CN113502347B - 一种用于鉴别辣椒抗感cmv的引物对及其应用 - Google Patents
一种用于鉴别辣椒抗感cmv的引物对及其应用 Download PDFInfo
- Publication number
- CN113502347B CN113502347B CN202110834217.2A CN202110834217A CN113502347B CN 113502347 B CN113502347 B CN 113502347B CN 202110834217 A CN202110834217 A CN 202110834217A CN 113502347 B CN113502347 B CN 113502347B
- Authority
- CN
- China
- Prior art keywords
- cmv
- plant
- capsicum
- primer pair
- identifying
- 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
- 235000002566 Capsicum Nutrition 0.000 title claims abstract description 26
- 239000001390 capsicum minimum Substances 0.000 title claims abstract description 16
- 240000008574 Capsicum frutescens Species 0.000 title claims abstract 5
- 241000196324 Embryophyta Species 0.000 claims abstract description 25
- 201000010099 disease Diseases 0.000 claims abstract description 12
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 12
- 238000000034 method Methods 0.000 claims description 15
- 108091092584 GDNA Proteins 0.000 claims description 6
- 238000006243 chemical reaction Methods 0.000 claims description 6
- 238000000137 annealing Methods 0.000 claims description 4
- 108020004414 DNA Proteins 0.000 claims description 3
- 240000001140 Mimosa pudica Species 0.000 claims description 3
- 235000016462 Mimosa pudica Nutrition 0.000 claims description 3
- 238000004925 denaturation Methods 0.000 claims description 3
- 230000036425 denaturation Effects 0.000 claims description 3
- 238000002264 polyacrylamide gel electrophoresis Methods 0.000 claims description 3
- 238000012257 pre-denaturation Methods 0.000 claims description 3
- 239000000463 material Substances 0.000 abstract description 16
- 108090000623 proteins and genes Proteins 0.000 abstract description 13
- 239000006002 Pepper Substances 0.000 abstract description 10
- 235000016761 Piper aduncum Nutrition 0.000 abstract description 10
- 235000017804 Piper guineense Nutrition 0.000 abstract description 10
- 235000008184 Piper nigrum Nutrition 0.000 abstract description 10
- 239000003550 marker Substances 0.000 abstract description 10
- 238000009395 breeding Methods 0.000 abstract description 6
- 230000001488 breeding effect Effects 0.000 abstract description 6
- 239000003147 molecular marker Substances 0.000 abstract description 4
- 238000012216 screening Methods 0.000 abstract description 4
- 238000009396 hybridization Methods 0.000 abstract description 3
- 244000203593 Piper nigrum Species 0.000 abstract 1
- 241000208293 Capsicum Species 0.000 description 11
- 241000722363 Piper Species 0.000 description 9
- 241000700605 Viruses Species 0.000 description 4
- 238000013461 design Methods 0.000 description 4
- 206010022000 influenza Diseases 0.000 description 4
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 3
- 108091081024 Start codon Proteins 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 208000015181 infectious disease Diseases 0.000 description 3
- 230000002458 infectious effect Effects 0.000 description 3
- 208000024891 symptom Diseases 0.000 description 3
- 238000011144 upstream manufacturing Methods 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000724252 Cucumber mosaic virus Species 0.000 description 2
- 108091027981 Response element Proteins 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000003780 insertion Methods 0.000 description 2
- 230000037431 insertion Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000003753 real-time PCR Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 230000008261 resistance mechanism Effects 0.000 description 2
- 230000003938 response to stress Effects 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 230000028604 virus induced gene silencing Effects 0.000 description 2
- 108091062157 Cis-regulatory element Proteins 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 108700003861 Dominant Genes Proteins 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 101100047461 Rattus norvegicus Trpm8 gene Proteins 0.000 description 1
- 238000012300 Sequence Analysis Methods 0.000 description 1
- 241000723873 Tobacco mosaic virus Species 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 238000000227 grinding Methods 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000006116 polymerization reaction Methods 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 230000009897 systematic effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 230000009261 transgenic effect Effects 0.000 description 1
- 238000004383 yellowing Methods 0.000 description 1
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
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
-
- 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
- C12Q1/6858—Allele-specific amplification
-
- 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
- C12Q2600/13—Plant traits
-
- 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
- C12Q2600/156—Polymorphic or mutational markers
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Genetics & Genomics (AREA)
- General Health & Medical Sciences (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Botany (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明公开了一种用于鉴别辣椒抗感CMV的引物对及其应用。一种用于鉴别辣椒抗感CMV的引物对,正向引物如SEQ ID NO.1所示,反向引物如SEQ ID NO.2所示。利用辣椒基因组DNA对该标记进行PCR扩增;如果有214 bp的分子标记条带,则为抗病植株;如果有220bp的分子标记条带,则为感病植株。利用该标记可以精准高效的筛选抗CMV育种材料,在杂交和回交群体中可高效、准确的筛选携带抗性基因的后代单株,相较于常规抗CMV育种,可显著加快抗CMV育种材料的创制和新品种的选育。
Description
技术领域
本发明属于农业生物技术工程领域,涉及一种用于鉴别辣椒抗感CMV的引物对及其应用。
背景技术
黄瓜花叶病毒(Cucumber mosaic virus,CMV)是世界十大植物病毒之一,可侵染超过100个科1200种植物。刘勇等(2019)调查显示辣椒中检出的33种病毒中,CMV检出率高达20.29%,超越烟草花叶病毒(检出率14.64%)成为危害我国辣椒生产的第一大优势病毒。CMV作为威胁我国辣椒生产的优势病毒,培育抗性品种是防治CMV最经济有效的方法,也是我国辣椒抗病育种的主攻目标。
国内外育种家通过抗性转育,育成了一系列中抗或耐CMV的辣椒品种,但高抗CMV品种稀缺。很多新的CMV病毒株系可突破原有抗性,导致现有品种已无法满足产业需求。鉴定和应用新的抗CMV基因或将多个抗性基因聚合,育成广谱、持久、高抗CMV 的辣椒品种是解决高抗品种稀缺最有效的方法。辣椒抗CMV 基因的鉴定、克隆及抗性机理解析有助于抗性基因的转育应用,一直是辣椒抗CMV 研究的重点和热点。辣椒上已鉴定出30余个抗CMV相关的QTL和基因,除单显性基因Cmr1外,其他抗性基因均未转育应用,根本原因在于抗性基因和抗性机理模糊不清。
发明内容
本发明的目的是针对现有技术的上述不足,提供一种用于鉴别辣椒抗感CMV的引物对。
本发明的另一目的是提供该引物对的应用。
本发明的又一目的是提供一种鉴定辣椒抗CMV的方法。
本发明的目的可通过以下技术方案实现:
一种用于鉴别辣椒抗感CMV的引物对,正向引物如SEQ ID NO.1所示,反向引物如SEQ ID NO.2所示。
本发明所述的引物对在鉴别辣椒抗、感CMV品种中的应用。
本发明所述的引物对在制备鉴别辣椒抗、感CMV品种的试剂中的应用。
一种鉴定辣椒抗CMV的方法,该方法包括使用本发明所述的引物对进行检测的步骤。
作为本发明的一种优选,所述的方法包括利用本发明所述的引物对对辣椒基因组DNA进行PCR扩增;如果扩增产物有214 bp的条带且没有220bp条带,则为抗病植株;如果有220bp的分子标记条带且没有214bp条带,则为感病植株。
作为本发明的一种优选, PCR反应体系15μl,包括7.5μl 2×Taq Master Mix,2μlgDNA模板,正反向引物分别为0.25μl,ddH2O补足至15μl。
作为本发明的一种优选,PCR反应程序为:94℃预变性3 min;94℃变性20 s,55℃退火30 s,72℃延伸40 s,35个循环;72℃延伸7 min。
作为本发明的一种优选,PCR产物利用8%聚丙烯酰胺凝胶电泳检测,如果有214bp的条带的植株为抗病植株;如果有220 bp的条带且没有214bp条带,则为感病植株;二者同时存在为杂合单株。
有益效果:
申请人在辣椒抗源材料C. frutescens cv. PBC688的2号染色体上定位到1个抗CMV主效QTL(qCmr2.1),实时荧光定量PCR和VIGS等技术明确了CA02g19570为qCmr2.1的候选基因;启动子序列分析显示该区域包含光信号、激素应答和逆境胁迫响应等多种顺式作用元件,抗感材料间存在多个响应元件的差异,起始密码子上游277bp处感病材料存在1个6-bp 插入导致该区域内存在3个串联的胁迫响应原件 STRE,而抗病材料PBC688和CM334中只有2个STRE。基于以上研究基础,开发出1个位于启动子区的InDel标记qCmr2.1-P,用于辣椒抗CMV种质材料筛选、在杂交和回交转育CA02g19570的过程中辅助筛选携带抗性基因的后代单株,用于种质资源创制。
该标记位于抗性基因CA02g19570的启动子区域,距离起始密码子只有277 bp左右,与抗性基因连锁极为紧密,基本不会发生重组,对该抗性基因的筛选效率可达到100%。该标记是一个基于PCR技术的共显性标记,基于该标记开发的引物对可应用于苗期抗CMV选择,相对于抗CMV人工接种鉴定和田间鉴定,省时、省力、高效,从而加速抗CMV种质创制和育种进程。
附图说明
图1. CM334、PBC688和G29间qCmr2.1的启动子区6 bp碱基的插入/缺失
图2. 标记qCmr2.1-P 在抗感材料及回交群体中扩增情况
S:感病材料G29;R:抗病材料PBC688;1-60:回交群体BC2 60个单株
具体实施方式
实施例1
课题组前期研究发现实时荧光定量PCR和VIGS等技术明确了CA02g19570为qCmr2.1的候选基因,该基因对CMV具有抗性。抗CMV材料PBC688为母本,感病材料G29为父本,杂交获得F1杂交种,G29为回交父本回交2代获得BC2群体。
分子标记的引物设计: 根据起始密码子上游277 bp处感病材料存在1个6-bp 插入(图1),取该InDel位点两翼各150 bp碱基长度,共300 bp长度用于引物设计。采用Primer5.0 进行引物设计,上游引物设计范围在1~150 bp,下游引物设计范围为 156~300 bp,产物大小为150~250 bp之间,退火温度在52~60 ℃。
分子标记的引物序列为:
qCmr2.1-P-F:AAGGGGGAGTATGATTATTGTT(SEQ ID NO.1);
qCmr2.1-P-R:TATTTATTTACCGCCTACGAGT(SEQ ID NO.2);
利用新型快速植物基因组DNA提取试剂盒提取gDNA(Bioteke, Cat#DP3112),取植株幼嫩叶片0.2 g左右,液氮冷冻研磨,按照试剂盒说明书进行基因组DNA提取。1%琼脂糖凝胶电泳检测gDNA质量,紫外微量分光光度计检测gDNA浓度,调整浓度为20 ng/μL,置于-20℃冰箱保存备用。
PCR反应体系15μl,包括7.5μl 2×Taq Master Mix (Tsingke BiologicalTechnology ) ,2μl gDNA模板,正反向引物分别为0.25μl(100 μmol/L),ddH2O补足至15μl。PCR反应程序为:94℃预变性3 min;94℃变性20 s,55℃退火30 s,72℃延伸40 s,35个循环;72℃延伸7 min。
PCR产物利用8%聚丙烯酰胺凝胶电泳检测,如果有214bp的条带的植株为抗病植株;如果有220 bp的条带且没有214bp条带,则为感病植株;二者同时存在为杂合单株。
为了检测该标记是否可用于杂交和回交群体抗性单株的辅助筛选,本试验应用抗性材料PBC688和感病材料G29 及一个包含60个单株的BC2群体进行验证。结果如图2所示:该标记在抗感材料间具有明显差异,而回交群体基因型与G29相同的为33株,基因型为杂合体的有27株,基本符合1:1的遗传分离比。田间抗CMV鉴定发现27株杂合单株未发现明显CMV症状,33株基因型与母本相同的单株25株表现出明显的系统花叶症状,8株表现出新叶明脉和叶片黄化等轻微症状。以上结果说明该标记引物用于鉴别CMV抗性、感性植株是可行的。
Claims (4)
1.一种鉴定辣椒抗CMV的方法,其特征在于该方法包括使用SEQ ID NO.1和SEQ IDNO.2所示的引物对对辣椒基因组DNA进行PCR扩增;如果扩增产物有214 bp的条带,则为抗病植株;如果有220bp的条带且没有214bp条带,则为感病植株;214 bp和220bp的条带同时存在为杂合单株。
2. 根据权利要求1所述的方法,其特征在于 PCR反应体系15μl,包括7.5μl 2×TaqMaster Mix,2μl gDNA模板,正反向引物分别为0.25μl,ddH2O补足至15μl。
3. 根据权利要求1所述的方法,其特征在于PCR反应程序为:94℃预变性3 min;94℃变性20 s,55℃退火30 s,72℃延伸40 s,35个循环;72℃延伸7 min。
4.根据权利要求1所述的方法,其特征在于PCR产物利用8%聚丙烯酰胺凝胶电泳检测。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110834217.2A CN113502347B (zh) | 2021-07-23 | 2021-07-23 | 一种用于鉴别辣椒抗感cmv的引物对及其应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110834217.2A CN113502347B (zh) | 2021-07-23 | 2021-07-23 | 一种用于鉴别辣椒抗感cmv的引物对及其应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113502347A CN113502347A (zh) | 2021-10-15 |
CN113502347B true CN113502347B (zh) | 2024-05-31 |
Family
ID=78014266
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110834217.2A Active CN113502347B (zh) | 2021-07-23 | 2021-07-23 | 一种用于鉴别辣椒抗感cmv的引物对及其应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN113502347B (zh) |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104560973A (zh) * | 2014-12-24 | 2015-04-29 | 江苏省农业科学院 | 一种获得辣椒疫病抗性候选基因及分子标记的方法及应用 |
CN105296475A (zh) * | 2015-11-06 | 2016-02-03 | 江苏省农业科学院 | 与辣椒抗黄瓜花叶病毒病基因qcmv-2-1连锁的分子标记及其应用 |
CN105368927A (zh) * | 2015-09-16 | 2016-03-02 | 北京市农林科学院 | 与辣椒CMV抗性基因紧密连锁的分子标记genSSR3000及获取、应用 |
KR102142389B1 (ko) * | 2019-05-31 | 2020-08-07 | 서울대학교산학협력단 | Cmv-p1 저항성 고추 품종의 판별을 위한 분자마커 및 이의 용도 |
-
2021
- 2021-07-23 CN CN202110834217.2A patent/CN113502347B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104560973A (zh) * | 2014-12-24 | 2015-04-29 | 江苏省农业科学院 | 一种获得辣椒疫病抗性候选基因及分子标记的方法及应用 |
CN105368927A (zh) * | 2015-09-16 | 2016-03-02 | 北京市农林科学院 | 与辣椒CMV抗性基因紧密连锁的分子标记genSSR3000及获取、应用 |
CN105296475A (zh) * | 2015-11-06 | 2016-02-03 | 江苏省农业科学院 | 与辣椒抗黄瓜花叶病毒病基因qcmv-2-1连锁的分子标记及其应用 |
KR102142389B1 (ko) * | 2019-05-31 | 2020-08-07 | 서울대학교산학협력단 | Cmv-p1 저항성 고추 품종의 판별을 위한 분자마커 및 이의 용도 |
Non-Patent Citations (2)
Title |
---|
Rapid identification of QTLs underlying resistance to Cucumber mosaic virus in pepper (Capsicum frutescens);Guangjun Guo等;Theor Appl Genet;第130卷;摘要和补充文件 * |
辣椒抗黄瓜花叶病毒病候选基因功能鉴定;张根莲等;分子植物育种;第18卷(第17期);第5537-5543页 * |
Also Published As
Publication number | Publication date |
---|---|
CN113502347A (zh) | 2021-10-15 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN108715902B (zh) | 梅花垂枝性状snp分子标记及其应用 | |
CN108411028B (zh) | 水稻耐盐基因skc1基因内特异snp共显性分子标记引物及应用 | |
Jamsari et al. | BAC-derived diagnostic markers for sex determination in asparagus | |
Devran et al. | Development of molecular markers tightly linked to Pvr4 gene in pepper using next-generation sequencing | |
CN108165653B (zh) | 用于鉴定辣椒熟性的InDel分子标记及其应用 | |
CN104630364A (zh) | 抗稻瘟病基因Pi9的特异性CAPS标记Pi9caps及其应用 | |
US20200015444A1 (en) | Methods of creating drought tolerant corn plants using markers linked to cold shock domain-containing proteins and compositions thereof | |
CN113637789A (zh) | 小麦抗条锈病基因YrTD121连锁的KASP分子标记及引物、试剂盒和应用 | |
CN110499389B (zh) | 与烟草抗斑萎病位点rtsw紧密连锁的共显性标记引物、鉴别方法及其应用 | |
CN106498068B (zh) | 一种与烟草tmv抗性基因n紧密连锁的共显性ssr标记及其应用 | |
KR100753676B1 (ko) | 배추과 작물의 품종 구분 및 유용표지인자 개발수단으로서의 왜성전이인자, 이를 이용한 dna 표지인자제작용 프라이머, 및 이들을 이용한 배추과 작물의 품종구분방법 | |
CN111926104B (zh) | 一种鉴定甘蔗与斑茅杂交后代真实性的ssr分子标记及其方法 | |
CN110551843B (zh) | 可区分烟草抗斑萎病位点rtsw纯合杂合基因型的共显性标记引物、区分方法及其应用 | |
CN105543366B (zh) | 水稻抗稻瘟病基因Pi25基因内特异SNP共显性分子标记开发及应用 | |
CN105624327B (zh) | 基于番茄黄化曲叶病毒病抗性基因Ty-3的CAPS分子标记 | |
CN109554494B (zh) | 水稻抗褐飞虱bph9复等位基因的通用共显性分子标记及其检测方法和应用 | |
CN113502347B (zh) | 一种用于鉴别辣椒抗感cmv的引物对及其应用 | |
CN113249509A (zh) | 用于美洲黑杨和小叶杨种间杂交子代的鉴别引物和鉴定方法 | |
CN111575398A (zh) | 玉米第4号染色体雄性不育相关基因的分子标记及其应用 | |
CN109468330A (zh) | 大麦黄花叶病抗性基因eIF4EHOR3298及其鉴定方法和应用 | |
CN111363846B (zh) | 一种检测小麦粒重基因QTkw.saas-2D的分子标记及应用 | |
CN111793706B (zh) | 一种豇豆InDel分子标记检测引物组及试剂盒 | |
CN112575101B (zh) | 与西葫芦prsv-w病毒病抗性相关的分子标记及其应用 | |
CN109234438B (zh) | 与黄瓜无限生长型基因共分离的分子标记Indel-De3 | |
CN111826457B (zh) | 用于鉴定绿豆抗白粉病表型的分子标记snp#2及其引物和应用 |
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 |