CN1477112A - Rice Tiller Control Gene MOC1 and Its Application - Google Patents
Rice Tiller Control Gene MOC1 and Its Application Download PDFInfo
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Abstract
The present invention utilizes rice single tiller mutant Mono culm 1(moc1) to clone and identify the gene capable of controlling rice tiller, and its name is MOC1. The transgenic function complementary assay shows that MOCI is the gene for controlling rice tiller, and the amino acid sequence comparison analysis shows that said gene belongs to the GRAS transcription factor family. The gene can be used for regulating tiller character of rice.
Description
Technical field
The invention belongs to plant genetic engineering field.Specifically, the present invention relates to a kind of map based cloning technology clone's MOC1 (MoNo CULML) gene, and utilize transgenic function complementation experiment to identify this gene; Also relate to contain this gene and its have the homogenic carrier of the homologous sequence of similar functions and other species and relate to utilize this gene or its functional analogue regulation and control plant tillering or thereby group structure that the side shoot generating ability is regulated plant improves output, quality and the nutrient optical energy utilization efficiency of farm crop.
Background technology
Tillering is the important morphological feature of most grasses.The every joint of the cane of gramineous crop all has bud, and bud can be grown the formation branch, and the branch base portion forms adventive root, and this class branch is tillered exactly.The morphological specificity and the stem of tillering are basic identical, tiller can separate with stem and survival separately.Tillering of gramineous crop has different types according to the difference of its happening part: the rhizome type is arranged, dredge tiller type and close tiller type.Rhizome type such as sugarcane, root stock be horizontal walk in ground, and distance is very long just to form upright seedling; Branch and the adventive root of dredging the tiller type are taken place by the subsurface stipes, and seedling is born in outside the leaf sheath, is inclined upwardly; The branch of close tiller type and adventive root are taken place by stipes near the ground, and seedling is born in the leaf sheath, and be parallel with maternal plant
(1)Rice tillering has close tiller type feature.Between paddy rice cane first segment meristem zone is arranged, the cell of this meristem zone divides and can extend between the envoy, regulates tillering node, makes the rudiment in the soil of the different transplanting degree of depth of tillering, and keeps the feature of close tiller type
(2)
Rice seedlings enter tillering phase when growing to 4 blades.Since most of vegetative organ of paddy rice as blade, tiller and root system all is to form in this period, so be the important period in the rice growth process tillering phase.Spike number is the important determinative of rice yield, and the individual plant tiller number is the important factor of decision spike number, and crossing low or too high tiller number all can influence output
(3)Therefore, ability for tillering is important economical character, is to influence the rice yield important factor.
Past mainly concentrates on morphologic observation and Crop Physiology aspect to the research of tillering.The physiological research of rice cropping has preferably the relation of tillering quantity and envrionment conditions illustrates, and comprises the adjusting that factors such as temperature, illumination, moisture, farming and mineral nutrition are grown to tillering, the influence to tillering of especially nitrogen nutrition level and temperature
(4)And the research of carrying out from genetic angle is later, and it is generally acknowledged tillers is subjected to polygene combined adjusting.Intergenic additivity effect is strengthened with plant strain growth the contribution of tillering, and plays the dominance effect, and non additivity effect and Effect of Environmental account for back burner between gene
(5)Other gene pairs tillering abilities are also influential as downgrade gene, have the plant of downgrading gene phenotype very strong tillering ability is arranged.Research for the molecule mechanism of tiller regulating is less, several mutant rcn1 of tillering less of the generation through gamma-ray and mutagenesis of Takamura and Kinoshita report is only arranged, 2,3,4,5 (rcn, reduced culm number).These mutant at high temperature can recover normal phenotype
(6)Moc1 (mono culm 1) mutant is the single-gene recessive mutation, meets Mendelian's mode genetic development.Thereby the quality and yield of regulating paddy rice by the tiller number of cultivation means adjusting and controlling rice has obtained enough attention on producing
(7 8 9)If can realize that the regulation and control to the rice tillering number will be more economical valid approach by biotechnology; Up to now, the rarely seen one piece of report of research of the gene of relevant controlling plant side shoot generation
(13)The present invention passes through the gene M OC1 of the controlled rice tillering proterties of map based cloning technology, and has identified the function of this gene by transgenic function complementation experiment.
Summary of the invention
At above-mentioned research background, the purpose of this invention is to provide a kind of new gene of from rice mutant moc1, cloning, MOC1, the dna sequence dna shown in Fig. 7 and Seq ID No.1 also comprises the gene order that has 70% homology with the dna sequence dna shown in the Seq ID No.1 at least.Seq ID No.2 and protein shown in Figure 8 among the present invention belong to the GRAS transcription factor family, wherein carry out one or several and replace, and insert or lack the functional analogue that is obtained.In addition, be also included within the mutant allele or the derivative that add, replace, insert or delete one or more Nucleotide among the SeqID No.1 and generate, the sequence with identical function also can reach purpose of the present invention.MOC1 and tomato LS albumen homology be 44% with the homology of Arabidopis thaliana Atls be 37%, the function that above-mentioned albumen all has the control side shoot to take place.Therefore, the present invention also comprises the functional analogue that has at least 30% homology with the aminoacid sequence shown in the Seq ID No.2.
Another object of the present invention provides a kind of method of carrying out plant genetic conversion efficiently with MOC1, specifically, the invention provides the gene or the segmental carrier of Gene Partial that have with Seq ID No.1 and sequence shown in Figure 7, wherein, PC8247 as shown in Figure 5, PC8247S, this carrier can express by the polypeptide of above-mentioned nucleic acid sequence encoding or its homology analogue.The present invention also provides a kind of transformant that contains the above expression vector.Transformant comprises intestinal bacteria, Agrobacterium or vegetable cell.
The present invention also provides a kind of plant expression vector transformed plant cells of utilizing to influence that monocotyledons tillers or the method for dicotyledons side shoot generating ability.
Realize that concrete technological step of the present invention is as follows:
One paddy rice singly tiller separation and the genetic analysis of mutant MOC1
By a large amount of screening natural mutants, the present invention obtains the paddy rice mutant MOC1 of singly tillering, by with wild paddy rice reciprocal cross experiment, we obtain a cryptic mutant that meets the genetic development of single-gene control, as shown in Figure 1.
Two, the MOC1 gene of map based cloning control rice tillering
The Primary Location of MOC1 gene
In order to separate the MOC1 gene, the present invention adopts the method for map based cloning, at first sets up a target group that big polymorphism is high, by bright extensive 63 (indica, MOCL/MOC1) X moc1 (japonica, moc1/moc1) F
2In recessive individual the composition.And utilize SSLP, and CAPS and RFLP equimolecular mark carry out Primary Location to the MOC1 site and see Fig. 2. and positioning result shows that the MOC1 Primary Location is long-armed between S1437 and two marks of R1559 at the 6th karyomit(e).
The physical positioning of MOC1 gene
YAC by setting up the MOC1 site areas and BAC clone's contig and separate corresponding YAC and the BAC end carries out Fine Mapping and MOC1 is positioned BAC clones on 4 cA11 and see Fig. 3.
The Fine Mapping of MOC1 gene
Subclone and sequential analysis by to BAC clone 4cA11 develop new CAPS mark MOC1 accurately are positioned (Fig. 4) within the 20kb scope, infer candidate gene by the open reading frame (ORF) of analyzing this section.
The evaluation of MOC1 gene and functional analysis
Pass through transgenic technology, the result shows that the present invention has obtained to make mutant to recover the transgenic paddy rice (see figure 6) of normal function, proved that the present invention has correctly cloned MOC1 gene (Fig. 7), amino acid sequence analysis shows that MOC1 albumen belongs to GRAS transcription factor family (Fig. 8).
Three, the expression of excessive MOC1 gene in paddy rice
By transgenic technology overexpression MOC1 gene, the transgenic paddy rice that acquisition is tillered more, its tiller number are 3-5 times of wild-type paddy rice, prove that the MOC1 gene can promote the increase (Fig. 9) of rice tillering number.
There is the contradiction that cultivated area reduces significantly, population increases substantially at present in China, this output and quality that presses for farm crop in the unit surface arable land increases substantially, wherein reasonably plant plant type and plant population's structure are the important foundations that crop yield and quality improve, and the development of genetic engineering technique makes to use MOC1 gene adjustment plant plant type and form rational plant population structure becomes possibility.
Description of drawings
Be described in further detail understanding the present invention below in conjunction with accompanying drawing, but be not that the present invention is limited.
Fig. 1. singly the tiller phenotype of mutant moc1 of paddy rice.
The Primary Location figure of Fig. 2 .MOC1 on paddy rice the 6th karyomit(e)
The physical positioning of Fig. 3 .MOC1 gene
The Fine Mapping of Fig. 4 .MOC1 gene
Fig. 5 .pC8247S and pC8247 carrier collection of illustrative plates
Fig. 6. function complementation experiment T1 is for the phenotype of transgenic paddy rice. a left side: wild-type H89025; In: the pC8247S plasmid transforms T1 generation; Right: the pC8247 plasmid transforms T1 generation.
The dna sequence dna of Fig. 7 .MOC1 gene
The aminoacid sequence of Fig. 8 .MOC1 genes encoding
Fig. 9. the tiller number of overexpression MOC1 transgenic paddy rice
Embodiment
1, paddy rice (Oryza sativa ssp.Japonica) mutant monoculm 1 (moc1), original open country
Giving birth to the section bar material is H89025
2, the cultivation condition of rice material
Rice paddy seed was soaked in water 3 days, sowed on the seedbed then.The rice seedling of 4 leaf phases is transplanted in the paddy field, divides the individual plant rice transplanting, and density is 18 * 20cm
2For of the influence of research environment factor to tiller development, H89025 and MOC1 be respectively 5 different sowing time, plants under 3 kinds of different nitrogenous fertilizer levels and the 4 kinds of different planting densities, and actual conditions is as follows: (1) sowing time, from April to the August, respectively at every month sowing in first day; (2) the nitrogenous fertilizer level is respectively 150,300,450kg urea/hectare; (3) planting density is respectively 16 * 18, and 18 * 18,18 * 20 and 20 * 20cm
2
3, analysis and target group
Moc1 mutant that isozygotys and original wild-type kind H89025 carry out quadrature and reciprocal cross, F
1For selfing, at F
2The peak period of tillering in generation, the number of statistics wild-type and single tiller phenotype individuality, and calculate segregation ratio with statistical method.F
2Target group is obtained by mutant moc1 (japonica rice) and bright extensive 63 hybridization of rice variety, identifies the F of 2010 single tiller phenotypes altogether
2Individuality has been got tender leaf about 2 grams in the peak period of tillering every strain, is used for extracting total DNA.
4, by SSLP, RFLP and CAPS mark location MOC1 gene
Adopt improved CTAB method
(10)From rice leaf, 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 to and is extracted total DNA in the 1.5ml centrifuge tube, and the DNA resolution of precipitate of acquisition is in 100 μ l ultrapure waters.Each SSLP and CAPS reaction is with 1 μ l DNA sample, and each rflp analysis is with 30 μ l DNA samples.
In the Primary Location stage of MOC1 gene, to carrying out SSLP and rflp analysis by the individual microcommunity of forming of 280 F2.According to forefathers' report, 90 pairs of SSLP primers have been synthesized
(11), and carry out pcr amplification according to the condition of report, separate and EB dyeing the polymorphism of detection PCR product then through 4% agarose gel electrophoresis.When carrying out RFLP (with the Southern of this paper rest part) analysis, use different restriction enzyme complete digestion genomic dnas respectively, separate, transfer on the Hybond N+ nylon membrane (Amersham) through 0.8% agarose gel electrophoresis.With Prime-a-Gene Labeling System (Cat.No.U1100, Promega) and α-32P-dCTP label probe.Nylon membrane after the hybridization detects polymorphism with phosphorus screen molecular imaging instrument (PhosphorImager, Molecular Dynamics).Used RFLP probe, a part come from Nipponbare-Kasalath High Density Molecular linkage map, YAC that another part is separated to when then being physical positioning MOC1 gene and BAC clone's terminal dna fragmentation.
When Fine Mapping MOC1 gene, analyze carrying out CAPS by the individual large group of forming of 2010 F2
(12)According to the sequence of BAC clone 4cA11, we have designed 4 pairs of PCR primers (primer 1:5 ' GACCACTTGATCTCTCATCGAC3 ', 5 ' GAGATCGAACAAGATGGGGAC3 '; Primer 2: 5 ' GCCACTTGATCTCCTAAGTG, 3 ', 5 ' GATGAGACGTCTGATCACAG 3 '; Primer 3:5 ' GTTTGACACTCCCACTGATGG 3 ', 5 ' GGATCATATCCACCATGCATG 3 '; Primer 4,5 ' GTAACGGGAGGTAGCTCTTGAG, 3 ', 5 ' AAAGAATCAAGCAGCAGGTGG 3 ') be that template is carried out pcr amplification with two parent MOC1 and bright extensive 63 genomic dna respectively.Two parents' PCR product with the different restriction enzyme digestion of kind more than 100, separates and bromination second pyridine (EB) dyeing through 2% agarose gel electrophoresis respectively, detects the polymorphism that enzyme is cut product.In case found new CAPS mark, just 2010 F2 individualities are carried out linkage analysis, filter out the individuality that exchange has taken place between this CAPS mark and MOC1 gene.5, the physical positioning of MOC1 gene
In the YAC physical map of the rice varieties Nipponbare that has delivered (http://rgp.dna.affrc.go.jp/Publicdata.html), there are 11 yac clones to be positioned at the MOC1 zone.It is terminal as Y2242R that we adopt the method for iPCR (inverse Polymerase ChainReactions) to separate some YAC, Y2242L, and Y4149R and Y4149L, Y2242R wherein successfully is used as the RFLP probe.Then, use the BAC library (by Clemson University Genomics Institute, CUGI provides) of RFLP probe (R1559 and Y2242R) the Screening of Rice kind Nipponbare of the both sides of MOC1 respectively.The finger printing that the BAC that provides according to CUGI clones is arranged in a BAC contig with these positive colonies.The BAC end sequence (http://www.genome.clemson.edu/projects/rice/rice_bac_end/) that produces according to CUGI STC order-checking plan designs primer, adopt PCR method to separate some BAC ends, wherein two BAC ends are that 18dD02R and 45cD09F successfully are used as the RFLP probe.The BAC clone 4cA11 that comprises the MOC1 gene is broken into the fragment of 1.5-3.0kb with ultrasonic wave, the end-filling rear clone in plasmid vector pUC19, thereby set up the random library of the 4cA11 that is used for the BAC order-checking, and order-checking.6, the prediction of gene and comparative analysis
The genome sequence in candidate zone carries out BLASTX then and analyzes with the possible coding region (ORF) of GENSCAN software (http:/genes.mit.edu/GENSCAN.html) prediction in the Protein Data Bank of ORFs in GenBank with prediction.Carry out protein sequence comparison and evolutionary tree analysis with the Clustal method in DNAStar software (Lasergene) the MegAlign program.According to the sequences Design primer of BAC clone 4cA11, adopt PCR method from the genomic dna of mutant moc1 and wild-type H89025, to amplify the MOC1 gene fragment respectively.To check order respectively from the PCR product of three independent reactions, all contain the retrotransponsons of the 1.9kb in the MOC1 mutant.
Plant Transformation P4123 and P4124 are used for measuring two plasmid clones that BAC clones the random library of 4cA11 sequence, by total SacI site they are connected into a 3.2kb fragment, comprising MOC1 complete ORF district, 1.5kb upstream sequence and 0.3kb downstream sequence.Cut P4123 with the SacI enzyme simultaneously, produce a 2.4kb fragment, only contain the ORF of MOC1 part, fallen 188 amino acid at the C-terminal deletion.3.2kb be cloned into respectively among the binary vector pCAMBIA1300 with the 2.4kb fragment, obtained the plasmid pC8247 and the pC8247S (Fig. 5) that are used to transform.It is rice transformation among the LBA4404 that these two plasmids change Agrobacterium (Agrobacteriumtumefaciens) strain over to by the method that shocks by electricity.Mature seed shelling sterilization with the moc1 mutant is inoculated in the substratum of callus induction.After cultivating for 3 weeks, grow callus from scultellum, it is vigorous to select growth, and color is pale yellow, and more open embryo callus subculture is as the acceptor that transforms.Infect rice callus with the LBA4404 bacterial strain that contains the double base plasmid vector, cultivate after 3 days for 25 ℃, containing screening kanamycin-resistant callus tissue and transfer-gen plant on the selection substratum of 50mg/L Totomycin at the dark place.With hygromycin resistance plant hardening in the cool, be transplanted to paddy field (Fig. 6) after several days.
Moc1?DNA?sequence.ST25
SEQUENCE LISTING＜110〉Chinese Academy of Sciences's heredity and Developmental Biology research institute＜120〉rice tillering controlling gene moc1 and application＜130 thereof〉patent＜160〉2＜170〉PatentIn version 3.1＜210〉1＜211〉1326＜212〉DNA＜213〉Oryza sativa＜220〉＜221〉CDS＜222〉(1) .. (1326)＜223〉＜400〉1atg ctc cgg tca ctc cac tcc tcg tcg tct tct gac acg gat aac aac 48Met Leu Arg Ser Leu His Ser Ser Ser Ser Ser Asp Thr Asp Asn Asn1,5 10 15age ggt ggc tgc aag aac aat ggc ggc ggc ggt ggc gag gcc gcc gcc 96Ser Gly Gly Cys Lys Asn Asn Gly Gly Gly Gly Gly Glu Ala Ala Ala
20 25 30gcc?gtt?gag?ggt?ggc?ggt?gat?cag?cgt?gcc?gtt?gcg?gcg?gcg?gcg?ccg 144Ala?Val?Glu?Gly?Gly?Gly?Asp?Gln?Arg?Ala?Val?Ala?Ala?Ala?Ala?Pro
35 40 45tcg?acg?cgg?gac?ttg?ctg?ctg?gcg?tgc?gcg?gac?ctg?ctg?cag?agg?ggg 192Ser?Thr?Arg?Asp?Leu?Leu?Leu?Ala?Cys?Ala?Asp?Leu?Leu?Gln?Arg?Gly
50 55 60gac?ctg?ccg?gcg?gcg?agg?cga?gcg?gcg?gag?atc?gtc?ttg?gcg?gcg?gcg 240Asp?Leu?Pro?Ala?Ala?Arg?Arg?Ala?Ala?Glu?Ile?Val?Leu?Ala?Ala?Ala65 70 75 80gcg?tcg?ccg?cgg?ggc?gac?gcg?gcg?gac?cgc?ctg?gcg?tac?cacttc?gcc 288Ala?Ser?Pro?Arg?Gly?Asp?Ala?Ala?Asp?Arg?Leu?Ala?Tyr?His?Phe?Ala
85 90 95cgc?gcg?ctg?gcg?ctc?cgg?gtg?gac?gcc?aag?gcc?ggc?cat?ggc?cac?gtc 336Arg?Ala?Leu?Ala?Leu?Arg?Val?Asp?Ala?Lys?Ala?Gly?His?Gly?His?Val
100 105 110gtc?gtc?ggc?ggc?ggc?gcg?gcg?cgg?ccg?gcg?tcg?tcc?ggg?gcg?tac?ctg 384Val?Val?Gly?Gly?Gly?Ala?Ala?Arg?Pro?Ala?Ser?Ser?Gly?Ala?Tyr?Leu
115 120 125gcg?ttc?aac?cag?atc?gcg?ccg?ttc?ctg?cgt?ttc?gcg?cac?ctc?acg?gcg 432Ala?Phe?Asn?Gln?Ile?Ala?Pro?Phe?Leu?Arg?Phe?Ala?His?Leu?Thr?Ala
130 135 140aac?cag?gcc?atc?ctc?gag?gcc?gtc?gac?ggc?gcg?cgc?cgc?gtc?cac?atc 480Asn?Gln?Ala?Ile?Leu?Glu?Ala?Val?Asp?Gly?Ala?Arg?Arg?Val?His?Ile145 150 155 160ctc?gac?ctc?gac?gcc?gtc?cac?ggc?gtg?cag?tgg?ccg?ccg?ctg?ctg?cag 528Leu?Asp?Leu?Asp?Ala?Val?His?Gly?Val?Gln?Trp?Pro?Pro?Leu?Leu?Gln
165 170 175gcc?atc?gcc?gag?cgc?gcg?gac?ccg?gcg?ctc?ggc?ccg?ccc?gag?gtc?cgc 576Ala?Ile?Ala?Glu?Arg?Ala?Asp?Pro?Ala?Leu?Gly?Pro?Pro?Glu?Val?Arg
180 185 190gtc?acc?ggc?gcc?ggc?gcc?gac?cgc?gac?acc?ctc?ctc?cgc?acc?ggc?aac 624Val?Thr?Gly?Ala?Gly?Ala?Asp?Arg?Asp?Thr?Leu?Leu?Arg?Thr?Gly?Asn
195 200 205
Moc1?DNA?sequence.?ST25cgc?ctc?cgc?gcc?ttc?gcc?cgc?tcc?atc?cac?ctc?ccc?ttc?cac?ttc?acc 672Arg?Leu?Arg?Ala?Phe?Ala?Arg?Ser?Ile?His?Leu?Pro?Phe?His?Phe?Thr
210 215 220ccg?ctc?ctc?ctc?tcc?tgc?gcc?acc?acg?gcg?ccc?cac?cac?gtc?gcc?ggc 720Pro?Leu?Leu?Leu?Ser?Cys?Ala?Thr?Thr?Ala?Pro?His?His?Val?Ala?Gly225 230 235 240acg?agc?acc?ggc?gcc?gcc?gcc?gcc?gcc?tcg?acc?gcc?gcc?gcc?gcc?act 768Thr?Ser?Thr?Gly?Ala?Ala?Ala?Ala?Ala?Ser?Thr?Ala?Ala?Ala?Ala?Thr
245 250 255ggc?ctc?gag?ttt?cac?ccg?gac?gag?acg?ctc?gcc?gtg?aac?tgc?gtc?atg 816Gly?Leu?Glu?Phe?His?Pro?Asp?Glu?Thr?Leu?Ala?Val?Asn?Cys?Val?Met
260 265 270ttc?ttg?cac?aac?ctg?gcc?ggc?cac?gac?gag?ctc?gcc?gcg?ttc?ttg?aag 864Phe?Leu?His?Asn?Leu?Ala?Gly?His?Asp?Glu?Leu?Ala?Ala?Phe?Leu?Lys
275 280 285tgg?gtg?aag?gcc?atg?tcg?ccc?gcc?gtg?gtg?acc?atc?gcc?gag?agg?gaa 912Trp?Val?Lys?Ala?Met?Ser?Pro?Ala?Val?Val?Thr?Ile?Ala?Glu?Arg?Glu
290 295 300gca?ggc?ggc?ggc?ggc?ggc?ggc?ggc?gac?cac?atc?gac?gac?ctg?ccg?cgg 960Ala?Gly?Gly?Gly?Gly?Gly?Gly?Gly?Asp?His?Ile?Asp?Asp?Leu?Pro?Arg305 310 315 320cgg?gtc?ggg?gtg?gcc?atg?gat?cac?tac?tcg?gcg?gtg?ttc?gag?gcg?ctg 1008Arg?Val?Gly?Val?Ala?Met?Asp?His?Tyr?Ser?Ala?Val?Phe?Glu?Ala?Leu
325 330 335gag?gcg?acg?gtg?ccg?ccg?ggg?agc?cgg?gag?cgc?ctc?gcc?gtg?gag?cag 1056Glu?Ala?Thr?Val?Pro?Pro?Gly?Ser?Arg?Glu?Arg?Leu?Ala?Val?Glu?Gln
340 345 350gag?gtg?ctg?ggc?cgg?gag?atc?gag?gcc?gcg?gtg?ggg?ccc?tcc?ggc?ggc 1104Glu?Val?Leu?Gly?Arg?Glu?Ile?Glu?Ala?Ala?Val?Gly?Pro?Ser?Gly?Gly
355 360 365cgg?tgg?tgg?cgc?ggc?atc?gag?cgg?tgg?ggc?ggc?gcc?gcc?cgc?gcc?gcg 1152Arg?Trp?Trp?Arg?Gly?Ile?Glu?Arg?Trp?Gly?Gly?Ala?Ala?Arg?Ala?Ala
370 375 380ggg?ttc?gcg?gcg?cgg?ccg?ctc?agc?gcg?ttc?gcc?gtg?tcg?cag?gcg?cgg 1200Gly?Phe?Ala?Ala?Arg?Pro?Leu?Ser?Ala?Phe?Ala?Val?Ser?Gln?Ala?Arg385 390 395 400ctg?ctg?ctc?cgg?ctg?cac?tac?ccg?tcg?gag?ggc?tac?ctc?gtg?cag?gag 1248Leu?Leu?Leu?Arg?Leu?His?Tyr?Pro?Ser?Glu?Gly?Tyr?Leu?Val?Gln?Glu
405 410 415gcg?cgc?ggc?gcc?tgc?ttc?ctc?ggg?tgg?cag?acg?cgc?ccg?ctg?ctc?tcc 1296Ala?Arg?Gly?Ala?Cys?Phe?Leu?Gly?Trp?Gln?Thr?Arg?Pro?Leu?Leu?Ser
420 425 430gtc?tca?gcg?tgg?cag?ccg?tcg?tcg?tcg?tag 1326Val?Ser?Ala?Trp?Gln?Pro?Ser?Ser?Ser
435 440<210>2<211>441<212>PRT<213>Oryza?sativa<400>2Met?Leu?Arg?Ser?Leu?His?Ser?Ser?Ser?Ser?Ser?Asp?Thr?Asp?Asn?Asn1 5 10 15Ser?Gly?Gly?Cys?Lys?Asn?Asn?Gly?Gly?Gly?Gly?Gly?Glu?Ala?Ala?Ala
20 25 30
Moc1?DNA?sequence.ST25Ala?Val?Glu?Gly?Gly?Gly?Asp?Gln?Arg?Ala?Val?Ala?Ala?Ala?Ala?Pro
35 40 45Ser?Thr?Arg?Asp?Leu?Leu?Leu?Ala?Cys?Ala?Asp?Leu?Leu?Gln?Arg?Gly
50 55 60Asp?Leu?Pro?Ala?Ala?Arg?Arg?Ala?Ala?Glu?Ile?Val?Leu?Ala?Ala?Ala65 70 75 80Ala?Ser?Pro?Arg?Gly?Asp?Ala?Ala?Asp?Arg?Leu?Ala?Tyr?His?Phe?Ala
85 90 95Arg?Ala?Leu?Ala?Leu?Arg?Val?Asp?Ala?Lys?Ala?Gly?His?Gly?His?Val
100 105 110Val?Val?Gly?Gly?Gly?Ala?Ala?Arg?Pro?Ala?Ser?Ser?Gly?Ala?Tyr?Leu
115 120 125Ala?Phe?Asn?Gln?Ile?Ala?Pro?Phe?Leu?Arg?Phe?Ala?His?Leu?Thr?Ala
130 135 140Asn?Gln?Ala?Ile?Leu?Glu?Ala?Val?Asp?Gly?Ala?Arg?Arg?Val?His?Ile145 150 155 160Leu?Asp?Leu?Asp?Ala?Val?His?Gly?Val?Gln?Trp?Pro?Pro?Leu?Leu?Gln
165 170 175Ala?Ile?Ala?Glu?Arg?Ala?Asp?Pro?Ala?Leu?Gly?Pro?Pro?Glu?Val?Arg
180 185 190Val?Thr?Gly?Ala?Gly?Ala?Asp?Arg?Asp?Thr?Leu?Leu?Arg?Thr?Gly?Asn
195 200 205Arg?Leu?Arg?Ala?Phe?Ala?Arg?Ser?Ile?His?Leu?Pro?Phe?His?Phe?Thr
210 215 220Pro?Leu?Leu?Leu?Ser?Cys?Ala?Thr?Thr?Ala?Pro?His?His?Val?Ala?Gly225 230 235 240Thr?Ser?Thr?Gly?Ala?Ala?Ala?Ala?Ala?Ser?Thr?Ala?Ala?Ala?Ala?Thr
245 250 255Gly?Leu?Glu?Phe?His?Pro?Asp?Glu?Thr?Leu?Ala?Val?Asn?Cys?Val?Met
260 265 270Phe?Leu?His?Asn?Leu?Ala?Gly?His?Asp?Glu?Leu?Ala?Ala?Phe?Leu?Lys
275 280 285Trp?Val?Lys?Ala?Met?Ser?Pro?Ala?Val?Val?Thr?Ile?Ala?Glu?Arg?Glu
290 295 300Ala?Gly?Gly?Gly?Gly?Gly?Gly?Gly?Asp?His?Ile?Asp?Asp?Leu?Pro?Arg305 310 315 320Arg?Val?Gly?Val?Ala?Met?Asp?His?Tyr?Ser?Ala?Val?Phe?Glu?Ala?Leu
325 330 335
Moc1?DNA?sequence.?ST25Glu?Ala?Thr?Val?Pro?Pro?Gly?Ser?Arg?Glu?Arg?Leu?Ala?Val?Glu?Gln
340 345 350Glu?Val?Leu?Gly?Arg?Glu?Ile?Glu?Ala?Ala?Val?Gly?Pro?Ser?Gly?Gly
355 360 365Arg?Trp?Trp?Arg?Gly?Ile?Glu?Arg?Trp?Gly?Gly?Ala?Ala?Arg?Ala?Ala
370 375 380Gly?Phe?Ala?Ala?Arg?Pro?Leu?Ser?Ala?Phe?Ala?Val?Ser?Gln?Ala?Arg385 390 395 400Leu?Leu?Leu?Arg?Leu?His?Tyr?Pro?Ser?Glu?Gly?Tyr?Leu?Val?Gln?Glu
405 410 415Ala?Arg?Gly?Ala?Cys?Phe?Leu?Gly?Trp?Gln?Thr?Arg?Pro?Leu?Leu?Ser
420 425 430Val?Ser?Ala?Trp?Gln?Pro?Ser?Ser?Ser
435 440
Claims (15)
1. rice tillering or branch controlling gene MOC1 encoded protein matter, or the function albuminoid in other species, the aminoacid sequence shown in its aminoacid sequence and the Seq ID No.2 has at least 30% homology.
2. protein according to claim 1 has the aminoacid sequence shown in the Seq ID No.2.
3. protein according to claim 1, wherein said aminoacid sequence also are included in and add, replace, insert or delete the derivative that one or more amino acid generate in the aminoacid sequence shown in the Seq ID No.2.
4. the gene of the claim 1 of encoding, 2 or 3 any described protein or its functional analogue.
5. gene according to claim 4, it has the nucleotide sequence shown in the Seq.ID.No.1.
6. gene according to claim 4, wherein said nucleotide sequence also are included in mutant, allelotrope or the derivative that add, replace, insert or delete one or more Nucleotide in the nucleotide sequence shown in the Seq.ID.No.1 and generate.
7. plasmid that contains claim 4,5 or 6 described genes.
8. plasmid that contains the partial sequence of claim 4,5 or 6 described genes.
9. one kind is pC8247 or pC8247S according to the described plasmid of claim 8.
10. plant expression vector that contains claim 7,8 or 9 described plasmids.
11. transformant that contains the described gene order of claim 10.
12. according to the described transformant of claim 11, this cell is intestinal bacteria.
13. transformant according to claim 11, this cell are Agrobacterium.
14. transformant according to claim 11, this cell are plant.
15. one kind cultivates plants and tillers or branched method, comprises with claim 10 11,12,13 described expression vectors or host cell transformed plant cells; With the plant transformed cell culture is become plant.
Priority Applications (3)
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| CNB021291969A CN1185256C (en) | 2002-08-20 | 2002-08-20 | Rice tiller control gene MOC1 and its application |
| PCT/CN2003/000429 WO2004018508A1 (en) | 2002-08-20 | 2003-06-03 | A protein controlling rice tiller, a gene encoding the protein and a method of manipulating plant tiller or branching using the gene |
| AU2003245803A AU2003245803A1 (en) | 2002-08-20 | 2003-06-03 | A protein controlling rice tiller, a gene encoding the protein and a method of manipulating plant tiller or branching using the gene |
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| CN100429310C (en) * | 2006-03-15 | 2008-10-29 | 华中农业大学 | Genes controlling tillering in rice and their uses |
| CN101182524B (en) * | 2006-04-28 | 2010-06-16 | 中国农业大学 | A gene regulating rice tiller angle and its encoded protein and application |
| CN1970767B (en) * | 2006-11-29 | 2010-07-14 | 中国科学院遗传与发育生物学研究所 | Gene related to plant type from rice and its encoded protein and application |
| CN101392024B (en) * | 2005-03-31 | 2011-10-05 | 中国科学院遗传与发育生物学研究所 | Rice tiller-related protein and its coding gene and application |
| CN101585869B (en) * | 2009-05-11 | 2012-05-23 | 中国科学院遗传与发育生物学研究所 | A kind of rice tiller-related protein and its coding gene and application |
| CN102533760A (en) * | 2011-11-29 | 2012-07-04 | 中国科学院华南植物园 | Small-molecule ribonucleic acid (RNA) Osa-miR393 for improving rice tillering and application |
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2002
- 2002-08-20 CN CNB021291969A patent/CN1185256C/en not_active Expired - Fee Related
-
2003
- 2003-06-03 AU AU2003245803A patent/AU2003245803A1/en not_active Abandoned
- 2003-06-03 WO PCT/CN2003/000429 patent/WO2004018508A1/en not_active Application Discontinuation
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| CN105462992B (en) * | 2016-01-06 | 2019-03-05 | 河南师范大学 | Rice Os RhoGAP2 gene is improving the application in rice effective tillering |
| CN108085319A (en) * | 2017-12-25 | 2018-05-29 | 中国科学院遗传与发育生物学研究所 | Plant tillering angle GAP-associated protein GAP and its encoding gene and application |
| CN110863006A (en) * | 2019-11-15 | 2020-03-06 | 厦门大学 | A method for improving tillering and regeneration of rice |
| CN110863006B (en) * | 2019-11-15 | 2021-04-30 | 厦门大学 | A method for improving tillering and regeneration of rice |
Also Published As
| Publication number | Publication date |
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| AU2003245803A1 (en) | 2004-03-11 |
| WO2004018508A1 (en) | 2004-03-04 |
| CN1185256C (en) | 2005-01-19 |
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