CN1923850A - Control gene of paddy leaf shape and application thereof - Google Patents
Control gene of paddy leaf shape and application thereof Download PDFInfo
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- CN1923850A CN1923850A CNA2006100528408A CN200610052840A CN1923850A CN 1923850 A CN1923850 A CN 1923850A CN A2006100528408 A CNA2006100528408 A CN A2006100528408A CN 200610052840 A CN200610052840 A CN 200610052840A CN 1923850 A CN1923850 A CN 1923850A
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Abstract
the invention discloses a rice leaf shape controlled protein coded by SLL1 gene with amino acid sequence as follow as Seq ID No:2 as well as nucleic acid sequence in the Seq ID No:1 and plasmid, plant express carrier and host cell with the related gene. The invention also provides culturing method of coiled plant leaf, which transforms plant carrier into plant cell to form plant. The new gene SLL1 is cloned from coiled rice leaf mutant to code transcription factor of MYB-DNA connecting area of SHAQKYF, which controls coiling degree through moduling the programming death in the vascular tissue differentiation course.
Description
Technical field
The invention belongs to plant genetic engineering field.Specifically, the present invention relates to a kind of map based cloning technology of utilizing and clone paddy rice SLL1 (Shallot Like Leaf 1) gene, and utilize transgene complementation test to identify the function of this gene; Also relate to simultaneously and utilize this gene regulating blade thick walled fiber tissue, thereby, increase mesophyll cell number and the chlorophyll content of farm crop by regulating the blade interior structure of paddy rice; Utilize this generegulation leaf rolling simultaneously,, improve the output of farm crop in order to obtain the ideotype of farm crop.
Background technology
Paddy rice has the typical feature of monocotyledons leaf as monocotyledonous model plant.The paddy rice leaf is made up of blade and leaf sheath, and blade can be divided into epidermal area, mesophyll layer and three integral part of dimension pipe layer.In dimension pipe layer, comprise the phloem near abaxial side, xylem, the vascular bundle sheath parenchyma cell of periphery and the thick walled fiber cell that vascular bundle and epidermis are closely linked to each other of close adaxial and its surface.In normal blade, sclerenchymatous cell appears at adaxial and its surface and abaxial side, plays mechanical support.Sclerenchymatous cell is this living wall and the lignified dead cell with uniform thickened, and the sclerenchyma in the vein mainly includes the tracheid of transportcapacity and the thick walled fiber cell of epidermis below.Be differentiated to form lignifying conduit element from mesophyll cell, the differentiation of dead cells such as sclerenchyma is a typical example of programmed cell death.It is the classical system of research dimension pipe differentiation that people utilize Herba Zinnia elegansae (Zinnia) mesophyll cell of vitro culture.And the research in paddy rice does not appear in the newspapers as yet.
Leaf development is a very complicated process, has comprised all many-sides such as cell fission and extension, polarity decision, tissue differentiation.Paddy rice is one of most important food crop in the world, and the research of leaf morphology receives much concern always.Up to now, in paddy rice, find nearly 11 leaf roll locus, only for aspects such as what phenotype, physiology and the first location of heredity, do not seen the report of successfully cloning the rice leaf roll related gene so far yet at present.
The present invention utilizes the leaf extreme mutant that curls, at first be cloned into the SLL gene by the map based cloning technology, a kind of MYB-DNA of this genes encoding is in conjunction with the transcription factor in territory, and, control leaf morphology by influencing sclerenchymatous formation with this programmed death process of controlling in the rice leaf dimension pipe differentiation in the mesophyll cell.The blank that the programmed cell death problem of heavy wall element is still studied in the rice leaf, but former studies show that, programmed cell death and plant stress-resistance are closely related, and existing report is also thought Myb transcription factor and degeneration-resistant close contact arranged.
The research of leaf deformity in Arabidopis thaliana and corn mainly concentrates on aspect polarity builds up, the foundation of blade adaxial and its surface attribute needs PHAN family and HD-ZIP III family etc., the foundation of blade abaxial side attribute needs YABBY family and KANADI family etc., and these property decision genes majorities are encoded and transcribed relevant regulatory factor.
Summary of the invention
The technical problem to be solved in the present invention provides a kind of new gene SLL1 that clones from the rice leaf roll mutant, the MYB-DNA of a SHAQKYF class of this genes encoding mainly controls leaf amount of crimp by the programmed cell death in the differentiation of regulation and control abaxial side vascular tissue in conjunction with the transcription factor in territory.
In order to solve the problems of the technologies described above, the invention provides a kind of control gene of paddy leaf shape encoded protein matter, this protein has the aminoacid sequence shown in the Seq ID No:2.
Improvement as control gene of paddy leaf shape encoded protein matter of the present invention: aminoacid sequence also is included in aminoacid sequence or the derivative that add, replace, insert or delete the homologous sequence of one or more amino acid or other species in the aminoacid sequence shown in the SeqID No:2 and generate.
The present invention also provides coding above-mentioned proteinic gene, and this gene has the nucleotide sequence shown in the Seq ID No:1.
Improvement as gene of the present invention: nucleotide sequence also is included in the nucleotide sequence shown in the Seq ID No:1 and adds, to replace, and inserts or lack one or more Nucleotide and the mutant, allelotrope or the derivative that generate.
The present invention also provides the plasmid that contains said gene.
The present invention also provides the plant expression vector that contains said gene.
The present invention also provides a kind of host cell, and this host cell contains gene order.
Improvement as host cell of the present invention: this cell is Bacillus coli cells, agrobatcerium cell or vegetable cell.
The present invention also provides a kind of method of the leaf rolling that cultivates plants, and comprises with above-mentioned plant expression vector transformed plant cells, again the plant transformed cell culture is become plant.
Improvement as method of cultivation of the present invention: transform and adopt agrobacterium-mediated transformation or particle bombardment.
Specifically: the new gene SLL1 that clones from the rice leaf roll mutant provided by the present invention has the dna sequence dna shown in Fig. 4 and seq ID No:1.Be also included within the mutant allele that replaces a Nucleotide and produce, also contain the gene order that has identical function and can reach the object of the invention.
The protein shown in the Seq ID No:2 belongs to the transcription factor protein family of the MYB-DNA of what SHAQKYF class in conjunction with the territory among the present invention, wherein carries out one or several amino acid whose replacement, insertion or the disappearance functional analogue that amino acid obtained.SLL1 and Arabidopis thaliana KANADI albumen homology are 56%; The transcription factor of a kind of GARP by name of KANADI encoding histone family also have MYB-DNA in conjunction with the territory, and the polarity of reconciliation statement fur and carpel is grown.
Gene or the segmental carrier of portion gene that contains sequence shown in Fig. 4 and the seq ID No:1 provided by the present invention, as shown in Figure 7, this carrier can be expressed by the polypeptide of above-mentioned nucleic acid sequence encoding or homology analogue.
The method of cultivation that plant leaf provided by the present invention curls is a kind of method of carrying out the high-efficiency plant genetic transformation with SLL1; Specifically, be to utilize the plant expression vector transformed plant cells to influence farm crop leaf morphology method.
Realize that concrete technological step of the present invention is as follows:
One, the separation of rice leaf roll mutant SLL1 and genetic analysis:
The extremely curling leaf roll mutant body of rice leaf of the present invention is with chemical mutagen (the ethyl methane sulphonate of " Japan is fine " japonica rice variety (japonica) through 1% concentration, EMS) after the processing, the mutant sll-1 that the phenotype that obtains by a large amount of screenings is stable again.This mutant except blade table reveals curl, other phenotype is all similar to wild-type, as shown in Figure 1.A large amount of hybrid experiments prove that we are resulting to be a cryptic mutant that meets single-gene control of heredity rule.
Two, the leaf SLL1 gene of map based cloning control paddy rice:
1, the Primary Location of SLL1 gene:
In order to separate the SLL1 gene, the present invention has at first set up a target group that big polymorphism is high, the F2 colony that forms by No. 6 (Indica) hybridization of sll-1 and rice variety Nanjing, method by map based cloning again, and utilize STS, SSR equimolecular mark that Primary Location is carried out in the SLL1 site, with its Primary Location the 9th chromosomal long-armed on, and between RM3700 and RM1896 two SSR marks, see Fig. 2.
2, the Fine Mapping of SLL1 gene:
Sequential analysis by to BAC clone B1040D06 has developed 2 new SSR marks and 12 STS marks, and is as shown in table 1, SLL1 accurately is positioned between STS mark T5904-7 and the T5904-9, within the scope of 29.57kb.Open reading frame (ORF) by this section of sequencing analysis is inferred candidate gene, the SLL1 gene accurately is positioned the position (Fig. 3) of the 103061-108515 of BAC clone B1040D06.
Table 1, new development have polymorphic SSR mark and a STS mark
The primer title | Clip size (bp) | Preceding primer (5 '-3 ') | Reverse(5’-3’) |
T5904- 4 | 246 | CAGGTGGTGACCATTCTAAA | CAGAGTCCTTGATTGACCTAAC |
S5904- 3 | 107 | AAGCACTCTCACACACAAACA | AGACAAGCAGCAAAGAAACAC |
T5904- 9 | 163 | CGTCAGCCTACACATCATCTC | CTTGAGATTGGTTTGCATCAT |
T5904- 10 | 195 | TGAACCAATTCTACCATCCAA | TCTCCCGTTGTGATTAAAGTG |
T5904- 1 | 219 | TGGATGAACTCCCATATCTCA | AAGGGTGACAAAACAGACACA |
S5904- 1 | 118 | TGTTACAAACAAAACAAGCAAA | GTTGCTGCTCCTACCTCACT |
T5904- 5 | 204 | GACTATTGTGGGTTTCCCAGT | ACGTGCAACAACAATAAAAGG |
T5904- 7 | 181 | GTTGTTCACGACTCATCCATC | GAGACCAGTAACACCATCCAA |
T5556- 1 | 345 | CTGCACGATTGTATTTGGAAG | CTCCGCTTCAGTTTTACAATG |
T5577- 1 | 106 | CATATTATGGGACGGAGGAGT | TCGTGAGTCATGACAGTATGC |
T5421- 2 | 224 | TTCTAGCTTTGGGTACGACACG | ACAGCAAGCGAACGTACATATTC |
T6453- 3 | 265 | GGTTTCCCAGTAGTAGTGAC | CTGGCTCAATGATTACAAG |
T5707- 1 | 189 | CCTAACGAGCTAAAAACCCC | CATGGAGGAGGAAAGCGAC |
T5861- 2 | 173 | TGGTTCCTCATTTTACATTGC | TGGGCAAACCTGTCTAAAAC |
3, the evaluation of SLL1 gene and functional analysis:
Utilize the complementary carrier (Fig. 7) of pCAMBIA1300 plasmid construction, pass through transgenic technology, the transgenic research that has complementary functions, the result shows that the present invention has obtained to make mutant to recover the transgenic paddy rice (Fig. 8) of normal function, proved that the present invention has correctly cloned the SLL1 gene, dna sequence dna of clear and definite SLL1 gene (Fig. 4) and cDNA sequence (Fig. 5), amino acid sequence analysis show the transcription factor (Fig. 6) of the MYB-DNA of a SHAQKYF class of SLL1 genes encoding in conjunction with the territory.CDNA sequence shown in Figure 5, its nucleotide sequence are Seq ID No:3.
In paddy rice, the forfeiture of SLL1 gene function causes the disappearance of blade abaxial side thick walled fiber cell, and causes blade curling to adaxial and its surface.Further experiment shows, the MYB-DNA of a SHAQKYF class of this genes encoding mainly controls leaf amount of crimp by the programmed cell death in the differentiation of regulation and control abaxial side vascular tissue in conjunction with the transcription factor in territory.This gene is disclosing the molecule mechanism that leaf generation is curled, and understands the differentiation and development of blade mesophyll cell, vascular tissue and thick walled fiber tissue, and monocotyledons blade polarity the aspect such as is built up and had important theory value.Leaf is the Main Agronomic Characters of crop, occupies an important position in breed improvement and the breeding of desirable strain shape molecular designing.Utilize transgenic technology can the leaf rolling degree of plant be improved, improve the chlorophyll content of blade and improve photosynthetic efficiency, quicken the accumulation of dry-matter, boost agricultural yield, have broad application prospects.
China be Rice Production and consumption big country, along with Increase of population, to the paddy rice demand with in rising trend.High yield and super high-yielding are the crop breeding Eternal Theme, are to guarantee China's grain security and the most effective guarantee of agricultural sustainable development.The clone of SLL1 gene and application have been established the molecular theory basis and the beneficial gene resource that can directly utilize are provided the molecular designing breeding of the desirable strain shape of super rice.No matter be that rice is done the reach of science, realize the breakthrough for the third time of China's rice yield, still, all be significant solving new millennium China's grain security problem.
Description of drawings
Fig. 1 is the blade phenotypic map of paddy rice fine wild-type of Japan and leaf roll mutant body sll1;
Fig. 2 is the Primary Location figure of SLL1 on paddy rice the 9th karyomit(e);
Fig. 3 is the Fine Mapping figure of SLL1 gene;
Fig. 4 is the dna sequence dna of 5LL1 gene;
Fig. 5 is the cDNA sequence of SLL1 gene;
Fig. 6 is the aminoacid sequence of SLL1 genes encoding;
Fig. 7 is complementary vector construction plasmid map;
Fig. 8 is the phenotypic map of function complementation experiment T1 for transgenic paddy rice.
Embodiment
With reference to above-mentioned accompanying drawing, the specific embodiment of the present invention is elaborated.
" a " among Fig. 1 represents leaf cross-section, and " b " represents the plant phenotype.
" sll1 " among Fig. 8 represents the leaf roll mutant body, and " sll1/SLL1 " representative is changeed SLL1 gene T1 for the mutant plant.
Embodiment 1:
1, rice material
Paddy rice (Oryza sativa L.) mutant Shallot-like leaf1 (sll1), original wild-type material are " Japan is fine " japonica rice variety.
2, analysis and target group
Hybridize for No. 6 in sll1 mutant that isozygotys and original wild-type kind Nanjing, and F1 obtains F2 colony for selfing, and therefrom select 711 sll1 mutated individuals as target group.Every strain of early tillering stage get 1 the gram about tender leaf, be used for extracting total DNA.
3, SSR and STS mark location SLL1 gene
Adopt the rapid extracting method of paddy rice minim DNA from rice leaf, to extract the genomic dna that is used for the assignment of genes gene mapping.Get about 100mg rice leaf, through liquid nitrogen freezing, pulverize in the little mortar of diameter 5cm is transferred in the 1.5ml centrifuge tube and is extracted DNA, and the DNA resolution of precipitate of acquisition is in 100 μ l ultrapure waters.Each SSR and STS react with 2 μ lDNA samples.
In the Primary Location stage of SLL1 gene, analyze carrying out SLLP by the individual microcommunity of forming of 152 F2, molecular genetic linkage map according to japonica rice of announcing and long-grained nonglutinous rice establishment, choose the approximate SLLP primer that is uniformly distributed on each bar karyomit(e), carry out pcr amplification according to known reaction conditions, separate and bromination second pyridine (EB) dyeing through 5% agarose gel electrophoresis, the polymorphism of detection PCR product, with the SLL1 Primary Location between No. 9 long-armed RM3700 of karyomit(e) and RM1896 mark.
4, the Fine Mapping of SLL1 gene
When Fine Mapping SLL1 gene, the sequential analysis by to BAC clone B1040D06 has developed two new SSR marks and 12 STS marks, and 711 mutated individuals consistent with mutation type surface selecting in the F2 colony are carried out SSLP and STS analysis.SLL1 accurately is positioned between STS mark T5904-7 and the T5904-9, within the scope of 29.57kb.
According to BAC clone B1040D06 sequence, design 40 pairs of primers, adopt PCR method from the Japanese fine genome of sll1 and wild-type, to amplify this 29.75kb respectively, carry out sequencing analysis.The product of finding mutant sll1 amplification is than the Japanese fine replacement (G is to A) that a base is arranged of wild-type.Finding that according to the cDNA level detection this catastrophic event occurs on the splice site, cause normally montage of mRNA.According to the gene annotation information (TIGR) of BAC clone B1040D06 sequence, MYB-DNA of this genes encoding is in conjunction with the transcription factor in territory.Open reading frame (ORF) by this section of sequencing analysis is inferred candidate gene, with the position of the SLL1 assignment of genes gene mapping in the 103061-108515 of BAC clone B1040D06.This gene comprises 6 exons (Exon) and 5 introns (Intron) altogether.
5, the prediction of the acquisition of SLL1 cDNA full-length gene and function
By the total length order-checking, pass through several cDNA full length sequences (Fig. 6) that amplification has obtained the SLL1 gene of taking turns.On the basis of above-mentioned research, found the position that gene is undergone mutation, and predicted the transcription factor of the MYB-DNA of a SHAQKYF class of this genes encoding, by the sclerenchymatous programmed cell death of regulation and control abaxial side, with the shape of regulation and control plant leaf in conjunction with the territory.
Embodiment 2:
Plant Transformation:
BAC is cloned B1040D06 carry out complete degestion with BglII and SSe8387I, extracting dna fragmentation about 8KB after the electrophoretic separation is connected on the isocaudarner BamHI and PstI point of contact in the pCAMBIA1300 multiple clone site, this dna fragmentation has covered the genome area of whole ORF, also comprises ATG upstream 1.6KB promoter sequence.
It is rice transformation among the EHA105 that this plasmid changes Agrobacterium (Agrobacterium tumefaciens) strain over to by the method that shocks by electricity.We utilize mutant rataria inductive callus, after the process inducing culture cultivated for 3 weeks, select the vigorous callus of growth as the acceptor that transforms.Infect rice callus with the EHA105 bacterial strain that contains the double base plasmid vector, under dark, 25 ℃ of conditions, cultivate 3 days altogether after, cultivate containing on the screening culture medium of 40mg/LHygromycin.The screening kanamycin-resistant callus tissue breaks up in advance on the substratum and to cultivate about 10 days containing 50mg/L.The callus of differentiation is in advance gone on the division culture medium and cultivates under illumination condition.Obtain the resistant transgenic plant about one month.Plant is identified and successive is observed, find that plant leaf has recovered normal form, in the mutant of same growth phase relatively, vane extension is curling, and to its discoverys of cutting into slices, the thick walled fiber cells whose development is recovered normally (Fig. 8).
At last, it is also to be noted that what more than enumerate only is several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.
Sequence table
SEQ ID NO:1
1 ATGGCGCCGA TGATGCTGCA GCCGCTCGAC GCCGGCGGCG GCGCGTCGGC GCCGCCGCCG
61 CCGATCCGCG GGATACCTAT CTACAACGGC CCCGGCGGGT TCCCGTTCCT GCAGCCGTCG
121 CCCACCGCCG GCGACGTCGG CCACCACCAC CACCACCACC CCAAGATGGG ATTCTACAGC
181 TCGTACCACC ACCCATCCAC GTGGCCCTCC ACGTCGCCGT CCCCGCTCGC GGCGCCGCCG
241 GGCGCCGCGT CGTCGCCGCT CGACCCCACG GCGGCGTTCC TCTCCTCCCC CCACCACCGG
301 ATGCTGTCCG CCGCCTCGGG GAGGCTCAAC GGCATGCTCT CCGTCTCCGA CACCCTCCGC
361 AGCTACGGCG TCCCCGGCGC CGCCGCCCCC GGCGTCATCG GCGGCGCGCA CCACCACCAC
421 CACCACCTCC ACGGCGGCCA GCCGTTCGTC GGCGCCCTCG CGTCCCGCTT CATGCCCAAG
481 CTCCCCGCCA AGCGCAGCAT GCGCGCGCCG CGCATGCGCT GGACGAGCAC CCTCCACGCC
541 CGCTTCGTCC ACGCCGTCGA GCTCCTCGGC GGCCACGAGA GTACGCCCCC GCCATCATCG
601 TCGTCGTCCT CCTCCTCCTT CTCACCACCA TCATCAACGA GCTCGATCAA TCTGATGAGA
661 ATCCAATCTT GTTCTTGGCA GGGGCGACGC CCAAGTCGGT GCTGGAGCTC ATGGACGTCA
721 AGGATCTGAC GCTAGCGCAT GTCAAGAGCC ACCTCCAGGT ATCCCGCCAT TGCCGACCAA
781 TTCTCACAGC TCGATCGATC GATCAAACAC CACTCACCAC TGCACCATCT CTCTCTTTCT
841 CTCTCTCGAT TTCGATTCCA TTCAGCTTCT GTTCTTGATC ATCTTGTTTG GGTGAGACAA
901 AGATGCCAAT GCAGTAACTC CTGAGTTAGT GAAGAAACTG CCATTCTTGG GAGCAGAGAG
961 AGAGAGTGTG TGTGTGACAT GTTTGGGGAT GTGTGTGCAA GAGAGAAGAG AGAGAGAGAG
1021 GGGAAAAAAG TTGCCATCAT TACACAATAA TGCATTGCAT CTGCATCTGT GCTACTAGCT
1081 CCTTCCTTAG CTCTAGTCAT ACTAGCGTAT ATGTGCACGG CCCAAAGCAC TCTCACACAC
1141 AAACACAAGA GAGAGAGAGA GAGAGAGAAG AAGAAGATGA GAGATGAGAG AGAGTAGGAC
1201 ATATTTTGTG TGTGTTTCTT TGCTGCTTGT CTTTTGCAAT GGCTTCAACC TCCTGGTTTC
1261 ATTTTGATCT CACAAAAAAT GTGTGTGTGT GTGTAAATAT ATATATTGAT CTTCTTTTTG
1321 TTGGACTTTT GTGTTTTATT ACTGCAGATG TATCGCACCG TGAAGAGCAC TGACAAGCCT
1381 GCAGCCTCTT CAGGTGATTT TTTTTACCAT GGCAGCTACT AGTAGTAGTA GTAGTAGCTA
1441 ATTTAGCTAA GCTCCATTTT GCTTTTCTTT TTTTTTGGCG TTGTGCATTG CGTTTTACAT
1501 TACATTGTTG CTATGGTTTT TTTGACGTGT AGGGCCGGCG GACGGCGGCT CCGGCGACGA
1561 GGAGTTCGCC GGCGGCGGGC AGGCGGCGTC GGGCGGCGGC GACAGCATGT GCCTGAGGGG
1621 TGGCGGCGGC GGCGGGGTGG CCGCGGCGGC GTTCGCGGAG CACGGCCGGT CGGCGTCGGA
1681 GGGCGCCGCC AGCTCGGTCG GCGGCGGCGG CGGCGGCGAC ATGGACCAGT CGTCGGCCGG
1741 CAACACCAGC ACCACCAGGT GGAGCAACTC CTCAAGGTAC ACACATGGCA GAGTTGCATT
1801 TCAAGTCTTT CAGATTAATT ATATATCTAC ATTGTTATGC CATCTTTAAT TAATTAAAAA
1861 TGAAATTAAT CAATCAGAAT GTCATGAGAT GATGATGATG ATGATAATAA TACTAGGATA
1921 GGAAGAGGAT GATATGATGG ATGGGTGTGC TATGTGTGTT TTTAAGTGTG TTTTAGCTTA
1981 GAACCAAAGG CAACACCAAC AAACTTTGGC ATGCATTGAT GATGTATCTA AATATGAGTT
2041 AGTGACATGC AAAATTAACC TGCCAAGTAC AACCTCTATT TATTAATTCC CAAAAAAATC
2101 TCATGGAGGA GAAGGGAGAG AGAGAGATAG TAGTAATGTG TGGTGATGCA TGGCTAAAAA
2161 AGCTTAGCAA AAGAATGTTT TGGGGGCACA CACACACACA CCTGGAGAGG GAGTTGCCTA
2221 TGTTTAGGTG GCCAATGTTG TTATGTCACA CAAAAGACCA AAAAGTCACA CACCCCATGG
2281 CTTGGCCTCC TCCTCCCCCC TTGACCTGTG CAGTTAGCTA GGCTTCTTCT CTTCTCTCCC
2341 TCTGTGTTTG GTTTTTGTGG AGCTAAAAAC ATTTTAAAAA TCTTGCAAAC ATTGTGTGTA
2401 AAAATAGGTT CTTGGGCTCT GCCCATTGTC CTCTAGCTTC TCTCAAATCT CTCTCTCTCT
2461 CTCTCTCTCT CTCTCTCTCT TTCTCAAACC TAGGGAGGGG AGGGTTACTC AGGCAGGGCT
2521 CCATTATCAT TTCTTATCAA TGCATGGCTC TGATTCCAGG GGGATGGGGC TTTGACCTGA
2581 CATGCCCTAT TGCCTAGGAA TATGTAACCA AGGACTTGCT GCATTCTTGG TTCTTTCTCC
2641 CTCTCTCACC AGTCTCTTCA ATGCCCTTTT CTTCCACCTC TTGGGCAATT ATTAGCATGG
2701 ATGGAGCTCA CCTTTTCACT CTACCCTCTA GCTGTGATTA AATCATTGAC TAATCCTAAT
2761 TCATAAATAT AAGGGATTCA ACTATGTAAC TAGATGTACT AGATCCTAGT ATTACTAGTG
2821 TGTTTTGATT CTGTTAGGAG GAGTGATTAG CGTTTGTCGT AAGTACTAGT ATTAAACTAT
2881 AGGGGATTAG TGTAGATCAT TTTTTTAAAA ATTAGTGAGT GTGCTTGGGC AGATTTTACT
2941 CCAGCCATTG GCGTGAAAAC TTTGATCTTT GGGCAGCTTG GAAGCAAGTA CAGTTCAACC
3001 TGTCCTTGGC TGCATTCGCT GCTTTTTCTT TCTCAGGACT CACTGCAACT TTCTTGCCAT
3061 CCCCAACACC TCCCTTTCGG TCGTGTTTGA TCAGTTGCAT TGCTGGTTTC TTCGCCGCTT
3121 TAATTTCTCC ATCTTTCTTC TGTTCCTTAT TGGCTAAGTA AAAAATCATT CTCAGCAATC
3181 AAGTTTTGAA ATTCAAATTA GCTTCAGGTT TTATCTATCT AATAATTCTC TGACTAGTTC
3241 TCTTTCTTTC AAGACCTGTT CTTGTTCAGA ACTCTTATGC AGTTGAGAGT TCAGTTGTTC
3301 ATGGAAGTAA TGGCAATGCC TGGTCTAGTC TTTGAGCAGT ATCCTCCTTA TGCTCTTGAT
3361 TTTCCTTCAC TTTAGCCGTT TCAGTCAAAC TTTGGGGACA AATTAAAGTC AGCTGGTCAA
3421 GTGCATACAT ACCACGTCAT ATTTTTTAGA TGAATCCATT GTAGTATTAA ATACTAGTGA
3481 CAAAAAAATA TAACTAAATG GCATTAAATT GCAACCTTCT TCCCCAAAAT AATTGGGCAC
3541 AACTACCTTG CTTGCATATG CTTTATAGTA CTCCTACTTA TTACTTCCTT GAATCCATGA
3601 GGATTCCAGG AATTTGGTCA GGTCAAATGT CCAGGAGGAA TTAGGCTATC TTTTTTTATC
3661 TTCTTCTTTT TCTTCTAGGT AGTAGGATCG TTGTGCACAT ATAAACAAAA CAAAATAGAA
3721 ATTTAAGAGG AAACCCTCCC AAAAAAAATA CGACTATCAA TTATGAGTGA TTTTACAGTT
3781 CTTGAGGAGG TACAATGATA CATTTTCTAT TTTAAAATTC AGTAACTTTG ATACTATGAG
3841 GTACAGAAAT TTGCACAAAA AAAATTTGGT ACCTCTCAAG TACTTAAAAT ATCTTATCAA
3901 TTATATTTCA GCAATGTTGT CGTTGCTCGA GTTGTGCAAA AACATTTCTG ATGAACAAGT
3961 GGAGATGTAT GCATATTTAG AGAGGGAAAC ACATGCATTT TTTAATATAC ATTCTCATGA
4021 CAAATTTTGG TGTTCTCGTC TCATTTTAGT CTAGCTTAGA TTAATGGCAT CCATATATTG
4081 GTGAACTGAA ACAGATTGTA TATGTCCAGT CTTGAACAAA CTTCATTATA CAATGCATAA
4141 TCAGAAAAGA TACAGACATA TATTGGCAGA TCACATAAAG TGGTGTATCT ATGATTGGGT
4201 GGTATTTTGG ACATACAAAG TCTACTGGTC CCTCATTTGT TAACAGCATA CATTTGTTCC
4261 TATCAAGACA AAAGCACTTT ACAACAGTTT GGTAGTAAGT TGGTGTTATG TGCGTGTCTA
4321 TTTAGATAGA GTGGTAGTAA AGGAAGGACA CTTTTTCTTG ATATACCAAA ATCACACATA
4381 AAAAAGAAGT GTCATTTTCA TGATCAACAT GTGGGGTATG AACAAAAATG TTCTCCCCCC
4441 TCCAACCAAC ATTGAAGTCA TCACACATAA AAAGCAAAAA CATTTAACAA TAATACATGC
4501 AGACACATGT GCATGCATGT GTGGATCACG CAAGGACATA CTAATTCAAT TAGGTATTCA
4561 CAATTAATTA GTGCAGGAAA TGACATGTAA TTACAATTAT ATATGCACAA CGATCCTTGT
4621 GTATTTGTGT GACCCATTGT AGTAGCATTT GATTCCTTTC TTGTGCCTTT CTCTACATTG
4681 AATTCGATCT TTGCCAACAG CGGCTTGCTG ATGAGCTCCT GTGCCATTGC ACTGCTGAGC
4741 TGCACCAAAC AGGGACATCT TTTAAGAGCT TGTTGCCTGG AAATGATCCA TGTAACCAAC
4801 TGTTCAGGAA GAGCACACTG TTTGCTGCTC ACTTTAGCTA AAGCAAAACT GTGTGCATTA
4861 TGCAATGCAT ATGCATCCTT CCATCTACCC CTGTGTCCTT TGCCCCCCAA ACTGTGTAAC
4921 TACAGTTTAC CATGCACATA CATACAATGC ACATCTACCC AATTGTAGCA TTGTTGCAAG
4981 GCCTACTTCA GAATTCAGTT AGTCCCCAAT TGATCCAACT GCACACATAT ACATTTACAT
5041 ATACATATAC ACAAACATAC CAGCTTGGAG AAAAGGTTGT AACAACTTTG CTAAATGTGA
5101 GATGTGTAAT GTGTGTTCTT CCTCTTTTCA GGGACCCATG GCTGTCGTCC AATTCTTGCA
5161 ACATGGACGC CCATCGCTCC GTAGGATTGT CTTCTCCTAT TGAGGTATAT ATCCTTACAT
5221 TGTTCAAAGC TTCTTTCGAT TCCTAGCTAA CTGCATATCC ATGCAGAAAT TCATTAGCTC
5281 GCATTCATTC TGAACATGAT GTCTGAAACT CTGAATTTTT TTTCGTTTGT GCGTGTTTCA
5341 GAACTTGGAA CCGTGCAGAT CGAGCAGCTC GCAGGTGTCC AACCATGAGC TGAGTAGCCC
5401 TAGTCTCGAG TTCACTCTAG GGAGGCCTGA CTGGCACGGT GCAGATCATG ATTAG
SEQ ID NO:2
Met Ala Pro Met Met Leu Gln Pro Leu Asp Ala Gly Gly Gly Ala Ser Ala Pro Pro Pro
1 5 10 15 20
Pro Ile Arg Gly Ile Pro Ile Tyr Asn Gly Pro Gly Gly Phe Pro Phe Leu Gln Pro Ser
21 25 30 35 40
Pro Thr Ala Gly Asp Val Gly His His His His His His Pro Lys Met Gly Phe Tyr Ser
41 45 50 55 60
Ser Tyr His His Pro Ser Thr Trp Pro Ser Thr Ser Pro Ser Pro Leu Ala Ala Pro Pro
61 65 70 75 80
Gly Ala Ala Ser Ser Pro Leu Asp Pro Thr Ala Ala Phe Leu Ser Ser Pro His His Arg
81 85 90 95 100
Met Leu Ser Ala Ala Ser Gly Arg Leu Asn Gly Met Leu Ser Val Ser Asp Thr Leu Arg
101 105 110 115 120
Ser Tyr Gly Val Pro Gly Ala Ala Ala Pro Gly Val Ile Gly Gly Ala His His His His
121 125 130 135 140
His His Leu His Gly Gly Gln Pro Phe Val Gly Ala Leu Ala Ser Arg Phe Met Pro Lys
141 145 150 155 160
Leu Pro Ala Lys Arg Ser Met Arg Ala Pro Arg Met Arg Trp Thr Ser Thr Leu His Ala
161 165 170 175 180
Arg Phe Val His Ala Val Glu Leu Leu Gly Gly His Glu Arg Ala Thr Pro Lys Ser Val
181 185 190 195 200
Leu Glu Leu Met Asp Val Lys Asp Leu Thr Leu Ala His Val Lys Ser His Leu Gln Met
201 205 210 215 220
Tyr Arg Thr Val Lys Ser Thr Asp Lys Pro Ala Ala Ser Ser Gly Pro Ala Asp Gly Gly
221 225 230 235 240
Ser Gly Asp Glu Glu Phe Ala Gly Gly Gly Gln Ala Ala Ser Gly Gly Gly Asp Ser Met
241 245 250 255 260
Cys Leu Arg Gly Gly Gly Gly Gly Gly Val Ala Ala Ala Ala Phe Ala Glu His Gly Arg
261 265 270 275 280
Ser Ala Ser Glu Gly Ala Ala Ser Ser Val Gly Gly Gly Gly Gly Gly Asp Met Asp Gln
281 285 290 295 300
Ser Ser Ala Gly Asn Thr Ser Thr Thr Arg Trp Ser Asn Ser Ser Arg Asp Pro Trp Leu
301 305 310 315 320
Ser Ser Asn Ser Cys Asn Met Asp Ala His Arg Ser Val G1y Leu Ser Ser Pro Ile Glu
321 325 330 335 340
Asn Leu G1u Pro Cys Arg Ser Ser Ser Ser Gln Val Ser Asn His Glu Leu Ser Ser Pro
341 345 350 355 360
Ser Leu Glu Phe Thr Leu Gly Arg Pro Asp Trp His Gly Ala Asp His Asp
361 365 370 375
SEQ ID NO:3
1 ATGGCGCCGA TGATGCTGCA GCCGCTCGAC GCCGGCGGCG GCGCGTCGGC GCCGCCGCCG
61 CCGATCCGCG GGATACCTAT CTACAACGGC CCCGGCGGGT TCCCGTTCCT GCAGCCGTCG
121 CCCACCGCCG GCGACGTCGG CCACCACCAC CACCACCACC CCAAGATGGG ATTCTACAGC
181 TCGTACCACC ACCCATCCAC GTGGCCCTCC ACGTCGCCGT CCCCGCTCGC GGCGCCGCCG
241 GGCGCCGCGT CGTCGCCGCT CGACCCCACG GCGGCGTTCC TCTCCTCCCC CCACCACCGG
301 ATGCTGTCCG CCGCCTCGGG GAGGCTCAAC GGCATGCTCT CCGTCTCCGA CACCCTCCGC
361 AGCTACGGCG TCCCCGGCGC CGCCGCCCCC GGCGTCATCG GCGGCGCGCA CCACCACCAC
421 CACCACCTCC ACGGCGGCCA GCCGTTCGTC GGCGCCCTCG CGTCCCGCTT CATGCCCAAG
481 CTCCCCGCCA AGCGCAGCAT GCGCGCGCCG CGCATGCGCT GGACGAGCAC CCTCCACGCC
541 CGCTTCGTCC ACGCCGTCGA GCTCCTCGGC GGCCACGAGA GGGCGACGCC CAAGTCGGTG
601 CTGGAGCTCA TGGACGTCAA GGATCTGACG CTAGCGCATG TCAAGAGCCA CCTCCAGATG
661 TATCGCACCG TGAAGAGCAC TGACAAGCCT GCAGCCTCTT CAGGGCCGGC GGACGGCGGC
721 TCCGGCGACG AGGAGTTCGC CGGCGGCGGG CAGGCGGCGT CGGGCGGCGG CGACAGCATG
781 TGCCTGAGGG GTGGCGGCGG CGGCGGGGTG GCCGCGGCGG CGTTCGCGGA GCACGGCCGG
841 TCGGCGTCGG AGGGCGCCGC CAGCTCGGTC GGCGGCGGCG GCGGCGGCGA CATGGACCAG
901 TCGTCGGCCG GCAACACCAG CACCACCAGG TGGAGCAACT CCTCAAGGGA CCCATGGCTG
961 TCGTCCAATT CTTGCAACAT GGACGCCCAT CGCTCCGTAG GATTGTCTTC TCCTATTGAG
1021 AACTTGGAAC CGTGCAGATC GAGCAGCTCG CAGGTGTCCA ACCATGAGCT GAGTAGCCCT
1081 AGTCTCGAGT TCACTCTAGG GAGGCCTGAC TGGCACGGTG CAGATCATGA TTAG
Claims (10)
1, a kind of control gene of paddy leaf shape encoded protein matter, it is characterized in that: this protein has the aminoacid sequence shown in the Seq ID No:2.
2, control gene of paddy leaf shape encoded protein matter according to claim 1 is characterized in that: described aminoacid sequence also is included in aminoacid sequence or the derivative that add, replace, insert or delete the homologous sequence of one or more amino acid or other species in the aminoacid sequence shown in the Seq ID No:2 and generate.
3, a kind of coding claim 1 or 2 described proteinic genes, it is characterized in that: this gene has the nucleotide sequence shown in the Seq ID No:1.
4, gene according to claim 3, it is characterized in that: described nucleotide sequence also is included in the nucleotide sequence shown in the Seq ID No:1 and adds, to replace, and inserts or lack one or more Nucleotide and the mutant, allelotrope or the derivative that generate.
5, a kind of plasmid that contains claim 3 or 4 described genes.
6, a kind of plant expression vector that contains claim 3 or 4 described genes.
7, a kind of host cell is characterized in that: this host cell contains claim 3 or 4 described gene orders.
8, host cell according to claim 7 is characterized in that: this cell is Bacillus coli cells, agrobatcerium cell or vegetable cell.
9, a kind of method of the leaf rolling that cultivates plants is characterized in that: comprise with the described plant expression vector transformed plant cells of claim 6, again the plant transformed cell culture is become plant.
10, method of cultivation according to claim 9 is characterized in that: agrobacterium-mediated transformation or particle bombardment are adopted in described conversion.
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CNA2006100528408A CN1923850A (en) | 2006-08-09 | 2006-08-09 | Control gene of paddy leaf shape and application thereof |
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CNA2006100528408A CN1923850A (en) | 2006-08-09 | 2006-08-09 | Control gene of paddy leaf shape and application thereof |
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Family
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Cited By (8)
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CN102219839A (en) * | 2011-04-25 | 2011-10-19 | 中国水稻研究所 | Gene SRL-1 for controlling leaf shape of paddy rice and application thereof |
CN102329805A (en) * | 2011-09-30 | 2012-01-25 | 复旦大学 | Coding sequence for OsMYB gene in rice and applications |
CN102676544A (en) * | 2012-05-25 | 2012-09-19 | 复旦大学 | Coding sequence of MYB family transcription factor gene OsMYB84 in rice and application of gene OsMYB84 |
CN101857633B (en) * | 2008-12-18 | 2012-10-03 | 中国水稻研究所 | Leaf-shaped control gene SRNL1 of rice and application thereof |
CN103172714A (en) * | 2011-12-21 | 2013-06-26 | 中国科学院遗传与发育生物学研究所 | Rice leaf rolling-associated protein OsMYB103L as well as encoding gene and application thereof |
CN105695428A (en) * | 2016-03-17 | 2016-06-22 | 武汉大学 | Rice cytoplasmic kinase gene OsBHL1 and encoding protein and application thereof |
CN107163113A (en) * | 2017-07-09 | 2017-09-15 | 中国水稻研究所 | The leaf roll of paddy rice half controls the purposes of gene SRL9 and its leaf improvement |
CN109456984A (en) * | 2018-12-30 | 2019-03-12 | 浙江师范大学 | Rice glume development gene AH1 and its application |
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GENBANK: "登录号AP005904", 《GENBANK》 * |
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Cited By (14)
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CN101857633B (en) * | 2008-12-18 | 2012-10-03 | 中国水稻研究所 | Leaf-shaped control gene SRNL1 of rice and application thereof |
CN102219839A (en) * | 2011-04-25 | 2011-10-19 | 中国水稻研究所 | Gene SRL-1 for controlling leaf shape of paddy rice and application thereof |
CN102219839B (en) * | 2011-04-25 | 2013-02-27 | 中国水稻研究所 | Gene SRL-1 for controlling leaf shape of paddy rice and application thereof |
CN102329805A (en) * | 2011-09-30 | 2012-01-25 | 复旦大学 | Coding sequence for OsMYB gene in rice and applications |
CN103172714B (en) * | 2011-12-21 | 2014-07-30 | 中国科学院遗传与发育生物学研究所 | Rice leaf rolling-associated protein OsMYB103L as well as encoding gene and application thereof |
CN103172714A (en) * | 2011-12-21 | 2013-06-26 | 中国科学院遗传与发育生物学研究所 | Rice leaf rolling-associated protein OsMYB103L as well as encoding gene and application thereof |
CN102676544B (en) * | 2012-05-25 | 2014-02-19 | 复旦大学 | Coding sequence of MYB family transcription factor gene OsMYB84 in rice and application of gene OsMYB84 |
CN102676544A (en) * | 2012-05-25 | 2012-09-19 | 复旦大学 | Coding sequence of MYB family transcription factor gene OsMYB84 in rice and application of gene OsMYB84 |
CN105695428A (en) * | 2016-03-17 | 2016-06-22 | 武汉大学 | Rice cytoplasmic kinase gene OsBHL1 and encoding protein and application thereof |
CN105695428B (en) * | 2016-03-17 | 2018-10-26 | 武汉大学 | Rice cytoplasmic kinase gene OsBH L1 and encoding protein and application thereof |
CN107163113A (en) * | 2017-07-09 | 2017-09-15 | 中国水稻研究所 | The leaf roll of paddy rice half controls the purposes of gene SRL9 and its leaf improvement |
CN107163113B (en) * | 2017-07-09 | 2020-11-24 | 中国水稻研究所 | Rice half-rolling leaf control gene SRL9 and application thereof in leaf shape improvement |
CN109456984A (en) * | 2018-12-30 | 2019-03-12 | 浙江师范大学 | Rice glume development gene AH1 and its application |
CN109456984B (en) * | 2018-12-30 | 2021-05-25 | 浙江师范大学 | Rice glume development gene AH1 and application thereof |
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