CN114480724B - Dominant early allele Soc1 from japonica rice H5 And molecular markers and application thereof in cultivation of early maturing hybrid rice - Google Patents

Abstract

The invention discloses a dominant early allele Soc1 from japonica rice ^H5 And molecular markers and their use in breeding early maturing hybrid rice, said dominant early allele Soc1 ^H5 The nucleotide sequence of (2) is shown as SEQ ID NO. 1. The invention screens out a dominant early allele Soc1 of Huai rice No. 5 which is a main cultivated japonica rice variety from Jianghuai rice areas by constructing a recombinant inbred line population and combining genetic analysis ^H5 . The invention also discloses a dominant precocious allele Soc1 ^H5 Is used for molecular marker assisted screening by using the gene screening marker S10 of (2), and Soc1 is selected ^H5 By introducing parent and backcrossing and selfing, the hybrid rice combination which is early matured for 4-8 days and does not obviously reduce the yield can be cultivated.

Description

Dominant early allele Soc1 from japonica rice H5 And molecular markers and application thereof in cultivation of early maturing hybrid rice

Technical Field

The invention belongs to the field of rice breeding, and in particular relates to a dominant early allele Soc1 from japonica rice ^H5 And molecular markers and application thereof in cultivating early-maturing hybrid rice.

Background

In order to solve the contradiction of more people and less land, the rotation modes of double-cropping rice, rice-wheat, rice-rape and the like are generally utilized to improve the land utilization rate, so that the annual grain yield in unit area is increased. The hybrid rice has the advantages of high yield, wide adaptability and easy cultivation, the planting area accounts for 55 percent of the total area of the national rice, the yield accounts for more than 60 percent of the total yield of the rice, and great contribution is made to grain production. The cultivation of early maturing hybrid rice variety has important significance in improving rice production efficiency and increasing yield of grains all year round. But hybrid rice is composed of female parent (sterile line) and male parent (restorer line). Therefore, the development and utilization of genes, especially dominant genes, which can advance the growth period of rice without affecting yield are key to the cultivation of hybrid rice varieties with corresponding characteristics.

Researches show that the Ef-cd-OsSoc1 is a dominant early gene module which can advance the growth period of rice and has no obvious influence on yield, and the action mechanism is to regulate the growth period of the rice by utilizing allelic variation of a long-chain non-coding RNA gene Ef-cd in indica rice and regulating the expression intensity of the OsSoc 1. However, ef-cd encodes a long non-coding RNA (lncRNA), the variant type is single, and dominant Ef-cd alleles exist only in small numbers of early indica sterile lines. Therefore, the discovery of a new dominant Soc1 allele is of great value in improving the utilization of Ef-cd-OsSoc1 gene modules, and is helpful for cultivating diversified early maturing lines.

Disclosure of Invention

The invention aims to: in order to solve the above problems, the present invention aims to provide a dominant early allele Soc1 derived from japonica rice ^H5 And molecular markers and application thereof in cultivating early-maturing hybrid rice, wherein the genes and the molecular markers are used for auxiliary breeding, and can cultivate early-maturing hybrid rice varieties.

The technical scheme is as follows: the aim of the invention can be achieved by the following technical scheme:

dominant early allele Soc1 from japonica rice ^H5 Said Soc1 ^H5 The nucleotide sequence of the gene is shown as SEQ ID NO.1, the sequence of the gene is different from that of a late-maturing control, and the expression quantity of the gene is higher than that of the late-maturing control. The dominant early allele Soc1 ^H5 Huai rice No. 5, which is a main cultivated japonica rice variety from Jianghai rice areas.

The dominant early allele Soc1 from japonica rice ^H5 The molecular marker is located at Soc1 ^H5 The nucleotide sequence of the marker is shown as SEQ ID NO.2, a 31bp base sequence is inserted between 36bp and 37bp of the sequence shown as SEQ ID NO.2 in the late maturing variety, and the 31bp base sequence is shown as SEQ ID NO. 3.

The dominant early allele Soc1 from japonica rice ^H5 The primer comprises an upstream primer and a downstream primer, and the base sequences of the upstream primer and the downstream primer are respectively shown in SEQ ID NO:4 and SEQ ID NO: shown at 5.

The dominant early allele Soc1 from japonica rice ^H5 The molecular marker detection method comprises the following steps:

using SEQ ID NO:4 and SEQ ID NO:5, carrying out PCR amplification on the rice genome DNA to be detected by using the primer with the sequence shown in the step 5, and judging whether the molecular marker exists in a PCR amplification product.

If the size of the amplified band is consistent with that of Huai rice No. 5 (133 bp), the rice to be detected is homozygote carrying the premature allele, if the PCR product is consistent with Zhenhui 832 (164 bp), the rice to be detected does not carry the premature allele, and if the PCR product can amplify the parent band, the rice to be detected is heterozygote carrying the rice premature allele.

Preferably:

the PCR amplification system is as follows: 2.0. Mu.L of template DNA, 1.2. Mu.L of 10 XPCR buffer, 0.5. Mu.L of 1mM dNTP, 1.0. Mu.L of 1. Mu.M upstream and downstream primer, 1.0U of Taq enzyme, and ddH were added ₂ O makes up 10. Mu.L.

The PCR amplification conditions were: 1) Pre-denaturation at 95 ℃ for 5min; 2) Denaturation at 94℃for 30s, annealing at 55℃for 30s, extension at 72℃for 40s, 32 cycles of amplification; 3) Extending at 72 ℃ for 5min, and preserving heat at 10 ℃.

The dominant early allele Soc1 from japonica rice ^H5 The application in cultivating early maturing hybrid rice.

Said dominant early allele Soc1 derived from japonica rice ^H5 The application of the molecular marker in the cultivation of early maturing hybrid rice.

Preferably, the method comprises the step of introducing the dominant early allele Soc1 derived from japonica rice ^H5 Introducing indica rice receptor parent, and carrying out auxiliary screening by using the molecular marker to obtain the early maturing hybrid rice.

Further preferably, the parent is selected from indica restorer line zhenhui 832.

Further preferably, the dominant early allele Soc1 derived from japonica rice ^H5 After the receptor parent is introduced, backcrossing and selfing are carried out.

The beneficial effects are that: compared with the prior art, the invention has the following advantages:

(1) The dominant early allele of the invention is a new allele obtained by screening and excavating from a main pushed variety of japonica rice by the applicant, is different from the cloned Ef-cd gene, and is a new gene resource.

(2) As the early maturing genes are controlled by a pair of dominant genes, the early maturing hybrid rice combination can be obtained as long as one of parents carries the allele; the early maturing alleles of the invention also have the advantage that they have no significant yield loss, except for early maturation, compared to the control, and thus have significant breeding utility.

(3) The present invention is directed to the early maturing allele Soc1 ^H5 The intragenic molecular marker S10 is designed, so that whether a target individual carries the allele can be accurately judged, the breeding efficiency can be improved in hybridization and backcross breeding, and the breeding process is accelerated.

Drawings

Fig. 1: the flow chart of the invention is shown schematically.

Fig. 2: the heading date and heading date frequency distribution of the parent town recovery 832 (i.e. R832), huai rice No. 5 (i.e. H5) under natural low-temperature short-day (i.e. Naturalshortday, NSD) and high-temperature long-day (i.e. Naturalshortday, NLD) environmental conditions.

Fig. 3: schematic representation of a substitution fragment (genotype) of the introduced line IL 269.

Fig. 4: induction IL269 and BC ₄ F ₁ And (5) the heading period of the Huai rice No. 5 is compared with that of the Huai rice.

Fig. 5: BC (BC) type ₄ F ₂ Heading period profile of the population.

Fig. 6: the result of fine localization of genes is schematically shown.

Fig. 7: the early maturing of Zhenhui 832 was compared to the heading period in the control of high temperature long-day (positive period) and low temperature short-day (Hainan winter) conditions.

Detailed Description

The present invention is further illustrated below in conjunction with specific embodiments, it being understood that these embodiments are meant to be illustrative of the invention only and not limiting the scope of the invention, and that modifications of the invention, which are equivalent to those skilled in the art to which the invention pertains, will fall within the scope of the invention as defined in the claims appended hereto.

Examples

1. Identification of dominant premature QTL

Gene targeting populations: the late maturing medium indica rice restorer line Zhenhui 832 (plant new variety weight: CNA 20100442.0) bred by Zhensu hilly area Zhenjiang agricultural science research institute is used as a donor to be hybridized with japonica rice Huai rice No. 5, and then a set of recombined inbred lines containing 223 lines are obtained through continuous 8-generation single grain transmission.

Genotype of recombinant inbred line was detected using 102 pairs of SSR, indel molecular markers with polymorphisms between Zhenhui 832 and Huai rice 5 (table 1), one QTL detection located between chromosome 3 short arm markers In3-2 and In3-3 was detected under the short sun conditions In the hainan (LOD value 10.7281, 19.6293% of interpretable phenotypic variation), whereas the site effect was insignificant under long sun conditions (table 2).

For fine localization of the gene, a fragment carrying the 3 rd staining fragment of Huai rice No. 5 was selectedThe target strain takes In3-2 and In3-3 as foreground selection markers, carries out backcross of 3 generations and selfing of 3 generations, combines 102 to carry out background detection on Indel and SSR markers, obtains a substitution line (IL 269) based on a background of Huai rice No. 5 In 2017, and the marker detection finds that the IL269 is highly consistent with a recurrent parent background, and only 6 markers of 2 fragments of 3 rd and 4 th chromosomes are In contact with a donor parent (black section). The heading period is about 8d later than the recipient parent. In 2017, the Zhenjiang is backcrossed again with IL269 and Huai rice No. 5, and a secondary backcrossed population is constructed. Planting F in Zhenjiang in 2019 and 2020 respectively ₂ Population and derived F _2:3 Strains for fine localization and phenotypic verification.

The results show that: BC (BC) type ₄ F ₁ The population growth period is similar to that of the receptor parent, and is significantly earlier than IL269, but BC ₄ F ₂ The population growth period is separated, and 700 plants BC are planted ₄ F ₂ The group (G293) has 9 plants which can not be investigated for growth period due to factors such as plant diseases and insect pests, and other single plants are early matured: late maturation (bounded by 99 d) =525:166, consistent with a 3:1 split ratio, suggesting that IL269 carries a recessive late maturation major gene from zhenhui 832 (R832).

TABLE 1 102 pair Indel, SSR detection markers used in this study

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TABLE 2 screening of recombinant inbred lines of Zhenhui 832 and Huai rice No. 5 under different environmental conditions

2. Fine localization of OsSoc1

In order to locate the recessive slow-maturing genes, 15 extremely slow-maturing individuals are randomly selected as initial locating groups, molecular markers In3-2, in3-3 of sections where the 3 rd chromosome substitution segment is located and corresponding section markers In4-6, RM7187, in4-10 and In4-12 of the 4 th chromosome are utilized to find that the molecular markers are coseparated with the 3 rd chromosome markers In3-2 and In3-3, and the exchange rate of the molecular markers with the 4 th chromosome markers is close to 50%, and the genes are proved to be located between In3-2 and In3-3 of a 3 rd chromosome short arm again.

Then using two-wing markers In3-2 and In3-3 to detect all BC ₄ F ₂ The genotypes of the individuals were 672 individuals, no exchange occurred, and 19 individuals produced recombination between the two markers. Further, the markers were encrypted between In3-2 and In3-3, and the genotypes of these 19 recombinant individuals were analyzed, and F was combined _2：3 The postform represents the phenotype, which is finally located between Indel markers S10 and S26 (i.e. 1294602-1332821bp of chromosome 3 of rice), with a physical distance of 38.03kb.

According to a rice annotation plan database (https:// rapdb. Dna. Affrc. Go. Jp/viewer/gblow), the localization interval comprises 4 genes, namely Os03g0122600, os03g0123100, os03g0123200 and Os03g0123300, wherein the Os03g0122600 is a reported OsSoc1 gene for regulating and controlling the rice growth period. IL269 and parental Os03g0122500, os03g0122600 genomic DNA were sequenced and analysis showed that: no difference is found between parents of Os03g0122500, while a plurality of differences exist in the parents of Os03g0122600 gene, and further comparison shows that S10 is positioned inside the genes of Os03g 0122600.

TABLE 3 phenotype and genotype verification of key recombinant individuals in Fine positioning

Note that: b represents genotype consistent with Huai rice No. 5; h represents a heterozygous genotype; a represents a genotype consistent with zhenhui 832.

TABLE 4 part of the polymorphic markers of this study

3、Soc1 ^H5 Breeding utilization of (C)

From the results of genetic analysis and gene localization, it was found that the dominant early allele of Soc1 from Huai rice No. 5 (designated Soc1 ^H5 ) The heading period of Zhenhui 832 may be advanced. In order to develop an improved line having early maturing properties, allele Soc1 from japonica rice No. 5 was used as an intragenic marker S10 ^H5 The indica rice restorer line Zhenhui 832 is introduced, backcrossed for 3 times and selfed for 2 times, the Zhenhui 832 early line which is 4-6d earlier than the control and the hybrid rice combination thereof are obtained, and the yield test shows that the main yield characteristics of the improved line are all obviously changed (table 5).

Table 5, 2021, n Ji Zhenhui 832 and its introduced system mainly possess agronomic traits

The invention only lists one embodiment as a specific case of patent implementation, and obviously, all genetic modification and breeding utilization utilizing the gene resource or the corresponding screening marker can fall into the protection scope of the invention.

Sequence listing

<110> institute of agricultural science for Zhenjiang in Jiangsu hilly area

<120> a dominant early allele SocH5 derived from japonica rice and molecular markers thereof and use thereof in cultivation of early hybrid rice

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<170> SIPOSequenceListing 1.0

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agtcgccctt tgccgactct cagatcccct tgcgttttac cacttttgcc ttgtccgccg 180

cccctccaat tgggataaaa aagaaaagag ctgcatctct cgctgctcct cctcctcctc 240

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gtgcggggga agacgcagat gaagcggata gagaacccca cgagccgcca ggtcaccttc 360

tccaagcgcc gcaacggcct gctcaagaag gccttcgagc tctccgtcct ctgcgacgcc 420

gaggtcgcgc tcatcgtctt ctccccgcgc ggcaagctct acgaattcgc cagcgccagg 480

tacgcacccc ccctcccccc tcctcctcct gcccaatctg ctcttcttct tgtccccatc 540

ccccaatctt ctcttgtttt gttgcttagt ggtggggaac gggggtctca tattagttta 600

ttactagtct tttcctgaaa tccctaagaa gaaaacctgc gaggaccgat ggatcttctc 660

gctgcctgtg ccaaactcct gtagcaactg ccccccgccc ggcttgatgg cccttcgtga 720

tgcgtccgca gattttttct tgcaaatttt tttgtgggtg tgatgaatta ctagtataac 780

atctgcacta tgcagatgcc atgctgccct ctctctctcc tcccccaaga atctgcttat 840

cttgattatt tcttcgagct ctttgcctct ttcccccaac cgcacgtctc cgcccgagag 900

attctcgcct ggattttgcc gacgagtgcg tttctgatct ccttcctcca atcctcatgc 960

ttgcttgctg caatctgggc tcaactttaa agtgtccact gccgtctggc tataaggaca 1020

gttgctacta catgtagggg attcactatt atacccgata aatttaatga cgacggccaa 1080

taggaaaatt cggcctgaaa agacggcaaa aactatattt ccccgtgtgg cgattgaggt 1140

cagtctagaa atttgtcgag aacgaacgca cttaatgctt tgtctgaacg aacgcactta 1200

atgctttgtc tgaagaaata tttcagaatc cctagagcaa gtttaatagc agagctcact 1260

gcttattata agtcaatgtt ggagctaact tgtataacag gttagctata aggttaactt 1320

tattttttcc attctctctc tttctcactc tatgaattta atgcaaattt tttggagctt 1380

gtgtggagct ggctcttgta tgagagccaa cactacccac ttttttttat ctctctcttt 1440

tacataagca tatagcttgc ttatagccca ttactattct tgctcttact gtattttttg 1500

ggggtccggg ctgcttagac gtacggctag ttaatcagtg atttgttttg caatacatgt 1560

tctttcttgg gctacttgct gttgctatca tttgggctga catgacgtgc agaaacaaaa 1620

ccaatgcatg tgatagtgta ctgctgctgt atgcacgcac tgtggtggat tttatttcct 1680

ggccacgtac tgtattaact agatagaatt atcggttcca catctagggg tggtctattg 1740

cttcattaga gagtactact atgttgtatt cagtggtaca taaccaccgg ttctttacat 1800

ctcaccggct aatcttatca atttccaatt acacatgact taattagtat acgctagtta 1860

actagtgtat tagaaaagct atgctaagct actgaattgc caaacttcac tttgctttgt 1920

tcagttgccg tctaaacagt cacactactt ccataaagaa attagttgtg agactggcaa 1980

ggttagaaat tctgaaaaca taggcaactt agtactggca ttccggaggg caatatgata 2040

tgagcactaa ttagttaatg ataatgtacg ttgcagtaat cttgccgtgc atatgaattt 2100

ctatgcgtaa catgcatgaa agcacactaa cgggagtttc cagggagcct ccaatcttat 2160

taatgctgta caactaaaac aacatgtagc ttattgccaa aactgcatac atctaaaata 2220

ataatggata aaagaaaacc atgaaattct atgtctctga tctttctcgt cgtctgaggt 2280

cattacctga agataatatg taatgcaccc atgtgattga atacaaggta ataataatat 2340

atacatgaac atagggtggg cactgactaa tatgaattaa accaagataa tacttggggt 2400

ggaacctgag aaccacgtta ttatttagat atggcagccg accataataa tccagaatgg 2460

tagaatgaaa caaacatatt tgcttaccct acgattgtct attagtcaat atggatattt 2520

gccataatag acataaataa gttaatgtgg ttcattggtc ttcttgaata gttagtacaa 2580

ctagcatgta ttcaggtttg tgcagaaatt aaaacaggac acacatattt tatctgtcag 2640

ttattgtttt gagttaacct tactgaaata tatgtgtaaa attgtgtact gcaagattat 2700

tttcagctga ttttattgat gcatgttttg aattagaaaa ggcagcatta tctcctaaaa 2760

tgtgtcgtat aaataaatat gcattggatc tgttttcaac aaaaacatag actatggcaa 2820

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ggctcctctt accttgaatg caatagagct ctttgtcatt atggtttctt tgagaagaac 2940

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ttggagtaac atttcccatt acccctatgt acgcaatacc ctttcgtttg ttacatattc 3060

ccttaacttg cttacttatc ataatgccac ttgttagttt ccctttataa caattgtcat 3120

agaatagttc tttgtcgcat actccataga gcaacaaata acaattttag ttagtttgaa 3180

taaaaattct gtatttaaat ttcaagattt catacctata ggtgtatgta tcttttggca 3240

tgtttgcaac cttttatttt ggatatagac attttagttt ttagcatata catcaaagga 3300

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attacatttg gatcgctagc ttatatttct tttatcatgc atagcaaaga gaagcaaata 3420

aaattatttg gatgcaaagt ctgctaaatg acgatatgcg tgtgtgagtt cgtcattgtt 3480

tctacgtttt cttgctcttg tgggtgtagg cccaatttgg ttctctacca gcgcaaggtc 3540

tttgatgtca ttattagcct tttcttttct agcggagcga gaatcttcta gcattaccaa 3600

atggcatgcc tttcccatca aggataggat ttttcctttc tagatatcca agcgtaccct 3660

acattatctc acccaatgat acttctctat cattgattat tcttacttaa atttcaaagg 3720

attgtaataa actttcgttt tgtgcatgaa ccttcatact aaaagggtaa ataacacata 3780

gtttgttaat attaatttta tagataatat ataatgtttg gggctttgca taacatttgg 3840

gtataacatg atatcatctg ataaaagttg gtaaccaaaa gaatatatca gcatgcatgc 3900

tttcagcata aaaaaggaac gcatgagttt cgtaccaaat ttagtgaacc gttacatgta 3960

gagaaatgtg gtctcaaact caacttcaag atcataaaca gtgcacgtat cacttaaaaa 4020

aatgagagac aaaacattaa aaaaattatg aacacaagtt acaatatatt atcttgaaat 4080

accatctcat ttagaggatg agttccttaa ctcaatatcc aagtcttcat gatcatttgt 4140

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acatcacctt actttttggt acttataatc atacattgat gtgtcaagta tttagatact 4500

agtgatatgc acttttggat atattcattt aaactatatc cacattttca ccatgaatca 4560

tacaagattt agcataaggt tcattttggc tgaaaattat aacttaattc taaagttttt 4620

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gggtgatgca catcaattga tatctatgat agtcttatag tattagaaaa agtcatgttc 4740

atcctagaca tttgcatcat gctccaaagt atttttaggt gttcagtcaa agatcatgaa 4800

aagatagaac aaacacatat ggtggcttcc atggatgccc ctatgctact tttagtatta 4860

gaaggaaaaa tatttctggg gccttttgag gtgtagatga tttggtaatt ccaatattaa 4920

aaaaaaatta aatagttgta tccaagctaa ttcaacattc ataaataaga acatggaaaa 4980

actagagttg gggcaaatcc atcttgtacc tcattttgtg ccattgccat acaatattcc 5040

cgcggaaaac gtgtgtagtt tcatctagtt ttatttagat tgggagaacg aagtactaca 5100

aagacaacat caagtctgat gtcgatatat ctaagctagc gttgctactt tgacaaataa 5160

tgagctaggc actagtttga aactgccagg cactattgcc atatgttggt gattggtccc 5220

actagccttg tcacatgcaa agtccacgaa tcctcgtttt cgctccgtgc tacctctggt 5280

gaacgacaca tcccctaaaa gacgagcatt tatctctttc tagagaaacc aagaatagtg 5340

tatcgttgcg ttatccagaa cctatcgccc tattttatcc aaatgataat gttggctagt 5400

gccattgcac cattactcta agcctcctaa acacggagca agatggtcag taatccttta 5460

aaaaaatctc ggaccacata taaatgtaat agttatagct accactcttt gatgttcgaa 5520

aggaaaaatt caacacttta taaagttcaa tacctacttt acgtacataa aataattaac 5580

actatctttt gtatggtagg tatttcattc tttttcaatg aattgctttt ggatttctca 5640

tctaaccatt actctttaat ggccacacaa gaaacttgtc acaccctgcc ctttcaccac 5700

ctagcagtga cacctgagag ggtacgccgt aggaaatcaa cgcaagctga ctcccctgga 5760

accacgataa cggtactagt ctcaggatac aattgctggt accgcggtca atcgagatgc 5820

accatgtcaa ctaaacgcgg tacatatata ttataccttt aagagttcac actttcggag 5880

caacttctgg agtgtaatag gcaatcaggg tagcaacagc gaaaccttgt gtaggtagac 5940

ccaatccaca aacaacggtt tgggatttgt aaggaaaccc taaccatcaa gttcacttct 6000

cttctcgaga aggaaaaaac ggcggttgag tacatacgta ctcagtaagt catgggtaac 6060

tgagaatgca ggctggaaag gagggttggg tttgtttcgc aaattttgaa aacatttttc 6120

caaaacacta agtgtggcat tgtttaccaa gtggtcggaa agaagcatat gccttgatct 6180

tgtccgcgaa ttgctttgca acaactcaag cggacaaaac aaatattagt tggttaaaat 6240

taatctgccg gttgacctta tcatgacatt ggtggaacac catcagccct caggcagctt 6300

ccaactcctt atcatgacat tagacaaata ttagttcctt ttcactttca aaccatcaaa 6360

cgttagccca tctcatcaac tcatctcttt gggtcctaaa gccttccata gtctccaaca 6420

cacaatgctt tccttcggga aacctaatat atgacgctaa ttgacatcct gatctacaca 6480

agattgtttc agaccacgta atggtcctcc atcagtcacc acgactgatt ctggtaatga 6540

tttccaaacc cacaaaccac tttcgcaaag cataggcaaa caaatctacg tgagaggaat 6600

cgccacacat tcgcccatta ccatttgtaa gactcttcga attagtagac ctttacagag 6660

gttgtaaact tcacccacaa gatgcgatct tactccttca ctggccaagc tgggaaggtt 6720

tgtgacaagg ccttaccaaa tccaactagg ggtacctcat gtcacataga aggattctct 6780

ttgaactgag catcagggaa acttcggccc tcgtctacta aagaatacgg gatgccgcta 6840

ctcaatcgtg cataacacgt taaactttgt tttgtgccta gcgcaacaaa tcaactcaca 6900

tctcctcatc cggggcattc cccaactatt ggaaacaagg catacttggc caactattta 6960

cttagccaga ggtgccccat accacccata tagtcgcact atgagaagct cggataaaat 7020

taccaaggaa taagtcctta tatgcaaaat ataaaccacc gcgtgtcagt ttttaaaagg 7080

attcacaaca tatttttcca catcaacaaa caagggtgat gtgagctcat gagcacggtt 7140

taccgaggta ccacggggtg gagcgacaag ggattggggg tcatcagcgt cctattggag 7200

taatgcaact actgggtagg tctgttggtt cgtcgtgcga actgagacca gtcaaattta 7260

acgtgctcac tagcaataca attcgcaaac aaaacatttc taaaacaagg agcaatatga 7320

tgaaaaagtg ccttgccact gtcttcgtct tcgaagtctt aaaaaatctt cgtccgaaat 7380

aaattttgtc gtcgtcgact acgtgcatca aagagaaaaa taaacaaaat cctaaaataa 7440

aataccattg ttaataatta taagtgtcat ttggtagatt tcaatcttag aaaaaaaaat 7500

ggagacttga atgggatcga tcggagttac gaatagggat atatgatttt ttgaacattt 7560

aaatgatttt ctggaaatag aaaaggaatt atttgaaatt tggaaaagaa tattctttgg 7620

atcacaagta gggatgcaag cgggctgacc ggtgaaccta cttatctgtc aaataagtag 7680

taacccacgg gttcacgcat tgtggtccgg tttgtcgcac ctagagatac aagtggattg 7740

atccataaac ccatatatag atcaaattaa taggtaaccc gcgggtcaat ccgcgggtta 7800

tccactaatt ttgcttataa gcgggttcat gggttggccc gcttgcatcc ctaatcacaa 7860

gtgggtccac aagtcattga gtgggtgaag aaggagataa actggatttt tttaaggaaa 7920

ctgggggcac ggacttggtc tttagcccaa agaggaagaa gggtgacgag ggtcataagg 7980

acttggaaat gcagtgggtt gaagtcgacc tatgcacacg tgtcctaggt ggtgagaggc 8040

tggcgagtgg gagtgtaggg gtggtggccc gcagcggccg acgtggtagg ccatgcaggc 8100

tttgaccaag gaaggaggag actagctaga aatatcgatg acaggtggca ctcactagcg 8160

catcactaat ttacggtggt ttcaagccga cggagaccat tggcacattg agtgggcagg 8220

tgaagaaggg tgaatggctc ggatggggta gatgatgcat gataggggct agtagcgtgg 8280

agccaagtgt ggcggcgagc ggccctcgct gtgcggcgac caatagagcg gtgacttggg 8340

tgtgtgatgg ttcgggtgag gtgtgactct tcaccaattg aagatagagg ggagaaagag 8400

ggctcatcag aggagaagag gagaatggtg gcgacaaaac atcgacggat gagatttggg 8460

gaaaatagct ccatcgatga atagggctcg ggggatggct ggagcgtggt cgcttagtgg 8520

tgatgagtgt ggtggtggcg ttgagttggt tgttttgagg ttggtggtgg tggaaccgca 8580

gtgggcagag tgatagcgac tgacatggag cggcatcgtg aaaaggtaat ggtgtcggtt 8640

taggggctcg atatggaggg atggacgcgc gtaagacgtt gagcgggaag gatggaaaag 8700

aaaagaatcg agagaagaga gggaaggaaa gggatcataa aagagagtaa gatgggagga 8760

gagagtgggc ttgacaggtg gttctgtggg gagcacggtc gctaacatat ggacccgcgc 8820

gagaggtggt gcatgcatgc atgttgggcg tgtgccgacg gccttgcgtg agttaaggag 8880

aggggatcgg gccgttgctg atctgggttg aggggagaga gaggaggcta gtgttgggct 8940

aggcttgtgg tgcgcgggag acttgaggaa gagggaaggg gaggagggag aggaaatttt 9000

tagggaagct tttgaaaatt tctagggaag ctttctagac catgtgcata gtaacaagtg 9060

aaaatttcta gggaagcttt tgaaaggcta tttgaggatt tcgagttcgg atttgaatac 9120

aaatttgata tttgaactca acaaattaag gggattcgaa ggaaaggaac tcaaggaatt 9180

tgttagaatt tgataatgag aaattcaaga ggatttgaaa tacagaaagt aattttggat 9240

cggggttttt gtggtcgagg gagagattca attgagacct ttggcaccta tgtccttaat 9300

cacacacaat atttcactaa tgtggtaatg ggcaattgca tccattcccc cactttcaat 9360

cccaactgtt gttttacccc cactttttag ggtttgcaaa tttgccccca cttttttgaa 9420

aacaaacctg acggttgccc tcacttttga cggtgttagt tccaggggcg aaaggacatt 9480

tatatcttta gtggttgttg catttttgcc cccttttttt catcgtgtga ttttacccta 9540

ttatgtaatg atataagatt gtgataaata attcaaatat gagcacatct cgtgtgatgt 9600

ttttgtactt taacaaaaaa aatgacaaat actagtcaag tatagaattt gaacataatt 9660

ctagcagcat ttcaacagaa tttcgacaca aatagcacaa gtaacatata tatagcaaac 9720

attacaaatt gtccaacaca aataagccac aatacataga tgtcatagac cacaaacatt 9780

acttagtgtt acatagtcat aacaacacac attacatgtg cacaacatag aaggcacaat 9840

accacatatt acaagtccag atcatattgc aagtccatat tagcacaaaa gtgcacaaaa 9900

ttatttaaaa ggggcatata gatgtccaaa tcatgcaaac catgaagcta gcctccctcc 9960

tcttcattgc cttccccttc cacctgcatc gccactgcca cttccccttc cacctacatc 10020

gccactgtca cttacccttt cccttgcact tgcacttgat ggttaagaga gatcaacaat 10080

aacaaaataa gcataagcaa aatctgaaat aagcaaaaat tgaagataaa tataaggaaa 10140

gagcaataca tgactcactt tgatgtgcaa cacttggttc gttgttcccc tctctactgt 10200

tccaatttct aaagatgagt gcctcctctt ttgacttgct gtgggacctc cttggcaagt 10260

cttggccgtg tggcctagct cattacattc agtgcattat tttctcttac tagtgccagc 10320

ctcatcactt gcctttatcc tagccattct tggtctccca gggttccgta acaatttagg 10380

cttcttgatt ttatatccta tatcaacatg tccatctatc cttcaatgtc actggactga 10440

aagtccctac atatgccttc ttgaacttct caatggagaa ccagtcatcc acaaaatcat 10500

cgatatggac atccctacta agcttcgcaa tgaaagctaa agcatgaccg aaaggctttc 10560

cagtcacttg ccaaacccta caggaacatg tttgttactc tccaaatgta ctgcatgcct 10620

aaaggtgccc acagttactt cagctgttcc agcttcacat atcaataact catgttcctg 10680

gatattcttg cttttagcat tcagagactt tgtaatagct ggaattattc tttcctccat 10740

gttaccaaca atattccatc ttacaacaaa cttgtttgtg atgatctgtc ttattgcatc 10800

atgcatgtcc acaatccgtc gatccttggt tgttgagacc caactattga agcattcaga 10860

aagattatta ttgatgtaat ctaccttgca ctcttcagaa aatttggtta tgctctaaat 10920

gtatgaatga ttgttatcca accaggaaag tgcatcggaa cacttctcct ctatctcact 10980

catgaagctg ttggtagccg ctcacatatt ttgggcaaat agagggccat ggaacttctt 11040

tttcatgttc ttccacaagt gcctcatgca ttctctatgt tccactcctg gataaacatc 11100

atctacaaca ctctatattc ctttgcctgc atctatacta atgaccaaac ctacaatagg 11160

actatagttc ttggttattc agttgaaatt gaatagatgg agcatgcaag aaacaactta 11220

aagaatagac aacttttgga gtaccaatag ctttgttcat gttctgaaca aaccaagtcc 11280

aattttcctt agattcggtc tcaatcaccc catatgcact ggaaaaagtc agttatgacc 11340

atcaattgca acagctgatg ccaactggcc tctagacctt ccagtcaagt gagtggagta 11400

tatatctatg taacgcctac atcctttcaa aaacccattg atatagggtt tgagagcaac 11460

agagaacctc gtgaagcaga catcaccatt gcactcttta ctgtccaact catctaagct 11520

gcttggcact gatcttaata gctcttcacg ataggtaggc agaagatcgt agctgtcatc 11580

ccactttcca tttatcctct gaagacccta aattatagga tattaatatg gtgcttgcag 11640

atcctttgct ccccttgagg ggttatcccg caacaagtcc accactcgat caactaccca 11700

cttagttgag gccatcattt caccatactt gttgaaagat aaacaagtgt gctttggtcc 11760

acttgtctta acctacaaaa aaataatgaa tagctttcaa cttattgaaa gtaccttgca 11820

tccaatgtac ttgggggctg tagatgcaaa aaaaaggaca ctcgcaaccc tattccgctc 11880

ttttacaaat ggctctaaac atttttcttg tctttcttga tggtttcaaa agcatgatca 11940

ttcaagatag tatgatgggt aactgcgaac ttgcattgat cccaatcagg gaacaccaca 12000

tcaacatcaa tacaagcatc atgaacatga tatgcaaaga cattacttgt ctttcttgtt 12060

ggtttcaaaa gcatgatcat tcaagataat atgatgggta actgctaact tgcattgatc 12120

caatcaggga acaccacatc aacatcaata caaggatcat gaatatcata tgcatagata 12180

ggtaaatcgt cgtcgtcgtc atcctcgtca actatttctt gatcaggatc aaacttaggg 12240

tcagagcaat tagagtcagt agattcggcc aaatctgaat catagctact atcactatgt 12300

gccacaagtg attcagtgtc agagtatatg ccctcatcat caattcctac cgcatcgtta 12360

tcatttgaac cctttttttt tatttcttgg ctctcttttg tgtgtctctc atttgtggct 12420

ccctaatttg tagcaatctc acttatccat gctctaggcc ttactgaagg atgacaccta 12480

cattttgtgg gtgaaaattg caatggacca ttgaaattat tggcttgggc atcgacgcac 12540

acaaccccct tatctacatt gagtctaaac cactctagca tatgcttatg tgattttatc 12600

tcaacaatac catcatctaa agaacgccgc aaaatcaagt actgtttaga accccaaatg 12660

tagtgttcag ctataaaatt aacaacttgc aggagatcat agttatgcat atcaatattt 12720

tgggttggca aggttttaca ttggaggttt ttctttggcc ctccatccgg caaactaaaa 12780

aacgacttaa ctctaacaac taattccaag cgcagttttc agtgaaaaat tgaatggcac 12840

agccagcgac ttgattgaaa atctaagaaa gaaaggatga aaaggaaaga aattggtaga 12900

cgtgaataaa aaaattgaaa aaaatagtct tagcaatata ttgggaaatt gcgcggggaa 12960

aaagagtaat tgatgttttc atgatatatc atgccccgtt aaactttttc atgtaaaaac 13020

ttatagttga gaaaaaaatg tcaaagaaaa gagtacaacc tacagatcaa tactcaaaaa 13080

taaaaaatta gctactctat tttttcctga tctaaatgaa ggttttgaac tgagtggcca 13140

attcctggtt gtcagcagtg tattgtgtat gttctgcagc tagctcctat ccttttatgc 13200

ttagtagtat ttcttcgtcc tcgttgctca aacatgttct tcggtcctca ataatttgtc 13260

cgtagaggtt tgaaggccga tttggaggat atcgtagata caggtaccat caatacatct 13320

cgtgctagtt ttgaagagga tatgacatcc tgtgattatt ccacaacctg tgtacattga 13380

agtgcgacga ttcatgtctg atggcgatgc tatccaaata gttggacaac tctatcgtca 13440

acacttggtt gaactaccat tgccaccatg gctggaagat gagcctccac aaattctgct 13500

acactatgat ttttcttacc tgttgcaagg gcgggtgtgg gtatttgcaa tgtgactccc 13560

taaaacttta attcatactt ttggaacacc taaaatgatt tagtattaac atcagaataa 13620

tagttggaat aatcaacata tttagtttct ccaataagtg aacaaagatt gaggaatcga 13680

tgtatttccc tctaggatca agaataaatg caaaagcata caacatgggg aggtttttcc 13740

aatatttaag ataattttgt ttcacactaa agactgctgg tcctataagt tcatcactat 13800

catactcttt gaacaaagtg gcaatttcaa gaatcttatg caaatttaaa ttagctgttg 13860

gataatatac tttagaagtt cagataaaaa gaaattgaaa gaacctctca acaacatact 13920

aagtatatgt ctatcaatag cagttgccca tcacttgtca aaacatcacg tgtttgaaga 13980

aaaataataa gcaaatcctt ataatgtaat acactttaac attagataag tggcattcca 14040

ccgatgtttt gtactggtga taaacttacg tgcacgctca tccgatgcca cacagtacct 14100

tttccattgg tgcgatggtt agaacgtgat tgcttaacga atggcctcaa tgtgtacatt 14160

taataactta aacccttttt taacaattag attaataata tggcatgcaa agtgttgatg 14220

cagaagaatt tgtagatgca ttatccaatg tcgtgcccca tgtgtcgtgc agacactgct 14280

tgttgtgtcc ggccgtgaca atccgcaaag tagccgtgcc atgtcgtgca ggcccatggg 14340

ctatggcgtc ggtccatcca tggcatggtc ggacgggctg tgtgttgatg agaaagcatg 14400

gcggcctggg agatcaactt agcttcaatg caggtccaaa atatttataa gatgtgggtg 14460

tgccaactag ttttatcctg caactaacaa gatgtataaa ttgaagatca aagagccaat 14520

tggccaataa gccgatatac caatgctagc caatgtcgat accaacctat atctataggg 14580

ttttgatcaa tcagctatat gttcaatgta tattttgaca cttgagtaaa taatggtata 14640

tagacaatca actgataatg taattaaaca atgatataat ccaatagaaa ccaatcggtc 14700

aataatgata tgatagaata attgtttgat ccaaaggtta agcatacacc ggctggtgtt 14760

ctgatgtcag tgaaaccttt gtagcaatca gctaaaacca acttgtattt aatccttgta 14820

aacttatcct gaaaagaccg agttcaagga agaatcgtaa agaccacggc atgatagttt 14880

aattctattt attaattagg caagatttag taactttcct tttaacttta ggaagtattt 14940

atctagtgtt cgattgttaa catcaaaatt ttgtaagccc cgaagtcatc atcaatctag 15000

agtcagtatc ggcttcagag tcctgaaatc gaccgatgcg aattatctgg tggccaatag 15060

ttggattctt ggtagattcg gttaaggaaa ttaatcgatt aaaggaagtt tatctagtgg 15120

agaccgagtt caaggaggat gtggtatggc agtttatcta ttactccctc cgggctgata 15180

atacttgtcg ttttagacaa gggtaaggtc aaactttaga atctttgatt atgaatcatt 15240

tttttaggga agattatgaa ttatttttaa aatatttgtc tttcaaatat ggtgactaca 15300

tgtatagatt agtcttaaaa agtactttaa taaaatcata tatttgttaa cacttttata 15360

catattataa tagaaaataa tagttaaagt tgtttttttg gagatcgtgc ccttgtccaa 15420

aacgacaagt attatcaacc cggagggagt aattaggaga gtttgttagt tttttttttt 15480

atccttggta aaatgtgttt attgtttgat aagtattttt atgtttttct tttatcttta 15540

ggaaagtttc tttcttttcc aagaaggatt agtatctacc cttaggtata aatatgtaca 15600

cccgtagtcg ttgtaaacta tctcacagat caatataact actctttctc gaggttttat 15660

ttatcactgg cgaaacttgg cacctgacgc ggggctgtat cgattttcgg tcttcggcga 15720

aggggtaagt cctacgtttc gccggtcatg gcaattgtat cgactatatt ggtattgtcc 15780

aaggttgcat cgctgcgatc tcttggatcg ctctggttcg gatgatatat tttcctacct 15840

atttatcata tatctctgtt aatctagttt ggcagcgcaa tttccttgta tcggcttgag 15900

atctgcttca ggatttttgc cgatattggc taaattgctt tgtcagatta gattagctaa 15960

ggtatccacc gctcttaaaa tcagtcaaag gctcgattgt ttagacttta tgtttctttg 16020

catacttcta gttgcgtcga ccttagtcgt gcttagaacc ataaaaacta acctaccttt 16080

gattgtcaac aagggtttca tcagggtttc agctgatgag ttatctggac gttgtatcgg 16140

cttataagga ttacacacaa gttggattta gccaattgca acaaaggttt cactatttat 16200

ctaatcgttt ggattttatg atgtcggtcc ttcagccgat gtatgcttta gccttcggat 16260

caacatttgt tttatcatat cattattagc cgattggttt ctattagatt atattattat 16320

tatataacat tattatcagt ggattgtcta tatatcatta tttactcaag tatcagaata 16380

tacatgggat atatagccga ttgctcaaaa ccccatcgac atcgactggt ataagcatcg 16440

gctagtatcg gcatcagctg gaactattcc atcgacttcg atcggctatt tgatctgttg 16500

tttgcatatt ttgtcagctg caggatcaaa ctaactggca cgcccgcatc ttatcaattt 16560

ttggaccttc actagcgcta agcagatctc ccaggctgtt gctgttttca acttagttcg 16620

tgctaacata gatctggcac tatcggccga gcctgaattt ttcgtcaata gaatcttacc 16680

aaattttggt gttaacacgg tgctggcacg acctagtagc tcgtgggcct atgccgagtt 16740

agggcgggta ctgtagcaac gtgcctggct cagcccgata gagcatgacc cattttgcca 16800

gctttaggtg ggggtgagct tggggccctc cctctggaca aaggtcaaga cacaagctta 16860

gggtcttgcc ttgggtcatc ggatgggatc aaagctcagg atacggtcat ccctaccttg 16920

gggttagaat ccatggactg gttgtgacaa cagcaatgca ttaactactg aacgtgactt 16980

gttggtactg tcatttgcaa ggcaagcttt gcattaaatg ctaagtgacg catatctaca 17040

cggtgtgtga tgggagttac gtggtagcgg ttgtgcggct atcatgcgca ccatcatcac 17100

ccctacgaat agagcaaggc tttcagatgg catggttgac tgtcttgccg cttagacggt 17160

ctcagtgcat gtccacactt tcttccacct taggctgaca gtcggatcca gaaagcgtca 17220

atagtgctct aggtaggaca tgcggcaaag cgagaagttg tctttccaac gagcaccgag 17280

agggccacac aaggtcccga cgacctggtc cacttgtcaa caagcgacca tatttcatat 17340

acctaatccc tgtgcgctgc caggacctag ccgttagagt ttagccattg tggtattcct 17400

cgactaactg tatggcaagg tctaaggcca ccgggttagg ggatcgaacc ttagagatct 17460

ttcctcttag agatctttcc tctgttagtc tctgcatcga agagcatgag aagaacccca 17520

cgatattatg ctaagcgtgg cttgaacctg tataaccgac ttttcctcat atactcccgc 17580

aacaactcat gttctgcatt cccttgggtt gaactcacaa aggggcggtc aattgattcc 17640

ccgcttgaga gctcattccg agtatcattt tgggtcaata atgatgtcac catcacatgg 17700

ttagaggatg ttacaaacat atttatcaaa catgatacat ttgatatgta tataggagcc 17760

atcaacatca aatatacata ttaccgcaaa catatagtta ggggtgaaaa cggtatggaa 17820

atatcctgat cgcccgatac tgtttctgag aaactaatat aataataata ataataataa 17880

taataataat aataataata ataataataa taataataat aataataata ataataataa 17940

taataataat aattgtaccg actactacca tttctaaaac gaagtttttc caatttctac 18000

cagcgttaca ttgaacttga cactgaatag ttgaaaaaat atggaaattt cctgactgtt 18060

tttgtctgtt ttcgtaaaaa taataaaata atggtatcgt ttccaaccgt tttcccctac 18120

ccttaattac aacatcctat atacactttt cctataacca tcggttgatt ttcggaacat 18180

cgagaatctc aatatacgta actttatccc aatttctgtt gtaaaaacta ttccttgagc 18240

ccattatgta tatccaatct ttatatttga gaagatttgg ttgggatggt tgttcttgta 18300

aggaattgaa tatgtccaca ttaataaaga aaaacaaaag aagaattaaa aattaatatg 18360

atctagccat gaaactttag acattgtaac catattttta caaggccgac gtcgctgcaa 18420

cacatacacg accgatgtca tatattctgg acaatcgtgg gtgggaccaa aactttctga 18480

aagtactcat tgcacccggg taacctgttg aattagcggc taggtccatt tgacatgagg 18540

gttgaatccg tctccagaaa atgttacatt accttcgatg gcttgattgt tatgcggcgc 18600

catgagagaa gacttttttt tctgccgggg ggggggggag aggggggggg gccttcacat 18660

ggtcccgtat catcttcatt aataatccat tccttctatg ctcatgcttg tctttgcttg 18720

atccatgcag gtactaatgt taaaaagaaa tttgagatgt agtatctttg gttaaatata 18780

tagaacgcat gtgcctttag ttaattaata tgtaaacata ggcatggtca tatctaagac 18840

gattatgccc tcataagaca tcttaaatac gtagttcaaa tacatgaaaa tcctacacgt 18900

ggattgtctc gaaaggtatt ttataatagt attattttag taggttttat gtatatatat 18960

tgttcaatct tcaaaagaag agtttcacaa aacaacaact atttagtcaa aattagaatt 19020

ttgctacaac tttagacaga catttcacca aacaacatcg tagtggacga ttttactaga 19080

attttcacta ttatgagtgt cttgcatgtg gaccttggaa atagttagct aaagttgcat 19140

caaactgata attttgacaa aatggttatg atattctgag atgtactcaa aagaaactag 19200

tgtcaaagcc tagagtgaag agatcccaaa tcaaaaccct accacatgaa aattatagat 19260

tggtagtcct tagaaaggaa ttgaatgagg cacattgcat ttgagcatat ctttgtagtt 19320

gtgaaaaaat gcattaattg attgtggtca tttgcttgca aaatatattt cattctcttt 19380

ttgttgcttc tcttgtatat attttcttta atatatatat ttacagtggg ggtctccctc 19440

actgtttttg ggtcaatata tatatatata tatatatata tatatatata tatatattta 19500

tttatttatt tatttattta tttatttatt tcattctctt gtactagttg aaccccgact 19560

attatccttt tcgtgcatag tctttttccc atcaacgtat tgctaccgtt taattgttgc 19620

cccttttttt ttagaattgt gaccacaatt gtgagaacta tttaattcat gtaagaaaac 19680

tacttagcat gctaaatatg ttaacaattg tgaccacaat tgtaataaaa ctacttaatt 19740

catgttcttt tgcatgtctt gatgtagtct catctttatt gggcatggta attgaagata 19800

aattgcttga taacatattt atcgttttct atggcatttg tccttggtct ttttgatata 19860

ccaacaatag ttgttttatc atcataaaag aatacagtgt aattaagaat agtcattctt 19920

tttttaaaaa aaaaacatat tagtgctaag ataaatctag aattgaaaat ctttgttatt 19980

ttgtctccag tttggctgcc gttgtggtta tttgtggaca tatctttgta aaatgggtgt 20040

attttggcaa tggtactttt agatgcttta attatactat tatgtctatc ttctctattt 20100

tctcatatgg agtgcactta ttttgacagc atcactactt agctcaaaat ggagatcttg 20160

ggtcaatgag agaacatgct cgatacacaa aatagctacc acatttttgc tattttattg 20220

atatgtagca tgtgatgttt gtcttaagca cttagcaaaa aaaaaatgta actatcgatg 20280

taaattgttt tttttttctc tataaaatat ttgcctcatt atttttagtt gttatgttaa 20340

ttacacagta tttgcattaa aaaagttaaa attcgctatt gtggtgcatg cttattaatt 20400

taaaattata aaatttgctt tatttccata gttatttgag ttagcttcac ccatatgagg 20460

ccataccctc ccaaagacag taaagttgta ctcttgtttt tcatagacct tggctaggtt 20520

gagttattta tttgtacttg aacctatgca gtcaacctcc tttgagattc atattagatt 20580

atatatggta cctagtatta gatagaaaat atgacaatta ttatatataa ttaactgcca 20640

tttaggaaga ggaaaggtgc acattagctt ccaagatcta tctagcccat ttgtaggtgc 20700

attgtctttc aaccaaagtc cggctattgt ctgactttgt gaatatgaat taacccacaa 20760

ttggctataa tgcaagagtt tttatgcttg gctcaatacc aacacaccat acccttgcta 20820

gaaccttact ttatgtcgta tattgctcaa ggataactaa ttattgatca aaaaaattat 20880

gttatttaat acccataaac caagagatgt gcaaaaaaaa tattctaatg gctattactc 20940

caacactaat gtgatcaagg ataaaataga aatcaaggta aaaaaaacaa atcctttagt 21000

ttcttacttc agatatgtta cacccaaaag catatatatc atcaaggtaa attggttagt 21060

atagatcgtt gctaatttat ggtaaaatca tttgagcgat tctgagtaag gaaaattgag 21120

aatttgcata acataatatg tgcataggtt gattaaagaa tttaatatta gaatgaaaga 21180

tgtgttagca ttgatgcata agaaatagtt tgaaattgtt gtttttctag taatatgtta 21240

tgttcttttg catgtaattt gaatctcaaa agataatctt ctcatctttt ctaatccaaa 21300

ctacactagt cccaaagtcc gacaaatttt tgtgttgatc tttcctttct tatctcttcc 21360

cccaaaatca tgcatgttaa atattgtctc ccttcaccac tatctacaac tctactctcg 21420

cgcgtttcat ggtctttctg tgttactagt tgcctggtgc aactcccatc atagccaggt 21480

atagatggcc atgtgggccg cccggcacgg cacggcccaa gcccggccgt gcctgggccg 21540

aggcttgtcg ggccggcacg gcccgaccga cgcaccgggc cgtgccgtgc cagcccacgt 21600

gccgcactca cagcccaggc acggcccaac acgcctcggg tcgtgccggg ccggcccgaa 21660

ggcacgatgg gccatcgtgc ttttcctcag aataagtcta aattttctcc ctcctattgg 21720

gctgtgacat tgtatagcta aaaaaagtct attttacctc cctcttcttt gtgatgtggt 21780

atataaatag ctaaaataag tctatttaag gtccctattc tttagacatt gacatttata 21840

tgtctatttt tagtccctat actttgtgta ccgggtctgg cctgccagcc catcgtgccg 21900

ggccggccca accgtgccgg gccagcccag cgtgctggtg gggaggccca ggcacggccc 21960

gacggtcggg ccgggccggc acgggcccga cggtcgggcc gggccggcat gggcccgacc 22020

caagtcgggc cgtgccgtgc ttgggccggg ccaaaaaggc gtgccgtggg ccgggccttc 22080

aggcctcggg ccttatggcc aactatatag ccaggttatg cttagattca tcattgatgt 22140

taattcctac ttagttgaaa tatatatttc tcatactaat cccttgttat agttctctac 22200

tgaaggttat aaaacttggt tgctaaatca cctatcatca aaattatcat gtaactagct 22260

attctgttta aaatataggt ttatatacat atatcttata tcattcagac aaaaacaagc 22320

aaatgtattt ctttttatgt atataggctt aaacagtaga acaatcaacc tagagaaaaa 22380

ttaagttctg atttcgtcat ataattgtaa tttgtatgta aaaaaaacat acttataaat 22440

tggtgaaaga gaaggattca aacacacaaa aaaaaaacta attcatcttt accatgtacc 22500

tataaattgg aaatctagat gtaaaattag tatatattgt tccaacttga tcacatcaaa 22560

gttccaacca taccttaatt cattatacaa catcaatcag tgcatgatgc ggatttctta 22620

aaacataaga tttgtggata tgaaaaaacc catatatcca taactactaa gtttctatgc 22680

aaggaatcaa taatagacgt catacgctat ggtgtaattg tggctttgag ccttggacca 22740

caatgaatga cggaagttta gttgaactaa ccttgtgtga tcaccatgta aattggtagc 22800

tagctaatta tctactcggt ttggtatatt tgtgacagga tttggatcct ataacgagaa 22860

gttagcttca tagacctggt tttagaatat tcccccaccg ttcttttgat ctcacataga 22920

taaaggggga aggacaaaaa cacaacatgg tcaatgcatg gtttaatggg ttacgatgca 22980

ttttacattg attcataaaa agccaagtat ttataatgtt tgtatcaaca aaagacattt 23040

ataacatgga agtggtaagc tcataccatt aggaagttta actatttgtg ctcttataac 23100

ccgtcgtcct gtgtttgccg ttggagagaa gatttctaaa tactatgcca ctatgtgtaa 23160

cacattggca tcatatggat tgttgatgtg agatcccaat tttgccccga tcaaaaggaa 23220

gaaacgatta aggatgtcca tggctgtcct tggccttaga tctgcatttg catgtagcgg 23280

agctgaccac cgtggccgtt gatgtagtcc tcgtcgacaa actccaccct catgaaatta 23340

gatgcttgta gttggatgta gaggagctca ccactatgct gttgatgtaa gtccttgtcg 23400

atgaactcta tctcgggtga gccaccatgt tatccatttg cttattggtc gctatgcacc 23460

tcaccatcca cctatgccac ctcgcaaccc ccgtaacatc gggcaagcac gcgcggcctc 23520

ctccttcctt ttgatgcaac accccacaac cactgcccac tatcatagcc cctcgccaca 23580

cggcaacaaa tcgaattaca aaagatagtc gtgcctaaga actcaatggc tagcaacagt 23640

atcctcccac gtgaaaccat gctacgagtt gtggaagcaa aggaggcgaa agaagaggaa 23700

gagaaggagg gttgttgcct cccgccgcca caactagtct actcttcttc aaggccctct 23760

caccatctgt tgctgacgtc cctggcaagg acgatgatcc ccatcgacct tcttggtgga 23820

agctttcgcc acatgccctt ctctatccaa ttgaccctcc aagtccacca tcggccccaa 23880

catcatcaat ggttctactg agctcccgcc accttcactg tcgtcgcaaa gaaccatggc 23940

tcacataggg tccccagcga cagctccata gcatctggtg gttgcggcag tgacttccac 24000

gggtaccaga ccttggggga aggggctgga gcaaaatgaa cttttaatgt caatcttcca 24060

tgttggaata cccatagtgg gaaaagtggc acaaggttaa gaactattct cattagaagc 24120

aaacatatag gaacaattgc aaaagtggca agtaattaaa tttttccata ccacttggaa 24180

tgcttagtga taaatgtttt gtttcaatgt tttccaagat gcaataaatg tcttcacaag 24240

ctaacaaccc cacactacac gactttgccg ctactacaat accattgtac aagtcataaa 24300

ataatagcct ttaccactcg tatgtatatc tatttttcac ccttgaccca agaaggatga 24360

attgaactca cagttgatat acatgtgcat cagttgaacc attgtgagag tagacccacc 24420

aatgtaagat ataaacccaa gtacgtgtca aatatgttgt agttctttgg ttggcccttt 24480

gactcaaagt gagaaaaagg tgtaatttga cttagcatgc ttcttgtgta tattcaaaat 24540

gtaatttgca ataagcctat tttagtggtc gaaaaatgtg tatacattag actcaagtca 24600

aatataccat ttatctcgtt aagatgatgc ttcgagacga gaataaatac tatatcataa 24660

tgacaatggg tatataaatg gtggttatcc tacaattaaa gtccaaattt agcccttcaa 24720

ctattgcgtt gatctaattt ttgtcctttt aactataaag caattaatta ctgatcccgc 24780

aactttaaaa atggttcatc aacatggttc cattgatgtg gtgagatgga tccaacatgg 24840

caacggggtt gggtcggttc gtgttggaca aaaactccca tccctgttcg cttgcaggtt 24900

cacccggtcc cgaccccgaa agaaagaaca ggttgaaatt agtccctggc cgaaaccccg 24960

gcccctatag ggcctcgcac ttcagccaag tcccaacatc ccctaagcaa gcggaagttg 25020

ccgacggttg gaaggaggga agaacgacaa gtgccacctt caaccaaata aagcaatcgt 25080

atggagagaa gagacaacgt cgggaggcga ccgaagaggg agaggaatgg agtgggcgcc 25140

ggtggccgac cacaagatga ggaagaaggt gtggggcacc tgtggacaag atggtgtggg 25200

actccggtga tagcggcggg cgggataaag tcgacgcatg ggcgacgtgc gtgactcgtt 25260

gcatgcgcag agcattgaag cgatcgcaca aatcatgcag ccgatgggag catcgaattg 25320

tgcaggtgga ttattgtttt tttgtccttt tatattctat ttttggcctt atgggctttt 25380

gatggctaga aatacataca tgaggtgact atttcttaaa cttcagggct ccacgagtga 25440

agatgtcacc tcggccctaa caccgctcaa agtcagggcc ctactcagta acacccacgg 25500

gtgaagattt ggccctgtcc ccggccctgt aggggtgggg acccaccgag gccctgtttc 25560

caatggggaa aatgccaccc ctacctactt gtgaaaagag ttgttcacca cacaaatcaa 25620

caataaacaa caaagaaatg tagaaaatat tgggacccac ttgtcatgct caccaccttt 25680

ctctcttatg tatcttattt attttagttt gatagctact acttaattct atgttatcag 25740

tcattttagt tttgtcctat gttaaacttc ttttaacttt gaccaaattc ataaaaaatg 25800

taacaacttc tataacatca aatcacattc attaaatcca ccatgaaata ttatggatgt 25860

tggtgcatcc ttctataaac ctggttaaaa tttgattttg gacaaactaa aatggcttac 25920

aatatggaaa ggatggagta gatgctttgt ggcatccaaa tcataccacc aattacttca 25980

ccgtgtcata aaaaaaacct ggtttgggct tagggatagc tttattacga ggtacaaaac 26040

tgagcaattt tatagtcgaa ggtcaaaatt cagatcaagg ccatagttga gtggttgaat 26100

atggacgttt tccttgacct tctcatagag gatggataca tgggagaaaa ttcctacaac 26160

ttatttatgt ttaccaatga ttctaacctc caagagtgac ttgcctctac aaagcttttc 26220

tgaaatgacg tttttaccat gttttctaga ttctttatat acttaagcca catcattttt 26280

ggtaaaatca accatttcca tctgaagaaa ttttcataag acgttcatca atacacacga 26340

gccctttgga aaacaactta taagtgaagc tacttagaca aatagcaagt tcgatacact 26400

agaattgtga gaagccttct ttggcttcat aaagaatttt agtagcggga ggctgccatc 26460

atgggtcaca gttgatatac tataatttac tctatattta tttctttact cttactaaag 26520

aagttgttag gtgttggtgt attctacttc catccagcca aaggatttca tagattattt 26580

attttaaatg ggtcccgaca agtgcatatg catgtggcta attgatctta tgttaccgtg 26640

catcaggttt tagatgaatg gagaatttct ttctgcaatt tattttgttg ataaagtata 26700

ctccctccgg attgataata cttattgttt tggacaagag cacggtctcc aaaaagtaac 26760

tttgacaact attttctatt ataatatata tagaaatatc agcaaacata tgattttatt 26820

gaagtacttt ttaagactaa tctatacata tgatttccat atttttaaga taaatatttt 26880

ataatttatt tatagtaaaa gattttaaag tttgatttta cccttattca aaacaactaa 26940

tattattagt ccggagggag tattaattag cacatcttat ggcgaagttg acttgttatt 27000

ttaataactc tttatattgg catcgcctgg gaagaatacg gtgccacaga aaagcacccc 27060

agttttaact ggttacatga ctttaacagc agcagcaaat acacttagtt aaccataact 27120

ccttttcatt ttcaagactc cattaccaag ttctaaaaag gcaactgttg aaaatgtctg 27180

atgattagtt caaatcaaat aaacgcaagt ttctgtcttg aattttgtat catatgtctg 27240

ttaattttag ttattatttc atggttttcc tcaaatttta tcaaaatgtt ctcctaggtt 27300

tatcaaaaaa atgagcaaca atgtgatatt gttttgttca gcagctgtat atatgattat 27360

ttcattcgca cgattccatt tgtaccagtt accacattaa tttgattatt aggctgtgga 27420

taataaagac cattcaatga taagcttggg caaagtttga gaccgcagaa tgaaactact 27480

agaagttctg gaacagttat accttttata aattcttctg gtgtttttag cagataagcc 27540

ctaaatttta ttctgcattt aagtcaggaa gatgttggag gactaccttt atctgtagct 27600

agtatatcta tttttttttc ttttgaacaa tatctaccta tctatatatg tctgtagttt 27660

atcatccggc aatttatatt ggatataggt cacacagtga ctcgaaaagt atgcaacaat 27720

ccaacaaaca ctcctatgtt ttcttgatag tactagtgta aatgcccttc caacttcatg 27780

agttttcttg atacatatgt ggggtgttct ggcttaagat atcaatatga cagagatcaa 27840

acatgtatag ttagattgat cttttcatga tctaaaagat tgatcttttc tccttatgta 27900

gcatcagaag atcgaaaggc gaaaaacctc tctagttgca gaaagaagtc aacctttgac 27960

cgcgttcttt ttcttaatta acaatttatg aagctaattg gatatttaat tgagatatat 28020

ctaacgtttt gtgcttttgt aattgaaaca gaattctaac actattctga cataatcttg 28080

ctgcttttct ctttacttca gtacgcagaa aacaattgaa cgctatagga cgtatacaaa 28140

ggaaaatatc ggcaacaaga cagtacagca agatatagag gtatataatt cttgttgaca 28200

aacttggtca tccttgtacc atagtgttat tgctgataga ctgattttag tataatttat 28260

gtcatatgtg tgcgtataat ttccgttggc agtgtcaatg ctgctccatc aatagatttc 28320

gttgtatatt gaggtaaaac gaaatgtata ttaagttagt tattttgggg aactctgttt 28380

tcaaatttta tttgaagatg ccctatatct gatagattgt atttttttta ataaaaaaac 28440

agtagattat ggttatgtat aaataagcat atataagatc cctgcactga agcagggaca 28500

gtactatttt taagacaaag aaaatcagat gtattctaga cttttaagaa tcagttttca 28560

attcaccaag gatgtgcata gtcttcgaga ttatggtagg aatgttttgg ttgagaaata 28620

aataaagaac atgagcacta aacaagcaat gaaaaaatgt gttctgaaat aaaatgtgct 28680

tctagtcgtt ttattgtagc tggacaattt tcaatagatt gtttggttgt ctgaattgta 28740

gcaagtaaaa gctgacgctg atggtttggc aaagaaactt gaagctcttg aaacttacaa 28800

aaggtacccc cttcagctta attacagaag tgccgtcctt gtagctgtca aaaaataggc 28860

tgttatcatc gtgtttaatt tgttgatatg cagaaaactg ctgggtgaaa agttggatga 28920

atgttctatt gaagaactgc atagcctgga ggtcaagctg gagagaagcc tcattagcat 28980

caggggaagg aaggttggct aataaatttt ccatttcagt ctacgttatc agcaattaat 29040

ttatctatat atatgatgtg cacaaactta caaccaattc tctgcatact agacaaagct 29100

gcttgaggag caggttgcca aactgagaga gaaggtacct caccaacttc gtacttacta 29160

taaataaata aatatatata taacctgaaa cgcaggacca atcatctgtt cctaatgtcg 29220

tgtttgtctc ttttttgcat taacatcgac caggagatga agctgcgcaa ggacaatgaa 29280

gagttacgcg aaaaggtaag acgagtgaaa cttttactgc cgagaagaca ccagcagcca 29340

gcttaagtcg atcgcccagc ctagcatatg attatttgag ctaattcgtg cgatatctta 29400

tgtgcacacc aatataaatg ccgagaattt tctgcgacac tgaaagatca actcacctga 29460

tttgcttccc ttgatcatga actttgtgca gtgtaagaat cagcctccct tgtctgctcc 29520

tttgactgtc cgggccgaag atgagaaccc ggaccgtaac atcaacacca ccaacgacaa 29580

catggatgtc gaaactgagc tattcatagg gctgcctggc agaagtcgct ccagcggcgg 29640

tgctgcagaa gatagccaag cgatgcccca ttcttaagta acaggccagg aataagctgg 29700

atctctgcgt tgagaaagga gatgatgcta ggcatgcaat gacaccaaac catcctaaaa 29760

acagaccacg ctgtatattt tacttacccg tattgtcgtc catggatgcg aaatgctaga 29820

cgcatatgca cccacccatt cagcaccacc aggcagaaac ccttcgatcg gcagactcca 29880

ctacccatct aaatctaaat cgagagatct caagtgcata cttgt 29925

<210> 2

<211> 133

<212> DNA

<213> Rice (Oryza sativa)

<400> 2

gatgttcgaa aggaaaaatt caacacttta taaagttcaa tacctacttt acgtacataa 60

aataattaac actatctttt gtatggtagg tatttcattc tttttcaatg aattgctttt 120

ggatttctca tct 133

<210> 3

<211> 31

<212> DNA

<213> Rice (Oryza sativa)

<400> 3

tcaataccta ctttacgtat aaagttcaat a 31

<210> 4

<211> 20

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 4

agatgagaaa tccaaaagca 20

<210> 5

<211> 20

<212> DNA

<213> Artificial sequence (Artificial Sequence)

<400> 5

tgatgttcga aaggaaaaat 20

Claims (3)

1. Dominant early allele Soc1 from japonica rice ^H5 Is characterized in that said Soc1 ^H5 The nucleotide sequence of (2) is shown as SEQ ID NO.1, and the molecular marker is positioned in Soc1 ^H5 The nucleotide sequence of the marker is shown as SEQ ID NO.2, a 31bp base sequence is inserted between 36bp and 37bp of the sequence shown as SEQ ID NO.2 in the late maturing variety, and the 31bp base sequence is shown as SEQ ID NO. 3.

2. The dominant early allele Soc1 derived from japonica rice as claimed in claim 1 ^H5 The molecular marker detection method is characterized by comprising the following steps:

using SEQ ID NO:4 and SEQ ID NO:5, carrying out PCR amplification on the rice genome DNA to be detected by using the primer with the sequence shown in the step 5, and judging whether the molecular marker exists in a PCR amplification product.

3. The dominant early allele Soc1 derived from japonica rice as claimed in claim 1 ^H5 The molecular marker of (2) is applied to auxiliary screening of early maturing hybrid rice.

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