CN106868021A - Control rice paddy seed size gene OsNAC1 and its application - Google Patents

Abstract

The present invention provides a kind of control rice paddy seed size geneOsNAC1, from paddy rice (Oryza sativaL. 11) are spent in cv., its nucleotides sequence is classified as SEQ ID NO.1, the nucleotides sequence of its gene open reading frame is classified as SEQ ID NO.2, and the amino acid sequence of its cDNA codings is SEQ ID NO.3.Overexpressed in paddy riceOsNAC1After find, compared with wild rice,OsNAC1Overexpression transgenic paddy rice seed diminishes, and seed width and seed length are all reduced, and mass of 1000 kernel reduces, and plant height reduction, setting percentage is constant.These result explanationsOsNAC1It is the gene for controlling rice paddy seed size.Additionally, we have planted 4 generation transgenic lines, and since the reduction of T1 seed size and mass of 1000 kernel all conspicuousnesses for, illustrate what we obtainedOsNAC1Transfer-gen plant can be stabilization heredity.

Description

Control rice paddy seed size gene OsNAC1 and its application

Technical field

The invention belongs to plant biotechnology field, it is related to a kind of size of plant seed indispensable gene and its application, tool Body is related to a kind of control rice paddy seed size geneOsNAC1And its application.

Background technology

Paddy rice is the important cereal crops of China, in face of the severe shape of population increase, environmental degradation and Cultivated Land Area Decrease Gesture, it is ensured that grain security is as one that China the faces big problem and severe challenge for becoming increasingly conspicuous, and it is main to improve paddy rice etc. The yield per unit area of cereal crops is the important channel for solving this problem, to ensureing the grain security of China, promoting its people Expanding economy plays the role of very important.Number of productive ear, number of grain per ear and mass of 1000 kernel are three of composition rice yield basic Key element.Seed size is by grain length, grain is wide, grain is thick and grain plumpness is together decided on, be the topmost decision of mass of 1000 kernel because Element.Seed size is also one of Other Main Agronomic Characters of composition Ideal Rice Plant Type, for a long time always many crop breedings The important goal of improvement, has a great impact to yield and exterior quality.The utilization of " hybrid vigour " and " green revolution ", makes me State's rice yield realizes two forward leaps.Hybrid rice is mainly based upon the discovery of wild abortion type sterile series and the hair of three series mating method It is bright, yield is substantially improved." green revolution " is then based on Semi dwarfism genesd1Discovery and utilization, plant height is significantly dropped It is low, resistance to fertilizer and lodging tolerance are improve, so as to significantly improve yield (Sasaki et al., 2002).During the nearly last ten years, " super hybridization rice " breeding enjoys people to expect, its topmost technology for using is exactly that ideotype and use of advantage are combined.By This is visible, and the breakthrough in Ideal Rice Plant Type research is the premise and basis that rice breeding can more rapid and better be carried out down, Therefore, the excavation for one of the important factor that is built up as ideotype seed size correlation function gene and its mechanism of action Research, and then the cultivation of the cereal crop high-yield variety such as paddy rice, wheat is applied to, with important theory significance and production Application prospect.

Grain high yield, high-quality are the important goals of plant basic science and applied science research, are also cereal crops heredity The most important thing of improvement.Paddy rice is important cereal crops and model organism, in the cereal crops such as paddy rice, wheat, seed Size is both paddy rice Appearance Quality Traits, is again the target proterties of a very important yield traits and Crop Domestication.In recent years Clone and functional study to arabidopsis and rice paddy seed size related gene show that Ubiquitin-proteasome is probably a kind of regulation and control The mechanism of seed size.GW2Gene code one has the RING albumen of E3 ubiquitin ligase activities,GW2Afunction is mutated The number of rice cell is increased, causes glume to become big, filling rate is accelerated, so that grain is wide and grain increases again, output increased, Result shows that GW2 is degraded by the way that its substrate is anchored into proteasome, so that the division of negative regulator cell（Song et al., 2007）.Additionally, knocking out wheatTaGW2Gene makes wheat seed diminish, and grain is reduced again（Su et al., 2011）.Water RiceGW5One nuclear locating sequence of 144 amino acid composition of gene code, the albumen includes a nuclear localization signal and a richness Region containing arginine.Confirm that GW5 has interaction with poly ubiquitin by yeast two-hybrid assay, show that GW5 may be by general Grain is wide and grain weight for the regulation of fibroin enzyme body approach.gw5Ubiquitin can not be transferred on target protein after mutation, so that should drop The substrate of solution can not be by specific recognition, and then activation grain husk spends the division of outer hülle cell, so that increase the width of grain husk flower shell, finally The width of husk, grain are heavy and yield is obtained for increase（Song et al., 2011）.SOD2Gene code one removes ubiquitin Change enzyme UBP15 (ubiquitin-specific protease 15), overexpressionSOD2Gene causes arabidopsis seed to become big, Andsod2Single mutant reduces arabidopsis seed size, shows that SOD2/UBP15 is the key factor of seed size regulation and control, carefully Born of the same parents learn analysis showsSOD2It is by promoting the cell division of integument, so as to influence seed size.The experiment such as co-immunoprecipitation is taken off Show that ubiquitin acceptor DA1 can show that SOD2 is the direct substrate of DA1 with SOD2 in vivo and external directly interaction.Tie above Fruit shows：Ubiquitin acceptor DA1 by directly with deubiquitinating enzymes SOD2 interactions, and the degraded of SOD2 is mediated, so as to regulate and control plant Seed size（Du et al., 2014）.Current research finds that E3 ubiquitin ligases DA2 and EOD1 are swashed by ubiquitination DA1 Live the proteinase activity of DA1, but the DA1 of activation can further cut the stream substrates such as UBP15, TCP15, TCP22, enter And regulate and control seed size, show that the proteinase activity of DA1 regulates and controls seed size by the stability of coordinated regulation stream substrates （Dong et al., 2017）.

In recent years, it is found that some IKU pathway genes participate in seed size regulation and control in plant（Fig. 1）.IKU1Gene is compiled One albumen comprising VQ motifs of code, expresses in early stage endosperm,IKU1Afunction mutation reduces arabidopsis seed size （Wang et al., 2010）.IKU2One receptor kinase rich in leucine of coding,IKU2Afunction mutation is also reduced Seed size（Li et al., 2016）.SHB1Its crucial regulating and controlling effect in seed development, overexpresses in mustard and intends south MustardSHB1Gene increased Mustard seed size and yield（Savadi et al., 2015）.Recent studies have found that,SHB1Can With reference toMIN3WithIKU2Gene promoter region, by changingMIN3WithIKU2The expression of gene regulates and controls the development of endosperm, ButABI5Directly withSHB1Promoter is combined and then suppressedSHB1Expression, rape is overexpressed in arabidopsisIKU2Gene increases Seed size is added（Li et al., 2016; Xiao et al., 2016）.abi5Mutant increased seed size, knot Fruit show ABA regulation and control seed development be at least partly byABI5MediationSHB1Caused by the suppression of gene expression（Cheng et al., 2014）.

Rice paddy seed size is a gene expression regulation process for complexity, and seed size is by the quantity of controlled by multiple genes Proterties.Research advantage on its morphology of paddy rice and science of heredity is non-for the molecular mechanism research of higher plant seed size is provided Often important research mode.Therefore, excavate new control rice paddy seed size gene, to cereal crop molecular breeding and high yield The cultivation of kind has important application value and theory significance.

The content of the invention

It is an object of the invention to provide one kind control rice paddy seed size geneOsNAC1And its application, the gene is water The key controlling gene of rice size, it can be used for the cultivation of the cereal crop high-yield variety such as paddy rice, wheat.

The purpose of the present invention is realized in the following manner：

The present invention provides a kind of control rice paddy seed size geneOsNAC1, from paddy rice (Oryza sativa L.) cv. In spend 11, its nucleotides sequence is classified as SEQ ID NO.1, and the nucleotides sequence of its gene open reading frame is classified as SEQ ID NO.2, The amino acid sequence of its cDNA codings is SEQ ID NO.3.

A kind of recombinant precursor, including control rice paddy seed size geneOsNAC1Nucleotide sequence, the construct Carrier is the pMD18-T carriers for cloning or the pCAMBIA1301 carriers for expressing.

Control rice paddy seed size geneOsNAC1Nucleotide sequence transgenic Agrobacterium be EHA105.

Control rice paddy seed size geneOsNAC1Nucleotides sequence be listed in plant tissue expression in application.

The plant tissue is root, stem, leaf, flower, fringe or seed.

The plant is monocotyledon or dicotyledon.

The monocotyledon be paddy rice, wheat, barley, sorghum or corn, the dicotyledon be arabidopsis, kind Eggplant, tobacco, soybean or potato.

Above-mentioned control rice paddy seed size geneOsNAC1Or above-mentioned control rice paddy seed size geneOsNAC1Coding Function albuminoid in protein, or other species, its amino acid sequence and the amino acid sequence tool shown in SEQ ID No.3 There is the homology not less than 30%, application of the albumen in the plant that breeds high-yield variety.The high yield is increase cereal crop Seed size, and then increase per unit area yield.

Relative to prior art, the advantage of the invention is that：

It is right firstOsNAC1Gene has carried out tissue specific expression analysis, and qRT-PCR results showOsNAC1 Gene is in water Rice spire high efficient expression；Then willOsNAC1It is gene constructed to the plant expression vector pCAMBIA1301 started with 35S promoter In, obtain recombinant vector pCAMBIA1301-OsNAC1, recombinant vector is transferred to Agrobacterium for further converting.With paddy rice Callus is acceptor, and the carrier that will be built using agrobacterium-mediated transformation is transferred to paddy rice, is dyeed by hygromycin and GUS and is turned Gene strain.Overexpressed in paddy riceOsNAC1After find, compared with wild rice,OsNAC1Overexpression transgenic paddy rice kind Son diminishes, and seed width and seed length are all reduced, and mass of 1000 kernel reduces, and plant height reduction, setting percentage is constant.These result explanationsOsNAC1It is the gene for controlling rice paddy seed size.Additionally, we have planted 4 generation transgenic lines, and opened from T1 generations The reduction of beginning seed size and mass of 1000 kernel all conspicuousnesses, illustrates what we obtainedOsNAC1Transfer-gen plant can be stabilization heredity 's.OsNAC1Gene is the key controlling gene of rice paddy seed size, shows that technique for gene engineering regulation paddy rice can be utilized Seed size, purposefully regulating and controlling plant characters with plant using biotechnology, adjustment plant population's structure with reach high yield, High-quality aspect has very important application value.The gene can be used for the standing grain such as paddy rice, wheat by CRISPR/Cas9 technologies The cultivation of cereal crops high-yield variety.

Brief description of the drawings

Fig. 1 represents expression quantity of the OsNAC1 in wild rice different tissues.

Fig. 2 is expression quantity detection of the OsNAC1 genes in wild type and transgenic line.

Fig. 3 is the seed size phenotype of OsNAC1 genes overexpression plant and adjoining tree.

Fig. 4 is OsNAC1 genes overexpression plant and adjoining tree mass of 1000 kernel.

Fig. 5 is the phenotype of OsNAC1 genes overexpression plant and adjoining tree.

Fig. 6 is OsNAC1 genes overexpression plant and adjoining tree plant height.

Fig. 7 is OsNAC1 genes overexpression plant and each panel length statistical analysis of adjoining tree.

Fig. 8 is OsNAC1 genes overexpression 20 seed length phenotypes of plant and adjoining tree.

Fig. 9 is OsNAC1 genes overexpression plant and adjoining tree seed length statistical analysis.

Figure 10 is OsNAC1 genes overexpression 20 seed width phenotypes of plant and adjoining tree.

Figure 11 is OsNAC1 genes overexpression plant and adjoining tree seed width statistical analysis.

Figure 12 is OsNAC1 genes overexpression plant and adjoining tree setting percentage statistical analysis.

Specific embodiment

It is further right that present invention below is intendedOsNAC1The upstream and downstream gene of gene and its associated regulatory signal path of participation It is analyzed, and analyzesOsNAC1Whether gene has the effect of widely control seed size.Existed by analyzing the gene Effect in rice paddy seed size control study mechanism and high-yield variety cultivation is big with the cereal crop seed such as horn of plenty paddy rice Small molecule mechanism stockpile, is set by the high yield molecular modules that genetic engineering means are the cereal crops such as paddy rice, wheat Meter breeding provides new genetic resources.

Transgenic Rice strain is obtained and phenotypic analysis experiment

1 materials and methods

1.1 vegetable materials and planting patterns

For the rice varieties that try for spend in paddy rice japonica rice variety 11 i.e. paddy rice (Oryza sativaL. 11) are spent in cv., is protected There is Zhoukou Normal University's plant genetic and molecular breeding key lab.It is first dense with mass percent before the new seed plantation received Spend for the min of 5% hypochlorite disinfectant 40 or sterilized 12 h with 1/1000 carbendazim, then with 0.1 mol/L HNO₃（0.1 mol/L HNO₃Compound method：The dense HNO of 1 mL 63%₃Add 100 mL water）Soak seed 16 h, and running water will after rinsing well Seed after sterilization is placed under 28 °C of environment 1 d that soaks seed, and is then uniformly laid in culture dish seed, wherein in culture dish A metafiltration paper is placed, and is moistened with water；Close the lid, cultivated under being then placed in 32 °C, water is changed daily two to three times, it is seen that mostly Several sub- broken shells are cultivated under going out after root to be transferred to 30 °C.The 2d that first basked seeds before seed more than half a year is sowed is preserved, is soaked seed after sterilization, disappeared Malicious method：55 °C of 30min that hot water treatment of seeds, treat rice bud grow to 4.8-5.2 mm it is long when be seeded on window screen cloth, and in water Cultivated in rice nutrient solution, after 3 leaf phases, moving into soil and carry out corresponding label rice seedling carries out Phenotypic Observation and follow-up Analysis of experiments.Paddy rice carries out single-dose application at interval of 8 or 10d.

1.2 agents useful for same and carrier

The Escherichia coli that this experiment is used are DH5 α, and Agrobacterium is EHA105, and these bacterial strains are that Zhoukou Normal University plant is lost Pass and preserved with molecular breeding key lab, plant expression vector is pCAMBIA1301；Wherein, restriction enzyme, Ke Longzai Body pMD18-T, T4 DNA ligase, Taq archaeal dna polymerases are purchased from TaKaBa biotech firms；DNA QIAquick Gel Extraction Kits are Magen Biotech firm's product；Hygromycin（Hyg）, kanamycins（Kan）With ammonia Ka-7038Ⅶ（Amp）Have Deng purchased from Beijing ancient cooking vessel state biotechnology Limit company, tests all primers and synthesizes by the prosperous biotech firm of Beijing Losec.

The extraction of 1.3 RNA, cDNA synthesize and RT-PCR amplifying target genes

RNA extracts the plant RNA extraction kit using Magen companies, and reference method is to be easier to extract plant tissue RNA in a small amount Extraction method.Template is done with the RNA of 1 μ g, according to cDNA synthetic agent box（TaKaRa）Operating instruction synthesizes the first chain cDNA.According toOsNAC1Gene cDNA full length sequence designs special primer, and specific primer sequences are shown in Table 1.RT-PCR reaction systems （20 μL）：10 × PCR reaction buffers 2 μ L, dNTP（2.5 mmol/L）1 μ L, primer（10 pm/μL）Each 1 μ L, Taq Polymerase（5 U/µL）0.2 μ L, template cDNA 2 μ L, ddH₂O 12.8 µL.PCR reaction conditions：94 DEG C of predegenerations 5 min；94 DEG C of denaturation 30 s, 54 DEG C of annealing 30 s, 72 DEG C of 1 min of extension, 33 circulations；72 DEG C of 10 min of extension, 4 DEG C of guarantors Deposit.

1.4 OsNAC1Tissue-specific expression is analyzed

OsNAC1Real-time fluorescence quantitative PCR is selected in gene expression amount detection（qRT-PCR）.QRT-PCR results show：OsNAC1 Highest is expressed in spire, as shown in Figure 1.For qRT-PCR analysis primer according to full length gene sequences Design, from paddy rice 'sUbiquitin1(Ubi1;Os06g0681400) as quantitative PCR internal control primer, as shown in table 2.

1.5 OsNAC1The structure of gene plant expression vector

The PCR primer of genes of interest, is purified according to the PCR primer purification kit operation of Megan companies.Restriction enzymeKpnI andXbaAfter I double digestions are connected with the pMD18-T plasmids of genes of interest fragment or pCAMBIA1301 plasmids, T is used₄DNA connects Connect enzyme genes of interest is connected on plant expression vector pCAMBIA1301, reaction system is as follows：T₄DNA ligase（5 U/µ L）1 μ L, 10 × buffer 1 μ L, the μ L of 5 μ L, pCAMBIA1301 carrier of genes of interest fragment 3；Reaction condition：16 DEG C, 2 h. Connection product transformed competence colibacillusE.coliDH5 α, are coated with LB flat boards（50 mg/L Kan）Culture, 37 DEG C are overnight inverted culture shape Into single bacterium colony.

1.6 OsNAC1The identification of gene plant expression vector

PickingOsNAC1Gene plant expression vector plasmid is convertedE.coliThe monoclonal formed after DH5 α, extracting plasmid is carried out PCR is identified.Positive plasmid is converted to competence Agrobacterium EAH105, is coated with LB flat boards（50mg/L Kan、50mg/L Rif）Training Support, 28 DEG C are inverted culture 2d, select positive colony and extract plasmid and carry out digestion verification.

1.7 Agrobacteriums infect the acquisition with transgenic seedling

With Rice Callus as experiment material.OsNAC1Gene plant expression vector（Plasmid）Agrobacterium is converted by freeze-thaw method EHA105.Picking contains respectivelyOsNAC1Gene plant overexpression vector Agrobacterium monoclonal takes 10 mL in 28 DEG C of overnight incubations Nutrient solution, 3000 rpm, 20 min, is collected by centrifugation bacterial sediment, is resuspended in respectively in AAI nutrient solutions, and AA nutrient solutions include 30 G/L sucrose, 70g/L glucose, 200 μm of ol/L acetosyringones, PH 5.2, OD₆₀₀=1.0, then by suspension in shaking table Upper 28 DEG C of shaken cultivation 3-5 h.The Rice Callus that a certain size will be grown to are chosen, and are put into agrobacterium suspension and contaminate 30 min；Then callus is taken out, is placed in sterilizing filter paper and drains 50 min；Callus is placed in co-cultivation base（Co-culture Base is：2N6,10g/L glucose, 200 μm of ol/L acetosyringones, PH 5.5）On, 28 DEG C of d of light culture 3.Used after three days The aqua sterilisa of the U.S. element of 500 mg/L cephalos is cleaned 6 times, and the 100mL aqua sterilisas comprising 16 μ L tweens are cleaned 5 times, then use sterilized water Cleaning one time.The rice callus that will be drained are transferred to screening and culturing medium（Screening and culturing medium is：2N6, the U.S. element of 500 mg/L cephalos, 50 mg/L hygromycin）, 28 DEG C of d of light culture 30.The resistant calli that will newly grow is transferred to differential medium（Differential medium For：The mg/L6-BA of+30 g/L sorbierites of MS+30 g/L sucrose+2（The dance KT of flower）The mg/L NAA+ of+0.8% agar+1.0 The g/L caseinhydrolysates of+50 mg/L hygromycin of 250 mg/L cephalosporins+2, PH 5.8）.There are the rice callus of green bud in picking Move into and root media is housed（Root media is：The mg/L tides of+250 mg/L cephalosporins of MS/2+30g/L sucrose+50 are mould Element）Triangular flask in, be put into 28 DEG C of optical culture 15d of constant incubator.Prepare to transplant.First by hygromycin gene and GUS activity analysis carries out preliminary screening to positive transgenic plant, then by qRT-PCR technology for detection genes of interest wild Expression in type and transfer-gen plant, is further screened again to the first positive plant for screening.

1.8 OsNAC1Expression detection of the gene in wild type and transgenic paddy rice

The wild type and transgenic paddy rice blade for taking 14 d carry out RNA extractions, and template is done with the RNA of 1 μ g, are closed according to cDNA Into kit（TaKaRa）Operating instruction synthesizes the first chain cDNA.WithOsNAC1Gene cDNA designs special quantification PCR primer, 2 are shown in Table, are detected by qRT-PCROsNAC1Expression in wild type and transgenic line.

1.9 OsNAC1Phenotypic analysis of the gene in wild type and transgenic paddy rice

Be have detected by qRT-PCROsNAC1Gene is in wild type and transgenic lineOvOsNAC1（OE1, OE2 and OE3）In Expression, as shown in Figure 2；OverexpressionOsNAC1The seed size for reducing paddy rice of gene conspicuousness, as shown in figure 3, super ExpressionOsNAC1The reduction of plant mass of 1000 kernel conspicuousness, as shown in figure 4, plant height reduction is as shown in Fig. 5,6 and 7；With wild type phase Than overexpressionOsNAC1The reduction of plant seed length conspicuousness, as shown in FIG. 8 and 9, overexpressionOsNAC1Plant seed width shows Work property is reduced, and as shown in FIG. 10 and 11, setting percentage is constant, as shown in figure 12.The result is further to makeOsNAC1Gene application Had laid a good foundation and new genetic resources in paddy rice, High-yield Wheat Varieties improvement.

2.0 OsNAC1Sequence pair answers the total length of gene

OsNAC1It is 2292bp that sequence pair answers the full length sequence of gene, wherein be 1 089bp comprising complete reading frame sequence length, The following SEQ ID NO.1 of sequence；

NCBI ORF-finder are used to predict that its coded protein is 362 amino acid, sequence is as shown in SEQ ID NO.3.

According to sequence analysis,OsNAC1It is the new gene related to rice paddy seed size that One function is unknown, does not have still Gene known to any function is found in having the gramineous crop homologous with it.

Above example is only used to illustrate, the technical scheme being not intended to limit the present invention, although with reference to above-described embodiment to this Invention has been described in detail, it will be understood by those within the art that：The present invention can still be modified or Equivalent, any modification or partial replacement without departing from the spirit and scope of the present invention, it all should cover of the invention In the middle of right.

SEQUENCE LISTING

<110>Zhoukou Normal University

<120>Control rice paddy seed size gene OsNAC1 and its application

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<160> 3

<170> PatentIn version 3.3

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<213>The nucleotide sequence of control rice paddy seed size gene OsNAC1

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gaaaggccaa tatctcaaga gagctagcta gagctccatt tacaaccatt acaagattgc 60

atcagagaag atatcttcat agataattgt tggagcaaga acacaagaaa acacgtaggt 120

tcaattataa tcgatcgtgg agatggtgac cagcaaggag tttgcaaggg atcaggcggc 180

catggatcag aagatcaaga gcgacgtcgg cgaggtggtg ctcgccggag acgaggaaga 240

agacggggac gtggtgctcc cggggttcag gttccacccg accgacgagg agctcgtcac 300

gttctacctc cggcggaagg tggcgaggaa gtctctcagt atagagatca tcaaggagat 360

ggacatctac aagcacgatc catgggatct ccccagtaag tgcatcccca attaatttga 420

caaataacct caaacacaaa tcattttcaa aataattgag ctaaattctg tgtttgtgtt 480

tgaatgaatg gaatgatttt gtgcagatgc gagcacggta ggtggagaga aggagtggta 540

cttcttctgc ctgagaggga ggaagtacag gaacagcatc aggcccaaca gggtgacggg 600

atccgggttc tggaaggcga cggggatcga ccggccgatc tactccgccg ccgtcaacag 660

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caagggcacc aagaccgact ggatgatgca cgagttccgc ctcccgccgg cgatcgccgc 780

cgccgacgcc tccccgtgca tgcaagaagc tgtaagcacg cactcccatg ctctcatgct 840

cgttcgttct ccttctccat cgtcgtcaag gcctcgtgta ctcgatccca tgtacgtaag 900

aagtgcattc gtaatgggag aattacccaa actcagcctg gagagagatg gagaattacc 960

cagactcaac ctggacgaga gagatgggtc agactagagg gatacgagca accaaacaca 1020

ccatatccat atccggttct cctcatctgt gacagatcac aactacgtac aatttgatag 1080

tgtggagtac aatatcatct gcaatacttt acatcaccaa atcgtcttaa tatttttttt 1140

caagctaagt tcagcagttc tttcgagatt acaggcctcc ttttgaacga agaaattttg 1200

cgaaaattct acaaaaattg aactaatcca tgtgaaagtc ctactgaaat tcccacgttt 1260

taaaggagta ctggacatcc tctcacgttc tgacgtgttt ctagtattct accgattgcc 1320

atttctaacc gataagagta cagcctccta atagcattga cctctaactt gtgcacttct 1380

gactgaaaaa aatgcaggag gtctggacca tctgcagaat cttcaagaga agcatcacct 1440

acaggaagca gcagcagcag caggcctgga ggccgccggc gacggtgacc gtcaaggccc 1500

caccgccggg ggactcgagc tccaacaccg gcagcttcga gtcggacggc ggcggcgacg 1560

agttcatgaa ctgcggcctg actccggcca tctcccagca gcagcagcac ggcggccgtc 1620

atcagatgat gagcacgatg agctgcaacg gcggctactt cttcaacgac ggcatccatc 1680

acagccacag ccaccacaag cttcatagtc aatggggctc cctacaaatg gcgccgccgg 1740

agccgaagcc ggagccggag cagaagcctc tgagctcgcc ggcgatgacg atcgccttcc 1800

atcaaaacga ccatggcttc cccgccgccg ccgcggattt ctacaaggat gggtacttgg 1860

aagagattgc gaggatgatg gaggtggctg atccaagtcc aacaggattc tatgactgta 1920

gatattgatc agtaggaaca gggtttcaga atagatgcta gatatgcaga attcgccgac 1980

aagaacagtg ccccctttgt gtgagaggct gacatgtagg gcccatgctg cagaggctgc 2040

tatcactatt agtcttgtac ttttgttttc tgatctaata tatacactcc agatgggatg 2100

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ggctatatgc atactacacc tgtttcaaaa tataagtgat attggctata caattcatag 2220

ataatatctc ttagattttg agatggaggt aatatctctt agtacttaat gaatcatttt 2280

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<213>The gene open reading frame of the nucleotide sequence of control rice paddy seed size gene OsNAC1

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atggtgacca gcaaggagtt tgcaagggat caggcggcca tggatcagaa gatcaagagc 60

gacgtcggcg aggtggtgct cgccggagac gaggaagaag acggggacgt ggtgctcccg 120

gggttcaggt tccacccgac cgacgaggag ctcgtcacgt tctacctccg gcggaaggtg 180

gcgaggaagt ctctcagtat agagatcatc aaggagatgg acatctacaa gcacgatcca 240

tgggatctcc ccaatgcgag cacggtaggt ggagagaagg agtggtactt cttctgcctg 300

agagggagga agtacaggaa cagcatcagg cccaacaggg tgacgggatc cgggttctgg 360

aaggcgacgg ggatcgaccg gccgatctac tccgccgccg tcaacagcaa ctccggcgag 420

tcgatcgggc tgaagaagtc gctcgtctac taccgcggca gcgccggcaa gggcaccaag 480

accgactgga tgatgcacga gttccgcctc ccgccggcga tcgccgccgc cgacgcctcc 540

ccgtgcatgc aagaagctga ggtctggacc atctgcagaa tcttcaagag aagcatcacc 600

tacaggaagc agcagcagca gcaggcctgg aggccgccgg cgacggtgac cgtcaaggcc 660

ccaccgccgg gggactcgag ctccaacacc ggcagcttcg agtcggacgg cggcggcgac 720

gagttcatga actgcggcct gactccggcc atctcccagc agcagcagca cggcggccgt 780

catcagatga tgagcacgat gagctgcaac ggcggctact tcttcaacga cggcatccat 840

cacagccaca gccaccacaa gcttcatagt caatggggct ccctacaaat ggcgccgccg 900

gagccgaagc cggagccgga gcagaagcct ctgagctcgc cggcgatgac gatcgccttc 960

catcaaaacg accatggctt ccccgccgcc gccgcggatt tctacaagga tgggtacttg 1020

gaagagattg cgaggatgat ggaggtggct gatccaagtc caacaggatt ctatgactgt 1080

agatattga 1089

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<212> PRT

<213>The nucleotide sequence coded protein of control rice paddy seed size gene OsNAC1

<400> 3

Met Val Thr Ser Lys Glu Phe Ala Arg Asp Gln Ala Ala Met Asp Gln

1 5 10 15

Lys Ile Lys Ser Asp Val Gly Glu Val Val Leu Ala Gly Asp Glu Glu

20 25 30

Glu Asp Gly Asp Val Val Leu Pro Gly Phe Arg Phe His Pro Thr Asp

35 40 45

Glu Glu Leu Val Thr Phe Tyr Leu Arg Arg Lys Val Ala Arg Lys Ser

50 55 60

Leu Ser Ile Glu Ile Ile Lys Glu Met Asp Ile Tyr Lys His Asp Pro

65 70 75 80

Trp Asp Leu Pro Asn Ala Ser Thr Val Gly Gly Glu Lys Glu Trp Tyr

85 90 95

Phe Phe Cys Leu Arg Gly Arg Lys Tyr Arg Asn Ser Ile Arg Pro Asn

100 105 110

Arg Val Thr Gly Ser Gly Phe Trp Lys Ala Thr Gly Ile Asp Arg Pro

115 120 125

Ile Tyr Ser Ala Ala Val Asn Ser Asn Ser Gly Glu Ser Ile Gly Leu

130 135 140

Lys Lys Ser Leu Val Tyr Tyr Arg Gly Ser Ala Gly Lys Gly Thr Lys

145 150 155 160

Thr Asp Trp Met Met His Glu Phe Arg Leu Pro Pro Ala Ile Ala Ala

165 170 175

Ala Asp Ala Ser Pro Cys Met Gln Glu Ala Glu Val Trp Thr Ile Cys

180 185 190

Arg Ile Phe Lys Arg Ser Ile Thr Tyr Arg Lys Gln Gln Gln Gln Gln

195 200 205

Ala Trp Arg Pro Pro Ala Thr Val Thr Val Lys Ala Pro Pro Pro Gly

210 215 220

Asp Ser Ser Ser Asn Thr Gly Ser Phe Glu Ser Asp Gly Gly Gly Asp

225 230 235 240

Glu Phe Met Asn Cys Gly Leu Thr Pro Ala Ile Ser Gln Gln Gln Gln

245 250 255

His Gly Gly Arg His Gln Met Met Ser Thr Met Ser Cys Asn Gly Gly

260 265 270

Tyr Phe Phe Asn Asp Gly Ile His His Ser His Ser His His Lys Leu

275 280 285

His Ser Gln Trp Gly Ser Leu Gln Met Ala Pro Pro Glu Pro Lys Pro

290 295 300

Glu Pro Glu Gln Lys Pro Leu Ser Ser Pro Ala Met Thr Ile Ala Phe

305 310 315 320

His Gln Asn Asp His Gly Phe Pro Ala Ala Ala Ala Asp Phe Tyr Lys

325 330 335

Asp Gly Tyr Leu Glu Glu Ile Ala Arg Met Met Glu Val Ala Asp Pro

340 345 350

Ser Pro Thr Gly Phe Tyr Asp Cys Arg Tyr

355 360

Claims (8)

1. it is a kind of to control rice paddy seed size geneOsNAC1, its nucleotides sequence is classified as SEQ ID NO.1, and its gene open is read The nucleotides sequence of frame is classified as SEQ ID NO.2, and the amino acid sequence of its cDNA codings is SEQ ID NO.3.

2. a kind of recombinant precursor, it is characterised in that the nucleotide sequence of the gene including claim 1, the construct Carrier is the pMD18-T carriers for cloning or the pCAMBIA1301 carriers for expressing.

3. the transgenic Agrobacterium of the nucleotide sequence of gene as claimed in claim 1 is EHA105.

4. the nucleotides sequence of gene as claimed in claim 1 is listed in the application in plant tissue expression.

5. the nucleotides sequence of gene as claimed in claim 4 is listed in the application in plant tissue expression, it is characterised in that：It is described Plant tissue is root, stem, leaf, flower, fringe or seed.

6. the nucleotides sequence of gene as claimed in claim 4 is listed in the application in plant tissue expression, it is characterised in that：It is described Plant is monocotyledon or dicotyledon.

7. the nucleotides sequence of gene as claimed in claim 6 is listed in the application in plant tissue expression, it is characterised in that：It is described Monocotyledon be paddy rice, wheat, barley, sorghum or corn, the dicotyledon be arabidopsis, tomato, tobacco, soybean or Potato.

8. a kind of function albuminoid, it is characterised in that：Its amino acid sequence and the amino acid sequence shown in SEQ ID No.3 Row have the homology not less than 30%, application of the albumen in the plant that breeds high-yield variety.