CN106755482A - A kind of SSR molecular marker III for identifying Gala apple Progeny plants and its application - Google Patents
A kind of SSR molecular marker III for identifying Gala apple Progeny plants and its application Download PDFInfo
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Abstract
The invention belongs to plant genetics and breeding and apple germplasm innovation research field, a kind of specific SSR molecular marker III for identifying Gala apple Progeny plants and its application, used simultaneously in detection process, molecular labeling is SSR molecular marker chain on 4,6,12 and No. 14 chromosomes, template is expanded by PCR of apple genome DNA, respectively with SSR molecular marker as primer pair, enter performing PCR amplification and product detection, for genetic linkage identification, anther plant identification, cultivar origin identification.Apple linkage group residing for expert evidence is identified with SSR molecular marker first, is disclosed and 4,6,12 and No. 14 SSR molecular markers of chromosome linkage of apple；The molecular labeling is codominant marker, quick, accurate that Gala apple genetic linkage group, anther plant and loud, high-pitched sound source plants are identified；Accelerate Gala apple for next step and provide molecular level support with the utilization of Main Agronomic Characters linked gene and apple homozygous plants genetic breeding.
Description
Technical field
The invention belongs to plant genetics and breeding and apple germplasm innovation research field, after specially a kind of identification Gala apple
For the SSR molecular marker III of plant and its application.
Background technology
Apple（Malus domesticaBorkh.）Annidation is strong, and fruit is of high nutritive value, and is to cultivate in the world
Area is wide, consumption figure is big, one of better economic benefit fruit tree species.China is apple cultivation area most wide, total output highest
One of country.Especially in northern China, apple is the maximum fruit tree species of cultivated area, and its yield and area occupy national water
First of fruit.This industry has turned into the pillar industry of many provinces and cities of China, and they are improving farmers' income, are promoting local economy hair
More and more important effect is played in exhibition.Malus self-sterile plant, the apple variety in production is heterozygous diploid product
Kind, apple genome height heterozygosis, genetic background is sufficiently complex, and its juvenile phase is very long in addition so that the conventional cross-breeding cycle is long,
Efficiency is low.
Directive breeding efficiency can be then greatly improved using the breeding of homozygous genotype germplasm.Anther Culture can obtain single times
Body plant, the dliploid germplasm by can rapidly obtain homozygosis after chromosome doubling.Because apple is two times of height heterozygosis
Body kind, generally in single site by multiple allele control, i.e., two kinds of different allele are contained in same site, and list is again
Body kind is containing only a kind of gene.Plant is obtained by Anther Culture, if its be monoploid origin, should only have its parent it
A kind of one allele.The dliploid germplasm of homozygosis can be obtained by Anther Culture, proterties can also be directly obtained excellent
Recessive gene material and mutation breeding material, these materials are significant to apple genetic breeding research.
Molecular marking technique is applied in the Seedling selection of apple Other Main Agronomic Characters, such as red meat, red skin, black star
Disease, eriosoma lanigerum and fire blast etc., participate in completing apple genome examining order and having developed jointly 8 K（CHAGN é D etc.,
Genome-wide SNP detection, validation, and development of an 8K SNP array for
apple[J]. PLoS ONE, 2012, 7: e31745. doi:10.1371/journal.pone.0031745）With 20 K
（BIANCO L etc., Development and validation of a 20K Single Nucleotide
Polymorphism（SNP）whole genome genotyping array for apple ( Malus × domestica
Borkh) [J] PLoS One, 2014,9 (10): e110377. dio：10.1371/journal.pone.0110377）
Apple SNP chip, the 1st time in world's Apple breeding project use gene group selection（GS）Method adds instead of phenotypic screen
Fast breeding paces.
Simple sequence repeats (simple sequence repeat, SSR) are a class 1-6bp nucleotides motif structures
Into repetitive sequence, code area, the noncoding region of eukaryotic gene group are distributed widely in, SSR marks utilize simple sequence repeats
Flank conserved sequence design primer, expanded by PCR, size according to band reflects the polymorphism of DNA sequence dna.With it
Its molecular labeling such as RFLP, AFLP ISSR etc. is compared, SSR marks have that polymorphism is high, codominant inheritance, it is reproducible,
The features such as high specificity, educated as genetic diversity Journal of Sex Research, genetic mapping, the critical function assignment of genes gene mapping, molecule auxiliary in recent years
The fields such as kind are using a kind of most marks.
SSR marker has good stability and transferability because of it, in fruit quality proterties label screening, cultivar identification
It is widely used in being built with genetic linkage mapses（Liu, waits Identification of apple cultivars on
the basis of simple sequence repeat markers. Genet Mol Res, 2014, 13 ( 3) :
7377-7387;Moriya, waits Aligned genetic linkage maps of apple rootstock
cultivar ‘JM7’ and Malus sieboldii ‘Sanashi 63’ constructed with novel EST -
SSRs. Tree Genet Genomes, 2012, 8 (4) : 709-723.）.
Up to the present, the SSR molecular marker for Gala apple anther plant has not been reported.
The content of the invention
It is an object of the invention to provide a kind of SSR molecular marker III for identifying Gala apple Progeny plants and its application.
The present invention is achieved by the following technical solutions：A kind of SSR molecular marker for identifying Gala apple Progeny plants
III, uses simultaneously in detection process, and the molecular labeling is 4 pairs of SSR primers with nucleotide sequence as follows：
LG4 marks CH04e02：
Upstream：5,- GGCGATGACTACCAGGAAAA -3,
Downstream：5,- ATGTAGCCAAGCCAGCGTAT -3,
LG6 marks C53：
Upstream：5,- GCCACTGTGGGTTGCTTTAT -3,
Downstream：5,- AAAACATGCTGCTGTTGGAA -3,
LG12 marks CH01d09：
Upstream：5,- GCCATCTGAACAGAATGTGC -3,
Downstream：5,- CCCTTCATTCACATTTCCAG -3,
LG14 marks CH01g05：
Upstream：5,- CATCAGTCTCTTGCACTGGAAA -3,
Downstream：5,- GACAGAGTAAGCTAGGGCTAGGG -3,。
A kind of application of the SSR molecular marker III for identifying Gala apple Progeny plants, comprises the following steps：
（1）Template is expanded by PCR of loud, high-pitched sound apple genome DNA, respectively with above-mentioned SSR molecular marker as primer pair, enters performing PCR
Amplification, reaction system be 15 μ L, wherein containing the μ L of 10 × PCR Buffer 1.5, the μ L of 2.5 mM dNTPs mixture 1.2,
The μ L of 10ng/ μ L Primers F 1.5, the μ L of 10ng/ μ L Primers R 1.5, the μ L of 5U Taq polymerases 0.15,100ng/ μ L
The μ L of DNA profiling 0.75, deionized water complements to 15 μ L；（2）Amplification program is：94 DEG C of s of 2 min of predegeneration 30,94 DEG C of denaturation
30 s, 60 DEG C of annealing 30 s, 72 DEG C of 40 s of extension, 35 circulate, 72 DEG C of 10 min, and 4 DEG C save backup；（3）PCR primer is examined
Survey, with 8% non-denaturing polyacrylamide electrophoresis, each product of PCR added 1/2 non denatured Loading Buffer,
Mix, 150V constant voltage 150min, glacial acetic acid is fixed, AgNO3Dyeing is taken pictures；（4）It is to be measured when being identified for genetic linkage
Apple can amplify the corresponding specific band of above-mentioned 4 pairs of SSR primers, illustrate the inhereditary material contained in apple germplasm to be measured
Positioned at corresponding genetic linkage group, conversely, the inhereditary material that then apple germplasm to be measured contains is in the absence of corresponding genetic linkage group；
（5）When being identified for anther plant, apple to be measured can amplify the corresponding single spy of above-mentioned 4 pairs of SSR primers
Different in nature band, illustrates that apple plants to be measured, for homozygosis material, two band such as occur, then apple plants to be measured are not homozygosis materials
Material；（6）When being identified for cultivar origin, apple to be measured can amplify any one corresponding specificity of above-mentioned 4 pairs of SSR primers
Band, illustrates that apple plants to be measured, from Gala apple, such as amplify without specific band, then apple plants to be measured are not
The offspring's kind that Gala apple is cultivated.
Compared with prior art, the present invention has advantages below：
1st, the present invention at home and abroad reports apple linkage group residing for expert evidence is identified with SSR molecular marker first,
SSR molecular marker chain on No. 4, No. 6, No. 12 and No. 14 chromosomes of apple is also reported simultaneously；
2nd, molecular labeling of the present invention is codominant marker, and quickly, accurately Gala apple genetic linkage group, flower pesticide can be trained
Support plant and loud, high-pitched sound is cultivated offspring's kind and identified；
3rd, these results of study are utilization and the apple homozygosis that next step accelerates Gala apple and Main Agronomic Characters linked gene
Plant genetic breeding provides the support of molecular level；
4th, draw cultivation plant to carry out molecular level checking for Rapid identification loud, high-pitched sound and provide method.
The present invention is by 4 couple chain on No. 4 to Gala apple anther plant, No. 6, No. 12 and No. 14 chromosomes
SSR molecular marker identification, not only can exactly understand Gala apple genotype type and its genetic diversity, horn of plenty with system
Laid the foundation with development apple allele system, also for scientific utilization Gala apple from now on provides theoretical foundation, Er Qiewei
The genotype of Gala apple anther plant is identified, and its homozygosity is proved.The present invention is selected for Variety of Apple
Educate and molecular mark all has positive impetus.Meanwhile, 4 pairs of SSR molecular markers that the present invention is filtered out
Source-verify is carried out from molecular level provide support for Gala apple offspring's kind.
Brief description of the drawings
Fig. 1 is the Capillary Electrophoresis collection of illustrative plates that LG4 marks CH04e02；Fig. 2 is the Capillary Electrophoresis figure that LG6 marks C53
Spectrum；Fig. 3 is the Capillary Electrophoresis collection of illustrative plates that LG12 marks CH01d09；Fig. 4 is the Capillary Electrophoresis figure that LG14 marks CH01g05
Spectrum.
Specific embodiment
Homozygous genotype is respectively provided with particularly significant effect for the research of higher plant Genetic Mechanisms and Breeding Application
（Murovec, waits Haploids and doubled haploids in plant breeding. In:
Abdurakhmonov I (ed) Plant Breeding: 2012: 87-106.）.Apple is the fruit of genome height heterozygosis
Tree seeds, the assembling difficulty of genome can be substantially reduced using haploid genome（Dunwell. Haploids in
flowering plants:Origins and ex-ploitation. Plant Biotechnol J, 2010,8 (4):
377-424.）.Malus herbaceos perennial, the cyclostage is long, and selfing is not affine in addition, causes to be obtained by inbreeding of more generation
The method of homozygous plants is difficult to.Embryoid is produced to obtain homozygous genotype strain using Anther Culture induction, to genotype
The Apple breeding of height heterozygosis and genetic analysis are significant（Germanà. Gametic embryogenesis and
haploid technology as valuable support to plant breeding. Plant Cell Rep,
2011,30 (5): 839-857；Maria. Doubled haploid production in fruit crops. Plant
Cell, Tissue and Organ Culture, 2006,86：131-146.）.Induced by Anther Culture and produce embryoid
Obtain regeneration plant, and to obtain regeneration plant ploidy and source carry out precise Identification, to Innovation Germplasm genetic analysis have
It is significant.To better profit from these germplasm materials, the present invention enters to the plant that Gala apple and its Anther Culture are obtained
Row linkage group and genotype identification, to accelerate utilization of the Gala apple with Main Agronomic Characters linked gene and apple homozygous plants
Genetic breeding provides the support of molecular level.
Gala apple Anther Culture
After early April Gala apple is buddingged, the method for taking mixing to sample chooses the Gala apple bearing tree of robust growth, adopts
Collect well-developed bud, sealed with hermetic bag and be placed in 4 DEG C of refrigerating chambers of refrigerator and carry out Cold pretreatment.After low-temperature treatment, by flower
Flower bud is sterilized in superclean bench with 0.1% sodium hypochlorite, is then taken out flower pesticide from bud with tweezers and is inoculated in embryoid induction
Culture medium, light culture at 25 DEG C treats that embryoid is long to 8 ~ 10mm after 3 ~ 5 months, being transferred on regeneration culture medium carries out plant again
It is raw, and regeneration plant is obtained through squamous subculture, culture of rootage, domestication, indoor transplanting after carrying out squamous subculture.
SSR molecular marker is analyzed
Genome DNA is extracted
Choose Gala apple and loud, high-pitched sound garland medicine cultivates 6 plants of regeneration plant, using CTAB methods（Cao Qiufen etc., 2003）Flower is extracted respectively
The STb gene of medicine.From the apple high density microsatellite genetic map of the structure such as HiDRAS websites and Okada, selection is distributed in apple
Really No. 4, No. 6, No. 12 and No. 14 the 4 of chromosome pairs of SSR primers, regeneration plant linkage group HIDRAS marks are shown in Table 1；Primer by
Raw work bioengineering（Shanghai）Limited company synthesizes.
Table 1：
PCR is expanded
PCR reaction systems cumulative volume is 15 μ L, by wherein containing the μ L of 10 × PCR Buffer 1.5, the μ of 2.5 mM dNTPs 1.2
The μ L of L, 10ng/ μ L Primers F 1.5, the μ L of 10ng/ μ L Primers R 1.5, the μ L of 5U Taq polymerases 0.15,100ng/ μ
The μ L of L DNA profilings 0.75, deionized water complements to 15 μ L；Amplification program is：94 DEG C of s of 2 min of predegeneration 30,94 DEG C of denaturation
30 s, 60 DEG C of annealing 30 s, 72 DEG C of 40 s of extension, 35 circulate, 72 DEG C of 10 min, and 4 DEG C save backup.
PCR primer is detected
With 8% non-denaturing polyacrylamide electrophoresis, each product of PCR is added 1/2 non denatured Loading
Buffer, is mixed, 150V constant voltages 150min or so, and glacial acetic acid is fixed, and AgNO3 dyeing is taken pictures.
The band obtained to 8% non-denaturing polyacrylamide electrophoresis, is screened, to being drawn and its anther plant in loud, high-pitched sound
The middle mark for producing polymorphism is analyzed, and counts.It is chosen at two allele/sites in Gala apple and all amplifies and carrys out bar
Band, and only one of which allele/site amplifies the primer for carrying out band in Gala apple anther plant.Loud, high-pitched sound apple
In primer CH04e02（LG 4）In, amplified production has the specific band of 139 bp and 144 bp；In primer C53（LG 6）In,
Amplified production has the specific band of 170 bp and 220 bp；In primer CH01d09（LG 12）In, amplified production has 140 bp
With the specific band of 180 bp；In primer CH01g05（LG 14）In, amplified production has the specificity of 100 bp and 140 bp
Band；Result shows, filters out 4 pairs of SSR molecular markers being distributed on the 4th, the 6th, the 12nd and the 14th article of chromosome, is drawn in loud, high-pitched sound
Apple and its flower pesticide amplify allele site in cultivating plant.
Regenerate the ploidy analysis of strain：Regeneration plant blade is taken to be shredded with blade in 500 μ L lysates, it is quiet
Filtered in EP pipes after putting 2 min, with the DNA content in BD Accuri C5 flow cytometry analysis blades after PI dyeing.With
Heterozygous diploid " loud, high-pitched sound " the test tube seedling leaf that Shoot Tip Culture is obtained is the normative reference of Ploidy Identification.
Methods of Ploidy Identification is carried out to surviving regeneration strain using flow cytometer.With heterozygous diploid " loud, high-pitched sound " donor
It is control, as a result shows：Gala21- Gala25 regeneration strain is dliploid.
Regenerate the genotype identification of strain：
The extraction of genomic DNA：After extracting blade STb gene using modified CTAB method, through nucleic acid-protein instrument（Bio-Rad）Detection
DNA concentration, then detect DNA concentration and quality through 1% agarose gel electrophoresis.
5 ' end TAM, FAM, HEX fluorescence labelings of primer are obtained to screening, enters performing PCR amplification.PCR reaction systems are 25 μ
L, mixture containing dNTP（10 mM）0.5 μL、10×PCR Buffer 2.5 μL、25 mM MgCl22.0 μ L, rTaq enzymes
（5 U/μL）0.2 μ L, DNA profiling（100 ng/μL）1 μ L, the μ L of deionized water 17.8.PCR response procedures are：The first step
95 DEG C of predegeneration 3min；Second step 94 DEG C of denaturation 30 s, 60 DEG C of annealing 30 s, 72 DEG C of 30 s of extension, 10 circulations；3rd step
95 DEG C of denaturation 30 s, 55 DEG C of annealing 30 s, 72 DEG C of 30 s of extension, 20 circulations；4th 72 DEG C of step fully extends 6 min, 4 DEG C
Preserve.Reaction plate first adds 9.9 μ L deionized formamides and 0. 1 μ L ROX500 or LIZ500 molecular weight internal standards per hole,
Draw again addition 50 pg amplified productions add sample well in.98 DEG C of 5 min of denaturation, rapidly cool down, and are placed in ABI 3730XL
On DNA analysis instrument.Using Gene Mapper software analysis amplified fragments peak types and read corresponding data.
SSR identification loud, high-pitched sound regeneration plant genotype results are shown in Table 2, as a result show, 2 in linkage group SSR marker shows
Heterozygote is two peaks, and regenerates strain and there was only one of peak.Prove, loud, high-pitched sound regeneration strain Gala21-Gala25 is
Homozygous genotype plant.
The SSR of table 2. identifies loud, high-pitched sound regeneration plant genotype
The botany observation of regeneration strain：Botanical character investigation is carried out to regeneration plant test tube seedling.Regeneration plant botany is observed
3 are the results are shown in Table, the result of table 3 display loud, high-pitched sound heterozygous donors plant height is 5.67 cm（n=1）With the average plant height 2.96 of zygoid
cm ± 0.44 cm（n=28）.We also observe that zygoid growing way is weaker relative to loud, high-pitched sound heterozygous donors simultaneously.It is different
The botanical character of dliploid homozygous plants there is also difference.
Table 3：
Result and analysis
Haploid breeding is one of most effectual way of acquisition advantage parent's donor material.The anther wall of flower pesticide is thin heterozygote
Born of the same parents, while it is likely to induce into the regeneration plant of heterozygous diploid, so the liploid plant for obtaining is not necessarily homozygote.
Just can be the heterozygous diploid of Anther Culture regeneration plant and zygoid area by the allele for identifying regeneration plant
Separate.Technology in the past was all once applied to flower pesticide regeneration plant including isoenzyme mark, S-allele and SSR molecular labelings
Homozygosity identification.The present invention also uses SSR authentication methods.The SSR marker selected first（From HIDRAS databases
（http://www.hidras.unimi.it/））Enter performing PCR amplification to all of regeneration plant, sifting out can distinguish regeneration plant
It is the SSR marker of homozygosis.In order to further discriminate between the genotype of regeneration plant, we filter out again be distributed in apple 4, No. 6,
The SSR marker of linkage group on No. 12 and No. 14 chromosomes.This SSR marker can clearly mark the different plant of " loud, high-pitched sound " apple,
For the genotype identification for carrying out " loud, high-pitched sound " apple from now on provides technical indicator.
The allele that parent is contained can be detected in the plant that Anther Culture is come, but the plant that Anther Culture is come
Middle only one of which allele/site amplifies to be come.This shows that plant its genotype that Anther Culture is come is homozygosis, that is, originate
In the monoploid of parent.
Conclusion
The abundant homozygote plant of ploidy can be also obtained by Anther Culture, this is to carry out apple ploidy genetic breeding research from now on
There is provided abundant examination material.
The present invention obtain regeneration strain and SSR identification systems to analyzing and identifying " loud, high-pitched sound " merit gene studies,
Crossbreeding is transferred in field, and phenotype-genotype association analysis is significant.
Next step combination Gala apple genome sequencing result, plants in genome range to parent and its Anther Culture
Strain is more thoroughly analyzed and compared, to parse Gala apple important character（Characteristic）Molecular mechanism.
Sequence table
The ＞ Research Inst. for fruit Tree, Agricultural Academy of Shanxi Prov. of ＜ 110, Agricultural Biotechnology Research Center of Shanxi Province, Shanxi Province's agriculture
Industry academy of sciences agricultural resource and the institute for economic research
A kind of identification Gala apple SSR molecular marker III and its applications of the ＞ of ＜ 120
〈160〉8
〈210〉1
〈211〉20
〈212〉DNA
The ＞ artificial sequences of ＜ 213
〈220〉
The ＞ LG4 of ＜ 223 mark the sense primer of CH04e02
〈400〉1
GGCGATGACTACCAGGAAAA
〈210〉2
〈211〉20
〈212〉DNA
The ＞ artificial sequences of ＜ 213
〈220〉
The ＞ LG4 of ＜ 223 mark the anti-sense primer of CH04e02
〈400〉2
ATGTAGCCAAGCCAGCGTAT
〈210〉3
〈211〉20
〈212〉DNA
The ＞ artificial sequences of ＜ 213
〈220〉
The ＞ LG6 of ＜ 223 mark the sense primer of C53
〈400〉3
GCCACTGTGGGTTGCTTTAT
〈210〉4
〈211〉20
〈212〉DNA
The ＞ artificial sequences of ＜ 213
〈220〉
The ＞ LG6 of ＜ 223 mark the anti-sense primer of C53
〈400〉4
AAAACATGCTGCTGTTGGAA
〈210〉5
〈211〉20
〈212〉DNA
The ＞ artificial sequences of ＜ 213
〈220〉
The ＞ LG12 of ＜ 223 mark the sense primer of CH01d09
〈400〉5
GCCATCTGAACAGAATGTGC
〈210〉6
〈211〉20
〈212〉DNA
The ＞ artificial sequences of ＜ 213
〈220〉
The ＞ LG12 of ＜ 223 mark the anti-sense primer of CH01d09
〈400〉6
CCCTTCATTCACATTTCCAG
〈210〉7
〈211〉22
〈212〉DNA
The ＞ artificial sequences of ＜ 213
〈220〉
The ＞ LG14 of ＜ 223 mark the sense primer of CH01g05
〈400〉7
CATCAGTCTCTTGCACTGGAAA
〈210〉8
〈211〉23
〈212〉DNA
The ＞ artificial sequences of ＜ 213
〈220〉
The ＞ LG14 of ＜ 223 mark the anti-sense primer of CH01g05
〈400〉8
GACAGAGTAAGCTAGGGCTAGGG
Claims (2)
1. it is a kind of identify Gala apple Progeny plants SSR molecular marker III, it is characterized in that in detection process simultaneously use,
The molecular labeling is 4 pairs of SSR primers with nucleotide sequence as follows：
LG4 marks CH04e02：
Upstream：5,- GGCGATGACTACCAGGAAAA -3,
Downstream：5,- ATGTAGCCAAGCCAGCGTAT -3,
LG6 marks C53：
Upstream：5,- GCCACTGTGGGTTGCTTTAT -3,
Downstream：5,- AAAACATGCTGCTGTTGGAA -3,
LG12 marks CH01d09：
Upstream：5,- GCCATCTGAACAGAATGTGC -3,
Downstream：5,- CCCTTCATTCACATTTCCAG -3,
LG14 marks CH01g05：
Upstream：5,- CATCAGTCTCTTGCACTGGAAA -3,
Downstream：5,- GACAGAGTAAGCTAGGGCTAGGG -3,。
2. the application of a kind of SSR molecular marker III for identifying Gala apple Progeny plants according to claim 1, it is special
Levy is to comprise the following steps：
（1）Template is expanded by PCR of apple genome DNA, respectively with above-mentioned SSR molecular marker as primer pair, enters performing PCR expansion
Increase, reaction system be 15 μ L, wherein containing the μ L of 10 × PCR Buffer 1.5, the μ L of 2.5 mM dNTPs mixture 1.2,
The μ L of 10ng/ μ L Primers F 1.5, the μ L of 10ng/ μ L Primers R 1.5, the μ L of 5U Taq polymerases 0.15,100ng/ μ L
The μ L of DNA profiling 0.75, deionized water complements to 15 μ L；
（2）Amplification program is：94 DEG C of 2 min of predegeneration 30 s, 94 DEG C of denaturation 30 s, 60 DEG C of 30 s of annealing, 72 DEG C extend 40
S, 35 circulations, 72 DEG C of 10 min, 4 DEG C save backup；
（3）PCR primer is detected, with 8% non-denaturing polyacrylamide electrophoresis, by the non-change of each product addition 1/2 of PCR
Property Loading Buffer, mix, 150V constant voltage 150min, glacial acetic acid fixes, AgNO3Dyeing is taken pictures；
（4）When being identified for genetic linkage, apple to be measured can amplify the corresponding specific band of above-mentioned 4 pairs of SSR primers, say
The inhereditary material that contains in bright apple germplasm to be measured is located at corresponding genetic linkage group, conversely, then apple germplasm to be measured contains
Inhereditary material is in the absence of corresponding genetic linkage group；
（5）When being identified for anther plant, apple to be measured can amplify corresponding single one of above-mentioned 4 pairs of SSR primers
Bar specific band, illustrates that apple plants to be measured, for homozygosis material, two band such as occur, then apple plants to be measured are not homozygosis
Material；
（6）When being identified for cultivar origin, it is special that apple to be measured can amplify above-mentioned 4 pairs of SSR primers corresponding any one
Property band, illustrate apple plants to be measured from Gala apple, such as amplified without specific band, then apple plants to be measured are not
It is the offspring's kind of Gala apple cultivation.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2016108494032 | 2016-09-26 | ||
| CN201610849403 | 2016-09-26 |
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| Publication Number | Publication Date |
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| CN106755482A true CN106755482A (en) | 2017-05-31 |
| CN106755482B CN106755482B (en) | 2020-07-17 |
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| 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 |
| 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 |
| 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 |
| 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 |
| 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 |
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| 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 |
| 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 |
| 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 |
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Cited By (6)
| 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 |
| CN110499383A (en) * | 2019-09-02 | 2019-11-26 | 中国农业科学院蔬菜花卉研究所 | Identify the molecular labeling of Chinese cabbage and brassicacarinata interspecific hybrid and progeny material A02 and C02 chromosome separation situation |
| CN111560463A (en) * | 2020-06-15 | 2020-08-21 | 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 |
| CN111663002A (en) * | 2020-07-14 | 2020-09-15 | 福建农林大学 | Microsatellite molecular marker for distinguishing genetic background of second chromosome of high-noble variety and dense variety of cutting hand of sugarcane and application of microsatellite molecular marker |
| 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 |
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| 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 |
| 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 |
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- 2017-01-20 CN CN201710040768.5A patent/CN106755480B/en not_active Expired - Fee Related
- 2017-01-20 CN CN201710040750.5A patent/CN106755478B/en not_active Expired - Fee Related
- 2017-01-20 CN CN201710040784.4A patent/CN106755481B/en not_active Expired - Fee Related
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| CN104531844A (en) * | 2014-12-05 | 2015-04-22 | 中国农业科学院郑州果树研究所 | SSR genotype-based fruit tree variety distinguishing and characteristic fingerprint displaying method |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
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| 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 |
| CN110499383A (en) * | 2019-09-02 | 2019-11-26 | 中国农业科学院蔬菜花卉研究所 | Identify the molecular labeling of Chinese cabbage and brassicacarinata interspecific hybrid and progeny material A02 and C02 chromosome separation situation |
| CN111560463A (en) * | 2020-06-15 | 2020-08-21 | 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 |
| 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 |
| CN111663002A (en) * | 2020-07-14 | 2020-09-15 | 福建农林大学 | Microsatellite molecular marker for distinguishing genetic background of second chromosome of high-noble variety and dense variety of cutting hand of sugarcane and application of microsatellite molecular marker |
| 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 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN106755482B (en) | 2020-07-17 |
| CN106755481B (en) | 2020-07-17 |
| CN106755483B (en) | 2020-07-17 |
| CN106755483A (en) | 2017-05-31 |
| CN106755478B (en) | 2020-07-17 |
| CN106755480B (en) | 2020-07-17 |
| CN106755478A (en) | 2017-05-31 |
| CN106755480A (en) | 2017-05-31 |
| CN106755481A (en) | 2017-05-31 |
| CN106755479A (en) | 2017-05-31 |
| CN106755479B (en) | 2020-07-17 |
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