CN113981124B - Sakura SSR molecular marker primer and application thereof in identification of 42 sakura varieties - Google Patents
Sakura SSR molecular marker primer and application thereof in identification of 42 sakura varieties Download PDFInfo
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- 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
Abstract
The invention discloses a sakura SSR molecular marker primer and application thereof in 42 sakura variety identification, wherein the invention independently develops the sakura variety SSR related molecular marker primer from tail She Ying genome data, and performs amplification and polymorphism screening on 39 pairs of common SSR marker total primers in the genus prune screened in the literature, performs genetic diversity research on the common 42 sakura varieties, screens out 16 pairs of primers, selects 2 pairs of primers PTCR22 and SC7YT with the highest discrimination rate from the 16 pairs of primers, and can identify 39 types of the 42 sakura varieties, wherein the discrimination rate can reach 92.9%, and then any one of the other 14 pairs of primers is arbitrarily combined, so that all samples can be distinguished. The optimal variety identification primer combination PTCR22+SC7YT+PS12A02 is also screened out, and a sakura variety SSR characteristic fingerprint is constructed, so that an important basis is provided for sakura variety identification and molecular marker assisted breeding.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a sakura SSR molecular marker primer and application thereof in identification of 42 sakura varieties.
Background
Sakura is a generic term for plants of the genus prune (Prunus sensu lato) of the family Rosaceae (Rosaceae). The sakura is a famous world tree, flowers are full of trees when fully bloom, and flowers fall into the flowers and are colorful when the flowers are not full; the plant material has wide development and utilization prospect as fruit trees with higher economic value and elegant garden flowers and trees. Over 600 sakura gardening varieties have been cultivated in recent centuries through natural variation screening and artificial hybridization. However, since many varieties have high morphological similarity, the variety sources are not recorded in detail, and the homonym of the homonym frequently occur; most of the sakura flowers are first flowers and then leaves, and flowers and leaves are in different periods, so that hybridization is easy, morphological variation is abundant and various, life history is long, flowering period is relatively short, and a rapid and accurate identification method is lacked (Wang Xianrong, 2014, shi et al, 2013). Over 600 sakura gardening varieties have been obtained for centuries by means of natural variation screening and hybrid cultivation (Yuan Dongming et al, 2018). For these horticultural varieties, taxonomies sequentially propose three-level, five-level classification standards and flower-period, flower-color classification standards according to morphological characteristics of tree, flower, fruit, leaf, winter bud and the like (Shi Yu, 2007, rattan, jun male, 2009, zhang Qiong and the like, 2012). Because different scholars have different judging standards on morphological characters, the characters between varieties are crossed and overlapped and have strong plasticity in different environments, so that morphological identification is difficult, and the phenomena of variety name confusion, homonym, foreign matter homonym and the like are frequently caused.
Genetic diversity of plant varieties is mainly reflected by the polymorphism of genetic markers, which are genetic markers based on the polymorphism of nucleic acids. The method is not interfered by tissue types, development period, environmental conditions and the like, has the advantages of extremely large quantity, high polymorphism and the like, and is an ideal genetic marker. Molecular markers have been widely used in germplasm resource research, hybrid identification, genetic map construction, gene location of interest, genetic diversity, genetic relationship research, and marker-assisted selection breeding. Wherein, microsatellites (mirosatolites), i.e. simple repeats (simple sequence repeat, SSRs), are simple repeats uniformly distributed in the genome consisting of 2-6 nucleotide tandem repeats; has the advantages of high polymorphism, good stability, multiple alleles, co-dominant property, abundant quantity, good genome coverage, simple operation and the like. The method is applied to researches of genetic relationship, genetic diversity analysis, variety fingerprint drawing, variety identification, DNA fingerprint construction and the like of plants such as Chinese plums (Prunus salicina), peaches (Prunus persica), lycium ruthenicum (Lycium ruthenicum), loquat (Eriobotrya japonica) and the like. The early stage of the research group successfully identifies 20 sakura varieties by using 2 pairs of SSR molecular markers developed from the tail She Ying genome data, and can identify 20 sakura varieties by using only two primers, so that great progress is made, but the sakura varieties are various, and due to the fact that the number of the adopted primers is small, the resolution ratio is relatively insufficient, and part of sakura varieties can not be distinguished. Therefore, the invention further develops the molecular marker primer on the basis of early-stage research, and lays a foundation for identifying more sakura varieties. And carrying out genetic diversity research on the collected common 42 sakura variety materials, and also continuously utilizing SSR markers developed in tail She Ying genome data and partially collecting and screening polymorphic primers to identify the 42 sakura varieties, screening the optimal variety identification primer combination, constructing sakura variety SSR characteristic fingerprints, and providing important basis for sakura variety identification and molecular marker assisted breeding.
Disclosure of Invention
The invention aims at providing a sakura SSR molecular marker primer.
The second purpose of the invention is to provide the effect of the sakura SSR molecular marker primer in sakura variety identification.
The invention further aims at providing a kit for identifying sakura.
In order to achieve the above purpose, the present invention provides the following technical solutions:
the sakura SSR molecular marker primer is characterized in that the molecular marker primer is at least 3 pairs of primer combinations, specifically any 1-14 pairs of PTCR22, SC7YT and ZJF01, ZJF02, ZJF03, BPPCT008, BPPCT040, CPSCT021, EMPaS11A, M a, MA016b, PS12A02, MA007a, pceGA25, UDP97-403, UDP96-005, PTCR22 and SC7YT, and the nucleotide sequences of the forward primer and the reverse primer are specifically as follows:
(1) ZJF01: the forward primer sequence is shown as SEQ ID NO.1, and the reverse primer sequence is shown as SEQ ID NO. 2;
(2) ZJF02: the forward primer sequence is shown as SEQ ID NO.3, and the reverse primer sequence is shown as SEQ ID NO. 4;
(3) ZJF03: the forward primer sequence is shown as SEQ ID NO.5, and the reverse primer sequence is shown as SEQ ID NO. 6;
(4) BPPCT008: the forward primer sequence is shown as SEQ ID NO.7, and the reverse primer sequence is shown as SEQ ID NO. 8;
(5) BPPCT040: the forward primer sequence is shown as SEQ ID NO.9, and the reverse primer sequence is shown as SEQ ID NO. 10;
(6) CPSCT021: the forward primer sequence is shown as SEQ ID NO.11, and the reverse primer sequence is shown as SEQ ID NO. 12;
(7) EMPaS11A: the forward primer sequence is shown as SEQ ID NO.13, and the reverse primer sequence is shown as SEQ ID NO. 14;
(8) M9a: the forward primer sequence is shown as SEQ ID NO.15, and the reverse primer sequence is shown as SEQ ID NO. 16;
(9) MA016b: the forward primer sequence is shown as SEQ ID NO.17, and the reverse primer sequence is shown as SEQ ID NO. 18;
(10) PS12a02: the forward primer sequence is shown as SEQ ID NO.19, and the reverse primer sequence is shown as SEQ ID NO. 20;
(11) MA007a: the forward primer sequence is shown as SEQ ID NO.21, and the reverse primer sequence is shown as SEQ ID NO. 22;
(12) PceGA25: the forward primer sequence is shown as SEQ ID NO.23, and the reverse primer sequence is shown as SEQ ID NO. 24;
(13) UDP97-403: the forward primer sequence is shown as SEQ ID NO.25, and the reverse primer sequence is shown as SEQ ID NO. 26;
(14) UDP96-005: the forward primer sequence is shown as SEQ ID NO.27, and the reverse primer sequence is shown as SEQ ID NO. 28;
(15) PTCR22: the forward primer sequence is shown as SEQ ID NO.29, and the reverse primer sequence is shown as SEQ ID NO. 30;
(16) SC7YT: the forward primer sequence is shown as SEQ ID NO.31, and the reverse primer sequence is shown as SEQ ID NO. 32.
The invention provides application of the sakura SSR molecular marker primer in sakura variety identification, which is used for amplifying 16 pairs of SSR primers issued and 39 pairs of primers in total of common SSR markers of the genus prune screened in the literature and screening polymorphism. Finally, 16 pairs of primers can be screened to distinguish and identify 42 varieties, and the selected sakura varieties are specifically shown in table 1.
2 pairs of primers PTCR22 and SC7YT with highest discrimination rate are selected from 16 pairs of primers, 39 varieties of 42 sakura varieties can be identified by the combination, and the discrimination rate can reach 92.9%. Wherein, tail She Ying fails to amplify the band, and the P17 Pink cherry and the P41 mountain cherry cannot be distinguished. Any one of the remaining 14 pairs of primers is then arbitrarily combined to distinguish between all samples. Considering the amplification of tail She Ying in combination with the polymorphism of each primer, the optimal combination is PTCR22+ SC7YT + PS12A02.
TABLE 1 information on 42 sakura variety
A kit for identifying sakura varieties, comprising SSR molecular marker primers of any 1-14 pairs of PTCR22, SC7YT and 14, ZJF, ZJF, ZJF03, BPPCT008, BPPCT040, CPSCT021, EMPaS11A, M a, MA016b, PS12a02, MA007a, pceGA25, UDP97-403, UDP96-005, PTCR22, SC7YT in combination.
The beneficial effects of the invention are as follows:
according to the invention, through independently developing molecular marker primers related to sakura SSR from tail She Ying genome data and carrying out amplification and polymorphism screening on 39 pairs of primers in total for common SSR markers of plums screened in literature, carrying out genetic diversity research on common 42 sakura varieties, screening out 16 pairs of primers, selecting 2 pairs of primers PTCR22 and SC7YT with highest discrimination rate from the 16 pairs of primers, and combining, 39 kinds of sakura varieties can be identified, the discrimination rate can reach 92.9%, and then any one of the other 14 pairs of primers is combined at will, so that all samples can be discriminated. The optimal variety identification primer combination PTCR22+SC7YT+PS12A02 is also screened out, and a sakura variety SSR characteristic fingerprint is constructed, so that an important basis is provided for sakura variety identification and molecular marker assisted breeding. Enriches the types of molecular marker identification of sakura variety and improves the identification efficiency.
Drawings
Fig. 1 is a cluster dendrogram of 42 sakura varieties constructed based on 16 pairs of SSR markers.
Detailed Description
The principles and features of the present invention are described below with reference to the drawings, the examples are illustrated for the purpose of illustrating the invention and are not to be construed as limiting the scope of the invention.
The technical means used in the examples are conventional means well known to those skilled in the art unless otherwise indicated. The test methods in the following examples are conventional methods unless otherwise specified. The reagents and materials employed, unless otherwise indicated, are commercially available.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In addition, any methods and materials similar or equivalent to those described herein can be used in the present invention. The preferred methods and materials described herein are presented for illustrative purposes only.
1 materials and methods
1.1 test fields and test materials
The experimental plot is set in sakura nursery of the national institute of forestry science, zhejiang, 30 ° 13'12 "north latitude, 120 ° 01' 11" east longitude. The ground belongs to subtropical monsoon climate areas, the four seasons are clear, the annual temperature is moderate, the annual average air temperature is 15.9-17.0 ℃, the extreme highest air temperature is 39.8-42.9 ℃, and the extreme lowest air temperature is-7.1 to-15.0 ℃; the illumination is sufficient, and the average sunshine hours per year is 1 to 2100 hours; air is moist, and the annual average relative humidity is 76% -81%; the rainfall is abundant, and the annual average rainfall is 980-2 000mm; and the frost-free period is 199-328 d.
In 12 months of 2015, the sakura variety was harvested from the sakura breeding bases of 7 provinces of Fujian province, guizhou province, shandong province, hubei province, shanghai city, yunnan province and Zhejiang province and planted in sakura breeding nursery (see Table 1) of the Zhejiang province forestry science institute. Collecting DNA samples of the varieties, respectively placing fresh and disease-free young leaves into self-sealing bags filled with silica gel for rapid drying, and then storing in a freezer at-20 ℃ for standby.
1.2 DNA extraction method
208 parts of genomic DNA of the test material was extracted using a rapid plant genomic DNA extraction kit (model DP 3112) manufactured by Bioteke company, and the sample was pre-treated with a buffer before extraction to remove a large amount of impurities such as polysaccharide, pigment, etc. Detecting DNA quality by 1% agarose gel electrophoresis, measuring DNA concentration by Nanodrop ultra-micro spectrophotometer, and diluting to 50ng.mu.L -1 Placing in a refrigerator at-20deg.C for use.
1.3 SSR primer development
Based on the results of the early subject group sequencing of the tail She Yingquan genome, SSR Primer development was performed using Primer 5.0 software. The 16 pairs of SSR primers and 23 pairs of common SSR markers of the plum genus screened in the literature are amplified and screened for polymorphism together with 39 pairs of primers. Finally, 16 pairs of primers were screened for 42 varieties of identification (SSR primer information is shown in Table 2). Primers were synthesized by the division of biological engineering (Shanghai).
TABLE 2 SSR primer information
1.4 PCR amplification
The PCR amplification system was 25. Mu.L: 50 ng/. Mu.L -1 DNA template 2. Mu.L, 2X TSINGKE Master MIX (blue) 12.5. Mu.L (produced by Beijing engine biosciences Co., ltd.), upstream primer 0.5. Mu.L, downstream primer 0.5. Mu.L, ddH 2 O9.5. Mu.L. The PCR reaction was performed on a Applied Biosystem Veriti Thermal Cycler PCR (hong Kong Gene Company Limited Gene Co., ltd.) instrument, and the reaction procedure was as follows: 95 ℃ for 5min; 45s at 95 ℃, 45s at 55-58 ℃, 45s at 72 ℃ and 35 cycles in total; 7min at 72 ℃; finally, the mixture is preserved at the temperature of 4 ℃. The PCR amplified products were screened for clear bands by 2% agarose gel electrophoresis.
1.5 capillary electrophoresis
1. Mu.L of the PCR product with clear and stable bands was pipetted through Qsep100 TM Full-automatic nucleic acid protein analyzerAnd (5) detecting by capillary electrophoresis. The cartridge used for capillary electrophoresis was an S1 high resolution clamp, and the molecular Size standard was 1K Size marker, both manufactured by Guangzhou Jiyuan biosciences Inc.
1.6 data analysis
And comparing and analyzing the peak values of all varieties output by the full-automatic nucleic acid protein analyzer to obtain the fragment size, and using GeneMarker 2.2.0 software for data analysis. Sequence length data was converted to genotype using the overt software. Calculation of SSR primer allele factors (number of alleles, na), observations of heterozygosity (observed heterozygosity, H o ) Nei's genetic diversity (Nei's gene diversity, H), shannon's informative index (Shannon's information index, I) and polymorphism informative content (polymorphism information content, PIC), were analyzed statistically using PopGene 32 software. The fragment size values are converted into 0 and 1 data matrixes by using MG software, genetic similarity coefficients (coeffients) among samples are calculated by using Ntsys 2.1 software, and clustering is carried out by using a UPGMA method.
2 results
2.1 molecular marker polymorphism analysis
Detecting the extracted DNA by using a Nanodrop ultra-micro spectrophotometer and 1.0% agarose gel electrophoresis, wherein A260/A280 of leaf extracts of 24 sakura varieties is 1.8-2.0, A260/A230 is more than 2.0, and the concentration range of the DNA is 230ng mu L -1 -550ng·μL -1 The method shows that the content of proteins, pigments, phenolic substances and the like in the extracted DNA sample is low, the purity of the DNA sample is high, and the subsequent detection requirements can be met. Randomly selecting 6 varieties of sakura, early pine, silver river mountain sakura, large fish sakura and large cold sakura, and carrying out polymorphism screening on the developed 16 pairs of SSR primers and 39 pairs of SSR primers screened in the literature to obtain 16 pairs of polymorphic primers with clear, stable and good amplified bands (Table 2).
The 16 pairs of SSR primers are used for amplifying and carrying out capillary electrophoresis on genome DNA of 42 sakura varieties respectively to obtain 8-36 polymorphic alleles detected by different primers, 17.5 polymorphic alleles can be detected by each pair of primers on average, and the average effective alleles is 9.21. Wherein, the primer SC7YT has the largest polymorphic allele factors and effective allele numbers amplified for all varieties, which indicates that the primer can better reflect the difference of different varieties. The degree of homozygosity and heterozygosity observed for the 16 pairs of primers are 0.02-0.98, the degree of homozygosity is desirably 0.03-0.37, and the degree of heterozygosity is desirably 0.63-0.97. Shannon's information index is 1.23-3.38, and average is 2.30; the Nei's genetic diversity index was 0.63-0.96, with ZJF being the highest for both shannon diversity index and Nei's genetic diversity (Table 3).
TABLE 3 16 genetic information for SSR markers in 42 sakura varieties
2.2 degree of differentiation of molecular markers on sakura variety
According to the genotype analysis of each variety of sakura, the varieties which can be distinguished by 16 pairs of SSR primers are different from 4-30, and the average number is 13.12. Wherein, the number of varieties distinguished by M9a and MA016b is minimum, the distinguishing rate is 9.52%, the primer M9a can distinguish 4 varieties of No. Zhong Huaying excellent plant 4, cercis chinensis, ainsliaea sinensis and Yupalace field cherry, and the primer MA016b can distinguish 4 varieties of Cercis chinensis, chinese red cherry, pinus koraiensis and Shansheng cherry. The discrimination rate of the primer SC7YT is highest and reaches 71.43%, and 30 varieties can be distinguished; the second is the primers PTCR22 and ZJF03, the distinguishing rate is 69.05% and 61.90%, and 29 varieties and 26 varieties can be distinguished.
TABLE 4 differentiation of sakura variety by molecular markers
2.3 SSR marker-based sakura variety genetic relationship analysis
The genetic similarity coefficient of 42 sakura varieties ranges from 0.80 to 0.94, wherein the sakura and the tail She Ying have the lowest similarity, which indicates that the relatives of the two varieties are farthest. The similarity of the cherokee rose, the cherokee rose and the American cherokee rose is highest; among them, green cherokee rose is a variety bred in the cherokee rose, and American cherokee rose and dyeing well Ji Ye come from the same parent and parent, which proves to a certain extent that 16 pairs of primers selected by us can correctly reflect the genetic relationship among varieties. When the genetic similarity coefficient is 0.852, the 42 sakura varieties are gathered together; when the genetic similarity coefficient was 0.94, 42 varieties of sakura were grouped into 42 groups, which were completely distinguishable (fig. 1).
2.4 screening of optimal primer combinations
2 pairs of primers PTCR22 and SC7YT with highest discrimination rate are selected from 16 pairs of primers, 39 varieties of 42 sakura varieties can be identified by the combination, and the discrimination rate can reach 92.9%. Wherein, tail She Ying fails to amplify the band, and the P17 Pink cherry and the P41 mountain cherry cannot be distinguished. Any one of the remaining 14 pairs of primers is then arbitrarily combined to distinguish between all samples. Considering the amplification of tail She Ying and the polymorphism of each primer, the optimal combination of these 42 sakura varieties was identified as ptcr22+sc7yt+ps12a02.
2.5 SSR characteristic fingerprint and variety identification of sakura variety
42 different genotypes were detected in total in 42 sakura varieties based on the primer combination ptcr22+sc7yt+ps12a02. Based on the 3 pairs of primers and the genotypes corresponding to each variety, the (primer_genotype/primer_genotype) combination is used as SSR characteristic fingerprints of 42 sakura varieties, and particularly referring to Table 5, a molecular basis is provided for identifying each sakura variety.
TABLE 5 SSR characteristic fingerprints (primer+genotype) for 42 sakura varieties
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.
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<212> DNA
<213> Prunus dielsiana
<400> 11
gccacttcgg ctaaaagaga 20
<210> 12
<211> 21
<212> DNA
<213> Prunus dielsiana
<400> 12
tccatatctc ctcctgcttg a 21
<210> 13
<211> 20
<212> DNA
<213> Prunus dielsiana
<400> 13
accactttga ggaacttggg 20
<210> 14
<211> 20
<212> DNA
<213> Prunus dielsiana
<400> 14
ctgcctggaa gagcaataac 20
<210> 15
<211> 20
<212> DNA
<213> Prunus dielsiana
<400> 15
gccgaaaccc taggtgagcg 20
<210> 16
<211> 20
<212> DNA
<213> Prunus dielsiana
<400> 16
caggatgctt gcggtgcttg 20
<210> 17
<211> 20
<212> DNA
<213> Prunus dielsiana
<400> 17
tggctggtgg agacggagga 20
<210> 18
<211> 20
<212> DNA
<213> Prunus dielsiana
<400> 18
atgataccca gcctcccggg 20
<210> 19
<211> 18
<212> DNA
<213> Prunus dielsiana
<400> 19
gccaccaatg gttcttcc 18
<210> 20
<211> 18
<212> DNA
<213> Prunus dielsiana
<400> 20
agcaccagat gcacctga 18
<210> 21
<211> 20
<212> DNA
<213> Prunus dielsiana
<400> 21
gtgcatcgtt aggaactgcc 20
<210> 22
<211> 20
<212> DNA
<213> Prunus dielsiana
<400> 22
gcccctgaga tacaactgca 20
<210> 23
<211> 24
<212> DNA
<213> Prunus dielsiana
<400> 23
gcaattcgag ctgtatttca gatg 24
<210> 24
<211> 24
<212> DNA
<213> Prunus dielsiana
<400> 24
cagttggcgg ctatcatgtc ttac 24
<210> 25
<211> 20
<212> DNA
<213> Prunus dielsiana
<400> 25
ctggcttaca actcgcaagc 20
<210> 26
<211> 20
<212> DNA
<213> Prunus dielsiana
<400> 26
cgtcgaccaa ctgagactca 20
<210> 27
<211> 20
<212> DNA
<213> Prunus dielsiana
<400> 27
gtaacgctcg ctaccacaaa 20
<210> 28
<211> 19
<212> DNA
<213> Prunus dielsiana
<400> 28
cctgcatatc accacccag 19
<210> 29
<211> 21
<212> DNA
<213> Prunus dielsiana
<400> 29
tgtgctaatg ccaaaaatac c 21
<210> 30
<211> 20
<212> DNA
<213> Prunus dielsiana
<400> 30
acatgcattt caacccactc 20
<210> 31
<211> 19
<212> DNA
<213> Prunus dielsiana
<400> 31
tgcatgagaa acttgtggc 19
<210> 32
<211> 19
<212> DNA
<213> Prunus dielsiana
<400> 32
tgcatgagaa acttgtggc 19
Claims (2)
1. Application of sakura SSR molecular marker primer combination in 42 sakura variety identification is characterized in that the molecular marker primer combination is PTCR22, SC7YT and PS12A02, and the nucleotide sequences of the forward primer and the reverse primer are as follows:
(1) PTCR22: the forward primer sequence is shown as SEQ ID NO.29, and the reverse primer sequence is shown as SEQ ID NO. 30;
(2) SC7YT: the forward primer sequence is shown as SEQ ID NO.31, and the reverse primer sequence is shown as SEQ ID NO. 32;
(3) PS12a02: the forward primer sequence is shown as SEQ ID NO.19, and the reverse primer sequence is shown as SEQ ID NO. 20;
the varieties are 42 as follows:
2. use of a kit in sakura variety identification, wherein the kit comprises the sakura SSR molecular marker primer combination according to claim 1, and the sakura variety is 42 varieties listed in claim 1.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434646A (en) * | 2016-11-30 | 2017-02-22 | 宁波城市职业技术学院 | Four pairs of EST-SSR (Expressed Sequence Tag-Simple Sequence Repeat) primers as well as preparation method thereof and application thereof to construction of fingerprint map of cerasus plants |
| CN106957914A (en) * | 2017-04-08 | 2017-07-18 | 中国农业科学院郑州果树研究所 | The authentication method of cherry variety |
| CN111349717A (en) * | 2020-05-06 | 2020-06-30 | 北京市林业果树科学研究院 | SSR (simple sequence repeat) markers of sweet cherry stock resources and fingerprint spectrum database thereof |
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2021
- 2021-10-22 CN CN202111238341.9A patent/CN113981124B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106434646A (en) * | 2016-11-30 | 2017-02-22 | 宁波城市职业技术学院 | Four pairs of EST-SSR (Expressed Sequence Tag-Simple Sequence Repeat) primers as well as preparation method thereof and application thereof to construction of fingerprint map of cerasus plants |
| CN106957914A (en) * | 2017-04-08 | 2017-07-18 | 中国农业科学院郑州果树研究所 | The authentication method of cherry variety |
| CN111349717A (en) * | 2020-05-06 | 2020-06-30 | 北京市林业果树科学研究院 | SSR (simple sequence repeat) markers of sweet cherry stock resources and fingerprint spectrum database thereof |
Non-Patent Citations (2)
| Title |
|---|
| Simple sequence repeat-based assessment of genetic relationships among Prunus rootstocks;Z.Turkoglu等;Genetics and Molecular Research;第9卷(第4期);第4页结果部分、表2 * |
| 樱属植物茎、叶结构及SSR遗传多样性分析;张文昕;中国优秀硕士学位论文全文数据库 农业科技辑;摘要、表4.1 * |
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