CN105861518B - One rice seedling yellow leaf gene and its detection method and application - Google Patents
One rice seedling yellow leaf gene and its detection method and application Download PDFInfo
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Abstract
The invention discloses a rice seedling yellow leaf gene, which has nucleotide sequence shown in SEQ ID No.1, and the protein of gene coding has amino acid sequence shown in SEQ ID No.2.By carrying out PCR amplification to the extracted DNA of seedling stage yellow leaf rice, sequencing is carried out later and compared with common rice such as " Jiahua-1 and OryzasativaLcv.Nipponbare ", it was found that there are two the missings of nucleotide (TG) on the tenth exon for the gene, subsequent cryptographic is caused to change, coding amino acid number and type change such as SEQ ID No.2, so that there is seedling stage yellow leaf phenotype.The specific dCAPS PCR primer such as SEQ ID No.3 and SEQ ID No.4 is devised to this, by passing through after carrying out PCR amplification to DNA extracted in riceEam1105I digestion is identified, the rice plant containing the gene can be quickly detected, and detection accuracy is high, easy to operate, low in cost.
Description
Technical field
The present invention relates to Chlorophyll synthesis, Development of Chloroplasts and the photosynthetic machines of Plant Biotechnology and agricultural such as rice
System, the fields such as gene function, plant physiology, plant genetics and breeding especially on molecular level.
Background technique
Rice is second largest cereal crops in the world, and one of most important cereal crops of China.Therefore how to improve
The yield of rice just seems solving increasingly serious food problem also more important.Photosynthesis is to influence rice yield
One of key factor, and blade is to carry out photosynthetic vitals, leaf color largely determines photosynthetic
Efficiency.Chloroplaset is that higher plant carries out photosynthetic place, it is semiautonomous organelle, from proplastid development,
Its normal development needs the mutually coordinated of karyogene and chloroplast gene.Natural or non-natural factor can all cause to close with chlorophyll
At or the relevant gene of Development of Chloroplasts mutate, cause Chlorophyll synthesis or degradation to be obstructed, Development of Chloroplasts defect, finally
Cause the variation of leaf color.Research shows that the gene of control Development of Chloroplasts is destroyed and can lead to Development of Chloroplasts and be obstructed, and then make
Plant leaf color is abnormal, or even declines photosynthetic efficiency sharply.Currently, research workers utilize physics, chemistry, life
The mutagenesis such as object obtain rice leaf color mutant, be applied to the genetic breeding of rice, the clone of physiology and its related gene and
Functional study.This research and utilization physical mutagenesis obtains a rice seedling xantha mutant and is navigated to by map-based cloning
The gene of one control rice seedling yellow leaf, there has been no the document reports with rice this gene-correlation so far.
Summary of the invention
The purpose of the present invention is to provide a rice seedling yellow leaf gene, the protein and this gene of gene coding
Detection method and application.
If the allele is destroyed or can lead to rice seedling yellow leaf by the yellow leaf gene substitution in common rice
Character.
This rice seedling yellow leaf gene has nucleotide sequence shown in SEQ ID No.1.
The encoded protein of the gene order has amino acid sequence shown in SEQ ID No.2.
A pair is for detecting the specific dCAPS PCR primer of said gene, sequence are as follows:
Upstream: 5 '-GTGTGTACTCTCAGATGAAGTACTTGGACTCTCAGT-3 '
Downstream: 5 '-CACATTGTATTTGCAGAACA-3 '
That is sequence shown in SEQ ID No.3 and SEQ ID No.4.
The method for detecting this rice seedling yellow leaf gene is, using above-mentioned specificity dCAPS PCR primer to water
The rice seedling phase, extracted DNA carried out pcr amplification reaction, obtained 113bpDNA sequence as shown in SEQ ID No.6, will be obtained
113bp segment (Fig. 2) is usedEamIt is still 113bp(Fig. 3 after 1105I digestion), illustrate to contain the rice seedling yellow leaf gene.
In present invention research, the specific dCAPS PCR primer and detection method of inventor's design can be to rice seedlings
Extracted DNA carries out PCR amplification, the restriction enzyme site be using dCAPS Finder 2.0 (http: //
Helix.wustl.edu/dcaps/dcaps.html it) designs, recycles software Primer 5.0 to design specificity later
PCR primer, product are usedEam1105I carries out digestion identification, can be quickly detected the rice containing this gene according to digestion banding pattern
The accuracy of kind, detection is high, easy to operate, low in cost.If containing this yellow leaf gene in a rice varieties, with
The specific fragment (Fig. 2) that length is 113bp can be amplified by stating primer, then be usedEam1105I carries out digestion to the segment,
Its endonuclease bamhi size is still 113bp(Fig. 3).
Material containing a rice seedling yellow leaf gene is passed through by japonica rice " Jiahua-1 "60Co gamma-ray irradiation, warp
Hainan, Shanghai two places are for many years from accompany each other generation and selection, it has also become the stable strain of various economical characters.Inventor under study for action will
This strain and long-grained nonglutinous rice " Peiai 64S " prepare Its Reciprocal Hybrids, construct genetic group, will be contained using SSR and InDel molecular labeling
The one rice seedling yellow leaf assignment of genes gene mapping is on the rice chromosome of rice.
The present invention utilizes RNA perturbation technique, and the cDNA and pTCK303 of a common rice " Jiahua-1 " are constructed binary
Expression vector turns Agrobacterium, and the cDNA sequence is transferred to containing a seedling stage Huang by the transgenic technology of mediated by agriculture bacillus
Phyllopodium because plant in, offspring's Plant Leaf color turns green, and illustrating that the equal sites of the gene in common rice are destroyed can lead to
Early stage rice Development of Chloroplasts is obstructed, to make rice seedling yellow leaf.The rice seedling yellow leaf obtained through the invention
Gene can be used for studying the photosynthesis of higher plant, the biosynthesis of chlorophyll, the genetic variation and genetic differentiation of chloroplaset and development etc., and
And can be used as mark property in breeding of hybrid rice, in the purity of sterile line of hybridized rice itself and its preparing hybrid rice
With application.
The invention has the benefit that obtaining a rice seedling yellow leaf gene and quickly detecting whether to contain the base
The detection method of cause, and using this method can fast slowdown monitoring contain the rice plant of the gene, detection accuracy is high, behaviour
Make simply, it is low in cost.So that if the allele is destroyed or can lead to by the yellow leaf gene substitution in common rice
The discovery in advance of rice seedling yellow leaf character such as, prevents in advance.The rice seedling yellow leaf base obtained additionally by the present invention
Because can be used for Chlorophyll synthesis, Development of Chloroplasts and photosynthetic mechanism of Study On Rice etc., and it can also be used as mark property
It is applied in the purity of breeding of hybrid rice, sterile line of hybridized rice itself and its preparing hybrid rice.
Detailed description of the invention
Fig. 1 is common rice and the Plant Leaf colour contrast figure containing seedling stage yellow leaf trans-genetic hybrid rice；
Fig. 2 is that specific dCAPS PCR primer of the invention carries out PCR amplification result to the gene.T1 represents light water
Rice pcr amplification product, T2, which is represented, contains yellow leaf trans-genetic hybrid rice pcr amplification product；
Fig. 3 is that the present invention carries out pcr amplification productEam1105I digestion testing result.T3 represents T1 processEamSegment after 1105I digestion；T4 represents T2 processEamSegment after 1105I digestion；
Fig. 4 is pTCK303 carrier structure schematic diagram of the invention；
Fig. 5 is the electrophoretogram of pTCK303 binary vector of the invention；
Fig. 6 is the electrophoretogram that pTCK303 binary vector of the invention carries out digestion verification.
Specific embodiment
1 paddy DNA of embodiment extracts
1, the fresh Rice Seedlings blade of 1.0g is weighed, blade is cut into the segment of 0.5cm long, is put into the mortar of pre-cooling
In, liquid nitrogen grinding is added to powdered, 0.5ml is taken to be transferred in 2.0ml centrifuge tube.
2,2 × CTAB of 65 DEG C of 1ml preheatings is added into 2.0ml centrifuge tube, acutely rocks mixing, rear 65 DEG C of water-bath 40-
During which 60min takes out concussion for several times (at least 3 times).
3, centrifuge tube after the water bath is over, is taken out, is cooled to room temperature, isometric chloroform-isoamyl alcohol (24:1) mixed liquor is added,
It mixes well, is centrifuged 10min under conditions of 9000 revs/min.
4, Aspirate supernatant is added the isopropanol of its 2/3 volume, mixes well, nucleic acid is precipitated, is placed at room temperature for 10min,
It is centrifuged 10min under conditions of 12000 revs/min, precipitates nucleic acid.
5, abandon supernatant, with 70% ethanol washing precipitate 2 times (every time plus 70% ethyl alcohol 500 μ l, under conditions of 12000 revs/min from
Heart 3min), it dries later.
6, it is dissolved in after drying at room temperature in the sterile water or TE buffer of 100-200 μ l.
7, take 2 μ l for subsequent pcr amplification reaction.
2 PCR amplification of embodiment and genetic test
(1) common rice gene and a rice seedling yellow leaf gene are obtained respectively
1,50 μ L PCR reaction systems are as follows: 25 μ l 2 × PCR buffer for KOD FX；10μl 2Mm dNTPs；3
10 μM of primers of μ l；3μl Template DNA；8μl ddH2O；1 μ l KOD FX(1.0 U/ μ l).
Primer sequence are as follows:
F:5'-CCCCCGGGIt is restriction enzyme at ACATTCTCCTTCGTCTACCTCCGTC-3'(scribing lineSmaI alkali
Base identifies sequence)
R:5'-GCGTCGACIt is restriction enzyme at ATAGCCCACTGCTCTTTTCTTTCAT-3'(scribing lineSalI alkali
Base identifies sequence)
2, pcr amplification reaction carries out in PCR amplification instrument.
Temperature and reaction condition are as follows: 94 DEG C of initial denaturation 2min, 98 DEG C of denaturation 10sec, 60 DEG C of annealing 30sec, 68 DEG C extend
6min, 35 circulations；Last 68 DEG C of extensions 6min.
Pcr amplification product length is 6.0kbp, the common rice gene sequencing result such as SEQ ID No.7, seedling stage yellow leaf
The rice gene sequencing result such as SEQ ID No.1.
Referring to the gene ORF sequence B ioxm software translation obtained on the website KOME at amino acid sequence common rice such as
Shown in SEQ ID No.8, seedling stage yellow leaf rice is as shown in SEQ ID No.2.
(2) a rice seedling yellow leaf gene is detected
1,20 μ L PCR reaction systems are as follows: 2 μ l template DNAs；1 10 μM of μ l primer；2.5μl buffer forTaq
DNAPol；0.5μl dNTP；1μl DMSO；12μl ddH2O；1μlTaqEnzyme (5U/ μ l).
The nucleotide sequence of primer is as shown in SEQ ID No.3 and SEQ ID No.4:
Upstream: 5 '-GTGTGTACTCTCAGATGAAGTACTTGGACTCTCAGT-3 '
Downstream: 5 '-CACATTGTATTTGCAGAACA-3 '.
2, pcr amplification reaction carries out in PCR amplification instrument.
Temperature and reaction condition are as follows: 94 DEG C of initial denaturation 4min, 94 DEG C of denaturation 30sec, 60 DEG C of annealing 30sec, 72 DEG C are prolonged
Stretch 90sec, 35 circulations；Last 72 DEG C of extensions 6min.
Obtained amplified production is splined on 4.0% Ago-Gel simultaneously electrophoresis, in UVP gel after ethidium bromide staining
It is imaged on imager, as a result as shown in (Fig. 2), 115bp band (Fig. 2) occurs in common rice, is one in SEQ ID No.7
Section；There is 113bp band (Fig. 2) containing a rice seedling yellow leaf gene, is one section in SEQ ID No.1.
The verifying of 3 gene function of embodiment
Using60Co gamma-ray irradiation or other physical method mutagenesis destroy the gene order of SEQ ID No.7 in rice
Obtain the gene order of SEQ ID No.1.Gene of the gene order of SEQ ID No.1 relative to SEQ ID No.7 is sequenced
There are two the missing of nucleotide (TG), rna levels to reduce for sequence, then leads to the rice seedling plant yellow leaf.The function of the gene
RNA interference experiment is the result shows that the Progeny plants leaf color containing the normal gene is in green.
1, the positioning that rice seedling yellow leaf gene is carried out using SSR and InDel molecular labeling passes through sequencing and sxemiquantitative
RT-PCR technology shows that the gene order changes, the expression quantity decline of rna level.
2, in rice genome library website (http://rice.plantbiology.msu.edu/cgi-bin/
Gbrowse/rice/ DNA sequence dna, cDNA sequence and the amino acid sequence that the target gene is found on), utilize website NCBI
(http://blast.ncbi.nlm.nih.gov/Blast.cgi?PROGRAM=blastp&PAGE_TYPE=
BlastSearch&LINK_LOC=blasthome) and pfam (http://pfam.xfam.org/) target gene encoded
Protein structure domain carries out forecast analysis, based on the analysis results, then by design of primers in the gene location where structural domain, then sharp
With 5.0 design primer of primer-design software Primer, design of primers fragment length is about 300-400bp, and annealing temperature exists
Between 55-58 DEG C.And restriction enzyme site verifying need to be carried out to the segment of amplification, determine it is errorless after, primer upstream and downstream be added enzyme
Enzyme site.
Primer sequence:
F1:5'-CGAGCTCIt is restriction enzyme at TAGAAATGTCGTCTGAGCCTT-3'(scribing lineSacI base is known
Other sequence)
R1:5'-GGACTAGTIt is restriction enzyme at CGCTGATACTCCACAGAACGC-3'(scribing lineSpeI base is known
Other sequence)
F2:5'-CGGGATCCIt is restriction enzyme at TAGAAATGTCGTCTGAGCCTT-3'(scribing lineBamHI base
Identify sequence)
R2:5'-GGGGTACCIt is restriction enzyme at CGCTGATACTCCACAGAACGC-3'(scribing lineKpnI base is known
Other sequence)
3, pTCK303 binary vector is constructed
(1) using common rice such as " Jiahua-1 " cDNA as template, target fragment is expanded and is recycled, pMD18T is then connected
The plasmid that sequencing is correctly cloned is extracted in carrier, the sequencing of picking colony PCR positive colony sample presentation.
(2) T-1 plasmid (connect T for F1/R1 amplified production and carry positive colony) and pTCK303(Fig. 4) is carried out respectivelySacI andSpeI double digestion connects conversion after recycling target fragment, selects positive colony, extracts plasmid (Fig. 5).
(3) by T-2 plasmid (by F2/R2 amplified production connect T carry positive colony) and step 2 upgrading grain recycle, distinguish
It carries outBamHI andKpnI double digestion.Conversion is connected after recycling target fragment, the positive colony selected is exactly final interference table
Up to carrier.
(4) digestion verification (Fig. 6) is carried out to gained pTCK303 binary vector, digestion post-fragment position and purpose base
Cause it is in the same size, illustrate in step 3 that there is no problem for gained positive colony.
(5) transgenic technology for utilizing mediated by agriculture bacillus, is transferred to Agrobacterium competence for pTCK303 binary vector, converts
The callus of the rice of seedling stage yellow leaf gene, the positive first generation transgenic plant of acquisition, leaf color is in green.
4, the seed of first generation transgenic plant is sowed, leaf color separates, and in green and yellow leaf phenotype, and meets
Mendelian inheritance segregation ratio, that is, 3:1.
5, the gene normal expression is rice plant leaf color in green, and the mutant plant seedling leaf color that its expression is lowered is in
Yellow leaf phenotype, thus can be identified by this character mutation a plant whether the gene containing a rice seedling yellow.
Sequence table
SEQUENCE LISTING
<110>Shanghai Normal University
The rapid detection method of<120>rice seedling yellow leaf genes
<130> none
<160> 5
<170> PatentIn version 3.3
<210> 1
<211> 5941
<212> DNA
<213>rice
<400> 1
atgctcctcc tcttcctccc ccacccttcc ccacctctcc tcccccccgc cgcctcgaga 60
acccgacccc cgccgcgcct cctcctccct cccatccacg cctccccctc gccggagctc 120
ctcgccaagt cggctctccg ccgcatctcc gacaagctcc gcagcctcgg ctacctcgag 180
gccgaccacc cggaggcggc ccccggtccg gcggcgccgg aggcgggcgc cggcgcgtcc 240
cccggcgaga tattcgttcc cacgccggcc cagctcccgc gccaccgcgt cggctccacc 300
ctagacccga gctgggccac gggcgatggg gagggggccg ccgcgtctcg ccgcaggagg 360
cgcggcggga gggatagtag cgccgcggcg tcggcgccgc cgtccgccgc ggagctggcg 420
ctgccgaggg acgagctgcg gcggcttcag ggggccggga ttcggctgcg gaacaggctc 480
aaggtcggga aggccggggt gacggagggg atcgtgaacg ggatacacga gcggtggcgc 540
aacgcggagc tcgtgaagat caggtgcgac gacgtctccg ccatgaacat gaagcgcacg 600
cacgagattc tcgaggtgag cgctgccccg cgcatatgat aatcgagtcc agcgtaggtt 660
tgtagctgtg cgcctgtgcc tcgcctagta agtaaaacgt tcagaaaatt tggatgccgg 720
aaggcatcgg agtgctacta tatgttgaaa ttgcacagag tattagttgt atgttaattc 780
tgaatttcat cgcacaatgt gacagggtaa gatgctgagt atagtaatat agtgctagca 840
tatgattgtg cccacttcgg aagtccctgt tagcatggaa atggcagcat gtttattcaa 900
aagggataca actttgaagt gatcaaggat tcaagatagg gatattttat ttttaattgt 960
ttgaaatgaa tccatttagc agtgtactgg catgcacata atcattcctt cttcgaagca 1020
ttttattata ccaaattcca aggcatttct tggattttct cttaaagtaa tctatctcaa 1080
tttttgtttt agggcatttc tatctcaacg cttttcagtt tgcaattgac atgctcaatg 1140
tgttttttta tagagaaaaa ctggaggctt agtcatttgg agatctggaa gcaccatcat 1200
attatataga ggaactgact ataagtatcc ttacttccat gatagagaaa tgaagaatga 1260
catggatgag tcatcagagc atacaagttc tgatgacgag gacgcggatc ttgctataat 1320
agcttccgaa caaagtggca gtgaagagga ttctgataat cctgcagaac atggtagcaa 1380
tcatactgag gagggcgatg atcttactag acggtttggt gttgatgcgc ttgagggaaa 1440
tttagacatt ggatctgcag agcaaagcat taacagtgct acaaaagatc agcaagcaat 1500
attacataca agcactaatg tgtctcgtcc tagcgagatc agtggtcgag ctcggtcaac 1560
cttagttgct ggtgttggtt ctccaaataa gtttcggtta caattacctg gggaggtcaa 1620
actcgcagag gaagctgaca aattattgga tgggttgggt ccacggtttt ctgattggtg 1680
ggggtatgac cctcttccag tggatgctga tttgttgcca gcaattgttc ctggataccg 1740
taggcctttt cgtctgcttc cttcaggagt accaccgagg ttaactgata gagaaatgac 1800
aatccttagg agactagctc gtcccctacc ctaccattac gcacttggta atttctagtt 1860
atttgtcttg tataatatct ttcatggctt tgactttgat aaggaccact gttatttact 1920
ttcgtggtgt catttatcta ggacgaagta gtaaccttca agggttggct gcatctatga 1980
tcaaactatg ggaaagatgt gaggtagcaa aggttgcaat taagagaggt gcagagaaca 2040
ttgatagtga tctcatttct gagaaactga aggtaggttg gtaaagaacc cagctatgag 2100
gaattaattg tagatatgta attattcata gcatttgaca tttctcgagc attcttgcga 2160
taaaacattg atgacattac agatttatct ttcacgtgtc atcgaatttt gaaatgtaaa 2220
tcatttgcct gtattcctaa tttcctatca taattgaagt tgaaccatag cagcccagaa 2280
tttttacatg ttgaatcaaa atcacttctt tttatgatat atgttccctc atactcatgc 2340
atattttaca tgcagtatga aggcacttaa caatagattc atatttttta tgtgtcgcct 2400
tgatttatgt ggcaccaaat attaggaaaa tacactttag tatctctttc tgatgtgaac 2460
catttcagca caggctccag gccttcaacc tattatatcc taaactttcc attatgagaa 2520
attatcatta caatcatcag tttcaaaaga aaatcagaga ggaggggggg ggggggttat 2580
gttccaccat gacattctta atcttgcagg gcttgacagg tgggaccctt ttatcaaggg 2640
ataatgagtc aattgtattc tacagaggga aagatttcct gcccacagct gtctctcttg 2700
caatagaaaa gcgtagaaag tatggcaatt ctacaatcag taaccctaag cttaattttg 2760
acaaaagcac accgcaaaat tcttctaaac tgaagatggc tactgatgtc tcactagatg 2820
ggcatgaatg ctatgaaaag aagcataaag atgaaactgc agtttcagat aatagagcag 2880
aatctcttaa tgtgtttgcc cagaatgttg aggccagatt atctcaggtt tgcttttctt 2940
ttctagtagt gccatctctt gataattttg atttggtagt ttgaaaagaa aaacacttga 3000
aatagttaaa ttcatataga aatattgttc aggcaattgc agagaaagag aagactgaaa 3060
agcttattga agagctagaa atgtcgtctg agccttctag agctgaaact agagaggtta 3120
tctcagaaga ggagaggtac atgttgagga aagttggttt aaagatgaaa tcattcttat 3180
tgctaggtta gtattctcct caaccttgta ttgtgcagca tgcatctgtt cagaagtcaa 3240
aatgctacct aatttgacta ccaaattagt tataggaaag tagattacca gacagtgttc 3300
tccttggtga cccctaggca ttcggtgtcc gctccgccta ggtcagcaag ctgatttttc 3360
ccgcctagga agcagttagg cactaggtcc cacctatggc cgtgccatgt gcctactgcc 3420
tagcaccaag ggaaacactt ctatcagact gtaatattac cacttctaga gttttcatct 3480
gtttagctca taccaagtgg cattttaatc cttgtaaatc ctttttgtgg agacagtgtg 3540
agtattatgt ggttttttgt tactttggga gtgcacatat ttgaattagt tcggtcatct 3600
gttgtgattc atcgcataac ctccttttgt accattctat acctgtacat gatttgacta 3660
ttattgcctt tccaagcagt agcatccgca gccctaaggt caataaacaa aaagagagcg 3720
cgacataccc atggactcta tacaaactgt tctttctgtg atttctgcac gatattgata 3780
cctcattcgc ttgtatctta ctgtttttta ttatttcttt aattcaacat cctttttttt 3840
ttctattgat tgcaggaagg agaggagttt ttgatggaac tgttgaaaat atgcaccttc 3900
actggaagta ccgagaactt gtcaaaatta tatgtaaaga acacaacatt aaagatgttg 3960
aatatgctgc tcgaactcta gaggctgaga gtggtggtat attggtggca gtggaaagag 4020
tgagcaaagc acatgcaatt ataatatatc ggggaaagaa ctatcagagg ccatcaactc 4080
tcaggccaaa atctttgtta aataaaaagg atgctttgaa gcgttctgtg gagtatcagc 4140
gttataaggt gaaactgaaa cctattagct tgctgtggtt cacttgtgga gttatttttt 4200
agatctcaaa ttaatattcc ttgtttattt tagtccctga agcttcatgt gcttaatctt 4260
tcaaagaata tagactactt gaaggatcag atggtgagtt ttttgttcaa tgctgagcct 4320
tttccattcg ttttgttgta aatgctatta ttaatttact gatgatgctt cagtttttta 4380
aacaaatgga ggttcaacct gttactccaa ctaatggaat gaactcaggc caccacaacc 4440
aaggcattct tgatctcaat gttaattctg gaacattagt ggacaaaaaa gaagtaaatt 4500
ttattatctt aactgaagtt ccattatggc tcaatatatt caatcatgtc tgattatctg 4560
tttattaaat atatatggtc aaggaagtgt ctgaagttct tcctgagtgt gccaagtcag 4620
tggttgtaga gtgttcttca ggtgaaagtg aaacagaggg aacctcagtt ttaaccaaat 4680
ctggagttcc cttggatgtt atgcaggttg ctggtcctcc cattccttta cgaaaattct 4740
tgtgtgtttt gattcttcgc aacaattttg tggtggacta atggttcatt gcacattgta 4800
tttgcagaac aagttgttat gtttcagcaa gcatacagat gatctgtctg aaactacaag 4860
ttcctgtcac tgagagtaca agtacttcat ctgagagtac acacgtgagt ttattgttga 4920
tttttcccag tgcatggcaa ttattttctg ctgatgctga tgatttgact gcagcaaagt 4980
ccacttagtt cttctgtgat gcacaattca gatagtcaca gagtaagtgg ctcaaaattt 5040
gtaggaaccc ttacacctgt acacgagctg aaattggatg agaaatcttc tcagttgcct 5100
tccgcagctg ctccactttc aaaccgagaa aggctcatgc taaggaagca agctctgaaa 5160
atgaagaaac ggccagtact cgctgttggt aagtctttgt ccccaataca gtttacacca 5220
gatttcaatt aatatgctga gcatccgatt cctttcatag ggaggaacaa tgtcatcaca 5280
ggagtcgcaa aagcaataaa aacacacttc aagaagcatc ctcttgctat tgtgaatatc 5340
aagaaccgag cagacggaac tccaattcag cagttgattt cagaactaga ggtttgtata 5400
gtctataaca gggagtagtt caccccaaaa atactgaatc aatctgaaat tctatattgt 5460
tcaatggaat caggaagcaa ctggatcggt tctagtgtcg cgggagccca acaaagtgat 5520
tttgtacagg ggttggggag cagatgtggc ccagaatagc ttaagtggaa ataatagtac 5580
agaacaagtt gaaaaggagg tgatatcacc tcagctcctg gaagccgtca gattagaatg 5640
tggattacac cctggtgaat cagagtagat ggatctctag atcctgtaga tggattatct 5700
tcatcgaatt gcagttggtg aagttgtgct cgcgccttca atgtccagat tgcgttttcc 5760
ctcgtcgttt ctgaccacag ttttctggtt ggcctcctct tgacacagcc cgttgacgcg 5820
ctgttcaaat tggaatgtag acacgaacat tgttttatct gtagtgtctc cttgtcacct 5880
tcctttgtaa atctaaaaca tctgaaaggc ctaactggat aaatattcct gcaacctaaa 5940
a5941
<210> 2
<211> 821
<212> PRT
<213>rice
<400> 2
Met Leu Leu Leu Phe Leu Pro His Pro Ser Pro Pro Leu Leu Pro Pro
1 5 10 15
Ala Ala Ser Arg Thr Arg Pro Pro Pro Arg Leu Leu Leu Pro Pro Ile
20 25 30
His Ala Ser Pro Ser Pro Glu Leu Leu Ala Lys Ser Ala Leu Arg Arg
35 40 45
Ile Ser Asp Lys Leu Arg Ser Leu Gly Tyr Leu Glu Ala Asp His Pro
50 55 60
Glu Ala Ala Pro Gly Pro Ala Ala Pro Glu Ala Gly Ala Gly Ala Ser
65 70 75 80
Pro Gly Glu Ile Phe Val Pro Thr Pro Ala Gln Leu Pro Arg His Arg
85 90 95
Val Gly Ser Thr Leu Asp Pro Ser Trp Ala Thr Gly Asp Gly Glu Gly
100 105 110
Ala Ala Ala Ser Arg Arg Arg Arg Arg Gly Gly Arg Asp Ser Ser Ala
115 120 125
Ala Ala Ser Ala Pro Pro Ser Ala Ala Glu Leu Ala Leu Pro Arg Asp
130 135 140
Glu Leu Arg Arg Leu Gln Gly Ala Gly Ile Arg Leu Arg Asn Arg Leu
145 150 155 160
Lys Val Gly Lys Ala Gly Val Thr Glu Gly Ile Val Asn Gly Ile His
165 170 175
Glu Arg Trp Arg Asn Ala Glu Leu Val Lys Ile Arg Cys Asp Asp Val
180 185 190
Ser Ala Met Asn Met Lys Arg Thr His Glu Ile Leu Glu Arg Lys Thr
195 200 205
Gly Gly Leu Val Ile Trp Arg Ser Gly Ser Thr Ile Ile Leu Tyr Arg
210 215 220
Gly Thr Asp Tyr Lys Tyr Pro Tyr Phe His Asp Arg Glu Met Lys Asn
225 230 235 240
Asp Met Asp Glu Ser Ser Glu His Thr Ser Ser Asp Asp Glu Asp Ala
245 250 255
Asp Leu Ala Ile Ile Ala Ser Glu Gln Ser Gly Ser Glu Glu Asp Ser
260 265 270
Asp Asn Pro Ala Glu His Gly Ser Asn His Thr Glu Glu Gly Asp Asp
275 280 285
Leu Thr Arg Arg Phe Gly Val Asp Ala Leu Glu Gly Asn Leu Asp Ile
290 295 300
Gly Ser Ala Glu Gln Ser Ile Asn Ser Ala Thr Lys Asp Gln Gln Ala
305 310 315 320
Ile Leu His Thr Ser Thr Asn Val Ser Arg Pro Ser Glu Ile Ser Gly
325 330 335
Arg Ala Arg Ser Thr Leu Val Ala Gly Val Gly Ser Pro Asn Lys Phe
340 345 350
Arg Leu Gln Leu Pro Gly Glu Val Lys Leu Ala Glu Glu Ala Asp Lys
355 360 365
Leu Leu Asp Gly Leu Gly Pro Arg Phe Ser Asp Trp Trp Gly Tyr Asp
370 375 380
Pro Leu Pro Val Asp Ala Asp Leu Leu Pro Ala Ile Val Pro Gly Tyr
385 390 395 400
Arg Arg Ser Ser Asn Leu Gln Gly Leu Ala Ala Ser Met Ile Lys Leu
405 410 415
Trp Glu Arg Cys Glu Val Ala Lys Val Ala Ile Lys Arg Gly Ala Glu
420 425 430
Asn Ile Asp Ser Asp Leu Ile Ser Glu Lys Leu Lys Gly Leu Thr Gly
435 440 445
Gly Thr Leu Leu Ser Arg Asp Asn Glu Ser Ile Val Phe Tyr Arg Gly
450 455 460
Lys Asp Phe Leu Pro Thr Ala Val Ser Leu Ala Ile Glu Lys Arg Arg
465 470 475 480
Lys Tyr Gly Asn Ser Thr Ile Ser Asn Pro Lys Leu Asn Phe Asp Lys
485 490 495
Ser Thr Pro Gln Asn Ser Ser Lys Leu Lys Met Ala Thr Asp Val Ser
500 505 510
Leu Asp Gly His Glu Cys Tyr Glu Lys Lys His Lys Asp Glu Thr Ala
515 520 525
Val Ser Asp Asn Arg Ala Glu Ser Leu Asn Val Phe Ala Gln Asn Val
530 535 540
Glu Ala Arg Leu Ser Gln Ala Ile Ala Glu Lys Glu Lys Thr Glu Lys
545 550 555 560
Leu Ile Glu Glu Leu Glu Met Ser Ser Glu Pro Ser Arg Ala Glu Thr
565 570 575
Arg Glu Val Ile Ser Glu Glu Glu Arg Tyr Met Leu Arg Lys Val Gly
580 585 590
Leu Lys Met Lys Ser Phe Leu Leu Leu Gly Arg Arg Gly Val Phe Asp
595 600 605
Gly Thr Val Glu Asn Met His Leu His Trp Lys Tyr Arg Glu Leu Val
610 615 620
Lys Ile Ile Cys Lys Glu His Asn Ile Lys Asp Val Glu Tyr Ala Ala
625 630 635 640
Arg Thr Leu Glu Ala Glu Ser Gly Gly Ile Leu Val Ala Val Glu Arg
645 650 655
Val Ser Lys Ala His Ala Ile Ile Ile Tyr Arg Gly Lys Asn Tyr Gln
660 665 670
Arg Pro Ser Thr Leu Arg Pro Lys Ser Leu Leu Asn Lys Lys Asp Ala
675 680 685
Leu Lys Arg Ser Val Glu Tyr Gln Arg Tyr Lys Ser Leu Lys Leu His
690 695 700
Val Leu Asn Leu Ser Lys Asn Ile Asp Tyr Leu Lys Asp Gln Met Phe
705 710 715 720
Phe Lys Gln Met Glu Val Gln Pro Val Thr Pro Thr Asn Gly Met Asn
725 730 735
Ser Gly His His Asn Gln Gly Ile Leu Asp Leu Asn Val Asn Ser Gly
740 745 750
Thr Leu Val Asp Lys Lys Glu Glu Val Ser Glu Val Leu Pro Glu Cys
755 760 765
Ala Lys Ser Val Val Val Glu Cys Ser Ser Gly Glu Ser Glu Thr Glu
770 775 780
Gly Thr Ser Val Leu Thr Lys Ser Gly Val Pro Leu Asp Val Met Gln
785 790 795 800
Asn Lys Leu Leu Cys Phe Ser Lys His Thr Asp Asp Leu Ser Glu Thr
805 810 815
Thr Ser Ser Cys His
820
<210> 3
<211> 36
<212> DNA
<213>artificial sequence
<400> 3
gtgtgtactc tcagatgaag tacttggact ctcagt
36
<210> 4
<211> 20
<212> DNA
<213>artificial sequence
<400> 4
cacattgtat ttgcagaaca
20
<210> 5
<211> 115
<212> DNA
<213>rice
<400> 5
gtgtgtactc tcagatgaag tacttgtact ctcagtcaga caggaacttg tagtttcaga 60
cagatcatct gtatgcttgc tgaaacataa caacttgttc tgcaaataca atgtg 115
<210> 6
<211> 113
<212> DNA
<213>rice
<400> 6
gtgtgtactc tcagatgaag tacttgtact ctcagtgaca ggaacttgta gtttcagaca60
gatcatctgt atgcttgctg aaacataaca acttgttctg caaatacaat gtg 113
<210> 7
<211> 5943
<212> DNA
<213>rice
<400> 7
atgctcctcc tcttcctccc ccacccttcc ccacctctcc tcccccccgc cgcctcgaga 60
acccgacccc cgccgcgcct cctcctccct cccatccacg cctccccctc gccggagctc 120
ctcgccaagt cggctctccg ccgcatctcc gacaagctcc gcagcctcgg ctacctcgag 180
gccgaccacc cggaggcggc ccccggtccg gcggcgccgg aggcgggcgc cggcgcgtcc 240
cccggcgaga tattcgttcc cacgccggcc cagctcccgc gccaccgcgt cggctccacc 300
ctagacccga gctgggccac gggcgatggg gagggggccg ccgcgtctcg ccgcaggagg 360
cgcggcggga gggatagtag cgccgcggcg tcggcgccgc cgtccgccgc ggagctggcg 420
ctgccgaggg acgagctgcg gcggcttcag ggggccggga ttcggctgcg gaacaggctc 480
aaggtcggga aggccggggt gacggagggg atcgtgaacg ggatacacga gcggtggcgc 540
aacgcggagc tcgtgaagat caggtgcgac gacgtctccg ccatgaacat gaagcgcacg 600
cacgagattc tcgaggtgag cgctgccccg cgcatatgat aatcgagtcc agcgtaggtt 660
tgtagctgtg cgcctgtgcc tcgcctagta agtaaaacgt tcagaaaatt tggatgccgg 720
aaggcatcgg agtgctacta tatgttgaaa ttgcacagag tattagttgt atgttaattc 780
tgaatttcat cgcacaatgt gacagggtaa gatgctgagt atagtaatat agtgctagca 840
tatgattgtg cccacttcgg aagtccctgt tagcatggaa atggcagcat gtttattcaa 900
aagggataca actttgaagt gatcaaggat tcaagatagg gatattttat ttttaattgt 960
ttgaaatgaa tccatttagc agtgtactgg catgcacata atcattcctt cttcgaagca 1020
ttttattata ccaaattcca aggcatttct tggattttct cttaaagtaa tctatctcaa 1080
tttttgtttt agggcatttc tatctcaacg cttttcagtt tgcaattgac atgctcaatg 1140
tgttttttta tagagaaaaa ctggaggctt agtcatttgg agatctggaa gcaccatcat 1200
attatataga ggaactgact ataagtatcc ttacttccat gatagagaaa tgaagaatga 1260
catggatgag tcatcagagc atacaagttc tgatgacgag gacgcggatc ttgctataat 1320
agcttccgaa caaagtggca gtgaagagga ttctgataat cctgcagaac atggtagcaa 1380
tcatactgag gagggcgatg atcttactag acggtttggt gttgatgcgc ttgagggaaa 1440
tttagacatt ggatctgcag agcaaagcat taacagtgct acaaaagatc agcaagcaat 1500
attacataca agcactaatg tgtctcgtcc tagcgagatc agtggtcgag ctcggtcaac 1560
cttagttgct ggtgttggtt ctccaaataa gtttcggtta caattacctg gggaggtcaa 1620
actcgcagag gaagctgaca aattattgga tgggttgggt ccacggtttt ctgattggtg 1680
ggggtatgac cctcttccag tggatgctga tttgttgcca gcaattgttc ctggataccg 1740
taggcctttt cgtctgcttc cttcaggagt accaccgagg ttaactgata gagaaatgac 1800
aatccttagg agactagctc gtcccctacc ctaccattac gcacttggta atttctagtt 1860
atttgtcttg tataatatct ttcatggctt tgactttgat aaggaccact gttatttact 1920
ttcgtggtgt catttatcta ggacgaagta gtaaccttca agggttggct gcatctatga 1980
tcaaactatg ggaaagatgt gaggtagcaa aggttgcaat taagagaggt gcagagaaca 2040
ttgatagtga tctcatttct gagaaactga aggtaggttg gtaaagaacc cagctatgag 2100
gaattaattg tagatatgta attattcata gcatttgaca tttctcgagc attcttgcga 2160
taaaacattg atgacattac agatttatct ttcacgtgtc atcgaatttt gaaatgtaaa 2220
tcatttgcct gtattcctaa tttcctatca taattgaagt tgaaccatag cagcccagaa 2280
tttttacatg ttgaatcaaa atcacttctt tttatgatat atgttccctc atactcatgc 2340
atattttaca tgcagtatga aggcacttaa caatagattc atatttttta tgtgtcgcct 2400
tgatttatgt ggcaccaaat attaggaaaa tacactttag tatctctttc tgatgtgaac 2460
catttcagca caggctccag gccttcaacc tattatatcc taaactttcc attatgagaa 2520
attatcatta caatcatcag tttcaaaaga aaatcagaga ggaggggggg ggggggttat 2580
gttccaccat gacattctta atcttgcagg gcttgacagg tgggaccctt ttatcaaggg 2640
ataatgagtc aattgtattc tacagaggga aagatttcct gcccacagct gtctctcttg 2700
caatagaaaa gcgtagaaag tatggcaatt ctacaatcag taaccctaag cttaattttg 2760
acaaaagcac accgcaaaat tcttctaaac tgaagatggc tactgatgtc tcactagatg 2820
ggcatgaatg ctatgaaaag aagcataaag atgaaactgc agtttcagat aatagagcag 2880
aatctcttaa tgtgtttgcc cagaatgttg aggccagatt atctcaggtt tgcttttctt 2940
ttctagtagt gccatctctt gataattttg atttggtagt ttgaaaagaa aaacacttga 3000
aatagttaaa ttcatataga aatattgttc aggcaattgc agagaaagag aagactgaaa 3060
agcttattga agagctagaa atgtcgtctg agccttctag agctgaaact agagaggtta 3120
tctcagaaga ggagaggtac atgttgagga aagttggttt aaagatgaaa tcattcttat 3180
tgctaggtta gtattctcct caaccttgta ttgtgcagca tgcatctgtt cagaagtcaa 3240
aatgctacct aatttgacta ccaaattagt tataggaaag tagattacca gacagtgttc 3300
tccttggtga cccctaggca ttcggtgtcc gctccgccta ggtcagcaag ctgatttttc 3360
ccgcctagga agcagttagg cactaggtcc cacctatggc cgtgccatgt gcctactgcc 3420
tagcaccaag ggaaacactt ctatcagact gtaatattac cacttctaga gttttcatct 3480
gtttagctca taccaagtgg cattttaatc cttgtaaatc ctttttgtgg agacagtgtg 3540
agtattatgt ggttttttgt tactttggga gtgcacatat ttgaattagt tcggtcatct 3600
gttgtgattc atcgcataac ctccttttgt accattctat acctgtacat gatttgacta 3660
ttattgcctt tccaagcagt agcatccgca gccctaaggt caataaacaa aaagagagcg 3720
cgacataccc atggactcta tacaaactgt tctttctgtg atttctgcac gatattgata 3780
cctcattcgc ttgtatctta ctgtttttta ttatttcttt aattcaacat cctttttttt 3840
ttctattgat tgcaggaagg agaggagttt ttgatggaac tgttgaaaat atgcaccttc 3900
actggaagta ccgagaactt gtcaaaatta tatgtaaaga acacaacatt aaagatgttg 3960
aatatgctgc tcgaactcta gaggctgaga gtggtggtat attggtggca gtggaaagag 4020
tgagcaaagc acatgcaatt ataatatatc ggggaaagaa ctatcagagg ccatcaactc 4080
tcaggccaaa atctttgtta aataaaaagg atgctttgaa gcgttctgtg gagtatcagc 4140
gttataaggt gaaactgaaa cctattagct tgctgtggtt cacttgtgga gttatttttt 4200
agatctcaaa ttaatattcc ttgtttattt tagtccctga agcttcatgt gcttaatctt 4260
tcaaagaata tagactactt gaaggatcag atggtgagtt ttttgttcaa tgctgagcct 4320
tttccattcg ttttgttgta aatgctatta ttaatttact gatgatgctt cagtttttta 4380
aacaaatgga ggttcaacct gttactccaa ctaatggaat gaactcaggc caccacaacc 4440
aaggcattct tgatctcaat gttaattctg gaacattagt ggacaaaaaa gaagtaaatt 4500
ttattatctt aactgaagtt ccattatggc tcaatatatt caatcatgtc tgattatctg 4560
tttattaaat atatatggtc aaggaagtgt ctgaagttct tcctgagtgt gccaagtcag 4620
tggttgtaga gtgttcttca ggtgaaagtg aaacagaggg aacctcagtt ttaaccaaat 4680
ctggagttcc cttggatgtt atgcaggttg ctggtcctcc cattccttta cgaaaattct 4740
tgtgtgtttt gattcttcgc aacaattttg tggtggacta atggttcatt gcacattgta 4800
tttgcagaac aagttgttat gtttcagcaa gcatacagat gatctgtctg aaactacaag 4860
ttcctgtctg actgagagta caagtacttc atctgagagt acacacgtga gtttattgtt 4920
gatttttccc agtgcatggc aattattttc tgctgatgct gatgatttga ctgcagcaaa 4980
gtccacttag ttcttctgtg atgcacaatt cagatagtca cagagtaagt ggctcaaaat 5040
ttgtaggaac ccttacacct gtacacgagc tgaaattgga tgagaaatct tctcagttgc 5100
cttccgcagc tgctccactt tcaaaccgag aaaggctcat gctaaggaag caagctctga 5160
aaatgaagaa acggccagta ctcgctgttg gtaagtcttt gtccccaata cagtttacac 5220
cagatttcaa ttaatatgct gagcatccga ttcctttcat agggaggaac aatgtcatca 5280
caggagtcgc aaaagcaata aaaacacact tcaagaagca tcctcttgct attgtgaata 5340
tcaagaaccg agcagacgga actccaattc agcagttgat ttcagaacta gaggtttgta 5400
tagtctataa cagggagtag ttcaccccaa aaatactgaa tcaatctgaa attctatatt 5460
gttcaatgga atcaggaagc aactggatcg gttctagtgt cgcgggagcc caacaaagtg 5520
attttgtaca ggggttgggg agcagatgtg gcccagaata gcttaagtgg aaataatagt 5580
acagaacaag ttgaaaagga ggtgatatca cctcagctcc tggaagccgt cagattagaa 5640
tgtggattac accctggtga atcagagtag atggatctct agatcctgta gatggattat 5700
cttcatcgaa ttgcagttgg tgaagttgtg ctcgcgcctt caatgtccag attgcgtttt 5760
ccctcgtcgt ttctgaccac agttttctgg ttggcctcct cttgacacag cccgttgacg 5820
cgctgttcaa attggaatgt agacacgaac attgttttat ctgtagtgtc tccttgtcac 5880
cttcctttgt aaatctaaaa catctgaaag gcctaactgg ataaatattc ctgcaaccta 5940
aaa 5943
<210> 8
<211> 1012
<212> PRT
<213>rice
<400> 8
Met Leu Leu Leu Phe Leu Pro His Pro Ser Pro Pro Leu Leu Pro Pro
1 5 10 15
Ala Ala Ser Arg Thr Arg Pro Pro Pro Arg Leu Leu Leu Pro Pro Ile
20 25 30
His Ala Ser Pro Ser Pro Glu Leu Leu Ala Lys Ser Ala Leu Arg Arg
35 40 45
Ile Ser Asp Lys Leu Arg Ser Leu Gly Tyr Leu Glu Ala Asp His Pro
50 55 60
Glu Ala Ala Pro Gly Pro Ala Ala Pro Glu Ala Gly Ala Gly Ala Ser
65 70 75 80
Pro Gly Glu Ile Phe Val Pro Thr Pro Ala Gln Leu Pro Arg His Arg
85 90 95
Val Gly Ser Thr Leu Asp Pro Ser Trp Ala Thr Gly Asp Gly Glu Gly
100 105 110
Ala Ala Ala Ser Arg Arg Arg Arg Arg Gly Gly Arg Asp Ser Ser Ala
115 120 125
Ala Ala Ser Ala Pro Pro Ser Ala Ala Glu Leu Ala Leu Pro Arg Asp
130 135 140
Glu Leu Arg Arg Leu Gln Gly Ala Gly Ile Arg Leu Arg Asn Arg Leu
145 150 155 160
Lys Val Gly Lys Ala Gly Val Thr Glu Gly Ile Val Asn Gly Ile His
165 170 175
Glu Arg Trp Arg Asn Ala Glu Leu Val Lys Ile Arg Cys Asp Asp Val
180 185 190
Ser Ala Met Asn Met Lys Arg Thr His Glu Ile Leu Glu Arg Lys Thr
195 200 205
Gly Gly Leu Val Ile Trp Arg Ser Gly Ser Thr Ile Ile Leu Tyr Arg
210 215 220
Gly Thr Asp Tyr Lys Tyr Pro Tyr Phe His Asp Arg Glu Met Lys Asn
225 230 235 240
Asp Met Asp Glu Ser Ser Glu His Thr Ser Ser Asp Asp Glu Asp Ala
245 250 255
Asp Leu Ala Ile Ile Ala Ser Glu Gln Ser Gly Ser Glu Glu Asp Ser
260 265 270
Asp Asn Pro Ala Glu His Gly Ser Asn His Thr Glu Glu Gly Asp Asp
275 280 285
Leu Thr Arg Arg Phe Gly Val Asp Ala Leu Glu Gly Asn Leu Asp Ile
290 295 300
Gly Ser Ala Glu Gln Ser Ile Asn Ser Ala Thr Lys Asp Gln Gln Ala
305 310 315 320
Ile Leu His Thr Ser Thr Asn Val Ser Arg Pro Ser Glu Ile Ser Gly
325 330 335
Arg Ala Arg Ser Thr Leu Val Ala Gly Val Gly Ser Pro Asn Lys Phe
340 345 350
Arg Leu Gln Leu Pro Gly Glu Val Lys Leu Ala Glu Glu Ala Asp Lys
355 360 365
Leu Leu Asp Gly Leu Gly Pro Arg Phe Ser Asp Trp Trp Gly Tyr Asp
370 375 380
Pro Leu Pro Val Asp Ala Asp Leu Leu Pro Ala Ile Val Pro Gly Tyr
385 390 395 400
Arg Arg Ser Ser Asn Leu Gln Gly Leu Ala Ala Ser Met Ile Lys Leu
405 410 415
Trp Glu Arg Cys Glu Val Ala Lys Val Ala Ile Lys Arg Gly Ala Glu
420 425 430
Asn Ile Asp Ser Asp Leu Ile Ser Glu Lys Leu Lys Gly Leu Thr Gly
435 440 445
Gly Thr Leu Leu Ser Arg Asp Asn Glu Ser Ile Val Phe Tyr Arg Gly
450 455 460
Lys Asp Phe Leu Pro Thr Ala Val Ser Leu Ala Ile Glu Lys Arg Arg
465 470 475 480
Lys Tyr Gly Asn Ser Thr Ile Ser Asn Pro Lys Leu Asn Phe Asp Lys
485 490 495
Ser Thr Pro Gln Asn Ser Ser Lys Leu Lys Met Ala Thr Asp Val Ser
500 505 510
Leu Asp Gly His Glu Cys Tyr Glu Lys Lys His Lys Asp Glu Thr Ala
515 520 525
Val Ser Asp Asn Arg Ala Glu Ser Leu Asn Val Phe Ala Gln Asn Val
530 535 540
Glu Ala Arg Leu Ser Gln Ala Ile Ala Glu Lys Glu Lys Thr Glu Lys
545 550 555 560
Leu Ile Glu Glu Leu Glu Met Ser Ser Glu Pro Ser Arg Ala Glu Thr
565 570 575
Arg Glu Val Ile Ser Glu Glu Glu Arg Tyr Met Leu Arg Lys Val Gly
580 585 590
Leu Lys Met Lys Ser Phe Leu Leu Leu Gly Arg Arg Gly Val Phe Asp
595 600 605
Gly Thr Val Glu Asn Met His Leu His Trp Lys Tyr Arg Glu Leu Val
610 615 620
Lys Ile Ile Cys Lys Glu His Asn Ile Lys Asp Val Glu Tyr Ala Ala
625 630 635 640
Arg Thr Leu Glu Ala Glu Ser Gly Gly Ile Leu Val Ala Val Glu Arg
645 650 655
Val Ser Lys Ala His Ala Ile Ile Ile Tyr Arg Gly Lys Asn Tyr Gln
660 665 670
Arg Pro Ser Thr Leu Arg Pro Lys Ser Leu Leu Asn Lys Lys Asp Ala
675 680 685
Leu Lys Arg Ser Val Glu Tyr Gln Arg Tyr Lys Ser Leu Lys Leu His
690 695 700
Val Leu Asn Leu Ser Lys Asn Ile Asp Tyr Leu Lys Asp Gln Met Phe
705 710 715 720
Phe Lys Gln Met Glu Val Gln Pro Val Thr Pro Thr Asn Gly Met Asn
725 730 735
Ser Gly His His Asn Gln Gly Ile Leu Asp Leu Asn Val Asn Ser Gly
740 745 750
Thr Leu Val Asp Lys Lys Glu Glu Val Ser Glu Val Leu Pro Glu Cys
755 760 765
Ala Lys Ser Val Val Val Glu Cys Ser Ser Gly Glu Ser Glu Thr Glu
770 775 780
Gly Thr Ser Val Leu Thr Lys Ser Gly Val Pro Leu Asp Val Met Gln
785 790 795 800
Asn Lys Leu Leu Cys Phe Ser Lys His Thr Asp Asp Leu Ser Glu Thr
805 810 815
Thr Ser Ser Cys Leu Thr Glu Ser Thr Ser Thr Ser Ser Glu Ser Thr
820 825 830
His Gln Ser Pro Leu Ser Ser Ser Val Met His Asn Ser Asp Ser His
835 840 845
Arg Val Ser Gly Ser Lys Phe Val Gly Thr Leu Thr Pro Val His Glu
850 855 860
Leu Lys Leu Asp Glu Lys Ser Ser Gln Leu Pro Ser Ala Ala Ala Pro
865 870 875 880
Leu Ser Asn Arg Glu Arg Leu Met Leu Arg Lys Gln Ala Leu Lys Met
885 890 895
Lys Lys Arg Pro Val Leu Ala Val Gly Arg Asn Asn Val Ile Thr Gly
900 905 910
Val Ala Lys Ala Ile Lys Thr His Phe Lys Lys His Pro Leu Ala Ile
915 920 925
Val Asn Ile Lys Asn Arg Ala Asp Gly Thr Pro Ile Gln Gln Leu Ile
930 935 940
Ser Glu Leu Glu Glu Ala Thr Gly Ser Val Leu Val Ser Arg Glu Pro
945 950 955 960
Asn Lys Val Ile Leu Tyr Arg Gly Trp Gly Ala Asp Val Ala Gln Asn
965 970 975
Ser Leu Ser Gly Asn Asn Ser Thr Glu Gln Val Glu Lys Glu Val Ile
980 985 990
Ser Pro Gln Leu Leu Glu Ala Val Arg Leu Glu Cys Gly Leu His Pro
995 1000 1005
Gly Glu Ser Glu
1010
Claims (4)
1. a rice seedling yellow leaf gene, it is characterised in that: its nucleotide sequence is as shown in SEQIDNo.1.
2. a pair is used to detect the specific dCAPSPCR primer of a rice seedling yellow leaf gene described in claim 1,
It is characterized in that, nucleotide sequence are as follows:
Upstream: 5 '-GTGTGTACTCTCAGATGAAGTACTTGGACTCTCAGT-3 '
Downstream: 5 '-CACATTGTATTTGCAGAACA-3 '
That is sequence shown in SEQIDNo.3 and SEQIDNo.4.
3. a kind of method for detecting rice seedling yellow leaf gene described in claim 1, which is characterized in that use claim
It specific dCAPSPCR primer pair common rice described in 2 and is carried out containing rice yellow leaf trans-genetic hybrid rice seedling stage extracted DNA
Pcr amplification reaction, obtain 115bp nucleotide sequence as shown by seqid no.5 with 113bp nucleotide sequence such as SEQIDNo.6 institute
Show.
4. the detection method of rice seedling yellow leaf gene as claimed in claim 3, which is characterized in that by common rice " Jiahua-1
And OryzasativaLcv.Nipponbare " the obtained 115bp segment of DNA cloning, can by Eam1105I digestion at 83bp and two sections of 32bp, and by containing Huang
It is mono- section of 113bp that the amplification of leaf trans-genetic hybrid rice, which still keeps length,.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103923890A (en) * | 2014-05-05 | 2014-07-16 | 扬州大学 | Rice leaf color controlling gene LYL1 and application thereof |
| CN104404061A (en) * | 2014-12-03 | 2015-03-11 | 西南大学 | Yellow green leaf mutant gene YGL6 of rice, protein encoded by yellow green leaf mutant gene YGL6 and application of yellow green leaf mutant gene YGL6 |
| CN105420256A (en) * | 2015-12-31 | 2016-03-23 | 西南大学 | Rice yellow-green leaf mutation gene YGL8, protein coded by rice yellow-green leaf mutation gene YGL8, and application of rice yellow-green leaf mutation gene YGL8 |
| CN105713910A (en) * | 2016-01-28 | 2016-06-29 | 上海师范大学 | Rice leaf color gene regulated and controlled by temperature and detecting method and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103923890A (en) * | 2014-05-05 | 2014-07-16 | 扬州大学 | Rice leaf color controlling gene LYL1 and application thereof |
| CN104404061A (en) * | 2014-12-03 | 2015-03-11 | 西南大学 | Yellow green leaf mutant gene YGL6 of rice, protein encoded by yellow green leaf mutant gene YGL6 and application of yellow green leaf mutant gene YGL6 |
| CN105420256A (en) * | 2015-12-31 | 2016-03-23 | 西南大学 | Rice yellow-green leaf mutation gene YGL8, protein coded by rice yellow-green leaf mutation gene YGL8, and application of rice yellow-green leaf mutation gene YGL8 |
| CN105713910A (en) * | 2016-01-28 | 2016-06-29 | 上海师范大学 | Rice leaf color gene regulated and controlled by temperature and detecting method and application thereof |
Non-Patent Citations (2)
| Title |
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| 一个新的水稻叶绿素缺失黄叶突变体的特征及基因分子定位;刘朝辉 等;《遗传》;20120229;第34卷(第2期);223-229 |
| 一个新的水稻幼苗黄叶突变体的遗传分析及其基因的初步定位;葛绍兴 等;《上海师范大学学报(自然科学版)》;20130228;第42卷(第1期);49-54 |
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