CN103409433A - Novel broad-spectrum rice blast resistance allele pi21t and resistance application thereof - Google Patents
Novel broad-spectrum rice blast resistance allele pi21t and resistance application thereof Download PDFInfo
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Abstract
The invention belongs to the field of molecular genetics and relates to a novel broad-spectrum rice blast resistance allele pi21t and resistance application thereof. According to a cloned sequential design primer of the broad-spectrum rice blast resistance recessive allele pi21t, PCR amplification is performed on the wide compatibility variety 02428 which is used in high frequency in breeding, and sequencing comparative analysis is performed, so that the novel broad-spectrum rice blast resistance allele pi21t is identified in combination with a field rice blast inoculation identification result of the variety. The material has high resistance for popular microspecies of 6 rice blast groups in the Yangtze-Huaihe river basin, and provides new gene resources for breeding for rice blast resistance.
Description
Technical field
The present invention has identified the neomorph of a wide spectrum blast resisting
Pi21tAnd the resistance application, belong to the molecular genetics field, be exclusively used in the Molecular design breeding of rice anti-rice blast, to for the breeding of paddy rice wide spectrum blast resisting, providing new resource, by the molecular marker assisted selection breeding, accelerate the process of paddy rice wide spectrum blast resisting breeding, improve disease-resistant efficiency.
Background technology
Paddy rice is one of most important food crop in world wide, is also the first food crop of China, improves rice yield and ultimate production and has very important effect for the grain security that ensures China.Rice blast is most important worldwide disease on paddy rice, is commonly called as paddy rice " cancer ", more than 80 countries of extend over the entire globe, in China, generation is in various degree arranged every year often, the general underproduction 10～20% in grave illness district, the serious time underproduction can reach 40～50%, and even No kernels or seeds are gathered, as in a year of scarcity in partial area.Since the nineties in 20th century, Chinese rice blast generation area every year is all at 3,800,000 hm
2Above, year loss paddy reaches several hundred million kilograms, has become the significant obstacle that paddy rice continues high yield, stable yields.Facts have proved, control the most economical effective means of rice blast and be exactly seed selection and utilize disease-resistant variety.Therefore, the discovery of disease-resistant gene is vital with the clone to carrying out smoothly of rice blast breeding for disease resistance.
Rice blast be by Pyricularia oryzae (
Magnaporthe grisea Barr, asexual generation is
Pyricularia grisea Sacc) infect and cause, the rice blast fungus microspecies have the variability of height, sustainable utilization along with resistant gene, the pathogenic bacterium colony genetic construction can constantly change, new physiological strain appears, thereby cause race specific resistance to be lost, most resistant varieties are lost disease resistance gradually at several Nian Houhui of plantation.Therefore, need to constantly excavate and identify new resistant gene, and clone and utilize the broad spectrum durable resistant gene is breeding rice wide spectrum blast resisting new variety, solves the effective way of rice blast harm.
Up to now, identify in the world a large amount of anti-sources of rice blast wide spectrum, and the anti-source of part has been carried out location and the clone of anti-pest gene.Anti-pest kind Moroberekan as lasting as internationally recognized Africa, lose its high resistance for many years and not at the West Africa establishing in large scale, and the high resistance of its rice blast is mainly because this kind has comprised 3 Major resistance genes and 10 minor effect resistance sites.At present, successfully clone the anti-pest gene of a plurality of wide spectrums, as
Pib,
Pita,
Pi9,
Piz,
Pi21Etc..
PibBe by Japanese scholars clone's the 1st rice blast resistance gene, belong to the NBS-LRR disease-resistant gene member of family, be subjected to the abduction delivering of temperature and dark condition.
Pi9Be to have cloned at present the widest one of anti-spectrum in anti-seasonal febrile diseases gene, coded product has the NBS-LRR structural domain equally.
Pi21It is up-to-date clone's an anti-seasonal febrile diseases gene of non-microspecies specificity wide spectrum, different with the constitutional features of the anti-seasonal febrile diseases gene of cloning before, its coding be a proline rich albumen, comprise the heavy metal binding structural domain of a supposition and the interactions between protein motif of a supposition.Afunction after this transgenation disappearance, even reach immunity thereby make resistance improve, and through identifying, finds that some japonica rice varieties have carried the resistance allele of this gene, for the rice anti-rice blast breeding provides favourable resource.The clone of above these genes makes the anti-pest Molecular design breeding of paddy rice wide spectrum become possibility.
The present invention is according to the rice blast broad-spectrum disease resistance Recessive alleles of having cloned
Pi21Primers, the wide affine kind 02428 that high frequency in breeding is used is carried out to pcr amplification, the order-checking comparative analysis, in conjunction with field rice blast inoculated identification result, identified the neomorph of a wide spectrum blast resisting
Pi21t, can be used as the new resources of paddy rice wide spectrum blast resisting breeding, by the molecular marker assisted selection breeding, accelerate the process of paddy rice wide spectrum blast resisting breeding, improve disease-resistant efficiency.
Summary of the invention
Technical problem
The objective of the invention is: according to the rice blast broad-spectrum disease resistance Recessive alleles of having cloned
Pi21(GenBank:AB430853.1, http://www.ncbi.nlm.nih.gov/nuccore/AB430853.1) primers, the wide affine kind 02428 that high frequency in breeding is used is carried out to pcr amplification, the order-checking comparative analysis, in conjunction with field rice blast inoculated identification result, identified the neomorph of a wide spectrum blast resisting
Pi21t, be mainly the breeding of paddy rice wide spectrum blast resisting new favourable resource be provided.
Technical scheme
A rice blast broad-spectrum disease resistance neomorph
Pi21t, it is characterized in that: this equipotential gene
Pi21tTotal length is 874bp, and its sequence is:
ATGGGTATATTGGTCATCTTGGTGGACCTGCAATGCTGCCGCTGCGATGCCAAGATCAGGAAGGTCCTGGGCTGCCTTGAAGGTATAATAAATTCTGCCCGAATCGTCCATGTTTGATTGAATTTTCAAGGCTAATCAGCAGTGTTCCTGCTCAATTGGGAGCAAAACCTCTGTTAAAAAGGGTGTGTTTGAATGAATATAATTGAATATGAACGCAGAGGAGTACTGCATCGAGAAGGTGGAGTACGACGTGAAGAACAACAGGGTGATCGTGCGCGGGAAGTTCGACCCGGAGAAGCTGTGCAAGAAGATCTGGTGCAAGGCCGGCAAGATCATCAAGGAGATCCTCATCGTCGACGTCTGGCCGCCGCCGCCGCCGTGCGAGAAGCCTCCGGAGGACTGCAAGCCCAAGCCCTGCCATTGCTGCAGCTGCGAGAAGCCCAAGCCCAAGCCCAAGCCCTGCCACTGCGAGAAGCCCAAGCCCTGTCACTGCGAGAAGCCCAAGCCATGCGAGAAGCCGCCGCCGGAGAAGCCGCCGCCGAAGCCGGAGTGCAAGCTGGTGCCGTACCCTTACCCGGTGCCGTACCCGTACGCCGGGCAGTGGTGCTGCCCAAAGCCTGAGCCGCCGAAGCCGCCGCCGGAGCCACCGAAGGAGCCGGAGCCGCCGAAGCCGTGCGGGTGCTCGCACGCCTTCGTGTGCGTCTGCAAGCCGGCGCCGCCGCCGCCGCCGCCGTGCGGGTGCTCGGGGGGCCACGGGAACTGCGGCTGCGGCATCAGGCCGTGGCCGCCGCAGGTGTGGCCGCCGCCGCCCGTCTGCCCGCCGCCGCCGTGGTGCTACACCGAGGACAACGCCAACGCCTGCTCCATCATGTGA。
This equipotential gene
Pi21tFrom wide affine kind 02428,6 groups' of rice blast of Yangtze-Huaihe River Valley Major Epidemic Races is all shown as to high resistance, with respect to susceptible allelotrope
Pi21Have two place's sequence deletions, its deletion sequence is respectively: GCCGCTGCCGCAGCCGCCGCCGCCGTGCAA and GAAGCCGCCGCCGTGCAAGCCGGAGGAGCCGCC.
Described neomorph
Pi21tThe amino acid of deriving, its sequence is:
MGILVILVDLQCCRCDAKIRKVLGCLEEEYCIEKVEYDVKNNRVIVRGKFDPEKLCKKIWCKAGKIIKEILIVDVWPPPPPCEKPPEDCKPKPCHCCSCEKPKPKPKPCHCEKPKPCHCEKPKPCEKPPPEKPPPKPECKLVPYPYPVPYPYAGQWCCPKPEPPKPPPEPPKEPEPPKPCGCSHAFVCVCKPAPPPPPPCGCSGGHGNCGCGIRPWPPQVWPPPPVCPPPPWCYTEDNANACSIM-。
Described neomorph
Pi21tWith the amino acid of its derivation, can in the rice blast broad-spectrum disease resistance, be applied.
Beneficial effect
According to the rice blast broad-spectrum disease resistance Recessive alleles of having cloned
Pi21The primers of (GenBank:AB430853.1, http://www.ncbi.nlm.nih.gov/nuccore/AB430853.1), sequence is as follows:
Pi21-1F: 5’ GGTTATCAAGACCCCTCCTG 3’
Pi21-1R: 5’ AGAAGGGGAGAAACGAAGAG 3’
PCR product size is 1915bp in dominant allele, the order-checking compare of analysis, in Recessive alleles, the PCR product exists 48bp and 21bp two place's disappearances, consistent with the result of having announced, illustrate and utilize the different rice varieties of this primer pair sequencing analysis that increases to can be used to identify the different allelotypes of target gene.
Utilize the wide affine kind 02428 that on above-mentioned primer pair rice breeding, high frequency is used to carry out pcr amplification, the order-checking compare of analysis, found to be different from disappearance type (Fukuoka S, the Saka N of the 30bp+33bp of the existing deletion allele type of having delivered, Koga H
Et al. Loss of Function of a Proline-Containing Protein Confers Durable Disease Resistance in Rice. Science, 2009,325:998-1001), illustrate that 02428 has carried a rice blast broad-spectrum disease resistance neomorph, called after
Pi21t.This equipotential gene
Pi21tFrom wide affine kind 02428,6 groups' of rice blast of Yangtze-Huaihe River Valley Major Epidemic Races is all shown as to high resistance, with respect to susceptible allelotrope
Pi21Have two place's sequence deletions, its deletion sequence is respectively: GCCGCTGCCGCAGCCGCCGCCGCCGTGCAA and GAAGCCGCCGCCGTGCAAGCCGGAGGAGCCGCC.
Described allelotrope
Pi21tCan be used in rice blast broad-spectrum disease resistance breeding.The broad-spectrum disease resistance neomorph of the authentication method of rice blast allelotype provided by the present invention and evaluation
Pi21t, have following advantage:
By the primer pair target variety designed in the present invention, carry out pcr amplification, reclaim product order-checking compare of analysis, the allelotype of entrained target gene in the hard objectives kind fast.In the present invention, identified that simultaneously the wide affine kind 02428 that on the rice breeding, high frequency is used carries rice blast broad-spectrum disease resistance neomorph
Pi21tWide affine kind 02428 is carried out to field rice blast inoculated identification, and strains tested is Jiangsu Province's Pyricularia oryzae representative strain of separating to obtain in 2009 totally 6 microspecies, and seedling stage, qualification result was 0 grade, namely show as immunity, for the breeding of paddy rice wide spectrum blast resisting provides new favourable resource.
The accompanying drawing explanation
Target allelotrope in the wide affine kind 02428 of Fig. 1
Pi21tThe agarose gel electrophoresis result of pcr amplification product 1%.
Target allelotrope in the wide affine kind 02428 of Fig. 2
Pi21tPcr amplification product reclaim after 1% agarose gel electrophoresis detected result.
In the wide affine kind 02428 of Fig. 3 the allelic partial sequence of target and with dominant susceptible allelotrope
Pi21Comparison result.The dominant susceptible allelotrope that in figure, the Pi21 representative has been announced
Pi21Partial sequence, J2-15 represents the allelic partial sequence of target in 02428.
The agarose gel electrophoresis result of the Partial Fragment pcr amplification product 3% of the wide affine kind 02428 of Fig. 4 and check variety Suyunuo target gene.1 is 02428,2 to be Suyunuo.
Embodiment
In following embodiment, method therefor is ordinary method if no special instructions.
(1) the wide affine kind 02428 rice blast broad-spectrum disease resistance neomorph of carrying
Pi21tAuthentication method
1, the extraction of wide affine kind 02428 genomic dna
Get wide affine kind 02428 plant young leaflet tablet, extract oryza sativa genomic dna by 2 * CTAB method (100mmol/L Tris-Cl, 20mmol/L EDTA, pH 8.0 for 2%CTAB, 1.4mol/L NaCl).
The rice leaf of about 0.1g is positioned in the little centrifuge tube of 2mL, with bamboo let, smashes powdered after liquid nitrogen freezing; Add 700 μ L to extract buffered soln through 2 * CTAB of 65 ℃ of preheatings, fully mix and be placed on 65 ℃ of water-bath 30min, mix once every 5min; After water-bath, in each centrifuge tube, add 700 μ L chloroforms, turn upside down and fully mix, extracting 5min; 10000rpm, centrifugal 8min, get supernatant liquor 500 μ L and be transferred in the little centrifuge tube of 1.5mL; Add 500 μ L freezing Virahol in advance, standing 30min on ice after mixing; 12000rpm, centrifugal 5min; Abandon most supernatant, the DNA precipitation is washed 2 times with 70% alcohol; Abandon most washings, be dissolved in the sterilizing distilled water of 200 μ L after the DNA precipitation is air-dry, save backup in-20 ℃.
2, PCR reaction and electrophoretic analysis
As above, according to the rice blast broad-spectrum disease resistance Recessive alleles of having cloned
Pi21The primers of (GenBank:AB430853.1, http://www.ncbi.nlm.nih.gov/nuccore/AB430853.1), carry out pcr amplification to wide affine kind 02428 plant DNA, and primer sequence is as follows:
Pi21-1F：5’ GGTTATCAAGACCCCTCCTG 3’
Pi21-1R：5’ AGAAGGGGAGAAACGAAGAG 3’
The long segment amplification system is: DNA 2.0 μ L, 10 * LA Buffer (Mg
2+Plus) 5.0 μ L, dNTP 8.0 μ L, primer 2 .0 μ L, LA Taq enzyme 0.5 μ L, add ddH
2O supplies 50 μ L.
Pcr amplification program: 95 ℃ of denaturation 5min; 94 ℃ of sex change 30sec, 55 ℃ of annealing 30sec, 72 ℃ are extended 2min (1min/Kb), 33 circulations; 72 ℃ of insulation 5min; 18 ℃, insulation.
Electrophoretic analysis: to the PCR product of short duration centrifugal after, in each reaction tubes, add 3 μ l sample-loading buffers.Preparation 1% sepharose, take in the triangular flask that the 1g agarose joins 100 ml 1 * TAE damping fluids, fully boils in microwave oven, shakes up gently, after it is slightly cooling, adds 1～2 EB solution, records the glue plate after shaking up gently.After isogel was cooling, (approximately 30min) pulled out comb and puts into electrophoresis chamber, can the loading electrophoresis.Voltage with 8～12 V/cm in 1 * TAE damping fluid carries out electrophoresis, when the gel forward position is arrived in the swimming of indicator tetrabromophenol sulfonphthalein band, finish electrophoresis, utilize the Bio-Rad gel imaging system take pictures and investigate amplification, see Fig. 1, obtain being about the target fragment of 2.0kb size, size is correct, prepares to cut glue and reclaims.
3, in sepharose, reclaim target dna fragment (Takara Kit) and sequencing analysis (Invitrogen)
The target dna fragment reclaims: after agarose gel electrophoresis is analyzed, to the rubber tapping of DNA target stripe, insert in 1.5ml Eppendorf pipe, and weigh.The 1mg agarose gel is equivalent to 100 μ l volumes; The Buffer DE-A that adds 3 volumes of agarose gel to the Eppendorf pipe, bathe 10min 65 ℃ of temperature after mixing, and is interrupted and mixes, until blob of viscose melts fully; Buffer DE-B to adding 0.5 Buffer DE-A volume in every pipe, mix; 12,000rpm, of short duration (10s) is centrifugal, abandons supernatant; Add 500 μ l Buffer W1, of short duration centrifugal under 12,000rpm, abandon supernatant; Add 700 μ l Buffer W2, of short duration centrifugal under 12,000rpm, abandon supernatant; Repeat previous step once; Under 12,000rpm of short duration sky from; By preparing pipe, put into clean 1.5ml centrifuge tube, add 30 μ l preheating ddH in film central authorities
2O, standing 1min, the of short duration centrifugal eluted dna of 12,000rpm; Utilize agarose gel electrophoresis to test to reclaiming fragment, clip size is correct, sees Fig. 2.
Sequencing analysis: send company to check order (Invitrogen) the target dna fragment reclaimed, sequencing result and the dominant susceptible allelotrope of having announced
Pi21The sequence alignment analysis, find to have two places' disappearances, be respectively 30bp and 33bp (Fig. 3), this allelic disappearance type with delivered the deletion allele type all not identical, be a new allelotrope, the called after neomorph
Pi21t.
(2) wide affine kind 02428 Field inoculation authentication method and seedling stage qualification result
1,02428 pair of blast resistance identification method of wide affine kind
The strains tested of blast resistance identification is Jiangsu Province's Pyricularia oryzae representative strain that separation in 2009 obtains: 2010-49-1 (ZB
17), 2010-28 (ZC
7), 2010-35 (ZD
7), 2010-55-1 (ZE
3), 2010-42-2 (ZF
1), 2010-9 (ZG
1) (Liu Yongfeng, Chen Zhiyi, Liu Youzhou etc. 2001-2010 Jiangsu Province Pyricularia oryzae population mutation analysis. Jiangsu agricultural journal, 2010,26 (6): the 1233-1237) bacterial strain of 6 microspecies.Above bacterial strain provides by Institute of Plant Protection, academy of agricultural sciences, Jiangsu Province.
Blast resistance identification adopts method inoculation in seedling stage, and the rice material that need are identified after 3 days, rinses vernalization 1-2 days with clear water with antibiotic 402 seed soaking, is sowed in seedling-cultivation plate, broadcasts the 15-20 grain.Strains tested is transplanted on RCA (Semen Maydis powder 40g, rice straw 50g, agar 20g) substratum, cultivated 7 days under 25 ℃, after with black lamp, irradiating 72 hours, after Pyricularia oryzae produces spore, use again under aseptic washing, be made into the suspension of every visual field 30-40 spore under 10 * 10 power microscopes, at rice seedling 3 leaves during 1 heart stage, the isolation spray inoculation, every kind repeats 3 times, and moisturizing, after 7 days, is pressed the international uniform standard in shading shed, check incidence, the classification record.Blast resistance identification uses Suyunuo as susceptible check variety.
2,02428 pair of blast resistance identification result of wide affine kind
Rice seedlings rice blast grade scale:
0 grade does not have symptom (immunity)
The brown point (high resistance) of 1 grade very little
2 grades of brown scabs (disease-resistant) that diameter is 1mm
3 grades of diameters are the oval-shaped scab of being with of 2-3mm, central grey, edge brown (in anti-)
The oval scab of 4 grades of long 1-2cm, central grey, edge brown (susceptible)
5 grades form long and wide large oval scab, and scab develops into the later stage, edge brown (high sense)
02428 pair of wide affine kind blast resistance identification result in seedling stage is 0 grade, and 6 microspecies are to immunity, and contrast Suyunuo Resistance Identification result is 5 grades, is high sense.
According to known disease-resistant Recessive alleles
Pi21Sequence and disappearance Position Design primer thereof, wide affine kind 02428 and check variety Suyunuo are carried out to the pcr amplification of target gene Partial Fragment, as shown in Figure 4, the amplified production size of two kinds there are differences, the allelic genotype difference that these two kinds are entrained is described, and then the neomorph that wide affine kind 02428 is carried is described
Pi21tFor the high antimorph of rice blast wide spectrum.
Described allelotrope
Pi21tCan be used in rice blast broad-spectrum disease resistance breeding.
SEQUENCE LISTING
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aattttcaag gctaatcagc agtgttcctg ctcaattggg agcaaaacct ctgttaaaaa 180
gggtgtgttt gaatgaatat aattgaatat gaacgcagag gagtactgca tcgagaaggt 240
ggagtacgac gtgaagaaca acagggtgat cgtgcgcggg aagttcgacc cggagaagct 300
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gccgccgaag ccgtgcgggt gctcgcacgc cttcgtgtgc gtctgcaagc cggcgccgcc 720
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Met Gly Ile Leu Val Ile Leu Val Asp Leu Gln Cys Cys Arg Cys Asp
1 5 10 15
Ala Lys Ile Arg Lys Val Leu Gly Cys Leu Glu Glu Glu Tyr Cys Ile
20 25 30
Glu Lys Val Glu Tyr Asp Val Lys Asn Asn Arg Val Ile Val Arg Gly
35 40 45
Lys Phe Asp Pro Glu Lys Leu Cys Lys Lys Ile Trp Cys Lys Ala Gly
50 55 60
Lys Ile Ile Lys Glu Ile Leu Ile Val Asp Val Trp Pro Pro Pro Pro
65 70 75 80
Pro Cys Glu Lys Pro Pro Glu Asp Cys Lys Pro Lys Pro Cys His Cys
85 90 95
Cys Ser Cys Glu Lys Pro Lys Pro Lys Pro Lys Pro Cys His Cys Glu
100 105 110
Lys Pro Lys Pro Cys His Cys Glu Lys Pro Lys Pro Cys Glu Lys Pro
115 120 125
Pro Pro Glu Lys Pro Pro Pro Lys Pro Glu Cys Lys Leu Val Pro Tyr
130 135 140
Pro Tyr Pro Val Pro Tyr Pro Tyr Ala Gly Gln Trp Cys Cys Pro Lys
145 150 155 160
Pro Glu Pro Pro Lys Pro Pro Pro Glu Pro Pro Lys Glu Pro Glu Pro
165 170 175
Pro Lys Pro Cys Gly Cys Ser His Ala Phe Val Cys Val Cys Lys Pro
180 185 190
Ala Pro Pro Pro Pro Pro Pro Cys Gly Cys Ser Gly Gly His Gly Asn
195 200 205
Cys Gly Cys Gly Ile Arg Pro Trp Pro Pro Gln Val Trp Pro Pro Pro
210 215 220
Pro Val Cys Pro Pro Pro Pro Trp Cys Tyr Thr Glu Asp Asn Ala Asn
225 230 235 240
Ala Cys Ser Ile Met
245
Claims (5)
1. rice blast broad-spectrum disease resistance neomorph
Pi21t, it is characterized in that:
This equipotential gene
Pi21tTotal length is 874bp, and its sequence is:
ATGGGTATATTGGTCATCTTGGTGGACCTGCAATGCTGCCGCTGCGATGCCAAGATCAGGAAGGTCCTGGGCTGCCTTGAAGGTATAATAAATTCTGCCCGAATCGTCCATGTTTGATTGAATTTTCAAGGCTAATCAGCAGTGTTCCTGCTCAATTGGGAGCAAAACCTCTGTTAAAAAGGGTGTGTTTGAATGAATATAATTGAATATGAACGCAGAGGAGTACTGCATCGAGAAGGTGGAGTACGACGTGAAGAACAACAGGGTGATCGTGCGCGGGAAGTTCGACCCGGAGAAGCTGTGCAAGAAGATCTGGTGCAAGGCCGGCAAGATCATCAAGGAGATCCTCATCGTCGACGTCTGGCCGCCGCCGCCGCCGTGCGAGAAGCCTCCGGAGGACTGCAAGCCCAAGCCCTGCCATTGCTGCAGCTGCGAGAAGCCCAAGCCCAAGCCCAAGCCCTGCCACTGCGAGAAGCCCAAGCCCTGTCACTGCGAGAAGCCCAAGCCATGCGAGAAGCCGCCGCCGGAGAAGCCGCCGCCGAAGCCGGAGTGCAAGCTGGTGCCGTACCCTTACCCGGTGCCGTACCCGTACGCCGGGCAGTGGTGCTGCCCAAAGCCTGAGCCGCCGAAGCCGCCGCCGGAGCCACCGAAGGAGCCGGAGCCGCCGAAGCCGTGCGGGTGCTCGCACGCCTTCGTGTGCGTCTGCAAGCCGGCGCCGCCGCCGCCGCCGCCGTGCGGGTGCTCGGGGGGCCACGGGAACTGCGGCTGCGGCATCAGGCCGTGGCCGCCGCAGGTGTGGCCGCCGCCGCCCGTCTGCCCGCCGCCGCCGTGGTGCTACACCGAGGACAACGCCAACGCCTGCTCCATCATGTGA。
2. neomorph according to claim 1
Pi21t, it is characterized in that:
This equipotential gene
Pi21tFrom wide affine kind 02428,6 groups' of rice blast of Yangtze-Huaihe River Valley Major Epidemic Races is all shown as to high resistance, with respect to susceptible allelotrope
Pi21Have two place's sequence deletions, its deletion sequence is respectively: GCCGCTGCCGCAGCCGCCGCCGCCGTGCAA
And GAAGCCGCCGCCGTGCAAGCCGGAGGAGCCGCC.
3. the described neomorph of claim 1 or 2
Pi21tThe amino acid of deriving, its sequence is:
MGILVILVDLQCCRCDAKIRKVLGCLEEEYCIEKVEYDVKNNRVIVRGKFDPEKLCKKIWCKAGKIIKEILIVDVWPPPPPCEKPPEDCKPKPCHCCSCEKPKPKPKPCHCEKPKPCHCEKPKPCEKPPPEKPPPKPECKLVPYPYPVPYPYAGQWCCPKPEPPKPPPEPPKEPEPPKPCGCSHAFVCVCKPAPPPPPPCGCSGGHGNCGCGIRPWPPQVWPPPPVCPPPPWCYTEDNANACSIM-。
4. the described neomorph of claim 1 or 2
Pi21tApplication in the rice blast broad-spectrum disease resistance.
5. the described neomorph of claim 3
Pi21tThe application of amino acid in the rice blast broad-spectrum disease resistance of deriving.
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| CN108342502A (en) * | 2018-02-05 | 2018-07-31 | 西南大学 | It identifies the PCR primer pair of wheat ms1b recessiveness sterile genes and primer pair combines and its detection method |
| CN110656201A (en) * | 2019-11-23 | 2020-01-07 | 闽江学院 | Functional molecular marker of rice blast basic resistance gene pi21-2428 and application thereof |
| CN112575026A (en) * | 2020-12-19 | 2021-03-30 | 中国农业科学院植物保护研究所 | Application of combined knockout of rice susceptibility gene in creating broad-spectrum disease-resistant material |
| CN116286881A (en) * | 2023-03-28 | 2023-06-23 | 广西大学 | Sugarcane tip rot effector FsSCR1 gene and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN101906427A (en) * | 2010-06-25 | 2010-12-08 | 华南农业大学 | Rice blast resistance gene Pi1 and application thereof |
| CN102021176A (en) * | 2010-02-02 | 2011-04-20 | 华南农业大学 | Rice blast resistance gene Pik-h and application thereof |
| CN102094027A (en) * | 2010-12-08 | 2011-06-15 | 华南农业大学 | Rice blast resistance gene Pi7 and application thereof |
| CN102702337A (en) * | 2012-05-21 | 2012-10-03 | 中国科学院遗传与发育生物学研究所 | Rice blast disease-resisting protein, coding gene and application thereof |
-
2013
- 2013-08-12 CN CN2013103485279A patent/CN103409433A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN102021176A (en) * | 2010-02-02 | 2011-04-20 | 华南农业大学 | Rice blast resistance gene Pik-h and application thereof |
| CN101906427A (en) * | 2010-06-25 | 2010-12-08 | 华南农业大学 | Rice blast resistance gene Pi1 and application thereof |
| CN102094027A (en) * | 2010-12-08 | 2011-06-15 | 华南农业大学 | Rice blast resistance gene Pi7 and application thereof |
| CN102702337A (en) * | 2012-05-21 | 2012-10-03 | 中国科学院遗传与发育生物学研究所 | Rice blast disease-resisting protein, coding gene and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| SHUICHI FUKUOKA,ET AL: "Loss of Function of a Proline-Containing Protein Confers Durable Disease Resistance in Rice", 《SCIENCE》 * |
| 陆作楣: "论杂交稻育种的配合力选择", 《中国水稻科学》 * |
| 齐运田 等: "广亲和性品种02428与釉稻配组F1代优势分析", 《中国水稻科学》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108342502A (en) * | 2018-02-05 | 2018-07-31 | 西南大学 | It identifies the PCR primer pair of wheat ms1b recessiveness sterile genes and primer pair combines and its detection method |
| CN110656201A (en) * | 2019-11-23 | 2020-01-07 | 闽江学院 | Functional molecular marker of rice blast basic resistance gene pi21-2428 and application thereof |
| CN112575026A (en) * | 2020-12-19 | 2021-03-30 | 中国农业科学院植物保护研究所 | Application of combined knockout of rice susceptibility gene in creating broad-spectrum disease-resistant material |
| CN112575026B (en) * | 2020-12-19 | 2022-04-01 | 中国农业科学院植物保护研究所 | Application of combined knockout of rice susceptibility gene in creating broad-spectrum disease-resistant material |
| CN116286881A (en) * | 2023-03-28 | 2023-06-23 | 广西大学 | Sugarcane tip rot effector FsSCR1 gene and application thereof |
| CN116286881B (en) * | 2023-03-28 | 2023-09-05 | 广西大学 | Sugarcane tip rot effector FsSCR1 gene and application thereof |
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