CN111004852A - Method for screening resistance of bombyx mori nuclear polyhedrosis virus by using BmIML-2 gene - Google Patents
Method for screening resistance of bombyx mori nuclear polyhedrosis virus by using BmIML-2 gene Download PDFInfo
- Publication number
- CN111004852A CN111004852A CN202010005619.7A CN202010005619A CN111004852A CN 111004852 A CN111004852 A CN 111004852A CN 202010005619 A CN202010005619 A CN 202010005619A CN 111004852 A CN111004852 A CN 111004852A
- Authority
- CN
- China
- Prior art keywords
- silkworm
- nuclear polyhedrosis
- polyhedrosis virus
- bmiml
- gene
- Prior art date
- 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.)
- Granted
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/124—Animal traits, i.e. production traits, including athletic performance or the like
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/166—Oligonucleotides used as internal standards, controls or normalisation probes
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Analytical Chemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Immunology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a method for screening resistance of silkworm nuclear polyhedrosis viruses by using BmIML-2 genes, which is characterized in that the nuclear polyhedrosis viruses are added when mulberry is fed for the last time before mounting, and the transcription level of the BmIML-2 genes is used as a detection index to judge the strength of the resistance of silkworms to the viruses. Compared with the prior art, the invention has the following advantages: (1) the method selects 5-age cocoon mounting and pre-poison adding, can shorten the test period, has small workload and reduces the probability of mutual cross infection among silkworms; (2) the resistance of the silkworm variety screened by the method of the invention to the nuclear polyhedrosis virus is improved by 10 to 100 times; (3) the silkworm variety screened by the method provided by the invention is stable in resistance and strong in physique after being bred for multiple generations.
Description
Technical Field
The invention belongs to the technical field of improved silkworm variety breeding, relates to a technology for breeding antiviral silkworm varieties by adopting a genetic engineering method, and particularly relates to a method for screening resistance of silkworm nuclear polyhedrosis virus by utilizing BmIML-2 gene.
Background
The silkworm as an important economic insect and a model insect has great significance in scientific research, silk production, promotion of farmer income increase and the like. The silkworm nuclear polyhedrosis virus-induced silkworm hemopus diseases are silkworm diseases with the most serious harm in the silkworm industry production, the disease has extremely strong infectivity and is difficult to prevent and control, and the economic loss caused by the disease accounts for 70-80% of the total loss of the silkworm diseases. The breeding of resistant varieties of silkworm nuclear polyhedrosis virus is always an important research direction in the industry, and the screening of key genes of the resistance of silkworm nuclear polyhedrosis virus is also one of important works of researchers for breeding new resistant varieties of silkworm. The traditional screening method is used for feeding the viruses in larva stages such as 2-instar or 3-instar of silkworms, and the method is easy to pollute the silkworm breeding environment in the silkworm stage to cause mutual infection among the silkworms.
Disclosure of Invention
The technical problem to be solved is as follows: in order to overcome the defects of the prior art, the invention feeds the silkworm nuclear polyhedrosis virus when the mulberry is fed for the last time before mounting, and selects the variety with high resistance to the nuclear polyhedrosis virus by taking the transcription level of immune related genes as a detection index; in view of the above, the present invention provides a method for screening resistance to nuclear polyhedrosis virus of silkworms using the BmIML-2 gene.
The technical scheme is as follows: a method for screening resistance to bombyx mori nuclear polyhedrosis virus using BmIML-2 gene, the method comprising the steps of:
(1) selecting at least one silkworm variety as a test material and feeding the silkworm variety to the 5 th instar; soaking fresh mulberry leaves in a nuclear polyhedrosis virus solution;
(2) when the mulberry is fed for the last time before cocooning, the mulberry leaves in the step (1) are fed to the silkworm larvae of each variety, then the silkworm larvae are continuously fed until cocooning and cocooning are carried out, pupae which die from diseases are eliminated in the period, and seeds are produced after moth is sent out;
(3) taking the silkworm moth produced in the step (2), extracting RNA in a tissue and carrying out reverse transcription to obtain cDNA, designing a primer according to the BmIML-2 gene full-length sequence, and detecting the transcription level difference of the BmIML-2 gene of the silkworm in each silkworm variety by RT-PCR (reverse transcription-polymerase chain reaction) by taking the cDNA as a template;
(4) screening varieties with remarkably up-regulated BmIML-2 gene transcription level after the nuclear polyhedrosis virus is added and silkworm eggs produced by individuals for stock subculture according to the RT-PCR detection result in the step (3);
(5) and (5) repeating the steps (1) to (4), and repeatedly screening the subcultured silkworm variety for more than 6 generations to obtain the silkworm nuclear polyhedrosis virus disease resistant variety.
Preferably, the silkworm varieties selected in the step (1) are irradiant, white jade C3VR6, white jade CB, PAB and pinus sylvestris.
Preferably, the concentration of the nuclear polyhedrosis virus solution in step (1) is 1X 106-5×107one/mL.
Preferably, the primer sequence in step (3) is:
F(SEQ ID NO.1)：5’-AGAGTTCGTGTGGCATCTA-3’，
R(SEQ ID NO.2)：5’-ACCATATGAGAGGCAGGGTT-3’。
preferably, the tissue in step (3) is adipose.
Preferably, the reference gene detected by RT-PCR in step (3) is BmGAPDH gene.
Preferably, the resistance of the bombyx mori variety nuclear polyhedrosis virus obtained in the step (5) is verified: hatching the silkworm eggs which are reserved for subculture, feeding the silkworm eggs to the 2 th instar of the silkworm, then adding the nuclear polyhedrosis virus, and continuously feeding the silkworm eggs to the 4 th instar of the silkworm, and counting the survival rate.
Preferably, the following silkworm varieties: the resistance of the wave illumination, the white jade C3VR6, the white jade CB and the PAB to the nuclear polyhedrosis virus is 10 to 100 times higher than that of the pinus sylvestris.
The principle/thought of the design of the method provided by the invention is as follows:
the C-type lectin plays a key role as an important pattern recognition receptor in the process of starting the natural immune response of insects, killing and eliminating exogenous pathogens. The BmIML-2 serving as a C-type lectin gene has the highest transcription level in silkworm fat bodies, and the transcription level is obviously up-regulated after the addition of the polyhedrosis virus, so that the BmIML-2 plays an important role in the process of identifying and resisting exogenous virus infection of the silkworms. Therefore, the invention uses the mutant as a detection index of resistance of the silkworm nuclear polyhedrosis virus to screen the nuclear polyhedrosis virus high-resistance variety.
The method disclosed by the invention is characterized in that BmNPV is added when the mulberry is fed for the last time before mounting, and the resistance of the silkworm to the virus is judged by taking the transcription level of the BmIML-2 gene as a detection index. In addition, the virus adding before cocooning has the advantages of short experimental period, small workload and easy operation, and the silkworm can not defecate any more after pupating, thereby effectively avoiding cross contamination caused by adding the virus too early; in addition, sampling detection is carried out after the adult moths are bred, the silkworm moths are mated and spawned at the moment, the seed production is completed, and silkworm seeds are reserved for continuously breeding nuclear polyhedrosis virus resistant varieties. The transcription level difference of the BmIML-2 gene in various varieties and individuals of the silkworm is detected by RT-PCR technology, and the method is easy to operate, convenient and stable. The BmIML-2 gene transcription level is used as a detection index, and the method has a good indication effect on the strength of the resistance of the silkworm variety fed with the polyhedrosis virus, so that the variety with strong resistance can be effectively selected.
Has the advantages that: (1) the method selects 5-age cocoon mounting and pre-poison adding, can shorten the test period, has small workload and reduces the probability of mutual cross infection among silkworms; (2) the resistance of the silkworm variety screened by the method of the invention to BmNPV is improved by 10-100 times; (3) the silkworm variety screened by the method provided by the invention is stable in resistance and strong in physique after being bred for multiple generations.
Drawings
FIG. 1 is a graph of RT-PCR detection of differences in the transcription levels of BmIML-2 gene in different tissues of Bombyx mori, wherein 1: head, 2: middle intestine, 3: fat body, 4: silk gland, 5: mahalanobis tube, 6: epidermis, 7: an air tube;
FIG. 2 is a graph showing the difference in transcription level between the BmIML-2 gene of Bombyx mori before and after feeding BmNPV in several varieties of Bombyx mori by RT-PCR, wherein 1: illumination, 2: white jade C3VR6, 3: white jade CB, 4: PAB, 5: and (4) pinus cyanine.
Detailed Description
The following examples further illustrate the present invention but are not to be construed as limiting the invention. Modifications and substitutions to methods, procedures, or conditions of the invention may be made without departing from the spirit and substance of the invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art.
Example 1 detection of the difference in transcription level of BmIML-2 Gene in various tissues of Larvae by RT-PCR
Extracting RNA of head, midgut, fat body, silk gland, Marek's tube, epidermis and trachea of 5-year-old 3-day silkworm larva, and reverse transcribing to obtain cDNA. Designing a specific primer by taking cDNA obtained by reverse transcription as a template, wherein the forward primer (SEQ ID NO. 1): 5'-AGAGTTCGTGTGGCATCTA-3', reverse primer (SEQ ID NO.2): 5'-ACCATATGAGAGGCAGGGTT-3'. Silkworm BmGAPDH gene is used as an internal reference gene, and the upstream and downstream primer sequences are F (SEQID NO. 3): 5'-TTCATGCCACAACTGCTACA-3', R (SEQ ID NO. 4): 5'-AGTCAGCTTGCCATTAAGAG-3', detecting the difference of the transcription level of BmIML-2 gene in different tissues of silkworm larva (shown in figure 1).
Example 2 several silkworm varieties were treated with nuclear polyhedrosis virus and examined for changes in BmIML-2 transcript levels
When the mulberry is fed for the first time before silkworm larvae are cocooned, a plurality of silkworm varieties are used as experimental materials, and all the varieties are divided into two groups for treatment. The treatment group was supplemented with nuclear polyhedrosis virus (concentration 1X 10)7one/mL) of the soaked fresh mulberry leaves, and the control group was fed with fresh mulberry leaves without soaking in virus. And after adding poison, immediately eliminating the dead silkworms until cocooning and cocooning are carried out, eliminating the dead silkworms, and carrying out female and male number-matching mating and seed production. Taking each group of silkworm moths after the production of the seeds of the silkworms, extracting fat body RNA and carrying out reverse transcription to obtain cDNA. Specific primers (same as example 1) are designed, cDNA obtained by reverse transcription is used as a template, and the difference of transcription levels of the bombyx mori BmIML-2 gene before and after virus addition in the bombyx mori variety is detected. The detection results are shown in figure 2, wave illumination, white jade C3VR6, and white jadeAfter the CB and PAB varieties are fed with viruses, the BmIML-2 transcription level is obviously increased; the BmIML-2 transcription level is not obviously different before and after the pinus sylvestris variety is fed with the virus.
Example 3 continuous screening of silkworm varieties having significantly upregulated BmIML-2 transcript levels after viral feeding
According to the detection result in the embodiment 2, silkworm eggs produced by silkworm variety individuals with significantly-increased BmIML-2 transcription level after feeding of nuclear polyhedrosis virus are selected for seed reservation and subculture, and the subcultured silkworm varieties are repeatedly screened. Namely, when the mulberry is fed for the first time before silkworm larvae are clustered, the feeding virus treatment in the previous step is repeated, the difference of the transcription levels of the BmIML-2 genes before and after feeding the virus is detected by using an RT-PCR method, and excellent male and female individuals are screened for seed production and subculture propagation. Through the screening by the method, the resistance of the variety of the Baiyu CB to the nuclear polyhedrosis virus is improved by 10-100 times. Feeding nuclear polyhedrosis virus (1X 10) to Jade CB and Cyanina pine at the beginning of 2-year-old silkworm7One per mL) and examined in 4-instar silkworms. The results show that more than 90% of white jade CB survives healthily, and the pinus sylvestris almost all suffers from diseases and dies. The bred Baiyu CB variety has stable resistance and strong physique and obviously improves the resistance to the nuclear polyhedrosis virus. The method for screening the resistance of the silkworm nuclear polyhedrosis virus disease is feasible and has obvious effect, and can be applied to the breeding production practice of the resistance of the silkworm nuclear polyhedrosis virus disease.
The difference of the transcription level of the gene in resistant and non-resistant varieties of the bombyx mori nuclear polyhedrosis virus is detected by taking BmIML-2 as a detection index. After the nuclear polyhedrosis virus is added, the transcription level of the BmIML-2 gene in a resistant variety is obviously increased, but the BmIML-2 gene is not obviously changed in a non-resistant variety, which shows that the BmIML-2 gene plays a key role in the process of resisting the invasion of the polyhedrosis virus by silkworms and can be used as an indicator gene for the breeding of the nuclear polyhedrosis virus resistance of the silkworms in the subsequent breeding practice.
Sequence listing
<110> university of Jiangsu science and technology
<120> method for screening resistance of silkworm nuclear polyhedrosis virus using BmIML-2 gene
<160>4
<170>SIPOSequenceListing 1.0
<210>1
<211>19
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>1
agagttcgtg tggcatcta 19
<210>2
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>2
accatatgag aggcagggtt 20
<210>3
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>3
ttcatgccac aactgctaca 20
<210>4
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>4
agtcagcttg ccattaagag 20
Claims (8)
1. A method for screening resistance of silkworm nuclear polyhedrosis virus by using BmIML-2 gene, which is characterized by comprising the following steps:
(1) selecting at least one silkworm variety as a test material and feeding the silkworm variety to the 5 th instar; soaking fresh mulberry leaves in a bombyx mori nuclear polyhedrosis virus BmNPV solution;
(2) when the mulberry is fed for the last time before cocooning, the mulberry leaves in the step (1) are fed to the silkworm larvae of each variety, then the silkworm larvae are continuously fed until cocooning and cocooning are carried out, pupae which die from diseases are eliminated in the period, and seeds are produced after moth is sent out;
(3) taking the silkworm moth produced in the step (2), extracting RNA in a tissue and carrying out reverse transcription to obtain cDNA, designing a primer according to the BmIML-2 gene full-length sequence, and detecting the transcription level difference of the BmIML-2 gene of the silkworm in each silkworm variety by RT-PCR (reverse transcription-polymerase chain reaction) by taking the cDNA as a template;
(4) screening varieties with the BmIML-2 gene transcription level remarkably increased after BmNPV is fed and silkworm eggs laid by individuals for subculture according to the RT-PCR detection result in the step (3);
(5) and (5) repeating the steps (1) to (4), and repeatedly screening the subcultured silkworm variety for more than 6 generations to obtain the silkworm nuclear polyhedrosis virus disease resistant variety.
2. The method for screening resistance to nuclear polyhedrosis virus of silkworms according to claim 1, wherein the silkworm variety selected in step (1) is selected from the group consisting of radiogram, albedo C3VR6, albedo CB, PAB and pinus sylvestris.
3. The method for screening resistance to nuclear polyhedrosis virus of silkworms according to claim 1, wherein the concentration of the BmNPV solution in step (1) is 1X 106-5×107one/mL.
4. The method for screening resistance to nuclear polyhedrosis virus of silkworms according to claim 1 using BmIML-2 gene, wherein the primer sequence in step (3) is:
F(SEQ ID NO.1)：5’-AGAGTTCGTGTGGCATCTA-3’，
R(SEQ ID NO.2)：5’-ACCATATGAGAGGCAGGGTT-3’。
5. the method for screening resistance to nuclear polyhedrosis virus of silkworms according to claim 1 wherein the tissue in step (3) is an adipose tissue.
6. The method for screening resistance to nuclear polyhedrosis virus of silkworms according to claim 1, wherein the BmGAPDH gene is used as an internal reference gene for RT-PCR detection in step (3).
7. The method for screening resistance to nuclear polyhedrosis virus of silkworms according to claim 1 using BmIML-2 gene, wherein the resistance to nuclear polyhedrosis virus of silkworm variety obtained in step (5) is verified: hatching the silkworm eggs reserved for the subculture, feeding the silkworm eggs to the 2 th instar of the silkworm, then adding BmNPV, and continuously feeding the silkworm eggs to the 4 th instar of the silkworm, and then counting the survival rate.
8. The method for screening resistance to nuclear polyhedrosis virus of silkworms according to claim 2 or 7 using BmIML-2 gene, characterized in that the following silkworm varieties: the resistance of the wave illumination, the white jade C3VR6, the white jade CB and the PAB to the nuclear polyhedrosis virus is 10 to 100 times higher than that of the pinus sylvestris.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010005619.7A CN111004852B (en) | 2020-01-03 | 2020-01-03 | Method for screening silkworm nuclear polyhedrosis virus disease resistance by using BmIML-2 gene |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010005619.7A CN111004852B (en) | 2020-01-03 | 2020-01-03 | Method for screening silkworm nuclear polyhedrosis virus disease resistance by using BmIML-2 gene |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN111004852A true CN111004852A (en) | 2020-04-14 |
| CN111004852B CN111004852B (en) | 2023-07-25 |
Family
ID=70120412
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010005619.7A Active CN111004852B (en) | 2020-01-03 | 2020-01-03 | Method for screening silkworm nuclear polyhedrosis virus disease resistance by using BmIML-2 gene |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN111004852B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005333913A (en) * | 2004-05-28 | 2005-12-08 | National Institute Of Agrobiological Sciences | Transgenic silkworm exhibiting resistance against nucleopolyhedrovirus and method for producing the same |
| CN101558754A (en) * | 2009-05-31 | 2009-10-21 | 安徽农业大学 | Method for select-breeding anti-virosis Bombyx mori varieties by utilizing Thailand Bombyx mori disease-resistant gene |
| CN104839099A (en) * | 2015-04-17 | 2015-08-19 | 江苏科技大学 | Method for improving silkworm nuclear polyhedrosis resistance through hybridization and backcross |
| CN105145500A (en) * | 2015-06-30 | 2015-12-16 | 江苏科技大学 | Bombyxmori Nucleopolyhedrovirus ( BmNPV) full-resistance breeding method |
| CN108308127A (en) * | 2018-05-17 | 2018-07-24 | 江苏科技大学 | A method of silkworm is improved to BmCPV virus resistances |
| CN110295196A (en) * | 2019-06-17 | 2019-10-01 | 江苏科技大学 | A kind of method and its recombinant baculovirus and application extending the silkworm infection nuclear polyhedrosis virus death time |
-
2020
- 2020-01-03 CN CN202010005619.7A patent/CN111004852B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2005333913A (en) * | 2004-05-28 | 2005-12-08 | National Institute Of Agrobiological Sciences | Transgenic silkworm exhibiting resistance against nucleopolyhedrovirus and method for producing the same |
| CN1706939A (en) * | 2004-05-28 | 2005-12-14 | 独立行政法人农业生物资源研究所 | Antinuclear polyhedron virus transgenic silkworm and generating method thereof |
| CN101558754A (en) * | 2009-05-31 | 2009-10-21 | 安徽农业大学 | Method for select-breeding anti-virosis Bombyx mori varieties by utilizing Thailand Bombyx mori disease-resistant gene |
| CN104839099A (en) * | 2015-04-17 | 2015-08-19 | 江苏科技大学 | Method for improving silkworm nuclear polyhedrosis resistance through hybridization and backcross |
| CN105145500A (en) * | 2015-06-30 | 2015-12-16 | 江苏科技大学 | Bombyxmori Nucleopolyhedrovirus ( BmNPV) full-resistance breeding method |
| CN108308127A (en) * | 2018-05-17 | 2018-07-24 | 江苏科技大学 | A method of silkworm is improved to BmCPV virus resistances |
| CN110295196A (en) * | 2019-06-17 | 2019-10-01 | 江苏科技大学 | A kind of method and its recombinant baculovirus and application extending the silkworm infection nuclear polyhedrosis virus death time |
Non-Patent Citations (2)
| Title |
|---|
| XIANG-JUN RAO ET AL.: "Identification of C-type lectin-domain proteins (CTLDPs) in silkworm Bombyx mori", 《DCI》 * |
| 季小存等: "家蚕 C-型凝集素 11 基因 BmCTL11 的克隆及表达分析", 《蚕业科学》 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN111004852B (en) | 2023-07-25 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106719114B (en) | Hybrid between Fugu obscurus and Fugu rubripes and its production process | |
| CN102342264B (en) | Method for breeding mite-resistant bee species | |
| CN102286533B (en) | Insect infection method for production of proteins | |
| CN112342199B (en) | Spodoptera frugiperda virus-resistant pesticide production strain and preparation method and application thereof | |
| Zhou et al. | Effects of thelytokous parthenogenesis-inducing Wolbachia on the fitness of Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae) in superparasitised and single-parasitised hosts | |
| CN111004852B (en) | Method for screening silkworm nuclear polyhedrosis virus disease resistance by using BmIML-2 gene | |
| CN113508713A (en) | Method for breeding trichogramma by using eggs of tuber moths of potatoes | |
| CN104904638B (en) | A cultivation method for carassius auratus gynogenesis fries | |
| CN114592075B (en) | Detection method of chimeric gonads after rice field eel germ cell xenograft and transplantation | |
| CN114568384B (en) | Seed production method of early-maturing grain-saving stress-resistant broiler chicken complete set system | |
| Hegazi et al. | The calyx fluid of Microplitis rufiventris parasitoid and growth of its host Spodoptera littoralis larvae | |
| CN109618811B (en) | Industrialized artificial cultivation method for cordyceps sinensis | |
| Cicero | Composite, haustellate mouthparts in netwinged beetle and firefly larvae (Coleoptera, Cantharoidea: Lycidae, Lampyridae) | |
| CN111500581A (en) | Molecular breeding method for thickening muscle between silver carps and bighead carps | |
| CN1234871C (en) | Construction method using detoxiase gene as stable expression system in silkworm | |
| Mazurkevych et al. | Cellular Composition of the Lymphoid Tissue of the Cecal Immune Formations in Ducks. | |
| Sharma et al. | A Comparative Study on the Rearing Performance of Six Strains of Eri Silk Worm Samia Ricini, Donovan in Four Different Seasons | |
| CN108308127A (en) | A method of silkworm is improved to BmCPV virus resistances | |
| CN112011540B (en) | Silkworm binary transgenic system for knocking down Seroin1 gene and preparation method of silkworm pure naked pupa variety | |
| CN116426534B (en) | Brown planthopper NlsNPF gene and application of dsRNA thereof in control of brown planthopper | |
| CN117106052B (en) | Application of dopamine transporter in regulation and control of individual division of solenopsis invicta workers | |
| CN111066700B (en) | Application of salidroside in combination with isoniazid in prolonging survival time of sea branch bacterial plaque horse fish model | |
| CN114317549B (en) | Application of muscarinic C-type acetylcholine receptor in prevention and treatment of migratory locust | |
| CN1216986C (en) | Method for expressing modified human alpha antitrypsin gene in sheep's mammar gland | |
| CN111820185B (en) | Propagation method of IFN-gamma gene defect homozygote mice and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |