CN105950747A - Rice blast resistance gene Pi1 functional specificity molecular marker and application thereof - Google Patents
Rice blast resistance gene Pi1 functional specificity molecular marker and application thereof Download PDFInfo
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Abstract
The invention provides a rice blast resistance gene Pi1 functional specificity molecular marker and application thereof. The molecular marker in a specificity banding pattern with a rice blast resistance gene Pi1 is amplified from a rice genome DNA through primer pairs of SEQ ID NO.1 and SEQ ID NO.2. The rice blast resistance gene Pi1 functional specificity molecular marker has very important application value, and by means of the marker, the utilization efficiency of the gene in germplasm resource screening, marker-assisted selection breeding, gene pyramiding breeding and transgenosis breeding can be improved.
Description
Technical field
The invention belongs to agricultural biological technical field, particularly to a kind of rice blast resistance genePi1Functional molecular marker
Pi1-InDel and method thereof and application.
Background technology
By Pyricularia oryzae (Magnapothe oryzae) the rice blast disease that causes is that Oryza sativa L. stable yields is endangered most important disease,
All can cause serious grain loss (Qu 1985, Ma et al. 2015) almost to the main producing region of world Oryza sativa L..Long-term life
Product practice have shown that, selection-breeding with utilize disease-resistant variety be preventing and treating rice blast the most safely and effectively method.Moreover due to rice blast diease occurrence
Reason microspecies pathogenicity variation is frequent, causes the resistance of single resistant variety gradually can lose in 35 years after planting
(the abundant institute etc. that obtains, 2011);Therefore, excavate and Appropriate application broad-spectrum resistance gene be obtain persistently, broad-spectrum disease resistance kind important
Approach.Conventional Resistance genes method has inoculated identification, but owing to different resistant genes have certain intercrossing in anti-spectrum,
Therefore traditional vaccination authentication method is not enough to accurately, reflects genotype veritably.In recent decades, along with paddy disease-resistant molecule
Genetic development, a lot of resistant genes are able to finely be positioned or clone (Su et al. 2015), and molecular marker
Development and application are greatly facilitated qualification and the development of many disease-resistant genes pyramiding breeding of resistant gene genetic background
(Hittalmani et al. 2000；Jena and Mackill, 2008).In the resistant gene cloned, some are disease-resistant
Gene is positioned at same site, sequence very high homology between these multiple alleless, between functional type sequence nand function type, and one
As linked marker be still difficult to accurately screen function disease-resistant gene (the Qu et al. 2006 of various material; Zhou et
al. 2006; Ashikawa et al. 2008; Takahashi et al. 2010; Yuan et al. 2011; Zhai
et al. 2011; Yuan et al. 2011; Hua et al. 2012;Ma et al.2015; Tian et al.
2016).Therefore, the sequence of direct analysis function allele itself develop the molecular marker of its specific Function and come target
Gene selects, and not only selects reliability high, also can be greatly accelerated breeding paces.
In recent years, it is related toPikThe research of the resistant gene of loci has had remarkable progress, at present on this resistance locus
The most at least it is found to have resistant gene (the Ashikawa et al. 2008 of 5 anti-spectrums of difference; Yuan et al. 2011;
Zhai et al. 2011;Hua et al. 2012)；Wherein, come from Oryza glaberrima Steud LAC23'sPi1Show wide spectrum high
Anti-characteristic (Mackill and Bonman 1992;Inukai et al. 1994), Oryza sativa L. main producing regions multiple to China
Rice blast microspecies all have the highest resistance (Hittalmani et al. 2000; Fuentes et al. 2008; Yang
et al.2008; Tacconi et al. 2010；Hua et al. 2012).Researcher exploitation go out some withPi1
The molecular marker of chain PCR-based technology carry out molecular marker assisted selection (MAS) breeding work (scholar's Liu equality, 2003；
Chen Zhiwei etc., 2005；Jin Sujuan etc., 2007；Jiang et al. 2012).But, these labellings used are all in resisting
Property genePi1Flank, there is certain physical distance with target gene, they are only used for the polymorphism between specific parent
Analyze, be not suitable for germplasm identification or Resistance resource screening.Pan Qinghua etc. (2012) are although developingPi1Specific Function
Molecular marker, but due to this SNP marker use before need identify K/N type genotype, in fact this genotype exist certain not
Correspondence, and qualification process needs the loaded down with trivial details work (Zhai et al. 2010) such as enzyme action qualification, is not suitable for extensive anti-
Property identify and MAS work.Therefore, in order to enable more accurately and effectively by rice blast broad-spectrum resistance genePi1It is applied to Oryza sativa L. resistance
In breeding work, it is necessary to exploitation truly reflect target gene, facilitate easy-to-useΡi1Special molecular marker.
Summary of the invention
In order to overcome the deficiencies in the prior art and shortcoming, the primary and foremost purpose of the present invention is to provide a kind of rice blast resistance genePi1Gene function specific molecular marker Pi1-InDel.
Another goal of the invention of the present invention is to provide described rice blast resistance genePi1Gene specific molecular marker
The detection method of Pi1-InDel.
Another goal of the invention of the present invention is to provide described rice blast resistance genePi1Gene function specific molecular
The application of labelling Pi1-InDel.
The purpose of the present invention is achieved through the following technical solutions: a kind of rice blast resistance genePi1Gene function specificity
Molecular marker Pi1-InDel, is to be amplified SEQ ID NO.1 and SEQ ID NO.2 from oryza sativa genomic dna by primer
With rice blast resistance genePi1Molecular marker in specificity banding pattern；
SEQ ID NO.1(5 '-3 '): GGTTGGTCGAAACCAGAAAA；
SEQ ID NO.2(5 '-3 '): GCTGAGGTAGAAGCGGGAGC；
Described rice varieties is IRBL1-CL(Pi1Donor parents)；
Described rice blast resistance genePi1Genetic fragment I including coding NBS-LRR albuminoid
I:GCTGAGGTAGAAGCGGGAGCCGAGGAGGCGACGCATCGGCGCCAAGGCGGAGGCGGAGGCAGAGTCGAGAG
CCGGGGAGACGGCACGCTGGCGCTGACGCAGAGGCGGAGGCGGCAGCCAGCGATGCGTGCTGCTGTGGCTAGTTTGT
GGCTAGTTCATGGCTACACCTCTCCCTAACCCTAGCTTAGCTCAGTGGCCTTTTTTCTGGTTTCGACCAACC 。
Described rice blast resistance genePi1The detection method of specific Function molecular marker Pi1-InDel, by than
To multiple resistance gene of rice blastPi1Allelic sequences, comprise the steps of
(1) download from public database and obtainPi1And the genome sequence of homology and order-checking kind Japan fine
(Nipponbare) genome sequence in corresponding region, forPi1Site carries out sequence alignment, examinationPi1Special, can district
Not in insertion/deletion (insertion-deletion, the InDel) site of other rice blast resistance alleles of this site；
(2) the InDel information that step (1) obtains is utilized, according to the design principle of InDel labelling, in described InDel site
Designing gene-specific primer at upstream and downstream 100-200 bp, primer is as follows to base sequence:
Fl:5- GGTTGGTCGAAACCAGAAAA -3；
Rl:5- GCTGAGGTAGAAGCGGGAGC -3；
(3) to carry rice blast resistance genePi1The STb gene of rice blast resistance kind IRBL1-CL be template, carry out PCR
Amplification, the PCR primer obtained is rice blast resistance genePi1Gene specific molecular marker Pi1-InDel；
Described rice blast resistance genePi1Gene specific molecular marker Pi1-InDel is differentiating that Rice Blast resists
Property gene application, be particularly suitable for differentiatePikThe application of the rice blast resistance gene that gene cluster region is different；Preferably comprise
Following steps:
(1) amplification: utilize primers F l and Rl that the genome of rice varieties to be detected is carried out PCR；
(2) detection: utilize polyacrylamide gel electrophoresis to detect, if the nucleotide sheet that molecular size range is 220 bp being detected
Section, then carry rice blast resistance genePi1；If the nucleotide fragments that molecular size range is 209 bp being detected, the most to be detected
Rice varieties carry the non-rice blast resistance gene in this sitePi1Other functional gene；If detecting, molecular size range is
265 bp or the nucleotide fragments being not detected by, rice varieties the most to be detected does not carriesPikThe functional gene in site.
The principle of the present invention: InDel refers to the same site of genome between sibling species or same species Different Individual
Sequence there occurs insertion or the disappearance of different size nucleotide fragments, and in i.e. one sequence, another of homology is compared in a certain site
Sequence is inserted or has lacked one or more base (Weber et al, 2002).InDel labelling PCR-based amplification technique, this
Length polymorphism labelling (Hyten et al, 2010) is belonged in matter.InDel labelling good stability, polymorphism are high, classification system
Simply (Jander et al, 2002)；Compared with the SNP marker complicated with classification system, InDel detection is simpler convenient, right
Instrument and equipment and technology require relatively low, can carry out on electrophoretic techniques platform.Have started to be applied to animals and plants colony at present lose
Pass the fields such as analysis, marker assisted selection and mankind's medicolegal genetics, medical diagnosis.The present invention side by Multiple Sequence Alignment
Method is foundPi1The InDel occurred in gene order, owing to this InDel existsPi1WithPikOther functional gene have 11 bp's
Difference, Japan fine in do not contain this site, be therefore easy to byPi1Therefrom distinguish.The present invention designs primer pair accordingly
F1/R1, by standard PCR amplification, when polyacrylamide gel electrophoresis detects, segment in different sizes is presented.
The present invention has such advantages as relative to prior art and effect:
(1) the molecular marker specificity that the present invention provides is high:Pi1The genome area at place also exists and includesPi1Interior 5
The individual candidate gene with resistant gene feature, these candidate genes are very high homology (Zhai et al. 2010) in sequence；
Additionally,Pi1-5CWith the rice blast resistance gene cloned on this sitePikm-1、Pik-1WithPikp-1At amino acid levels
On sequence identity be respectively 99%, 99% and 95%,Pi1-6CWithPikm-2,Pik-2WithPikp-2On amino acid levels
Sequence identity be all 99% (Hua et al. 2012).The molecular marker that the present invention provides is that inventor passes through
Constantly carry out sequence polymorphism seeking and obtained by experimental verification, can significantly byPi1Be present in this genome district
In territory withPi1The paralog gene of very high homology makes a distinction；Also can successfully distinguishPik-m、Pik 、Pik-p、Pik-s、Pi7WithPik-hWith other non-functional homologous genes.
(2) molecular marker that the present invention provides is in actual applications, low cost, high flux: be to utilize electrophoresis mostly at present
Platform carries out typing, and this typing platform is the most economical, it is not necessary to complicated experimental facilities, workable.Electropherotyping platform
There are agarose gel electrophoresis, degeneration or native polyacrylamide gel electrophoresis and capillary electrophoresis.What the present invention provided divides
Sub-labelling only needs PCR to combine agarose gel electrophoresis or native polyacrylamide gel electrophoresis, and low cost, flux are high, add
Specificity high (i.e. accuracy is high), is particularly well-suited in production practices.
(3) present invention be first forPi1Gene internal sequence is developedPi1Gene specific InDel labelling.This
Invention can successfully will by the method for electrophoresis detectionPi1Be positioned on this site other rice blast resistance gene (Pikm,Pik,Pikp,Pik-s,Pi7WithPikh) distinguish, up to the present, also there is no the report about this kind of labelling.Institute of the present invention
There is providedPi1Specific molecular marker Pi1-InDel is a codominant marker, its reliability and standard in actual application
Really property is better than dominant marker.Present invention can apply toPi1Screening of Germplasm, transgenic identify and gene pyramiding and based on
In the Oryza sativa L. resistance breeding work of MAS technology.This labelling is present inPi1Gene internal, the most rightPi1Screening capacity theoretical value
Up to 100%, its combination property be better than having reported withPi1Chain molecular marker and functional label.
(4) molecular marker that the present invention provides can be applied in the colony that genetic background is different easily.Existing withPi1
Chain molecular marker major part is both for what the sequence polymorphism of same 2 different parents of colony was developed, these marks
The note suitability in other colony is limited.The functional label of the exploitation such as Pan et al (2012), owing to qualification process compares
Loaded down with trivial details, it is not suitable in large-scale germplasm identification and MAS breeding.What the present invention was applicable under any genetic background turns base
Because of breeding, gene pyramiding and resistance breeding based on MAS technology, it is not necessary to repeat the screening of parent's polymorphism, significantly carry
High breeding efficiency.
Therefore, rice blast resistance gene provided by the present inventionPi1Gene specific molecular marker has important application
It is worth, utilizes this labelling can improve this gene and educate at Screening of Germplasm, molecular marker assisted selection breeding, gene pyramiding
Kind, and the efficiency utilized in transgenic breeding.
Accompanying drawing explanation
Fig. 1 isPi1Gene specific molecular marker Pi1-InDel existsPikGene cluster region difference rice blast resistance gene
And fine, the result figure of 9311 of Japan, wherein: swimming lane M is DNA ladder, the DNA profiling of swimming lane 19 is followed successively by resistance
Kind IRBL1-CL (Pi1), resistant variety IRBLk-Ka (Pik), resistant variety IRBLkp-K60 (Pik-p), resistant variety
IRBLkm-Ts (Pik-m), resistant variety IRBLkh-K3 (Pik-h), resistant variety IRBLks-S (Pik-s), resistant variety
IRBL7-M (Pi7), susceptible variety Japan fine (NPB), susceptible variety 9311.
Fig. 2 isPi1Gene specific molecular marker Pi1-InDel the result figure in other rice varieties, wherein:
Swimming lane M1 is DNA ladder, the DNA profiling of swimming lane 1-12 be followed successively by resistant gene donor kind IRBL1-CL (Pi1), precious Shan
97B, Co39, Fuhui 838, Gu Feng B, Jin Nante 43B, special number of south, pacify rich B, special blue or green No. 2, bag association 123B, Nanjing 11, bamboo precious
B, short-foot Nan Te.
Detailed description of the invention
Below in conjunction with embodiment and accompanying drawing, the present invention is described in further detail, but embodiments of the present invention do not limit
In this.
Embodiment 1: rice blast resistance genePi1Gene specific molecular marker and design of primers thereof and detection
(I) analysis in Pi1 insertion/deletion (insertion-deletion, InDel) site:
Download from public database and obtainPi1AndPik-m,Pik,Pik-p,Pik-s,Pi7WithPik-hOryza sativa L. donor kind
Partial genome sequence, order-checking kind Japan fine (Nipponbare) and 9311 without the genome sequence in corresponding region, for
Pi1 site carries out sequence alignment, examinationPi1Special, other rice blast resistance alleles of this site can be different from special
Property insertion/deletion (InDel) difference site.There is the resistance gene of rice blast kind of gene specific InDel
IRBL1-CL (Pi1) and resistant variety IRBLk-Ka (Pik), resistant variety IRBLkp-K60 (Pik-p), resistant variety
IRBLkm-Ts (Pik-m), resistant variety IRBLkh-K3 (Pik-h), resistant variety IRBLks-S (Pik-s) and resistance product
Kind IRBL7-M (Pi7) Multiple Sequence Alignment result following table shown in:
Wherein, add the nucleotide of red overstriking to be and identifyPi1Gene specific insertion sequence；
(2) design primer:
According to the design principle of InDel labelling, at described InDel site upstream and downstream 100-200 bp, design primer, primer
As follows to base sequence:
Fl: 5′-GGTTGGTCGAAACCAGAAAA-3′；
Rl: 5′-GCTGAGGTAGAAGCGGGAGC-3′。
(3) Oryza sativa L. Representative Cultivars is selected: select to carryPi1Gene andPikThe allelic Representative Cultivars of gene cluster
As follows:
Rice varieties IRBL1-CL, forPi1Donor kind (Fukuta et al. 2004), for positive control；
Rice varieties IRBLk-Ka, forPikDonor kind (Fukuta et al. 2004)；
Rice varieties IRBLkp-K60, forPik-pDonor kind (Fukuta et al. 2004)；
Rice varieties IRBLkm-Ts, forPik-mDonor kind (Fukuta et al. 2004)；
Rice varieties IRBLkh-K3, forPik-hDonor kind (Fukuta et al. 2004)；
Rice varieties IRBLks-S, forPik-sDonor kind (Fukuta et al. 2004)；
Rice varieties IRBL7-M, forPi7Donor kind (Fukuta et al. 2004)；
Susceptible check variety: Japan fine, Japan fine be Japan selection-breeding a kind, country Oryza sativa L. data center
(http://www.ricedata.cn/variety/varis/602979.htm ?602979) relevant information can be obtained.
Susceptible check variety: 9311, selection-breeding unit: Inst. of Agricultural Science, Lixiahe Prefecture, Jiangsu Prov. (http: //
www.ricedata.cn/variety/varis/600611.htm)。
(4) PCR amplification, it is thus achieved that containPi1The fragment of gene specific InDel
Utilize above-mentioned primer to Fl and R1, with the STb gene of above-mentioned rice varieties as template, gather after carrying out standard PCR amplification
Acrylamide gel electrophoresis detects, and the result obtained is as shown in Figure 1.
Amplification reaction system is as follows:
2 x Reaction Mix: 12.5 μL
Primers F l (10 μ Μ): 1 μ L
Primer Rl (10 μ Μ): 1 μ L
Golden DNA Polymerase: 0.2 μL
DNA template (20-50 ng/ μ L): 1 μ L
ddH2O: complement to 25 μ L.
PCR Thermal cycling conditions is as follows: 94 DEG C 5 minutes；94 DEG C 30 seconds, 58 DEG C 30 seconds, 72 DEG C 30 seconds, 35 are followed
Ring；72 DEG C 7 minutes；10 DEG C of preservations.
After PCR reaction terminates, take appropriate amount of sample in the polyacrylamide gel of 8 %, carry out electrophoresis detection, deposition condition
It is 90 V, 1 hour.
Wherein, the swimming lane I of Fig. 1 isPi1Donor kind rice varieties IRBL1-CL genome is the sheet that template PCR obtains
Section, with rice blast resistance genePi1In specificity banding pattern, i.e. swimming lane 1 isPi1Gene specific molecular marker Pi1-InDel.
The result of Fig. 1 shows,Pi1Gene specific molecular marker can distinguish anti-sense allele, can distinguish againPikReflected on site
Surely other resistant gene arrived.Carry it is to say, everyPi1Rice varieties, the electrophoresis detection bar of its pcr amplification product
Band presents with 220 bp,PikOther rice blast resistance gene positioned on site presents with electrophoresis detection band 209 bp,
And the functional gene not containing this site with electrophoresis detection band 265 bp or presents without band.
Embodiment 2: resistant genePi1Gene specific molecular marker is differentiatingPikGene cluster region difference rice blast resists
The application of property gene.
PCR primer band according to polymorphism, can be containing target genePi1With otherPikResistance in gene cluster
Gene regions separates.As it is shown in figure 1, can be by according to pillar locationPi1Other resistant gene district identified with this site
Separate, 220 bp represent containingPi1Gene, 209 bp then represent other resistant gene containing this site, other banding pattern table
Show the functional gene not containing this site.Visible result of the test and matching that design is analyzed, illustrate resistant genePi1Gene is special
Opposite molecule labelling can differentiatePi1Gene withPikThe aspects such as gene cluster allele are applied.
Embodiment 3: resistant genePi1The detection application in other rice varieties of the gene specific molecular marker
Select 12 representative rice varieties, be followed successively by: Zhenshan 97B, Co39, Fuhui 838, Gu Feng B, Jin Nante 43B, Nan Te
Number, pacify rich B, special blue or green No. 2, bag association 123B, Nanjing 11, bamboo treasure B, short-foot Nan Te.
Extract its genomic DNA respectively, and as template, according to the method for embodiment 1, carry out PCR amplification, enzyme action and
Electrophoresis detection.According to the size of digestion products (band), can be resistant genePi1Distinguish with other rice blast resistance genes
Come.As in figure 2 it is shown, in anti-stave type bePi1The individuality of type (carries rice blast resistance genePi1Disease-resistant variety IRBL1-
CL, the sample of swimming lane 1) in the existence of Pi1-InDel gene specific molecular marker can be detected, and other anti-stave types
Individuality then can not detect this molecular marker.Visible, result of the test and matching that design is analyzed, resistant gene is describedPi1Base
Because specific molecular marker can differentiatePi1Gene and other rice blast resistance genes, includingPikThe side such as gene cluster allele
Face is applied.
Above-described embodiment is the present invention preferably embodiment, but embodiments of the present invention are not by above-described embodiment
Limit, the change made under other any spirit without departing from the present invention and principle, modify, substitute, combine, simplify,
All should be the substitute mode of equivalence, within being included in protection scope of the present invention.
SEQUENCE LISTING
<110>Fujian Province Agriculture Science Academy, Institute of Biotechnology
<120>a kind of rice blast resistance gene Pi 1 specific Function molecular marker and application thereof
<130> 10
<160> 10
<170> PatentIn version 3.3
<210> 1
<211> 20
<212> DNA
<213>artificial sequence
<400> 1
ggttggtcga aaccagaaaa 20
<210> 2
<211> 20
<212> DNA
<213>artificial sequence
<400> 2
gctgaggtag aagcgggagc 20
<210> 3
<211> 220
<212> DNA
<213>genetic fragment I
<400> 3
gctgaggtag aagcgggagc cgaggaggcg acgcatcggc gccaaggcgg aggcggaggc 60
agagtcgaga gccggggaga cggcacgctg gcgctgacgc agaggcggag gcggcagcca 120
gcgatgcgtg ctgctgtggc tagtttgtgg ctagttcatg gctacacctc tccctaaccc 180
tagcttagct cagtggcctt ttttctggtt tcgaccaacc 220
<210> 4
<211> 54
<212> DNA
<213> IRBL1-CL -Pi1
<400> 4
gccagcgatg cgtgctgctg tggctagttt gtggctagtt catggctaca cctc 54
<210> 5
<211> 43
<212> DNA
<213> IRBLk-Ka-Pik
<400> 5
gccagcgatg cgtgctgctg tggctagttc atggctacac ctc 43
<210> 6
<211> 43
<212> DNA
<213> IRBLkp-K60-Pik-p
<400> 6
gccagcgatg cgtgctgctg tggctagttc atggctacac ctc 43
<210> 7
<211> 43
<212> DNA
<213> IRBLkm-Ts-Pik-m
<400> 7
gccagcgatg cgtgctgctg tggctagttc atggctacac ctc 43
<210> 8
<211> 43
<212> DNA
<213> IRBLkh-K3-Pik-h
<400> 8
gccagcgatg cgtgctgctg tggctagttc atggctacac ctc 43
<210> 9
<211> 43
<212> DNA
<213> IRBLks-S-Pik-s
<400> 9
gccagcgatg cgtgctgctg tggctagttc atggctacac ctc 43
<210> 10
<211> 43
<212> DNA
<213> IRBL7-M-Pi7
<400> 10
gccagcgatg cgtgctgctg tggctagttc atggctacac ctc 43
Claims (6)
1. a rice blast resistance genePi1The primer pair of gene function specific molecular marker, it is characterised in that: described primer
It is SEQ ID NO.1(5 '-3 ' to sequence): GGTTGGTCGAAACCAGAAAA；
SEQ ID NO.2(5 '-3 '): GCTGAGGTAGAAGCGGGAGC.
2. a rice blast resistance genePi1Gene function specific molecular marker, it is characterised in that: it is to SEQ by primer
ID NO.1 and SEQ ID NO.2 amplifies and rice blast resistance gene from oryza sativa genomic dnaPi1In specificity banding pattern
Molecular marker.
A kind of rice blast resistance gene the most according to claim 2Pi1Gene function specific molecular marker, its feature exists
In: described rice blast resistance genePi1Including genetic fragment I of coding NBS-LRR albuminoid, I sequence such as SEQ ID NO.3
Shown in.
4. rice blast resistance gene as claimed in claim 2Pi1The detection method of specific Function molecular marker, its feature
It is: by the multiple resistance gene of rice blast of comparisonPi1Allelic sequences, comprise the steps of
(1) download from public database and obtainPi1And the genome sequence of homology and order-checking kind Japan fine relatively
Answer the genome sequence in region, forPi1Site carries out sequence alignment, examinationPi1Special, other rice of this site can be different from
Insertion/deletion (insertion-deletion, the InDel) site of pestilence resistance allele；
(2) the InDel information that step (1) obtains is utilized, according to the design principle of InDel labelling, in described InDel site
Designing gene-specific primer at upstream and downstream 100 200bp, primer is as follows to base sequence:
Fl:5- GGTTGGTCGAAACCAGAAAA -3；
Rl:5- GCTGAGGTAGAAGCGGGAGC -3；
(3) to carry rice blast resistance genePi1The STb gene of rice blast resistance kind IRBL1-CL be template, carry out PCR
Amplification, the PCR primer obtained is rice blast resistance genePi1Gene specific molecular marker Pi1-InDel.
5. rice blast resistance gene as claimed in claim 2Pi1Gene specific molecular marker is differentiating rice varieties rice blast
The application of sick resistant gene.
Rice blast resistance gene the most according to claim 2Pi1Gene specific molecular marker is differentiatingPikGene cluster
The application of the rice blast resistance gene that region is different.
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