CN107164390A - Rice hybrid pollen fertility gene seat Sc allele Sc j and Sc i separation clone and the application in breeding - Google Patents

Rice hybrid pollen fertility gene seat Sc allele Sc j and Sc i separation clone and the application in breeding Download PDF

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CN107164390A
CN107164390A CN201710476748.2A CN201710476748A CN107164390A CN 107164390 A CN107164390 A CN 107164390A CN 201710476748 A CN201710476748 A CN 201710476748A CN 107164390 A CN107164390 A CN 107164390A
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刘耀光
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    • C12N15/8287—Phenotypically and genetically modified plants via recombinant DNA technology with agronomic (input) traits, e.g. crop yield for fertility modification, e.g. apomixis

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Abstract

The invention belongs to field of plant genetic, rice hybrid pollen fertility gene seat is specifically disclosedScAlleleSc‑ jWithSc‑iSeparation clone and the application in breeding.Japonica riceSc‑jWith long-grained nonglutinous riceSc‑iThe albumen containing DUF1618 domains is encoded, it is required by paddy pollen normal development.Sc‑jOnly one of which gene copy,Sc‑iThere are 2 or 3 tandem gene copies in different kinds.In the hybrid that long-grained nonglutinous rice and japonica rice hybridize,Sc‑iThe high level expression product of multi-copy gene can suppressSc‑jExpression, cause hybrid pollen infertility.The present invention proposes that one kind utilizes gene editing technology revulsionSc‑iCopied part, with reduceSc‑iDosage effect of gene, recover indica rice hybrid inSc‑jThe fertility of pollen, overcomes the cross breeding method of indica-japonica hybrid hybrid male infertility.

Description

Rice hybrid pollen fertility gene seatScAlleleSc-jWithSc-iSeparation gram Application grand and in breeding
Technical field
The present invention relates to field of plant genetic, in particular it relates to rice hybrid pollen fertility gene seatSc's AlleleSc-jWithSc-iSeparation clone and the application in breeding.
Background technology
Plant is hybridized by the hybridization between different cultivars.Hybrid has excellent in terms of growth potential, adaptability, yield Gesture, referred to as hybrid vigour.The yield, quality and resistance of crops can be improved using hybrid vigour.Asian Cultivated Rice is divided into Xian Rice subspecies（Oryza sativa ssp. indica）With japonica rice subspecies（O. sativa ssp. japonica）, between them Hybrid has powerful hybrid vigour, and yield potential is very big.On the other hand, the breeding such as cultivated rice and wild rice hybridization and backcrossing is passed through Method, can will be present in some for example disease-resistant, pest-resistant, high yields of useful character of wild rice etc. and import cultivated rice, cultivate excellent New varieties.But, it is low or sterile to there is fertility in the hybrid that Hybrids of Indica and Japonica and cultivated rice and wild rice hybridization are produced Problem, referred to as hybrid dysgenesis (hybrid sterility).Described hybrid dysgenesis is produced in Oryza species atomization One of mechanism of reproduction isolation, belongs to reproduction after zygote and isolates（postzygotic reproductive isolation）.It is miscellaneous Plant sterile phenomenon and seriously hinder the utilization that rice heterosis is converted into higher rice yield and the useful gene of wild rice.
There are multiple locus in paddy rice（loci）It is relevant with paddy rice hybrid fertility.Described locus is cytogenetics The concept of level, refers to a specific region of chromosome, may include the control homogeneity of the related gene of 1 or more than 1 Shape.In these hybrid fertile genes seat, some are to cause oogamete infertility, such asS5, S7, S8, S9, S15, S17, some It is to cause andro gamete（Pollen）Infertility, such asSa, Sb, Sc, Sd, Se, Sf （Kinoshita 1995, Rice Genet. Newsl., 12:9-15；1988, the Japan J. such as Ikehashi Breed. 38:283-291;Ikehashi etc. 1996, Rice Genet III, pp404-408;1996, the Theor Appl such as Wang Genet 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, heredity Journal 21：34-41).These locus can have different degrees of variation point in different cultivars or strain or wild rice material Change, produce a variety of multiple alleles.According to the rice subspecies type and the effect to hybrid fertile for carrying these multiple alleles Should, they are divided into indica type（S-i）, round-grained rice type（S-j）, neutral type（S-n）Or other transitional types.Described gene symbolS Hybrid fertile gene seat is represented, such asSa, ScEtc. referring to specific hybrid dysgenesis locus,I, j, nRepresent respectively indica type（indica）、 Round-grained rice type（japonica）And neutral type (neutral）Multiple allelic loci, such asSc-i、Sc-jOrSc-n.Neutral type is also referred to as For affinity type（compatibility）Or wide affinity type（wide compatibility）.
In paddy rice hybrid, the hybrid dysgenesis locus of differentiation tends to occur not affine interaction under heterozygous state, causes Carry the andro gamete of a certain allele（Or oogamete）Development is abnormal, and the allele specific of referred to as hybrid kills gamete （allele-specific gamete elimination）Effect.But, these locus are under homozygotic state（Such as homozygosis Parent）The normal development of gamete is not influenceed.The genetic behavior of most of hybrid dysgenesis locus meets " single seat position sporinite-match somebody with somebody The pattern (Oka, 1974, Genetics 77,521-534) of daughter interaction ".For example, with carrying respectivelySc-j/Sc-jGene The paddy rice japonica rice of type andSc-i/Sc-iThe NIL hybridization of genotype, the first generation of hybrid (F of generation1) genotype beSc- j/Sc-i.The andro gamete that its flower pesticide meiosis is produced（Pollen）In, carrySc-iAllele seat it is normal fertile, and carrySc-jThe male gametophyte development of allele is abnormal, therefore shows as about 50% infertility of pollen sterility i.e. half（Fig. 1）.But hybrid is female The fertility of gamete is not influenceed by heterozygous genes seat.As a result the second generation of hybrid is caused（F2）Carried in colonySc-j/Sc-jBase Because the ratio of the plant of type is seriously on the low side, that is, deviateSc-i/Sc-i：Sc-i/Sc-j：Sc-j/Sc-j=1：2：1 Mendel point From rule（Fig. 1）.
The clone of the Hybrid sterility between indica and japonica rice gene of 3 single seat positions is had reported at present, includingS5, Sa, HSA1（Chen etc. 2008,Proc. Natl. Acad. Sci. USA105, 11436-11441；Yang etc. 2012,Science337, 1336-1340；Long etc. 2008,Proc. Natl. Acad. Sci. USA105, 18871-18876;Kubo etc. 2016, Mol. Plant9, 221-232). ScIt is to control one of oligogene seat of rice hybrid pollen fertility（Yang etc. 2004,Chinese Science Bulletin49, 1718-1721）.
The content of the invention
The present invention in breeding of hybridized rice by map-based cloning in order to, efficiently against hybrid dysgenesis sex chromosome mosaicism, be existedScHybrid fertile related gene, including indica type and round-grained rice type allele are cloned in seat；It was found that wherein indica type allele seatSc- iIt is made up of 2 or 3 tandem-repeated copies.On this basis, the present invention proposes that one kind utilizes gene editing technology rite-directed mutagenesisSc-iCopied part, create affinity type alleleSc-n, to recover in hybridSc-jPollen fertility, overcomes hybrid Xian round-grained rice miscellaneous Plant male sterile cross breeding method.
First purpose of the present invention is separation cloning rice hybrid pollen fertility gene seatScDNA sequence dna and its coding Protein sequence.
Second object of the present invention is illustratedScThe molecular genetic mechanism of the function of gene and the infertility that hybridizes.
Third object of the present invention is using its molecular genetic mechanism as according to there is provided create artificial affinity typeSc-nBase Cause, the method for the fertility that crossbreeds is affine system and the application in indica-japonica hybrid breeding.
To achieve these goals, the present invention is achieved by the following technical programs：
Rice hybrid pollen fertility gene seatScControl pollen fertility allele, including single copy equipotential base in japonica rice CauseSc-j, its nucleotide sequence such as SEQ ID NO:Shown in 1；With the tandem-repeated copies allele in long-grained nonglutinous riceSc-iaWithSc-ib,Sc-iaWithSc-ibNucleotide sequence such as SEQ ID NO:2 and SEQ ID NO:Shown in 3.SEQ ID NO:1 institute Show in sequence, 1~1000bp be promoter sequence, 2285~3678bp be intron sequences, 1001~2284bp and 3679~ 3735bp is exon sequence.SEQ ID NO:In sequence shown in 2,1~1000bp is promoter sequence, and 2276~3673bp is Intron sequences, 1001~2275bp and 3674~3730bp are exon sequence.SEQ ID NO:In sequence shown in 3,1~ 1000bp is promoter sequence, and 2276~3669bp is intron sequences, and 1001~2275bp and 3670~3726bp are outer aobvious Subsequence.
AlleleSc-jThe protein sequence of coding such as SEQ ID NO:Shown in 4, alleleSc-iaThe albumen of coding Sequence such as SEQ ID NO:5, alleleSc-ibThe protein sequence of coding such as SEQ ID NO:Shown in 6.
Present invention discover that japonica riceSc-jWith long-grained nonglutinous riceSc-iThe albumen containing DUF1618 domains is encoded, is paddy rice Powder normal development institute is required.Sc-jOnly one of which gene copy,Sc-iCopied in different kinds with 2 or 3 tandem genes Shellfish.
It has also been found that in long-grained nonglutinous rice and japonica rice hybrid,Sc-iThe high level expression product of multi-copy gene can suppressSc-jExpression, cause hybrid pollen infertility.Therefore, the present invention establishes a kind of side for cultivating the affine system of paddy rice hybrid fertility Method, is concretely comprised the following steps, and will be contained using genome editing techniqueSc-iIn the rice varieties of alleleSc-iaOr/andSc- ibPortion gene copies rite-directed mutagenesis, reductionSc-iThe active gene copy number of locus, hybridize affine system.As optimal Embodiment, a kind of method of the affine system of cultivation paddy rice hybrid fertility concretely comprised the following steps, will be contained using genome editing technique HaveSc-iIn the rice varieties of alleleSc-iaOr/andSc-ibPortion gene copies rite-directed mutagenesis, last only to retain one Individual Sc-ia gene copies, or only retain oneSc-ib, so as to reduceSc-iThe active gene copy number of locus, hybridizes Affine system.
BecauseSc-iThere is 2 or 3 tandem genes copy in different kinds, thus for containingSc-ia、Sc-ib1 WithSc-ib2Rice varieties, utilize genome editing technique willSc-ia、Sc-ib1OrSc-ib2In any one or it is any Two gene copies carry out rite-directed mutagenesis, reductionSc-iThe active gene copy number of locus, hybridize affine system, whereinSc-ib1WithSc-ib2Nucleotide sequence it is sameSc-ib,Sc-ib1WithSc-ib2It isSc-ibTwo gene copies name. Plant CRISPR/Cas9 gene editing systems are utilized as preferred embodiment, concretely comprising the following steps（Ma etc. 2015,Mol. Plant8, 1274-1284）, to long-grained nonglutinous riceSc-iRegion is directed toSc-ib1 WithSc-ib2Code area carry out specific mutations, but RetainSc-iaNormal function, the mutation of these portion genes can reduceSc-iActive gene copy number, and a remaining gene Copy（Sc-ia）Expression can maintain its function to pollen fertility.
For containingSc-ib1WithSc-ib2Rice varieties, utilize genome editing technique willSc-ib1OrSc-ib2In Any one gene copy carry out rite-directed mutagenesis, reduction reductionSc-iThe active gene copy number of locus, hybridize parent Be；Sc-ib1WithSc-ib2Nucleotide sequence it is identical, beSc-ibTwo gene copies.Preferably, it is of the present invention Genome editing technique is CRISPR/Cas9 gene editing systems, it is not limited to this gene editing technology.
As a kind of preferred embodiment, using CRISPR/Cas9 gene editings technology by long-grained nonglutinous riceSc-iSeatSc- ib1OrSc-ib2In any one gene copy carry out rite-directed mutagenesis specific method be,
S1. utilizeSc-ibThe G nucleotide variations that gene coding region is the 235th, design one is rightSc-ibThe CRISPR/ of gene specific Cas9 gene editing target spots, this special target spot sgRNA of construction expression CRISPR/Cas9 carriers；
S2. contained with the conversion of this genetic modification carrierSc-iThe rice varieties of allele, are obtainedSc-ibAllele-specific Mutation, andSc-iaAllele is the mutant of wild type.
Application of the method for the present invention for cultivating the affine system of paddy rice hybrid fertility in paddy rice cross breeding breeding, specifically should It is that methods described is cultivated to the obtained affine system of paddy rice hybrid fertility and japonica rice variety hybridization with method, cultivatesScSeat is not produced The hybrid of pollen abortion, for paddy rice cross breeding breeding.
It should be appreciated that being existed based on gene editing technologies such as programmable nuclease such as TALLENs and CRISPR/Cas9/Cpf1 The application of plant especially paddy rice is quite ripe（2016, the Molecular such as Ma Plant, 10.1016/ j.molp.2016.04.009）.Gene order, gene function and allele expression and the hybrid provided in the present invention On the basis of the information such as relation of infertility, those skilled in the art can to these gene selects 1 or several be different from this hair The target spot of bright implementation carries out gene elmination or base mutation, easily reaches the same or analogous hybrid compatibility effect of the present invention Really.Therefore, the present invention includes any fragment deletion to these genes and its promoter regulation area and any site of code area Editor mutation.
Compared with prior art, the present invention has the advantages that：
The hybrid fertile gene and its affine system's creation method that the present invention is provided have important application value.The inventive method can With the affine system of quickly breeding hybrid fertile, these are affine, and system can also cultivate new with other rice varieties or incross Hybrid fertile is affine system.It is that, for Rice Cross Breeding Between Indica And Japonica, can overcome that these hybrid fertiles are affineScHybrid dysgenesis, improves hybrid Fertility, reach the purpose of volume increase.
Brief description of the drawings
Fig. 1 shows paddy riceScEffect of the hereditary pattern and its genotype of gene to pollen fertility.Japonica rice T65 waits base near Because being the normal fertile pollen of E5 generations, its hybrid F1Produce the pollen sterility of about half（Arrow refers to abortive pollen）, and F2 The genotype separation substantial deviation 1 that colony produces:2:1, whereinSc-jHomozygotic number seldom, show containingSc-jPollen Abortion.
Fig. 2 is paddy riceScFinely positioning and gene order structure of the locus in the 3rd chromosome；The number that figure (A) is respectively marked Word is represented in the position disclosed in one on bacterial artificial chromosome (BAC) simultaneously（kb）；Scheme the gene order-checking of (B) display Analysis shows,ScLocus only has 1 gene in japonica riceOs03g0247300, it is named asSc-j;Different rice varietiesSc Locus contains 2-3Os03g0247300Homologous tandem repeat locus, is named asSc-ia, Sc-ib1, Sc-ib2；Sc-is The pseudogene of destruction is inserted for transposons；Sc-ia, Sc-ib1WithSc-ib2Promoter and upstream sequence withSc-jStartup Son and upstream sequence are entirely different；Sc-j,Sc-ia,WithSc-ib （Sc-ib1WithSc-ib2）Sequence such as SEQ ID NO: 1~3 It is shown.
Fig. 3 isSc-jFunctional study；Figure（A）To incite somebody to actionSc-jAntisense expression vector converts T65, and the transgenosis of acquisition is hemizygous Plant（T1Generation）,Sc-jIn the two cores early stage of its peak expression（early-bicellular stage）Expression decline； Figure（B）For antisense transgene T1The hemizygous plant performance pollen semi-sterility of transgenosis in generation（Arrow refers to abortive pollen）；Relative expression It is worth for reference geneOsActin 1Ratio.
Fig. 4 is that quantitative RT-PCR detects japonica rice T65, NIL E5, and its hybrid F1'sScGene expression；Scheme (A) WithScAllele universal primer（P1, P2）The expression of different development stage flower pesticide is detected, as a result shows T65'sSc-j Mainly in two core early stage specifically expressings；Figure（B）According toSc-jSpecial 5 ' non-translational region designSc-jSpecial primer（P3, P4）Hybrid F is compared in detection1With T65 two core early stage flower pesticide often copySc-jExpression, as a result show F1'sSc-jTable It is significantly less than T65's up to levelSc-jExpression；Relative Expression values for pairOsActin 1Ratio.
Fig. 5 is to be modified with CRISPR/Cas9 gene editings systemSc-iPart repeat copy；According toSc-ib1WithSc- ib1Code area the 235th G nucleotide variations formation TGG be PAM elements（It is that the identification of Cas9-sgRNA complexs combines target spot The required element of sequence）, design one it is rightSc-ib/Sc-ib1The CRISPR/Cas9 gene editing target spots of gene specific（It is lower to draw Line）；E5-ed1 and E5-ed2 is specific mutagenesisSc-ib1With Sc-ib2The affine system of gene editing.
Fig. 6 is to overcome hybrid sterility with the affine system of gene editing；Figure（A）For with E5-ed1 and E5-ed2 and japonica rice T65 Hybridization, obtains the gene editing hybrid F that pollen fertility is improved1（Arrow refers to abortive pollen）；Figure（B）ForSc-ibGene editing is miscellaneous Plant F2In colonySc-jHomozygous individual number increase, its genotype separation meets 1:2:1 (ns represents that difference is not notable,jj, ij, iiRepresent respectivelySc-jHomozygote, heterozygote, andSc-iHomozygote, including wild type andSc-ibEdit saltant type)；Figure（C）It is aobvious Show gene editing hybridSc-jExpression is improved largely compared with normal hybrid.
Embodiment
The present invention is made with reference to Figure of description and specific embodiment and further being elaborated, the embodiment It is served only for explaining the present invention, is not intended to limit the scope of the present invention.Test method used in following embodiments is such as without spy Different explanation, the conventional meanses being well known to the skilled person and the science of heredity and molecular biology manipulations bar using standard Part.
Embodiment 1： ScFinely positioning clone and Sequence structure analysis
The early-stage Study of the present inventor willScIt is positioned at approximate region (Yang etc. 2004. on the chromosome of paddy rice the 3rdChinese Science Bulletin49, 1718-1721).The present invention is further with map based cloning method as shown in Figure 2 CloneScGene.According to the data (http in disclosed Public Rice Genome Sequence Data storehouse://www.ncbi.nlm.nih.gov), In the regional development multiple molecular labelings (tables 1).With T65 (Sc-j/Sc-jGenotype) and its NIL E5 （Sc-i/Sc-i) hybridization, produce F1And F2Colony, by 8689 plants of F of linkage analysis2Individual,ScFinely positioning is in one, japonica rice 8.6 kb interval（Fig. 2A）.According to Public Rice Genome Sequence Data storehouse GenBank (http:// Www.ncbi.nlm.nih.gov), 1 numbering of note is in the kb of japonica rice 8.6 intervals of positioningOs03g0247300Base Cause, the present invention using it asSc-jGene, its nucleotide sequence such as SEQ ID NO:1, coding has SEQ ID NO:Shown in 4 Amino acid polypeptide sequence, contain a DUF1618 conserved domain.
According to Public Rice Genome Sequence Data storehouse GenBank (http://www.ncbi.nlm.nih.gov), in long-grained nonglutinous rice Kind 93-11'sOs03g0247300There are larger structure and sequence variations in equipotential interval.But it is due to the 93-11 interval Sequence is incomplete, present invention PCR（PCR）Technology（Using TOYOBO KOD FX archaeal dna polymerases, by product Specification imposes a condition operation）Amplified from 93-11 and rice variety bright extensive 63 and NIL E5Sc-iInterval base Because of pack section and determine its sequence.As a result show, E5'sOs03g0247300Correspondence gene（Containing withSc-jHomologous promoter Sequence）Code area there is the insertion of 2 transposons, therefore be the pseudogene of an inactivation（Sc-is）.Sc-isDownstream has 3 Individual is respectively 28.2 and 28.5 kb forward repetitive sequence, and each repetitive sequence contains oneOs03g0247300Homologous gene, But their promoter and upstream sequence（Common 23.9kb）WithSc-jPromoter and upstream sequence it is entirely different.The present invention by this The recombination of repetitive sequence is named asSc-ia, Sc-ib1WithSc-ib2（Fig. 2 B）.Sc-iaNucleotide sequence such as SEQ ID NO:2, coding has SEQ ID NO:Amino acid polypeptide sequence shown in 5；Sc-ib1 WithSc-ib2With identical nucleosides Acid sequence such as SEQ ID NO:3, coding has SEQ ID NO:Amino acid polypeptide sequence shown in 6.93-11 and bright extensive 63 contains HaveSc-is,Sc-ib1 WithSc-ib2Copy, but do not haveSc-iaCopy.These discoveries show, long-grained nonglutinous riceSc-iSeat there occurs Big structure variation and gene copy number variation, is 1 complex locus, and japonica riceSc-jIt is 1 simple locus.Sequence Compare and show,Sc-j,Sc-ia, Sc-ib1(Sc-ib2) code area between there is the variation of 19 single bases and 33 bases are inserted Enter deletion mutation.
Primer sequence used in the present invention of table 1
Embodiment 2： ScThe functional verification of gene
Expanded with RT-PCR technology from japonica riceSc-jCDNA fragments (are located at the bp of code area the 514th -1284), with Opposite direction connection To cornUbiquitinGene promoter is connected, and is built into antisense gene expression binary vector.T65 is converted with this carrier, obtained Transformant.Transgenosis hemizygote（1 chromosome has another 1 homologue of transgenosis not have transgenosis）'sSc-jExpression Reduction（Fig. 3 A）, there is the pollen sterility of substantially half（Fig. 3 B）.ProveSc-jIt is pollen gametophyte fertile gene.
Embodiment 3：ScHybrid dysgenesis Analysis on Mechanism
With primer P3 and P8, (amplification is allScAllele) quantitative RT-PCR is carried out, to T65, E5, hybrid F1Expression analysis Show,Sc-jMain to be expressed in two core early stage pollen-specifics, expression is relatively low.Sc-iDuplicate factor is female thin in flower pesticide microspore Born of the same parents express to the different developmental phases of two core early stage pollen, and expression is higher（Fig. 4 A）.WithSc-jSpecial primer（P3, P4）Carry out quantitative RT-PCR expression analysis to show, F1'sSc-jThe expression each copied is only equivalent to its parent T65 pact 1/5（Fig. 4）, show in hybridSc-jExpression be suppressed and produce pollen abortion.
Embodiment 4：The creation method of hybrid is affine system
In order to illustrate in hybridSc-jExpression inhibiting whether withSc-iThe high level expression of multiple copies is relevant, and exploitation wound Build the new method of the affine system of hybrid, basis of the present inventionSc-ib1WithSc-ib1The G nucleotide variations of code area the 235th formed TGG is the required element PAM of CRISPR/Cas9 gene editings, and design one is rightSc-ib1/Sc-ib2Gene specific CRISPR/Cas9 gene editing target spots（5’-GCGGAAGGCGATCAGGGTAG-3’）, compiled using plant CRISPR/Cas9 genes Collect system and its carrier construction method（Ma etc. 2015,Mol. Plant8, 1274-1284）, construct 1 CRISPR/Cas9 Genetic modification carrier.Transformant is obtained with this carrier Transformed E 5, is sequenced, reflected through fragment of the PCR amplifications containing target sequence Surely arriveSc-ib1WithSc-ib2Undergo mutation（Sc-iaFor wild type）Specific mutants E5-ed1 and E5-ed2（Fig. 5）.
Embodiment 5：The application of hybrid is affine system
The affine system of hybrid and the japonica rice hybridization that above-described embodiment 4 is obtained obtain F1Hybrid.Observation shows, these F1Hybrid can Recover pollen fertility in various degree（Fig. 6 A）.F2Genotype separates analysis shows, each colonySc-jHomozygotic ratio is significantly Degree rises, and the segregation ratio of its colony meets 1:2:1（Fig. 6 B）.WithSc-jSpecial primer（P1, P2）Carry out quantitative RT-PCR expression Analysis shows, these gene editings are affine, and system combines F1HybridSc-jExpression is than wild type F1Significantly improve（Fig. 6 C）. Therefore, the affine system of gene editing hybrid that the present invention is created has compatibility gene typeSc-n, can be used in paddy rice cross breeding breeding OvercomeScHybrid dysgenesis problem.
SEQUENCE LISTING
<110>Agricultural University Of South China
<120>Rice hybrid pollen fertility gene seat Sc allele Sc-j and Sc-i separation are cloned and in breeding
Using
<130>
<160> 6
<170> PatentIn version 3.3
<210> 1
<211> 3735
<212> DNA
<213>Sc-j genes
<400> 1
aaatgggcta agtattttat tcacaagtga aacgcccgag actaatgtaa attagtgtta 60
gtatagtaac tttaggcttg tcgattagtg cgtgaaaagc aatagttaga taaatttggg 120
tttactagac taatctttgc gccacacaat catttgcatg tcaaaactac ctataaaacc 180
actgtagggg ttatttgaac cgattttcat acttcaaagg tagaaatcta gtttcgtact 240
ttataagtgg ttaaatagac caaccccaac acttcagggt attccttata tctactatag 300
ttttaacttt gagttaaatc ttaaggtggt ccatgcccta cggcagtagc ggagccagaa 360
atttaaatct gtggggtcaa ttactagtgg tttgtattta agtgggatca tttctcttta 420
ttgttttgga tttatacata aaaattgagg tttataggac gttttttaaa aagctggggt 480
cggccgaccc cacagaaatg gcctaaatcc accactgccc caggtgccac ctagttaact 540
ctgcatatgg gcctatgatc ccttcgatca cttttcatta ctacaatggt gtttgaaact 600
gatggatatt aagctggtgc atgtagaatg aagaaaataa ttaacacatt attgattaaa 660
atttaattat taaaaacttg acaaattaat ttatttgata attttaatgt aacttttata 720
tagaaagttt gaaaagtgta ataacggaaa ctgagttaaa atctgtacgt gttaaatata 780
ttattcccct ttttaaattt atagaagaag ttaaatatga acgttggaaa acgctgaaca 840
agtgaacata tcccgcgatc cgcatttgtt gcgactcgtt tcgatcaact gattctcctt 900
cccggccgga catccacatc atcctgcgat cgttcgtttc cctagtcgtt ccaggctttt 960
tcagctctac tcccctccac ccaggcgcaa atcatccgcc atggctcccg cctgggtctt 1020
gctcgatcgc gtcgtcaagc ccgccgtctt cgacgaagaa gagagcaaag gtaagggaga 1080
atcgaccggc gccccagtga agtacctgcc agccagatta aggcaggaag tccccgccgg 1140
catgcgggac gtgaagccct acccagaggt tgcggatcct cccattatat ctcgcttctc 1200
catgctgatt tcgcggaagg cgatcagggt agtgaaatcc gtccgtgtcc ggtgcgccga 1260
caagagcctt gtcttgttct acgccggcac cggcttcccc ggcttctcgt cccacggctg 1320
ccacctgatc tacgacgcca tcgacggctc tctcactgcg gtgcacacat tcccctttcc 1380
tgtgtccgga gtcgtctggg tcggcagagc cgctgtcctg cgccatgccg gcggcggagg 1440
aggaggagga gacggtacta ccgcctccta tgtcatcgcc gagctgctca ggccgtttca 1500
tggctccctt cccgatgcca cgctcgtgat gtggttgtcg aattcccccg cgtcgacctc 1560
cggcagcaac ggccaatggg tgaaggagga cgttcgcctt cccggcgagg tgtgcacggg 1620
caccgacccc ttcaccaccg acttggtgtt ctcgttcggt gaatcgtgcc tgtgctgggc 1680
cgatctgttc atgggcatcc tgttctgcga ccttgcgacg ctgcgtgcac ctcggttccg 1740
tttcatccca ttgcccaagg cttgctcctt cgaccccgtc ggcaagtatg gccggcctca 1800
catgcccgag ttccgttcca tgggccgcgt caacggtgtc atcaggttga tcgacatgga 1860
gggttttacc aacgagtacc tggccgtgga tgaggtgaag ctgaccatct ggaccctgtc 1920
agccgacctc agcgagtggg agaaaggccc ggtgtgtact gtaggagaca tttgggccag 1980
tgaggaattc gtcgccatgg ggttgccgca gcttagaccg atgtgccctg tcctgagcat 2040
ggttgacgaa gatgtcgtct gtgtagtcat gactgaagtc gagattgagg agagcgatgt 2100
cacagatttc gacgatgagg gcaacaagct caagttcaag gcccagtacg ttcttgacat 2160
cgacgtgcgg cgtaagcggg tgttatccat cacccagcac cacattgaaa gtatgggtga 2220
tcttatacca gatctcatcg cctgtgagtt cactgcatac tcggagctct caaaaggcat 2280
gcaggtatta ctactacgac acccaacata cttagttatt tactcacttg aagtatattg 2340
ttctgttaac acaataacac tttccacagt tctaatgctt ggttatcatg tccagtcagt 2400
tacatcttag tatctcacga gtaacggagt aagtatatta ggctagttat cctactgaac 2460
tgaactctag atagcatatg tgtgtgctaa tcagccatca tgattgtcta atgccaaatg 2520
acttagctct tttgccatgc tacagccctc ctcgtccaaa ccaggaacgc cattttttca 2580
ttctgttggt acttctagga actggagaac gctgtcatag ttactttcac ttcactacaa 2640
agttgtgatg gcttgtccag tagatgtatc caattgaatt cttgattcag aattgatata 2700
gacctatttt atttaaagat gttacatgtg gtaaattcaa actgagcacc aagtcaccag 2760
caaggttctg taatgaaatt tgtgaaaagt gcaatctatc cataacgtgt gggcttggtt 2820
ggttttagct attttactta aatgcctctt agaattgacc ttatcagcat gcccttgtgt 2880
gagtattgct cattattatt ttagagagta cagacaaatg gggccaatga taggtatgtt 2940
gcggcgaggc aactacatca tctgaccaaa cctgaccata attcttgtcg cagttgtacg 3000
aaaatctctt ttctttttta cttctgaagt ttttttagcg ttgtgtgttg gaaaagagat 3060
ttgtctaact aaaattctct atgtaataag tatagaatcg tatcctctat accatagtat 3120
atggaaatat catgtgtacc agtgtaaggc cctaactgag ttatgttact ttccctagct 3180
ggtactccct ccatttcaca ttataagact ttctagtatt gcacacattc atatatatgt 3240
taatgaatct aaacatatac tccctccgtt ttgaaatgtt tgacaccgtt gactttttag 3300
cacatgtttg accattcgtc ttattcaaaa acttttgtga aatatgtaaa attatatgcc 3360
tacataaaaa tatatttaac aatgaatcaa atgataggaa aataattaat aattacttaa 3420
attttttgaa taagacgaac ggtcaaacat gtgctaaaaa gtcaacggcg tcaaatatgt 3480
gtagtagatg tcctaagact atctaggttt ctcaatgatg aattttcatt tatatatatt 3540
tggccacata ttaatttcct gtctagctct gttaattgta actttggtat gcactgaggg 3600
aattattgaa gtggattaag aaccattagg ccattttaaa tttgtctagt ttgacataca 3660
ttaatttgtt tctgctaggc aatggtggaa ggaaatgagg gtgaggaaag cacaaagcgg 3720
atgaaggtta aatga 3735
<210> 2
<211> 3730
<212> DNA
<213>Sc-ia genes
<400> 2
gttgactagt gttgtagatc cagtatagca aaagaccaag aattaaaaca acggcttaaa 60
aacagcacaa acagaatgct aaaaggttgc aaaccagtaa gcaagtataa tggacgacac 120
attttgttcg gatcatcacc tctttctcct tatccatcag aaaacaccga gacagcgaaa 180
ggtcgacgag cccatccttg gaagtggacg ccaactccgt agccgccact gcagcctcgt 240
gcgaggcagc cgctgatgta ggcgccggcg gcacttgatc gtccacgatc agcttctgcc 300
tcagatcttt cttctccagc gttgctgccg gttccaacga cgatggaacc gccatgggag 360
ctagtaagga caccaccatc ggacgcgaat cagaacagcg gcagccggcg cttgacgtgc 420
cgcgcccgtg agcaccgagc ccgtcggcgt cgcctcggtc cctgccgcgg cagcggctgc 480
ggctgcggcg tcgacggatc attagggtgc ggaaggagtg agagagcgga cagcggagga 540
acaaggtgct gtctctcacc cttccttcca ctgcacctat agccttttca tctcctgcac 600
ctatgcctag tccacaagac agaggcgtcc tcctctcctc tctctcctct ctcctctcac 660
cggtgcataa tgaacctagt ctacaaacta ggctcccttt gtgtttgttg cgggtggcat 720
atccaccgtc cattagcctc tctatctccg ccgccatatc caccgtccat tagtctcccg 780
atccccgcca ctctaccttt gatggggccc gtttccgatc tggactcgtc ttctccctct 840
cgacggcaca gtgggatctg ggaagaggga agggtttgct cgatcccttt gttcttgccg 900
tctcttgcac tgatctagct aattctttct gctggggcac tcgaaaaaaa actcttcgac 960
tgctccccgt actccaaacc cacccacgcg caaatccgcg atggctccca cctgggtctt 1020
gctcgatcgc ttcgtcaagc ccactatctt cgacgaagaa gagagcaaag gtaagggaga 1080
atcgaccggc gccccagtga agtaccttcc agccagatta aggcaggaag tccccgccgg 1140
catgcgggac gtgaagccct acccagaggt tgcggatcct cccattgtat ctcgcttctc 1200
catgctgatt tcgcggaagg cgatcagggt agtgaaatcc gtccgtgtcg agtgcgccga 1260
caagagcctt gtcttgttct acgccggcac cggcttcccc ggcttctccc acggctgcca 1320
cctgatctac gacgccatcg acggttctct cactgcagtg cacacgttcc cctttcctgt 1380
gtccggagtc gtctgggtcg gcacagccgc tgtcctgcgc catgccggcg gcggcggagg 1440
aggagacggt actaccgcct atgtcatcgc cgagctgctc aggccgttcc atggctccct 1500
tcccgatgcc acgctcgtga tgtggttgtc gaattccccc gcgtcgacct ccggcagcaa 1560
tggccaatgg gtgaaggagg acgttcgcct tcccggcgag gtgtgcacgg gcaccgaccc 1620
cttcaccacc gacttggtgt tctcgttcgg tgaatcgtgc ctgtgctggg ccgatctgtt 1680
catgggcatc ctgttctgcg accttgcgac gctgcgtgca cctcggttcc gtttcatccc 1740
attgcccaag gcttgctcct tcgaccccgt cggcaagtat ggccggcctc acatgcccga 1800
gttccgttcc atgggccgcg tcaacggtgt catcaggttg atcgacatgg agggttttac 1860
caacgagtac ctggccgtgg atgaggtgaa gctgaccatc tggaccctgt cagccgacct 1920
cagcgagtgg gagaaaggcc cggtgtgtac tgtaggagac atttgggcca gtgaggaatt 1980
cgtcgccatg gggttgccgc agcttagacc gatgtgccct atcctgagca tggttgacga 2040
agatgtcgtc tgtgtagtca tgactgaagt cgagattgag gagagcgatg tcacagattt 2100
cgacgatgag ggcaacaagc tcaagttcaa ggcccagtac gttcttgaca tcgacgtgcg 2160
gcgtaagcgg gtgttatcca tcacccagca ccacattgaa agtatgggtg atcttatacc 2220
agatctcatc gcctgtgagt tcactgcata ctcggagctc tcaaaaggca tgcaggtatt 2280
actactacga cacccaacat acttagttat ttactcactt gaagtatata tattgttctg 2340
ttaacacaat aacactttcc acagttctaa tgcttggtta tcatgtccag tcagttacat 2400
cttagtatct cacgagtaac ggagtaagta tattaggcta gttatcctac tgaactgaac 2460
tctagatagc atatgtgtgt gctaatcagc catcatgatt gtctaatgcc aaatgactta 2520
gctcttttgc catgctacag ccctcctcgt ccaaaccagg aacgccattt ttgcattctg 2580
ttggtacttc taggaactgg agaacgctgt catagttact ttcacttcac tacaaagttg 2640
tgatggcttg tccagtagat gtatccaatt gaattcttga ttcagaattg atatagacct 2700
attttattta aagatgttac atgtggtaaa ttcaaactga gcaccaagtc accagcaagg 2760
ttctgtaatg aaatttgtga aaagtgcaat ctatccataa cgtgtgggct tggttgattt 2820
tagctatttt acttaaatgc ctcttagaat tgaccttatc agcatgccct tgtgtgagta 2880
ttgctcatta ttattttaga gagtacagac aaatggggcc aatgataggt atgttgcggc 2940
gaggcaacta catcatctga ccaaacctga ccataattct tgtcgcagtt gtacgaaaat 3000
ctcttttctt ttttacttct gaagtttttt tagcgttgtg tgttggaaaa gagatttgtc 3060
taactaaaat tctctatgta ataagtatag aatcgtatcc tctataccat agtatatgga 3120
aatatcatgt gtaacagtgt aaggccctaa ctgagttatg ttactttccc tagctggtac 3180
tccctccatt tcatattata agactttcta ttattgcaca cattcatata tatgttaatg 3240
aatctaaaca tatactccct ccgttttgaa atgtttgaca ccgttgactt tttagcacat 3300
gtttgatcat tcgtcttatt caaaaacttt tgtgaaatat gtaaaattat atgcctacat 3360
aaaaatatat ttaacaatga atcaaatgat aggaaaataa ttaataatta cttaaatttt 3420
ttgaataaga cgaacggtca aacatgtgct aaaaagtcaa cggcgtcaaa tatgtgtagt 3480
agatgtccta agactatcta ggtttctcaa tgatgaattt tcatttatat atatttggcc 3540
acatattaat ttcctgtcta gctctgttaa ttgtaacttt ggtatgcact gagggaatta 3600
ttgaagtgga ttaagaacca ttaggccatt ttaaatttgt ctagtttgac atacattaat 3660
ttgtttctgc taggcaatgg tggaaggaaa tgagggtgag gaaagcacaa agcggatgaa 3720
ggttaaatga 3730
<210> 3
<211> 3726
<212> DNA
<213>Sc-ib1 or Sc-ib2 genes
<400> 3
gttgactagt gttgtagatc cagtatagca aaagaccaag aattaaaaca acggcttaaa 60
aacagcacaa acagaatgct aaaaggttgc aaaccagtaa gcaagtataa tggacgacac 120
attttgttcg gatcatcacc tctttctcct tatccatcag aaaacaccga gacagcgaaa 180
ggtcgacgag cccatccttg gaagtggacg ccaactccgt agccgccact gcagcctcgt 240
gcgaggcagc cgctgatgta ggcgccggcg gcacttgatc gtccacgatc agcttctgcc 300
tcagatcttt cttctccagc gttgctgccg gttccaacga cgatggaacc gccatgggag 360
ctagtaagga caccaccatc ggacgcgaat cagaacagcg gcagccggcg cttgacgtgc 420
cgcgcccgtg agcaccgagc ccgtcggcgt cgcctcggtc cctgccgcgg cagcggctgc 480
ggctgcggcg tcgacggatc attagggtgc ggaaggagtg agagagcgga cagcggagga 540
acaaggtgct gtctctcacc cttccttcca ctgcacctat agccttttca tctcctgcac 600
ctatgcctag tccacaagac agaggcgtcc tcctctcctc tctctcctct ctcctctcac 660
cggtgcataa tgaacctagt ctacaaacta ggctcccttt gtgtttgttg cgggtggcat 720
atccaccgtc cattagcctc tctatctccg ccgccatatc caccgtccat tagtctcccg 780
atccccgcca ctctaccttt gatggggccc gtttccgatc tggactcgtc ttctccctct 840
cgacggcaca gtgggatctg ggaagaggga agggtttgct cgatcccttt gttcttgccg 900
tctcttgcac tgatctagct aattctttct gctggggcac tcgaaaaaaa actcttcgac 960
tgctccccgt actccaaacc cacccacgcg caaatccgcg atggctccca cctgggtctt 1020
gctcgatcgc ttcgtcaagc ccactatctt cgacgaagaa gagagcaaag gtaagggaga 1080
atcgaccggc gccccagtga agtaccttcc agccagatta aggcaggaag tccccgccgg 1140
catgcgggac gtgaagccct acccagaggt tgcggatcct cccattgtat ctcgcttctc 1200
catgctgatt tcgcggaagg cgatcagggt agtggaatcc atccatgtcg agtgcgccga 1260
caagagcctt gtcttgttct acgccggcac cggcttcccc ggcttctccc acggctgcca 1320
cctgatctac gacgccatcg acggttctct cactgcagtg cacacgttcc cctttcctgt 1380
gtccggagtc gtctgggtcg gcacagccgc tgtcctgcgc catgccggcg gcggcggagg 1440
aggagacggt actaccgcct atgtcatcgc cgagctgctc aggccgttcc atggctccct 1500
tcccgatgcc acgctcgtga tgtggttgtc gaattccccc gcgtcgacct ccggcagcaa 1560
tggccaatgg gtgaaggagg acgttcgcct tcccggcgag gtgtgcacgg gcaccgaccc 1620
cttcaccacc gacttggtgt tctcgttcgg tgaatcgtgc ctgtgctggg ccgatctgtt 1680
catgggcatc ctgttctgcg accttgcgac gctgcgtgca cctcggttcc gtttcatccc 1740
attgcccaag gcttgctcct tcgaccccgt cggcaagtat ggccggcctc acatgcccga 1800
gttccgttcc atgggccgcg tcaacggtgt catcaggttg atcgacatgg agggttttac 1860
caacgagtac ctggccgtgg atgaggtgaa gctgaccatc tggaccctgt cagccgacct 1920
cagcgagtgg gagaaaggcc cggtgtgtac tgtaggagac atttgggcca gtgaggaatt 1980
cgtcgccatg gggttgccgc agcttagacc gatgtgccct gtcctgagca tggttgacga 2040
agatgtcgtc tgtgtagtca tgactgaagt cgagattgag gagagcgatg tcacagattt 2100
cgacgatgag ggcaacaagc tcaagttcaa ggcccagtat gttcttgaca tcgacgtgcg 2160
gcgtaagcgg gtgttatcca tcacccagca ccacattgaa agtatgggtg atcttatacc 2220
agatctcatc gcctgtgagt tcactgcata cttggagctc tcaaaaggca tgcaggtatt 2280
actactacga cacccaacat acttagttat ttactcactt gaagtatatt gttctgttaa 2340
cacaataaca ctttccacag ttctaatgct tggttatcat gtccagtcag ttacatctta 2400
gtatctcacg agtaacggag taagtatatt aggctagtta tcctactgaa ctgaactcta 2460
gatagcatat gtgtgtgcta atcagccatc atgattgtct aatgccaaat gacttagctc 2520
ttttgccatg ctacagccct cctcgtccaa accaggaacg ccatttttgc attctgttgg 2580
tacttctagg aactggagaa cgctgtcata gttactttca cttcactaca aagttgtgat 2640
ggcttgtcca gtagatgtat ccaattgaat tcttgattca gaattgatat agacctattt 2700
tatttaaaga tgttacatgt ggtaaattca aactgagcac caagtcacca gcaaggttct 2760
gtaatgaaat ttgtgaaaag tgcaatctat ccataacgtg tgggcttggt tgattttagc 2820
tattttactt aaatgcctct tagaattgac cttatcagca tgcccttgtg tgagtattgc 2880
tcattattat tttagagagt acagacaaat ggggccaatg ataggtatgt tgcggcgagg 2940
caactacatc atctgaccaa acctgaccat aattcttgtc acagttgtac gaaaatctct 3000
tttctttttt acttctgaag tttttttagc gttgtgtgtt ggaaaagaga tttgtctaac 3060
taaaattctc tatgtaataa gtatagaatc gtatcctcta taccatagta tatggaaata 3120
tcatgtgtaa cggtgtaagg ccctaactga gttatgttac ttttcctagc tggtactccc 3180
tccatttcac attataagac tttctagtat tgcacacatt catatatatg ttaatgaatc 3240
taaacatata ctccctccgt tttgaaatgt ttgacaccgt tgacttttta gcacatgttt 3300
gatcattcgt cttattcaaa aacttttgtg aaatatgtaa aattatatgc ctacataaaa 3360
atatatttaa caatgaatca aatgatagga aaataattaa taattactta aattttttga 3420
ataagacgaa cggtcaaaca tgtgctaaaa agtcaacggc gtcaaatatg tgtagtagat 3480
gtcctaagac tatctaggtt tctcaatgat gaattttcat ttatatatat ttggccacat 3540
attaatttcc tgtctagctc tgttaattgt aactttggta tgcactgagg gaattattga 3600
agtggattaa gaaccattag gccattttaa atttgtctag tttgacatac attaatttgt 3660
ttctgctagg caatggtgga aggaaatgag ggtgaggaaa gcacaaagcg gatgaaggtt 3720
aaatga 3726
<210> 4
<211> 446
<212> PRT
<213>Sc-j albumen
<400> 4
Met Ala Pro Ala Trp Val Leu Leu Asp Arg Val Val Lys Pro Ala Val
1 5 10 15
Phe Asp Glu Glu Glu Ser Lys Gly Lys Gly Glu Ser Thr Gly Ala Pro
20 25 30
Val Lys Tyr Leu Pro Ala Arg Leu Arg Gln Glu Val Pro Ala Gly Met
35 40 45
Arg Asp Val Lys Pro Tyr Pro Glu Val Ala Asp Pro Pro Ile Ile Ser
50 55 60
Arg Phe Ser Met Leu Ile Ser Arg Lys Ala Ile Arg Val Val Lys Ser
65 70 75 80
Val Arg Val Arg Cys Ala Asp Lys Ser Leu Val Leu Phe Tyr Ala Gly
85 90 95
Thr Gly Phe Pro Gly Phe Ser Ser His Gly Cys His Leu Ile Tyr Asp
100 105 110
Ala Ile Asp Gly Ser Leu Thr Ala Val His Thr Phe Pro Phe Pro Val
115 120 125
Ser Gly Val Val Trp Val Gly Arg Ala Ala Val Leu Arg His Ala Gly
130 135 140
Gly Gly Gly Gly Gly Gly Asp Gly Thr Thr Ala Ser Tyr Val Ile Ala
145 150 155 160
Glu Leu Leu Arg Pro Phe His Gly Ser Leu Pro Asp Ala Thr Leu Val
165 170 175
Met Trp Leu Ser Asn Ser Pro Ala Ser Thr Ser Gly Ser Asn Gly Gln
180 185 190
Trp Val Lys Glu Asp Val Arg Leu Pro Gly Glu Val Cys Thr Gly Thr
195 200 205
Asp Pro Phe Thr Thr Asp Leu Val Phe Ser Phe Gly Glu Ser Cys Leu
210 215 220
Cys Trp Ala Asp Leu Phe Met Gly Ile Leu Phe Cys Asp Leu Ala Thr
225 230 235 240
Leu Arg Ala Pro Arg Phe Arg Phe Ile Pro Leu Pro Lys Ala Cys Ser
245 250 255
Phe Asp Pro Val Gly Lys Tyr Gly Arg Pro His Met Pro Glu Phe Arg
260 265 270
Ser Met Gly Arg Val Asn Gly Val Ile Arg Leu Ile Asp Met Glu Gly
275 280 285
Phe Thr Asn Glu Tyr Leu Ala Val Asp Glu Val Lys Leu Thr Ile Trp
290 295 300
Thr Leu Ser Ala Asp Leu Ser Glu Trp Glu Lys Gly Pro Val Cys Thr
305 310 315 320
Val Gly Asp Ile Trp Ala Ser Glu Glu Phe Val Ala Met Gly Leu Pro
325 330 335
Gln Leu Arg Pro Met Cys Pro Val Leu Ser Met Val Asp Glu Asp Val
340 345 350
Val Cys Val Val Met Thr Glu Val Glu Ile Glu Glu Ser Asp Val Thr
355 360 365
Asp Phe Asp Asp Glu Gly Asn Lys Leu Lys Phe Lys Ala Gln Tyr Val
370 375 380
Leu Asp Ile Asp Val Arg Arg Lys Arg Val Leu Ser Ile Thr Gln His
385 390 395 400
His Ile Glu Ser Met Gly Asp Leu Ile Pro Asp Leu Ile Ala Cys Glu
405 410 415
Phe Thr Ala Tyr Ser Glu Leu Ser Lys Gly Met Gln Ala Met Val Glu
420 425 430
Gly Asn Glu Gly Glu Glu Ser Thr Lys Arg Met Lys Val Lys
435 440 445
<210> 5
<211> 443
<212> PRT
<213>Sc-ia albumen
<400> 5
Met Ala Pro Thr Trp Val Leu Leu Asp Arg Phe Val Lys Pro Thr Ile
1 5 10 15
Phe Asp Glu Glu Glu Ser Lys Gly Lys Gly Glu Ser Thr Gly Ala Pro
20 25 30
Val Lys Tyr Leu Pro Ala Arg Leu Arg Gln Glu Val Pro Ala Gly Met
35 40 45
Arg Asp Val Lys Pro Tyr Pro Glu Val Ala Asp Pro Pro Ile Val Ser
50 55 60
Arg Phe Ser Met Leu Ile Ser Arg Lys Ala Ile Arg Val Val Lys Ser
65 70 75 80
Val Arg Val Glu Cys Ala Asp Lys Ser Leu Val Leu Phe Tyr Ala Gly
85 90 95
Thr Gly Phe Pro Gly Phe Ser His Gly Cys His Leu Ile Tyr Asp Ala
100 105 110
Ile Asp Gly Ser Leu Thr Ala Val His Thr Phe Pro Phe Pro Val Ser
115 120 125
Gly Val Val Trp Val Gly Thr Ala Ala Val Leu Arg His Ala Gly Gly
130 135 140
Gly Gly Gly Gly Asp Gly Thr Thr Ala Tyr Val Ile Ala Glu Leu Leu
145 150 155 160
Arg Pro Phe His Gly Ser Leu Pro Asp Ala Thr Leu Val Met Trp Leu
165 170 175
Ser Asn Ser Pro Ala Ser Thr Ser Gly Ser Asn Gly Gln Trp Val Lys
180 185 190
Glu Asp Val Arg Leu Pro Gly Glu Val Cys Thr Gly Thr Asp Pro Phe
195 200 205
Thr Thr Asp Leu Val Phe Ser Phe Gly Glu Ser Cys Leu Cys Trp Ala
210 215 220
Asp Leu Phe Met Gly Ile Leu Phe Cys Asp Leu Ala Thr Leu Arg Ala
225 230 235 240
Pro Arg Phe Arg Phe Ile Pro Leu Pro Lys Ala Cys Ser Phe Asp Pro
245 250 255
Val Gly Lys Tyr Gly Arg Pro His Met Pro Glu Phe Arg Ser Met Gly
260 265 270
Arg Val Asn Gly Val Ile Arg Leu Ile Asp Met Glu Gly Phe Thr Asn
275 280 285
Glu Tyr Leu Ala Val Asp Glu Val Lys Leu Thr Ile Trp Thr Leu Ser
290 295 300
Ala Asp Leu Ser Glu Trp Glu Lys Gly Pro Val Cys Thr Val Gly Asp
305 310 315 320
Ile Trp Ala Ser Glu Glu Phe Val Ala Met Gly Leu Pro Gln Leu Arg
325 330 335
Pro Met Cys Pro Ile Leu Ser Met Val Asp Glu Asp Val Val Cys Val
340 345 350
Val Met Thr Glu Val Glu Ile Glu Glu Ser Asp Val Thr Asp Phe Asp
355 360 365
Asp Glu Gly Asn Lys Leu Lys Phe Lys Ala Gln Tyr Val Leu Asp Ile
370 375 380
Asp Val Arg Arg Lys Arg Val Leu Ser Ile Thr Gln His His Ile Glu
385 390 395 400
Ser Met Gly Asp Leu Ile Pro Asp Leu Ile Ala Cys Glu Phe Thr Ala
405 410 415
Tyr Ser Glu Leu Ser Lys Gly Met Gln Ala Met Val Glu Gly Asn Glu
420 425 430
Gly Glu Glu Ser Thr Lys Arg Met Lys Val Lys
435 440
<210> 6
<211> 443
<212> PRT
<213>Sc-ib1 or Sc-ib2 albumen
<400> 6
Met Ala Pro Thr Trp Val Leu Leu Asp Arg Phe Val Lys Pro Thr Ile
1 5 10 15
Phe Asp Glu Glu Glu Ser Lys Gly Lys Gly Glu Ser Thr Gly Ala Pro
20 25 30
Val Lys Tyr Leu Pro Ala Arg Leu Arg Gln Glu Val Pro Ala Gly Met
35 40 45
Arg Asp Val Lys Pro Tyr Pro Glu Val Ala Asp Pro Pro Ile Val Ser
50 55 60
Arg Phe Ser Met Leu Ile Ser Arg Lys Ala Ile Arg Val Val Glu Ser
65 70 75 80
Ile His Val Glu Cys Ala Asp Lys Ser Leu Val Leu Phe Tyr Ala Gly
85 90 95
Thr Gly Phe Pro Gly Phe Ser His Gly Cys His Leu Ile Tyr Asp Ala
100 105 110
Ile Asp Gly Ser Leu Thr Ala Val His Thr Phe Pro Phe Pro Val Ser
115 120 125
Gly Val Val Trp Val Gly Thr Ala Ala Val Leu Arg His Ala Gly Gly
130 135 140
Gly Gly Gly Gly Asp Gly Thr Thr Ala Tyr Val Ile Ala Glu Leu Leu
145 150 155 160
Arg Pro Phe His Gly Ser Leu Pro Asp Ala Thr Leu Val Met Trp Leu
165 170 175
Ser Asn Ser Pro Ala Ser Thr Ser Gly Ser Asn Gly Gln Trp Val Lys
180 185 190
Glu Asp Val Arg Leu Pro Gly Glu Val Cys Thr Gly Thr Asp Pro Phe
195 200 205
Thr Thr Asp Leu Val Phe Ser Phe Gly Glu Ser Cys Leu Cys Trp Ala
210 215 220
Asp Leu Phe Met Gly Ile Leu Phe Cys Asp Leu Ala Thr Leu Arg Ala
225 230 235 240
Pro Arg Phe Arg Phe Ile Pro Leu Pro Lys Ala Cys Ser Phe Asp Pro
245 250 255
Val Gly Lys Tyr Gly Arg Pro His Met Pro Glu Phe Arg Ser Met Gly
260 265 270
Arg Val Asn Gly Val Ile Arg Leu Ile Asp Met Glu Gly Phe Thr Asn
275 280 285
Glu Tyr Leu Ala Val Asp Glu Val Lys Leu Thr Ile Trp Thr Leu Ser
290 295 300
Ala Asp Leu Ser Glu Trp Glu Lys Gly Pro Val Cys Thr Val Gly Asp
305 310 315 320
Ile Trp Ala Ser Glu Glu Phe Val Ala Met Gly Leu Pro Gln Leu Arg
325 330 335
Pro Met Cys Pro Val Leu Ser Met Val Asp Glu Asp Val Val Cys Val
340 345 350
Val Met Thr Glu Val Glu Ile Glu Glu Ser Asp Val Thr Asp Phe Asp
355 360 365
Asp Glu Gly Asn Lys Leu Lys Phe Lys Ala Gln Tyr Val Leu Asp Ile
370 375 380
Asp Val Arg Arg Lys Arg Val Leu Ser Ile Thr Gln His His Ile Glu
385 390 395 400
Ser Met Gly Asp Leu Ile Pro Asp Leu Ile Ala Cys Glu Phe Thr Ala
405 410 415
Tyr Leu Glu Leu Ser Lys Gly Met Gln Ala Met Val Glu Gly Asn Glu
420 425 430
Gly Glu Glu Ser Thr Lys Arg Met Lys Val Lys
435 440

Claims (7)

1. rice hybrid pollen fertility gene seatScControl pollen fertility allele, it is characterised in that including in japonica rice Single copy alleleSc-j, its nucleotide sequence such as SEQ ID NO:Shown in 1；With the tandem-repeated copies in long-grained nonglutinous rice etc. Position geneSc-iaWithSc-ib,Sc-iaWithSc-ibNucleotide sequence such as SEQ ID NO:2 and SEQ ID NO:Shown in 3.
2. allele according to claim 1, it is characterised in that alleleSc-jThe protein sequence of coding such as SEQ ID NO:Shown in 4, alleleSc-iaThe protein sequence of coding such as SEQ ID NO:5, alleleSc-ibThe egg of coding Bai Xulie such as SEQ ID NO:Shown in 6.
3. a kind of method for cultivating the affine system of paddy rice hybrid fertility, it is characterised in that utilize genome editing technique, will containSc-iIn the rice varieties of alleleSc-iaOr/andSc-ibPortion gene copies rite-directed mutagenesis, reductionSc-iLocus Active gene copy number, hybridize affine system.
4. method according to claim 3, it is characterised in that for containingSc-ia、Sc-ib1WithSc-ib2Paddy rice product Plant, will using genome editing techniqueSc-ia、Sc-ib1OrSc-ib2In any one or any two gene copy carry out Rite-directed mutagenesis, reductionSc-iThe active gene copy number of locus, hybridize affine system,Sc-ib1WithSc-ib2Nucleotides Sequence is sameSc-ib,Sc-ib1WithSc-ib2It isSc-ibTwo gene copies name.
5. method according to claim 3, it is characterised in that for containingSc-ib1WithSc-ib2Rice varieties, profit Will with genome editing techniqueSc-ib1OrSc-ib2In any one gene copy carry out rite-directed mutagenesis, reductionSc-iGene The active gene copy number of seat, hybridize affine system,Sc-ib1WithSc-ib2Nucleotide sequence it is sameSc-ib,Sc-ib1WithSc-ib2It isSc-ibTwo gene copies name.
6. the method according to any one of claim 3 to 5, it is characterised in that be made by the steps and obtain：
S1. utilizeSc-ibThe G nucleotide variations that gene coding region is the 235th, design one is rightSc-ibThe CRISPR/ of gene specific Cas9 gene editing target spots, this special target spot sgRNA of construction expression CRISPR/Cas9 carriers；
S2. contained with the conversion of this genetic modification carrierSc-iThe rice varieties of copy allele are repeated, are obtainedSc-ibEquipotential base Because of specific mutations, andSc-iaAllele is the mutant of wild type.
7. the method for cultivating the affine system of paddy rice hybrid fertility described in any one of claim 3 to 6 is in paddy rice cross breeding breeding Using, it is characterised in that：Any one of claim 3 to 6 methods described is cultivated into the obtained affine system of paddy rice hybrid fertility and round-grained rice Rice varieties hybridize, and cultivateScSeat does not produce the hybrid of pollen abortion, for paddy rice cross breeding breeding.
CN201710476748.2A 2017-06-21 2017-06-21 Rice hybrid pollen fertility gene seat Sc allele Sc j and Sc i separation clone and the application in breeding Pending CN107164390A (en)

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CN113728911B (en) * 2021-03-12 2022-05-31 华南农业大学 Novel rice high-temperature reversible sterile line and two-line hybrid rice seed production system

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