CN1869232A - Paddy rice hybrid fertility gene and its application - Google Patents

Paddy rice hybrid fertility gene and its application Download PDF

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CN1869232A
CN1869232A CN 200610011976 CN200610011976A CN1869232A CN 1869232 A CN1869232 A CN 1869232A CN 200610011976 CN200610011976 CN 200610011976 CN 200610011976 A CN200610011976 A CN 200610011976A CN 1869232 A CN1869232 A CN 1869232A
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CN100457905C (en
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刘耀光
杨绍华
陈竹锋
庄楚雄
梅曼彤
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South China Agricultural University
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Abstract

The invention relates to the separation cloning and application of paddy hybrid gene Sb. It includes the paddy hybrid gene DNA sequence, the coding protein, and the DNA sequence carrier, and the host cell. The invention also relates to the method and application of the cultivation the compatible series by using the series and the molecule mark and the application to identify the gene.

Description

A kind of paddy rice hybrid fertility gene and application thereof
Technical field
The present invention relates to the genetically engineered field, be specifically related to a kind of separating clone and application in the paddy rice cross breeding breeding thereof of paddy rice hybrid fertility gene.
Background technology
Plant hybridizes by the hybridization between different varieties.Hybrid has advantage at aspects such as growth potential, adaptability, output, is called hybrid vigour.Utilize hybrid vigour can improve output, quality and the resistance of farm crop.The long-grained nonglutinous rice (indica) and japonica rice (japonica) inter subspecific hybrid of cultivated rice (Oryza sativa L.) have powerful hybrid vigour, and yield potential is very big.On the other hand,, can import cultivated rice, cultivate good new variety being present in some the useful proterties of wild-rice such as disease-resistant, pest-resistant, high yield etc. by hybridization of cultivated rice and wild-rice and breeding method such as backcross.But there is the low or sterile problem of fertility in the hybrid that Hybrids of Indica and Japonica and cultivated rice and wild-rice hybridization produce, is called hybrid dysgenesis (hybrid sterility).Described hybrid dysgenesis is one of isolated mechanism of reproduction that produces in the Oryza species atomization, belongs to behind the zygote reproduction and isolates (postzygotic isolation).The hybrid dysgenesis phenomenon has seriously hindered the utilization of rice heterosis and the useful gene of wild-rice.
Found that the locus more than 20 is relevant with paddy rice hybrid fertility; In these hybrid fertile gene seats, some is to cause megagamete sterile, as S5, and S7, S8, S9, S15, S17, some is to cause microgamete (pollen) sterile, as Sa, Sb, Sc, Sd, Se, (Kinoshita 1995, RiceGenet.Newsl., 12:9-15 for Sf; Ikehashi etc. 1988, Japan J.Breed.38:283-291; Ikehashi etc. 1996, Rice Genet III, pp404-408; Wan etc. 1996, Theor ApplGenet 92:183-190; Wang etc. 1998, Rice Genet Newsl 15:151-154; Zhang Guiquan etc. 1989, rice in China science 3:97-101; Zhang Guiquan etc. 1993, Acta Genetica Sinica 20:222-228; Zhang Guiquan etc. 1993, Acta Genetica Sinica 20:541-551; Zhang Guiquan etc. 1994, Acta Genetica Sinica 21:34-41).These genes can have variation differentiation in various degree in different varieties or strain or wild-rice material, produce multiple multiple allelomorphos.According to the paddy rice subspecies type of carrying these multiple allelomorphoss and to the effect of hybrid fertile, they are divided into indica type (S i), round-grained rice type (S j), affinity type (S n) or other transitional type.Described gene symbol S represents hybrid sterility (Sterility), S5, and Sa, Sb etc. refer to specific hybrid fertile gene seat, i, j, n represent the multiple allelomorphos of indica type (indica), round-grained rice type (japonica) and neutral type (neutral) respectively, as Sb i, Sb j, or Sb nNeutral type is also referred to as affinity type (compatibility) or wide affinity type (wide compatibility).
In paddy rice hybrid, not affine mutual work often takes place in the bigger hybrid fertile gene of differentiation degree under heterozygous state, cause carrying a certain allelic microgamete (or megagamete) grow undesired.This phenomenon is also referred to as the allelotrope selectivity of hybrid and kills the gamete effect.But these genes under homozygotic state (as the parent of isozygotying) do not influence the normal development of gamete.The genetic behavior of most of hybrid dysgenesis locus meets the pattern (Ikehashi etc. 1986, IRRI (ed) Rice Genetics:pp119-130) of " single seat position sporinite-gametophyte is done mutually ".For example, with carrying Sb respectively jSb jAnd Sb iSb iGenotypic paddy rice japonica rice and near isogenic line hybridization, the genotype of the first generation of hybrid of generation (F1) is Sb iSb jIn the microgamete of its generation, carry Sb iAllelicly normally can educate, and carry Sb jAllelic most of microgamete heteroplasia (sterile) causes F2 for carrying Sb in the colony jSb jThe ratio of genotypic plant is seriously on the low side, i.e. substantial deviation Sb iSb i: Sb iSb j: Sb jSb j=1: 2: 1 law of segregation.But, with carrying affinity type allelotrope Sb nSb nThe parent with carry Sb iSb iOr Sb jSb j(genotype of F1 is Sb in allelic parental rice hybridization nSb iOr Sb nSb j), F2 meets 1: 2: 1 law of segregation for 3 kinds of genotypic separation in the colony, and promptly the allelotrope of this heterozygosis does not produce selectivity and kills gamete effect, Sb nTo indica type and round-grained rice type allelotrope all is affine.
The heterozygosis S at different seats iS jGene can produce selectivity respectively and kill the gamete effect, therefore has the heterozygosis S at a plurality of seats in hybrid iS jCan produce the effect that adds up during gene, make hybrid dysgenesis even more serious.Otherwise,, just can improve the fertility of paddy rice hybrid if make hybrid strain carry affinity type allelotrope or phase isoallele at S locus as much as possible.For example, use wide affine system (kind) to be parent and long-grained nonglutinous rice or japonica rice hybridization in the present paddy rice cross breeding breeding usually, the fertility of its hybrid is improved to some extent.Described wide affine strain (kind) is meant at a plurality of hybrid dysgenesis locus carries the allelic strain of affinity type (kind).But available wide affine strain (kind) is less, and majority belongs to the intermediate type of the non-round-grained rice of non-Xian, and hybrid vigour is strong inadequately, and its economical character can not satisfy the diversified requirement of modern rice breeding.
Reported at present the chromosomal localization of some paddy rice hybrid fertility genes, but the separated clone's of paddy rice hybrid fertility gene report was not arranged before the present patent application proposes as yet, the molecular basis of hybrid dysgenesis or affine (can educate) is not clear.Sb, or be named as f5, be one of gene that influences rice hybrid pollen fertility, it has been located in the 5th karyomit(e) (Chuxiong, the village etc. 2002, Acta Genetica Sinica 29:700-705; Li Wentao etc. 2002, Botany Gazette 44:463-467; Li Wentao etc. 2006, Science Bulletin 51:404-408; Wang etc. 2006, Theor.Appl.Genet.112:382-387).In order in breeding of hybridized rice, to overcome the hybrid sterility problem effectively, the present invention has obtained a plurality of allelotrope of paddy rice hybrid fertility gene Sb by map based cloning (map-based cloning) technology, comprise affinity type allelotrope, verified the function of this gene by transgenic experiments, and proposed by the crossbreed method of the affine system of fertility of genetic engineering technique or hybridization-back cross breeding.
Summary of the invention
(1) goal of the invention
The objective of the invention is separating clone paddy rice hybrid fertility gene and the dna fragmentation that comprises the promotor of regulating and control this gene.
Second purpose of the present invention provides the coded protein of above-mentioned hybrid fertile gene.
The 3rd purpose of the present invention provides the application of above-mentioned hybrid fertile gene in the affine system of the fertility that crossbreeds.
A further object of the present invention provides molecule marker or its close linkage mark and the application in the paddy rice cross breeding breeding thereof that above-mentioned hybrid fertile gene produces.
(2) technical scheme
The present invention relates to separate and use a kind of dna fragmentation that comprises hybrid fertile gene Sb, this fragment is given between rice varieties (being) or the microgamete of the hybrid that paddy rice and wild-rice intermolecular hybrid produce has normal fertility.
The present invention utilizes the map based cloning method to clone the Sb gene, and its technological step is as follows:
1, the chain location of the screening of polymorphic molecular marker and Sb gene
At present Sb is positioned approximate region (Chuxiong, the village etc. 2002, Acta Genetica Sinica 29:700-705) on paddy rice the 5th karyomit(e).According to the data (http://www.ncbi.nlm.nih.gov) in disclosed Public Rice Genome Sequence Data storehouse, in this regional development a plurality of molecule markers.Pass through linkage analysis, find 6 and Sb close linkage or common isolating polymorphism mark, and the interval of Sb Fine Mapping at a 5kb.
2, dna sequence analysis
According to the data (http://www.ncbi.nlm.nih.gov) in disclosed Public Rice Genome Sequence Data storehouse, in localized 5kb interval, has only the gene of 1 prediction.
3, the functional verification of Sb gene
With PCR method from 1 affine strain WS59 amplification affinity type allelotrope Sb n, it comprises promoter region, coding region and downstream sequence, makes up the Plant Transformation expression vector.Transform japonica rice T65 with the soil agrobacterium tumefaciens-mediated transformation, again with near isogenic line E2 hybridization, the F1 that hybridizes, its genotype is Sb iSb j/ Sb n-, Sb wherein iSb jBe the endogenous Sb genotype of hybrid, Sb nBe the transgenosis that imports.Import Sb nThe fertile flower powder rate of hybrid plant improve, among its self progeny F2, contain Sb jSb jThe ratio of genotypic plant improves greatly, shows most of Sb jThe fertility restorer of type microgamete is normal.These results have proved Sb nAllelotrope has the biological function that overcomes hybrid dysgenesis.
4, the sequencing comparative analysis of Sb multiple allelomorphos
Amplify the Sb gene fragment with round pcr from a plurality of indica type, round-grained rice types and affinity type kind (being), measure its sequence after being cloned into plasmid vector.Wherein the nucleotide sequence of 3 affinity type genes is shown in sequence table SEQ ID NO:1, SEQ ID NO:3 and SEQ ID NO:5.They are encoded respectively and have the amino acid polypeptide sequence shown in SEQ ID NO:2, SEQ ID NO:4 and the SEQ ID NO:6, and its biological function is the pollen fertility of controlling plant hybrid.These multiple allelomorphoss are all encoded and are contained 662 amino acid whose protein, do not have the conserved domain of known functional protein matter, do not find the gene of homology with it and identical function.The variation of several bases is arranged in the coding region between the described multiple allelomorphos, and cause several amino acid whose variations (table 2).Not affine or affine mutual work takes place in the microgamete that these variations have caused containing the hybrid of various combination heterozygous genes.
According to Sb gene order information provided by the invention, those skilled in the art can easily obtain identical with Sb or are equal to the gene of Sb by following or other genetic engineering technique, and perhaps its function is equivalent to the subfragment of sequence shown in SEQ ID NO:1 or SEQ ID NO:3 or the SEQ ID NO:5: genomic library or cDNA library that (1) classifies the probe Screening of Rice as with Sb gene or homologous fragment or its other adjacent genome sequence obtain; (2) design oligonucleotides primer (containing degenerated primers) increases from the genome of cultivated rice or wild-rice or cDNA through polymerase chain reaction (PCR) and to obtain; (3) modifying transformation with gene engineering method on the basis of Sb gene order obtains; (5) method with chemosynthesis obtains.The described gene that is equal to or is equivalent to Sb, be meant that its encoded protein matter sequence compares the difference of having only one or several amino-acid residue with the amino acid residue sequence shown in SEQ ID NO:2, SEQID NO:4 or the SEQ ID NO:6, comprise replacement, the disappearance of amino-acid residue or add.And these difference do not cause its function and the function of Sb gene provided by the invention that in essence different are arranged.Especially, shown in embodiment provided by the invention, the Sb gene in Different Rice Varieties or strain and the different wild seed rice can exist variation in various degree, i.e. multiple allelomorphos.These multiple allelomorphoss can easily obtain with aforesaid method according to information disclosed by the invention.
The present invention includes such protein sequence, the aminoacid sequence shown in itself and SEQ ID NO:2, SEQ ID NO:4 or the SEQ ID NO:6 has about at least 70% or higher homology, and fertility that can the controlling plant hybrid.
The present invention also comprises the host cell that contains described nucleotide sequence or its segmental cloning vector or expression vector, contain described carrier, contains described nucleotide sequence or its segmental plant transformed cell and transgenic plant.
Hybrid fertile gene provided by the invention has important use and is worth.One of application is the affine system of fertility that crossbreeds.Therefore the present invention also provides the method for cultivating the affine system of paddy rice hybrid fertility by genetic engineering technique or the hybridization-method of backcrossing, and comprise the affine system of paddy rice hybrid fertility and other rice varieties or the incross of further using acquisition, cultivate the affine system of hybrid fertile that makes new advances.Of the present invention gene constructed in plant conversion carrier, can do suitably to modify to described gene or its regulating and controlling sequence, also can before its transcription initiation codon, replace the original promotor of described gene, make it to reach expression effect equal or that strengthen with other promotor.
The Another application of hybrid fertile gene provided by the invention is to produce specific molecule marker or its close linkage mark according to described gene order information, includes but not limited to SNP (mononucleotide polymorphic), InDel (it is polymorphic to insert disappearance), RFLP (restriction enzyme length is polymorphic), CAP (the cutting amplified fragments is polymorphic).Can identify the genotype of cultivated rice or wild-rice or their filial generation colony plant with these marks, use the molecular marker assisted selection breeding, thereby improve the efficiency of selection of breeding.
(3) beneficial effect
According to gene provided by the invention, can be with any plant conversion carrier or method for transformation with described affinity type (Sb n) allelotrope importing japonica rice or rice variety or strain, the affine system of hybrid fertile of acquisition energy expressing said gene; Or be parent and other rice varieties or incross with the transformed plant that obtains, the affine system of hybrid fertile that cultivation makes new advances.Also can be with carrying Sb nAllelic cultivated rice or wild-rice material are donor parents, and other japonica rice or rice variety (being) are recurrent parent, hybridize and backcross, and utilize the specific molecular marker that produces according to the present invention to advance screening, cultivate with Sb nSubstitute original Sb iOr Sb jThe affine system of new hybrid fertile.Obtain the affine system of hybrid fertile and other japonica rice or long-grained nonglutinous rice or wild-rice parent hybridization with method of the present invention, can produce the normal hybrid of fertility, help new breed breeding or produce cross-fertilize seed.On the other hand, produce the genotype that molecule marker can be identified paddy rice or wild-rice or filial generation plant, be used for the molecular marker assisted selection breeding, thereby improve the efficiency of selection of breeding with the present invention
Description of drawings
The hereditary pattern of Fig. 1, paddy rice Sb gene and genotype thereof are to the effect of pollen fertility.(A) japonica rice T65 (Sb jSb j) the normal fertile flower powder that produces; (B) near isogenic line E2 (Sb iSb i) the normal fertile flower powder that produces; (C) the hybrid F of T65 and E2 hybridization 1(Sb iSb j) pollen that produces, wherein about 50% pollen sterility (dyeing of arrow indication is more shallow, the pollen that diameter is less); (D-F) F of T65 * E2 hybrid 2In the plant, ratio of different genotype plant and pollen fertility thereof.According to the evaluation with linked marker, F 2Sb in the colony jb jThe genotype plant is seriously on the low side, and F is described 1In the pollen that plant produces, most of Sb jGenotypic pollen sterility;
The chromosome linkage location of Fig. 2, paddy rice Sb gene, clone, functional analysis and with the crossbreed synoptic diagram of the affine system, method of fertility of transgenic technology;
Fig. 3, with round pcr from different rice material amplification Sb gene fragment.M is a molecular weight marker;
Fig. 4, the expression of usefulness reverse transcription PCR technology for detection Sb gene in young fringe;
The structure of Fig. 5, (A) Sb gene coding region, box indicating exon, the start-stop position in numeral exons coding district (contain and end password).(B) Sb of WS59 nThe allelotrope amino acid sequence coded;
Fig. 6, from the Sb that contains of the affine WS59 of being amplification nAllelic dna fragmentation is cloned into plant conversion carrier pCAMBIA1300.1-3 is the electrophorogram with 3 clones of BamHI/HindIII double digestion.M is a molecular weight marker;
Fig. 7, with affinity type gene Sb nImport T65 and obtain the T0 transformant, hybridize the Sb that produces with E2 again iSb j/--pollen (the arrow indication is sterile pollen) and the Sb of genotype hybrid (A) iSb j/ Sb nThe pollen of-genotype hybrid (B-D) (major part normally can be educated);
Fig. 8, usefulness single nucleotide polymorphism (SNP) mark 07-81 identify the genotype of parent and hybrid plant (1-6).
Embodiment
Following examples further specify content of the present invention.
Embodiment 1: the screening of polymorphic molecular marker and the Fine Mapping of Sb
Known Sb is positioned at the approximate region (Chuxiong, the village etc. 2002, Acta Genetica Sinica 29:700-705) on paddy rice the 5th karyomit(e).The present invention further clones the Sb gene with map based cloning method as shown in Figure 2.At first use in the japonica rice variety platform 65 (to be called for short T65, to have genotype Sb jSb j) and its near isogenic line E2 (Sb iSb i) hybridization, set up a F 2For segregating population.The near isogenic line E2 that the present invention is used is except Sb iOutside the allelotrope, the nuclear gene group background strain similar to T65, so the male infertility of their hybrid only is subjected to Sb iSb jHeterozygous genes Make effect mutuallyInfluence.Use 1%I 2-KI solution-dyed is identified the form of pollen, has determined the F of strain more than 6000 2The pollen fertility phenotype of plant promptly can be educated phenotype or half sterile phenotype (Fig. 1), entirely with these F 2Plant is used for the Fine Mapping of Sb.
According to the data (http://www.ncbi.nlm.nih.gov) in disclosed Public Rice Genome Sequence Data storehouse, in this regional development a plurality of polymorphic molecular markers, as shown in table 1.
The recombination event that molecule marker and target are recovered between the locus is many more, represent that both genetic distances are far away more, otherwise its genetic distance is near more.Utilize the method and the above-mentioned F of strain more than 6000 of this genetic mapping 2Plant has made up the genetic linkage map (Fig. 2) of a Sb gene.It is to have only 1 recombination event between 07-78 and 07-83 and the Sb respectively that 2 marks are arranged among the figure, and mark 07-81 and Sb gene to be zero reorganization promptly be divided into from.Physical distance between 07-78 and the 07-83 is about 5kb.Described locating area be positioned at disclosed paddy rice bacterial artificial chromosome (bacterial artificial chromosome, BAC) P0008A07 ( Http:// www.ncbi.nlm.nih.gov) about 78-83kb place.
Table 1 is with the Sb close linkage or be total to isolating polymorphic molecular marker
Molecule marker The primer title Primer sequence
07-55 55F 55R 5′-GCAGTTACCGGCAACGAGG-3′ 5′-GACTGACCCCTACCACCAC-3′
07-78 78F 78R 78R-i 78R-j 5′-GGGAGTACGTGGCATGTITG-3’ 5′-GACATTCACTCATATCGGCAACG-3’ 5′-GGTCTTCGTIGGTAACCTCCA-3’ 5′-GGTCTTCGTTGGTAACCTCCG-3’
07-81 81F 81R 81R-i: 81R-j 5′-CTCGGATACAATGATGAAGCAG-3’ 5′-GGAAAGCTACTGTACCAACAG-3’ 5′-CTTGATTGATTCCATTGACAA-3’ 5′-CTTGATTGATTCCATTGACAT-3’
07-83 83F 83R 83R-i 83R-j 5′-CTCAGTTTGTTGTGGAGCACGAC-3’ 5′-ACCAGGTACAATGTGCAACCAAC-3’ 5′-CACTACATATCATAATAA-3’ 5′-CACTACATATCATAATAC-3’
07-86 86F 86R 86R-i 86R-j 5′-GCAGACAAAGCAGATGGATG-3’ 5′-CCCACAGCAAGCCTTATGAG-3’ 5′-GTCATCACATCTCAAACAATGA-3’ 5′-GTCATCACATCTCAAACAATGG-3’
07-130 130F 130R 5′-TTCGGGATGAAAGTGGTGAG-3’ 5′-GAATGTATCCGTITCCAGC-3’
07-55 in the table 1 and 07-130 are insertion/disappearance (InDel) mark; All the other are single nucleotide polymorphism (SNP) mark.The SNP detection method of PCR-based is: add the 1ul paddy DNA in 20ul PCR reaction solution, with the primer " F " shown in the table 1 and " R " amplification 33 circulations (94 ℃ of 30s, 63 ℃ of 1min, 72 ℃ of 1min).Get the template that 0.5ul PCR product is the 2nd PCR then, each sample 2 PCR reaction of preparation (20ul), promptly use primer " F " and " R-i " to be the specific reaction of raq gene type, with primer " F " and " R-j " is the genotypic specific reaction of T65,8-10 circulation (94 ℃ of 30s increase, 58 ℃ of 1min, 72 ℃ of 1min).Detect the PCR product with 1% sepharose.
The clone of embodiment 2:Sb and sequential analysis
According to the disclosed paddy rice of GenBank (japonica rice) genomic dna sequence and RiceGAAS software ( Http:// ricegaas.dna.affrc.go.jp) analyze, in localized 5kb zone, only there is the gene of 1 prediction.1 662 amino acid whose protein of this genes encoding does not have the conserved domain of known functional protein matter.The database contrast ( Http:// www.ncbi.nlm.nih.gov, blastn, blastp) do not find the gene of homology with it and identical function therefore to the present invention is to provide a new functional gene.
From 3 affine systems (WS59, Dular, 02428), near isogenic line (E2), rice variety (93-11), and japonica rice variety (T65, Japan is fine) extracts total DNA.DNA of plants trace extraction method is: get the about 0.2g of blade, put into the centrifuge tube of 2ml, add liquid nitrogen, with Phillips screwdriver blade is stampped powdered, add 0.8ml extracting solution (0.5MNaCl, 0.1M Tris-HCl (pH8.0), 0.05M EDTA, 1.25%SDS) shake up, place 30min for 65 ℃.Add 0.8ml chloroform/primary isoamyl alcohol/ethanol (76: 4: 20), concussion 5min, the centrifugal 5min of 12000rpm.Take out supernatant liquor, add 2 times of volume ethanol deposit D NA, the centrifugal 5min of 12000rpm removes supernatant.With 0.5ml 70% ethanol rinsing precipitation, air-dry, be dissolved in 0.1mlTE solution.Use primer 5 '-CATAC GGATCCCATACATACCAATACAAACC-3 ' (underscore is BamHI) and 5 '-GTTATG AAGCTTGACAATGGACCTGACATG-3 ' (underscore is HindIII) amplifies Sb gene fragment (Fig. 3) with round pcr from these paddy DNAs.High-fidelity LA-Taq enzyme (Bao Bio-Engineering Company), 30 circulations of increasing (94 ℃ of 30s, 62 ℃ of 1min, 72 ℃ of 5min) are used in this PCR reaction.After cutting with the BamHI/HindIII enzyme, connect and be cloned into plasmid vector pUC18.Through sequencing analysis, find to have in the coding region between these multiple allelomorphoss the difference of base, and cause 2-12 amino acid whose difference (table 2).
To the expression (Fig. 4) of Sb gene in young fringe tissue, and confirmed the coding region structure of this gene (Fig. 5 A) with primer 5 '-CCGTCTACGACCGGCGGT-3 ' and 5 '-CCTAAACAGCACACTTACAGA-3 ' reverse transcription PCR technology for detection by measuring its cDNA sequence.Affine is the Sb of WS59 nThe allelotrope amino acid sequence coded is shown in Fig. 5 B.
The allelic aminoacid sequence difference of the Sb of many rice varieties of table 2 (being).
Strain Amino acid position
225 227 228 231 290 351 382 386 403 491 493 505 547 605 625 659
WS59 Dular 02428 E2 93-11 T65 Japan is fine K K K E K K K S S S N N S S H H H R R H H L S S L S S S F F S F F F F S S S C C S S A T A A A A A A V E E E E E S S S Y S S S N N N Y Y N N L L M L L M M S Y Y Y Y Y Y G R G G G G G I I V V V V V T T A T T T T L R L L L L L
Embodiment 3: genetic transformation is identified the function and the application thereof of target gene
To be cloned into double base conversion carrier pCAMBIA1300 (Australian CAMBIA company) from the dna fragmentation (Fig. 3) of the affine WS59 of being amplification, obtain called after pCAMBIA1300-Sb nExpression vector plasmid (Fig. 6).This fragment comprises Sb nCoding region and upstream transcription control region and downstream sequence.With electrization this expression vector is imported soil agrobacterium tumefaciens (Agrobacterium tumefaciens) bacterial strain EHA105 commonly used.Transform T65 by agrobacterium tumefaciens-mediated transformation (Hiei etc., Plant are J.6:271-282), obtain T0 for transfer-gen plant (Sb jSb j/ Sb n-), Sb wherein jSb jBe native gene, Sb nFor transforming the gene that imports, Sb n-be expressed as the transgenosis hemizygote.With transformed plant and E2 hybridization, the hybrid F of generation 1Hybrid has 2 kinds of genotype, i.e. Sb iSb j/--(there is not Sb n) and Sb iSb j/ Sb n-(there is Sb nHemizygote).
Hybrid F 1Get pollen during heading with 1% potassiumiodide (I 2-KI) dyeing microscopic examination.At Sb iSb j/--genotypic hybrid, its fertile flower powder rate is about 50% (Fig. 7 A).Be divided into 3 kinds of genotypic numbers analyzing 434 F2 plant from mark 07-81 with Sb, find Sb iSb i, Sb iSb j, and Sb jSb jThe genotype plant is respectively 205 strains (47.2%), and 218 strains (50.2%) and 11 strains (2.5%) show most Sb jThe type microgamete is sterile, Sb jAllelotrope mainly is delivered to the offspring by megagamete.Import Sb and contain nHybrid F 1(Sb iSb j/ Sb n-) fertile flower powder rate bring up to about 80% (Fig. 7 B-D).Analyze 507 F 23 kinds of genotypic numbers of plant, Sb iSb i, Sb iSb j, and Sb jSb jThe genotype individuality is respectively 158 strains (31.1%), 248 strains (48.9%) and 101 strains (20.0%), Sb jSb jThe individual ratio of genotype increases substantially, and shows Sb jThe fertility of type microgamete obtains to recover.These results have proved Sb nAllelotrope has the Sb that recovers hybrid jGenotype is recovered the biological function of fertility.
With affinity type allelotrope Sb nImport in the japonica rice, and contain indica type allelotrope Sb iIncross, can obtain the normal hybrid of male fertile.This transgenosis evidence, Sb provided by the invention nGene is the paddy rice hybrid fertility compatibility gene that function is arranged, and can cultivate the affine system of hybridization fertility by transgenosis and selfing purifying.According to the present invention, similarly also can be with Sb nGene imports in the long-grained nonglutinous rice, obtains the affine system of hybrid fertile.Also can be with carrying Sb nAllelic cultivated rice or wild-rice material are donor parents, and other japonica rice or rice variety (being) are recurrent parent, hybridize and backcross, and utilize the specific molecular marker that produces according to the present invention to advance screening, cultivate with Sb nSubstitute original Sb iOr Sb jThe affine system of new hybrid fertile.The affine system of hybrid fertile and other japonica rice or rice variety (being) or wild-rice hybridization and selection with method of the present invention obtains can produce the higher hybrid of fertility, help the new kind of seed selection or produce cross-fertilize seed.
The application of embodiment 4:Sb gene order in the molecular marker assisted selection breeding
Utilize sequence information provided by the invention can produce the allele specific oligonucleotide molecule marker of Sb, be used to identify the plant that has the range gene type on the Sb site, can in the molecular marker assisted selection breeding process, be applied.For example, the mark 07-81 shown in the table 1 results from the Sb gene order, can be used for identifying the genotype (Fig. 8) of each hybrid plant.
Sequence table
<110〉Agricultural University Of South China
<120〉a kind of paddy rice hybrid fertility gene and application thereof
<130>
<160>6
<170>PatentIn version 3.3
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<211>4200
<212>DNA
<213〉Oryza rice strain WS59
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<221>CDS
<222>(1292)..(1861)
<220>
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<222>(1996)..(3004)
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aaaatcatcc tgttgagggt gttgtagcaa attttatctt tttttcatac atgcaatttc 60
ataaattttt ataaactgta atttttgtaa tgtttaggaa ggagtagtct ctttgtgttt 120
tcttttcaag agtaaactgc acaaacagtc cctaaacttg tgatggtgta tcacataggt 180
ccctaaactc tcaaaatgta ttttttgtgt tcccgaactt gtctagggtg tcgtataggt 240
ctaaacaagc gctaatccga cccatgtgtt gacttgacat gtcacaatgg tttctacatg 300
ttggtgggac ccacatgtca atcacataaa aaaattgaag gatagaggag acaaaaatgt 360
ttttttttaa ttttgaaaaa aaaagagagg agagagagag atgagaaaaa gatgttttta 420
aatttttatt ctatgtgtgt ttgataagtg ggtcccacta acacgtaggt gccactatgg 480
tattgtcatg tcagtgtaca aacctacatg acagtcaaga taagtttgga gacaagaaga 540
aaagaatttt aagagtttag ggatatagat gatatactac gcatgtgcac ttcactcgtt 600
ttgcaatgtt gcaattgatt ttccttttta tgtttcacgt gctggggtct tggtggtgtt 660
gcaacaagta ttgttgactg ttgcatctgt tagaaatatg catatgcatg tttcatcgag 720
tgaggaacta tttgaaacag ctagcggggt ttgaaatttc gtttcaaaat ttccgaaatt 780
tcagtaatct catatatagc ccgaaatttt cggttttttg caaatcttta tgaatttggt 840
caaattttat tcaaattcat tcaaaatcag tcaaaatgtt aaataatttc ggccaaaaaa 900
aaaagaaatt cccgaaattt ctgtgccccc ggcagaaata gcaatttcga aatttaaaac 960
tgtgagctag ccaaggaaat taattaagca caaacagcgc agcttagcga agtgagtgag 1020
tgaaggagtg cgcgcatcgc ttcgcctgga gagaccacct tcttctccaa tccccagttt 1080
tctccccttg ctgcttttgc ccgcccccct ctccccccca aaaaaaacca aaaaaaaaaa 1140
aagaagaaga agaagaagaa gaagaaagga agaagaaaac ctagctagtg tgaatcgcga 1200
gcacttaccg atgcttcgaa gggcgacggc tgcggtggcc ggccgccgtc tacgaccggc 1260
ggtgggcctc tcgccgtcgg ccccggcgag a atg gtg atg ggt tgc acc cac 1312
Met Val Met Gly Cys Thr His
1 5
cag cag ctc gcc gac cca tgg gca tgg gca gcc cgc gag gaa ggt ggc 1360
Gln Gln Leu Ala Asp Pro Trp Ala Trp Ala Ala Arg Glu Glu Gly Gly
10 15 20
cgt gtc gcc ggc agc tcc ctg cca cgg cgg ccg acc cct acc acc gcc 1408
Arg Val Ala Gly Ser Ser Leu Pro Arg Arg Pro Thr Pro Thr Thr Ala
25 30 35
atc ggc cgg gtc gcc gcc ggg cgc cgc cca cct gca ccg gca act ggc 1456
Ile Gly Arg Val Ala Ala Gly Arg Arg Pro Pro Ala Pro Ala Thr Gly
40 45 50 55
cca ccg cag ggg acg ccc gcc atg tcg ctg gca cca cca cca cca aag 1504
Pro Pro Gln Gly Thr Pro Ala Met Ser Leu Ala Pro Pro Pro Pro Lys
60 65 70
gcc tcg cct tcc gcc gcc cgc ctg cag aag ccg gca gct tcc tgc cac 1552
Ala Ser Pro Ser Ala Ala Arg Leu Gln Lys Pro Ala Ala Ser Cys His
75 80 85
ggc ggc caa cca aag gcc tcg cct tgg gcc ggg tcg ccg ccg gga gca 1600
Gly Gly Gln Pro Lys Ala Ser Pro Trp Ala Gly Ser Pro Pro Gly Ala
90 95 100
gcc acc cac ctg cag cag ccg tcg cgg atg aag ctc tct ccg tcg ccg 1648
Ala Thr His Leu Gln Gln Pro Ser Arg Met Lys Leu Ser Pro Ser Pro
105 110 115
gca cct ggc gcc caa cac ctg cca ccg gca aag ccc tcg cct ttg gcc 1696
Ala Pro Gly Ala Gln His Leu Pro Pro Ala Lys Pro Ser Pro Leu Ala
120 125 130 135
ggg tcg ccg ccg gga gca gcc acc cac ctg cag cag ccg gcg gcg agg 1744
Gly Ser Pro Pro Gly Ala Ala Thr His Leu Gln Gln Pro Ala Ala Arg
140 145 150
atg ctg gcc tcg tcg ccg gcg aag agg gtg ctc tca gtc ttc tca cca 1792
Met Leu Ala Ser Ser Pro Ala Lys Arg Val Leu Ser Val Phe Ser Pro
155 160 165
tca gcg gcc ggg tcg ctg cgg agg gaa cgt aac cat ctg ccg cca ttg 1840
Ser Ala Ala Gly Ser Leu Arg Arg Glu Arg Asn His Leu Pro Pro Leu
170 175 180
gcc agc tgc ttc tgc ttg cgg gtgggtaact ccaaattcca aatttaggtc 1891
Ala Ser Cys Phe Cys Leu Arg
185 190
caattaaagg acccccaatt tcctcgcttg tagctagtct tatcttattt gggtcatgta 1951
ttcattgact gatgatttgg gtcgtgtata tattgtgttt gcag cgg ctg atg agt 2007
Arg Leu Met Ser
acc gga cta cct caa att cca agt ccc cca gca aca ctg atg aga cct 2055
Thr Gly Leu Pro Gln Ile Pro Ser Pro Pro Ala Thr Leu Met Arg Pro
195 200 205 210
tgg tgt aat gag gat gag cag gtg cct caa cta acc cct ttt aag atc 2103
Trp Cys Asn Glu Asp Glu Gln Val Pro Gln Leu Thr Pro Phe Lys Ile
215 220 225
agt cat ggt gtc ttg gat aca atg atg aag cag tat gag gaa gac tac 2151
Ser His Gly Val Leu Asp Thr Met Met Lys Gln Tyr Glu Glu Asp Tyr
230 235 240
aag tat ctt gtg gag gag ggg ctg gat ctt gaa gag atg aag aaa ctt 2199
Lys Tyr Leu Val Glu Glu Gly Leu Asp Leu Glu Glu Met Lys Lys Leu
245 250 255
tac cat gag ctg gtg cgt ccc gtt tta gtt gca agc tgt ctg att ctt 2247
Tyr His Glu Leu Val Arg Pro Val Leu Val Ala Ser Cys Leu Ile Leu
260 265 270
gaa cat cac att gtc agt cct gaa acc aaa gcg cta gat cag aaa ttc 2295
Glu His His Ile Val Ser Pro Glu Thr Lys Ala Leu Asp Gln Lys Phe
275 280 285 290
aaa gac aag ttc ggg aaa ccc cca atc tat ctc tca gct ctt gat ata 2343
Lys Asp Lys Phe Gly Lys Pro Pro Ile Tyr Leu Ser Ala Leu Asp Ile
295 300 305
atg aag acc ttg att gct atg gag ccg tcg aga tgg agt tac ctt atg 2391
Met Lys Thr Leu Ile Ala Met Glu Pro Ser Arg Trp Ser Tyr Leu Met
310 315 320
gat ttc tta aga gag aaa cgt ata ttg gtc act gag aat tgg gaa gtt 2439
Asp Phe Leu Arg Glu Lys Arg Ile Leu Val Thr Glu Asn Trp Glu Val
325 330 335
tca ctg ggt tcc atc gag tgc atc atg cac gac gac agc att agt ttt 2487
Ser Leu Gly Ser Ile Glu Cys Ile Met His Asp Asp Ser Ile Ser Phe
340 345 350
atg atg aat ggc cgg gtc gtt ttc aaa ggg caa tct gat ggt agg tat 2535
Met Met Asn Gly Arg Val Val Phe Lys Gly Gln Ser Asp Gly Arg Tyr
355 360 365 370
aac cat ggg act ggt aca gcc gat ctt tgc aca gcg att act ggt gca 2583
Asn His Gly Thr Gly Thr Ala Asp Leu Cys Thr Ala Ile Thr Gly Ala
375 380 385
gct aca gat atc gtt gat tgt gcc atc tac cga gag gtc cct tgc aat 2631
Ala Thr Asp Ile Val Asp Cys Ala Ile Tyr Arg Glu Val Pro Cys Asn
390 395 400
tct cca acg cac gct gag ttg ctt gga atg tat gtt cta gaa aga agg 2679
Ser Pro Thr His Ala Glu Leu Leu Gly Met Tyr Val Leu Glu Arg Arg
405 410 415
gca ata tcg ctg aaa ata ctg ttg ttt gat gtg aaa acc gac aac gct 2727
Ala Ile Ser Leu Lys Ile Leu Leu Phe Asp Val Lys Thr Asp Asn Ala
420 425 430
ttt gtc agt gaa aca gtc aga gat atg ttt cct att act cca aac acc 2775
Phe Val Ser Glu Thr Val Arg Asp Met Phe Pro Ile Thr Pro Asn Thr
435 440 445 450
act gag aag gat ctg tgc caa gtg ttg aga agt atg aaa gtc tat ttc 2823
Thr Glu Lys Asp Leu Cys Gln Val Leu Arg Ser Met Lys Val Tyr Phe
455 460 465
gaa cac ttc aat tgt cgg tgt gaa cct cgg gag aaa ctt gag cta gtg 2871
Glu His Phe Asn Cys Arg Cys Glu Pro Arg Glu Lys Leu Glu Leu Val
470 475 480
gat agt tta atg aaa atg aag gat aat gag ttg aca atg gaa tca atc 2919
Asp Ser Leu Met Lys Met Lys Asp Asn Glu Leu Thr Met Glu Ser Ile
485 490 495
aag gat aaa tgg gct cat tcc ctt ctg aga ttg cct gtc ttc agg gct 2967
Lys Asp Lys Trp Ala His Ser Leu Leu Arg Leu Pro Val Phe Arg Ala
500 505 510
cat cag cca acc aag acc ata aga aag gac tat ata a gtaagtagtt 3014
His Gln Pro Thr Lys Thr Ile Arg Lys Asp Tyr Ile
515 520 525
aatcttctga gtacattgac tttgattttt ttttcttttt tcttttaacc catcggttta 3074
atttttatat tgcag ac aag gct cca act gtt ggt aca gta gct ttc cat 3124
Asn Lys Ala Pro Thr Val Gly Thr Val Ala Phe His
530 535
ggg caa tac aaa att gtc gca aag ggg gat aca gca aaa acg gag gct 3172
Gly Gln Tyr Lys Ile Val Ala Lys Gly Asp Thr Ala Lys Thr Glu Ala
540 545 550
gtt gtg aac ttg gtg cta tcg ctt cag cca atg gtt gct tca gtt ttc 3220
Val Val Asn Leu Val Leu Ser Leu Gln Pro Met Val Ala Ser Val Phe
555 560 565 570
gtg gga aat cac atc tca gca atc tca gtc aag gaa gag ata gaa aag 3268
Val Gly Asn His Ile Ser Ala Ile Ser Val Lys Glu Glu Ile Glu Lys
575 580 585
ttt gtc aag ggg tat tta ctg gac atc aag gag gct aag gga gca tat 3316
Phe Val Lys Gly Tyr Leu Leu Asp Ile Lys Glu Ala Lys Gly Ala Tyr
590 595 600
gtt gtt atc atg aaa ttt ata gaa act ccc aat gca ggt ggt aaa ata 3364
Val Val Ile Met Lys Phe Ile Glu Thr Pro Asn Ala Gly Gly Lys Ile
605 610 615
tta gat gtc ttg ttt aat act gaa gtc ggt gag gtg tac aag cag aca 3412
Leu Asp Val Leu Phe Asn Thr Glu Val Gly Glu Val Tyr Lys Gln Thr
620 625 630
gca aac tcg gtc tgc gtc aaa cta ctg gac aag aat gat gga act act 3460
Ala Asn Ser Val Cys Val Lys Leu Leu Asp Lys Asn Asp Gly Thr Thr
635 640 645 650
cca gag cta agt cca gca tgt ctt ctt att ttc ggt tgagtcacaa 3506
Pro Glu Leu Ser Pro Ala Cys Leu Leu Ile Phe Gly
655 660
agggtcaaac tagtagtttc ttggcatgtg ccttctgtaa gtgtgctgtt taggtggctg 3566
gagggccctt tttggttagt agtctagacc ctaacaacat gttattactt atgctgctgt 3626
tatgcaccct gtgcagcacg cttctgaatt gcaacttctt ttcaagctga atttatctgc 3686
gagtgttaac cctattttgt gatggtcata tatgtttatg gatggagctt caagctgggg 3746
catctgcagg gatctgagta ccagaactga agttctttgt gaaagaagtt gttttgcttg 3806
tagatggcaa tttccatgca gtctcatctg tacaaccccc aaaaagcatc attctgtttc 3866
agctacgagc tctgtgtttg tgcttttgtt tttcttcttg tttaccccag gatgggatgg 3926
caatggtagg gaatgctggt gtatctagac cgtgtttgtg cttttgtttt tcttcttgtt 3986
taccccagga tgggacggca atggtaggga atgctggtgt atctagacca aggtttacaa 4046
actgcacggt gatcacgcag ttatcgtgat tatcacgtgc aacaaccttc tcagttttaa 4106
aaccacgtta tatctccacg gtaaccttgt ggttttcatg ataaccgtga taaccatcaa 4166
acggtgaggt ggtggtaacc caccccaaac gatt 4200
<210>2
<211>662
<212>PRT
<213〉Oryza rice strain WS59
<400>2
Met Val Met Gly Cys Thr His Gln Gln Leu Ala Asp Pro Trp Ala Trp
1 5 10 15
Ala Ala Arg Glu Glu Gly Gly Arg Val Ala Gly Ser Ser Leu Pro Arg
20 25 30
Arg Pro Thr Pro Thr Thr Ala Ile Gly Arg Val Ala Ala Gly Arg Arg
35 40 45
Pro Pro Ala Pro Ala Thr Gly Pro Pro Gln Gly Thr Pro Ala Met Ser
50 55 60
Leu Ala Pro Pro Pro Pro Lys Ala Ser Pro Ser Ala Ala Arg Leu Gln
65 70 75 80
Lys Pro Ala Ala Ser Cys His Gly Gly Gln Pro Lys Ala Ser Pro Trp
85 90 95
Ala Gly Ser Pro Pro Gly Ala Ala Thr His Leu Gln Gln Pro Ser Arg
100 105 110
Met Lys Leu Ser Pro Ser Pro Ala Pro Gly Ala Gln His Leu Pro Pro
115 120 125
Ala Lys Pro Ser Pro Leu Ala Gly Ser Pro Pro Gly Ala Ala Thr His
130 135 140
Leu Gln Gln Pro Ala Ala Arg Met Leu Ala Ser Ser Pro Ala Lys Arg
145 150 155 160
Val Leu Ser Val Phe Ser Pro Ser Ala Ala Gly Ser Leu Arg Arg Glu
165 170 175
Arg Asn His Leu Pro Pro Leu Ala Ser Cys Phe Cys Leu Arg Arg Leu
180 185 190
Met Ser Thr Gly Leu Pro Gln Ile Pro Ser Pro Pro Ala Thr Leu Met
195 200 205
Arg Pro Trp Cys Asn Glu Asp Glu Gln Val Pro Gln Leu Thr Pro Phe
210 215 220
Lys Ile Ser His Gly Val Leu Asp Thr Met Met Lys Gln Tyr Glu Glu
225 230 235 240
Asp Tyr Lys Tyr Leu Val Glu Glu Gly Leu Asp Leu Glu Glu Met Lys
245 250 255
Lys Leu Tyr His Glu Leu Val Arg Pro Val Leu Val Ala Ser Cys Leu
260 265 270
Ile Leu Glu His His Ile Val Ser Pro Glu Thr Lys Ala Leu Asp Gln
275 280 285
Lys Phe Lys Asp Lys Phe Gly Lys Pro Pro Ile Tyr Leu Ser Ala Leu
290 295 300
Asp Ile Met Lys Thr Leu Ile Ala Met Glu Pro Ser Arg Trp Ser Tyr
305 310 315 320
Leu Met Asp Phe Leu Arg Glu Lys Arg Ile Leu Val Thr Glu Asn Trp
325 330 335
Glu Val Ser Leu Gly Ser Ile Glu Cys Ile Met His Asp Asp Ser Ile
340 345 350
Ser Phe Met Met Asn Gly Arg Val Val Phe Lys Gly Gln Ser Asp Gly
355 360 365
Arg Tyr Asn His Gly Thr Gly Thr Ala Asp Leu Cys Thr Ala Ile Thr
370 375 380
Gly Ala Ala Thr Asp Ile Val Asp Cys Ala Ile Tyr Arg Glu Val Pro
385 390 395 400
Cys Asn Ser Pro Thr His Ala Glu Leu Leu Gly Met Tyr Val Leu Glu
405 410 415
Arg Arg Ala Ile Ser Leu Lys Ile Leu Leu Phe Asp Val Lys Thr Asp
420 425 430
Asn Ala Phe Val Ser Glu Thr Val Arg Asp Met Phe Pro Ile Thr Pro
435 440 445
Asn Thr Thr Glu Lys Asp Leu Cys Gln Val Leu Arg Ser Met Lys Val
450 455 460
Tyr Phe Glu His Phe Asn Cys Arg Cys Glu Pro Arg Glu Lys Leu Glu
465 470 475 480
Leu Val Asp Ser Leu Met Lys Met Lys Asp Asn Glu Leu Thr Met Glu
485 490 495
Ser Ile Lys Asp Lys Trp Ala His Ser Leu Leu Arg Leu Pro Val Phe
500 505 510
Arg Ala His Gln Pro Thr Lys Thr Ile Arg Lys Asp Tyr Ile Asn Lys
515 520 525
Ala Pro Thr Val Gly Thr Val Ala Phe His Gly Gln Tyr Lys Ile Val
530 535 540
Ala Lys Gly Asp Thr Ala Lys Thr Glu Ala Val Val Asn Leu Val Leu
545 550 555 560
Ser Leu Gln Pro Met Val Ala Ser Val Phe Val Gly Asn His Ile Ser
565 570 575
Ala Ile Ser Val Lys Glu Glu Ile Glu Lys Phe Val Lys Gly Tyr Leu
580 585 590
Leu Asp Ile Lys Glu Ala Lys Gly Ala Tyr Val Val Ile Met Lys Phe
595 600 605
Ile Glu Thr Pro Asn Ala Gly Gly Lys Ile Leu Asp Val Leu Phe Asn
610 615 620
Thr Glu Val Gly Glu Val Tyr Lys Gln Thr Ala Asn Ser Val Cys Val
625 630 635 640
Lys Leu Leu Asp Lys Asn Asp Gly Thr Thr Pro Glu Leu Ser Pro Ala
645 650 655
Cys Leu Leu Ile Phe Gly
660
<210>3
<211>4200
<212>DNA
<213〉Oryza rice strain Dular
<220>
<221>CDS
<222>(1293)..(1862)
<220>
<221>CDS
<222>(1997)..(3005)
<220>
<221>CDS
<222>(3091)..(3500)
<400>3
aaaatcatcc tgttgagggt gttgtagcaa attttatctt tttttcatac atgcaatttc 60
ataaattttt ataaactgta atttttttaa tgtttaggaa ggagtagtct ctttgtgttt 120
tcctttcaag agtaaactgc acagacagtc cctaaacttg tgatggtgta tcacataggt 180
ccctaaactc tcaaaatgta ttttttgtgt tcccgaactt gtctagggtg tcgtataggt 240
ccaaacaagc gctaatccga cccatgtgtt gacttgacat gtcacaatgg tttctacatg 300
ttggtgggac ccacatgtca atcacataaa aaaattgaag gataaaggag acaaaaatgt 360
ttttttttaa ttttgaaaaa aaaagagagg agagagagag atgagaaaaa gatgttttta 420
aatttttatt ctatgtgtgt ttgataagtg ggtcccacta acacgtaggt gccactatga 480
tattgtcatg tcagtgtaca aacctacatg acagtcaaga taagtttgga gacaagaaga 540
aaagaatttt aagagtttag ggatatagat gatatactac gcatgtgcac ttcactcgtt 600
ttgcaatgtt gcaattgatt ttccttttta tgtttcacgt gctggggtct tggtggtgtt 660
gcaacaagta ttgttgactg ttgcatctgt tagaaatatg catatgcatg tttcatcgag 720
tgaggaacta tttgaaacag ctagcggggt ttgaaatttc gtttcaaaat ttccgaaatt 780
tcagtaatct catatatggc ccgaaatttt cggttttttg caaatcttta tgaatttggt 840
caaattttat tcaaattcat tcaaaatcag tcaaaatgtt aaataatttc ggccaaaaaa 900
aaaaagaaat tcccgaaatt tctgtgcccc cggcagaaat agcaatttcg aaatttaaaa 960
ctgtgagcta gccaaggaaa ttaattaagc acaaacagcg cagcttagcg aagtgagtga 1020
gtgaaggagt gcgcgcatcg cttcgcctgg agagaccacc ttcttctcca atccccagtt 1080
ttctcccctt gctgcttttg cccgcccccc tctccccccc aaaaaaaacc aaaaaaaaaa 1140
aaagaagaag aagaagaaga agaagaaagg aagaagaaaa cctagctagt gtgaatcgcg 1200
agcacttacc gatgcttcga agggcgacgg ctgcggtggc cggccgccgt ctacgaccgg 1260
cggtgggcct ctcgccgtcg gccccggcga ga atg gtg atg ggt tgc acc cac 1313
Met Val Met Gly Cys Thr His
1 5
cag cag ctc gcc gac cca tgg gca tgg gca gcc cgc gag gaa ggt ggc 1361
Gln Gln Leu Ala Asp Pro Trp Ala Trp Ala Ala Arg Glu Glu Gly Gly
10 15 20
cgt gtc gcc ggc agc tcc ctg cca cgg cgg ccg acc cct acc acc gcc 1409
Arg Val Ala Gly Ser Ser Leu Pro Arg Arg Pro Thr Pro Thr Thr Ala
25 30 35
atc ggc cgg gtc gcc gcc ggg cgc cgc cca cct gca ccg gca act ggc 1457
Ile Gly Arg Val Ala Ala Gly Arg Arg Pro Pro Ala Pro Ala Thr Gly
40 45 50 55
cca ccg cag ggg acg ccc gcc atg tcg ctg gca cca cca cca cca aag 1505
Pro Pro Gln Gly Thr Pro Ala Met Ser Leu Ala Pro Pro Pro Pro Lys
60 65 70
gcc tcg cct tcc gcc gcc cgc ctg cag aag ccg gca gct tcc tgc cac 1553
Ala Ser Pro Ser Ala Ala Arg Leu Gln Lys Pro Ala Ala Ser Cys His
75 80 85
ggc ggc caa cca aag gcc tcg cct tgg gcc ggg tcg cca ccg gga gca 1601
Gly Gly Gln Pro Lys Ala Ser Pro Trp Ala Gly Ser Pro Pro Gly Ala
90 95 100
gcc acc cac ctg cag cag ccg tcg cgg atg aag ctc tct ccg tcg ccg 1649
Ala Thr His Leu Gln Gln Pro Ser Arg Met Lys Leu Ser Pro Ser Pro
105 110 115
gca cct ggc gcc caa cac ctg cca ccg gca aag ccc tcg cct ttg gcc 1697
Ala Pro Gly Ala Gln His Leu Pro Pro Ala Lys Pro Ser Pro Leu Ala
120 125 130 135
ggg tcg ccg ccg gga gca gcc acc cac ctg cag cag ccg gcg gcg agg 1745
Gly Ser Pro Pro Gly Ala Ala Thr His Leu Gln Gln Pro Ala Ala Arg
140 145 150
atg ctg gcc tcg tcg ccg gcg aag agg gtg ctc tca gtc ttc tca cca 1793
Met Leu Ala Ser Ser Pro Ala Lys Arg Val Leu Ser Val Phe Ser Pro
155 160 165
tca gcg gcc ggg tcg ctg cgg agg gaa cgt aac cat ctg ccg cca ttg 1841
Ser Ala Ala Gly Ser Leu Arg Arg Glu Arg Asn His Leu Pro Pro Leu
170 175 180
gcc agc tgc ttc tgc ttg cgg gtgggtaact ccaaattcca aatttaggtc 1892
Ala Ser Cys Phe Cys Leu Arg
185 190
caattaaagg acccccaatt tcctcgcttg tagctagtct tatcttattt gggtcatgta 1952
ttcattgact gatgatttgg gtcgtgtata tattgtgttt gcag cgg ctg atg agt 2008
Arg Leu Met Ser
acc gga cta cct caa att cca agt ccc cca gca aca ctg atg aga cct 2056
Thr Gly Leu Pro Gln Ile Pro Ser Pro Pro Ala Thr Leu Met Arg Pro
195 200 205 210
tgg tgt aat gag gat gag cag gtg cct caa cta acc cct ttt aag atc 2104
Trp Cys Asn Glu Asp Glu Gln Val Pro Gln Leu Thr Pro Phe Lys Ile
215 220 225
agt cat ggt gtc tcg gat aca atg atg aag cag tat gag gaa gac tac 2152
Ser His Gly Val Ser Asp Thr Met Met Lys Gln Tyr Glu Glu Asp Tyr
230 235 240
aag tat ctt gtg gag gag ggg ctg gat ctt gaa gag atg aag aaa ctt 2200
Lys Tyr Leu Val Glu Glu Gly Leu Asp Leu Glu Glu Met Lys Lys Leu
245 250 255
tac cat gag ctg gtg cgt ccc gtt tta gtt gca agc tgt ctg att ctt 2248
Tyr His Glu Leu Val Arg Pro Val Leu Val Ala Ser Cys Leu Ile Leu
260 265 270
gaa cat cac att gtc agt cct gaa acc aaa gcg cta gat cag aaa ttc 2296
Glu His His Ile Val Ser Pro Glu Thr Lys Ala Leu Asp Gln Lys Phe
275 280 285 290
aaa gac aag ttc ggg aaa ccc cca atc tat ctc tca gct ctt gat ata 2344
Lys Asp Lys Phe Gly Lys Pro Pro Ile Tyr Leu Ser Ala Leu Asp Ile
295 300 305
atg aag acc ttg att gct atg gag ccg tcg aga tgg agt tac ctt atg 2392
Met Lys Thr Leu Ile Ala Met Glu Pro Ser Arg Trp Ser Tyr Leu Met
310 315 320
gat ttc tta aga gag aaa cgt ata ttg gtc act gag aat tgg gaa gtt 2440
Asp Phe Leu Arg Glu Lys Arg Ile Leu Val Thr Glu Asn Trp Glu Val
325 330 335
tca ctg ggt tcc atc gag tgc atc atg cac gac gac agc att agt ttt 2488
Ser Leu Gly Ser Ile Glu Cys Ile Met His Asp Asp Ser Ile Ser Phe
340 345 350
atg atg aat ggc cgg gtc gtt ttc aaa ggg caa tct gat ggt agg tat 2536
Met Met Asn Gly Arg Val Val Phe Lys Gly Gln Ser Asp Gly Arg Tyr
355 360 365 370
aac cat ggg act ggt aca gcc gat ctt tgc aca acg att act ggt gta 2584
Asn His Gly Thr Gly Thr Ala Asp Leu Cys Thr Thr Ile Thr Gly Val
375 380 385
gct aca gat atc gtt gat tgt gcc atc tac cga gag gtc cct tgc aat 2632
Ala Thr Asp Ile Val Asp Cys Ala Ile Tyr Arg Glu Val Pro Cys Asn
390 395 400
tct cca acg cac gct gag ttg ctt gga atg tat gtt cta gaa aga agg 2680
Ser Pro Thr His Ala Glu Leu Leu Gly Met Tyr Val Leu Glu Arg Arg
405 410 415
gca ata tcg ctg aaa ata ctg ttg ttt gat gtg aaa acc gac aac gct 2728
Ala Ile Ser Leu Lys Ile Leu Leu Phe Asp Val Lys Thr Asp Asn Ala
420 425 430
ttt gtc agt gaa aca gtc aga gat atg ttt cct att act cca aac act 2776
Phe Val Ser Glu Thr Val Arg Asp Met Phe Pro Ile Thr Pro Asn Thr
435 440 445 450
act gag aag gat ctg tgc caa gtg ttg aga agt atg aaa gtc tat ttc 2824
Thr Glu Lys Asp Leu Cys Gln Val Leu Arg Ser Met Lys Val Tyr Phe
455 460 465
gaa cac ttc aat tgt cgg tgt gaa cct cgg gag aaa ctt gag cta gtg 2872
Glu His Phe Asn Cys Arg Cys Glu Pro Arg Glu Lys Leu Glu Leu Val
470 475 480
gat agt tta atg aaa atg aag gat aat gag ttg aca atg gaa tca atc 2920
Asp Ser Leu Met Lys Met Lys Asp Asn Glu Leu Thr Met Glu Ser Ile
485 490 495
aag gat aaa tgg gct cat tac ctt ctg aga ttg cct gtc ttc agg gct 2968
Lys Asp Lys Trp Ala His Tyr Leu Leu Arg Leu Pro Val Phe Arg Ala
500 505 510
cat cag cca acc aag acc ata aga aag gac tat ata a gtaagtagtt 3015
His Gln Pro Thr Lys Thr Ile Arg Lys Asp Tyr Ile
515 520 525
aatcttctga gtacattgac tttgattttt ttttcttttt tcttttaacc catcggttta 3075
atttttatat tgcag ac aag gct cca act gtt ggt aca gta gct ttc cat 3125
Asn Lys Ala Pro Thr Val Gly Thr Val Ala Phe His
530 535
ggg caa tac aaa att gtc gca aag agg gat aca gca aaa acg gag gct 3173
Gly Gln Tyr Lys Ile Val Ala Lys Arg Asp Thr Ala Lys Thr Glu Ala
540 545 550
gtt gtg aac ttg gtg cta tcg ctt cag cca atg gtt gct tca gtt ttc 3221
Val Val Asn Leu Val Leu Ser Leu Gln Pro Met Val Ala Ser Val Phe
555 560 565 570
gtg gga aat cac atc tca gca atc tca gtc aag gaa gag att gaa aag 3269
Val Gly Asn His Ile Ser Ala Ile Ser Val Lys Glu Glu Ile Glu Lys
575 580 585
ttt gtc aag ggg tat tta ctg gac atc aag gag gct aag gga gca tat 3317
Phe Val Lys Gly Tyr Leu Leu Asp Ile Lys Glu Ala Lys Gly Ala Tyr
590 595 600
gtt gtt atc atg aaa ttt ata gaa act ccc aat gca ggt ggt aaa ata 3365
Val Val Ile Met Lys Phe Ile Glu Thr Pro Asn Ala Gly Gly Lys Ile
605 610 615
tta gat gtc ttg ttt aat act gaa gtc ggt gag gtg tac aag cag aca 3413
Leu Asp Val Leu Phe Asn Thr Glu Val Gly Glu Val Tyr Lys Gln Thr
620 625 630
gca aac tcg gtc tgc gtc aaa cta ctg gac aag aat gat gga act act 3461
Ala Asn Ser Val Cys Val Lys Leu Leu Asp Lys Asn Asp Gly Thr Thr
635 640 645 650
cca gag cta agt cca gca tgt ctt cgt att ttc ggt tgagtcacaa 3507
Pro Glu Leu Ser Pro Ala Cys Leu Arg Ile Phe Gly
655 660
agggtcaaac tagtagtttc ttggcatgtg ccttctgtaa gtgtgctgtt taggtggctg 3567
gagggccctt tttggttagt agtctagacc ctaacaacat gttattactt atgctgctgt 3627
tatgcaccct gtgcagcacg cttctgaatt gcaacttctt ttcaagctga atttatctgc 3687
gagtgttaac cctattttgt gatggtcata tatgtttatg gatggagctt caagctgggg 3747
catctgcagg gatctgagta ccagaactga agttctttgt gaaagaagtt gttttgcttg 3807
tagatggcaa tttccatgca gtctcatctg tacaaccccc aaaaagcatc attctgtttc 3867
agctacgagc tctgtgtttg tgcttttgtt tttcttcttg tttaccccag gatgggatgg 3927
caatggtagg gaatgctggt gtatctagac cgtgtttgtg cttttgtttt tcttcttgtt 3987
taccccagga tgggacggca atggtaggga atgctggtgt atctagacca aggtttacaa 4047
actgcacggt gatcacgcag ttatcgtgat tatcacgtgc aacaaccttc ttaaccttgt 4107
ggttttcatg ataaccgtga taaccatcaa acggtgaggt ggtggtaacc caccccaaac 4167
gattcggtga accttgatct agacaggtga tct 4200
<210>4
<211>662
<212>PRT
<213〉Oryza rice strain Dular
<400>4
Met Val Met Gly Cys Thr His Gln Gln Leu Ala Asp Pro Trp Ala Trp
1 5 10 15
Ala Ala Arg Glu Glu Gly Gly Arg Val Ala Gly Ser Ser Leu Pro Arg
20 25 30
Arg Pro Thr Pro Thr Thr Ala Ile Gly Arg Val Ala Ala Gly Arg Arg
35 40 45
Pro Pro Ala Pro Ala Thr Gly Pro Pro Gln Gly Thr Pro Ala Met Ser
50 55 60
Leu Ala Pro Pro Pro Pro Lys Ala Ser Pro Ser Ala Ala Arg Leu Gln
65 70 75 80
Lys Pro Ala Ala Ser Cys His Gly Gly Gln Pro Lys Ala Ser Pro Trp
85 90 95
Ala Gly Ser Pro Pro Gly Ala Ala Thr His Leu Gln Gln Pro Ser Arg
100 105 110
Met Lys Leu Ser Pro Ser Pro Ala Pro Gly Ala Gln His Leu Pro Pro
115 120 125
Ala Lys Pro Ser Pro Leu Ala Gly Ser Pro Pro Gly Ala Ala Thr His
130 135 140
Leu Gln Gln Pro Ala Ala Arg Met Leu Ala Ser Ser Pro Ala Lys Arg
145 150 155 160
Val Leu Ser Val Phe Ser Pro Ser Ala Ala Gly Ser Leu Arg Arg Glu
165 170 175
Arg Asn His Leu Pro Pro Leu Ala Ser Cys Phe Cys Leu Arg Arg Leu
180 185 190
Met Ser Thr Gly Leu Pro Gln Ile Pro Ser Pro Pro Ala Thr Leu Met
195 200 205
Arg Pro Trp Cys Asn Glu Asp Glu Gln Val Pro Gln Leu Thr Pro Phe
210 215 220
Lys Ile Ser His Gly Val Ser Asp Thr Met Met Lys Gln Tyr Glu Glu
225 230 235 240
Asp Tyr Lys Tyr Leu Val Glu Glu Gly Leu Asp Leu Glu Glu Met Lys
245 250 255
Lys Leu Tyr His Glu Leu Val Arg Pro Val Leu Val Ala Ser Cys Leu
260 265 270
Ile Leu Glu His His Ile Val Ser Pro Glu Thr Lys Ala Leu Asp Gln
275 280 285
Lys Phe Lys Asp Lys Phe Gly Lys Pro Pro Ile Tyr Leu Ser Ala Leu
290 295 300
Asp Ile Met Lys Thr Leu Ile Ala Met Glu Pro Ser Arg Trp Ser Tyr
305 310 315 320
Leu Met Asp Phe Leu Arg Glu Lys Arg Ile Leu Val Thr Glu Asn Trp
325 330 335
Glu Val Ser Leu Gly Ser Ile Glu Cys Ile Met His Asp Asp Ser Ile
340 345 350
Ser Phe Met Met Asn Gly Arg Val Val Phe Lys Gly Gln Ser Asp Gly
355 360 365
Arg Tyr Asn His Gly Thr Gly Thr Ala Asp Leu Cys Thr Thr Ile Thr
370 375 380
Gly Val Ala Thr Asp Ile Val Asp Cys Ala Ile Tyr Arg Glu Val Pro
385 390 395 400
Cys Asn Ser Pro Thr His Ala Glu Leu Leu Gly Met Tyr Val Leu Glu
405 410 415
Arg Arg Ala Ile Ser Leu LysIle Leu Leu Phe Asp Val Lys Thr Asp
420 425 430
Asn Ala Phe Val Ser Glu Thr Val Arg Asp Met Phe Pro Ile Thr Pro
435 440 445
Asn Thr Thr Glu Lys Asp Leu Cys Gln Val Leu Arg Ser Met Lys Val
450 455 460
Tyr Phe Glu His Phe Asn Cys Arg Cys Glu Pro Arg Glu Lys Leu Glu
465 470 475 480
Leu Val Asp Ser Leu Met Lys Met Lys Asp Asn Glu Leu Thr Met Glu
485 490 495
Ser Ile Lys Asp Lys Trp Ala His Tyr Leu Leu Arg Leu Pro Val Phe
500 505 510
Arg Ala His Gln Pro Thr Lys Thr Ile Arg Lys Asp Tyr Ile Asn Lys
515 520 525
Ala Pro Thr Val Gly Thr Val Ala Phe His Gly Gln Tyr Lys Ile Val
530 535 540
Ala Lys Arg Asp Thr Ala Lys Thr Glu Ala Val Val Asn Leu Val Leu
545 550 555 560
Ser Leu Gln Pro Met Val Ala Ser Val Phe Val Gly Asn His Ile Ser
565 570 575
Ala Ile Ser Val Lys Glu Glu Ile Glu Lys Phe Val Lys Gly Tyr Leu
580 585 590
Leu Asp Ile Lys Glu Ala Lys Gly Ala Tyr Val Val Ile Met Lys Phe
595 600 605
Ile Glu Thr Pro Asn Ala Gly Gly Lys Ile Leu Asp Val Leu Phe Asn
610 615 620
Thr Glu Val Gly Glu Val Tyr Lys Gln Thr Ala Asn Ser Val Cys Val
625 630 635 640
Lys Leu Leu Asp Lys Asn Asp Gly Thr Thr Pro Glu Leu Ser Pro Ala
645 650 655
Cys Leu Arg Ile Phe Gly
660
<210>5
<211>4200
<212>DNA
<213〉the Oryza rice strain 02428
<220>
<221>CDS
<222>(1293)..(1862)
<220>
<221>CDS
<222>(1997)..(3005)
<220>
<221>CDS
<222>(3091)..(3500)
<400>5
aaaatcatcc tgttgagggt gttgtagcaa attttatctt tttttcatac atgcaatttc 60
ataaattttt ataaactgta atttttttaa tgtttaggaa ggagtagtct ctttgtgttt 120
tcctttcaag agtaaactgc acagacagtc cctaaacttg tgatggtgta tcacataggt 180
ccctaaactc tcaaaatgta ttttttgtgt tcccgaactt gtctagggtg tcgtataggt 240
ccaaacaagc gctaatccga cccatgtgtt gacttgacat gtcacaatgg tttctacatg 300
ttggtgggac ccacatgtca atcacataaa aaaattgaag gataaaggag acaaaaatgt 360
ttttttttaa ttttgaaaaa aaaagagagg agagagagag atgagaaaaa gatgttttta 420
aatttttatt ctatgtgtgt ttgataagtg ggtcccacta acacgtaggt gccactatga 480
tattgtcatg tcagtgtaca aacctacatg acagtcaaga taagtttgga gacaagaaga 540
aaagaatttt aagagtttag ggatatagat gatatactac gcatgtgcac ttcactcgtt 600
ttgcaatgtt gcaattgatt ttccttttta tgtttcacgt gctggggtct tggtggtgtt 660
gcaacaagta ttgttgactg ttgcatctgt tagaaatatg catatgcatg tttcatcgag 720
tgaggaacta tttgaaacag ctagcggggt ttgaaatttc gtttcaaaat ttccgaaatt 780
tcagtaatct catatatggc ccgaaatttt cggttttttg caaatcttta tgaatttggt 840
caaattttat tcaaattcat tcaaaatcag tcaaaatgtt aaataatttc ggccaaaaaa 900
aaaaagaaat tcccgaaatt tctgtgcccc cggcagaaat agcaatttcg aaatttaaaa 960
ctgtgagcta gccaaggaaa ttaattaagc acaaacagcg cagcttagcg aagtgagtga 1020
gtgaaggagt gcgcgcatcg cttcgcctgg agagaccacc ttcttctcca atccccagtt 1080
ttctcccctt gctgcttttg cccgcccccc tctccccccc aaaaaaaacc aaaaaaaaaa 1140
aaagaagaag aagaagaaga agaagaaagg aagaagaaaa cctagctagt gtgaatcgcg 1200
agcacttacc gatgcttcga agggcgacgg ctgcggtggc cggccgccgt ctacgaccgg 1260
cggtgggcct ctcgccgtcg gccccggcga ga atg gtg atg ggt tgc acc cac 1313
Met Val Met Gly Cys Thr His
1 5
cag cag ctc gcc gac cca tgg gca tgg gca gcc cgc gag gaa ggt ggc 1361
Gln Gln Leu Ala Asp Pro Trp Ala Trp Ala Ala Arg Glu Glu Gly Gly
10 15 20
cgt gtc gcc ggc agc tcc ctg cca cgg cgg ccg acc cct acc acc gcc 1409
Arg Val Ala Gly Ser Ser Leu Pro Arg Arg Pro Thr Pro Thr Thr Ala
25 30 35
atc ggc cgg gtc gcc gcc ggg cgc cgc cca cct gca ccg gca act ggc 1457
Ile Gly Arg Val Ala Ala Gly Arg Arg Pro Pro Ala Pro Ala Thr Gly
40 45 50 55
cca ccg cag ggg acg ccc gcc atg tcg ctg gca cca cca cca cca aag 1505
Pro Pro Gln Gly Thr Pro Ala Met Ser Leu Ala Pro Pro Pro Pro Lys
60 65 70
gcc tcg cct tcc gcc gcc cgc ctg cag aag ccg gca gct tcc tgc cac 1553
Ala Ser Pro Ser Ala Ala Arg Leu Gln Lys Pro Ala Ala Ser Cys His
75 80 85
ggc ggc caa cca aag gcc tcg cct tgg gcc ggg tcg ccg ccg gga gca 1601
Gly Gly Gln Pro Lys Ala Ser Pro Trp Ala Gly Ser Pro Pro Gly Ala
90 95 100
gcc acc cac ctg cag cag ccg tcg cgg atg aag ctc tct ccg tcg ccg 1649
Ala Thr His Leu Gln Gln Pro Ser Arg Met Lys Leu Ser Pro Ser Pro
105 110 115
gca cct ggc gcc caa cac ctg cca ccg gca aag ccc tcg cct ttg gcc 1697
Ala Pro Gly Ala Gln His Leu Pro Pro Ala Lys Pro Ser Pro Leu Ala
120 125 130 135
ggg tcg ccg ccg gga gca gcc acc cac ctg cag cag ccg gcg gcg agg 1745
Gly Ser Pro Pro Gly Ala Ala Thr His Leu Gln Gln Pro Ala Ala Arg
140 145 150
atg ctg gcc tcg tcg ccg gcg aag agg gtg ctc tca gtc ttc tca cca 1793
Met Leu Ala Ser Ser Pro Ala Lys Arg Val Leu Ser Val Phe Ser Pro
155 160 165
tca gcg gcc ggg tcg ctg cgg agg gaa cgt aac cat cta ccg cca ttg 1841
Ser Ala Ala Gly Ser Leu Arg Arg Glu Arg Asn His Leu Pro Pro Leu
170 175 180
gcc agc tgc ttc tgc ttg cgg gtgggtaact ccaaattcca aatttaggtc 1892
Ala Ser Cys Phe Cys Leu Arg
185 190
caattaaagg acccccaatt tcctcgcttg tagctagtct tatcttattt gggtcatgta 1952
ttcattgact gatgatttgg gtcgtgtata tattgtgttt gcag cgg ctg atg agt 2008
Arg Leu Met Ser
acc gga cta cct caa att cca agt ccc cca gca aca ctg atg aga cct 2056
Thr Gly Leu Pro Gln Ile Pro Ser Pro Pro Ala Thr Leu Met Arg Pro
195 200 205 210
tgg tgt aat gag gat gag cag gtg cct caa cta acc cct ttt aag atc 2104
Trp Cys Asn Glu Asp Glu Gln Val Pro Gln Leu Thr Pro Phe Lys Ile
215 220 225
agt cat ggt gtc tcg gat aca atg atg aag cag tat gag gaa gac tac 2152
Ser His Gly Val Ser Asp Thr Met Met Lys Gln Tyr Glu Glu Asp Tyr
230 235 240
aag tat ctt gtg gag gag ggg ctg gat ctt gaa gag atg aag aaa ctt 2200
Lys Tyr Leu Val Glu Glu Gly Leu Asp Leu Glu Glu Met Lys Lys Leu
245 250 255
tac cat gag ctg gtg cgt ccc gtt tta gtt gca agc tgt ctg att ctt 2248
Tyr His Glu Leu Val Arg Pro Val Leu Val Ala Ser Cys Leu Ile Leu
260 265 270
gaa cat cac att gtc agt cct gaa acc aaa gcg cta gat cag aaa tcc 2296
Glu His His Ile Val Ser Pro Glu Thr Lys Ala Leu Asp Gln Lys Ser
275 280 285 290
aaa gac aag ttc ggg aaa ccc cca atc tat ctc tca gct ctt gat ata 2344
Lys Asp Lys Phe Gly Lys Pro Pro Ile Tyr Leu Ser Ala Leu Asp Ile
295 300 305
atg aag acc ttg att gct atg gag ccg tcg aga tgg agt tac ctt atg 2392
Met Lys Thr Leu Ile Ala Met Glu Pro Ser Arg Trp Ser Tyr Leu Met
310 315 320
gat ttc tta aga gag aaa cgt ata ttg gtc act gag aat tgg gaa gtt 2440
Asp Phe Leu Arg Glu Lys Arg Ile Leu Val Thr Glu Asn Trp Glu Val
325 330 335
tca ctg ggt tcc atc gag tgc atc atg cac gac gac agc att agt ttt 2488
Ser Leu Gly Ser Ile Glu Cys Ile Met His Asp Asp Ser Ile Ser Phe
340 345 350
atg atg aat ggc cgg gtc gtt ttc aaa ggg caa tct gat ggt agg tat 2536
Met Met Asn Gly Arg Val Val Phe Lys Gly Gln Ser Asp Gly Arg Tyr
355 360 365 370
aac cat ggg act ggt aca gcc gat ctt tgc aca gcg att act ggt gaa 2584
Asn His Gly Thr Gly Thr Ala Asp Leu Cys Thr Ala Ile Thr Gly Glu
375 380 385
gct aca gat atc gtt gat tgt gcc atc tac cga gag gtc cct tgc aat 2632
Ala Thr Asp Ile Val Asp Cys Ala Ile Tyr Arg Glu Val Pro Cys Asn
390 395 400
tct cca acg cac gct gag ttg ctt gga atg tat gtt cta gaa aga agg 2680
Ser Pro Thr His Ala Glu Leu Leu Gly Met Tyr Val Leu Glu Arg Arg
405 410 415
gca ata tcg ctg aaa ata ctg ttg ttt gat gtg aaa acc gac aac gct 2728
Ala Ile Ser Leu Lys Ile Leu Leu Phe Asp Val Lys Thr Asp Asn Ala
420 425 430
ttt gtc agt gaa aca gtc aga gat atg ttt cct att act cca aac act 2776
Phe Val Ser Glu Thr Val Arg Asp Met Phe Pro Ile Thr Pro Asn Thr
435 440 445 450
act gag aag gat ctg tgc caa gtg ttg aga agt atg aaa gtc tat ttc 2824
Thr Glu Lys Asp Leu Cys Gln Val Leu Arg Ser Met Lys Val Tyr Phe
455 460 465
gaa cac ttc aat tgt cgg tgt gaa cct cgg gag aaa ctt gag cta gtg 2872
Glu His Phe Asn Cys Arg Cys Glu Pro Arg Glu Lys Leu Glu Leu Val
470 475 480
gat agt tta atg aaa atg aag gat aat gag atg aca atg gaa tca atc 2920
Asp Ser Leu Met Lys Met Lys Asp Asn Glu Met Thr Met Glu Ser Ile
485 490 495
aag gat aaa tgg gct cat tac ctt ctg aga ttg cct gtc ttc agg gct 2968
Lys Asp Lys Trp Ala His Tyr Leu Leu Arg Leu Pro Val Phe Arg Ala
500 505 510
cat cag cca acc aag acc ata aga aag gac tat ata a gtaagtagtt 3015
His Gln Pro Thr Lys Thr Ile Arg Lys Asp Tyr Ile
515 520 525
aatcttctga gtacattgac tttgattttt ttttcttttt tcttttaacc catcggttta 3075
atttttatat tgcag ac aag gct cca act gtt ggt aca gta gct ttc cat 3125
Asn Lys Ala Pro Thr Val Gly Thr Val Ala Phe His
530 535
ggg caa tac aaa att gtc gca aag ggg gat aca gca aaa acg gag gct 3173
Gly Gln Tyr Lys Ile Val Ala Lys Gly Asp Thr Ala Lys Thr Glu Ala
540 545 550
gtt gtg aac ttg gtg cta tcg ctt cag cca atg gtt gct tca gtt ttc 3221
Val Val Asn Leu Val Leu Ser Leu Gln Pro Met Val Ala Ser Val Phe
555 560 565 570
gtg gga aat cac atc tca gca atc tca gtc aag gaa gag ata gaa aag 3269
Val Gly Asn His Ile Ser Ala Ile Ser Val Lys Glu Glu Ile Glu Lys
575 580 585
ttt gtc aag ggg tat tta ctg gac atc aag gag gct aag gga gca tat 3317
Phe Val Lys Gly Tyr Leu Leu Asp Ile Lys Glu Ala Lys Gly Ala Tyr
590 595 600
gtt gtt gtc atg aaa ttt ata gaa act ccc aat gca ggt ggt aaa ata 3365
Val Val Val Met Lys Phe Ile Glu Thr Pro Asn Ala Gly Gly Lys Ile
605 610 615
tta gat gtc ttg ttt aat gct gaa gtc ggt gag gtg tac aag cag aca 3413
Leu Asp Val Leu Phe Asn Ala Glu Val Gly Glu Val Tyr Lys Gln Thr
620 625 630
gca aac tcg gtc tgc gtc aaa cta ctg gac aag aat gat gga act act 3461
Ala Asn Ser Val Cys Val Lys Leu Leu Asp Lys Asn Asp Gly Thr Thr
635 640 645 650
cca gag cta agt cca gca tgt ctt ctt att ttc ggt taagtcacaa 3507
Pro Glu Leu Ser Pro Ala Cys Leu Leu Ile Phe Gly
655 660
agggtcaaac tagtagtttc ttggcatgtg ccttctgtaa gtgtgctgtt taggtggctg 3567
gagggccctt tttggttagt agtctagacc ctaacaacat gttattactt atgctgctgt 3627
tatgcaccct gtgcagcacg cttctgaatt gcaacttctt ttcaagctga atttatctgc 3687
gagtgttaac cctattttgt gatggtcata tatgtttatg gatggagctt caagctgggg 3747
catctgcagg gatctgagta ccagaactga agttctttgt gaaagaagtt gttttgcttg 3807
tagatggcaa tttccatgca gtctcatctg tacaaccccc aaaaagcatc attctgtttc 3867
agctacgagc tctgtgtttg tgcttttgtt tttcttcttg tttaccccag gatgggatgg 3927
caatggtagg gaatgctggt gtatctagac cgtgtttgtg cttttgtttt tcttcttgtt 3987
taccccagga tgggacggca atggtaggga atgctggtgt atctagacca aggtttacaa 4047
actgcaeggt gatcacgcag ttatcgtgat tatcacgtgc aacaaccttc ttaaccttgt 4107
ggttttcatg ataacegtga taaccatcaa acggtgaggt ggtggtaacc caccccaaac 4167
gattcggtga accttgatct agacaggtga tct 4200
<210>6
<211>662
<212>PRT
<213〉the Oryza rice strain 02428
<400>6
Met Val Met Gly Cys Thr His Gln Gln Leu Ala Asp Pro Trp Ala Trp
1 5 10 15
Ala Ala Arg Glu Glu Gly Gly Arg Val Ala Gly Ser Ser Leu Pro Arg
20 25 30
Arg Pro Thr Pro Thr Thr Ala Ile Gly Arg Val Ala Ala Gly Arg Arg
35 40 45
Pro Pro Ala Pro Ala Thr Gly Pro Pro Gln Gly Thr Pro Ala Met Ser
50 55 60
Leu Ala Pro Pro Pro Pro Lys Ala Ser Pro Ser Ala Ala Arg Leu Gln
65 70 75 80
Lys Pro Ala Ala Ser Cys His Gly Gly Gln Pro Lys Ala Ser Pro Trp
85 90 95
Ala Gly Ser Pro Pro Gly Ala Ala Thr His Leu Gln Gln Pro Ser Arg
100 105 110
Met Lys Leu Ser Pro Ser Pro Ala Pro Gly Ala Gln His Leu Pro Pro
115 120 125
Ala Lys Pro Ser Pro Leu Ala Gly Ser Pro Pro Gly Ala Ala Thr His
130 135 140
Leu Gln Gln Pro Ala Ala Arg Met Leu Ala Ser Ser Pro Ala Lys Arg
145 150 155 160
Val Leu Ser Val Phe Ser Pro Ser Ala Ala Gly Ser Leu Arg Arg Glu
165 170 175
Arg Asn His Leu Pro Pro Leu Ala Ser Cys Phe Cys Leu Arg Arg Leu
180 185 190
Met Ser Thr Gly Leu Pro Gln Ile Pro Ser Pro Pro Ala Thr Leu Met
195 200 205
Arg Pro Trp Cys Asn Glu Asp Glu Gln Val Pro Gln Leu Thr Pro Phe
210 215 220
Lys Ile Ser His Gly Val Ser Asp Thr Met Met Lys Gln Tyr Glu Glu
225 230 235 240
Asp Tyr Lys Tyr Leu Val Glu Glu Gly Leu Asp Leu Glu Glu Met Lys
245 250 255
Lys Leu Tyr His Glu Leu Val Arg Pro Val Leu Val Ala Ser Cys Leu
260 265 270
Ile Leu Glu His His Ile Val Ser Pro Glu Thr Lys Ala Leu Asp Gln
275 280 285
Lys Ser Lys Asp Lys Phe Gly Lys Pro Pro Ile Tyr Leu Ser Ala Leu
290 295 300
Asp Ile Met Lys Thr Leu Ile Ala Met Glu Pro Ser Arg Trp Ser Tyr
305 310 315 320
Leu Met Asp Phe Leu Arg Glu Lys Arg Ile Leu Val Thr Glu Asn Trp
325 330 335
Glu Val Ser Leu Gly Ser Ile Glu Cys Ile Met His Asp Asp Ser Ile
340 345 350
Ser Phe Met Met Asn Gly Arg Val Val Phe Lys Gly Gln Ser Asp Gly
355 360 365
Arg Tyr Asn His Gly Thr Gly Thr Ala Asp Leu Cys Thr Ala Ile Thr
370 375 380
Gly Glu Ala Thr Asp Ile Val Asp Cys Ala Ile Tyr Arg Glu Val Pro
385 390 395 400
Cys Asn Ser Pro Thr His Ala Glu Leu Leu Gly Met Tyr Val Leu Glu
405 410 415
Arg Arg Ala Ile Ser Leu Lys Ile Leu Leu Phe Asp Val Lys Thr Asp
420 425 430
Asn Ala Phe Val Ser Glu Thr Val Arg Asp Met Phe Pro Ile Thr Pro
435 440 445
Asn Thr Thr Glu Lys Asp Leu Cys Gln Val Leu Arg Ser Met Lys Val
450 455 460
Tyr Phe Glu His Phe Asn Cys Arg Cys Glu Pro Arg Glu Lys Leu Glu
465 470 475 480
Leu Val Asp Ser Leu Met Lys Met Lys Asp Asn Glu Met Thr Met Glu
485 490 495
Ser Ile Lys Asp Lys Trp Ala His Tyr Leu Leu Arg Leu Pro Val Phe
500 505 510
Arg Ala His Gln Pro Thr Lys Thr Ile Arg Lys Asp Tyr Ile Asn Lys
515 520 525
Ala Pro Thr Val Gly Thr Val Ala Phe His Gly Gln Tyr Lys Ile Val
530 535 540
Ala Lys Gly Asp Thr Ala Lys Thr Glu Ala Val Val Asn Leu Val Leu
545 550 555 560
Ser Leu Gln Pro Met Val Ala Ser Val Phe Val Gly Asn His Ile Ser
565 570 575
Ala Ile Ser Val Lys Glu Glu Ile Glu Lys Phe Val Lys Gly Tyr Leu
580 585 590
Leu Asp Ile Lys Glu Ala Lys Gly Ala Tyr Val Val Val Met Lys Phe
595 600 605
Ile Glu Thr Pro Asn Ala Gly Gly Lys Ile Leu Asp Val Leu Phe Asn
610 615 620
Ala Glu Val Gly Glu Val Tyr Lys Gln Thr Ala Asn Ser Val Cys Val
625 630 635 640
Lys Leu Leu Asp Lys Asn Asp Gly Thr Thr Pro Glu Leu Ser Pro Ala
645 650 655
Cys Leu Leu Ile Phe Gly
660

Claims (7)

1, a kind of paddy rice hybrid fertility gene, the sequence of its proteins encoded is:
A) SEQ ID NO:2 or SEQ ID NO:4 or SEQ ID NO:6; Or
B) replace, lack or add the aminoacid sequence with same function of one or several amino-acid residue formation in them.
2, gene according to claim 1, its nucleotides sequence are classified SEQ ID NO:1 or SEQ ID NO:3 or SEQ ID NO:5 as.
3, a kind of carrier that contains claim 1 or 2 described genes.
4, a kind of by the described carrier transformed host cells of claim 3.
5. a method of cultivating the affine system of paddy rice hybrid fertility is characterized in that: with claim 1 or 2 described genes or the described carrier rice transformation of claim 3, obtain the affine system of transgenic paddy rice hybrid fertile; Or select to cultivate the affine system of paddy rice hybrid fertility of containing claim 1 or 2 described genes with hybridizing-backcross method and molecule marker.
6. molecule marker or its close linkage mark that produces according to the nucleotide sequence of claim 1 or 2.
7. the application of the described molecule marker of claim 6 in the paddy rice cross breeding breeding.
CNB2006100119764A 2006-05-24 2006-05-24 Paddy rice hybrid fertility gene and its application Expired - Fee Related CN100457905C (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108690848A (en) * 2018-02-27 2018-10-23 南京农业大学 Rice interspecific hybrid pollen fertility gene and its application
CN114085847A (en) * 2021-12-22 2022-02-25 云南农业大学 Rice OsSUT3 gene mutant and molecular identification method and application thereof
CN116606916A (en) * 2023-06-30 2023-08-18 河北师范大学 Method for identifying, screening or controlling wheat spike number per spike based on SNP locus

Family Cites Families (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1466627A (en) * 2000-08-17 2004-01-07 �ձ��̲ݲ�ҵ��ʽ���� Method of estimating the genotype of fertility recovery locus to rice BT type male fertility cytoplasm
CN1167331C (en) * 2002-04-27 2004-09-22 华南农业大学 Method for breeding light- and temp.-sensitive nucleus sterile line rice with plant age regulatory fertility

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108690848A (en) * 2018-02-27 2018-10-23 南京农业大学 Rice interspecific hybrid pollen fertility gene and its application
CN114085847A (en) * 2021-12-22 2022-02-25 云南农业大学 Rice OsSUT3 gene mutant and molecular identification method and application thereof
CN116606916A (en) * 2023-06-30 2023-08-18 河北师范大学 Method for identifying, screening or controlling wheat spike number per spike based on SNP locus
CN116606916B (en) * 2023-06-30 2023-11-28 河北师范大学 Method for identifying, screening or controlling wheat spike number per spike based on SNP locus

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