CN105039336A - Molecular marker subjected to co-segregation with pea powdery mildew resistance allele er1-6 and application thereof - Google Patents
Molecular marker subjected to co-segregation with pea powdery mildew resistance allele er1-6 and application thereof Download PDFInfo
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Abstract
The invention provides a molecular marker subjected to co-segregation with a pea powdery mildew resistance allele er1-6. The molecular marker is located on a VI<th> linkage group of a pea genetic map, and the genetic distance between the molecular marker and the allele er1-6 is 0cM. According to the single base difference (at the position of 1121, T-C) between a disease-resistant variety G0001778 containing the resistance allele er1-6 and an er1 candidate gene PsMLO1cDNA sequence of infected variety pea number 6, primers are designed on the two sides of an SNP mutation site, a high resolution melting curve analysis technology is used for developing the molecular marker subjected to co-segregation with the pea powdery mildew resistance allele er1-6, the marker is subjected to group detection of F2 and F3 which are derived by the disease-resistant variety G0001778 and the infected variety pea number 6 in a hybridization mode, and it is verified that the marker is a co-dominance functional marker subjected to co-segregation with the gene er1-6. Afterwards, the effectiveness is verified in pea powdery mildew resistance resource identification through the marker, the functional marker can be used for molecular marker-assisted selection breeding of the pea powdery mildew resistance, and therefore the breeding process is accelerated.
Description
Technical field
The present invention relates to plant pathology, genetic breeding and technical field of molecular biological detection, specifically, relate to a kind of be divided into pea mildew-resistance allelotrope er1-6 from molecule marker and application.
Background technology
The powdery mildew of pea caused by powdery mildew (ErysiphepisiD.C.) is worldwide disease (Nisaretal., 2011; Fondevillaetal., 2012).In China south, all there is the generation of powdery mildew of pea in pea producing region, north.When weather condition are suitable for, the infection rate of susceptible variety, up to 100%, greatly affects pea yield and quality, causes serious financial loss (Peng Huaxian etc., 1991).Control powdery mildew of pea is the most economical, effective and the method for environmental safety is plantation disease-resistant variety.At present, identify a large amount of mildew-resistance pea resources abroad, and in Resistance resource, identify mildew-resistance gene (er1, er2, er3) (Harlandetal., 1948 of 3 independent inheritances; Heringaetal., 1969).So far, except identifying disease-resistant gene er2 and identify er3 in pea resource JI2480 in pea wild species P.fulvum, the research of a large amount of Resistance resource screening and genetic analysis aspect shows, the resistance of the mildew-resistance pea resource of many different geographic origin is by er1 Gene Handling (Tiwarietal., 1997; Ghafooretal., 2012; Liuetal., 2003; Vaidetal., 1997).The resistance of the mildew-resistance pea resource applied in current production controls by er1.Er1 gene has wide spectrum, durable resistance, widespread use (Fondevillaetal., 2007) in the pea breeding of Europe, North America and Australia.Disease-resistant gene er1 and er2 is positioned in the 6th linkage group (LGVI) (Timmermanetal., 1994) and the 3rd linkage group (LG III) (Katochetal., 2010) of pea genetic map respectively.
Nearest research finds, pea mildew-resistance gene er1 produces (Humphryetal., 2011 by susceptible gene PsMLO afunction; Pavanetal., 2011).Under nature and mutagenic condition, pea PsMLO homologous sequence generation base deletion, insertion, replacement etc. cause different er1 allelotrope to produce, 5 er1 allelotrope and er1-1, er1-2, er1-3, er1-4 and er1-5 (Humphryetal., 2011 are identified at present; Pavanetal., 2011).Except er1-2 is produced by the insertion of large fragment and disappearance, other 4 allelotrope are all produced by the disappearance of single base, insertion, replacement.Along with the widespread use of molecular marking technique in marker assisted selection, Pavan etc. (2013) are based on different molecular marking techniques, develop 5 allelic functional labels of the er1 identified, but the validity of these functional labels is not yet verified.High resolving power melting curve (HighResolutionMelting, HRM) analytical technology is the one detection gene mononucleotide polymorphism (SingleNucleotidePolymorphism of rising in recent years, SNP) new technology (Kristensenetal., 2008).
Nearest researchist has filtered out some Powdery Mildews immunity resource (Wang Zhongyi etc., 2012) in Chinese Local Varieties of Pea.In Chinese Local Varieties of Pea, a neomorph er1-6 is identified by the candidate gene PsMLO sequential analysis of er1, this gene is undergone mutation at PsMLO1cDNA sequence open reading frame the 1121st base place, T replaces with C, this base mutation causes aminoacid sequence to become proline(Pro) by leucine, thus causes the change of protein function.For acceleration molecular assisted selection, exploitation with allelotrope er1-6 be divided into from functional molecular marker become technical problem urgently to be resolved hurrily.
Summary of the invention
The object of this invention is to provide a kind of be divided into pea mildew-resistance allelotrope er1-6 from molecule marker and application.
In order to realize the object of the invention, of the present invention a kind of be divided into pea mildew-resistance allelotrope er1-6 from molecule marker, described molecule marker is positioned in pea genetic map VI linkage group, be 0cM with the genetic distance of allelotrope er1-6, for the Auele Specific Primer of molecule marker described in pcr amplification to as follows, amplified production size is 158bp;
Upstream primer F:5 '-CTGGAGATCACCTTTTCTGGTT-3 ' and
Downstream primer R:5 '-CATGTACAAACACACATACACACG-3 '.
Wherein, the annealing temperature that pcr amplification uses is 59 DEG C.
The present invention also provides the described application of molecule marker in qualification pea mildew-resistance germ plasm resource.
Particularly, described application comprises the following steps:
1) genomic dna of pea to be measured is extracted;
2) with the genomic dna of pea to be measured for template, the Auele Specific Primer pair for molecule marker described in pcr amplification described in utilization, carries out pcr amplification reaction;
3) HRM analytical technology is utilized to detect pcr amplification product.
Wherein, the amplification system that pcr amplification reaction uses is counted with 10 μ l: pea genomic dna 25ng, 10 × PCR reaction buffer (25mMMgCl
2) 1 μ l, 5mM primers F 0.25 μ l, 5mM primer R0.25 μ l, 10 × HRMFastMasterMix (HRMFastPCRKit, KapaBiosystems, USA) 5 μ l, ddH
2o complements to 10 μ l.
The condition of pcr amplification reaction is: 95 DEG C 5 minutes; 94 DEG C 10 seconds, 59 DEG C 30 seconds, 72 DEG C 10 seconds, 50 circulations; 72 DEG C 10 minutes.
The present invention utilizes HRM analytical technology to detect pcr amplification product, and an identical melting curve all appears in all pea resources containing allelotrope er1-6, and other disease-resistant or susceptible pea resources all form other one and the visibly different melting curve of er1-6.
The present invention is also provided for the test kit that PCR detects mildew-resistance pea resource, containing the described Auele Specific Primer pair for molecule marker described in pcr amplification in described test kit.
Preferably, described test kit also comprises dNTPs, Taq DNA polymerase, Mg
2+, PCR reaction buffer, HRMFastMasterMix, at least one in standard positive template etc.
The present invention further provides the application of described molecule marker in plant molecular marker assistant breeding.
The present invention also provides the test kit for detecting mildew-resistance pea containing above-mentioned primer pair.Preferred described test kit also comprise in above-mentioned MasterMix (Tiangen) and HRMFastMasterMix (HRMFastPCRKit, KapaBiosystems, USA) and standard positive template one or more.
Particularly, of the present invention be divided into pea mildew-resistance er1 allelotrope er1-6 from molecule marker obtain by the following method:
Utilize the SNP difference (T → C) of the PsMLO1cDNA sequence of disease-resistant variety G0001778 and No. 6, susceptible variety dam pea, primer is designed respectively in both sides, PsMLO1cDNA sequence SNP mutational site, exploitation with pea mildew-resistance allelotrope er1-6 be divided into from molecule marker, subsequently, this mark is hybridized derivative F for the identification of disease-resistant parent G0001778 and No. 6, Susceptible parent dam pea
2the genotype of each individual plant of colony, by HRM technical Analysis, this is marked at F
2there are three kinds of visibly different melting curves in colony, correspond respectively to the disease-resistant gene type that isozygotys, susceptible genotype of isozygotying and heterozygous genotypes three types.This three kinds of genotype and F
3the phenotype one_to_one corresponding of colony.Prove this be labeled as to be divided into disease-resistant gene er1-6 from molecule marker.Subsequently, by described molecule marker for the identification of pea mildew-resistance resource.Result shows, described molecule marker effectively can distinguish the pea resource containing mildew-resistance allelotrope er1-6.The present invention provides molecule marker for pea mildew-resistance molecule assisted selection, thus accelerates pea mildew-resistance breeding work process.
Specific embodiments:
(1) the Functional marker exploitation of er1-6
According to the candidate gene PsMLO1 sequence alignment result of the disease-resistant variety G0001778 containing er1-6 and No. 6, susceptible variety dam pea, there are single base difference (1121 places between er1-6 and PsMLO1cDNA sequence, T → C), with PrimerPremier5.0 software at both sides, sequence SNP mutational site design primer, development functionality marks.On the 11st exon that the upstream and downstream primer of this mark lays respectively at PsMLO1 gene and the 11st intron.Primer is synthesized by Sangon Biotech (Shanghai) Co., Ltd..
First, be that template carries out pcr amplification with G0001778 and dam pea No. 6 genomic dnas, detect the validity of primer, utilize the polymorphism between HRM analytical technology detection parent subsequently.Analyzed by HRM, find that the PCR primer that a molecule marker increases in anti-, sense kind defines two kinds of visibly different melting curves, illustrate that this is marked between disease-resistant variety G0001778 and No. 6, susceptible variety dam pea and has polymorphism.Subsequently the molecule marker between the parent of exploitation with polymorphism is hybridized derivative F for the identification of disease-resistant parent G0001778 and No. 6, Susceptible parent dam pea
2the genotype of 71 individual plants of colony.Find that this is marked at F
2form visibly different three kinds of melting curves in colony, correspond respectively to disease-resistant, susceptible and heterozygosis three kinds of genotype of isozygotying of isozygotying, these three kinds different curves and F
3the Characters type one_to_one corresponding (Fig. 1) of colony, this show described molecule marker be divided into er1-6 from codominance functional label.
(2) application of Functional marker
The molecule marker of exploitation is carried out applying and functional verification in the qualification of pea mildew-resistance resource.Result shows, in the pea resource of all detections, all can amplify the object band of a 158bp.PCR primer is through HRM technical Analysis, and an identical melting curve (Fig. 3), all appears in all pea resources containing mildew-resistance allelotrope er1-6.And containing the pea resource of other er1 allelotrope er1-1, er1-2, er1-4 and susceptible resource all formed other one with the visibly different melting curve of er1-6 (Fig. 3).Analytical results proves that described molecule marker accurately can distinguish the pea resource containing disease-resistant allelotrope er1-6.The validity of the described molecule marker of this result verification in the qualification of pea mildew-resistance resource.
This invention exploits to be divided into pea mildew-resistance allelotrope er1-6 from molecule marker, this disease-resistant gene er1-6 identifies in China's pea resource, to China's powdery mildew of pea, there is resistance of wide spectrum, effectively can control the generation of China's powdery mildew of pea, alleviate the production loss that this disease causes.PCR-based amplification and HRM analytical technology, by with this gene be divided into from molecule marker be effectively applied in molecular mark, overcome the shortcoming that cycle required for conventional breeding methods grows; HRM has high-throughput, advantage easy and simple to handle; The present invention utilizes that the molecule marker of HRM technological development is produced at pea, have important value in the research of breeding work and anti-disease mechanism.Its advantage is:
(1) of the present invention be divided into pea mildew-resistance allelotrope er1-6 from molecule marker, be obtain and the functional label verified in the filial generation family of G0001778 and resistance thereof.Effectively can be applied to the molecular mark of the qualification of pea mildew-resistance resource and pea mildew-resistance progeny population.
(2) utilize of the present invention be divided into pea mildew-resistance allelotrope er1-6 from molecule marker, can successful identification containing the pea resource of mildew-resistance gene er1-6, thus prove that this molecule marker has validity in the qualification of pea mildew-resistance resource.
Embodiment
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.If do not specialize, embodiment is experiment condition all conveniently, as Sambrook equimolecular Cloning: A Laboratory Manual (SambrookJ & RussellDW, Molecularcloning:alaboratorymanual, 2001) condition of, or according to manufacturer's specification sheets advising.
The er1 candidate gene PsMLO1cDNA Sequence Identification of embodiment 1 Resistance resource and the qualification of neomorph er1-6
The total serum IgE of 15 pea Resistant germplasm and No. 6, susceptible check variety dam pea is extracted with RNAprep plant total RNA extraction reagent box (centrifugal column type, purchased from Tiangen company).Concrete operation method is as follows:
1) get about 100mg pea young leaflet tablet rapid grind into powder in liquid nitrogen, add 450 μ LRL (adding beta-mercaptoethanol to final concentration before use is 1%), vortex concuss mixes; 2) all solution is transferred to (Filter column CS is placed in collection tube) on Filter column CS, the centrifugal 5min of 12000rpm, the supernatant in careful absorption collection tube is in the centrifuge tube of RNase-free; 3) in centrifuge tube, slowly add the dehydrated alcohol of 0.5 times of volume, mixing, the solution obtained is proceeded in adsorption column CR3 together with precipitation, leave standstill the centrifugal 1min of 2min, 12000rpm, outwell the waste liquid in collection tube; 4) in adsorption column CR3, add 500 μ L protein liquid removal RW1, leave standstill the centrifugal 1min of 2min, 12000rpm, outwell the waste liquid in collection tube; 5) preparation of DNase I working fluid: get 10 μ LDNase I storage liquid and put into new RNase-free centrifuge tube, add 70 μ LRDD solution, softly mix; 6) add DNase I working fluid of 80 μ L to adsorption column CR3 central authorities, room temperature places 20min; 7) in adsorption column CR3, add 350 μ L protein liquid removal RW1, leave standstill the centrifugal 1min of 2min, 12000rpm, outwell the waste liquid in collection tube; 8) in adsorption column CR3, add 500 μ L rinsing liquids RW (adding dehydrated alcohol before use), room temperature leaves standstill 2min, and the centrifugal 1min of 12000rpm, outwells the waste liquid in collection tube; 9) repeating step 8; 10) the centrifugal 2min of 12000rpm, outwells waste liquid, adsorption column CR3 is placed in room temperature and places several minutes, thoroughly dries residual rinsing liquid; 11) adsorption column CR3 is put into a new RNase-free centrifuge tube, the unsettled dropping 50 in the mid-way to adsorption film μ LRNase-freeddH
2o, room temperature places the centrifugal 2min of 10min, 12000rpm, obtains RNA solution; 12) RNA integrity detection: the gel electrophoresis of plain agar sugar, deposition condition: gum concentration 1.2%; 1 × TBE electrophoretic buffer; 150V; 15min.
MRNA the 1st article of cDNA chain is synthesized with BioRT reverse transcription amplification (RT-PCR) test kit (two-step approach), with PsMLO1 Auele Specific Primer, pcr amplification (Pavenetal., 2013) is carried out to PsMLO1F/PsMLO1R (5 '-AAAATGGCTGAAGAGGGAGTT-3 '/5 '-TCCACAAATCAAGCTGCTACC-3 ').Pcr amplification product detects through 1.5%TBE agarose gel electrophoresis, adopts plain agar sugar gel DNA to reclaim test kit (centrifugal column type, Tiangen) and reclaims and purified pcr product.PCR primer after purifying is cloned in pZeroBack carrier or pMG-T carrier (Tiangen), deliver to Beijing Liuhe Huada Genomics Technology Co., Ltd's order-checking, utilize ClustalX2 software analysis anti-sense parent between PsMLO1 sequence difference, and with known PsMLO1 sequence (FJ463618) compare of analysis (Humphryetal.2011).
The qualification of neomorph er1-6:
PsMLO1cDNA sequence alignment analysis result shows that the PsMLO1cDNA sequence of No. 6, susceptible contrast dam pea is identical with wild-type PsMLO1 sequence (FJ463618).This demonstrate that the accuracy of the disease-resistant gene sequence of gained.Disease-resistant gene PsMLO1 contained by 15 Resistance resource has identical cDNA sequence, and this sequence, at 1121 places of wild-type PsMLO1cDNA sequence (FJ463618), single base mutation (T → C) (SEQIDNO.5) occurs.This base mutation causes aminoacid sequence to become proline(Pro) by leucine, thus causes the change of protein function.This is all not identical with 5 the allelic mutated site of er1 identified, shows that this is a neomorph of er1, by its called after er1-6 (table 1).
Table 1 resistance Local Varieties of Pea is to the phenotypic response of powdery mildew EPYN and contained er1 allelotrope
*: 0,1,4 represent Resistant reaction grade respectively.
Embodiment 2 and pea mildew-resistance allelotrope er1-6 be divided into from the exploitation of molecule marker
According to gene PsMLO1cDNA sequence and the wild-type PsMLO1 sequence alignment result of pea mildew-resistance kind G0001778 and No. 6, susceptible variety dam pea, discovery G0001778 contains a new allelotrope er1-6, this allelotrope PsMLO1 sequence open reading frame the 1121st place occur single base replace (T → C), with PrimerPremier5.0 software at this design upstream and downstream, both sides, SNP mutational site primer development functional molecular marker.
Upstream primer F:5 '-CTGGAGATCACCTTTTCTGGTT-3 '
Downstream primer R:5 '-CATGTACAAACACACATACACACG-3 '
On the 11st exon that the upstream and downstream primer of this functional label lays respectively at PsMLO1 gene and the 11st intron.Primer is synthesized by Sangon Biotech (Shanghai) Co., Ltd..
First be that template carries out pcr amplification with G0001778 and dam pea No. 6 genomic dnas, detect the validity of primer.
The amplification system that pcr amplification reaction uses is counted with 10 μ l: pea genomic dna 25ng, containing 25mMMgCl
210 × PCR reaction buffer 1 μ l, 5mM primers F 0.25 μ l, 5mM primer R0.25 μ l, 10 × HRMFastMasterMix5 μ l, ddH
2o complements to 10 μ l.
The condition of pcr amplification reaction is: 95 DEG C 5 minutes; 94 DEG C 10 seconds, 59 DEG C 30 seconds, 72 DEG C 10 seconds, 50 circulations; 72 DEG C 10 minutes.
Result shows, disease-resistant variety G0001778 and No. 6, susceptible variety dam pea all amplify the object fragment of a 158bp.Subsequently, utilize HRM technology for detection PCR primer in polymorphism that is anti-, sense kind.HRM analytical results display molecule marker, resisting, feeling formation two visibly different melting curves between kind, represents two kinds of genotype respectively.Show that molecule marker has polymorphism between anti-, sense kind.Subsequently this molecule marker is carried out F
2colony's checking, is marked at G0001778 and No. 6, dam pea hybridizes derivative F
2all amplify the PCR primer of a 158bp in colony, by HRM technical Analysis, occur three kinds of visibly different melting curves in colony, these three kinds of curve types correspond respectively to disease-resistant, susceptible and heterozygosis three kinds of genotype (Fig. 1) of isozygotying of isozygotying.And be marked at F
2clastotype in colony passes through X
2inspection, meets 1:2:1 ratio, shows that described molecule marker is for codominant marker.Three kinds of melting curves respectively with F
3isozygotying of colony be disease-resistant, isozygoty the susceptible and anti-phenotype one_to_one corresponding feeling the strain be separated, this show this molecule marker be divided into disease-resistant gene er1-6 from functional label.By mapmaker3.0 computed in software genetic linkage distance, MapDraw is used to build genetic linkage maps.The genetic distance of this mark and disease-resistant gene er1-6 is 0cM, described molecule marker be divided into er1-6 from functional label (Fig. 2).
Embodiment 3 and pea mildew-resistance allelotrope er1-6 be divided into from the application of molecule marker
The functional molecular marker of the er1-6 of exploitation is carried out in Foreign Banks' Entries resource apply and functional verification.Result shows, described molecule marker all amplifies the object band of a 158bp in the disease-resistant of all detections and susceptible pea resource.By HRM analytical technology, pea resource (G0001752, G0001763, G0001764, G0001768, G0001778 containing disease-resistant allelotrope er1-6, through PsMLO1cDNA Sequence Identification, all there is single base mutation T → C at the 1121bp place of PsMLO1cDNA sequence) form identical melting curve (Fig. 3).Containing other er1 allelotrope er1-1 (Tara), er1-2 (X9002 and G0005576), the Resistant germplasm of er1-4 (YI), and susceptible resource (No. 6, dam pea, No. 1, Gansu Province pea, G0001747, G0003839, G0003840) all forms one and the visibly different melting curve of er1-6 (Fig. 3).Result shows that this mark utilizes HRM analytical technology can go out pea resource containing disease-resistant gene er1-6 by precise Identification.The validity of the described molecule marker of this result verification in the qualification of pea mildew-resistance resource.
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.
Reference
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Accompanying drawing explanation
Fig. 1 be utilize in the embodiment of the present invention 2 HRM technical Analysis and pea mildew-resistance allelotrope er1-6 be divided into from molecule marker hybridize derivative F at disease-resistant variety G0001778 and No. 6, susceptible variety dam pea
2the melting curve figure of part individual plant in colony; Wherein, 1 is the disease-resistant gene type CC that isozygotys, and 2 is heterozygous genotypes CT, and 3 is the susceptible genotype TT that isozygotys.
Fig. 2 is shown as in the embodiment of the present invention 2 based on the F that disease-resistant variety G0001778 and the hybridization of No. 6, susceptible variety dam pea derive
2the pea genetic map VI linkage group of informative population, this genetic map contain that the molecule marker chain with er1-6 and the present invention develop with er1-6 be divided into from Functional marker, genetic distance unit is cM.
Fig. 3 be utilize in the embodiment of the present invention 3 HRM technical Analysis and pea mildew-resistance allelotrope er1-6 be divided into from the melting curve figure that formed in pea mildew-resistance resource is identified of molecule marker; Wherein, 4 is the genotype of er1-6, and 5 is the genotype of non-er1-6.
Claims (10)
1. with pea mildew-resistance allelotrope er1-6 be divided into from molecule marker, it is characterized in that, described molecule marker is positioned in pea genetic map VI linkage group, be 0cM with the genetic distance of allelotrope er1-6, for the Auele Specific Primer of molecule marker described in pcr amplification to as follows, amplified production size is 158bp;
Upstream primer F:5 '-CTGGAGATCACCTTTTCTGGTT-3 ' and
Downstream primer R:5 '-CATGTACAAACACACATACACACG-3 '.
2. molecule marker according to claim 1, is characterized in that, the annealing temperature that pcr amplification uses is 59 DEG C.
3. the application of the molecule marker described in claim 1 or 2 in qualification pea mildew-resistance germ plasm resource.
4. application according to claim 3, is characterized in that, described application comprises the following steps:
1) genomic dna of pea to be measured is extracted;
2) with the genomic dna of pea to be measured for template, utilize the Auele Specific Primer pair for molecule marker described in pcr amplification described in claim 2, carry out pcr amplification reaction;
3) HRM analytical technology is utilized to detect pcr amplification product.
5. application according to claim 4, is characterized in that, step 2) in pcr amplification reaction use amplification system count with 10 μ l: pea genomic dna 25ng, containing 25mMMgCl
210 × PCR reaction buffer 1 μ l, 5mM primers F 0.25 μ l, 5mM primer R0.25 μ l, 10 × HRMFastMasterMix5 μ l, ddH
2o complements to 10 μ l.
6. application according to claim 4, is characterized in that, step 2) in carry out pcr amplification reaction condition be: 95 DEG C 5 minutes; 94 DEG C 10 seconds, 59 DEG C 30 seconds, 72 DEG C 10 seconds, 50 circulations; 72 DEG C 10 minutes.
7. the application according to any one of claim 4-6, it is characterized in that, step 3) in utilize HRM analytical technology detect pcr amplification product, all there is an identical melting curve in all pea resources containing allelotrope er1-6, and other disease-resistant or susceptible pea resources all form other one and the visibly different melting curve of er1-6.
8. detect the test kit of mildew-resistance pea resource for PCR, it is characterized in that, containing the Auele Specific Primer pair for molecule marker described in pcr amplification described in claim 1 or 2 in described test kit.
9. test kit according to claim 8, is characterized in that, described test kit also comprises dNTPs, Taq DNA polymerase, Mg
2+, PCR reaction buffer, HRMFastMasterMix, at least one in standard positive template.
10. the application of the molecule marker described in claim 1 or 2 in plant molecular marker assistant breeding.
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| CN201510542741.7A Pending CN105039336A (en) | 2015-08-28 | 2015-08-28 | Molecular marker subjected to co-segregation with pea powdery mildew resistance allele er1-6 and application thereof |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107058494A (en) * | 2017-01-13 | 2017-08-18 | 兰州大学 | Simplify the method for Jian Kuo Foreign Banks' Entries Purities using SCoT molecular labelings |
| CN108546777A (en) * | 2018-07-19 | 2018-09-18 | 上海市农业科学院 | A kind of SNP marker and its application for detecting the anti-clubroot of Chinese cabbage |
| CN111826454A (en) * | 2019-04-23 | 2020-10-27 | 江苏省农业科学院 | Molecular marker VrMLO _ Indel2 for identifying powdery mildew resistance phenotype of mung beans as well as primer and application thereof |
| WO2023207233A1 (en) * | 2022-04-25 | 2023-11-02 | 山东省农业科学院 | Neutral snapshot marker of peas and use thereof in population genetic diversity analysis |
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| 付海宁: "豌豆抗白粉病表型及基因型鉴定", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107058494A (en) * | 2017-01-13 | 2017-08-18 | 兰州大学 | Simplify the method for Jian Kuo Foreign Banks' Entries Purities using SCoT molecular labelings |
| CN107058494B (en) * | 2017-01-13 | 2020-09-01 | 兰州大学 | Method for simplifying purity identification of common vetch variety by adopting SCoT molecular marker |
| CN108546777A (en) * | 2018-07-19 | 2018-09-18 | 上海市农业科学院 | A kind of SNP marker and its application for detecting the anti-clubroot of Chinese cabbage |
| CN111826454A (en) * | 2019-04-23 | 2020-10-27 | 江苏省农业科学院 | Molecular marker VrMLO _ Indel2 for identifying powdery mildew resistance phenotype of mung beans as well as primer and application thereof |
| CN111826454B (en) * | 2019-04-23 | 2022-08-02 | 江苏省农业科学院 | Molecular marker VrMLO _ Indel2 for identifying powdery mildew resistance phenotype of mung beans as well as primer and application thereof |
| WO2023207233A1 (en) * | 2022-04-25 | 2023-11-02 | 山东省农业科学院 | Neutral snapshot marker of peas and use thereof in population genetic diversity analysis |
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