CN105112523A - SSR (Simple Sequence Repeats) core primer group developed on basis of whole-genome sequences of cabbages and application - Google Patents

SSR (Simple Sequence Repeats) core primer group developed on basis of whole-genome sequences of cabbages and application Download PDF

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CN105112523A
CN105112523A CN201510530015.3A CN201510530015A CN105112523A CN 105112523 A CN105112523 A CN 105112523A CN 201510530015 A CN201510530015 A CN 201510530015A CN 105112523 A CN105112523 A CN 105112523A
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ssr
chinese cabbage
primer group
application
primers
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CN105112523B (en
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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
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CROP Research Institute of Shandong Academy of Agricultural Sciences
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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
    • 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

Abstract

The invention discloses an SSR (Simple Sequence Repeats) core primer group developed on basis of whole-genome sequences of cabbages and application. The SSR core primer group comprises 17 pairs of primers. Nucleotide sequences of the primers are sequentially shown as sequence tables SEQ ID No.1-34. The SSR core primer group and the application have the advantages that the 17 pairs of screened SSR core primers are uniformly distributed in genomes of the cabbages, banding patterns are clear and are easy to interpret, and the SSR core primer group is suitable for carrying out detection by the aid of common denature polyacrylamide gel electrophoresis detection platforms and capillary fluorescence detection platforms; the SSR core primer group is high in polymorphism and good in amplification repeatability and can be used in the fields of cabbage genetic diversity analysis, variety identification, DNA (deoxyribonucleic acid) fingerprint spectrum construction and the like.

Description

Based on SSR core primers group and the application of the exploitation of Chinese cabbage whole genome sequence
Technical field
The present invention relates to Chinese cabbage SSR marker, specifically based on SSR core primers group and the application of the exploitation of Chinese cabbage whole genome sequence, 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 requisite on dining table especially, therefore has saying of " winter Chinese cabbage beautiful as bamboo shoot ".Chinese cabbage has higher nutritive value, has the saying of " hundred dishes are not as Chinese cabbage ".Containing a large amount of robust fibre in Chinese cabbage, can promote that intestines wall is wriggled, help digest, Chinese cabbage is clearly delicious, Appetizing spleen-tonifying, containing the mineral substance such as protein, fat, multivitamin and calcium, phosphorus, iron, normal food contributes to enhancing body immunologic function, also helpful to defatting beauty.Effective constituent in Chinese cabbage can reduce body's cholesterol level, increase blood vessel elasticity, and normal food can atherosis and some cardiovascular disorder of prevention of arterial.Therefore, can develop, be processed into the food of a series of instant.
Chinese cabbage is that China originates in vegetables, has long cultivation history, accounts for 9% ~ 15% of sowing of vegetable area in the cultivated area of China, and successively imports the countries such as Japan, Korea S, Korea into.Due to Chinese cabbage China and East Asian countries Vegetable produce and consumption in occupy very important status, therefore New Chinese Cabbage Variety cultivate have great importance.In recent years, China's New Chinese Cabbage Variety is cultivated and is presented good growth momentum, cultivates a large amount of new variety.Accurately, carry out the qualification of variety of crops fast, Crop breed audit, kind protection, true and false kind are distinguished, the aspect such as the property right dispute of kind all plays an important role.Traditional cultivar identification method is field trapping test, under identification of species is planted in identical growth conditions, make observation in each stage such as seedling, flowering period, ripening stage to multiple qualitative character, quantitative character and disease resistance etc. to record, and carry out variety protection, the similarities and differences of identification of species.There is long, easy affected by environment, the problem such as test character is many, workload is large qualification cycle in traditional category authentication method, cannot adapt to the qualification requirement of a large amount of kind.
Molecule marker is the genetic marker by between individuality based on nucleotide sequence variation, is the direct reflection of DNA level genetic polymorphism.Markers for Detection technology has restriction in short, not affected by environment and season test period, do not have tissue specificity, selective marker number many, can the advantages such as high-throughput test analysis be carried out, in progressively detecting for cultivar identification, seed purity and variety authentication.Utilize RAPD, AFLP, RFLP to mark, the Chinese cabbage cultivar of different ecological type can be differentiated.But RAPD repeatability is poor; AFLP operation is more complicated, less stable; RFLP operating process is loaded down with trivial details, and efficiency is low, and cost is high.SSR (SimpleSequenceRepeats) is the DNA sequence dna in units of 1 ~ 6 Nucleotide, in tandem sequence repeats, is distributed widely in eukaryotic gene group.Compared with other molecule markers, SSR marker has that codominance, polymorphism are high, reproducible, simple operation and other advantages, is widely used in the fields such as Genetic Diversity research, cultivar identification, the structure of genetic map, QTL location, marker assisted selection and Comparative genomic strategy research as important DNA marker.Compared with the crop such as paddy rice, wheat, existing Chinese cabbage SSR marker quantity is few, can not meet the needs of Chinese cabbage research, and the SSR marker that a large amount of exploitation covers full-length genome is still one of important process of current Chinese cabbage research.
Summary of the invention
The object of the present invention is to provide a set of SSR core primers group based on the exploitation of Chinese cabbage whole genome sequence, these primers have that amplification is stable, electrophoretic band is clear, the advantage of rich polymorphism, can be effective to the researchs such as Chinese cabbage analysis of genetic diversity, cultivar identification, DNA fingerprinting structure.
Technical scheme of the present invention is: based on the SSR core primers group of Chinese cabbage whole genome sequence exploitation, it is characterized in that, it comprises 17 pairs of primers, be 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, CRIB8-12, CRIB9-3 and CRIB9-8, its nucleotide sequence is successively as shown in sequence table SEQ IDNo.1 ~ 34.
The above-mentioned application of SSR core primers group in Chinese cabbage analysis of genetic diversity, cultivar identification, DNA fingerprinting structure.
Beneficial effect of the present invention: 17 pairs of SSR core primers (i.e. SSR marker) that the present invention filters out are evenly distributed in Chinese cabbage genome, the clear easy interpretation of banding pattern, is applicable to common denaturing polyacrylamide gel electrophoresis detection platform and kapillary fluoroscopic examination detection of platform simultaneously; Polymorphism is high; Increase reproducible; can be used for the fields such as Chinese cabbage analysis of genetic diversity, cultivar identification, DNA fingerprinting structure; be conducive to the legitimate rights and interests protecting breeder, producers and consumers, promote the raising of Chinese cabbage genetic breeding level and the development of Chinese cabbage industry.
Accompanying drawing explanation
Fig. 1 is SSRHunter1.3 software search result schematic diagram;
Fig. 2 is the polymorphism that primer detects in Chinese cabbage; Wherein A, B, C figure is respectively the polymorphism that primer CRIB6-23, CRIB5-26 and CRIB9-8 detect in Chinese cabbage, as can be seen from the figure: the polymorphism of primer is the abundantest and banding pattern 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) material 37 parts of Chinese cabbage cultivars (table 1)
Table 137 part Chinese cabbage cultivar
DNA sequence number Variety name DNA sequence number 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 blue or green 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 Dragon red No. 1 of garden
10 Packet header, Fushan 29 The capital autumn 56
11 Western 388-1 30 Dark green king
12 Beautiful blue or green 31 Autumn green 55
13 Z61-2-3 32 Yuzao No.1
14 734-11 33 Green star large dish
15 726-2 34 In selected white 81
16 No. 1, South Road 35 In white 76
17 127-1 36 Section sprouts 75
18 W34-2 37 Autumn green 60
19 W38-1
(2) CTAB method extracts genomic dna
CTAB method is adopted to extract the DNA of experimental cultivar: to get tender tissue compound sample 0.2g, put into 1.5mL centrifuge tube, grind in liquid nitrogen.Or seed is ground, put into 1.5mL centrifuge tube.DNA extraction liquid is preheating to 65 DEG C, and often pipe adds 400 μ L, mixing sample.Centrifuge tube is placed in 65 DEG C of metal baths or water-bath, takes off after insulation 23min, shake centrifuge tube in insulating process, sample is fully mixed with extracting solution.400 μ L24:1 chloroform-isoamyl alcohol (V/V) are added, vibration mixing in centrifuge tube.The centrifugal 10min of 10000g.Supernatant liquor 200 μ L is proceeded to another 1.5mL centrifuge tube, adds 400 μ L-20 DEG C of precooling dehydrated alcohol precipitation DNA.The centrifugal 1min of 10000g, abandons supernatant liquor, adds 500 μ L Ethanol-Acetic Acid ammonium solutions (take 154.6mg ammonium acetate, add 140ml dehydrated alcohol, be settled to 200mL with deionized water), the centrifugal 5min collecting precipitation of 6000g.Room temperature is dried, and adds 100 μ LTE (pH8.0) solubilize DNA, detects DNA concentration.-20 DEG C of preservations.
Wherein, the formula of DNA extraction liquid is: take 20.0g cetyltriethylammonium bromide respectively and 81.82g sodium-chlor puts into beaker, then 40mL disodium EDTA solution (pH8.0), 100mL1mol/L Tutofusin tris hydrochloric acid soln (pH8.0) and 10.0g polyvinylpyrrolidone (PVP) is added, add 800ml deionized water again, heating for dissolving in 65 DEG C of water-baths, is settled to 1000mL after cooling.Sterilizing 20min under 103.4kPa (121 DEG C) condition.In 4 DEG C of preservations.
2. the exploitation of Chinese cabbage genome SSR primer
(1) acquisition of Chinese cabbage genome sequence
Chinese cabbage genome sequencing result is downloaded from Chinese cabbage genome database BrassicaDatabase (http://brassicadb.org/brad/downloadOverview.php), and about 277Mb, comprises 40550 sequences altogether.
(2) qualification of SSR sequence in Chinese cabbage whole genome sequence
In units of karyomit(e), every bar karyomit(e) chooses about 150 sequences (accounting for 45% of every bar chromosome sequence), adopts SSR search software SSRHunter1.3 to carry out the search of SSR site to sequence.Search condition is: multiplicity is at least 5, forms the few nucleotide repeating primitive is at most 6.Statistics perfect form SSR, forme fruste SSR and compound SSR.SSRHunter1.3 software search result as shown in Figure 1.
(3) design of Chinese cabbage genome SSR primer
According to the SSR site that step (2) obtains, select multiplicity more than 10 or repeat the SSR site of base number more than 20, utilizing the conservative flanking sequence at tumor-necrosis factor glycoproteins two ends, design primer with Primer5.0.The condition of design of primers is: PCR primer length is 100 ~ 350bp; Annealing temperature (Tm) is 50 ~ 70 DEG C, ensures that between two primers, Tm value difference is no more than 4 DEG C; GC% content is 40 ~ 65%; Primer length is 18 ~ 28bp; Primer 5 ' holds preferably G/C, and 3 ' end is avoided occurring A.In order to ensure the specificity of primer, for designing conservative flanking sequence and microsatellite locus at least 20 ~ 23, the interval base of primer.Successful design goes out 240 pairs of primers, is synthesized by Jin Weizhi bio tech ltd, Suzhou.
3. the screening of Chinese cabbage genome SSR primer
Select 8 parts of Chinese cabbage cultivars, for detecting amplification situation and the polymorphism of Chinese cabbage genome SSR marker.
Adopt 240 pairs of primers of synthesis, increase with the genomic dna of 8 parts of materials, according to amplification, screening Absorbable organic halogens increases, banding pattern is clear and the primer of rich polymorphism.
Pcr amplification adopts the reaction volume of 25 μ L, and containing often kind of dNTP0.25mmol/L, each 0.4 μm of ol/L of forward primer, reverse primer, Taq DNA polymerase 1.0 unit, 1 × PCR damping fluid is not (containing Mg 2+), MgCl 21.5mmol/L, sample DNA 10-40ng.Response procedures is: 94 DEG C of denaturation 4min; 94 DEG C of sex change 45s, 65 DEG C of annealing 45s, 72 DEG C extend 45s, every cycle down 1 DEG C, totally 15 circulations; 94 DEG C of sex change 45s, 50 DEG C of annealing 30s, 72 DEG C extend 45s, totally 30 circulations; 72 DEG C extend 10min, 4 DEG C of preservations.
According to the selection result, choose the abundantest and banding pattern primer (as shown in Figure 2) clearly of 2 ~ 4 pairs of polymorphisms from Chinese cabbage every bar karyomit(e), totally 17 to (table 2).
Table 217 pair primer
3. utilize 17 pairs of SSR primer pairs, 37 parts of Chinese cabbage cultivars to carry out pcr amplification and capillary electrophoresis
Pcr amplification adopts the reaction volume of 25 μ L, and containing often kind of dNTP0.25mmol/L, each 0.4 μm of ol/L of forward primer, reverse primer, Taq DNA polymerase 1.0 unit, 1 × PCR damping fluid is not (containing Mg 2+), MgCl 21.5mmol/L, sample DNA 10-40ng.Response procedures is: 94 DEG C of denaturation 4min; 94 DEG C of sex change 45s, 65 DEG C of annealing 45s, 72 DEG C extend 45s, every cycle down 1 DEG C, totally 15 circulations; 94 DEG C of sex change 45s, 50 DEG C of annealing 30s, 72 DEG C extend 45s, totally 30 circulations; 72 DEG C extend 10min, 4 DEG C of preservations.
Capillary electrophoresis carries out according to following steps: fluorescently-labeled for 6-FAM and HEX PCR primer ultrapure water is diluted 30 times, and the fluorescently-labeled PCR primer of TAMRA and ROX dilutes 10 times.After getting isopyknic above-mentioned 4 kinds of dilutions respectively, solution mixes, and draws 1 μ L and join in the special deep hole plate hole of DNA analysis instrument from mixed solution.In plate, each hole adds mark and 8.9 μ L deionized formamides in 0.1 μ LLIZ500 molecular weight respectively.By sample 95 DEG C of sex change 5min in PCR instrument, take out, be placed in immediately on trash ice, cooling more than 10min.Be placed to after brief centrifugation 10s (ABI3730XL) on DNA analysis instrument.Except testing sample, each SSR site also should comprise the amplified production of 3 ~ 4 Reference cultivars simultaneously.
Analyze capillary electrophoresis result.According to the amplified fragments size of Reference cultivars, determine the allelic variation size in this site of testing sample.The allelic variation in site of isozygotying is recorded as X/X, and the allelic variation of heterozygous sites is recorded as X/Y, and wherein X, Y are two different allelic variation clip size on this site, and small segment is front, and large fragment is rear.Invalid allelic variation is recorded as 0/0, and the data integration in multiple site forms the SSR finger printing (table 3) of different Chinese cabbage cultivar together.Judge according to the detected result of 17 pairs of SSR primers: Differences number of sites >=3 are as different varieties; Differences number of sites <3 is approximate kind, forms qualification result.Relatively the finger print data of 37 parts of kinds, finds that any two interracial difference number of sites are all greater than 3, illustrates that these 17 pairs of core primers effectively can be opened these Variety identification.
Utilize the number of alleles (Na) of PowerMarkerV3.25 computed in software primer, gene diversity index (He), polymorphism information amount (PIC).Utilize the interracial genetic similarity of NTSYS-pcV2.10e computed in software, carry out cluster analysis by UPGMA method, draw dendrogram (Fig. 3).As can be seen from Figure 3: the genetic similarity index of most of kind is less than 0.05, all kinds can be differentiated.
The finger print data of table 337 part kind
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 group of Chinese cabbage whole genome sequence exploitation, it is characterized in that, it comprises 17 pairs of primers, be 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, CRIB8-12, CRIB9-3 and CRIB9-8, its nucleotide sequence is successively as shown in sequence table SEQ IDNo.1 ~ 34.
2. the application of SSR core primers group according to claim 1 in Chinese cabbage analysis of genetic diversity.
3. the application of SSR core primers group according to claim 1 in Chinese cabbage cultivar qualification.
4. the application of SSR core primers group according to claim 1 in Chinese cabbage DNA fingerprinting structure.
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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
CN108977563A (en) * 2018-08-09 2018-12-11 山东省农业科学院作物研究所 SSR core primers group and its application based on the exploitation of radish whole genome sequence
CN110195126A (en) * 2019-07-16 2019-09-03 山西省农业科学院农作物品种资源研究所 SSR core primers group and its application based on bitter buckwheat full-length genome data mining

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