CN106755478B - SSR molecular marker IV for identifying progeny plants of Gala apples and application thereof - Google Patents
SSR molecular marker IV for identifying progeny plants of Gala apples and application thereof Download PDFInfo
- Publication number
- CN106755478B CN106755478B CN201710040750.5A CN201710040750A CN106755478B CN 106755478 B CN106755478 B CN 106755478B CN 201710040750 A CN201710040750 A CN 201710040750A CN 106755478 B CN106755478 B CN 106755478B
- Authority
- CN
- China
- Prior art keywords
- apple
- gala
- ssr molecular
- molecular marker
- plant
- 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.)
- Expired - Fee Related
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/6895—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for plants, fungi or algae
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Analytical Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Molecular Biology (AREA)
- Physics & Mathematics (AREA)
- Mycology (AREA)
- Botany (AREA)
- Immunology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention belongs to the field of plant genetic breeding and apple germplasm innovation research, and particularly relates to an SSR molecular marker IV for identifying a gala apple progeny plant and application thereof. Identifying the interlocked apple groups where the identified materials are located by using SSR molecular markers for the first time, and simultaneously disclosing SSR molecular markers interlocked with No. 5 and No. 10 chromosomes of the apples; the molecular marker is a codominant marker, and can quickly and accurately identify the Gala apple genetic linkage group, the anther culture plant and the Gala source plant; and the molecular level support is provided for accelerating the utilization of the Gala apple and important agronomic character linked genes and the genetic breeding of apple homozygous plants in the next step.
Description
Technical Field
The invention belongs to the field of plant genetic breeding and apple germplasm innovation research, and particularly relates to an SSR molecular marker IV for identifying a gala apple progeny plant and application thereof.
Background
Apple (A)Malus domesticaBorkh.) has strong ecological adaptability and high nutritive value of fruits, and is one of the fruit tree species with wide cultivation area, large consumption and better economic benefit in the world. China is one of the countries with the widest apple cultivation area and the highest total output. Particularly in northern China, apples are the fruit tree species with the largest cultivation area, and the yield and the area of the apples are the first fruits in China. The industry becomes a post industry of many provinces and cities in China, and plays more and more important roles in improving income of farmers and promoting development of local economy. Apples belong to self-flowering infertile plants, apple varieties in production are heterozygous diploid varieties, apple genomes are highly heterozygous, genetic backgrounds are quite complex, and the childhood period is long, so that the conventional cross breeding period is long and the efficiency is low.
The breeding efficiency of the homozygous genotype germplasm can be greatly improved. Haploid plants can be obtained by anther culture, and homozygous diploid germplasm can be quickly obtained after chromosome doubling. Since apples are highly heterozygous diploid varieties, they are usually controlled by multiple alleles at a single locus, i.e. the same locus contains two different alleles, whereas haploid varieties contain only one gene. The plant obtained by anther culture should have only one allele of one of its parents if it is of haploid origin. Homozygous diploid germplasm can be obtained through anther culture, and recessive gene materials and mutation breeding materials with excellent characters can be directly obtained, and the materials have important significance for apple genetic breeding research.
Molecular marker technology has been applied in early selection of major agronomic traits of apple, such as red flesh, red skin, scab, woolly apple aphid, fire blight, etc., participating in completing the apple Genome sequencing work and jointly developing 8K (CHAGN É D, etc., Genome-wide SNP detection, identification, and identification of an 8K SNP array for apple [ J ]. P L oS ONE, 2012, 7: e31745. doi: 10.1371/journel.point.0031745) and 20K (ANCBIO L, etc., Development and identification of a 20K Single Nucleotide Polymorphism (SNP) Genome for apple (Malus × Genome) [ J ]. P2, GS 2014: 1109: 10 cross breeding, 11064, SNP breeding of apple using the Bolus × Genome genomic PCR strain [ J ] 387 [ 10 ] SNP (SNP) in stead of the apple Genome breeding program (GS 10.1371: 0364) in breeding.
Compared with other molecular markers such as RF L P, AF L P ISSR and the like, the SSR marker has the characteristics of high polymorphism, co-dominant inheritance, good repeatability, strong specificity and the like, and becomes a marker which is most applied in the fields of genetic diversity research, genetic mapping, important functional gene positioning, molecular assisted breeding and the like in recent years.
SSR markers are widely used in fruit quality trait marker screening, variety Identification and genetic linkage map construction due to their good stability and transferability (L iu, et al. Identification of apple cucumber orientations of simple sequence repeat markers, Genet Mol Res, 2014, 13 (3): 7377 7387; Moriya, et al. Aligned genetic linkage maps of apple rootstock culture 'JM 7' and Malus sieboldii 'Sanashi 63' constrained with novel EST-SSRs. Tree Genet genes, 2012, 8 (4): 709 723.).
So far, no SSR molecular marker of the Gala apple anther culture plant is reported.
Disclosure of Invention
The invention aims to provide an SSR molecular marker IV for identifying the progeny plants of Gala apples and application thereof.
The invention is realized by the following technical scheme: an SSR molecular marker IV for identifying the progeny plants of the Gala apples is used simultaneously in the detection process, and the molecular marker is 2 pairs of SSR primers with nucleotide sequences shown as follows:
l G5 marker Hi04d 02:
upstream: 5，- TTCGTGGCTGAGAAAGGAGT -3，
Downstream: 5，- GTTTGTACGGTGCATTGTGAAAG -3，
L G10 marker Hi05b 02:
upstream: 5，- GATGCGGTTTGACTTGCTTC -3，
Downstream: 5，- GTTTCTCCAGCTCCCATAGATTGC -3，。
An application of an SSR molecular marker IV for identifying the progeny plants of Gala apples comprises the following steps:
(1) the Gala apple genome DNA is used as a PCR amplification template, the SSR molecular markers are respectively used as primer pairs to carry out PCR amplification, the reaction system is 15 mu L, wherein the reaction system comprises 10 × PCR Buffer1.5 mu × 0, 2.5 mM dNTPs mix 1.2 mu × 1, 10 ng/mu L Primers F1.5 mu L, 10 ng/mu L Primers R1.5 mu L, 5U Taq polymerase 0.15 mu L, 100 ng/mu L DNA template 0.75 mu L, deionized water is supplemented to 15 mu L, and the amplification program (2) is that the PCR amplification program is carried out at 94 ℃ for 2 min 30 s, the PCR amplification program is carried out at 94 ℃ for 30 s, the PCR amplification program is carried out at 60 ℃ for 30 s, the PCR amplification program is carried out at 72 ℃ for 40s, and the PCR amplification program is carried out for 35 cyclesDetecting PCR products, performing 8% non-denaturing polyacrylamide electrophoresis, adding each reaction product of PCR into 1/2 non-denaturing L oadding buffer, mixing, fixing with 150V constant voltage for 150min, and AgNO3Dyeing and photographing; (4) when the method is used for genetic linkage identification, if any specific strip corresponding to the 2 pairs of SSR primers can be amplified, the genetic material contained in the to-be-detected apple germplasm is located in the corresponding genetic linkage group, otherwise, the genetic material contained in the to-be-detected apple germplasm does not have the corresponding genetic linkage group; (5) when the kit is used for identifying anther culture plants, the apple to be detected can amplify a single specific strip corresponding to the 2 pairs of SSR primers, so that the apple plant to be detected is a homozygous material, and if two strips appear, the apple plant to be detected is not the homozygous material; (6) when the method is used for variety source identification, the apple to be detected can amplify any one specific strip corresponding to the 2 pairs of SSR primers, which indicates that the apple plant to be detected is from the Gala apple, and if no specific strip is amplified, the apple plant to be detected is not a progeny variety cultured by the Gala apple.
Compared with the prior art, the invention has the following advantages:
1. the invention reports that SSR molecular markers are used for identifying the linkage group of apples where the identification material is located for the first time at home and abroad, and simultaneously reports the SSR molecular markers linked on No. 5 chromosomes and No. 10 chromosomes of apples;
2. the molecular marker is a co-dominant marker, and can quickly and accurately identify the Gala apple genetic linkage group, the anther culture plant and the Gala breeding progeny variety;
3. the research results provide a molecular level support for accelerating the utilization of the Gala apple and important agronomic character linked genes and the genetic breeding of the apple homozygous plants in the next step;
4. provides a method for quickly identifying the molecular level verification of the Gala cultivated plants.
By identifying 2 pairs of SSR molecular markers linked on chromosomes 5 and 10 of the flower drug cultured plant of the Gala apple, the genotype type and the genetic diversity of the Gala apple can be systematically and accurately known, a foundation is laid for enriching and developing an apple allele system, a theoretical basis is provided for scientific utilization of the Gala apple in future, and the genotype and the homozygosity of the flower drug cultured plant of the Gala apple are identified. The invention has positive promoting effect on the breeding of new apple varieties and molecular marker-assisted breeding. Meanwhile, the 2 pairs of SSR molecular markers screened by the method also provide support for source verification of the progeny varieties of the Gala apples on the molecular level.
Drawings
FIG. 1 is a capillary electrophoresis pattern of L G5 labeled Hi04d02, and FIG. 2 is a pattern of results of non-denaturing polyacrylamide gel electrophoresis detection of L G10 labeled Hi05b 02.
Detailed Description
The homozygous genotype has important roles In higher Plant genetic mechanism research and Breeding application (Murovec, et al. Haploids and double Haploids In Plant Breeding. In: Abdurakhmonov I (ed) Plant Breeding: 2012: 87-106.). Apple is a fruit tree species with a highly heterozygous genome, and the difficulty of genome assembly can be greatly reduced by using a haploid genome (Dunwell. Haploids aerating plants: origins and ex-ploitation. Plant Biotechnol J, 2010, 8 (4): 377-424.). The apple is a perennial herb, the reproductive cycle is long, and the self-incompatibility causes that the method for obtaining the homozygous plant through multi-generation self-crossing is difficult to realize. The induction of embryoid production by anther Culture to obtain homozygous genotype lines is of great significance for breeding and genetic analysis of apples with highly heterozygous genotypes (German. economic embryo and dhaproid technology as available competent to Plant breeding. Plant Cell Rep,2011, 30(5): 839-. The regenerated plant is obtained by inducing embryoid through anther culture, and the ploidy and the source of the obtained regenerated plant are accurately identified, so that the method has important significance for innovative germplasm genetic analysis. In order to better utilize the germplasm materials, the invention identifies the linkage group and the genotype of the Gala apple and the plant obtained by the anther culture of the Gala apple, and provides a molecular level support for accelerating the utilization of the Gala apple and the important agronomic character linkage gene and the genetic breeding of the apple homozygous plant.
Gala apple flower medicine culture
After the Gala apples have buds in the last ten days of 4 months, selecting the mature Gala apples by a mixed sampling method, collecting well-developed buds, sealing the buds by a sealing bag, and placing the buds in a refrigerating chamber at 4 ℃ of a refrigerator for low-temperature pretreatment. After low-temperature treatment, sterilizing the flower buds in an ultra-clean workbench by using 0.1% sodium hypochlorite, taking out the anthers from the flower buds by using a pair of tweezers, inoculating the anthers to an embryoid induction culture medium, carrying out dark culture at 25 ℃, transferring the embryoids to a regeneration culture medium for regeneration after 3-5 months until the embryoids grow to 8-10 mm, carrying out subculture, carrying out rooting culture, domestication and indoor transplantation to obtain regenerated plants.
SSR molecular marker analysis
Total DNA extraction of genome
Selecting 6 plants of the Gala apple and Gala flower cultured regeneration plants, and respectively extracting the total DNA of the anther by adopting a CTAB method (Caokifen et al, 2003). 2 pairs of SSR primers distributed on No. 5 and No. 10 chromosomes of the apples are selected from the high-density microsatellite genetic map of the apples constructed by a HiDRAS website and Okada and the like, and the HIDRAS markers of the linkage groups of the regenerated plants are shown in Table 1; the primers were synthesized by Biotechnology engineering (Shanghai) Inc.
Table 1:
PCR amplification
The total volume of the PCR reaction system is 15 mu L, 10 × PCR Buffer1.5 mu × 0, 2.5 mM dNTPs1.2 mu × 1, 10 ng/mu L Primers F1.5 mu L, 10 ng/mu L Primers R1.5 mu L, 5U Taq polymerase 0.15 mu L, 100 ng/mu L DNA template 0.75 mu L are contained in the PCR reaction system, deionized water is added to 15 mu L, the amplification program is pre-denatured at 94 ℃ for 2 min 30 s, denatured at 94 ℃ for 30 s, annealed at 60 ℃ for 30 s, extended at 72 ℃ for 40s, and is circulated for 35 times, and stored at 72 ℃ for 10 min and 4 ℃ for later use.
PCR product detection
And (3) performing 8% non-denaturing polyacrylamide electrophoresis, adding 1/2 non-denaturing L oadingBuffer into each reaction product of the PCR, uniformly mixing, keeping the constant voltage at 150V for about 150min, fixing the mixture by glacial acetic acid, and performing AgNO3 staining and photographing.
The method comprises the steps of screening bands obtained by 8% of non-denatured polyacrylamide electrophoresis, analyzing markers for generating polymorphism in Gala and anther culture plants thereof, counting, selecting a primer for amplifying bands of two alleles/loci in Gala apples and only one allele/locus in the Gala apple anther culture plants, amplifying products of 220 bp and 232 bp in a primer Hi09b04 (L G5), amplifying products of 200 bp and 250 bp in a primer Hi04d02 (L G5), amplifying products of 170 bp and 121 bp in a primer Hi05b02 (L G10), and screening 2 pairs of SSR molecular markers distributed on the 5 th chromosome and the 10 th chromosome to amplify allele loci in the Gala and anther culture plants thereof.
And (3) chromosome ploidy analysis of a regeneration line, namely cutting the leaves of a regeneration plant in 500 mu L lysate by using a blade, standing for 2 min, filtering in an EP tube, dyeing PI, analyzing the DNA content in the leaves by using a BD Accuri C5 flow cytometer, and taking the leaves of the heterozygous diploid Gala test-tube plantlet obtained by stem tip culture as a reference standard for ploidy identification.
And (3) carrying out chromosome ploidy identification on the survival and regeneration strain by adopting a flow cytometer. The heterozygous diploid Gala donor is taken as a control, and the result shows that: the regeneration lines of Gala 1-Gala 5 are all diploid.
Genotyping of the regenerating lines:
extraction of genomic DNA: after total DNA of leaves is extracted by adopting an improved CTAB method, the concentration of the DNA is detected by a nucleic acid protein instrument (Bio-Rad), and the concentration and the quality of the DNA are detected by 1 percent agarose gel electrophoresis.
The PCR reaction system is 25 mu L, and contains dNTP mix (10 mM) 0.5 mu L and 10 × PCR Buffer 2.5 mu L and 25 mu L mM MgCl22.0 mu L, rTaq enzyme (5U/. mu. L) 0.2 mu. L, DNA template (100 ng/. mu. L) 1 mu. L, deionized water 17.8 mu. L. the PCR reaction program comprises the first step of pre-denaturation at 95 ℃ for 3min, the second step of denaturation at 94 ℃ for 30 s, annealing at 60 ℃ for 30 s, and extension at 72 ℃ for 30 s, 10 cycles, the third step of denaturation at 95 ℃ for 30 s, annealing at 55 ℃ for 30 s, and extension at 72 ℃ for 30 s, 20 cycles, and the fourth step of sufficient extension at 72 ℃ for 6 min, and storage at 4 ℃.
The results of SSR identification of the Gala regenerated plant genotype are shown in Table 2, and the results show that 2 SSR markers on the linkage group show that a heterozygote is two peaks, while a regenerated strain only has one peak. It is proved that the Gala regeneration line Gala 1-Gala 5 is a homozygous genotype plant.
And (3) carrying out plant observation on a regeneration line: and (5) carrying out the phytology characteristic survey on the test-tube plantlet of the regenerated plant. The plant observation results of the regenerated plants are shown in table 3, and the results in table 3 show that the gala heterozygous donor plant height is 5.67 cm (n = 1) and the average plant height of the homozygous diploid is 2.96 cm ± 0.44 cm (n = 28). We also observed that homozygous diploid vigour was weaker relative to gala heterozygous donors. The different diploid homozygous plants also have different phytological characteristics, and the Gala5 plants are relatively high, the leaf base is widened, and the leaf tip is gradually tapered.
Table 3:
results and analysis
Haploid breeding is one of the most efficient methods to obtain dominant parent donor material. The anther wall of anther is heterozygote cell, and it is possible to induce the regeneration plant of heterozygote diploid, so the obtained diploid plant is not necessarily homozygote. Heterozygous diploids and homozygous diploids of anther-cultured regenerated plants can be distinguished by identifying alleles of the regenerated plants. Previous techniques including isozyme markers, S alleles and SSR molecular markers have all been applied to the homozygosity identification of anther regenerated plants. The invention also adopts SSR identification method. Firstly, the SSR marker (from HIDRAS database (http:// www.hidras.unimi.it /)) selected is used for carrying out PCR amplification on all regeneration plants, and the SSR marker which can distinguish the regeneration plants as homozygous is screened out. To further differentiate the genotypes of the regenerated plants, we screened SSR markers distributed in linkage groups on chromosomes 5 and 10 of apple. The SSR marker can clearly mark different plants of the Gala apple, and provides a technical index for the genotype identification of the Gala apple in the future.
The alleles implied by the parents are detectable in anther-cultured plants, but only one allele/locus is amplified in anther-cultured plants. This indicates that the plants grown from anthers are homozygous for their genotype, i.e. haploid from the parent.
Conclusion
Homozygote plants with rich ploidy can be obtained through anther culture, which provides rich test materials for developing ploidy genetic breeding research of apples in future.
The regeneration strain and the SSR identification system obtained by the invention have important significance for analyzing and identifying the Gala excellent character gene research, field grafting and crossbreeding and phenotype-genotype correlation analysis.
And the next step is combined with the whole genome sequencing result of the Gala apples, and the parents and the anther culture plants thereof are thoroughly analyzed and compared in the genome range so as to analyze the molecular mechanism of important characters (characteristics) of the Gala apples.
Sequence listing
110 institute of agricultural resources and economics, institute of biotechnology, and institute of fruit tree
SSR molecular marker IV for identifying progeny plants of Gala apples and application thereof
〈160〉4
〈210〉1
〈211〉20
〈212〉DNA
Artificial sequence of < 213 >
〈220〉
Upstream primer of Hi04d02 marked by < 223 > L G5
〈400〉1
TTCGTGGCTGAGAAAGGAGT
〈210〉2
〈211〉23
〈212〉DNA
Artificial sequence of < 213 >
〈220〉
Downstream primer of Hi04d02 marked by < 223 > L G5
〈400〉2
GTTTGTACGGTGCATTGTGAAAG
〈210〉3
〈211〉20
〈212〉DNA
Artificial sequence of < 213 >
〈220〉
Upstream primer of Hi05b02 marked by < 223 > L G10
〈400〉3
GATGCGGTTTGACTTGCTTC
〈210〉4
〈211〉24
〈212〉DNA
Artificial sequence of < 213 >
〈220〉
Downstream primer of Hi05b02 marked by < 223 > L G10
〈400〉4
GTTTCTCCAGCTCCCATAGATTGC
Claims (1)
1. The application of the SSR molecular marker IV for identifying the progeny plants of the Gala apples is characterized by comprising the following steps:
(1) apple genome DNA is used as a PCR amplification template, two pairs of Primers of SSR molecular marker IV are respectively used for PCR amplification, and the reaction system is 15 mu L, wherein the reaction system contains 10 × PCR buffer1.5 mu × 0, 2.5 mM dNTPs mix 1.2 mu × 1, 10 ng/mu L Primers F1.5 mu L, 10 ng/mu L Primers R1.5 mu L, 5U Taq polymerase 0.15 mu L, 100 ng/mu L DNA template 0.75 mu L, and deionized water is supplemented to 15 mu L;
(2) the amplification procedure was: pre-denaturation at 94 deg.C for 2 min and 30 s, denaturation at 94 deg.C for 30 s, annealing at 60 deg.C for 30 s, extension at 72 deg.C for 40s, and storing at 72 deg.C for 10 min and 4 deg.C for 35 cycles;
(3) detecting PCR product, performing 8% non-denaturing polyacrylamide electrophoresis, adding each reaction product of PCR into 1/2 non-denaturing L loading Buffer, mixing, keeping constant voltage at 150V for 150min, fixing with glacial acetic acid, and AgNO3Dyeing and photographing;
(4) when the SSR molecular marker is used for identifying anther culture plants, the apple to be detected can amplify a single specific strip corresponding to the two pairs of primers of the SSR molecular marker IV, so that the apple plant to be detected is a homozygous material, and if two strips appear, the apple plant to be detected is not the homozygous material;
the SSR molecular marker IV consists of L G5 markers Hi04d02 and L G10 markers Hi05b02, two pairs of primers of the SSR molecular marker IV are used simultaneously in the detection process, and the sequences of the primers are as follows:
l G5 marker Hi04d 02:
upstream: 5，- TTCGTGGCTGAGAAAGGAGT -3，
Downstream: 5，- GTTTGTACGGTGCATTGTGAAAG -3，
L G10 marker Hi05b 02:
upstream: 5，- GATGCGGTTTGACTTGCTTC -3，
Downstream: 5，- GTTTCTCCAGCTCCCATAGATTGC -3，。
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610849403 | 2016-09-26 | ||
| CN2016108494032 | 2016-09-26 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN106755478A CN106755478A (en) | 2017-05-31 |
| CN106755478B true CN106755478B (en) | 2020-07-17 |
Family
ID=58943462
Family Applications (6)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710040753.9A Expired - Fee Related CN106755479B (en) | 2016-09-26 | 2017-01-20 | SSR molecular marker V for identifying progeny plants of Gala apples and application thereof |
| CN201710040784.4A Expired - Fee Related CN106755481B (en) | 2016-09-26 | 2017-01-20 | SSR molecular marker VI for identifying progeny plants of Gala apples and application thereof |
| CN201710040750.5A Expired - Fee Related CN106755478B (en) | 2016-09-26 | 2017-01-20 | SSR molecular marker IV for identifying progeny plants of Gala apples and application thereof |
| CN201710040796.7A Expired - Fee Related CN106755482B (en) | 2016-09-26 | 2017-01-20 | SSR molecular marker III for identifying progeny plants of Gala apples and application thereof |
| CN201710040768.5A Expired - Fee Related CN106755480B (en) | 2016-09-26 | 2017-01-20 | SSR molecular marker I for identifying progeny plants of Gala apples and application thereof |
| CN201710040798.6A Expired - Fee Related CN106755483B (en) | 2016-09-26 | 2017-01-20 | SSR molecular marker II for identifying progeny plants of Gala apples and application thereof |
Family Applications Before (2)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710040753.9A Expired - Fee Related CN106755479B (en) | 2016-09-26 | 2017-01-20 | SSR molecular marker V for identifying progeny plants of Gala apples and application thereof |
| CN201710040784.4A Expired - Fee Related CN106755481B (en) | 2016-09-26 | 2017-01-20 | SSR molecular marker VI for identifying progeny plants of Gala apples and application thereof |
Family Applications After (3)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710040796.7A Expired - Fee Related CN106755482B (en) | 2016-09-26 | 2017-01-20 | SSR molecular marker III for identifying progeny plants of Gala apples and application thereof |
| CN201710040768.5A Expired - Fee Related CN106755480B (en) | 2016-09-26 | 2017-01-20 | SSR molecular marker I for identifying progeny plants of Gala apples and application thereof |
| CN201710040798.6A Expired - Fee Related CN106755483B (en) | 2016-09-26 | 2017-01-20 | SSR molecular marker II for identifying progeny plants of Gala apples and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (6) | CN106755479B (en) |
Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109825636A (en) * | 2019-04-04 | 2019-05-31 | 山西省农业科学院生物技术研究中心 | One kind is for identifying the SSR molecular marker and its application of Variety of Apple " red rosy clouds " finger-print |
| CN110295249B (en) * | 2019-06-11 | 2020-04-03 | 中国科学院武汉植物园 | SSR molecular marker for screening sugar content phenotype of apple fruits and application of SSR molecular marker |
| CN110491444A (en) * | 2019-08-13 | 2019-11-22 | 西北农林科技大学 | A kind of development approach and application of the molecular labeling of quick screening apple red meat character |
| CN110468228B (en) * | 2019-09-02 | 2022-05-31 | 中国农业科学院蔬菜花卉研究所 | Molecular marker for identifying segregation condition of interspecific hybrids and progeny materials A10 and C07 chromosomes of Chinese cabbages and brassica carinata |
| CN110499383B (en) * | 2019-09-02 | 2022-05-31 | 中国农业科学院蔬菜花卉研究所 | Molecular marker for identifying segregation condition of interspecific hybrids and progeny materials A02 and C02 chromosomes of Chinese cabbages and brassica carinata |
| CN111560463B (en) * | 2020-06-15 | 2022-08-26 | 山东丰沃植物研究院有限公司 | Three gala apple specific molecular markers and screening method and application thereof |
| CN111663002B (en) * | 2020-07-14 | 2022-10-11 | 福建农林大学 | SSR marker for distinguishing genetic background of No. two chromosomes between sugarcane species and application |
| CN112126699B (en) * | 2020-09-15 | 2022-03-01 | 中国农业大学 | Malus plant complete genome InDel marker genotype database and application thereof in germplasm resource specificity identification |
| CN113215300B (en) * | 2021-06-03 | 2022-08-02 | 山东大丰园农业有限公司 | 7 apple variety specific molecular markers and screening method and application thereof |
| CN117121809A (en) * | 2022-05-19 | 2023-11-28 | 南京农业大学 | Method for cultivating microspore plant of non-heading Chinese cabbage |
| CN117385084A (en) * | 2023-11-23 | 2024-01-12 | 西北农林科技大学 | Molecular marker closely linked with hardness of apple fruits, primer and application thereof |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1962212A1 (en) * | 2007-01-17 | 2008-08-27 | Syngeta Participations AG | Process for selecting individuals and designing a breeding program |
| CN102653790A (en) * | 2012-03-31 | 2012-09-05 | 中国农业科学院果树研究所 | Improved TP-M13-SSR molecular arking method of apple germplasm resource |
| CN104531844B (en) * | 2014-12-05 | 2018-09-04 | 中国农业科学院郑州果树研究所 | A kind of fruit variety differentiation and characteristic fingerprint methods of exhibiting based on SSR genotype |
-
2017
- 2017-01-20 CN CN201710040753.9A patent/CN106755479B/en not_active Expired - Fee Related
- 2017-01-20 CN CN201710040784.4A patent/CN106755481B/en not_active Expired - Fee Related
- 2017-01-20 CN CN201710040750.5A patent/CN106755478B/en not_active Expired - Fee Related
- 2017-01-20 CN CN201710040796.7A patent/CN106755482B/en not_active Expired - Fee Related
- 2017-01-20 CN CN201710040768.5A patent/CN106755480B/en not_active Expired - Fee Related
- 2017-01-20 CN CN201710040798.6A patent/CN106755483B/en not_active Expired - Fee Related
Also Published As
| Publication number | Publication date |
|---|---|
| CN106755479B (en) | 2020-07-17 |
| CN106755481B (en) | 2020-07-17 |
| CN106755478A (en) | 2017-05-31 |
| CN106755480A (en) | 2017-05-31 |
| CN106755483B (en) | 2020-07-17 |
| CN106755481A (en) | 2017-05-31 |
| CN106755483A (en) | 2017-05-31 |
| CN106755480B (en) | 2020-07-17 |
| CN106755482A (en) | 2017-05-31 |
| CN106755479A (en) | 2017-05-31 |
| CN106755482B (en) | 2020-07-17 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106755478B (en) | SSR molecular marker IV for identifying progeny plants of Gala apples and application thereof | |
| CN108411028B (en) | Specific SNP codominant molecular marker primer in rice salt-tolerant gene SKC1 gene and application | |
| CN108192990B (en) | SNP molecular marker related to watermelon peel background color and application thereof | |
| CN111926098B (en) | InDel molecular marker closely linked with epistatic gene Y of eggplant fruit color and application | |
| CN113151550B (en) | Molecular marker CmSSR02 closely linked with main effect QTL fft2 of early flowering characteristics of melons and application thereof | |
| CN113046467B (en) | SNP locus obviously associated with wheat stripe rust resistance and application thereof in genetic breeding | |
| CN104711254B (en) | The low-phosphorous responsive genes ZmARF31 of corn INDEL molecular labelings and its application | |
| CN105543366B (en) | Development and application of specific SNP codominant molecular marker in rice blast-resistant gene Pi25 gene | |
| CN114752702B (en) | Molecular marker BnCa-2C2 closely linked with rape calcium content trait QTL and application thereof | |
| CN108004236B (en) | Corn stalk rot disease-resistant molecular breeding method and application thereof | |
| CN111235305B (en) | SNP molecular markers related to lead transport coefficient of corn plants and application thereof | |
| CN111004857B (en) | Molecular marker primer of soybean branch number major QTL locus and application thereof | |
| CN108546778B (en) | SNP molecular marker for detecting powdery mildew resistance of cucumber and application thereof | |
| CN113278723A (en) | Composition for analyzing genetic diversity of Chinese cabbage genome segment or genetic diversity introduced in synthetic mustard and application | |
| CN107699630B (en) | Molecular marker linked with wheat disease-resistant gene Pm21 and application thereof in breeding | |
| CN114032330B (en) | SNP molecular marker for rapidly identifying apple anthracnose | |
| CN117248061B (en) | InDel locus related to soybean seed oil content, molecular marker, primer and application thereof | |
| CN112342309B (en) | SNP molecular marker related to cotton flower basal leaf spot character and application thereof | |
| CN114369674B (en) | SNP marker linked with Indian pumpkin short vine gene CmDw-1, primer, kit and application thereof | |
| Prasad et al. | Advancement in molecular tools of plant population genetics | |
| CN110724755B (en) | CAPS marker primer group linked with watermelon internode length and application thereof | |
| CN116024373A (en) | SNP molecular marker related to grape cold resistance and application thereof | |
| CN114807409A (en) | Molecular marker related to watermelon tendril degradation and self-capping and application thereof | |
| CN113736906A (en) | SNP locus combination for detecting tomato verticillium wilt resistance and application thereof | |
| CN116179747A (en) | Primer pair for eggplant peel color gene amplification |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20200717 Termination date: 20210120 |