CN113755530B - 敲除或沉默蜂巢小甲虫气味结合蛋白基因1在防治蜂巢小甲虫中的应用 - Google Patents
敲除或沉默蜂巢小甲虫气味结合蛋白基因1在防治蜂巢小甲虫中的应用 Download PDFInfo
- Publication number
- CN113755530B CN113755530B CN202111183680.1A CN202111183680A CN113755530B CN 113755530 B CN113755530 B CN 113755530B CN 202111183680 A CN202111183680 A CN 202111183680A CN 113755530 B CN113755530 B CN 113755530B
- Authority
- CN
- China
- Prior art keywords
- bee hive
- binding protein
- protein gene
- silencing
- beetles
- 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.)
- Active
Links
- 241000254173 Coleoptera Species 0.000 title claims abstract description 44
- 108010000645 odorant-binding protein Proteins 0.000 title claims abstract description 21
- 230000030279 gene silencing Effects 0.000 title claims abstract description 16
- 230000002265 prevention Effects 0.000 title claims description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 claims abstract description 19
- 102000040650 (ribonucleotides)n+m Human genes 0.000 claims abstract description 16
- 239000002773 nucleotide Substances 0.000 claims abstract description 4
- 125000003729 nucleotide group Chemical group 0.000 claims abstract description 4
- 239000003039 volatile agent Substances 0.000 claims description 11
- XIRNKXNNONJFQO-UHFFFAOYSA-N ethyl hexadecanoate Chemical compound CCCCCCCCCCCCCCCC(=O)OCC XIRNKXNNONJFQO-UHFFFAOYSA-N 0.000 claims description 6
- JYYFMIOPGOFNPK-AGRJPVHOSA-N ethyl linolenate Chemical compound CCOC(=O)CCCCCCC\C=C/C\C=C/C\C=C/CC JYYFMIOPGOFNPK-AGRJPVHOSA-N 0.000 claims description 3
- 229940090028 ethyl linolenate Drugs 0.000 claims description 3
- JYYFMIOPGOFNPK-UHFFFAOYSA-N ethyl linolenate Natural products CCOC(=O)CCCCCCCC=CCC=CCC=CCC JYYFMIOPGOFNPK-UHFFFAOYSA-N 0.000 claims description 3
- 229940067592 ethyl palmitate Drugs 0.000 claims description 3
- 101100224347 Arabidopsis thaliana DOF3.4 gene Proteins 0.000 abstract description 21
- 101100241976 Schizosaccharomyces pombe (strain 972 / ATCC 24843) obp1 gene Proteins 0.000 abstract description 21
- 241000223679 Beauveria Species 0.000 abstract description 9
- 238000012228 RNA interference-mediated gene silencing Methods 0.000 abstract description 5
- 230000009368 gene silencing by RNA Effects 0.000 abstract description 5
- 238000005516 engineering process Methods 0.000 abstract description 4
- 238000000034 method Methods 0.000 abstract description 4
- 235000013305 food Nutrition 0.000 abstract description 2
- 108090000623 proteins and genes Proteins 0.000 description 5
- 230000014509 gene expression Effects 0.000 description 4
- 238000002347 injection Methods 0.000 description 4
- 239000007924 injection Substances 0.000 description 4
- VLKZOEOYAKHREP-UHFFFAOYSA-N n-Hexane Chemical compound CCCCCC VLKZOEOYAKHREP-UHFFFAOYSA-N 0.000 description 4
- 238000012408 PCR amplification Methods 0.000 description 3
- 230000015572 biosynthetic process Effects 0.000 description 3
- 238000013461 design Methods 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 238000003786 synthesis reaction Methods 0.000 description 3
- 238000011144 upstream manufacturing Methods 0.000 description 3
- 241000218993 Begonia Species 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 241000238631 Hexapoda Species 0.000 description 2
- 241000607479 Yersinia pestis Species 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 235000012907 honey Nutrition 0.000 description 2
- 239000013612 plasmid Substances 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000012163 sequencing technique Methods 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 1
- 241000220275 Aethina Species 0.000 description 1
- 241001310985 Cordyceps nidus Species 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 108700039887 Essential Genes Proteins 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 241000257303 Hymenoptera Species 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 238000003766 bioinformatics method Methods 0.000 description 1
- 239000013599 cloning vector Substances 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 230000001418 larval effect Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 230000034267 olfactory behavior Effects 0.000 description 1
- 230000003071 parasitic effect Effects 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 230000032361 posttranscriptional gene silencing Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000013518 transcription Methods 0.000 description 1
- 230000035897 transcription Effects 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
- 239000013598 vector Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- 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
- C12N15/89—Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation using microinjection
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
- C07K14/43563—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from insects
-
- 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)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Environmental Sciences (AREA)
- Molecular Biology (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Biodiversity & Conservation Biology (AREA)
- Animal Husbandry (AREA)
- Insects & Arthropods (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Animal Behavior & Ethology (AREA)
- Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Microbiology (AREA)
- Toxicology (AREA)
- Plant Pathology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Peptides Or Proteins (AREA)
Abstract
本发明公开了敲除或沉默蜂巢小甲虫气味结合蛋白基因1在防治蜂巢小甲虫中的应用。所述的蜂巢小甲虫气味结合蛋白基因1的核苷酸序列如SEQ ID NO.1所示。本发明采用RNA干扰技术，用dsRNA沉默蜂巢小甲虫OBP1基因的，能影响蜂巢小甲虫对食物源花粉挥发物的选择，可以用于防治蜂巢小甲虫，在蜂巢小甲虫防治过程中具有重要作用。
Description
技术领域
本发明属于昆虫防治领域，具体涉及敲除或沉默蜂巢小甲虫气味结合蛋白基因1在防治蜂巢小甲虫中的应用。
背景技术
蜂巢小甲虫是一种寄生在蜂群内的害虫，原分布于非洲南部，随后扩散至澳洲、美洲、亚洲等各州多个国家，在世界各地均引起不同程度的蜂群危害和经济损失。蜂巢小甲虫在幼虫期和成虫期对蜜蜂造成严重的危害，它们取食花粉、蜂蜜和蜜蜂幼虫，因而会导致蜜蜂幼虫死亡、蜂蜜发酵和巢脾损毁，常造成整个蜂巢坍塌、蜂群弃巢飞逃。目前对其防治主要依靠化学防治，然而长期施用化学杀虫剂导致一系列问题，如环境污染，抗药性产生和对非靶标生物的危害等。因此，开发新型、可替代化学防治的方法变得极其紧迫。
RNA干扰(RNA interference,RNAi)是指在进化过程中高度保守的、由双链RNA(double-stranded RNA，dsRNA)引发的特异性转录后基因沉默现象。由于该技术对靶基因的高效性、特异性及易操作性等特点，已经在基因功能研究、基因治疗药物的开发及作物病虫害的防治方面展示了广阔的应用前景。
发明内容
本发明的目的是提供敲除或沉默蜂巢小甲虫气味结合蛋白基因1在防治蜂巢小甲虫中的应用，所述的蜂巢小甲虫气味结合蛋白基因1的核苷酸序列如SEQ ID NO.1所示。
本发明通过实验发现，注射dsGFP的蜂巢小甲虫对两种花粉挥发物均具有明显的吸引效果，但是沉默OBP1后，蜂巢小甲虫对花粉挥发物的吸引力下降。
因此敲除或沉默蜂巢小甲虫气味结合蛋白基因1可以用于防治蜂巢小甲虫。
优选，所述的防治蜂巢小甲虫是降低蜂巢小甲虫对花粉挥发物的吸引力。
优选，所述的花粉挥发物是亚麻酸乙酯或棕榈酸乙酯。
优选，所述的沉默蜂巢小甲虫气味结合蛋白基因1，用于沉默的dsRNA的序列如SEQID NO.2所示。
本发明采用RNA干扰技术，用dsRNA沉默蜂巢小甲虫OBP1基因的，能影响蜂巢小甲虫对食物源花粉挥发物的选择，可以用于防治蜂巢小甲虫，例如防治蜂巢小甲虫的制剂，在蜂巢小甲虫防治过程中具有重要作用。
附图说明
图1是OBP1基因的沉默效率；
图2是沉默OBP1后小甲虫对花粉挥发物的影响。
具体实施方式
下面结合实施案例对本发明进行进一步的阐述，但不是对本发明的限制。
实施例1：蜂巢小甲虫气味结合蛋白基因1(OBP1)的序列及其dsRNA的获得
1、蜂巢小甲虫气味结合蛋白基因1(OBP1)的序列获得
1)蜂巢小甲虫OBP1基因序列在蜂巢小甲虫转录组数据库的搜索
基于蜂巢小甲虫转录组数据库，采用生物信息学方法对蜂巢小甲虫OBP1基因的序列进行搜索，经过序列分析及比对后，共获得1条蜂巢小甲虫OBP1基因的序列。
2)PCR扩增所需引物设计：
根据上述基因片段，并采用Primer premier 5.0软件设计上游引物GTAAAAATGAAAACATTCGT和下游引物CGGCTAAATGAGCAGCCGTCAT由上海生工生物工程股份有限公司合成。提取蜂巢小甲虫成虫第7天触角的总RNA，采用M-MLV反转录酶(TaKaRa)将所提RNA反转录成第一链cDNA。从而获得PCR反应所需模板。
3)PCR扩增反应
以此作为模板，结合设计上下游引物，PCR扩增获得OBP1基因全长片段，并通过GelExtroaction Kit(Omega)将PCR产物进行纯化，将纯化后的产物克隆到pEASY-T3 Cloningvector(全式金公司)中，转入感受态细胞，扩大菌液培养，采用Plasmid Mini Kit(Omega)提取质粒检测后将菌液送往生工生物工程(上海)股份有限公司进行测序。测序得到核苷酸序列为：
GTAAAAATGAAAACATTCGTGGTTTTATTTTCTCTGTTGGCACTCACAAATGCCGTGTCCAGAGAATTTGTGGAAAATTTTATGAATAAACTACAGGAAGTTGGGGAAAAGTGCGCAGAGGAAACCAATGCAACTGACGACGACGTAGCTGCATTAATTGCCCATACGATGCCCGAATCCCACAATGGAAAATGTATGATTTTGTGTTTTAATGTTGCTTTTCATTTGCAAAAACCTGATGGCACCCCAGATAAAGAAGGTTCAATTGCCTCGTTGGAACCTCTAAAAGCGGACGATCCAGAAATGCACGCCAAAGTTCTGAAAATATTTATGACCTGCGGCCAAAAAACTGCGGTTGATGCAGATCCTTGTATGACGGCTGCTCATTTAGCCG。
即为蜂巢小甲虫气味结合蛋白基因1(OBP1)。
2、蜂巢小甲虫气味结合蛋白基因1(OBP1)dsRNA的合成
1)蜂巢小甲虫气味结合蛋白基因1(OBP1)dsRNA引物的设计
基于本研究所得到蜂巢小甲虫OBP1基因的序列(如SEQ ID NO：1所示)，采用Primer premier 5.0软件设计dsRNA引物，上游引物序列为taatacgactcactatagggACTCACAAATGCCGTGTCC，下游引物序列为taatacgactcactatagggCTTCTTTATCTGGGGTGCCA(小写字母代表T7启动子)，所有引物均由生工生物工程(上海)股份有限公司合成。
2)蜂巢小甲虫气味结合蛋白基因1(OBP1)dsRNA的合成
上述dsRNA引物合成PCR产物，经Gel Extraction Kit(Omega)试剂盒纯化后按照T7RiboMAXTMExpress RNAi System(Promega)试剂盒说明书进行体外转录合成dsRNA。使用NaNoDrop 2000(Thermo scientific)进行定量，使其终浓度达到1μg/μL。保存于-80℃超级低温冰箱备用。
实施例2：注射气味结合蛋白基因1(OBP1)的dsRNA实现蜂巢小甲虫头部OBP1的表达量显著下降
1、特异性dsRNA注射
将1μL(1μg)由SEQ ID NO：2所示的dsRNA(dsOBP1，具体为ACTCACAAATGCCGTGTCCAGAGAATTTGTGGAAAATTTTATGAATAAACTACAGGAAGTTGGGGAAAAGTGCGCAGAGGAAACCAATGCAACTGACGACGACGTAGCTGCATTAATTGCCCATACGATGCCCGAATCCCACAATGGAAAATGTATGATTTTGTGTTTTAATGTTGCTTTTCATTTGCAAAAACCTGATGGCACCCCAGATAAAGAAG)用10μL规格微量注射器注射到蜂巢小甲虫第七日龄幼虫体内，共注射100只。注射相同体积浓度的dsGFP(GTGGAGAGGGTGAAGGTGATGCAACATACGGAAAACTTACCCTTAAATTTATTTGCACTACTGGAAAACTACCTGTTCCATGGCCAACACTTGTCACTACTTTCTCTTATGGTGTTCAATGCTTTTCAAGATACCCAGATCATATGAAACAGCATGACTTTTTCAAGAGTGCCATGCCCGAAGGTTATGTACAGGAAAGAACTATATTTTTCAAAGATGACGGGAACTACAAGACACGTGCTGAAGTCAAGTTTGAAGGTGATACCCTTGTTAATAGAATCGAGTTAAAAGGTATTGATTTTAAAGAAGATGGAAACATTCTTGGACACAAATTGGAATACAACTATAACTCACACAATGTATACATCATGGCAGACAAACAAAAGAATGGAATCAAAGTTAACTTCAAAATTAGACACAACATTGAAGATGGAAGCGTTCAACTAGCAGACCATTATCAACAAAATACTCCAATTGGCGATGGCCCTGTCCTTTTACCAGACAACCATTACCTGTCCACACAATCTGCCC，如SEQ ID NO.3所示)至对照组体内，共注射100只。将注射后蜂巢小甲虫置于28℃恒温生化培养箱中饲养(光照：黑暗时间＝0h:24h，温度28±1℃，湿度80％)，对照组和处理组每天均饲喂新鲜的花粉和糖水。
2、蜂巢小甲虫气味结合蛋白基因1(OBP1)的沉默检测
待蜂巢小甲虫羽化后第七天，分别取注射dsGFP和dsOBP1的成虫各15只，解剖头部，进行总RNA提取，并反转录成第一链cDNA，采用Real-time PCR方法分别检测目的基因(OBP1)和管家基因(GAPDH)的相对表达量，从而对其沉默效率进行计算。结果表明，与对照组比较，注射dsRNA后，处理组OBP1基因表达显著降低(图1)。每组设置3个生物学重复，每个生物学重复5头成虫。
实施例3：抑制蜂巢小甲虫OBP1的表达，显著降低蜂巢小甲虫对花粉挥发物的响应行为
对照组(注射dsGFP)和处理组(注射dsOBP1)蜂巢小甲虫均在人工气候箱饲养，发育到成虫第7天后，进行Y形玻璃管(直接0.5cm，主臂长8cm，分支臂长5cm)嗅觉行为实验。将滤纸片(1cm×1cm)放置在分支臂的最末端，滤纸片上滴加5μL正己烷(对照)或5μL正己烷稀释的花粉挥发物包括亚麻酸乙酯(1mg/mL)或棕榈酸乙酯(0.1mg/mL)。实验保持室内温度25℃进行，使用气泵让气流以0.8L/min的恒定速度先后通过活性炭、加湿瓶、流量计和滤纸片。将单只蜂巢小甲虫放在Y形管主臂口，1min后进入分支臂且至少停留30s记为“选择”，停留在主臂超过2min时，记录“没有选择”。每两只小甲虫换一个新的Y形管，减少残留气味的影响。每组进行3次重复(每次重复11-15只蜂巢小甲虫)。结果发现，对照组注射dsGFP的蜂巢小甲虫对两种花粉挥发物均具有明显的吸引效果，但是沉默OBP1后，蜂巢小甲虫对花粉挥发物的吸引力下降(图2)。
序列表
<110> 广东省科学院动物研究所
<120> 敲除或沉默蜂巢小甲虫气味结合蛋白基因1在防治蜂巢小甲虫中的应用
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 394
<212> DNA
<213> 蜂巢小甲虫(Aethina tumida)
<400> 1
gtaaaaatga aaacattcgt ggttttattt tctctgttgg cactcacaaa tgccgtgtcc 60
agagaatttg tggaaaattt tatgaataaa ctacaggaag ttggggaaaa gtgcgcagag 120
gaaaccaatg caactgacga cgacgtagct gcattaattg cccatacgat gcccgaatcc 180
cacaatggaa aatgtatgat tttgtgtttt aatgttgctt ttcatttgca aaaacctgat 240
ggcaccccag ataaagaagg ttcaattgcc tcgttggaac ctctaaaagc ggacgatcca 300
gaaatgcacg ccaaagttct gaaaatattt atgacctgcg gccaaaaaac tgcggttgat 360
gcagatcctt gtatgacggc tgctcattta gccg 394
<210> 2
<211> 218
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 2
actcacaaat gccgtgtcca gagaatttgt ggaaaatttt atgaataaac tacaggaagt 60
tggggaaaag tgcgcagagg aaaccaatgc aactgacgac gacgtagctg cattaattgc 120
ccatacgatg cccgaatccc acaatggaaa atgtatgatt ttgtgtttta atgttgcttt 180
tcatttgcaa aaacctgatg gcaccccaga taaagaag 218
<210> 3
<211> 531
<212> DNA
<213> 人工序列(Artificial Sequence)
<400> 3
gtggagaggg tgaaggtgat gcaacatacg gaaaacttac ccttaaattt atttgcacta 60
ctggaaaact acctgttcca tggccaacac ttgtcactac tttctcttat ggtgttcaat 120
gcttttcaag atacccagat catatgaaac agcatgactt tttcaagagt gccatgcccg 180
aaggttatgt acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgtg 240
ctgaagtcaa gtttgaaggt gatacccttg ttaatagaat cgagttaaaa ggtattgatt 300
ttaaagaaga tggaaacatt cttggacaca aattggaata caactataac tcacacaatg 360
tatacatcat ggcagacaaa caaaagaatg gaatcaaagt taacttcaaa attagacaca 420
acattgaaga tggaagcgtt caactagcag accattatca acaaaatact ccaattggcg 480
atggccctgt ccttttacca gacaaccatt acctgtccac acaatctgcc c 531
Claims (4)
1.敲除或沉默蜂巢小甲虫气味结合蛋白基因1在防治蜂巢小甲虫中的应用，所述的蜂巢小甲虫气味结合蛋白基因1的核苷酸序列如SEQ ID NO.1所示，所述的防治蜂巢小甲虫是降低蜂巢小甲虫对花粉挥发物的吸引力。
2.根据权利要求1所述的应用，其特征在于，所述的花粉挥发物是亚麻酸乙酯。
3.根据权利要求1所述的应用，其特征在于，所述的花粉挥发物是棕榈酸乙酯。
4.根据权利要求1所述的应用，其特征在于，所述的沉默蜂巢小甲虫气味结合蛋白基因1，用于沉默的dsRNA的序列如SEQ ID NO.2所示。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111183680.1A CN113755530B (zh) | 2021-10-11 | 2021-10-11 | 敲除或沉默蜂巢小甲虫气味结合蛋白基因1在防治蜂巢小甲虫中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202111183680.1A CN113755530B (zh) | 2021-10-11 | 2021-10-11 | 敲除或沉默蜂巢小甲虫气味结合蛋白基因1在防治蜂巢小甲虫中的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113755530A CN113755530A (zh) | 2021-12-07 |
| CN113755530B true CN113755530B (zh) | 2022-05-17 |
Family
ID=78799171
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202111183680.1A Active CN113755530B (zh) | 2021-10-11 | 2021-10-11 | 敲除或沉默蜂巢小甲虫气味结合蛋白基因1在防治蜂巢小甲虫中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113755530B (zh) |
-
2021
- 2021-10-11 CN CN202111183680.1A patent/CN113755530B/zh active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN113755530A (zh) | 2021-12-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110468137B (zh) | 一种茄二十八星瓢虫高致死基因及其在防治瓢虫的应用 | |
| CN110592117B (zh) | 高致死基因vATPase B及其防瓢虫应用 | |
| CN114957426B (zh) | Sp6rars及其在防治蜚蠊目昆虫中的应用 | |
| CN110616223B (zh) | 一种防治茄二十八星瓢虫的靶基因及其应用 | |
| CN110551730B (zh) | 茄二十八星瓢虫rps18基因及其在防虫害中的应用 | |
| CN106676112B (zh) | 桔小实蝇Taiman基因和其qRT-PCR检测方法以及其siRNA | |
| CN113755530B (zh) | 敲除或沉默蜂巢小甲虫气味结合蛋白基因1在防治蜂巢小甲虫中的应用 | |
| CN110616222B (zh) | 防治茄二十八星瓢虫的靶基因及其应用 | |
| CN110564702B (zh) | 二化螟生长发育相关蛋白ND及其编码基因、dsRNA干扰序列和应用 | |
| CN110511936B (zh) | 茄二十八星瓢虫生长发育相关基因chs1及其应用 | |
| CN110144360B (zh) | 一种二化螟SDR基因及其编码的蛋白质和应用、dsRNA及其扩增的引物对和应用 | |
| CN109913480B (zh) | 一种蝗虫尿苷二磷酸葡萄糖醛酸转移酶基因及其应用 | |
| CN110511937B (zh) | Snf7基因及其防治瓢虫的应用 | |
| CN110592044B (zh) | 蛋白激酶Fused编码基因及其在防治小菜蛾中的应用 | |
| CN109112117B (zh) | 一种分离的二化螟cyp15c1基因及其编码蛋白 | |
| CN108277220B (zh) | srp54k基因及其特异性dsRNA在柳蓝叶甲防治中的应用 | |
| CN114854761B (zh) | AtSpook基因在防治蜂巢小甲虫中的应用 | |
| CN110628773B (zh) | 一种用于防治酸浆瓢虫的靶基因和方法 | |
| CN108949769A (zh) | 一种棉铃虫蜕皮激素调控因子E78-C基因cDNA及其应用 | |
| CN110628771B (zh) | 一种防治酸浆瓢虫的试剂盒 | |
| CN115927349B (zh) | 黑翅土白蚁革兰氏阴性菌识别蛋白gnbp2基因及其应用 | |
| CN110628774B (zh) | 基因deltaCOPI及其在防治酸浆瓢虫中的应用 | |
| CN110628772B (zh) | 基因gammaCOPI及其在防治酸浆瓢虫中的应用 | |
| CN118006668A (zh) | 柑橘木虱DcS44基因在防治柑橘木虱及黄龙病中的应用 | |
| CN117431261A (zh) | 一种能提高粘质沙雷氏菌防治白蚁效果的OfDrosha基因dsRNA |
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 |