CN105543386A - Molecular marker tightly interlocked with soybean seed coat color and application of molecular marker - Google Patents
Molecular marker tightly interlocked with soybean seed coat color and application of molecular marker Download PDFInfo
- Publication number
- CN105543386A CN105543386A CN201610069040.0A CN201610069040A CN105543386A CN 105543386 A CN105543386 A CN 105543386A CN 201610069040 A CN201610069040 A CN 201610069040A CN 105543386 A CN105543386 A CN 105543386A
- Authority
- CN
- China
- Prior art keywords
- primer pair
- pcr amplification
- correspondence
- amplification product
- product size
- 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.)
- Granted
Links
- 244000068988 Glycine max Species 0.000 title claims abstract description 97
- 235000010469 Glycine max Nutrition 0.000 title claims abstract description 94
- 239000003147 molecular marker Substances 0.000 title abstract description 11
- 238000012408 PCR amplification Methods 0.000 claims description 143
- 238000001514 detection method Methods 0.000 claims description 30
- 239000012807 PCR reagent Substances 0.000 claims description 18
- 238000000034 method Methods 0.000 claims description 14
- 238000012360 testing method Methods 0.000 claims description 7
- 238000000137 annealing Methods 0.000 claims description 4
- 238000012797 qualification Methods 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims 1
- 108090000623 proteins and genes Proteins 0.000 abstract description 15
- 238000000926 separation method Methods 0.000 abstract description 5
- 238000009395 breeding Methods 0.000 abstract description 3
- 230000001488 breeding effect Effects 0.000 abstract description 3
- 238000002474 experimental method Methods 0.000 abstract description 3
- 238000010367 cloning Methods 0.000 abstract description 2
- 102000053602 DNA Human genes 0.000 abstract 9
- 108020004414 DNA Proteins 0.000 abstract 9
- 239000003550 marker Substances 0.000 description 34
- 239000000463 material Substances 0.000 description 17
- 239000012896 selective serotonin reuptake inhibitor Substances 0.000 description 12
- 229940124834 selective serotonin reuptake inhibitor Drugs 0.000 description 12
- 241000196324 Embryophyta Species 0.000 description 9
- 238000013507 mapping Methods 0.000 description 8
- 238000011144 upstream manufacturing Methods 0.000 description 8
- 239000003153 chemical reaction reagent Substances 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 238000004458 analytical method Methods 0.000 description 5
- 238000012098 association analyses Methods 0.000 description 5
- 238000001962 electrophoresis Methods 0.000 description 5
- 239000010985 leather Substances 0.000 description 5
- 229930014669 anthocyanidin Natural products 0.000 description 4
- 150000001452 anthocyanidin derivatives Chemical class 0.000 description 4
- 235000008758 anthocyanidins Nutrition 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 229920002770 condensed tannin Polymers 0.000 description 4
- 239000005357 flat glass Substances 0.000 description 4
- 235000018192 pine bark supplement Nutrition 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
- 230000008021 deposition Effects 0.000 description 3
- 239000000975 dye Substances 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 2
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 230000004087 circulation Effects 0.000 description 2
- 230000001276 controlling effect Effects 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 108010060641 flavanone synthetase Proteins 0.000 description 2
- 229930003935 flavonoid Natural products 0.000 description 2
- 150000002215 flavonoids Chemical class 0.000 description 2
- 235000017173 flavonoids Nutrition 0.000 description 2
- 230000007614 genetic variation Effects 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 239000000049 pigment Substances 0.000 description 2
- 229920002401 polyacrylamide Polymers 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 102000004169 proteins and genes Human genes 0.000 description 2
- 230000004044 response Effects 0.000 description 2
- 150000003839 salts Chemical class 0.000 description 2
- SQGYOTSLMSWVJD-UHFFFAOYSA-N silver(1+) nitrate Chemical compound [Ag+].[O-]N(=O)=O SQGYOTSLMSWVJD-UHFFFAOYSA-N 0.000 description 2
- LCGLNKUTAGEVQW-UHFFFAOYSA-N Dimethyl ether Chemical compound COC LCGLNKUTAGEVQW-UHFFFAOYSA-N 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000231392 Gymnosiphon Species 0.000 description 1
- 208000035199 Tetraploidy Diseases 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 229960000583 acetic acid Drugs 0.000 description 1
- 229960000935 dehydrated alcohol Drugs 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 238000012214 genetic breeding Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000012362 glacial acetic acid Substances 0.000 description 1
- 235000021374 legumes Nutrition 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000000877 morphologic effect Effects 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000008844 regulatory mechanism Effects 0.000 description 1
- 238000005204 segregation Methods 0.000 description 1
- 229910001961 silver nitrate Inorganic materials 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/13—Plant traits
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
Abstract
The invention discloses a molecular marker tightly interlocked with a soybean seed coat color and application of the molecular marker. The molecular marker disclosed by the invention is composed of a primer pair 1, a primer pair 2, a primer pair 3 and a primer pair 4, wherein the primer pair 1 is composed of a single-chain DNA (Deoxyribonucleic Acid) molecule represented as a sequence 1 and a single-chain DNA molecule represented as a sequence 2; the primer pair 2 is composed of a single-chain DNA molecule represented as a sequence 3 and a single-chain DNA molecule represented as a sequence 4; the primer pair 3 is composed of a single-chain DNA molecule represented as a sequence 5 and a single-chain DNA molecule represented as a sequence 6; the primer pair 4 is composed of a single-chain DNA molecule represented as a sequence 7 and a single-chain DNA molecule represented as a sequence 8. An experiment shows that the molecular marker disclosed by the invention can be used for effectively identifying the soybean seed coat color; on one hand, fine positioning, separation and cloning of genes can be facilitated; on the other hand, the molecular marker has an important application value for soybean germplasm resource gene type identification and molecular breeding.
Description
Technical field
The present invention relates to a kind of and the closely linked molecule marker of soybean seed coat color and application thereof, belong to biological technical field.
Background technology
Soybean [Glycinemax (L.) Merr.] is the legume crop of extensively plantation in the world, contains rich in protein, containing eight kinds of indispensable amino acids that human body self can not synthesize, extremely important to human nutrition.Soybean seeds oil content and protein content, respectively 20% and about 40%, are the important sources of vegetation fat and feedstuff protein.In Soybean Germplasm classification and genetic breeding, kernel seed coat colour always as the important evidence of soybean resource classification, and is the important morphological mark of breeding progeny assessment.The cultivated soybean evolves from the wild soybean of black race's skin to develop, and has Huang, the kernel seed coat colour enriched such as green, black, brown, two.Soybean Species color of the leather and soybean seeds outward appearance and commercial quality in close relations, it causes the natural product of kernel seed coat colour difference to comprise flavonoid and anthocyanidin, not only have influence on pharmaceutical use and the nutritive value of oxidation-resistance, also in anti-low temperature stress or minimizing virus disease, there is certain effect.Therefore, the research of Soybean Species color of the leather not only has important theory significance, and has practical value.
The genetic research of Soybean Species color of the leather started from for 20 beginnings of the century, had found so far to have at least the participation of 6 sites (I, T, W1, R, O, G) to control the formation (Yangetal.2010 of proterties; Wangetal.2013), exist between site simultaneously and do mutually, hereditary basis complexity (Gaietal.2006).These 3 sites of I, R, T are mainly through controlling the color (Bernardetal.1973 of the synthesis regulation seed coat of pigment; Palmeretal.1987; Nicholasetal.1993).I site controls spatial distribution position and the deposition of anthocyanidin and pycnogenols, suppresses the pigment deposition of entire seed coat, thus causes ripe seed coat to show as uniform yellow.I site is positioned in the region (Toddetal.1996 that soybean No. 8 karyomit(e) one is rich in chalcone synthase (CHS); Cloughetal.2004; Tutejaetal.2004).R and T two sites are by controlling the concrete color (Zabalaetal.2003 of kind controls local of pycnogenols and anthocyanidin; Todaetal.2002), black (i, R, T), imperfect black (i, R, t), brown (i, r, T) and shallow brown (i, r, t) (Yangetal.2010) is comprised.In Black seed coat (i, R, T) and incomplete Black seed coat (i, R, t), anthocyanidin and pycnogenols all have deposition (Buzzetletal.1987; Toddetal.1993).Pycnogenols is only had to synthesize (Zabalaetal.2003) in brown (i, r, T), shallow brown (i, r, t) seed coat.All the other two site O and W1 just just affect kernel seed coat colour (Bernardetal.1973) separately in ir and the it genotype of homozygous recessive, and their may coded flavonoid pathways metabolism relevant enzyme and transhipment enzyme or relevant regulatory factor (Yangetal.2010; Zabalaetal.2007; Xieetal.2003).
Along with appearance and the development of molecular marking technique, in various plants, QTL (Huang, the etal.2012 relevant to economical characters such as output, quality, resistances are located by association analysis and linkage analysis; Liuetal.2007; Qietal.2014; Parketal.2012; Nemrietal.2010).A lot of work has been carried out both at home and abroad in the soybean seed coat color assignment of genes gene mapping and linked marker screening, except the site of classical genetics location, people utilize segregating population in 5 linkage groups, located the relevant QTL (Githirietal.2007 of 8 soybean seed coat color; Oyooetal.2010; Ohnishietal.2011), but due to marker number less, the interval of location is general larger.In these QTL, there are two QTL and I sites closer, but due to comparatively large between positioning area, are difficult to determine that they are exactly a QTL.Also some reports are had the mark in multiple site to be combined the efficiency of selection (GuoandQiu2013 that can increase soybean seed coat color in recent years; Lietal.2014a).
Above-mentioned majority research shows, soybean seed coat color is by the complicated shape of controlled by multiple genes, mutual regulation relationship is there is between different genes, simultaneously, soybean is as the ancient tetraploid plant of one, genome tumor-necrosis factor glycoproteins is more, and Genes location colony is general less, and these factors both increase soybean seed coat color gene Fine Mapping and the difficulty be separated.
Summary of the invention
An object of the present invention is to provide the primer of a kind of detection or auxiliary detection soybean seed coat color proterties to be measured.
The primer of detection provided by the invention or auxiliary detection soybean seed coat color to be measured proterties is following 1)-4):
1) primer set be made up of primer pair 1, primer pair 2, primer pair 3 and primer pair 4:
2) primer pair 1;
3) primer set be made up of primer pair 1 and primer pair 2;
4) primer set be made up of primer pair 1, primer pair 3 and primer pair 4;
Described primer pair 1 is for being made up of the single strand dna shown in the single strand dna shown in sequence 1 and sequence 2;
Described primer pair 2 is for being made up of the single strand dna shown in the single strand dna shown in sequence 3 and sequence 4;
Described primer pair 3 is for being made up of the single strand dna shown in the single strand dna shown in sequence 5 and sequence 6;
Described primer pair 4 is for being made up of the single strand dna shown in the single strand dna shown in sequence 7 and sequence 8.
Another object of the present invention is to provide the PCR reagent of a kind of detection or auxiliary detection soybean seed coat color proterties to be measured.
The PCR reagent of detection provided by the invention or auxiliary detection soybean seed coat color to be measured proterties comprises above-mentioned primer.
In above-mentioned PCR reagent, described primer pair 1, described primer pair 2, described primer pair 3 and the final concentration of described primer pair 4 in described PCR reagent are 0.2 μm of ol/L.
A further object of the invention is to provide the test kit of a kind of detection or auxiliary detection soybean seed coat color proterties to be measured.
The test kit of detection provided by the invention or auxiliary detection soybean seed coat color to be measured proterties comprises above-mentioned primer or above-mentioned PCR reagent.
A further object of the invention is to provide the novelty teabag of above-mentioned primer or above-mentioned PCR reagent or mentioned reagent box.
The invention provides above-mentioned primer or above-mentioned PCR reagent or mentioned reagent box in the application detected or in auxiliary detection soybean seed coat color proterties to be measured.
Present invention also offers the application in the product preparing detection or auxiliary detection soybean seed coat color proterties to be measured of above-mentioned primer or above-mentioned PCR reagent or mentioned reagent box.
In above-mentioned primer or above-mentioned PCR reagent or mentioned reagent box or above-mentioned application, described soybean seed coat color proterties is green, yellow, black or brown.
Last object of the present invention is to provide a kind of method of detection or auxiliary detection soybean seed coat color proterties to be measured.
The method of detection provided by the invention or auxiliary detection soybean seed coat color proterties to be measured is following (A)-(D):
(A) comprise the steps:
A1) with above-mentioned primer pair 1, pcr amplification is carried out to the genomic dna of soybean to be measured, obtain pcr amplification product;
A2) size of described pcr amplification product is detected,
If the pcr amplification product size of primer pair 1 correspondence is 224bp, then soybean seed coat color to be measured be or candidate for green or yellow;
If the pcr amplification product size of primer pair 1 correspondence is 186bp, then soybean seed coat color to be measured is or candidate is black or brown;
B) comprise the steps:
B1) with above-mentioned primer pair 1 and primer pair 2, pcr amplification is carried out to the genomic dna of soybean to be measured respectively, obtain 2 kinds of pcr amplification products;
B2) size of described 2 kinds of pcr amplification products is detected,
If the pcr amplification product size of primer pair 1 correspondence is 224bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp, then soybean seed coat color to be measured be or candidate for green;
If the pcr amplification product size of primer pair 1 correspondence is 224bp, and the pcr amplification product size of primer pair 2 correspondence is 270bp, then soybean seed coat color to be measured be or candidate for yellow;
C) comprise the steps:
C1) with above-mentioned primer pair 1, primer pair 3 and primer pair 4, pcr amplification is carried out to the genomic dna of soybean to be measured respectively, obtain 3 kinds of pcr amplification products;
C2) size of described 3 kinds of pcr amplification products is detected,
If the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 3 correspondence is 288bp, and the pcr amplification product size of primer pair 4 correspondence is 300bp, then soybean seed coat color to be measured is or candidate is black;
If the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp, and the pcr amplification product size of primer pair 4 correspondence is 300bp,
Or the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp or 288bp, and the pcr amplification product size of primer pair 4 correspondence is 294bp, then soybean seed coat color to be measured is or candidate is brown;
D) comprise the steps:
D1) with above-mentioned primer pair 1, primer pair 2, primer pair 3 and primer pair 4, pcr amplification is carried out to the genomic dna of soybean to be measured respectively, obtain 4 kinds of pcr amplification products;
D2) size of described 4 kinds of pcr amplification products is detected,
If the pcr amplification product size of primer pair 1 correspondence is 224bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp or 288bp, and the pcr amplification product size of primer pair 4 correspondence is 294bp or 300bp, then soybean seed coat color to be measured be or candidate for green;
If the pcr amplification product size of primer pair 1 correspondence is 224bp, and the pcr amplification product size of primer pair 2 correspondence is 270bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp or 288bp, and the pcr amplification product size of primer pair 4 correspondence is 294bp or 300bp, then soybean seed coat color to be measured be or candidate for yellow;
If the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp or 270bp, and the pcr amplification product size of primer pair 3 correspondence is 288bp, and the pcr amplification product size of primer pair 4 correspondence is 300bp, then soybean seed coat color to be measured is or candidate is black;
If the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp or 270bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp, and the pcr amplification product size of primer pair 4 correspondence is 300bp; Or the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp or 270bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp or 288bp, and the pcr amplification product size of primer pair 4 correspondence is 294bp, then soybean seed coat color to be measured is or candidate is brown.
In aforesaid method, the annealing temperature of described pcr amplification is 47 DEG C.
Above-mentioned primer or above-mentioned PCR reagent or mentioned reagent box or the application of aforesaid method in Soybean Germplasm qualification and/or soybean breeder also belong to protection scope of the present invention.
Present invention finds and control the site of soybean seed coat color, and develop with these site close linkages, be divided into from molecule marker.Prove by experiment: molecule marker of the present invention effectively can identify soybean seed coat color, contribute to the Fine Mapping of gene on the one hand, be separated and clone, for Soybean Germplasm genotype identification and molecular breeding, all there is important using value on the other hand.
Accompanying drawing explanation
Fig. 1 is the whole-genome association of 56 parts of sequence material seed coat colour of resurveying.
Fig. 2 is the remaining heterozygosis system of product 95-5383 and the hereditary segregating population of salt tolerant 279 in utilizing, and builds the schematic diagram of single-gene segregating population.
Fig. 3 is the Fine Mapping in G site.
Fig. 4 is the Fine Mapping in I site.
Fig. 5 is the Fine Mapping in R site.
Fig. 6 is the regulatory mechanism in 4 sites in F2 colony.
Embodiment
The experimental technique used in following embodiment if no special instructions, is ordinary method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
Embodiment 1, with the acquisition of the closely linked molecule marker of kernel seed coat colour
One, by SNP site that association analysis acquisition is relevant to soybean seed coat color
Do the data that association analysis is used in the present embodiment come from ncbi database (http://www.ncbi.nlm.nih.gov/SNP/snp_viewTable.cgi? handle=NFCRI_MOA_CAAS), by to 56 parts comprise cultivation, place, all SNP of wild soybean storeroom screen, (first group is 176065 SNPs on predicted gene to screen three groups of SNPs altogether, second group is that above-mentioned SNPs removes remaining 98244 SNPs after same sense mutation, and the 3rd group is 94261 SNPs of nonsynonymous mutation).The SNP on gene is positioned at from 20 karyomit(e)s, random choose goes out 299 SNP, for population genetic variations analysis, by Sturcture4.0, the population genetic variations of 56 parts of soybean materials is analyzed, and estimate K value by design factor Δ K, during discovery K=2, Δ K is larger, this colony is divided into and mainly comprises cultivation and two wild subgroups, the method of the mixed linear model analysis of Tassel software and M+K is utilized to carry out whole-genome association, in conjunction with QQ-Plot and practical situation, threshold value is located P<0.001, by the distribution (Fig. 1) of Manhattan figure and institute markd physical location observation mark of correlation, obtain eventually through association analysis and be positioned at 14 interval 146 SNP site relevant to the kernel seed coat colour of soybean (table 1), these sites comprise 5 sites of classical genetics.
Table 1, seed coat colour whole-genome association are interval
Two, the structure of linkage analysis colony and remaining heterozygosis system
With product 95-5383 in the soybean material of yellow seed coat for female parent, the soybean material salt tolerant 279 of brown seed coat color is hybridized for male parent, obtain F2 colony after first familiar generation simple grain passes, each for F2 individual plant kind is become a strain, in F4, F5, F6 each generation, each strain selects a single-strain planting procreation.In F5, F6, identify that seed coat colour segregation ratio meets the individual plant (Fig. 2) of the single-gene regulation and control of 3:1.Meanwhile, utilize F2 and RIL colony to identify related locus, select the individual plant exchanging restructuring to carry out the procreation of expansion colony.
Three, BSA method validate association site of analysis is utilized
According to the type that individual plant seed coat colour is separated, the material mixing be separated by different seed coat colour together, build the gene pool of different seed coat colour, carry out screening (table 2) with the gene pool of SSR primer pair two parents and different seed coat colour of being distributed in association analysis location proximate.Concrete steps are as follows:
(1) respectively with the genomic dna of the gene pool of two parents and different seed coat colour for template, the primer respectively in employing table 2 carries out pcr amplification, obtains pcr amplification product respectively.
PCR reaction system is that 5 μ L (100ng) genomic DNA template, 2 μ L10 × PCRbuffer are (containing Mg
2+), 1.5 μ L2mmolL
-1dNTP, 1.5 μ L2mmolL
-1forward and reverse primer, 0.2 μ L 5UL
-1taq DNA polymerase, 8.1 μ L ddH
2o complements to 20 μ L.
PCR response procedures is: 95 DEG C of denaturation 5min, 94 DEG C of sex change 30s, peculiar annealing temperature (SSR primer the is all 47 DEG C) 30s of often pair of primer, and 72 DEG C extend 30s, 35 circulations, and 72 DEG C extend 8min, 4 DEG C of preservations.Other type primer program sets according to primer self-characteristic.
Table 2, the forward and reverse primer screened for BSA
(2) in pcr amplification product, add 5 μ l10 × LoadingBuffer, 95 DEG C of sex change 3-5min, taking-up is placed on mixture of ice and water and cools rapidly.With 6% denaturing polyacrylamide gel, amplified production is separated, on electrophoresis, tank liquor is 0.3 × TBE, lower tank liquor is 1 × TBE, the DNA cloning product 2 μ L after sex change is clicked and entered after firm power 100W prerunning is about 30min, electrophoresis is about 1h, and electrophoresis time adjusts slightly depending on the difference size between the base fragment length of SSR amplified production and parent.After electrophoresis terminates, sheet glass is put into stationary liquid (1350mlRO water+150ml dehydrated alcohol+9ml Glacial acetic acid) and fix 10-20 minute; The sheet glass fixed is put into silver-colored dye liquor and dyes 10-20 minute (silver-colored dye liquor: 1500mlRO water is containing 3g Silver Nitrate); Afterwards sheet glass from the rapid taking-up washing bath, put into developing solution to DNA band and show (developing solution: 1200ml distilled water containing sodium hydroxide 18g, the methyl ether 6ml of 37%); Finally taking out sheet glass use water and rinse removal developing solution, takes a picture and adds up banding pattern in drying at room temperature railway carriage or compartment.
Found by the screening of BSA method: the mark being positioned at G, T, I, R location proximate on No. 1 karyomit(e), No. 6 karyomit(e)s, No. 8 karyomit(e)s, No. 9 karyomit(e)s is closely related with being separated of seed coat colour.What wherein control yellow black, yellowish-brown, green black, green and brown separation is I site, and what control greenish-yellow separation is G site, and what control dark brown separation is T and R site.
Four, the acquisition of Fine Mapping and compact linkage molecule mark
The material that all 3:1 are separated is identified respectively with the mark of 4 location proximate, in these materials, screening exchanges the material of restructuring, and carry out mark encryption nearby, interval interior all polymorphism mark qualifications are utilized to exchange recombined material, and namely qualification exchange recombined material offspring separation case isozygotys or heterozygosis, search out mark closely linked with it, determine between positioning area.
By Fine Mapping, scope and the molecule marker thereof of location, G, T, I, R site are as follows:
1, I site
I site is positioned at the scope interior (Fig. 5) of 291kb between 08_459 and 08_875.Obtain in the positioning area to be divided into soybean I site from SSR molecular marker, called after SSRI, SSRI are made up of upstream primer SSRI-F and downstream primer SSRI-R.Wherein, the sequence of upstream primer SSRI-F is 5 '-TGACATGGCGTGATTGTTCT-3 ' (sequence 1), and the sequence of downstream primer SSRI-R is 5 '-ACACATCCAGTCCCAGGAAA-3 ' (sequence 2).
2, G site
In the scope that G site is positioned at 213kb between 01_1523 and 01_1536 (Fig. 3), obtain in positioning area to be divided into soybean G site from SSR molecular marker, called after SSRG, SSRG are made up of upstream primer SSRG-F and downstream primer SSRG-R.Wherein, the sequence of upstream primer SSRG-F is 5 '-TCTTAAGATTTGCCCCGTTG-3 ' (sequence 3), and the sequence of downstream primer SSRG-R is 5 '-GGCTTTTACAAAATATTGAAGTTTGA-3 ' (sequence 4).
3, R site
R site is positioned at 338kb between 09_1480 and 09_1503 (Fig. 4), obtain in the positioning area to be divided into soybean R site from SSR molecular marker, called after SSRR, SSRR are made up of upstream primer SSRR-F and downstream primer SSRR-R.Wherein, the sequence of upstream primer SSRR-F is 5 '-TTGCTTGTACCTGGCAATCA-3 ' (sequence 5), and the sequence of downstream primer SSRR-R is 5 '-TTTGTTCCTTTGGTTTTCTTTCA-3 ' (sequence 6).
4, T site
T site is positioned between 06_0994 and 06_1002, obtain in the positioning area to be divided into soybean T site from SSR molecular marker, called after SSRT, SSRT are made up of upstream primer SSRT-F and downstream primer SSRT-R.Wherein, the sequence of upstream primer SSRT-F is 5 '-TCAAAACAATAATTTTCGAGGAA-3 ' (sequence 7), and the sequence of downstream primer SSRT-R is 5 '-TGTAATTATTTTGAGTCGAAGGAAG-3 ' (sequence 8).
In sum, SSRI, SSRG, SSRR and SSRT is respectively with 4 closely linked SSR marker of site I, G, R and T.
The detection application in F2 colony of embodiment 2, molecule marker
Utilize 4 closely linked SSR marker SSRI in site, the genotype of 171 all individual plants of F2 colony that step 2 that SSRG, SSRR and SSRT identify embodiment 1 obtains, in F2 colony, total kernel seed coat colour is that green soybean has 109, kernel seed coat colour is that yellow soybean has 30, kernel seed coat colour is that the soybean of black has 15, kernel seed coat colour is that the soybean of brown has 17.Concrete steps are as follows:
Respectively with the genomic dna of 171 all individual plants of F2 colony for template, molecule marker SSRI, SSRR, SSRT and SSRG is adopted to carry out pcr amplification respectively, obtain pcr amplification product respectively, pcr amplification product is carried out denaturing polyacrylamide electrophoresis, and pcr amplification product is checked order.
PCR reaction system is that 5 μ L (100ng) genomic DNA template, 2 μ L10 × PCRbuffer are (containing Mg
2+), 1.5 μ L2mmolL
-1dNTP, 1.5 μ L2mmolL
-1forward and reverse primer, 0.2 μ L 5UL
-1taq DNA polymerase, 8.1 μ L ddH
2o complements to 20 μ L.
PCR response procedures is: 95 DEG C of denaturation 5min, 94 DEG C of sex change 30s, peculiar annealing temperature (SSR primer the is all 47 DEG C) 30s of often pair of primer, and 72 DEG C extend 30s, 35 circulations, and 72 DEG C extend 8min, 4 DEG C of preservations.Other type primer program sets according to primer self-characteristic.
Result is as shown in table 3 and Fig. 6: as can be seen from Table 3:
1) in 171 F2 colonies, 109 parts of green kind of leather materials are 224bp in the pcr amplification product size that molecule marker SSRI is corresponding, the pcr amplification product size corresponding at molecule marker SSRG is 258bp, being 248 or 288bp at the pcr amplification product that molecule marker SSRR is corresponding, is 294bp or 300bp at the pcr amplification product that molecule marker SSRT is corresponding;
2) 30 parts of yellow race's leather materials are 224bp in the pcr amplification product size that molecule marker SSRI is corresponding, the pcr amplification product size corresponding at molecule marker SSRG is 270bp, being 248 or 288bp at the pcr amplification product that molecule marker SSRR is corresponding, is 294bp or 300bp at the pcr amplification product that molecule marker SSRT is corresponding;
3) 15 parts of Black seed coat materials are 186bp in the pcr amplification product size that molecule marker SSRI is corresponding, be 258bp or 270bp in the pcr amplification product size that molecule marker SSRG is corresponding, the pcr amplification product size corresponding at molecule marker SSRR is 288bp, and the pcr amplification product size corresponding at molecule marker SSRT is 300bp;
4) 17 parts of brown seed coat color materials are divided into the following two kinds situation: wherein 15 parts of brown seed coat color materials are be 186bp in the pcr amplification product size that molecule marker SSRI is corresponding, are 258bp or 270bp in the pcr amplification product size that molecule marker SSRG is corresponding; The pcr amplification product size corresponding at molecule marker SSRR is 248bp, and the pcr amplification product size corresponding at molecule marker SSRT is 300bp; Wherein 2 parts of brown seed coat color materials are be 186bp in the pcr amplification product size that molecule marker SSRI is corresponding, be 258bp or 270bp in the pcr amplification product size that molecule marker SSRG is corresponding, be 248bp or 288bp in the pcr amplification product size that molecule marker SSRR is corresponding, the pcr amplification product size corresponding at molecule marker SSRT is 294bp.
Illustrate that colony's mesosperm look can make a distinction by of the present invention molecule marker SSRI, SSRG, SSRR and SSRT completely.
The corresponding relation of the amplified production in table 3, soybean seed coat color and 4 sites
The genotype identification result of each individual plant of F2 colony shows: molecule marker SSRI, SSRR, SSRT and SSRG can be utilized to identify soybean seed coat color as follows:
If the pcr amplification product size of primer pair 1 correspondence is 224bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp or 288bp, and the pcr amplification product size of primer pair 4 correspondence is 294bp or 300bp, then soybean seed coat color to be measured is green;
If the pcr amplification product size of primer pair 1 correspondence is 224bp, and the pcr amplification product size of primer pair 2 correspondence is 270bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp or 288bp, and the pcr amplification product size of primer pair 4 correspondence is 294bp or 300bp, then soybean seed coat color to be measured is yellow;
If the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp or 270bp, and the pcr amplification product size of primer pair 3 correspondence is 288bp, and the pcr amplification product size of primer pair 4 correspondence is 300bp, then soybean seed coat color to be measured is black;
If the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp or 270bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp, and the pcr amplification product size of primer pair 4 correspondence is 300bp; Or the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp or 270bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp or 288bp, and the pcr amplification product size of primer pair 4 correspondence is 294bp, then soybean seed coat color to be measured is brown.
Claims (10)
1. a primer for detection or auxiliary detection soybean seed coat color proterties to be measured, is following 1)-4):
1) primer set be made up of primer pair 1, primer pair 2, primer pair 3 and primer pair 4:
2) primer pair 1;
3) primer set be made up of primer pair 1 and primer pair 2;
4) primer set be made up of primer pair 1, primer pair 3 and primer pair 4;
Described primer pair 1 is for being made up of the single strand dna shown in the single strand dna shown in sequence 1 and sequence 2;
Described primer pair 2 is for being made up of the single strand dna shown in the single strand dna shown in sequence 3 and sequence 4;
Described primer pair 3 is for being made up of the single strand dna shown in the single strand dna shown in sequence 5 and sequence 6;
Described primer pair 4 is for being made up of the single strand dna shown in the single strand dna shown in sequence 7 and sequence 8.
2. a PCR reagent for detection or auxiliary detection soybean seed coat color proterties to be measured, comprises primer according to claim 1.
3. PCR reagent according to claim 2, is characterized in that: described primer pair 1, described primer pair 2, described primer pair 3 and the final concentration of described primer pair 4 in described PCR reagent are 0.2 μm of ol/L.
4. a test kit for detection or auxiliary detection soybean seed coat color proterties to be measured, comprises the PCR reagent described in primer according to claim 1 or Claims 2 or 3.
5. primer according to claim 1 or the PCR reagent described in Claims 2 or 3 or test kit according to claim 4 are in the application detected or in auxiliary detection soybean seed coat color proterties to be measured.
6. primer according to claim 1 or the PCR reagent described in Claims 2 or 3 or test kit according to claim 4 preparation detect or auxiliary detection soybean seed coat color proterties to be measured product in application.
7. primer according to claim 1 or the PCR reagent described in Claims 2 or 3 or test kit according to claim 4 or the application described in claim 5 or 6, is characterized in that: described soybean seed coat color proterties is green, yellow, black or brown.
8. a method for detection or auxiliary detection soybean seed coat color proterties to be measured is following (A)-(D):
(A) comprise the steps:
A1) by 2 in claim 1) described in primer pair 1 pcr amplification is carried out to the genomic dna of soybean to be measured, obtain pcr amplification product;
A2) size of described pcr amplification product is detected,
If the pcr amplification product size of primer pair 1 correspondence is 224bp, then soybean seed coat color to be measured be or candidate for green or yellow;
If the pcr amplification product size of primer pair 1 correspondence is 186bp, then soybean seed coat color to be measured is or candidate is black or brown;
B) comprise the steps:
B1) 3 in claim 1 is used respectively) described primer pair 1 and primer pair 2 carry out pcr amplification to the genomic dna of soybean to be measured, obtains 2 kinds of pcr amplification products;
B2) size of described 2 kinds of pcr amplification products is detected,
If the pcr amplification product size of primer pair 1 correspondence is 224bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp, then soybean seed coat color to be measured be or candidate for green;
If the pcr amplification product size of primer pair 1 correspondence is 224bp, and the pcr amplification product size of primer pair 2 correspondence is 270bp, then soybean seed coat color to be measured be or candidate for yellow;
C) comprise the steps:
C1) use 3 in claim 1 respectively) described in primer pair 1, primer pair 3 and primer pair 4 pcr amplification is carried out to the genomic dna of soybean to be measured, obtain 3 kinds of pcr amplification products;
C2) size of described 3 kinds of pcr amplification products is detected,
If the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 3 correspondence is 288bp, and the pcr amplification product size of primer pair 4 correspondence is 300bp, then soybean seed coat color to be measured is or candidate is black;
If the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp, and the pcr amplification product size of primer pair 4 correspondence is 300bp,
Or the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp or 288bp, and the pcr amplification product size of primer pair 4 correspondence is 294bp, then soybean seed coat color to be measured is or candidate is brown;
D) comprise the steps:
D1) use 4 in claim 1 respectively) described in primer pair 1, primer pair 2, primer pair 3 and primer pair 4 pcr amplification is carried out to the genomic dna of soybean to be measured, obtain 4 kinds of pcr amplification products;
D2) size of described 4 kinds of pcr amplification products is detected,
If the pcr amplification product size of primer pair 1 correspondence is 224bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp or 288bp, and the pcr amplification product size of primer pair 4 correspondence is 294bp or 300bp, then soybean seed coat color to be measured be or candidate for green;
If the pcr amplification product size of primer pair 1 correspondence is 224bp, and the pcr amplification product size of primer pair 2 correspondence is 270bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp or 288bp, and the pcr amplification product size of primer pair 4 correspondence is 294bp or 300bp, then soybean seed coat color to be measured be or candidate for yellow;
If the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp or 270bp, and the pcr amplification product size of primer pair 3 correspondence is 288bp, and the pcr amplification product size of primer pair 4 correspondence is 300bp, then soybean seed coat color to be measured is or candidate is black;
If the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp or 270bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp, and the pcr amplification product size of primer pair 4 correspondence is 300bp; Or the pcr amplification product size of primer pair 1 correspondence is 186bp, and the pcr amplification product size of primer pair 2 correspondence is 258bp or 270bp, and the pcr amplification product size of primer pair 3 correspondence is 248bp or 288bp, and the pcr amplification product size of primer pair 4 correspondence is 294bp, then soybean seed coat color to be measured is or candidate is brown.
9. method according to claim 8, is characterized in that: the annealing temperature of described pcr amplification is 47 DEG C.
10. primer according to claim 1 or the PCR reagent described in Claims 2 or 3 or test kit according to claim 4 or the application of the method described in claim 8 or 9 in Soybean Germplasm qualification and/or soybean breeder.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610069040.0A CN105543386B (en) | 2016-02-01 | 2016-02-01 | A kind of molecular labeling and its application with soybean seed coat color close linkage |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610069040.0A CN105543386B (en) | 2016-02-01 | 2016-02-01 | A kind of molecular labeling and its application with soybean seed coat color close linkage |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105543386A true CN105543386A (en) | 2016-05-04 |
| CN105543386B CN105543386B (en) | 2019-01-08 |
Family
ID=55822943
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610069040.0A Active CN105543386B (en) | 2016-02-01 | 2016-02-01 | A kind of molecular labeling and its application with soybean seed coat color close linkage |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105543386B (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111334602A (en) * | 2020-03-16 | 2020-06-26 | 中国农业科学院作物科学研究所 | Molecular marker for soybean branch number and application thereof |
| CN111663000A (en) * | 2020-07-06 | 2020-09-15 | 中国农业科学院作物科学研究所 | Soybean locked flower molecular marker and application thereof |
| CN112094334A (en) * | 2020-10-20 | 2020-12-18 | 中国农业科学院作物科学研究所 | Application of GmDCL2a protein and GmDCL2b protein in regulating and controlling soybean seed coat color |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1814809A (en) * | 2005-12-16 | 2006-08-09 | 沈阳农业大学 | RAPD label related to soybean yellow seed gene and its obtaining method and use |
| KR100707832B1 (en) * | 2005-11-16 | 2007-04-13 | 한국생명공학연구원 | Molecular marker for the seed coat color of soybean |
-
2016
- 2016-02-01 CN CN201610069040.0A patent/CN105543386B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR100707832B1 (en) * | 2005-11-16 | 2007-04-13 | 한국생명공학연구원 | Molecular marker for the seed coat color of soybean |
| CN1814809A (en) * | 2005-12-16 | 2006-08-09 | 沈阳农业大学 | RAPD label related to soybean yellow seed gene and its obtaining method and use |
Non-Patent Citations (2)
| Title |
|---|
| Q.J.SONG ET AL.: "A new integrated genetic linkage map of the soybean", 《THEORETICAL & APPLIED GENETICS》 * |
| 王吴彬等: "大豆结荚习性、荚色和种皮色相关野生片段分析", 《作物学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111334602A (en) * | 2020-03-16 | 2020-06-26 | 中国农业科学院作物科学研究所 | Molecular marker for soybean branch number and application thereof |
| CN111663000A (en) * | 2020-07-06 | 2020-09-15 | 中国农业科学院作物科学研究所 | Soybean locked flower molecular marker and application thereof |
| CN112094334A (en) * | 2020-10-20 | 2020-12-18 | 中国农业科学院作物科学研究所 | Application of GmDCL2a protein and GmDCL2b protein in regulating and controlling soybean seed coat color |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105543386B (en) | 2019-01-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109706261B (en) | Method for identifying authenticity of watermelon variety and special SNP primer combination thereof | |
| CN109251995B (en) | CAPS molecular marker for identifying watermelon peel background color and application thereof | |
| CN109762921B (en) | SNP (Single nucleotide polymorphism) marker for detecting color of cucumber pulp and application thereof | |
| CN108034752B (en) | InDel molecular labelings with eggplant fruit color epistatic gene P close linkages and application | |
| CN110229928A (en) | Molecular labeling combination and its application for Eggplant Germplasm Resources identification | |
| CN105543386A (en) | Molecular marker tightly interlocked with soybean seed coat color and application of molecular marker | |
| CN108486276A (en) | Capsicum maturity SNP marker and its application | |
| CN110438252A (en) | Molecular labeling and its application with spinach male and female gender close linkage | |
| CN109207622A (en) | A kind of molecular labeling and application with the stagnant green gene linkage of capsicum | |
| CN107142324A (en) | Mulberry tree EST SSR molecular markers and its core primers group and application | |
| CN104926931A (en) | Female sterile gene of paddy rice and application of female sterile gene | |
| CN110205396B (en) | SNP marker closely linked with irregular stripe character of cucumber fruit and application thereof | |
| CN107586874B (en) | Primer pair for identifying yellow anther character of capsicum and application thereof | |
| CN114015796A (en) | Molecular marker closely linked with control of light white peel of American pumpkin, primer and application | |
| CN105695454B (en) | A kind of molecular labeling and its identification method for identifying sesame genie male sterile line | |
| CN109852723B (en) | SNP molecular marker closely linked with cowpea pod color gene and application thereof | |
| CN109609676B (en) | SNP (Single nucleotide polymorphism) marker co-separated from corn gray spot resistant major QTL-qRgls1 and application | |
| CN111088383B (en) | Molecular marker for identifying purple genes of capsicum olivum and development method and application thereof | |
| CN106434906B (en) | A kind of and cauliflower bouquet hair floral formation chain SNP site and its detection method and primer sets | |
| CN108085404A (en) | Giant pumpkin female trait molecular marker and the primer pair of identification giant pumpkin female character by force by force | |
| CN105524993B (en) | The molecular labeling HRM1 of barley grain length gene Lkl1 a kind of and its application | |
| CN114622033B (en) | SNP (Single nucleotide polymorphism) marker linked with color traits of ripe cucumber peel and application thereof | |
| CN105603068B (en) | The molecular labeling of soybean plant height close linkage and its application | |
| CN105087802B (en) | The method of the identification anti-late blight character of tomato and its molecular labeling used and primer | |
| CN110106270A (en) | The molecular labeling and its application that a kind of and muskmelon yellow seed coat isolates |
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 | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240219 Address after: 102200, 7th floor, Building 4, No. 4 Life Park Road, Zhongguancun Life Science Park, Changping District, Beijing Patentee after: BEIJING COMPASS BIOTECHNOLOGY TECHNOLOGY Co.,Ltd. Country or region after: China Address before: 100081 Institute of crop science, Chinese Academy of Agricultural Sciences, 12 South Avenue, Beijing, Haidian District, Zhongguancun Patentee before: INSTITUTE OF CROP SCIENCES, CHINESE ACADEMY OF AGRICULTURAL SCIENCES Country or region before: China |