CN105543366A - Development and application of internal specificity-SNP codominant molecular markers of rice blast-resistance gene Pi25 gene - Google Patents
Development and application of internal specificity-SNP codominant molecular markers of rice blast-resistance gene Pi25 gene Download PDFInfo
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Abstract
The invention discloses internal specificity-SNP codominant molecular marker primers of a rice blast-resistance gene Pi25 gene and an application. An applicant obtains an internal specificity-SNP locus of the Pi25 gene by carrying out sequencing, sequence alignment and identification on different allelic genes and develops the set of molecular marker primers for identifying the SNP, the molecular marker primers include Pi25-PF:GGGATCTAGGCAGACAGT, Pi25-PR:CAGAGCAGGAACTTCAGTTG, Pi25-NR:GATGCAGGATAAAGAATGAA, Pi25-NF:AGCTGTGGCAAGCCTAACAG. Rice DNA is subjected to PCR amplification through the primers, and the genotype of to-be-detected rice Pi25 can be rapidly judged. A method is easy, convenient, rapid and low in cost, and can be widely applied to molecular-marker-assisted selection breeding or rice-germplasm-resource Pi25 genotype identification.
Description
Technical field
The invention belongs to molecular genetics field, be specifically related to the exploitation of special SNP codominant marker and application in rice blast resistant gene Pi 25 for rice gene, molecule marker primer provided by the invention to be suitable in rice breeding the genotypic a large amount of screening of Pi25 and from Rice Germplasm Resources, to screen new Pi25 genotypic resistance parent.
Technical background
Paddy rice is above the Major Foods of 50% world population.Along with the continuous increase of world population, the demand of rice yield is increased day by day.Rice blast is one of disease that China's rice workspace is the most serious, is also the threat of whole world Rice Production.It is reported that the paddy rice that the whole world is caused by rice blast between 1975-1990 loses up to 1.57 × 1011kg, China's rice blast year onset area about 3,800,000 hectares, onset area accounts for 6%, year loss paddy about 1,000,000,000 kilograms, since 2000 in southwest, northeast is in laying particular stress on epidemic status, because hazard area and hazard rating are on the rise, rice blast has become the serious hindrance of rice high yield stable yields.Selection and popularization disease resisting rice kind (combination) be control that rice blast disease is the safest, the method for economy and environmental protection, the excavation of disease-resistant gene, evaluation and rationally application become the basis of current paddy disease-resistant breed and production.But in actual production, rice blast Race is very fast, resistant variety life-time service containing single resistant gene just can because of the appearance of the new dominant population of Pyricularia oryzae and resistance is gradually lost, therefore, and utilize molecular marker assisted selection to assemble, polymerization has the resistant gene of different anti-spectrum, cultivate wide spectrum, durable resistance rice varieties become the optimal path of dealing with problems.
Pi25 is positioned at paddy rice No. 6 the short arm of a chromosome near centric region (ChenJieetal.APid3allelefromricecultivarGumei2confersresi stancetoMagnaportheoryzae.JournalofGeneticsandGenomics38 (2011) 209-216), its donor parents derives from Gumei2, be one and there is kind rice blast physiological strain to resistance of wide spectrum therefore for cultivating the rice varieties with resistance of wide spectrum, there is huge utility value.Current allelomorphism method for measuring is by carrying out different rice blast physiological strain inoculated identification to containing not homoallelic material in greenhouse, to isoallele not make a distinction according to their anti-spectral difference is different, but it is long, complicated, loaded down with trivial details that this method exists experimental period, because which limit its application in breeding.
Molecule marker is a kind of quick, easy, with low cost and can carry out a kind of technology of nondestructive testing in early days at paddy growth, but at present also not to the report of Pi25 gene internal specific codominant marker.This research has carried out pcr amplification by the allelotrope of the Pi25 coding region to Pi25 source material, allelotrope is resurveyed sequence, finally after Pi25 Gene A TG promotor ,+2566bp locates acquisition Pi25 specificity SNP site, Pi25 allelotype is C base, all the other genotype are G base, at two couples of cross primer PCR (PCRwithconfrontingtwo-pairprimers, reformed AHP in addition on the basis of PCR-CTPP) method, by introducing base mismatch in inner primer antepenulatimate, develop the codominant marker of this SNP site Different Alkali base type of qualification, as shown in Figure 1, by designing 4 primer (PF, PR, NF, and utilize variety genome DNA to be measured to carry out pcr amplification NR), according to amplified band type identification Pi25 genotype, if amplify 383bp band, it is Pi25 genotype of isozygotying, all the other allelic gene type amplified bands are 266bp, heterozygous then has two band simultaneously, and three also has the total band of a 612bp.Therefore the method is easy to be quick, with low cost, can be widely used in molecular marker assisted selection breeding or Rice Germplasm Resources Pi25 genotype identification.
Summary of the invention
The object of the present invention is to provide special SNP codominant marker primer in rice blast resistant gene Pi 25 for rice gene, be respectively: Pi25-PF:GGGATCTAGGCAGACAGT; Pi25-PR:CAGAGCAGGAACTTCAGTTG; Pi25-NR:GATGCAGGATAAAGAATGAA; Pi25-NF:AGCTGTGGCAAGCCTAACAG.Primer specificity is good, and amplification efficiency is high, can be used for the genotype identification of rice blast resistant gene Pi 25 for rice.
Another object of the present invention there is provided the application of rice blast resistant gene Pi 25 for rice special SNP codominant marker primer, utilize this primer effectively can carry out genotype selection to blast resistant gene Pi 25 for rice, both may be used for the Screening and Identification of rice pest insects, may be used for again the molecular breeding of rice blast resistant gene Pi 25 for rice.
To achieve these goals, this invention takes following technical measures:
Technical solution of the present invention takes following thinking: Pi25 is positioned at paddy rice No. 6 chromosomal galianconism near centric region, ORF total length 2775bp, intronless.The allelotrope of applicant to the Pi25 coding region of Pi25 source material has carried out pcr amplification, allelotrope resurveys sequence, by acquisition sequence first with comprise fine and 9311 the corresponding sequence of susceptible allelic material Japan and compare, all the other 3 allelic materials are comprised for the site recycling with diversity sequence and carries out order-checking comparison confirmation, finally after Pi25 Gene A TG promotor ,+2566bp locates acquisition Pi25 specificity SNP site, Pi25 allelotype is C base, and all the other genotype are G base; Therefore, the allelic qualification of Pi25 can be realized by design Auele Specific Primer to this SNP site specific amplification.
Design of primers principle following (Fig. 1): when designing C type Identity of allele primer, according to PCR-CTPP Method And Principle, first in this SNP site upstream design forward primer PF (table 1), using the complementary base G of the C base of this SNP site as 3 ' end at this site design reverse primer PR, 3 ' of the PR G base of holding and C type allelotrope material are normally combined and increase and form G/G mispairing with G shaped material and cannot increase in theory, and our experimental result shows that this mispairing can not distinguish two kinds of genotype completely, also fainter amplification is had in G shaped material, therefore in order to strengthen this primer specificity, we introduce a base mismatch T in the antepenulatimate of PR and strengthen its specificity, result shows that this primer only has when increasing with C type allelotrope material and has a 383bp band, band is not had to increase with G shaped material, in like manner, NF and the NR of amplification G type allelotrope material is developed according to above-mentioned thinking, article 4, Primer, sequence, Tm value and expanding fragment length are shown in Table 1.
In rice blast resistant gene Pi 25 for rice gene, special SNP codominant marker primer, is respectively: Pi25-PF:GGGATCTAGGCAGACAGT; Pi25-PR:CAGAGCAGGAACTTCAGTTG; Pi25-NR:GATGCAGGATAAAGAATGAA; Pi25-NF:AGCTGTGGCAAGCCTAACAG.
The application of special SNP codominant marker primer in rice blast resistant gene Pi 25 for rice gene, comprise and utilize usual manner to carry out pcr amplification to paddy DNA, by carrying out genotypic judgement to amplified band, for the Screening and Identification of rice pest insects or the molecular breeding of rice blast resistant gene Pi 25 for rice.
In above-described scheme, preferably, primer PF, PR, NF and NR press 1:1:1.5:1.5 mixing amplification, and annealing temperature is 56 DEG C.
In above-described scheme, the judgment mode of amplified band is: SNP site is that the Pi25 type allelotrope material of C base amplifies 612bp and 383bp2 band, SNP site is that its alloytype allelotrope material of G base amplifies 612bp and 266bp2 band, and GC heterozygous then amplifies 3 band of 612bp, 383bp and 266bp.
Table 1 primer sequence
Compared with prior art, the present invention has the following advantages:
1. applicant design the Pi25 coding region of primer pair Pi25 source material allelotrope has carried out pcr amplification, allelotrope resurveys sequence, finally after Pi25 Gene A TG promotor ,+2566bp locates acquisition Pi25 specificity SNP site, Pi25 allelotype is C base, and all the other genotype are G base.
2. inoculate different rice blast Race Identification rice blast resistance allelotrope from traditional utilization different, the present invention is improved on the basis of PCR-CTPP method, utilizes SNP site in target gene to develop and only utilizes PCR electrophoretic technique can identify parent and F
2the genotypic special codominant marker of filial generation Pi25.
3. when utilizing Pi25 gene pairs Rice Resistance To Rice Blast to improve, molecular marker assisted selection (MAS) becomes the most direct, the effective screening implement of one in backcross process, utilize method of the present invention accurately can realize progeny genotypes to detect, after the individual continuous backcross of goal gene is carried in screening, selfing is isozygotied thus completes breed improvement; Therefore, the method has simply, accurately, cost is low and can carry out the series of advantages such as non-damaged data in early days at paddy growth, is suitable in breeding to the genotypic a large amount of screening of Pi25 and for Pi25 allelotype qualification in Rice Germplasm Resources.
Accompanying drawing explanation
Fig. 1 is for utilizing two pairs of cross primer pcr amplification (PCR-CTPP) methods to SNP site amplification schematic diagram special in paddy rice Pi25 gene.
Fig. 2 is for utilizing 3 different annealing temperature (54 DEG C, 56 DEG C, 58 DEG C), 3 kinds of different primers concentration ratios (PF, PR, NF and NR concentration ratio are respectively 1:1:1:1,1:1:1.5:1.5 and 1:1:2:2) to the electrophorogram of 3 kinds of different Pi25 genotype DNA profiling pcr amplifications
M:DL2000DNAmarker; Wherein AA section mould plate and swimming lane 1,4,7 are for SNP site is for isozygotying C base, the parent material Gumei2 containing Pi25 gene; GG section mould plate and swimming lane 3,6,9 for SNP site is for isozygotying G base, the susceptible allelotrope parent material 9311 containing Pi25; CG section mould plate and swimming lane 2,5,8 are CG heterozygous base for SNP site, hybridize by Gumei2 and 9311 F obtained
1for material.
Fig. 3 is that the Pi25 molecule marker primer utilizing the present invention to develop detects the electrophorogram producing upper conventional backbone parent.
M:DL2000DNAmarker; P1, P2 are parent's Gumei2 that target SNP site is respectively C and G base
(Pi25) with 9311 (susceptible allelotrope), F
1for Gumei2 and 9311 first-filial generation, swimming lane 1-20 is respectively and produces upper conventional backbone parent: Mianhui725, bright extensive 63, Chinese Fragrant Rice, Japan are fine, osmanthus towards No. 2, Huang Huazhan, Lemont, B5, HP121, more light, the super Bath agate base of a fruit, salt rice No. 4, R1128, Peiai 64S, Guangzhan 63S, HD9802S, II-32A, golden 23A, the rich A in sky, huge wind 2A, belonging to each material, genotype marks below corresponding swimming lane.
Fig. 4 is the electrophorogram (part) that the Pi25 molecule marker primer utilizing the present invention to develop detects Cultivated Rice in China Mini core collection.
M:DL2000DNAmarker; P1, P2 are parent's Gumei2 that target SNP site is respectively C and G base
(Pi25) with 9311 (susceptible allelotrope), F1 is the first-filial generation of Gumei2 and 9311, and swimming lane 1-20 is respectively the portion of material of Cultivated Rice in China Mini core collection, and belonging to each material, genotype marks below corresponding swimming lane.
Fig. 5 is the F2 colony electrophorogram that the Pi25 molecule marker primer utilizing the present invention to develop detects the separation of target SNP site.
M:DL2000DNAmarker; P1, P2 are parent's Gumei2 that target SNP site is respectively C and G base
(Pi25) with 9311 (susceptible allelotrope), swimming lane 1-20 is the part F of the random selecting utilizing two parents to build
2individual plant, belonging to each material, genotype marks below corresponding swimming lane.
Embodiment
Experimental technique not specified in the present embodiment is molecular biology ordinary method.This institute Taq enzyme and dNTP originate from Beijing Quanshijin Biotechnology Co., Ltd, and all the other are routine biochemistry reagent.
Embodiment 1:
Rice blast resistant gene Pi 25 for rice special SNP codominant marker primer and detection method PCR condition optimizing
1) biomaterial
CC shaped material is the parent material Gumei2 containing Pi25 gene; GG shaped material is for containing susceptible allelic parent material 9311; CG shaped material is hybridize by Gumei2 and 9311 the F1 generation material obtained.
2) paddy DNA extracts and primer synthesis
CTAB method extracts the DNA of above-mentioned materials, and the primer sequence of synthesis shown in synthetic table 1, is specially:
Pi25-PFGGGATCTAGGCAGACAGT
Pi25-PRCAGAGCAGGAACTTCAGTTG
Pi25-NFAGCTGTGGCAAGCCTAACAG
Pi25-NRGATGCAGGATAAAGAATGAA
3)PCR
PCR reaction system is 15 μ L, containing 1.5 μ L10 × Buffer, and 1.0 μ LdNTPs (10mmol/L), 4 primer PF, PR, NF and NR are provided with 3 different concentration ratios and (are respectively 0.2 μM, 0.2 μM, 0.2 μM, 0.2 μM; 0.2 μM, 0.2 μM, 0.3 μM, 0.3 μM; 0.2 μM, 0.2 μM, 0.4 μM, 0.4 μM); Taq enzyme 0.2 μ L (5U/ μ L), 2.0 μ L template DNAs (50ng/ μ L), all the other use ddH
2o polishing; PCR response procedures: 94 DEG C of denaturation 5min; 94 DEG C of sex change 30s, arrange 3 different annealing temperature (54 DEG C, 56 DEG C, 58 DEG C) 40s, and 72 DEG C extend 40s, totally 35 circulations; 72 DEG C extend 5min, and amplified production is electrophoresis in 1.5% sepharose, by gel imaging instrument scanning record result.
4) interpretation of result
1, to be SNP site be 4,7 swimming lanes isozygotys the Pi25 shaped material of C base, utilize the present invention to mark specific band that amplification all has a 383bp in three kinds of annealing temperatures and three kinds of different primers concentration ratios; 3, to be SNP site be 6,9 swimming lanes isozygotys the susceptible allelotrope parent material of G base, all has the specific band of a 266bp in 56 DEG C and 58 DEG C of two kinds of annealing temperatures and two kinds of different primers concentration ratios; 2,5,8 for SNP site be the material of AG heterozygous base, in three kinds of annealing temperatures and three kinds of different primers concentration ratios, have 383bp and 266bp band respectively.
This result shows, the mark of our design is wide for primer thermal adaptability, is 56 DEG C in annealing temperature, band relatively clear (Fig. 2) when PF, PR, NF and NR concentration ratio are 1:1:1.5:1.5 amplification.
Embodiment 2:
The application of rice blast resistant gene Pi 25 for rice specific molecular marker primer, comprises the following steps:
To backbone parent amplification conventional in production:
1) biomaterial
Parent control is Gumei2 (Pi25), 9311 (susceptible allelotrope), Gumei2 and 9311 first-filial generation F1, test material is for producing upper conventional backbone parent: Mianhui725, bright extensive 63, Chinese Fragrant Rice, Japan are fine, osmanthus towards No. 2, Huang Huazhan, Lemont, B5, HP121, more light, the super Bath agate base of a fruit, salt rice No. 4, R1128, Peiai 64S, Guangzhan 63S, HD9802S, II-32A, golden 23A, the rich A in sky, huge wind 2A etc. 20, CTAB method extracts the DNA of above-mentioned materials.
2)PCR
PCR reaction system is 15 μ L, containing 1.5 μ L10 × Buffer, and 1.0 μ LdNTPs (10mmol/L), primer Pi25-PF and Pi25-PR is 0.2 μM, and primer Pi25-NF and Pi25-NR is 0.3 μ L; Taq enzyme 0.2 μ L (5U/ μ L), 2.0 μ L template DNAs (50ng/ μ L), all the other use ddH2O polishing; PCR response procedures: 94 DEG C of denaturation 5min; 94 DEG C of sex change 30s, 56 DEG C of annealing 40s, 72 DEG C extend 40s, totally 35 circulations; 72 DEG C extend 5min, and amplified production is electrophoresis in 1.5% sepharose, by gel imaging instrument scanning record result.
3) interpretation of result
Utilize special codominant marker's qualification in Pi25 gene of the present invention bright containing not homoallelic bill of material, Pi25 homozygote types of material has a 383bp specific band, and all the other allelic gene types are 266bp specific band, therefore the susceptible allelotrope of Pi25 resistant gene and all the other Commonly used parents can distinguish by this mark.As shown in Figure 3, the corresponding parent material of swimming lane 1-20 respectively is: extensive 725, bright extensive 63, the Chinese Fragrant Rice in another name for Sichuan Province, Japan are fine, osmanthus towards No. 2, Huang Huazhan, Lemont, B5, HP121, more light, the super Bath agate base of a fruit, salt rice No. 4, R1128, Peiai 64S, Guangzhan 63S, HD9802S, II-32A, golden 23A, the rich A in sky, huge wind 2A.P1, P2 be respectively parent's Gumei2 (Pi25 homozygote, SNP site is CC) and with 9311 (susceptible allele genic homozygote, SNP site is GG).Fig. 3 result shows, the genotype of its SNP site of paddy rice that swimming lane 1-20 identifies is G type, judges that it is susceptible allelotype, all conforms to actual.
Application process in the present embodiment, can be used for effectively carrying out genotype selection to blast resistant gene Pi 25 for rice, both may be used for the Screening and Identification of rice pest insects, may be used for again the molecular breeding of rice blast resistant gene Pi 25 for rice.
Embodiment 3:
The application of rice blast resistant gene Pi 25 for rice specific molecular marker primer, comprises the following steps:
Cultivated Rice in China Core Germplasms is increased:
4) biomaterial
Parent control is Gumei2 (Pi25), 9311 (susceptible allelotrope), Gumei2 and 9311 first-filial generation F1, test material is Cultivated Rice in China Mini core collection 200 parts.Cultivated Rice in China Mini core collection is that the kind of the representational kind of most 0.3% screened in more than 60,000 parts of Chinese rice germplasms saves the mark polymorphism of whole kind more than 70% and phenotypic polymorphism.CTAB method extracts the DNA of above-mentioned materials.
5)PCR
PCR system is with embodiment 2.
6) interpretation of result
Utilize special codominant marker in Pi25 gene of the present invention to identify that Cultivated Rice in China Mini core collection shows, in 200 kinds, occur that the bar the same with disease-resistant contrast Gumei2 is with 11, account for 5.5% of sum; The band that other 189 parts are all the same with susceptible variety 9311, accounts for 94.5% (partial results is shown in Fig. 4) of sum.Illustrate Pi25 gene at home in rice germplasm distribution frequency lower, the rice blast resistance improvement for China's rice varieties is provided convenient by newfound 11 kinds containing Pi25 resistance allele.
Application process in the present embodiment, can be used for effectively carrying out genotype selection to blast resistant gene Pi 25 for rice, both may be used for the Screening and Identification of Rice Germplasm Resources, may be used for again the molecular breeding of rice blast resistant gene Pi 25 for rice.
Embodiment 4:
The application of rice blast resistant gene Pi 25 for rice specific molecular marker primer, comprises the following steps:
The present embodiment is to rice blast resistant gene Pi 25 for rice F
2the detection of the single-gene separation of colony
1) biomaterial
P1, P2 are that target SNP site is respectively parent's Gumei2 (Pi25 homozygote) of C and G base and 9311 (susceptible allele genic homozygote), and Gumei2 (Pi25 homozygote) and 9311 (susceptible allele genic homozygote) hybridize the F built
2colony.
3)PCR
PCR system is with embodiment 2.
7) interpretation of result
P1, P2 be respectively parent's Gumei2 (Pi25 homozygote, SNP site is CC) and with 9311 (susceptible allele genic homozygote, SNP site is GG), F1 is the first generation of hybrid (SNP site is CG) of Gumei2 and 9311.
Utilize parent's Gumei2 (Pi25) and 9311 (susceptible allelotrope) to hybridize and construct F2 colony, by showing 93 individual plant genotype detection, the segregation ratio of 3 kinds of different genotype is 20CC:50CG:23GG, meets Mendelian's single-gene segregation ratio (χ of 1:2:1 through chi square test
2=0.72< χ
2 0.05=5.99), therefore this is labeled as codominant marker, and two kinds of different homozygotes and heterozygote can be distinguished, detection site shows as single-gene simultaneously and is separated (see Fig. 5).Swimming lane 1-20 is the part F of the random selecting utilizing two parents to build
2individual plant.
Application process in the present embodiment, can be used for effectively carrying out genotype selection to blast resistant gene Pi 25 for rice, both may be used for the Screening and Identification of rice pest insects, may be used for again the molecular breeding of rice blast resistant gene Pi 25 for rice.
SEQUENCELISTING
<110> Grain Crop Institute of Hubei Academy of Agricultural Sciences
Special SNP codominant marker exploitation and application in <120> rice blast resistant gene Pi 25 for rice gene
Special SNP codominant marker exploitation and application in <130> rice blast resistant gene Pi 25 for rice gene
<160>4
<170>PatentInversion3.1
<210>1
<211>18
<212>DNA
<213> artificial sequence
<400>1
gggatctaggcagacagt18
<210>2
<211>20
<212>DNA
<213> artificial sequence
<400>2
cagagcaggaacttcagttg20
<210>3
<211>20
<212>DNA
<213> artificial sequence
<400>3
gatgcaggataaagaatgaa20
<210>4
<211>20
<212>DNA
<213> artificial sequence
<400>4
agctgtggcaagcctaacag20
Claims (4)
1. special SNP codominant marker primer in rice blast resistant gene Pi 25 for rice gene, comprises Pi25-PF:GGGATCTAGGCAGACAGT; Pi25-PR:CAGAGCAGGAACTTCAGTTG; Pi25-NR:GATGCAGGATAAAGAATGAA; Pi25-NF:AGCTGTGGCAAGCCTAACAG.
2., to an authentication method for rice blast resistant gene Pi 25 for rice, comprising:
Utilize Pi25-PF:GGGATCTAGGCAGACAGT; Pi25-PR:CAGAGCAGGAACTTCAGTTG; Pi25-NR:GATGCAGGATAAAGAATGAA; Pi25-NF:AGCTGTGGCAAGCCTAACAG, carries out pcr amplification to paddy DNA; Pi25 genotype of isozygotying amplifies 612bp and 383bp band, and all the other allelic gene type amplified bands are 612bp and 266bp band, and the amplified band of heterozygous is 612bp, 283bp and 266bp band.
3. the application of primer according to claim 1 in rice blast resistant gene molecular breeding.
4. the application of primer according to claim 1 in the Screening and Identification of rice pest insects.
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| CN201610014679.9A CN105543366B (en) | 2016-01-11 | 2016-01-11 | Development and application of specific SNP codominant molecular marker in rice blast-resistant gene Pi25 gene |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109402236A (en) * | 2018-11-12 | 2019-03-01 | 长江大学 | The detection method of blast resistant gene Pi-25 in a kind of rice breed |
| CN112410459A (en) * | 2020-12-11 | 2021-02-26 | 上海市农业生物基因中心 | KASP molecular marker for detecting rice blast resistance gene Pi25 and application thereof |
| CN114736981A (en) * | 2022-03-18 | 2022-07-12 | 中国水稻研究所 | Functional specificity PCR molecular marker of rice blast resistance gene Pi25 and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101643790A (en) * | 2009-09-07 | 2010-02-10 | 中国水稻研究所 | Specific molecular marker of rice blast resistant gene Pi 25 for rice and special primer thereof |
| CN101824418A (en) * | 2009-12-10 | 2010-09-08 | 中国水稻研究所 | Coding region of rice blast resistant gene Pi 25 and application thereof |
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2016
- 2016-01-11 CN CN201610014679.9A patent/CN105543366B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101643790A (en) * | 2009-09-07 | 2010-02-10 | 中国水稻研究所 | Specific molecular marker of rice blast resistant gene Pi 25 for rice and special primer thereof |
| CN101824418A (en) * | 2009-12-10 | 2010-09-08 | 中国水稻研究所 | Coding region of rice blast resistant gene Pi 25 and application thereof |
Non-Patent Citations (1)
| Title |
|---|
| 王柯等: "PCR-CTPP：一种基于错配技术的SNP分型方法的改良", 《遗传》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109402236A (en) * | 2018-11-12 | 2019-03-01 | 长江大学 | The detection method of blast resistant gene Pi-25 in a kind of rice breed |
| CN112410459A (en) * | 2020-12-11 | 2021-02-26 | 上海市农业生物基因中心 | KASP molecular marker for detecting rice blast resistance gene Pi25 and application thereof |
| CN114736981A (en) * | 2022-03-18 | 2022-07-12 | 中国水稻研究所 | Functional specificity PCR molecular marker of rice blast resistance gene Pi25 and application thereof |
| CN114736981B (en) * | 2022-03-18 | 2023-10-31 | 中国水稻研究所 | Functional specificity PCR molecular marker of rice blast resistance gene Pi25 and application thereof |
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| CN105543366B (en) | 2020-02-07 |
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