing dsPHF7 (design dsRNA, ultimate density is 0.05ng/mL) is raised
Material (40 μ L), then feed is sealed with one layer of Parafilm film.Each glass two-way pipe discharges 30 brown paddy plant hopper nymphs.Glass tube
It is positioned over bio-incubator (Ningbo of Zhejiang south of the River Instrument Ltd.) (temperature：26±2℃；Humidity is 75-80%；Illumination is
16L:8D), (Fig. 2) is wrapped up with black cotton, make which arrive glass tube two ends using the phototaxis of insect, take food both sides sealing intermembranous
Feed.Fresh man-made feeds were changed once per two days.
5. Population of Rice Brown Planthopper increases the method for quantitatively evaluating of control effect
Brown paddy plant hopper 3 age nymph RNAi was raised to 5 latter stages in age (about 8～10 days), suctioned out be positioned over be placed with it is same
Raised to adult is just sprouted wings in the glass (high 12cm, bore 10cm) of tillering stage paddy rice cane, then just will be sprouted wings
Adult carry out a female hero and carry out matching placing the paddy rice rice stem in fresh tillering stage and carrying out oviposition.One was changed every 2 days
Secondary paddy rice stem, then under the microscope dissect calculate its each egg laying amount, whole process can record female adult preoviposition period,
The indexs such as the life-span in egg-laying season, life-span and spawning sum and male worm.Each processes and compares at least 20 biology and repeats.To plumage
After change, the male worm of 1 day, 3 days and 5 days carries out the measure of reproductive parameters index (arginine and accessory gland protein content), post-coitum 2 days
The measure of reproductive parameters index (egg laying amount, fat-body and ovary internal protein and Nlvg expressions) is carried out with the female adult of 3 days,
Then female, male worm internal genitalia system is dissected, reproductive system in which is observed whether lopsided (Fig. 3).The life of the female male worm for the treatment of group
Grow the egg laying amount of parameter and female adult compared with the control, can accurately evaluate control of the male sterility to control to Population of Rice Brown Planthopper
Effect.
The above, is only presently preferred embodiments of the present invention, not makees any pro forma restriction to the present invention, any ripe
Professional and technical personnel is known, it is in the range of without departing from technical solution of the present invention, according to the technical spirit of the present invention, real to more than
Apply any simple modification, equivalent and improvement that example is made etc., still fall within technical solution of the present invention protection domain it
It is interior.
Claims (5)
1. a kind of raising and evaluation method based on male sterility after RNAi to Population of Rice Brown Planthopper control effect, it is characterised in that：
Using RNAi technology control male worm infertility, the male worm and female adult post-coitum that then recycle infertility detect the life of its male worm or female adult
The control effect that parameter carrys out the growth of qualitative assessment Population of Rice Brown Planthopper is grown, method for breeding comprises the steps：
(1) dsRNA design of primers：According to known NlPHF7 genes (NLU00303) primers, using design of primers
Software for Design specificity thing；Every primer is evaluated and changed, to avoid in primer or between primer dimer and draw
Appearance of thing mismatch site etc., final design go out dsPHF7 double-chain primers；
(2) extraction of brown paddy plant hopper male worm RNA and cDNA synthesis：Brown paddy plant hopper male worm total serum IgE is extracted using kit, is then synthesized
cDNA；
(3) dsPHF7 synthesis：After using kit synthesis dsPHF7, purifying is quantitative determined by ultraviolet absorption spectroscopy
DsPHF7 concentration, and detected to guarantee its integrality with agarose gel electrophoresis；
(4) in Population of Rice Brown Planthopper room RNAi raising：Brown paddy plant hopper is raised by 3 age nymph with man-made feeds, uses glass two-way pipe
Used as receptacle, both-side opening is sealed with Parafilm films, plus 40 μ L contain the liquid people of the dsPHF7 that concentration is 0.05 μ g/mL
Work feed, then feed is sealed with Parafilm films.
2. raising and evaluation side based on male sterility after RNAi to Population of Rice Brown Planthopper control effect as claimed in claim 1
Method, it is characterised in that：In method for breeding step (4), each glass two-way pipe discharges 30 brown paddy plant hopper nymphs；Changed once per two days
Fresh man-made feeds.
3. raising and evaluation side based on male sterility after RNAi to Population of Rice Brown Planthopper control effect as claimed in claim 1
Method：In method for breeding step (4), glass two-way pipe is positioned over bio-incubator, is wrapped up with black cotton, using the light that becomes of insect
Property make which arrive glass two-way pipe two ends, take food both sides and seal intermembranous feed.
4. raising and evaluation side based on male sterility after RNAi to Population of Rice Brown Planthopper control effect as claimed in claim 1
Method：Evaluation method comprises the steps：
(1) brown paddy plant hopper 3 age nymph RNAi was raised to 5 latter stages in age, is suctioned out to be positioned over and is placed with same tillering stage paddy rice cane
Glass in, raised to adult is just sprouted wings into, the firm adult for sprouting wings carried out into a female hero then and is matched, placed
The paddy rice rice stem in fresh tillering stage carries out oviposition；
(2) a paddy rice stem was changed every 2 days, is then dissected under the microscope and is calculated its each egg laying amount, in whole process
Record female adult preoviposition period, egg-laying season, life-span and the index of aging of lay eggs sum and male worm；Each index at least carries out 20 lifes
Thing repeats；
(3) measure of reproductive parameters index is carried out to the male worm of 1 day, 3 days and 5 days after emergence, to the female of post-coitum 2 days and 3 days
Worm carries out the measure of reproductive parameters index, then dissects female, male worm internal genitalia system, whether abnormal observes reproductive system in which
Shape；
(4) egg laying amount of the reproductive parameters and female adult of the female male worm for the treatment of group can accurately evaluate male compared with control group
Control effect of the infertility to control to Population of Rice Brown Planthopper.
5. raising and evaluation side based on male sterility after RNAi to Population of Rice Brown Planthopper control effect as claimed in claim 4
Method, it is characterised in that：After emergence, the row reproductive parameters index of the male worm of 1 day, 3 days and 5 days includes that arginine and accessory gland albumen contain
Amount；The reproductive parameters index of the post-coitum female adult of 2 days and 3 days includes egg laying amount, fat-body and ovary internal protein and Nlvg tables
Up to level.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106942162A (en) * | 2017-04-25 | 2017-07-14 | 遵义医学院 | A kind of device and its application method suitable for white backed planthopper feeding method RNA interference experiments |
| CN107494451A (en) * | 2017-09-09 | 2017-12-22 | 合肥天敌之家生物技术有限公司 | A kind of acquisition methods of small new neoseiulus californicus reproductive system |
| CN108260566A (en) * | 2018-01-17 | 2018-07-10 | 西南大学 | A kind of RNA interference methods of l.bostrychophila SOD1 genes |
| CN110117619A (en) * | 2018-02-05 | 2019-08-13 | 中国科学院上海生命科学研究院 | A kind of method preparing diamondback moth male-sterile line and its nucleic acid |
| CN114540409A (en) * | 2022-02-14 | 2022-05-27 | 浙江大学 | Application of brown planthopper gene fragment NlVgN in improving insect resistance of rice |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102220339A (en) * | 2011-05-12 | 2011-10-19 | 南京农业大学 | Laodelphax striatellus lethal gene fragment Ribosomal protein L9-B based on gene-silencing technology and dsRNA thereof |
-
2016
- 2016-10-17 CN CN201610905214.2A patent/CN106520768B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102220339A (en) * | 2011-05-12 | 2011-10-19 | 南京农业大学 | Laodelphax striatellus lethal gene fragment Ribosomal protein L9-B based on gene-silencing technology and dsRNA thereof |
Non-Patent Citations (3)
| Title |
|---|
| LIN-QUAN GE ET AL.: "Proteomic Analysis of Insecticide Triazophos-Induced Mating-Responsive Proteins of Nilaparvata lugens Stål (Hemiptera: Delphacidae)", 《J. PROTEOME RES.》 * |
| LIN-QUAN GE ET AL.: "Silencing a sugar transporter gene reduces growth and fecundity in the brown planthopper，Nilaparvata lugens (Stål) (Hemiptera: Delphacidae)", 《SCIENTIFIC REPORTS》 * |
| 李鑫: "三唑磷处理后丙酮酸激酶和己糖激酶基因沉默对褐飞虱生殖的影响", 《中国优秀硕士学位论文全文数据库农业科技辑》 * |
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| CN110117619A (en) * | 2018-02-05 | 2019-08-13 | 中国科学院上海生命科学研究院 | A kind of method preparing diamondback moth male-sterile line and its nucleic acid |
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