CN113462803A - Cucumber whole genome background selection SNP (Single nucleotide polymorphism) marker system based on multiplex PCR (polymerase chain reaction) sequencing and application thereof - Google Patents
Cucumber whole genome background selection SNP (Single nucleotide polymorphism) marker system based on multiplex PCR (polymerase chain reaction) sequencing and application thereof Download PDFInfo
- Publication number
- CN113462803A CN113462803A CN202110880403.XA CN202110880403A CN113462803A CN 113462803 A CN113462803 A CN 113462803A CN 202110880403 A CN202110880403 A CN 202110880403A CN 113462803 A CN113462803 A CN 113462803A
- Authority
- CN
- China
- Prior art keywords
- cucumber
- snp
- snp marker
- dna
- multiplex pcr
- 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.)
- Pending
Links
- 240000008067 Cucumis sativus Species 0.000 title claims abstract description 86
- 235000010799 Cucumis sativus var sativus Nutrition 0.000 title claims abstract description 85
- 239000003550 marker Substances 0.000 title claims abstract description 33
- 238000012163 sequencing technique Methods 0.000 title claims abstract description 21
- 238000007403 mPCR Methods 0.000 title claims abstract description 18
- 239000002773 nucleotide Substances 0.000 title description 4
- 125000003729 nucleotide group Chemical group 0.000 title description 4
- 238000003752 polymerase chain reaction Methods 0.000 title description 3
- 230000002068 genetic effect Effects 0.000 claims abstract description 19
- 238000004458 analytical method Methods 0.000 claims abstract description 17
- 238000001514 detection method Methods 0.000 claims abstract description 14
- 238000005516 engineering process Methods 0.000 claims abstract description 14
- 238000010276 construction Methods 0.000 claims abstract description 5
- 238000000034 method Methods 0.000 claims description 9
- 238000009395 breeding Methods 0.000 claims description 4
- 230000001488 breeding effect Effects 0.000 claims description 4
- 238000012098 association analyses Methods 0.000 claims description 2
- 108090000623 proteins and genes Proteins 0.000 claims description 2
- 238000011144 upstream manufacturing Methods 0.000 claims description 2
- 238000012214 genetic breeding Methods 0.000 abstract description 3
- 238000003766 bioinformatics method Methods 0.000 abstract description 2
- 238000009827 uniform distribution Methods 0.000 abstract description 2
- 108020004414 DNA Proteins 0.000 description 19
- 239000000463 material Substances 0.000 description 11
- 108700028369 Alleles Proteins 0.000 description 6
- 210000000349 chromosome Anatomy 0.000 description 4
- 238000000513 principal component analysis Methods 0.000 description 4
- 238000009826 distribution Methods 0.000 description 3
- 238000012252 genetic analysis Methods 0.000 description 3
- 230000003321 amplification Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 239000011159 matrix material Substances 0.000 description 2
- 239000003147 molecular marker Substances 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000000523 sample Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 1
- 235000009849 Cucumis sativus Nutrition 0.000 description 1
- 241000219104 Cucurbitaceae Species 0.000 description 1
- 238000007400 DNA extraction Methods 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 1
- 108091092878 Microsatellite Proteins 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 238000004422 calculation algorithm Methods 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 238000012217 deletion Methods 0.000 description 1
- 230000037430 deletion Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000012869 ethanol precipitation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000005562 fading Methods 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 238000012268 genome sequencing Methods 0.000 description 1
- 238000003205 genotyping method Methods 0.000 description 1
- 238000012165 high-throughput sequencing Methods 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 239000012264 purified product Substances 0.000 description 1
- 239000002096 quantum dot Substances 0.000 description 1
- 238000013515 script Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 241000894007 species Species 0.000 description 1
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
- 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
- C40—COMBINATORIAL TECHNOLOGY
- C40B—COMBINATORIAL CHEMISTRY; LIBRARIES, e.g. CHEMICAL LIBRARIES
- C40B50/00—Methods of creating libraries, e.g. combinatorial synthesis
- C40B50/06—Biochemical methods, e.g. using enzymes or whole viable microorganisms
-
- 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)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Biochemistry (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- Immunology (AREA)
- Biophysics (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Mycology (AREA)
- Botany (AREA)
- General Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses a cucumber whole genome background selection SNP marker system based on multiplex PCR sequencing and application thereof, and relates to the technical field of genetic breeding. The invention selects 518 single copy SNP markers with high polymorphism, good universality and uniform distribution from a large amount of cucumber germplasm resource genome data by using a bioinformatics method, and the SNP marker system can be used for cucumber DNA fingerprint database construction, germplasm resource identification, genetic relationship analysis, seed purity detection and genetic initial positioning. The reference genome version number of the SNP marker is Cucumber (Chinese Long) v2, the multiplex PCR sequencing technology is utilized to type the SNP marker system, the detection accuracy and resolution are high, high-throughput, low-cost and automatic safety detection is realized, and the SNP marker can be used in molecular genetic breeding work in the future.
Description
Technical Field
The invention belongs to the field of genetic breeding, and relates to a cucumber whole genome background selection SNP marker system based on multiplex PCR sequencing and application thereof.
Background
The Single Nucleotide Polymorphism (SNPs) markers have the characteristics of high density, good stability and strong representativeness in a genome, and can be automatically completed in a large scale without judging according to the sizes of fragments in the genotyping process (Guangqiang, Zhang moon, Xuxianglingong, Sundeliang, Liseiuyan, Linhong, Panliyan, Mayanhua (2008) research progress of DNA molecular markers and several novel molecular marker technologies, Heilongjiang agricultural science: 102-. Compared with repeat sequence polymorphism markers such as microsatellites (SSRs), SNPs have higher genetic stability, and genome resequencing is carried out on different individuals in an intraspecies population under the condition of known species genome, so that the difference among the individuals in the population can be found on the whole genome level, the detection efficiency is higher, and the method is more suitable for large-sample-size detection and analysis, so that the current biologists consider the SNP markers to be a molecular marker technology with a great application prospect. With The continuous progress of genomics research, cucumber high-quality reference genome sequencing is successively completed (Huang, S., Li, R., Zhang, Z.et al. & Ren, Y. (2009). The genome of The cucumber, Cucumis sativus L.Nature genetics,41(12),1275-1281.), researchers developed a large number of SNP markers (3,305,010 SNPs in total) using 115 cucumber core germplasm (Qi, J., Liu, X., Shen, D.et al. & Du, Y. (2013.) A genomic variation map genes antigens into The genomic basis of The cucumber family registration and diversity 1510, 45 (12)). However, how to apply these SNP markers to breeding work efficiently is an urgent problem to be solved. Firstly, identifying the SNP marker of the magnitude by adopting a traditional experimental method is time-consuming and labor-consuming, and adopting a high-throughput sequencing method needs to improve the cost and needs certain big data analysis capability; secondly, at present, not all of the SNP markers show high polymorphism, Linkage Disequilibrium (LD) exists among the markers, the SNP markers within the LD fading distance often show coseparation, and the problems can cause lower efficiency in genetic analysis such as variety identification, genetic relationship analysis, seed purity detection, QTL positioning and the like. Therefore, the development of the SNP markers which are good in universality and high in polymorphism and are uniformly distributed in a genome and not linked is of great significance for genetic analysis of cucumber germplasm resources at high efficiency and low cost and the discovery of excellent gene resources.
Disclosure of Invention
The invention aims to provide a cucumber whole-gene background selection SNP marker system based on multiplex PCR sequencing and application thereof, which can be used for cucumber DNA fingerprint database construction, germplasm resource identification, genetic relationship analysis, seed purity detection and molecular breeding, and has low analysis cost and high efficiency.
In order to achieve the purpose, the invention provides the following scheme:
the invention carries out bioinformatics analysis on SNP markers obtained from 115 core germplasms of cucumber, screens sites which are high in diversity and are uniformly distributed on a chromosome, further eliminates the SNP markers positioned in a linkage disequilibrium decline distance through linkage disequilibrium analysis, and designs universal primers on the upstream and downstream of the SNP markers according to a cucumber reference genome, thereby obtaining the cucumber whole-gene background selection SNP marker system.
The specific information of the chromosome where the cucumber whole genome background selection SNP marker system is located, the physical position of the SNP locus and the nucleotide variation type is shown in Table 1. The physical position of the SNP marker is determined based on Cucumber (Chinese Long) v2 of a reference genome.
The genetic analysis of 161 cucumber commercial varieties by using the cucumber whole genome background selection SNP marker system of the invention shows that all 518 SNP markers are detected in 161 parts of materials, the detection efficiency reaches 100%, each SNP marker can detect 1-2 alleles, the detected minimum allele frequency range is 0-0.497, the average is 0.234, wherein the minimum allele frequency of more than 80% of the markers (415) is more than 0.1, namely, at least more than 10% of the genotypes of the varieties (16) are inconsistent with other varieties, as shown in figure 1. The number of the least different genotypes between the two varieties is 4, and the number of the most different genotypes is 338. Clustering analysis can clearly separate the 161 materials into different groups according to the distance of genetic distance: the analysis of the colony structure shows that the 161 materials have obvious colony structure, as shown in figure 3.
The invention further discloses a primer pair (Table 1) for detecting the cucumber whole genome background selection SNP marker system, and the design principle of the primer is well known to the technical personnel in the field.
The cucumber whole genome background selection SNP marker system can be applied to the construction of a cucumber DNA fingerprint database, and comprises the following steps: (1) extracting genome DNA of all cucumber varieties for constructing a cucumber DNA fingerprint database; (2) and (3) detecting the 518 SNP sites on the genome DNA of each cucumber variety by using the primers and based on a multiplex PCR sequencing technology, thereby constructing a cucumber DNA fingerprint database.
The cucumber whole genome background selection SNP marker system can be applied to cucumber germplasm resource identification, and comprises the following steps: (1) extracting genome DNA of a standard cucumber variety and a variety to be identified; (2) the primers are used for detecting the 518 SNP loci of the genome DNA of each cucumber variety on the basis of a multiplex PCR sequencing technology; (3) if the 518 SNP loci of the standard variety of the cucumber variety to be identified have the same genotype, identifying the standard variety of the cucumber as the same cucumber variety; otherwise, the cucumber is identified as different cucumber varieties.
The cucumber whole genome background selection SNP marker system can be applied to cucumber germplasm genetic relationship analysis and comprises the following steps: (1) extracting genomic DNA for different cucumber varieties; (2) the primers are used for detecting the 518 SNP loci of the genome DNA of each cucumber variety on the basis of a multiplex PCR sequencing technology; (3) and (3) establishing a clustering dendrogram according to the genotype data in the step (2), and analyzing the genetic relationship of different cucumber varieties.
The cucumber whole genome background selection SNP marker system can be applied to cucumber seed purity detection, and comprises the following steps: (1) extracting genome DNA of the same batch of seeds; (2) the primers are used for detecting the 518 SNP loci of the genome DNA of each seed based on a multiplex PC R sequencing technology; (3) and (3) calculating the number of SNPs between every two seeds according to the genotype data in the step (2), thereby calculating the purity of the seeds.
The cucumber whole genome background selection SNP marker system can be applied to cucumber genetic initial positioning and comprises the following steps: (1) obtaining phenotype data of a cucumber population to be detected and extracting genome DNA of the cucumber population to be detected; (2) the primers are used for detecting the 518 SNP loci of the genome DNA of each seed based on a multiplex PCR sequencing technology; (3) and (3) carrying out QTL positioning and whole genome association analysis according to the phenotype data of (1) and the genotype data of (2), finding SNP markers linked with the characters, and realizing initial genetic positioning.
The invention discloses the following technical effects:
the invention discloses a cucumber whole genome background selection SNP marker system based on multiplex PCR sequencing and application thereof, the SNP marker system has high polymorphism, good universality and reasonable distribution on a genome, can be used for cucumber DNA fingerprint database construction, germplasm resource identification, genetic relationship identification, seed purity detection and molecular breeding, and can be typed by utilizing the multiplex PCR sequencing technology, the detection accuracy and resolution are high, and the high-throughput, low-cost and automatic safety detection is realized.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings needed in the embodiments will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.
FIG. 1 is a histogram of the Minimum Allele Frequency (MAF) distribution of 518 markers;
FIG. 2 is a graph of the change in delta k for population structure analysis using the markers of the present invention;
fig. 3 shows the clustering analysis, principal component analysis and population genetic structure analysis of 161 cucumber main cultivated lines by using the markers of the present invention.
Detailed Description
Example 1
Screening for SNP markers
Cucumber SNP typing data (ftp:// www.icugi.org/pub/reseq/cucumber /) are downloaded by a public database, SNP markers with high polymorphism, high universality and uniform distribution on 7 chromosomes of cucumber are selected by using vcttool software and PERL scripts based on a whole genome re-sequencing result of 115 cucumber core germplasm resources, the genotype deletion rate is required to be less than or equal to 0.2, the minimum allele frequency is required to be greater than or equal to 0.35, genotype difference exists between at least more than 30 markers between any two cucumber resources, and the minimum allele frequency distribution is shown in figure 1. The SNP loci on 7 chromosomes were analyzed for linkage disequilibrium by TASSEL software, and SNP markers located within the linkage disequilibrium regression distance were removed, thereby obtaining the SNP markers of the present invention, as shown in Table 1.
2. Material source and DNA extraction
Green leaves of 161 commercial cucumber cultivar materials were selected, Genomic DNA was extracted using the TIANGEN Plant Genomic DN A Kit, and the obtained DNA was subjected to ordinary agarose electrophoresis, Nanodrop, and Qubit, respectively, to determine sample integrity and concentration.
3. Sequencing and SNP typing
Cucumber (Chinese Long) v2 is used as a reference genome, forward and reverse primers (table 1) are designed according to sequences corresponding to SNP loci, sample material DNA is used as a template for multiplex PCR amplification, 2% agarose gel electrophoresis is carried out on amplification products, the PCR products are purified by an ethanol precipitation method, the purified products are subjected to sequence determination, and finally, sequencing results are compared to the reference genome by BWA software for SNP typing.
TABLE 1 cucumber SNP marker information developed by the present invention
4. Genetic relationship analysis
Screening obtained SNP marker basic data, using Cladogram function of TASSEL software, constructing a phylogenetic tree by using a Neighbor Joining (NJ), and processing the NJ tree by using ITOL (http:// itol.embl.de /); performing Principal Component Analysis (PCA) by using EIGENSOFT software, firstly converting SNP data into a characteristic data matrix, then calculating a characteristic vector of the matrix to effectively distinguish a population, and mapping by using an R language; the group Structure (Structure) analysis is performed by classifying subgroups based on genotype information of molecular markers using a bayesian algorithm using a software Structure 2.3.4, presetting a group number (K) to 1 to 7, setting length of non-iteration (length of burn-in period) at the beginning of MCMC (markov chain monte carlo) to 100,000, setting MCMC after non-iteration to 100,000, performing operations under a mixed model and a frequency correlation model independently, performing simulation operations 10 times for each K value using software, and determining an appropriate K value, i.e., the number of subgroups of the group, using the Structure Harvester software according to a posterior probability value output by the Structure software and a change rate of two successive posterior probability values, as shown in fig. 2, and clustering results, principal component analysis, and group genetic Structure analysis results are shown in fig. 3.
5. Germplasm identification by using SNP marker
According to the results of the genotype analysis of the materials in Table 1, the characteristic SNP markers of different materials are analyzed, and the SNP genotype detection of different materials is carried out by using the characteristic SNP markers, so that the characteristic SNP genotype of the material can be obtained, and the material can be distinguished from other materials.
The above-described embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various modifications and improvements of the technical solutions of the present invention can be made by those skilled in the art without departing from the spirit of the present invention, and the technical solutions of the present invention are within the scope of the present invention defined by the claims.
Claims (9)
1. Cucumber whole genome background selection SNP marker system, characterized in that the locus information of the SNP marker in cucumber is as follows:
2. the cucumber whole genome background selection SNP marker system according to claim 1, wherein the reference genome sequence version number of the SNP marker is Cucumber (Chinese Long) v 2.
3. The primer sequence and the sequence of 150bp each upstream and downstream of the SNP marker according to claim 1 or 2.
4. The cucumber whole gene background selection SNP marker system as set forth in claim 1 or 2, wherein the SNP marker system is applied to cucumber DNA fingerprint database construction, germplasm resource identification, genetic relationship analysis, seed purity detection and molecular breeding.
5. A method for constructing a cucumber DNA fingerprint database is characterized by comprising the following steps:
(1) extracting genome DNA of all cucumber varieties for constructing a cucumber DNA fingerprint database;
(2) and (2) detecting the 518 SNP sites on the genome DNA of each cucumber variety by using the primers as claimed in claim 3 based on a multiplex PCR sequencing technology, thereby constructing a cucumber DNA fingerprint database.
6. A method for identifying cucumber germplasm resources is characterized by comprising the following steps:
(1) extracting genome DNA of a standard cucumber variety and a variety to be identified;
(2) detecting the 518 SNP sites on the genomic DNA of each cucumber variety based on a multiplex PCR sequencing technology by using the primers of claim 3;
(3) if the 518 SNP loci of the standard variety of the cucumber variety to be identified have the same genotype, identifying the standard variety of the cucumber as the same cucumber variety; otherwise, the cucumber is identified as different cucumber varieties.
7. A method for analyzing genetic relationship of cucumber germplasm is characterized by comprising the following steps:
(1) extracting genomic DNA for different cucumber varieties;
(2) detecting the 518 SNP sites on the genomic DNA of each cucumber variety based on a multiplex PCR sequencing technology by using the primers of claim 3;
(3) and (3) establishing a clustering dendrogram according to the genotype data in the step (2), and analyzing the genetic relationship of different cucumber varieties.
8. A method for detecting purity of cucumber seeds is characterized by comprising the following steps:
(1) extracting genome DNA of the same batch of seeds;
(2) detecting the 518 SNP sites on the genomic DNA of each seed based on a multiplex PCR sequencing technology by using the primers of claim 3;
(3) and (3) calculating the number of SNPs between every two seeds according to the genotype data in the step (2), thereby calculating the purity of the seeds.
9. A cucumber genetic initial positioning method is characterized by comprising the following steps:
(1) obtaining phenotype data of a cucumber population to be detected and extracting genome DNA of the cucumber population to be detected;
(2) detecting the 518 SNP sites on the genomic DNA of each seed based on a multiplex PCR sequencing technology by using the primers of claim 3;
(3) and (3) carrying out QTL positioning and whole genome association analysis according to the phenotype data of (1) and the genotype data of (2), finding SNP markers linked with the characters, and realizing initial genetic positioning.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110880403.XA CN113462803A (en) | 2021-08-02 | 2021-08-02 | Cucumber whole genome background selection SNP (Single nucleotide polymorphism) marker system based on multiplex PCR (polymerase chain reaction) sequencing and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110880403.XA CN113462803A (en) | 2021-08-02 | 2021-08-02 | Cucumber whole genome background selection SNP (Single nucleotide polymorphism) marker system based on multiplex PCR (polymerase chain reaction) sequencing and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113462803A true CN113462803A (en) | 2021-10-01 |
Family
ID=77883629
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110880403.XA Pending CN113462803A (en) | 2021-08-02 | 2021-08-02 | Cucumber whole genome background selection SNP (Single nucleotide polymorphism) marker system based on multiplex PCR (polymerase chain reaction) sequencing and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113462803A (en) |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105886602A (en) * | 2014-12-01 | 2016-08-24 | 南京农业大学 | Cucumber mitochondrial genome SSR marker development and application of marker in seed purity identification |
| CN106916879A (en) * | 2015-12-25 | 2017-07-04 | 南京农业大学 | Mitochondrial genomes polymorphism builds cucumber finger-print and its application |
| US20200407805A1 (en) * | 2017-05-05 | 2020-12-31 | Tianjin Kernel Agricultural Science And Technology Corporation Ltd. Cucumber Research Institute | Cucumber Male Sterility Gene, Molecular Marker, Screening Method and Application Thereof |
-
2021
- 2021-08-02 CN CN202110880403.XA patent/CN113462803A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105886602A (en) * | 2014-12-01 | 2016-08-24 | 南京农业大学 | Cucumber mitochondrial genome SSR marker development and application of marker in seed purity identification |
| CN106916879A (en) * | 2015-12-25 | 2017-07-04 | 南京农业大学 | Mitochondrial genomes polymorphism builds cucumber finger-print and its application |
| US20200407805A1 (en) * | 2017-05-05 | 2020-12-31 | Tianjin Kernel Agricultural Science And Technology Corporation Ltd. Cucumber Research Institute | Cucumber Male Sterility Gene, Molecular Marker, Screening Method and Application Thereof |
Non-Patent Citations (4)
| Title |
|---|
| JIAN ZHANG 等: "A new Snp genotyping technology target Snp-seq and its application in genetic analysis of cucumber varieties.", SCIENTIFIC REPORTS * |
| 姚丹青;楼坚锋;朱文莹;张微微;夏建明;: "基于SNP标记的黄瓜遗传多样性分析", 上海农业学报 * |
| 孙振久;王亚娟;张显;: "黄瓜分子标记辅助育种研究进展", 西北植物学报 * |
| 李斯更;沈镝;刘博;邱杨;张晓辉;张忠华;王海平;李锡香;: "基于黄瓜基因组重测序的InDel标记开发及其应用", 植物遗传资源学报 * |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Huang et al. | High-throughput genotyping by whole-genome resequencing | |
| CN108779459B (en) | Cotton whole genome SNP chip and application thereof | |
| US20210285063A1 (en) | Genome-wide maize snp array and use thereof | |
| CN115807122A (en) | SNP molecular marker for pineapple seed resource identification and application thereof | |
| CN108642201B (en) | SNP (Single nucleotide polymorphism) marker related to millet plant height character as well as detection primer and application thereof | |
| CN114480704A (en) | SNP combined marker for identifying eggplant seed resources | |
| CN116926234B (en) | SNP molecular marker related to soybean kernel oil content and application thereof | |
| CN115679012B (en) | Chilli whole genome SNP-Panel and application thereof | |
| CN117133354A (en) | Method for efficiently identifying key breeding gene modules of forest tree | |
| CN112226529A (en) | SNP molecular marker of wax gourd blight-resistant gene and application | |
| CN113249509A (en) | Identification primer and identification method for interspecific hybrid progeny of populus tremuloides and populus tremula | |
| CN108416189B (en) | Crop variety heterosis mode identification method based on molecular marker technology | |
| CN113789407B (en) | SNP molecular marker combination for cyperus esculentus genotyping and application thereof | |
| CN113462803A (en) | Cucumber whole genome background selection SNP (Single nucleotide polymorphism) marker system based on multiplex PCR (polymerase chain reaction) sequencing and application thereof | |
| CN109593871A (en) | Corn KASP molecular labeling for distinguishing Heterosis of Maize Hybrid group combines and its development approach and application | |
| CN115141893A (en) | Molecular marker group containing 7 molecular markers and used for predicting dry matter content of kiwi fruit, application and kit thereof | |
| CN108642203B (en) | SNP (Single nucleotide polymorphism) marker related to millet stem thickness character as well as detection primer and application thereof | |
| CN116479164B (en) | SNP locus, molecular marker, amplification primer and application of SNP locus and molecular marker related to soybean hundred-grain weight and size | |
| CN117248061B (en) | InDel locus related to soybean seed oil content, molecular marker, primer and application thereof | |
| CN117230240B (en) | InDel locus related to soybean seed oil content, molecular marker, primer and application thereof | |
| CN113005215B (en) | Haplotype molecular marker related to poplar wood yield and application thereof | |
| CN116732230B (en) | Rice nitrogen efficient InDel molecular marker GRF4M1 and application thereof | |
| CN117965787B (en) | SNP (Single nucleotide polymorphism) marker and primer set for identifying authenticity of pineapple Josapine and MD2 hybrid and application of SNP marker and primer set | |
| CN108642202B (en) | SNP (Single nucleotide polymorphism) marker related to millet stem node number traits as well as detection primer and application thereof | |
| CN108642197B (en) | SNP (Single nucleotide polymorphism) marker related to millet code number character as well as detection primer 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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20211001 |