CN101585869B - Rice tillering associated protein, encoding gene and use thereof - Google Patents
Rice tillering associated protein, encoding gene and use thereof Download PDFInfo
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- CN101585869B CN101585869B CN200910083935XA CN200910083935A CN101585869B CN 101585869 B CN101585869 B CN 101585869B CN 200910083935X A CN200910083935X A CN 200910083935XA CN 200910083935 A CN200910083935 A CN 200910083935A CN 101585869 B CN101585869 B CN 101585869B
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Abstract
The present invention discloses a rice tillering associated protein, the encoding gene and the use thereof. The protein is selected from the following proteins (a) and (b): (a) a protein which is composed of the amino acid sequence represented by the sequence 4 in the sequence table; (b) a protein which is associated with the plant tillering and is derivate from the protein (a) through substituting and/or deleting and/or adding one or a plurality of amino-acid residues to the amino acid sequence represented by the sequence 4 in the sequence table. The transgene complementation test proves thatthe protein D27 coded by the gene participates in the synthesizing of the strigolactone and regulates the plant tillering or the branch stem developing. Therefore, the genetic engineering technology can be used for regulating the tillering trait of the rice, regulating the nutrition using efficiency of the plant and effectively controlling the destructive parasitic weed. The D27 gene has important application value in the aspect of researching the beneficial fungus association and the destructive parasitic weed in the agriculture and natural ecosystem.
Description
Technical field
The present invention relates to a kind of rice tillering associated protein and encoding sox thereof and application.
Background technology
The branch of growing on the paddy rice basal internode is commonly referred to as tillers, and is the branch of paddy rice particular type.Tillering in Rice Production is an important economical character.On the one hand, individual plant effective tillering number has determined the spike number of unit surface, and spike number has determined the output of unit surface; On the other hand, the individual plant tillering number is an important factor of forming plant type structure, and the ideal plant type structure helps ventilation and penetrating light, improves photosynthetic efficiency, absorption and distribution mineral nutrition reasonably, thereby promotes effective tillering and suppress ineffective tillering.In production practice, on the one hand,, take suitable cultivation step to promote to tiller into fringe according to the growth-development law of tillering; On the other hand, has the kind of desirable tillering number according to genetic theoretical the cultivation.Therefore, the research of rice tillering mechanism has important directive significance for agriculture prodn.
Tillering number that it is generally acknowledged paddy rice is the quantitative character that receives a plurality of Gene Handling, is easy to affected by environment.Reported nearly 23 quantitative trait locus (QTLs) that influences the rice tillering number, be distributed in except that on remaining 10 karyomit(e) outside the 9th, the No. 10.In addition, the proterties that the research in past also will be relevant with the rice tillering growth such as the dormancy and the leaf age of rice tillering bud have carried out qtl analysis, and on 12 karyomit(e)s, have all done the location to these related locus.Although the tillering number of paddy rice receives controlled by multiple genes, the unit point of some major genes sudden change just can causing tillering number generation noticeable change.Up to the present, separated and studied the rice mutant that some tillering numbers change.The Japan scientist is with gamma-rays radioinduction rice paddy seed, separates to obtain 5 different two mutants of tillering that lack, and distinguishes called after rcn1 (reduced culm number), rcn2, rcn3, rcn4 and rcn5.These rcn two mutants obviously receive the influence of envrionment conditions, and under high temperature and high nitrogen nutrition condition, its tillering number almost returns to normal level.Infer that in view of the above rcn two mutants tiller bud is differentiated to form process maybe be unaffected, and in growth and development process subsequently, have defective, causes tiller bud not extend, thereby cause that tillering number reduces.Genetic analysis shows that the rcn two mutants is by single recessive gene control, and rcn1, and rcn2, rcn5 are positioned at respectively on the 6th, 4, No. 6 karyomit(e)s of paddy rice.Belong to together in the Gramineae barley in paddy rice, also found some tiller less two mutants such as uniculm and rnt.The another kind of two mutants of tillering of paddy rice, showing as the individual plant tillering number increases.At least 6 nonallelic Dwarf Mutant are that d3, d10, d14, d17, d27 and d53 show very strong tillering ability, and these two mutants plant is short and small grows thickly.Through genetic linkage analysis, the gene locus of controlling these proterties is positioned near the relevant chromosomal marker site mostly.Wherein, D3, D10 and D17 gene are cloned in succession.The result of study of said gene shows that the rice tillering adjusting and controlling growth relates to very complicated phytohormone Regulation and signal transduction pathway.
Recently; The hormone or the only angle gold lactone of hormone synthetic precursor substance (strigolactones) of one type of new regulation and control plant branching growth are identified out; Type of belonging to terpene lactones is secreted out from plant roots on its chemical structure, can prevent the excessive branch of plant.Solely gold lactone in angle at first is studied as the seed germination accelerant of root parasitism weeds; The whole world has more than 3000 kind of phytoparasite approximately; They are distributed widely in the various ecological, and its denominator is must from the host, rob some nutritive substance and moisture etc. to satisfy the needs of self giving birth to.Phytoparasite has or not according to its plant is chlorophyllous, is divided into semiparasitic plant and holoparasite; Different according to its parasitic site simultaneously, can be divided into root parasitism plant and cauline leaf phytoparasite again.Wherein Orobanche and unipods metal are respectively holoparasite and semiparasitic plant, and both are of paramount importance root parasitism weeds in the phytoparasite, and at aspects such as its fertility and parasitic habits many similaritys are arranged.Comprise two main life stages the life history of root parasitism plant: autotrophy stage and parasitic stage.Broomrape and witchweed seed are through behind the preparatory cultivation stage; Under the inducing of the only angle of host's root system excretory gold lactone, sprout; To the host plant root, this moment, parasite just began to be changed to the heterotrophic growth stage (parasitism) by autophyting growth the radicle of sprouting the back seed through the haustorium parasitism, promptly began to be adsorbed on host's root and penetrated host's root; Connect through vascular bundle and to absorb its nutritive substance and moisture etc., until parasite plant unearthed, the maturation of blooming, set seeds of growing up with host's root.Other discovers that this natural sprouting stimulator can be convened the fungal component that helps plant absorbing nutrition, and this discovery is given prominence to and shown the substantial connection between plant and the fungi.Though the synthetic and mechanism of action about only angle gold lactone still awaits further clear and definite at present.But solely this new plant branching controlling elements of angle gold lactone provide a new research direction for the control of useful mycosymbiosis body and destructive parasitic weeds in research agricultural and the natural ecosystems.
Summary of the invention
An object of the present invention is to provide and control relevant albumen and encoding sox thereof a kind of tillering with plant.
Protein name provided by the present invention is D27, derives from paddy rice, be specially as follows (a) or (b) shown in albumen:
(a) protein of forming by the aminoacid sequence shown in the sequence in the sequence table 4;
(b) with the aminoacid sequence of sequence in the sequence table 4 through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and tiller relevant by (a) deutero-protein with plant.
The replacement of said one or several amino-acid residue and/or disappearance and/or interpolation are meant replacement and/or disappearance and/or the interpolation that maintenance and albumen shown in the sequence 4 have 30% above homology.
(b) proteic aminoacid sequence described in specifically can be shown in sequence in the sequence table 5.
For make (a) or (b) in albumen be convenient to purifying, can connect label as shown in table 1 at said proteic N end or C end.
The sequence of table 1. label
| Label | Residue | Sequence |
| Poly-Arg | 5-6 (being generally 5) | RRRRR |
| Poly-His | 2-10 (being generally 6) | HHHHHH |
| FLAG | 8 | DYKDDDDK |
| Strep-tag II | 8 | WSHPQFEK |
| c-myc | 10 | EQKLISEEDL |
Above-mentioned (a) but or the albumen synthetic (b), also can synthesize its encoding sox earlier, carry out biology again and express and to obtain.Proteic encoding sox in above-mentioned (b) can be through the codon with one or several amino-acid residue of the disappearance of the dna sequence dna shown in sequence in the sequence table 1,2 or 3; And/or carry out the missense mutation of one or several base pair, and/or obtain at the encoding sequence that its 5 ' end and/or 3 ' end connects the label shown in the table 1.
Replacement, replacement and/or the interpolation of one or several amino-acid residue in the above-mentioned proteic aminoacid sequence are had plenty of because abiogenous variation causes, have plenty of to be handled by induced mutations to cause.
Encoding sox provided by the present invention specifically can be following 1) or 2) or 3) or 4) or 5) or 6) shown in gene:
1) in the sequence table sequence 1 from 5 ' dna molecular shown in the Nucleotide of terminal 2237-7205 position;
2) dna molecular shown in the sequence 1 in the sequence table;
3) dna molecular shown in the sequence 2 in the sequence table;
4) dna molecular shown in the sequence 3 in the sequence table;
5) under stringent condition with 1) or 2) or 3) or 4) dna molecule hybridize that limits and the dna molecular of the said plant tillering associated protein of encoding;
6) with 1) or 2) or 3) or 4) dna sequence dna that limits has the dna molecular of the 70% above homology and the said plant tillering associated protein of encoding.
Above-mentioned stringent condition can be 0.1 * SSPE (or 0.1 * SSC), in the solution of 0.1%SDS, under 65 ℃, hybridize and wash film.
The recombinant vectors, reorganization bacterium, transgenic cell line or the expression cassette that contain above-mentioned arbitrary said encoding sox also belong to protection scope of the present invention.
Increase above-mentioned arbitrary said encoding sox total length or its any segmental primer to also belonging to protection scope of the present invention.
Said primer is to specifically can be following 1), 2) or 3):
1) sequence of a primer is shown in sequence in the sequence table 6, and the sequence of another primer is shown in sequence in the sequence table 7;
2) sequence of a primer is shown in sequence in the sequence table 8, and the sequence of another primer is shown in sequence in the sequence table 9;
3) sequence of a primer is shown in sequence in the sequence table 10, and the sequence of another primer is shown in sequence in the sequence table 11;
The application of above-mentioned arbitrary said albumen in the adjusting plant tillers also belongs to protection scope of the present invention.
The application of above-mentioned arbitrary said encoding sox in the adjusting plant tillers also belongs to protection scope of the present invention.
The application in the gold lactone content of only angle in regulating plant of above-mentioned arbitrary said albumen also belongs to protection scope of the present invention.
The application in the gold lactone content of only angle in regulating plant of above-mentioned arbitrary said encoding sox also belongs to protection scope of the present invention.
Said plant specifically can be paddy rice.
Last purpose of the present invention provides the tiller method of the plant of increasing of a kind of cultivation.
The tiller method of the plant of increasing of cultivation provided by the present invention is with the above-mentioned arbitrary said encoding sox deactivation that contains in the plant, obtains the plant that tillers and increase.
Wherein, said deactivation specifically can be disturbed through RNA and realize.Said plant specifically can be paddy rice.
The present invention utilizes the method for map based cloning from the short bar of paddy rice is tillered two mutants d27 more, to clone and identified control tiller number goal gene D27.The protein D 27 of this genes encoding of transgene complementation test proof is participated in the synthetic of only angle gold lactone in the paddy rice body, regulate and control tillering or the growth of side shoot of plant.Solely gold lactone in angle is one type of new branch regulatory factor, can promote the germination of root parasitism weed seed, convenes the fungal component that helps plant absorbing nutrition.Therefore, can utilize the proterties of tillering of genetic engineering technique adjusting and controlling rice, regulate the plant nutrition utilising efficiency and effectively control destructive parasitic weeds; Has very important using value aspect the control of D27 gene useful mycosymbiosis body and destructive parasitic weeds in research agricultural and natural ecosystems.
Description of drawings
Fig. 1 is the tiller phenotype of two mutants d27 of paddy rice more.
Fig. 2 is the transgenic paddy rice phenotype of pD27C carrier collection of illustrative plates and function complementation experiment.
Fig. 3 is a RNA interferential transgenic paddy rice phenotype.
Fig. 4 is the mass spectroscopy paddy rice solely content of angle gold lactone of tillering among the two mutants d27 more.
Embodiment
Employed experimental technique is ordinary method like no specified otherwise among the following embodiment.
Used material, reagent etc. like no specified otherwise, all can obtain from commercial sources among the following embodiment.
Rice varieties Shiokari and dwarf27 (being called for short d27) (Ishikawa S; Maekawa M, Arite T, etal.Suppression of tiller bud activity in tillering dwarf mutants of rice.PlantCell Physiol.2005; 46,79-86.).
Carrier pTCK303 (the Wang Z that has the Ubiquitin promotor; Chen C; Xu Y; Et al.A practicalvector for efficient knockdown of gene expression in rice (Oryza sativa L.) .PlantMol.Biol.Rep.2004,22,409-417.).
Paddy rice obtains according to following method cultivation among the following embodiment: the field cultivation of (1) rice material: after rice paddy seed was soaked seed 2 days in water; Moved between 37 ℃ of cultivations vernalization 3 days; The seed that will show money or valuables one carries unintentionally is then broadcast and on the seedbed, is carried out seedling, rice seedling is transplanted into the paddy field during phase to 4 leaves.(2) water planting of rice material is cultivated: with reference to method (the Kamachi K of Kamachi etc.; Yamaya T; Mae T; Et al.A role for glutaminesynthetase in the remobilization of leaf nitrogen during natural senescence inrice leaves.Plant Physiol.1991,96,411-417.).Rice paddy seed shells the back with 70% ethanol disinfection 1 minute, sterilizes 2 hours with 30% commercially available sodium hypochlorite solution again, and after aqua sterilisa rinsing 6 times, program request is on the nylon wire that floats on water planting solution.Water planting solution was changed once in per two days.
The discovery of embodiment 1, albumen and encoding sox thereof
The tiller seed of two mutants dwarf27 (d27), the seed of wild-type Shiokari of the short bar of paddy rice (Oryza sativa L.) cultivated according to above-mentioned field cultivation method respectively more; The form of seedling (A representes wild-type Shiokari, and B representes the short bar two mutants d27 of tillering more) as shown in Figure 1.Get blade then and carry out the extraction of DNA.
The extraction of oryza sativa genomic dna:
Adopt improved CTAB method (Mou Z; He Y; Dai Y; Et al.Deficiency in fatty acidsynthase leads to premature cell death and dramatic alterations in plantmorphology.The Plant Cell.2000,12,405-418.) from rice leaf, extract genomic dna.Get the 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 MQ H
2Among the O.
Carry out map based cloning according to following steps:
1, the Primary Location of D27 gene (through SSR and CAPS mark location D27 gene)
Because the D27 gene on the 11st karyomit(e) is long-armed, is at first chosen the linkage analysis that some SSR primers carry out goal gene by Primary Location on the 11st karyomit(e), find target gene and little satellite RM206 mark close linkage.According to the est sequence design data primer that this mark near zone has been announced, amplification two mutants and wild type gene group are found STS mark C189 and this gene linkage, and target gene is positioned in zone between these two marks, and genetic distance is 3.1cM.
2, the Fine Mapping of D27 gene
In order further to dwindle the delimited area of goal gene, enlarge target group and developed more CAPS mark.At first, match and made up d27-ZF802/ZF802, d27-ZF802/ Lian Tang morning and three cross combinations of the d27-ZF802/ B of association.F from these three combinations
2Segregating population screens more than 6000 mutation type surface individual plant and is used for the gene linkage analysis.Secondly, 93-11 that has announced through comparison and Japan fine between positioning area in indica rice subspecies genome sequence, seek the difference site, develop the CAPS mark.At F
2When carrying out the linkage analysis of mark and purpose proterties in the two mutants colony; At first with the individual plant that exchange takes place between mark RM206 and C189 screening and the goal gene; With new CAPS mark these exchange individual plants are carried out linkage analysis then; Find molecule marker P3 and P6 and goal gene close linkage at last, and 3 and 1 exchange individual plant are arranged respectively between the goal gene.Goal gene is positioned in the 18kb interval that two CAPS marks on the BAC3 define the most at last.
3, the evaluation of candidate gene and comparative analysis
In order to confirm candidate gene, measured two mutants and wild type gene sequence in the locating area, sequence alignment finds that 4 base deletions of AAGG have taken place the genome sequence of the two mutants in locating area.For clear and definite this 4 base deletion causes mutation type surface; 8 wild-type kinds and 6 corresponding sequences of mutant phenotype individual plant are increased and check order; The result shows that all wild-type materials are all normal in this site, and be total to isolating exchange individual plant with mark P3 and P6 4 base deletions has taken place all in this site.This this site mutation of explanation causes two mutants to produce the phenotype of tillering more.Predictive genes software GENESCAN analysis revealed has two ORFs in locating area 18-kb interval, the mutational site occurs in one of them ORFs, therefore this gene is confirmed as candidate gene.
4, the acquisition of gene order
Candidate gene does not have corresponding full length cDNA sequence in the KOME DB; In order to confirm that this genetic expression and base mutation occur on the exon of gene; Article one chain with the cDNA of wild-type kind Shiokari seedling is a template, adopts Clonetech RACE technology to be increased in the candidate gene cDNA 5 ' upper reaches and 3 ' downstream non-translational region sequences.The RACE experimental result shows, the long 217bp of 5 ' upper reaches non-translational region sequences of this gene, and the long 200bp of 3 ' downstream non-translational region sequences, whole cDNA total length is 1,254bp.Comparison full-length cDNA and genome sequence find that this gene has 7 exons and 6 introns.Deletion mutantion occurs on the 4th exon of this gene, finally causes the protein translation premature termination.
The function of embodiment 2, gene and application
One, the preparation of gene
Extract the mRNA of wild-type kind Shiokari seedling; Reverse transcription obtains cDNA; With primer D27F/D27R is carried out pcr amplification,, obtain the cDNA sequence of D27 gene in the wild-type kind the product order-checking; Its nucleotide sequence is shown in sequence in the sequence table 2, and the aminoacid sequence of its encoded protein is shown in sequence in the sequence table 4.
Extract the mRNA of d27 two mutants seedling; Reverse transcription obtains cDNA; With primer D27F/D27R is carried out pcr amplification,, obtain the cDNA sequence of D27 gene among the two mutants d27 the product order-checking; Its nucleotide sequence is shown in sequence in the sequence table 3, and the aminoacid sequence of its encoded protein is shown in sequence in the sequence table 5.
D27F:5 '-ATGGAGACCACCACGCTTGTGC-3 ' (sequence 6),
D27R:5 '-TCAGATGGAGCAATTCACACC-3 ' (sequence 7).
Compare with the cDNA sequence of D27 gene in the wild-type paddy rice, among the two mutants d27 cDNA sequence deletion of D27 gene 4 Nucleotide (its base is AAGG), deletion mutantion occur in sequence 4 from 5 ' terminal 557-560 position.Deletion mutantion occurs on the 4th exon of gene, finally causes the protein translation premature termination.
Two, functional complementation experiment
Binary vector pCAMBIA1300 is available from Takara company; Agrobacterium (Agrobacterium tumefaciens) strain is that EHA105 is available from Takara company.
To the BAC clone OSJNBa0029K08 (country of Chinese Academy of Sciences cara gene) that contains full-length gene.Carry out the restriction enzyme mapping analysis, find that this BAC clone can obtain a fragment that contains full-length gene through restriction enzyme BamHI and KpnI digestion.This fragment comprises the gene start codon ATG upper reaches 2,236bp and terminator codon TGA downstream 2,044bp, totally 9,249bp (its nucleotide sequence is shown in sequence in the sequence table 1).This fragment subclone to the binary vector pCAMBIA1300 with identical restriction enzyme site, is obtained recombinant plasmid pD27C (Fig. 2 A); Changing plasmid pD27C over to Agrobacterium (Agrobacteriumtumefaciens) strain through the method that shocks by electricity is among the EHA105, and screening obtains containing the reorganization agrobacterium strains of recombinant plasmid pD27C.
Transform for the ease of rice material, the d27 two mutants of the Japanese fine background that we obtain through the mode of backcrossing is the mature seed of this two mutants shelling sterilization; Be inoculated in the substratum of callus induction, cultivated for 3 weeks after, grow callus from scultellum; It is vigorous to select growth; Color is pale yellow, and more open embryo callus is as the acceptor that transforms.
Reorganization agrobacterium strains with containing recombinant plasmid pD27C infects the rice callus tissue, cultivates after 3 days for 25 ℃ at the dark place, is containing screening kanamycin-resistant callus tissue and transfer-gen plant on the selection substratum of 50mg/L Totomycin.The hygromycin resistance plant is refined seedling in the cool, after be transplanted in the paddy field, the transfer-gen plant of acquisition is T
0Generation, results T
0The seed of Dai Miao is cultivated according to the method for above-mentioned field cultivation, and through conventional Molecular Detection, obtains T
1For transgenic seedling.Observe the situation of tillering of transfer-gen plant, 6 T of acquisition
0Tiller number and plant height for transgenic line and offspring's transfer-gen plant thereof all return to the wild-type level.Many tillers two mutants commentaries on classics of short stem D27 gene plant is tillered and having complementary functions of plant height confirms the plant height of D27 Gene Handling paddy rice and growth (Fig. 2 B of tillering; 1 expression wild-type Japan is fine; 2 expressions change the d27 two mutants of empty carrier pCAMBIA1300 over to, and 3 expressions change the T of recombinant vectors pD27C over to
1For plant).
Three, RNA interference experiment
1, the acquisition of interference fragment
CDNA with the Shiokari seedling is a template, with primer D27RNAi1F/D27RNAi1R and primer is carried out pcr amplification to D27RNAi2F/D27RNAi2R respectively, and the product of acquisition checks order.Two pairs of primers increase respectively the nucleotide sequence of gene fragment in the product that obtains like sequence in the sequence table 2 from shown in 5 ' terminal the 164th-820 Nucleotide, long 657bp.
Table 2, primer sequence
| The primer title | Primer sequence |
| D27RNAi1F | 5’-AAGAGCTCCCAAGAAGACGGAGACGGC-3’ |
| D27RNAi1R | 5’-AAACTAGTCACCATGATTCTGCTTTGCG-3’ |
| D27RNAi2F | 5’-AAGGATCCCCAAGAAGACGGAGACGGC-3’ |
| D27RNAi2R | 5’-AAGGTACCCACCATGATTCTGCTTTGCG-3’ |
The front and back primer 5 ' end of D27RNAi1 adds SacI and SpeI joint respectively, and the front and back primer 5 ' end of D27RNAi2 adds BamHI and KpnI joint respectively.
This D27 gene 657bp fragment confirms in rice genome, not have other homologous sequence through full genome compare of analysis.
2, the structure of interference carrier D27-RNAi:
The primer product that amplification obtains to D27RNAi1F/D27RNAi1R is carried out enzyme with SacI and SpeI cut, insert SacI and the SpeI site of the carrier pTCK303 that has the Ubiquitin promotor, obtain carrier 1; The primer product that amplification obtains to D27RNAi2F/D27RNAi2R is carried out enzyme with BamHI and KpnI to be cut; Insertion obtains the BamHI and the KpnI site of carrier 1; Obtain recombinant expression vector D27-RNAi (being interference carrier D27-RNAi), form hairpin structure behind the insertion fragment expression.
Changing interference carrier D27-RNAi over to Agrobacterium (Agrobacteriumtumefaciens) strain through the method that shocks by electricity is among the EHA105, and screening obtains containing the reorganization agrobacterium strains of interference carrier D27-RNAi.
The mature seed shelling sterilization that wild-type paddy rice Japan is fine is inoculated in the substratum of callus induction, cultivated for 3 weeks after, grow callus from scultellum, it is vigorous to select growth, color is pale yellow, more open embryo callus is as the acceptor that transforms.
Reorganization agrobacterium strains with containing interference carrier D27-RNAi infects wild-type rice callus tissue.Cultivate after 3 days for 25 ℃ at the dark place, containing screening kanamycin-resistant callus tissue and transfer-gen plant on the selection substratum of 50mg/L Totomycin.The hygromycin resistance plant is refined seedling in the cool, after be transplanted in the paddy field, the transfer-gen plant of acquisition is T
0In generation, observe T
0The situation of tillering of Dai Miao.The result shows, with Japan fine in behind the D27 gene RNAi, phenotype that short life tillers more obviously appears in transfer-gen plant, and (Fig. 3, A represent that wild-type Japan is fine, and B representes to change over to the T of D27-RNAi
0For transfer-gen plant).
Four, the paddy rice solely content of angle gold lactone (strigolactones) of tillering among the two mutants d27 is identified in systems analysis more
Reference literature (Umehara M; Hanada A, Yoshida S, et al.Inhibition of shoot branchingby new terpenoid plant hormones.Nature.2008; 455,195-200.) said method is carried out the extraction and the LC/MS/MS mensuration of only angle gold lactone.
The experiment material of this experiment usefulness is the tiller seed of two mutants d27, the seed of wild-type kind Shiokari of the short bar of paddy rice more.
Mainly with reference to method (the Kamachi K of Kamachi etc.; Yamaya T; Mae T; Et al.A role forglutamine synthetase in the remobilization of leaf nitrogen during naturalsenescence in rice leaves.Plant Physiol.1991,96,411-417.) method is carried out water planting with rice paddy seed.Choose the hydroponics growing rice material in 3 to 4 weeks, thoroughly clean rice root, place freshly prepared water planting solution to cultivate, collect water planting solution after 24 hours with tap water; Three times (promptly the ETHYLE ACETATE with 1/3 volume extracts water planting solution with equal volume of ethyl acetate; Collect ETHYLE ACETATE phase I, water planting solution is extracted once more collection ETHYLE ACETATE phase II with the ETHYLE ACETATE of 1/3 volume; ETHYLE ACETATE with 1/3 volume extracts water planting solution for the third time; Collect ETHYLE ACETATE phase III), the ETHYLE ACETATE of three collections is merged mutually, use the K of the 0.2M of 1/5 volume again
2HPO
4The ETHYLE ACETATE phase is collected in washing, adds anhydrous Na
2SO
4To remove moisture in the ETHYLE ACETATE, through the rotary evaporation in vacuo appearance ETHYLE ACETATE is revolved driedly, sample is dissolved in 50% acetonitrile, and LC-MS/MS measures.
The condition that LC-MS/MS measures is:
Waters performance liquid chromatography-mass spectrometer system (Quattro Premier XE, Waters MStechnologies, Manchester, UK, Acquity UPLCTM, Waters, Milford, MA, USA).
Chromatographic separation condition:
Chromatographic column: Waters BEH-C
18Post, 2.1 * 50mm, particle diameter 1.7 μ m; Column temperature: 25 ℃; Sample temperature: 25 ℃; The eluent gradient elution requirement: moving phase is distinguished linear growth to 40% and 70% 6 with 15min by the aqueous solution of 30% acetonitrile.Flow velocity: 0.4ml/min.
The mass spectrum condition:
Desolventizing airshed (Desolvation Gas FLow): 800L/hr; Capillary voltage (CapiLLary): 3.80kV; Taper hole voltage (Sample Cone): 30V; Desolventizing temperature degree (Desolvation Temperature): 350 ℃; Ion source temperature (Source Temperature): 120 ℃; Collision voltage (collision energy): 15V, epi-5DS MRM detects ionic reaction passage 331.16＞216.10.Under as above experiment condition, the RT of epi-5DS standard substance is 9.14min.
3 repetitions are established in experiment.
The present invention utilizes ETHYLE ACETATE through liquid-liquid extraction mode separation and Extraction strigolactones from rice root secretory product; And measure the content that the 2 '-epi-5-deoxystrigol (epi-5DS) that clearly identifies analyzes strigolactones through LC/MS/MS; The result is illustrated in the short bar of paddy rice and tillers more and detect less than epi-5DS (Fig. 4 in the root exudates of two mutants d27; The detected result of 1 expression epi-5DS standard substance wherein; 2 expression wild-type kind Shiokari, the 3 expression paddy rice two mutants d27 of tillering more).
More The above results shows tillers among the two mutants d27 the short bar of paddy rice solely the content of angle gold lactone reduces, and prove that paddy rice D27 gene participates in synthesizing of only angle gold lactone.
Sequence table
< 110>Inst. of Genetics and Development Biology, CAS
< 120>a kind of rice tillering associated protein and encoding sox thereof and application
<130>CGGNARC92262
<160>11
<210>1
<211>9249
<212>DNA
< 213>paddy rice (Oryza sativa L.)
<400>1
ggatccttac ttatttctct ccaactctta tttactttcc tctgacttta agttacttac 60
atgtgggctt gttggataga ggattagaga actcatattt actttcctct aactttgagt 120
tactgacatg tgggctcgtt ggatagagga ttagagggtc catattttag tgactcgcgt 180
caaaggggga gtatttcaat ccgttgctgc acgcattgtt ttcgtttgca gtacaaacga 240
aatgcagctg ccgacggttg cctctgataa gatgccacta tcagaattga tgatggcaat 300
agcatatttg tgtaatgctc ccagattgtc ccaacttttt cgcaaactat tctattgttt 360
tttcttttcc tattacccga aagacgcgca tacaacactt gcttgaaaag atggaggtta 420
atggtaaatc tcttatttgg ctaaatatct aataaaaact catatttatg cgtaagattg 480
taggcactaa aatttaaacc agagtggaac catacattgc ttttccaaac tttttttttg 540
aagacattga acttttccaa actgttttgt tttcggctaa ttgtgttgag attttcttct 600
tttggattgg aagacacgtc taagagaatt ttagagtcta gcggcgacgg cttacaagaa 660
agaaaaaaaa agctcctcga agcatccaca taatctaatc tgtcggttac tgcagatcgg 720
acggtcaaaa acggcctaac gagacactgt gcactgcaga tgatctgttt ccctcttacc 780
aggtgctata cagaaattac tgtttgatga tagcttcgtt tgcacctatg tccaaatgat 840
agtatttcac attatttaaa acaattgaag tgtttatatt gaaatactcc atccatcggt 900
tcacaattgt agtaagacaa ttgacaagaa tgactctatt aagactaaga acattggtaa 960
tgtgaagtcg ataccatatt gaagaaaaaa aatgccgatg ccatgccatg gcatgagagc 1020
tttttttttt tttttcacag acattttcat agaaaatgtg gtggcaaaat tcaaataacg 1080
cggggagagt gaaactattt gtggaacaaa gtttaattag ggttaacttt tcagagggga 1140
aaaaatgtaa ctcattaata attttgccac tataatttta ctgttcgctc atttcagtcg 1200
ctaaaaatta gtgctagtcc aaattaaatc ccaatagata gatagtgcaa tttgatttat 1260
tataataaaa ctaaactata tggcattata tccagatggt tcagaccaag ttggtactgt 1320
tcaacttaat gaaactagga ataaactatg atactataac ttcgagatag ttcagacaga 1380
cttggagcga attaactaag aatatacttg agacttgaga gtgaacaaag ctttgtctta 1440
ctagtgagat tatagttagt tacgcgatta tcttggacga cactgattaa tacttcattc 1500
caagttgaca caagattgga taaaaaaaac tcactccatg tagagaatgt taattaaaat 1560
atctaaccaa gctgcctgga tcttttcgcc agctgaccaa ttaaaagaag gaagcatcaa 1620
taaaaaccaa agcccattag agatggacag ctccttgtcc atatatatac tccctctgta 1680
ctcgtaaaag gaagtcgttt tgaacagtga cacggtctcc aaaatataac tttgacttct 1740
tgtttttata aaaatattta ttaaaaagtg atatatgtat acttttatga aagtattttt 1800
caagaaaaat ctattcatat aattttcaca ttttcaaact caacaacttg agagttattt 1860
attatttata ttcctaaggt ttgacttaaa cattgttcta aacgatttct tttacgagta 1920
cggagtactg agggggtata ttccaattag ctatcaacat actaattaac tatcaacctt 1980
aataaaaaca taattattag acactttgat tccttattcc caccacaacc aagatgccct 2040
ctccatgcca tttggtctct tctctctccc tccttgcaaa ttgcatgacc ctctctctct 2100
ctctccactt ctctctataa accttcctct ctcccataac ttctttccat tttcaaccta 2160
caaatatact aatctctctc tagctagtct tcacctacaa atctctctct ctctctctct 2220
ctctctctct ctcttcatgg agaccaccac gcttgtgctg cttcttcctc atggcggcgc 2280
cggcggcgta cggccggcgg cagcggcaac ggcgaagcga agctacgtga tgaggaggtg 2340
ttgctcgacg gtgagggcgg tcatggcgag gccgcaagag gcgccggcgt cggcgccggc 2400
caagaagacg gagacggcgg cgatgatgtc gacggtgcag acggagacgg cggcggcgcc 2460
gccggcgacg gtgtaccggg acagctggtt cgacaagctc gccattggtt acctgtccag 2520
gaaccttcaa gaagcttctg gttcgtgcct ataatctact gaatatatct gatgtattgt 2580
gtgcatgtac atgcaatctt tcggatgaat taatgaatga tcaaatcaac catgcatgca 2640
tatatgtgtg tatttatgat gtttttgcca tggctgattt gattttttga gtggaagaaa 2700
taacatatat atggtgaata aaccaatgtt cttcagttat ttttgtgaca attaaattat 2760
atgtgtattc tacgatcatc cttttcttct tatcgtttgg ttattaatac ccttctcgtc 2820
catgtggaac ataaattctt tgattttttt tattacatgt tgctttggcc agcgtgggtg 2880
tacttgcaat tgcatgcttt ctgtcatatt ataattgaat tagctcttga tttttacatg 2940
caaaagaata atttatttat tgagttgctg atgatcacgt aagcctatgc atgtgagatg 3000
tgatgtgttc tccggcctaa ttaagtatcg acgtactata tatatgcacg tacatcaagt 3060
acggccgtat ttcataattt gattattgtt actgtaccag acaaacctag tgcttaatta 3120
ttgacatata cccaacatcg attaaagtca gtgtgacaca attggcagaa gaagataaca 3180
aaaatcatag aagacatata taccacaaca gtaaaaatta cagtgcaaaa agggatacca 3240
gaagcttctt aattaggtga atggtcacat ttagtagtag taatacacag taataacaca 3300
ttatttgacg aataaattaa gcggaagtgc atgatcagca gcatataagc aagcaactac 3360
gagaggtcaa ccgttgatcc gtgaaaatgt gaggttcaca ttctaggaat cttttagtgc 3420
ttgaatctaa tcctttcgtt tgttttcccg tggacttttt actttgatcg aataaactta 3480
tctgaataat gcgaaggtca aacatcacat gtacgtgccc ggttcttctt ctagaaattc 3540
agccgcttac tataatcgat gctggttttt gaattttttt aaagacattt attttggaac 3600
aacttaaaaa aagtgcacaa accagggttt accaaaccgt ttgaagtgag gttactgcca 3660
ctccatggtt tggcaattat cacgaggtgc gtcgtggttc caaaaactaa taaggttatc 3720
gtatgtgata accgcgataa ctgacggttt tgtaaaccct gcacagacaa cataattata 3780
tttacaggac tgtttggact tcattaacac tgtagcatcc actacatccg tcacctcctt 3840
ggctctctta catgtttagc aaagtcaatc atacagacaa accagaaaag ttcccaacag 3900
acacagaatg ctacttttct agtatgggta attaagcata taagagaaac ttgaaagcat 3960
acaggaaacc gatgttcact agtcctgagt ttttttttgg tgttgttcag gtgcaaatta 4020
actgcaaagg ttgattccct gaaaggaact tgtagcactt gctgactgac acgaacatgc 4080
atgcaatgta atgcagggct aaagaatgaa aaggatggct acgagagcct gatagatgcc 4140
gccctagcca tctcaagaat cttcagtctg gataaacaaa gcgagattgt gacccaagct 4200
cttgaaagag cacttccaag ctacatcctc acaatggtaa gtaccataat ccatgacaat 4260
tggcaatcat gtatgaatta ttgaaattta aaaaacctaa aacttttttt tatttgaaca 4320
ggaagtaaaa ttcagcattt attctctctt ttttttttga ggaaaataca gaacttctgt 4380
aaagggagga aactttaaaa gttctcttct tttatttctt acctcaactt tgattgagaa 4440
ttctgtcggc agatcaaggt gatgatgcca ccttcaagat tttccaggga gtactttgct 4500
gcattcacca cgatattttt tccttggttg gttgggccgt gtgaggtata tattacatac 4560
acagttcctc cttttgttac ttcagttcag aaaggaatag ctgcctgata cctgattagt 4620
gcgtcttcac aagaatgaat tcatgattgt gcctctgctg aaggtagccc tgcagaatga 4680
attgatacga agccacacgt taatttgaga aatattgcta cagaagatct taaaatgctg 4740
ttgaatgtag gttgcgacat ataatatctc tttgttttca ggacattaca ttgtttaaag 4800
tagttgtacg aagcatttgt ggtgcaaata tcatataaag tatggaataa atgcccttgt 4860
atggataact tgtcttctga gtgatcattc tgtcattgaa caaaggttat ggaatctgaa 4920
gttgaaggaa ggaaagagaa aaacgtggta tatatcccca aatgcaggta attcaatcat 4980
catcaagaaa ttgtttcaca agtttgacta ggaggaacat ttgtaaagaa aatttcttca 5040
actgctgggt atcacgtaac atatcggtca aacggtttcc tactcccagc tcttcatagg 5100
cagactacaa aaattttcag gtctataagt agtaacctga tgatttgcaa ttaacatatg 5160
actctgcagc catcaattag ttctgaatga attgttgcag tatgtcaact cttactcaag 5220
ttaatccact catgaaaaac aaattgaaga actatggctg ggacacaagt tgtattgtca 5280
acgtcgtaat tgtggcaact gacaatccta ctcacactcc acaagctgct tattatcaat 5340
tctacccacg ttagacaaac taccatgatt aacactgggt taagaaaaaa atagaatttt 5400
aagtgcagat atatcaataa atactgaaga accaccttaa aaatacataa atactgaaga 5460
acaggatttc tactggtgtt gctgtttttc ttttcttttt ttttaatctc caatcctatt 5520
tgtggcagat ttctggaaag tacaaattgt gttggtatgt gcacaaacct ttgcaagatt 5580
ccatgccaga agttcatcca agattcactt ggcatgaagg tctacatgtc tcccagtaag 5640
cttcctctgc tctgattcat caggaaagcc tgaatatcag tgggtaaaca caacaactta 5700
aatataattt ggtagaataa ggtaaaaata atcaaactcc attcctttgc ataaagttgg 5760
caagaaacta ataatggatt gtcatatcaa catcatctca aagttgaaaa tttgggttga 5820
aacctggtgt gcctctaact tctcaggaca ttgacctaat tttactatag cggatcatta 5880
tacggactgc cgctaaaaca taagtgaact caaaatatca actttcgact ttaagcagaa 5940
ttagtgaagt aaagttgaat caatgtaata ggtccataga ccaccaacgg tttagagcta 6000
tgagaaatac agaacttaat ttgacatatt aatttcatgc cacctctacc taccgacaat 6060
ctgaggactg tgcccttttt gggtatggga accactggcc accattcagg aaacagaaag 6120
ggctgaagtg gggccaacat gtttgcaggt tactacctcc gttttttaat ttataacgtc 6180
gatgactttt tagatatata tgatcattcg tcttattcaa aaaaaaatgc aattatcatt 6240
tattttattg tgacttaatt tatcatcaaa tgttctttaa gcatgactta aatttttttt 6300
atatttgcac aataattttg aataagatga atggtcaaat gtttgtcaaa aagtcaacaa 6360
cgtcatacat taaaaaacgg agggagtaca aagcaatcac aagcaagttg tacagaagaa 6420
gaaagtgcta gcccctactt gtcattctta gtgggaaacg gcaaattgca tggacaagtt 6480
gtggaacagt agtgtgaaat ccagaaatcc acaatgtagt gctgtgaaaa tacagtacaa 6540
gcaccttcac atggcagtga cgagccagca tgtctcgata atcagagcaa tatggttcaa 6600
tttcacttat gagaagcact agttttgaat gttagtcttc tatggtgata cactgaatat 6660
ttcaacatca atcatttttt ctaggtcaaa agatactgat attttatgac atcaccccac 6720
agaatcagat atcaacctct gttggaattt gcaaggctct ctttcctttt tatcctgtcg 6780
agtggttaga tctaggagta agcaatgaat gttatttact gccagtgcta ctaaatagtc 6840
ctatacagct tatttccaat taagcaagaa ggttttttcc aatactaggt caataaaaac 6900
gagaataaag cagagtccct aaagataata gtgaatgtat caagagaaat gtaacataat 6960
tacttacatc aactatatat ttattcttca gattttgaag acatgagctg tgagatgata 7020
tttggacagc aacctcctga agatgaccct gcattgaagc agccatgctt ccggacaaaa 7080
tgtaaggaat tccgcactga ggcaccttca tggattcaat caacactcct ctatattcat 7140
actttttact tcgtgtatat gcaggcgtcg caaagcagaa tcatggtgtg aattgctcca 7200
tctgatctga aagaaattat caatagatag atttcaaatc agtaaaatgc cttaagctcc 7260
atttccttta ttcctttgga aaaaaaatta gcaccatcat tgtttttgcc cacaacacca 7320
gcatgtttgg aacatacact cttcattgta atccaaaagt aatctaagag gaaatgaaag 7380
gcccaacagt aaccatttta aggtacatct actccatatg ttgttaaaag aggaattgaa 7440
gagcattgaa agcggggcat ggagttgccg atttcaaaaa gttctaatgt taatcagata 7500
aattaaatcg atgcacccct tgcagagaca tattctattt attactgtaa ttgtagaaaa 7560
tttcagttaa ttacagaaag atgtgagtgg aaaccaagta ctatttcatg gacgaaaatt 7620
ataatgcacg gattcctcta aaagaattta ttttaaaact ctaactgata taactctcga 7680
tccactatca gttattaggg gaaatattct atttgtcact aagtattttg aggaaaatag 7740
aattctatat atgtatatgg ctgtctatgc actgtctcat ccacccaatg tgctaaaaga 7800
tttgagcagg gacattgaaa ttattagaga tggcaatttt taaagtttgc atgactatcg 7860
aggcatatgg gacatatagg acagtgccta actcataaat atatgtcaga aaaccattct 7920
ccacactaca tcaggatatt agtatgaaat ggggaaactg aacatatcta catgtacttc 7980
tcatcagctt gcattttgta caaaaaaaaa accaaaacca agattgcaat gccagacctg 8040
cgggattttt aagttggaaa agaaaatgat tcagcacctc cagagaggtc catggttcta 8100
gcaaatacca aaagaaagta gctatctcac atattgattc tggagacatt ggtacaccaa 8160
ttatgacaga cttgaccaaa attgtataaa gtaacacaac tttaacatat tgtagaaaaa 8220
aaatagttgg gatctaccac actgatgaaa ccactttttg agacctagag cacttacagg 8280
ataaattcac ataaagaatt caaatgtcac aacaaaatca tgacagggat aaccattgct 8340
actgacaagg ggatcgtgcg ccacattact gttctttgtc tcctgatgca tgatcgtttc 8400
cgtagctcta ctggctttag tttattcttc acaagatttc cgcattattg ttctttgtct 8460
cctgatgcat gatctgtttt cataactcta ttgggtattg gctttagttg ttctccaaaa 8520
gattcagata aggtcctgac tgaatgtact acataactat agtctccatt aggattgctg 8580
gttcagtata acaaatggtg tacgcagaat tcaacaacca ttgagaaggc cagtttttat 8640
cgctagttcc tgaaattcta gaaatagtcc tgtattccaa atggggctaa aagtaaggaa 8700
attctatgtt gttaacaaag acaatagcct aaccccaagt aagttggagg ttagagatga 8760
aacccaagcc aagccacatc ctacatgaat gtcattacaa gtgcaaatct aatgaagctc 8820
caggttgtcc cagttttata ttgtacaaga taatctgcac gaatcatgat atttgattgt 8880
aaaaaggaaa acctaatact tcaaagtcag ttcaagaacg atgcaaatat gtgcagtact 8940
ttttgcaaat tagcttattg aaagaacgat ttaacaaata agaaacagga gaaacacaac 9000
attcctagct gttcactttt gcatgatatt caactaggag ccttgtgggg tggtggtgcc 9060
attttgcaag tgatacactt agtatggcac ctagctaatt tccaacctta tatacaataa 9120
tcaggtcaag gcctcactag gcttcaaaaa tgttagaacc tctgcagaaa tatcttacta 9180
tatattgata ctttaaaaaa ttgcaatctt gaaaagaaac tgtaaacaca aatgtatggt 9240
attggtacc 9249
<210>2
<211>837
<212>DNA
< 213>paddy rice (Oryza sativa L.)
<400>2
atggagacca ccacgcttgt gctgcttctt cctcatggcg gcgccggcgg cgtacggccg 60
gcggcagcgg caacggcgaa gcgaagctac gtgatgagga ggtgttgctc gacggtgagg 120
gcggtcatgg cgaggccgca agaggcgccg gcgtcggcgc cggccaagaa gacggagacg 180
gcggcgatga tgtcgacggt gcagacggag acggcggcgg cgccgccggc gacggtgtac 240
cgggacagct ggttcgacaa gctcgccatt ggttacctgt ccaggaacct tcaagaagct 300
tctgggctaa agaatgaaaa ggatggctac gagagcctga tagatgccgc cctagccatc 360
tcaagaatct tcagtctgga taaacaaagc gagattgtga cccaagctct tgaaagagca 420
cttccaagct acatcctcac aatgatcaag gtgatgatgc caccttcaag attttccagg 480
gagtactttg ctgcattcac cacgatattt tttccttggt tggttgggcc gtgtgaggtt 540
atggaatctg aagttgaagg aaggaaagag aaaaacgtgg tatatatccc caaatgcaga 600
tttctggaaa gtacaaattg tgttggtatg tgcacaaacc tttgcaagat tccatgccag 660
aagttcatcc aagattcact tggcatgaag gtctacatgt ctcccaattt tgaagacatg 720
agctgtgaga tgatatttgg acagcaacct cctgaagatg accctgcatt gaagcagcca 780
tgcttccgga caaaatgcgt cgcaaagcag aatcatggtg tgaattgctc catctga 837
<210>3
<211>833
<212>DNA
< 213>paddy rice (Oryza sativa L.)
<400>3
atggagacca ccacgcttgt gctgcttctt cctcatggcg gcgccggcgg cgtacggccg 60
gcggcagcgg caacggcgaa gcgaagctac gtgatgagga ggtgttgctc gacggtgagg 120
gcggtcatgg cgaggccgca agaggcgccg gcgtcggcgc cggccaagaa gacggagacg 180
gcggcgatga tgtcgacggt gcagacggag acggcggcgg cgccgccggc gacggtgtac 240
cgggacagct ggttcgacaa gctcgccatt ggttacctgt ccaggaacct tcaagaagct 300
tctgggctaa agaatgaaaa ggatggctac gagagcctga tagatgccgc cctagccatc 360
tcaagaatct tcagtctgga taaacaaagc gagattgtga cccaagctct tgaaagagca 420
cttccaagct acatcctcac aatgatcaag gtgatgatgc caccttcaag attttccagg 480
gagtactttg ctgcattcac cacgatattt tttccttggt tggttgggcc gtgtgaggtt 540
atggaatctg aagttgaagg aaagagaaaa acgtggtata tatccccaaa tgcagatttc 600
tggaaagtac aaattgtgtt ggtatgtgca caaacctttg caagattcca tgccagaagt 660
tcatccaaga ttcacttggc atgaaggtct acatgtctcc caattttgaa gacatgagct 720
gtgagatgat atttggacag caacctcctg aagatgaccc tgcattgaag cagccatgct 780
tccggacaaa atgcgtcgca aagcagaatc atggtgtgaa ttgctccatc tga 833
<210>4
<211>278
<212>PRT
< 213>paddy rice (Oryza sativa L.)
<400>4
Met Glu Thr Thr Thr Leu Val Leu Leu Leu Pro His Gly Gly Ala Gly
1 5 10 15
Gly Val Arg Pro Ala Ala Ala Ala Thr Ala Lys Arg Ser Tyr Val Met
20 25 30
Arg Arg Cys Cys Ser Thr Val Arg Ala Val Met Ala Arg Pro Gln Glu
35 40 45
Ala Pro Ala Ser Ala Pro Ala Lys Lys Thr Glu Thr Ala Ala Met Met
50 55 60
Ser Thr Val Gln Thr Glu Thr Ala Ala Ala Pro Pro Ala Thr Val Tyr
65 70 75 80
Arg Asp Ser Trp Phe Asp Lys Leu Ala Ile Gly Tyr Leu Ser Arg Asn
85 90 95
Leu Gln Glu Ala Ser Gly Leu Lys Asn Glu Lys Asp Gly Tyr Glu Ser
100 105 110
Leu Ile Asp Ala Ala Leu Ala Ile Ser Arg Ile Phe Ser Leu Asp Lys
115 120 125
Gln Ser Glu Ile Val Thr Gln Ala Leu Glu Arg Ala Leu Pro Ser Tyr
130 135 140
Ile Leu Thr Met Ile Lys Val Met Met Pro Pro Ser Arg Phe Ser Arg
145 150 155 160
Glu Tyr Phe Ala Ala Phe Thr Thr Ile Phe Phe Pro Trp Leu Val Gly
165 170 175
Pro Cys Glu Val Met Glu Ser Glu Val Glu Gly Arg Lys Glu Lys Asn
180 185 190
Val Val Tyr Ile Pro Lys Cys Arg Phe Leu Glu Ser Thr Asn Cys Val
195 200 205
Gly Met Cys Thr Asn Leu Cys Lys Ile Pro Cys Gln Lys Phe Ile Gln
210 215 220
Asp Ser Leu Gly Met Lys Val Tyr Met Ser Pro Asn Phe Glu Asp Met
225 230 235 240
Ser Cys Glu Met Ile Phe Gly Gln Gln Pro Pro Glu Asp Asp Pro Ala
245 250 255
Leu Lys Gln Pro Cys Phe Arg Thr Lys Cys Val Ala Lys Gln Asn His
260 265 270
Gly Val Asn Cys Ser Ile
275
<210>5
<211>227
<212>PRT
< 213>paddy rice (Oryza sativa L.)
<400>5
Met Glu Thr Thr Thr Leu Val Leu Leu Leu Pro His Gly Gly Ala Gly
1 5 10 15
Gly Val Arg Pro Ala Ala Ala Ala Thr Ala Lys Arg Ser Tyr Val Met
20 25 30
Arg Arg Cys Cys Ser Thr Val Arg Ala Val Met Ala Arg Pro Gln Glu
35 40 45
Ala Pro Ala Ser Ala Pro Ala Lys Lys Thr Glu Thr Ala Ala Met Met
50 55 60
Ser Thr Val Gln Thr Glu Thr Ala Ala Ala Pro Pro Ala Thr Val Tyr
65 70 75 80
Arg Asp Ser Trp Phe Asp Lys Leu Ala Ile Gly Tyr Leu Ser Arg Asn
85 90 95
Leu Gln Glu Ala Ser Gly Leu Lys Asn Glu Lys Asp Gly Tyr Glu Ser
100 105 110
Leu Ile Asp Ala Ala Leu Ala Ile Ser Arg Ile Phe Ser Leu Asp Lys
115 120 125
Gln Ser Glu Ile Val Thr Gln Ala Leu Glu Arg Ala Leu Pro Ser Tyr
130 135 140
Ile Leu Thr Met Ile Lys Val Met Met Pro Pro Ser Arg Phe Ser Arg
145 150 155 160
Glu Tyr Phe Ala Ala Phe Thr Thr Ile Phe Phe Pro Trp Leu Val Gly
165 170 175
Pro Cys Glu Val Met Glu Ser Glu Val Glu Gly Lys Arg Lys Thr Trp
180 185 190
Tyr Ile Ser Pro Asn Ala Asp Phe Trp Lys Val Gln Ile Val Leu Val
195 200 205
Cys Ala Gln Thr Phe Ala Arg Phe His Ala Arg Ser Ser Ser Lys Ile
210 215 220
His Leu Ala
225
<210>6
<211>22
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>6
atggagacca ccacgcttgt gc 22
<210>7
<211>21
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>7
tcagatggag caattcacac c 21
<210>8
<211>27
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>8
aagagctccc aagaagacgg agacggc 27
<210>9
<211>28
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>9
aaactagtca ccatgattct gctttgcg 28
<210>10
<211>27
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>10
aaggatcccc aagaagacgg agacggc 27
<210>11
<211>28
<212>DNA
< 213>artificial sequence
<220>
<223>
<400>11
aaggtaccca ccatgattct gctttgcg 28
Claims (13)
1. albumen, the protein of forming by the aminoacid sequence shown in the sequence in the sequence table 4.
2. the said proteic encoding sox of claim 1.
3. encoding sox according to claim 2 is characterized in that: said encoding sox is the dna molecular shown in the sequence 2 in the sequence table.
4. the recombinant vectors that contains claim 2 or 3 said encoding soxs.
5. the reorganization bacterium that contains claim 2 or 3 said encoding soxs.
6. the transgenic cell line that contains claim 2 or 3 said encoding soxs.
7. the expression cassette that contains claim 2 or 3 said encoding soxs.
8. the primer of amplification claim 2 or 3 said encoding sox total lengths is right.
9. the application of the described albumen of claim 1 in regulating rice tillering.
10. claim 2 or the 3 described encoding soxs application in regulating rice tillering.
11. the described albumen of claim 1 is the application in the gold lactone content of only angle in regulating paddy rice.
12. claim 2 or 3 described encoding soxs are the application in the gold lactone content of only angle in regulating paddy rice.
The method of the paddy rice of increasing is with claim that contains in the paddy rice 2 or 3 said encoding sox deactivations 13. a cultivation is tillered, and obtains the paddy rice that tillers and increase.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910083935XA CN101585869B (en) | 2009-05-11 | 2009-05-11 | Rice tillering associated protein, encoding gene and use thereof |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910083935XA CN101585869B (en) | 2009-05-11 | 2009-05-11 | Rice tillering associated protein, encoding gene and use thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN101585869A CN101585869A (en) | 2009-11-25 |
| CN101585869B true CN101585869B (en) | 2012-05-23 |
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| CN200910083935XA Expired - Fee Related CN101585869B (en) | 2009-05-11 | 2009-05-11 | Rice tillering associated protein, encoding gene and use thereof |
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| Country | Link |
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| CN (1) | CN101585869B (en) |
Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102731633B (en) * | 2011-04-01 | 2014-03-05 | 中国科学院遗传与发育生物学研究所 | Plant lateral branch number correlated transcription factor AtDOF 4.2, its encoding gene and application |
| CN112695042A (en) * | 2019-10-23 | 2021-04-23 | 华南农业大学 | ZmD53 application of gene in regulating development of maize tassel branches or breeding new variety of density-resistant plants |
| CN111378672A (en) * | 2020-03-17 | 2020-07-07 | 福建省农业科学院生物技术研究所 | Rice dwarf and multi-tillering gene Os11g0587000 mutant and application thereof |
| CN111848765B (en) * | 2020-07-22 | 2021-10-08 | 中国水稻研究所 | Rice gene OsFBK4 and mutant and application thereof |
| CN114539372A (en) * | 2021-04-06 | 2022-05-27 | 中国科学院遗传与发育生物学研究所 | Rice tillering angle control gene LAZY2 and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1477112A (en) * | 2002-08-20 | 2004-02-25 | 中国科学院遗传与发育生物学研究所 | Rice tiller control gene MOC1 and its application |
| CN1844396A (en) * | 2006-04-28 | 2006-10-11 | 中国农业大学 | Gene adjusting and controlling rice tillering angle and its coded protein and use |
-
2009
- 2009-05-11 CN CN200910083935XA patent/CN101585869B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1477112A (en) * | 2002-08-20 | 2004-02-25 | 中国科学院遗传与发育生物学研究所 | Rice tiller control gene MOC1 and its application |
| CN1844396A (en) * | 2006-04-28 | 2006-10-11 | 中国农业大学 | Gene adjusting and controlling rice tillering angle and its coded protein and use |
Non-Patent Citations (2)
| Title |
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| FUKUTA YOSHIMICHI et.al.Identification of low tiller gene in two rice varieties,Aikawa 1 and Shuho of rice(Oryza sativa L.).《JIRCAS Working Rep》.2006,第46卷86-92. * |
| HAO LIN et.al.DWARF27,an iron-containing protein required for the biosynthesis of strigolactones,regulates rice tiller bud outgrowth.《THE PLANT CELL》.2009,第21卷(第5期),15121525. * |
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| Publication number | Publication date |
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| CN101585869A (en) | 2009-11-25 |
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