CN105112523B - SSR core primers group and application based on the exploitation of Chinese cabbage whole genome sequence - Google Patents

SSR core primers group and application based on the exploitation of Chinese cabbage whole genome sequence Download PDF

Info

Publication number
CN105112523B
CN105112523B CN201510530015.3A CN201510530015A CN105112523B CN 105112523 B CN105112523 B CN 105112523B CN 201510530015 A CN201510530015 A CN 201510530015A CN 105112523 B CN105112523 B CN 105112523B
Authority
CN
China
Prior art keywords
chinese cabbage
ssr
primers
ssr core
core primers
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
Application number
CN201510530015.3A
Other languages
Chinese (zh)
Other versions
CN105112523A (en
Inventor
李汝玉
张晗
段丽丽
郑永胜
王雪梅
孙加梅
王东建
李华
仙丽娜
王秀娟
王玮
王穆穆
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
CROP Research Institute of Shandong Academy of Agricultural Sciences
Original Assignee
CROP Research Institute of Shandong Academy of Agricultural Sciences
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by CROP Research Institute of Shandong Academy of Agricultural Sciences filed Critical CROP Research Institute of Shandong Academy of Agricultural Sciences
Priority to CN201510530015.3A priority Critical patent/CN105112523B/en
Publication of CN105112523A publication Critical patent/CN105112523A/en
Application granted granted Critical
Publication of CN105112523B publication Critical patent/CN105112523B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

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
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/156—Polymorphic or mutational markers

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Health & Medical Sciences (AREA)
  • Biotechnology (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Immunology (AREA)
  • Mycology (AREA)
  • Microbiology (AREA)
  • Molecular Biology (AREA)
  • Botany (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)

Abstract

The invention discloses the SSR core primers group developed based on Chinese cabbage whole genome sequence and application.It includes 17 pairs of primers, and its nucleotide sequence is successively as shown in sequence table SEQ ID No.1~34.17 pairs of SSR core primers that the present invention filters out are evenly distributed in Chinese cabbage genome, banding pattern clearly easy interpretation, while are adapted to common denaturing polyacrylamide gel electrophoresis detection platform and capillary fluoroscopic examination detection of platform；Polymorphism is high；Expand it is reproducible, available for Chinese cabbage analysis of genetic diversity, cultivar identification, DNA fingerprinting structure etc. field.

Description

SSR core primers group and application based on the exploitation of Chinese cabbage whole genome sequence
Technical field
The present invention relates to Chinese cabbage SSR marker, specifically the SSR cores based on the exploitation of Chinese cabbage whole genome sequence Primer sets and application, belong to technical field of molecular biology.
Background technology
Chinese cabbage is Cruciferae biennial herb plant, and in the winter of northern China, Chinese cabbage is even more must not on dining table Can be less, therefore have saying for " winter Chinese cabbage being beautiful such as bamboo shoot ".Chinese cabbage has higher nutritive value, there is saying for " hundred dishes being not so good as Chinese cabbage " Method.Containing substantial amounts of crude fibre in Chinese cabbage, intestinal wall can be promoted to wriggle, helped digest, Chinese cabbage is delicious salubrious, Appetizing spleen-tonifying, Containing the mineral matter such as protein, fat, multivitamin and calcium, phosphorus, iron, often feeding helps to strengthen body's immunity, to subtracting It is fertile vigorous and graceful also helpful.Active ingredient in Chinese cabbage can reduce body's cholesterol level, increase blood vessel elasticity, and often feeding can prevent Atherosclerosis and some angiocardiopathies.Therefore, it can develop, be processed into a series of food of instants.
Chinese cabbage is that China originates in vegetables, there is long cultivation history, and sowing of vegetable face is accounted in the cultivated area of China Long-pending 9%~15%, and the successively country such as incoming Japan, South Korea, Korea.Because Chinese cabbage is in the vegetable of China and East Asian countries Occupy very important status in dish production and consumption, therefore New Chinese Cabbage Variety is cultivated and had great importance.In recent years, I State's New Chinese Cabbage Variety, which is cultivated, is presented good growth momentum, cultivates a large amount of new varieties.Accurately, variety of crops is quickly carried out Identification, for Crop breed audit, kind protection, true and false kind distinguish, the property right dispute of kind etc. have it is important Effect.Traditional cultivar identification method is field trapping test, is that identification of species is planted under identical growth conditions, in children Each stage such as seedling, florescence, maturity period makes observation to multiple qualitative characters, quantitative character and disease resistance etc. and recorded, and goes forward side by side Row variety protection, the similarities and differences of identification of species.Traditional category authentication method exist qualification cycle it is long, easily affected by environment, testability The problems such as shape is more, workload is big, the qualification requirement of a large amount of kinds can not be adapted to.
Molecular labeling is the genetic marker based on nucleotide sequence variation between individual, is DNA level genetic polymorphism Direct reflection.Markers for Detection technology has short test period, not affected by environment and season limit, without organizing specific Property, selective marker number is more, can carry out the advantages such as high flux test analysis, it is pure to be progressively used for cultivar identification, seed In degree and variety authentication detection.Marked using RAPD, AFLP, RFLP, can be by the Chinese cabbage cultivar area of different ecological type Do not open.But RAPD repeatability is poor；AFLP operates more complicated, less stable；RFLP operating process is cumbersome, and efficiency is low, into This height.SSR (Simple Sequence Repeats) be in units of 1~6 nucleotides, in the DNA sequence dna of tandem sequence repeats, It is distributed widely in eukaryotic gene group.Compared with other molecular labelings, SSR marker has codominance, polymorphism height, repeated Property good, simple operation and other advantages, as important DNA marker be widely used in Genetic Diversity research, kind mirror The fields such as the fixed, structure of genetic map, QTL positioning, marker assisted selection and Comparative genomic strategy research.Make with rice, wheat etc. Thing is compared, and existing Chinese cabbage SSR marker quantity is few, it is impossible to meets the needs of Chinese cabbage research, a large amount of exploitation covering full genomes The SSR marker of group is still one of important process of current Chinese cabbage research.
The content of the invention
It is an object of the invention to provide it is a set of based on Chinese cabbage whole genome sequence exploitation SSR core primers groups, this A little primers have the advantages of amplification is stable, electrophoretic band is clear, rich polymorphism, can be effectively used for Chinese cabbage genetic diversity point The researchs such as analysis, cultivar identification, DNA fingerprinting structure.
The technical scheme is that：Based on the SSR core primers groups of Chinese cabbage whole genome sequence exploitation, its feature It is that it includes 17 pairs of primers, is respectively：CRIB1-3、CRIB2-4、CRIB2-23、CRIB3-22、CRIB3-28、CRIB4-10、 CRIB4-15、CRIB5-8、CRIB5-11、CRIB5-26、CRIB6-3、CRIB6-23、CRIB7-19、CRIB8-5、CRIB8- 12nd, CRIB9-3 and CRIB9-8, its nucleotide sequence is successively as shown in sequence table SEQ ID No.1~34.
Above-mentioned SSR core primers group is in Chinese cabbage analysis of genetic diversity, cultivar identification, DNA fingerprinting structure side The application in face.
Beneficial effects of the present invention：17 pairs of SSR core primers (i.e. SSR marker) that the present invention filters out are in Chinese cabbage gene It is evenly distributed in group, banding pattern clearly easy interpretation, while is adapted to common denaturing polyacrylamide gel electrophoresis detection platform and hair Tubule fluoroscopic examination detection of platform；Polymorphism is high；Expand it is reproducible, available for Chinese cabbage analysis of genetic diversity, kind reflect The fields such as fixed, DNA fingerprinting structure, are advantageous to protect breeder, the legitimate rights and interests of producers and consumers, promote Chinese cabbage The horizontal raising of genetic breeding and the development of Chinese cabbage industry.
Brief description of the drawings
Fig. 1 is SSRHunter1.3 software search result schematic diagrams；
Fig. 2 is the polymorphism that primer detects in Chinese cabbage；Wherein A, B, C figure are respectively primer CRIB6-23, CRIB5- The polymorphism that 26 and CRIB9-8 is detected in Chinese cabbage, as can be seen from the figure：The polymorphism of primer is most abundant and banding pattern is clear It is clear；
Fig. 3 is the dendrogram of 17 pairs of primer pairs, 37 kinds.
Embodiment
1. the extraction of Chinese cabbage genomic DNA
(1) 37 parts of Chinese cabbage cultivars (table 1) of material
1 37 parts of Chinese cabbage cultivars of table
DNA sequence numbers Variety name DNA sequence numbers Variety name
1 W4-1 20 W33-2
2 W13-8 21 W32-5
3 W28-8 22 141-3
4 659 23 W46-6
5 695-7 24 Beautiful green grass or young crops 2
6 267-1 25 Gold zone baby
7 Jin Bei -2 26 Tianjin green 2
8 229-1 27 Tianjin green 3
9 W3 28 Imperial red No. 1 of garden
10 Fushan packet header 29 The capital autumn 56
11 Western 388-1 30 Dark green king
12 It is beautiful blue or green 31 Autumn green 55
13 Z61-2-3 32 Yuzao No.1
14 734-11 33 Green big dish of star
15 726-2 34 White 81 in selected
16 South Road 1 35 In white 76
17 127-1 36 Section sprouts 75
18 W34-2 37 Autumn green 60
19 W38-1
(2) CTAB methods extraction genomic DNA
Using the DNA of CTAB methods extraction experimental cultivar：Tender tissue aggregate sample 0.2g is taken, is put into 1.5mL centrifuge tubes, Ground in liquid nitrogen.Or grind seed, it is put into 1.5mL centrifuge tubes.DNA extract solutions are preheating to 65 DEG C, often pipe adds 400 μ L, mix sample.Centrifuge tube is placed in 65 DEG C of metal baths or water-bath, be incubated 23min after remove, shaken in insulating process from Heart pipe, makes sample be sufficiently mixed with extract solution.400 μ L 24 are added into centrifuge tube:1 chloroform-isoamyl alcohol (V/V), vibration are mixed It is even.10000g centrifuges 10min.The μ L of supernatant 200 are transferred to another 1.5mL centrifuge tubes, it is anhydrous to add 400 μ L-20 DEG C precoolings Ethanol precipitation DNA.10000g centrifuges 1min, abandons supernatant, adds 500 μ L Ethanol-Acetic Acids ammonium salt solutions and (weighs 154.6mg acetic acid Ammoniums, 140ml absolute ethyl alcohols are added, 200mL is settled to deionized water), 6000g centrifugations 5min collects precipitation.Room temperature is dried, and is added Enter 100 μ L TE (pH 8.0) solution dissolving DNAs, detect DNA concentration.- 20 DEG C of preservations.
Wherein, the formula of DNA extract solutions is：20.0g cetyltriethylammonium bromides and 81.82g chlorinations are weighed respectively Sodium is put into beaker, then adds 40mL disodium EDTAs solution (pH8.0), 100mL 1mol/L trihydroxy methyl ammonia Methylmethane hydrochloric acid solution (pH8.0) and 10.0g polyvinylpyrrolidones (PVP), add 800ml deionized waters, 65 DEG C of water-baths Middle heating for dissolving, 1000mL is settled to after cooling.Sterilize 20min under the conditions of 103.4kPa (121 DEG C).In 4 DEG C of preservations.
2. the exploitation of Chinese cabbage genome SSR primers
(1) acquisition of Chinese cabbage genome sequence
Chinese cabbage genome sequencing result is from Chinese cabbage genome database Brassica Database (http:// Brassicadb.org/brad/downloadOverview.php) download, about 277Mb, altogether comprising 40550 sequences.
(2) in Chinese cabbage whole genome sequence SSR sequences identification
In units of chromosome, every chromosome chooses about 150 sequences (account for every chromosome sequence 45%), adopts The search of SSR sites is carried out to sequence with SSR search softwares SSRHunter1.3.Search condition is：Number of repetition is at least 5, structure Few nucleotide into repetition primitive is preferably at most 6.Count perfect form SSR, forme fruste SSR and compound SSR. SSRHunter1.3 software search results are as shown in Figure 1.
(3) design of Chinese cabbage genome SSR primers
The SSR sites obtained according to step (2), select number of repetition more than 10 or repeat base number more than 20 SSR sites, using the conservative flanking sequence at repetitive sequence both ends, primer is designed with Primer 5.0.The condition of design of primers is： PCR primer length is 100~350bp；Annealing temperature (Tm) is 50~70 DEG C, and Tm values difference is no more than 4 DEG C between ensureing two primers； GC% contents are 40~65%；Primer length is 18~28bp；The end of primer 5 ' is preferably G/C, and 3 ' ends avoid the occurrence of A.In order to protect The specificity of primer is demonstrate,proved, the conservative flanking sequence for designing primer is at least spaced 20~23 bases with microsatellite locus.Into Work(designs 240 pairs of primers, is synthesized by Suzhou Jin Weizhi bio tech ltd.
3. the screening of Chinese cabbage genome SSR primers
8 parts of Chinese cabbage cultivars are selected, for detecting the amplification situation and polymorphism of Chinese cabbage genome SSR marker.
Using 240 pairs of primers of synthesis, expanded with the genomic DNA of 8 parts of materials, according to amplification, screening can Stabilization expands, banding pattern is clear and the primer of rich polymorphism.
PCR amplifications use 25 μ L reaction volume, each containing every kind of dNTP 0.25mmol/L, forward primer, reverse primer 0.4 μm of ol/L, the unit of Taq archaeal dna polymerases 1.0,1 × PCR buffer solutions (are free of Mg2+), MgCl21.5mmol/L, sample DNA10-40ng.Response procedures are：94 DEG C of pre-degeneration 4min；94 DEG C of denaturation 45s, 65 DEG C of annealing 45s, 72 DEG C of extension 45s, are often followed Ring drops 1 DEG C, totally 15 circulations；94 DEG C of denaturation 45s, 50 DEG C of annealing 30s, 72 DEG C of extension 45s, totally 30 circulate；72 DEG C of extensions 10min, 4 DEG C of preservations.
According to the selection result, 2~4 pairs of polymorphisms of selection are most abundant from every chromosome of Chinese cabbage and banding pattern clearly draws Thing (as shown in Figure 2), totally 17 to (table 2).
2 17 pairs of primers of table
3. enter performing PCR amplification and Capillary Electrophoresis using 17 pairs of SSR primer pairs, 37 parts of Chinese cabbage cultivars
PCR amplifications use 25 μ L reaction volume, each containing every kind of dNTP 0.25mmol/L, forward primer, reverse primer 0.4 μm of ol/L, the unit of Taq archaeal dna polymerases 1.0,1 × PCR buffer solutions (are free of Mg2+), MgCl21.5mmol/L, sample DNA10-40ng.Response procedures are：94 DEG C of pre-degeneration 4min；94 DEG C of denaturation 45s, 65 DEG C of annealing 45s, 72 DEG C of extension 45s, are often followed Ring drops 1 DEG C, totally 15 circulations；94 DEG C of denaturation 45s, 50 DEG C of annealing 30s, 72 DEG C of extension 45s, totally 30 circulate；72 DEG C of extensions 10min, 4 DEG C of preservations.
Capillary Electrophoresis follows the steps below：The PCR primer of 6-FAM and HEX fluorescence labelings is diluted with ultra-pure water 30 times, the PCR primer of TAMRA and ROX fluorescence labelings dilutes 10 times.Solution mixes after taking isometric above-mentioned 4 kinds of dilutions respectively, 1 μ L are drawn from mixed liquor to be added in the special deep hole plate hole of DNA analysis instrument.Each hole is separately added into LIZ500 points of 0.1 μ L in plate Son amount internal standard and 8.9 μ L deionized formamides.Sample is denatured 5min for 95 DEG C in PCR instrument, takes out, is immediately placed on trash ice, Cool down more than 10min.It is placed to after brief centrifugation 10s on DNA analysis instrument (ABI3730XL).In addition to testing sample, each SSR Site should also include the amplified production of 3~4 Reference cultivars simultaneously.
Analyze Capillary Electrophoresis result.According to the amplified fragments size of Reference cultivars, determine the testing sample site etc. Position variation size.The allelic variation in homozygous site is recorded as X/X, and the allelic variation of heterozygous sites is recorded as X/Y, and wherein X, Y is Two different allelic variation clip sizes on the site, small fragment is preceding, and large fragment is rear.Invalid allelic variation is recorded as 0/ 0, the Data Integration in multiple sites forms the SSR finger-prints (table 3) of different Chinese cabbage cultivars together.Draw according to 17 couples of SSR The testing result of thing is judged：Differences number of sites >=3 are different cultivars；Differences number of sites<3 be approximate product Kind, form qualification result.Compare the finger print data of 37 parts of kinds, it is found that any two interracial difference number of sites are both greater than 3, say This bright 17 pairs of core primers effectively can open these Variety identifications.
Using PowerMarker V3.25 softwares calculate the number of alleles (Na) of primer, gene diversity index (He), Polymorphism information amount (PIC).Interracial genetic similarity is calculated using NTSYS-pc V2.10e softwares, is entered with UPGMA methods Row cluster analysis, draw dendrogram (Fig. 3).As can be seen from Figure 3：The genetic similarity index of most of kinds is less than 0.05, all kinds can be differentiated.
The finger print data of 3 37 parts of kinds of table
DNA B7-19 B8-12 B8-5 B1-3 B2-4 B2-23 B5-11 B5-26
1 224/224 227/227 0/0 118/118 180/180 207/207 198/198 280/280
2 218/218 227/227 345/354 118/118 196/196 201/201 198/198 280/280
3 221/221 227/227 357/357 118/118 183/183 207/207 192/192 277/277
4 221/221 227/227 351/351 118/118 183/183 207/207 198/198 277/277
5 218/221 227/233 348/351 112/118 189/189 207/210 154/195 246/280
6 199/199 227/227 351/351 112/112 189/189 207/207 192/192 0/0
7 224/224 227/233 357/357 118/118 189/189 204/204 154/195 277/280
8 224/224 227/227 351/351 118/118 189/189 207/207 192/192 277/277
9 221/224 0/0 351/351 112/118 183/183 207/207 154/192 260/277
10 221/221 233/233 351/351 118/118 189/189 207/207 157/157 277/277
11 221/221 227/227 0/0 118/118 183/183 207/207 198/198 277/277
12 221/221 233/233 357/357 0/0 180/180 0/0 0/0 246/246
13 218/218 233/233 348/348 118/118 0/0 204/204 195/195 260/260
14 221/221 233/233 345/345 118/118 180/180 210/210 154/154 246/246
15 221/221 233/233 351/351 112/112 180/180 204/204 160/160 277/277
16 218/224 227/233 351/351 118/118 0/0 207/207 154/154 277/280
17 221/221 227/227 348/348 118/118 189/189 207/207 192/192 277/277
18 221/221 233/242 345/348 118/118 189/189 0/0 195/195 277/277
19 224/224 227/233 348/348 112/118 189/189 201/207 192/198 246/277
20 218/218 233/233 351/351 112/112 189/189 207/207 195/195 0/0
21 221/221 233/233 351/351 118/118 189/189 207/207 195/195 246/246
22 221/221 242/242 348/354 118/118 183/183 207/207 195/195 260/260
23 218/224 227/242 0/0 106/118 180/180 207/207 195/195 246/246
24 221/221 0/0 0/0 118/118 0/0 210/210 154/154 0/0
25 224/224 233/233 348/348 118/118 0/0 204/207 0/0 277/277
26 221/221 233/233 345/348 112/112 0/0 207/207 192/192 280/280
27 221/221 233/233 351/351 118/118 180/180 210/210 154/154 246/246
28 224/224 227/242 345/348 118/118 180/180 207/207 192/192 277/277
29 221/224 233/242 348/348 118/118 183/183 204/207 157/189 260/283
30 218/221 227/242 0/0 118/118 183/189 204/207 160/160 246/260
31 221/221 233/233 348/348 112/112 180/183 201/201 195/195 246/260
32 218/224 227/233 351/357 112/118 180/183 204/207 185/185 277/283
33 221/221 227/227 351/351 118/118 180/183 207/210 154/204 260/277
34 224/227 227/233 0/0 118/118 180/196 201/207 154/154 269/277
35 221/221 233/242 0/0 118/118 180/180 207/209 195/198 260/277
36 221/221 233/233 0/0 112/118 180/180 207/210 0/0 260/260
37 221/224 233/233 345/348 112/112 180/180 207/207 189/189 260/280
Continued 3
DNA B9-8 B3-22 B3-28 B5-8 B6-3 B6-23 B9-3 B4-10 B4-5
1 191/191 192/192 369/369 197/197 0/0 244/244 0/0 0/0 266/266
2 177/177 186/186 343/343 197/197 210/210 232/232 251/251 245/245 266/266
3 191/191 183/183 391/391 197/197 216/216 262/262 255/255 245/245 256/256
4 177/177 189/189 331/331 197/197 216/216 244/244 246/246 0/0 266/266
5 191/191 186/186 0/0 193/193 210/210 244/262 255/255 225/225 266/266
6 191/191 180/180 331/331 197/197 213/213 262/262 216/216 216/245 266/266
7 179/179 189/189 0/0 193/193 210/210 244/262 251/251 216/216 266/266
8 188/188 189/189 371/371 197/197 216/216 262/262 232/232 245/245 275/275
9 191/197 183/183 360/371 177/177 210/210 250/262 251/251 245/245 260/260
10 179/179 189/189 374/374 177/177 210/210 244/244 250/250 216/216 272/272
11 177/177 189/189 331/331 197/197 216/216 244/244 246/246 245/245 266/266
12 179/191 189/189 371/371 0/0 0/0 244/244 255/255 0/0 256/256
13 191/191 189/189 366/366 193/193 210/210 244/244 250/250 216/225 266/266
14 179/179 189/189 371/371 193/193 0/0 244/244 259/259 225/225 256/256
15 177/177 189/189 331/360 195/195 204/204 244/244 280/280 216/216 260/260
16 179/191 195/195 360/360 193/203 176/210 256/262 246/251 216/225 260/260
17 191/191 183/183 371/371 193/193 213/213 262/262 251/251 245/245 266/266
18 191/191 186/186 372/372 197/197 176/176 0/0 255/255 225/225 266/266
19 179/188 183/195 354/360 197/197 207/210 244/244 246/251 225/225 266/266
20 191/191 192/192 0/0 197/197 210/210 244/244 251/251 225/225 266/266
21 177/177 192/192 371/371 197/197 216/216 250/250 250/250 225/225 266/266
22 179/179 189/189 371/371 190/203 213/213 244/244 246/246 245/245 266/266
23 191/197 189/195 360/377 190/203 207/213 244/250 251/251 216/225 260/266
24 0/0 189/189 0/0 193/203 0/0 244/244 259/259 216/225 266/266
25 191/191 189/189 340/340 203/203 210/210 244/244 250/250 222/222 272/272
26 177/177 192/192 337/337 197/197 204/204 232/232 236/236 213/213 256/256
27 191/191 189/189 374/374 193/193 0/0 244/244 255/255 225/225 256/256
28 177/177 189/189 343/360 203/203 176/213 244/244 255/255 225/225 266/272
29 179/191 183/189 371/371 197/197 207/207 244/244 246/246 225/225 260/275
30 177/188 183/189 346/346 197/197 204/216 244/250 246/255 213/225 266/266
31 177/191 183/192 340/340 203/203 176/204 232/250 251/259 213/216 0/0
32 0/0 183/186 369/372 190/203 176/207 244/244 251/251 225/225 260/260
33 179/188 183/189 369/371 193/193 213/216 244/250 251/255 225/225 260/260
34 179/191 189/195 343/343 203/203 176/210 250/256 251/255 216/225 260/260
35 191/191 192/195 340/385 203/203 176/204 232/250 255/255 245/248 266/272
36 179/179 186/189 0/0 193/193 182/182 244/244 259/259 225/225 256/256
37 177/191 186/192 340/357 203/203 182/204 232/250 236/246 213/216 266/266

Claims (4)

1. based on the SSR core primers groups of Chinese cabbage whole genome sequence exploitation, it is characterized in that, it includes 17 pairs of primers, respectively For：CRIB1-3、CRIB2-4、CRIB2-23、CRIB3-22、CRIB3-28、CRIB4-10、CRIB4-15、CRIB5-8、 CRIB5-11, CRIB5-26, CRIB6-3, CRIB6-23, CRIB7-19, CRIB8-5, CRIB8-12, CRIB9-3 and CRIB9- 8, its nucleotide sequence is successively as shown in sequence table SEQ ID No.1~34.
2. application of the SSR core primers group in terms of Chinese cabbage analysis of genetic diversity described in claim 1.
3. application of the SSR core primers group in terms of Chinese cabbage cultivar identification described in claim 1.
4. application of the SSR core primers group in terms of Chinese cabbage DNA fingerprinting structure described in claim 1.
CN201510530015.3A 2015-08-26 2015-08-26 SSR core primers group and application based on the exploitation of Chinese cabbage whole genome sequence Active CN105112523B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201510530015.3A CN105112523B (en) 2015-08-26 2015-08-26 SSR core primers group and application based on the exploitation of Chinese cabbage whole genome sequence

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201510530015.3A CN105112523B (en) 2015-08-26 2015-08-26 SSR core primers group and application based on the exploitation of Chinese cabbage whole genome sequence

Publications (2)

Publication Number Publication Date
CN105112523A CN105112523A (en) 2015-12-02
CN105112523B true CN105112523B (en) 2017-11-24

Family

ID=54660625

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201510530015.3A Active CN105112523B (en) 2015-08-26 2015-08-26 SSR core primers group and application based on the exploitation of Chinese cabbage whole genome sequence

Country Status (1)

Country Link
CN (1) CN105112523B (en)

Families Citing this family (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108504771A (en) * 2018-06-22 2018-09-07 福建农林大学 A method of exploitation sugarcane SSR marker and identification Sugarcane Breeding affiliation
CN108977563B (en) * 2018-08-09 2019-12-06 山东省农业科学院作物研究所 SSR core primer group developed based on radish whole genome sequence and application thereof
CN110195126B (en) * 2019-07-16 2023-03-21 山西省农业科学院农作物品种资源研究所 SSR core primer group developed based on tartary buckwheat whole genome data and application thereof

Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102220315A (en) * 2011-04-15 2011-10-19 北京市农林科学院 Watermelon complete genomic sequence information based analyzed and developed SSR core primer combinations and application thereof
CN102703586A (en) * 2012-05-18 2012-10-03 北京林业大学 Method for constructing Prunus mume Sieb.et Zucc SSR (simple sequence repeat) genetic map
CN103060318A (en) * 2013-01-11 2013-04-24 山东省农业科学院作物研究所 SSR (Simple Sequence Repeat) core primer group developed based on whole genome sequence of foxtail millet and application of SSR core primer group
CN103667480A (en) * 2013-12-06 2014-03-26 中国农业科学院油料作物研究所 SSR core primer group developed based on sesame complete genomic sequence and application

Patent Citations (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102220315A (en) * 2011-04-15 2011-10-19 北京市农林科学院 Watermelon complete genomic sequence information based analyzed and developed SSR core primer combinations and application thereof
CN102703586A (en) * 2012-05-18 2012-10-03 北京林业大学 Method for constructing Prunus mume Sieb.et Zucc SSR (simple sequence repeat) genetic map
CN103060318A (en) * 2013-01-11 2013-04-24 山东省农业科学院作物研究所 SSR (Simple Sequence Repeat) core primer group developed based on whole genome sequence of foxtail millet and application of SSR core primer group
CN103667480A (en) * 2013-12-06 2014-03-26 中国农业科学院油料作物研究所 SSR core primer group developed based on sesame complete genomic sequence and application

Non-Patent Citations (4)

* Cited by examiner, † Cited by third party
Title
大白菜品种鉴定的SSR核心引物筛选及其应用;隋光磊;《园艺学报》;20141031;第41卷(第10期);第2021-2034页 *
大白菜简单序列重复(SSR)和***/缺失(InDel)标记的开发及通用性;仪泽会;《农业生物技术学报》;20121225;第20卷(第12期);第1398-1406页 *
大白菜高通量SSR标记鉴定体系的建立和应用;张晗;《植物遗传资源学报》;20140420;第15卷(第4期);第815-823页 *
植物SSR引物开发策略简述;陈怀琼;《分子植物育种》;20090728;第7卷(第4期);第845-851页 *

Also Published As

Publication number Publication date
CN105112523A (en) 2015-12-02

Similar Documents

Publication Publication Date Title
CN112280881B (en) SNP (Single nucleotide polymorphism) marker combination for identifying broccoli germplasm resources and varieties and application
CN112980999B (en) SSR molecular marker of mulberry variety Yuehei 74 and core primer group, kit and application thereof
CN103194537A (en) Cabbage SSR fingerprint construction method
CN111961750A (en) KASP primer for detecting tomato yellow leaf curl virus disease resistance gene Ty-1 and application thereof
CN105112523B (en) SSR core primers group and application based on the exploitation of Chinese cabbage whole genome sequence
CN110331217B (en) Microsatellite marker paternity test primer suitable for Nile tilapia, Oreochromis aureus and hybrid thereof, method and application
CN114262748B (en) Molecular marker for identifying variety &#39;Yueshi 143&#39;, identifying primer group, kit and application
CN111961749A (en) KASP primer for detecting tomato yellow leaf curl virus disease resistance genes Ty-3 and Ty-3a and application thereof
KR20090035303A (en) Multiplex real time pcr method for discrimination of rice cultivar
CN112602589B (en) Method for rapid and accurate breeding of semi-waxy japonica rice restorer
CN108977563B (en) SSR core primer group developed based on radish whole genome sequence and application thereof
CN116179748B (en) Molecular marker primer group and kit for identifying fruit Sang Pinchong &#39;Yue mulberry 33&#39; and application thereof
KR102192664B1 (en) Identification of kiwifruit using SCAR markers and multiplex PCR assays
CN116926229A (en) Molecular marker closely linked with high-vitamin E main effect QTL locus qVE.C02 of rape seed and application
CN110878376A (en) SSR molecular marker primer for identifying dendrobium huoshanense and application thereof
CN107385052B (en) STR primer for identifying clone of eucalyptus and application thereof
CN114107551B (en) Molecular marker for identifying or assisting in identifying soybean protein content and application thereof
CN113832251B (en) SNP locus combination for detecting tomato mosaic virus resistance and application thereof
CN113637791B (en) Molecular marker for simultaneously identifying restorability and authenticity of pepper male sterile three-line hybrid and identification method thereof
CN107338246B (en) Specific sequence of tomato fruit dry juice character and its molecular marker and identification method
WO2023070937A1 (en) Ssr marker for detecting bruchus rufimanus boheman-resistant variety of vicia faba l. and use thereof
CN112575101B (en) Molecular marker related to resistance of Cucurbita pepo PRSV-W virus disease and application thereof
KR20160057021A (en) HRM Primer sets for discriminating Korean and American ginseng and uses thereof
CN116334269B (en) Molecular marker for sex identification of actinidia arguta and specific primer pair M3 thereof
CN113736906B (en) SNP locus combination for detecting verticillium wilt resistance of tomatoes and application thereof

Legal Events

Date Code Title Description
C06 Publication
PB01 Publication
C10 Entry into substantive examination
SE01 Entry into force of request for substantive examination
GR01 Patent grant
GR01 Patent grant