CN109666672A - For reducing the reagent set and method of plant plant height - Google Patents

Abstract

The invention discloses the reagent sets and method for reducing plant plant height.Reagent set disclosed by the invention for reducing plant plant height is made of the promoter of entitled STK and the protein of entitled bin2-1；STK is DNA molecular shown in 1-3093 of sequence 1 in sequence table；Bin2-1 is the protein that amino acid sequence is sequence 2.It is demonstrated experimentally that can not influence leaf area while reducing plant plant height using reagent set of the invention, i.e., while not changing leaf blade size to reach the supply for guaranteeing plant nutrition, the plant height for reducing plant prevents plant from lodging.

Description

For reducing the reagent set and method of plant plant height

Technical field

The present invention relates in field of biotechnology, for reducing the reagent set and method of plant plant height.

Background technique

Tradition research shows the nutrient growth of plant and reproductive growth is inseparable, between each organ of plant life Long there is the relationships for interdepending, mutually promoting, and here it is the correlations of plant growth.The main body of the correlation of plant growth Present root and 3 aerial part, stem and side shoot, nutrient growth and reproductive growth aspects.Nutrient growth is the base of reproductive growth Plinth and premise, under normal operation, the vigorous leaf area of nutrient growth is big, photosynthate is more, and fruit and seed could well develop； Conversely, nutrient growth is bad to will lead to that plant is short and small thin and weak, and development of floral organs is incomplete, and fruit development is slow, and fruit is small, kind Subnumber mesh is few, low output.All material bases of plant reproductive growth are built upon on the basis of nutrient growth, nutrition organs Quality will have a direct impact on the development of reproductive organs, be not contemplated that one plant of thin short plant can the big seed of fringe it is more, full seed.? In real life, the plant that often encountering has generates the character easily to lodge since plant height is higher, to will lead to yield very It is low even without yield, in this case, in order to make plant is resistant to lodging to need to reduce plant plant height, but the reduction meeting of plant plant height It along with becoming smaller for nutrition organs (such as blade), and then may result in the decline of seed production, therefore, how to reduce plant The growth for not influencing nutrition organs while plant height is current urgent problem to be solved.

Summary of the invention

The technical problem to be solved by the present invention is to how not influence while reducing plant plant height the leaf area of plant, So as to retain the nutrition supply of plant to greatest extent so as to reduce influence to seed production or without influence.

In order to solve the above technical problems, present invention firstly provides a kind of reagent set, be denoted as reagent set 1, it is described at Set reagent 1 is made of the promoter of entitled STK and the protein of entitled bin2-1；

The STK be following a1) a2) or a3):

A1) DNA molecular shown in 1-3093 of sequence 1 in sequence table；

A2 the nucleotide sequence) and a1) limited has 75% or 75% or more identity, and DNA with the same function Molecule；

A3) the nucleotide sequence hybridization limited under strict conditions with a1) or a2), and DNA with the same function points Son；

The bin2-1 is following A1), A2) or A3):

A1) amino acid sequence is the protein of sequence 2；

A2) by amino acid sequence shown in sequence 2 in sequence table by one or several amino acid residues substitution and/or Deletion and/or addition and protein with the same function；

A3) in A1) or the obtained fused protein of N-terminal A2) or/and C-terminal connection label.

Term " identity " used herein refers to the sequence similarity with native sequence nucleic acid." identity " includes and this hair Bright nucleotide sequence has the core of 75% or higher or 85% or higher or 90% or higher or 95% or higher identity Nucleotide sequence.Identity can with the naked eye or computer software is evaluated.Using computer software, two or more sequences it Between identity can be indicated with percentage (%), can be used to evaluate identity between correlated series.

The stringent condition can be as follows: 50 DEG C, in 7% lauryl sodium sulfate (SDS), 0.5M NaPO₄And 1mM Hybridize in the mixed solution of EDTA, is rinsed in 50 DEG C, 2 × SSC, 0.1%SDS；May be used also are as follows: 50 DEG C, in 7%SDS, 0.5M NaPO₄Hybridize in the mixed solution of 1mM EDTA, is rinsed in 50 DEG C, 1 × SSC, 0.1%SDS；May be used also are as follows: 50 DEG C, 7%SDS, 0.5M NaPO₄Hybridize in the mixed solution of 1mM EDTA, is rinsed in 50 DEG C, 0.5 × SSC, 0.1%SDS；Also It can are as follows: 50 DEG C, in 7%SDS, 0.5M NaPO₄Hybridize in the mixed solution of 1mM EDTA, at 50 DEG C, 0.1 × SSC, 0.1% It is rinsed in SDS；May be used also are as follows: 50 DEG C, in 7%SDS, 0.5M NaPO₄Hybridize in the mixed solution of 1mM EDTA, at 65 DEG C, It is rinsed in 0.1 × SSC, 0.1%SDS；It can also are as follows: in 6 × SSC, the solution of 0.5%SDS, hybridize at 65 DEG C, then with 2 × SSC, 0.1%SDS and 1 × SSC, it is primary that 0.1%SDS respectively washes film；It can also are as follows: in the solution of 2 × SSC, 0.1%SDS, 68 Hybridize at DEG C and wash film 2 times, each 5min, and in 0.5 × SSC, the solution of 0.1%SDS, hybridizes at 68 DEG C and wash film 2 It is secondary, each 15min；Can also are as follows: 0.1 × SSPE (or 0.1 × SSC), 0.1%SDS solution in, hybridize and wash under the conditions of 65 DEG C Film.

Above-mentioned 75% or 75% or more identity can be 80%, 85%, 90% or 95% or more identity.

In order to make A1) in protein convenient for purifying, amino acid sequence shown in sequence 2 can be formed in by sequence table The upper label as shown in the table of amino terminal or carboxyl terminal connection of protein.

Table: the sequence of label

Label	Residue	Sequence
			Poly-Arg	5-6 (usually 5)	RRRRR
Poly-His	2-10 (usually 6)	HHHHHH
			FLAG	8	DYKDDDDK
Strep-tag II	8	WSHPQFEK
			c-myc	10	EQKLISEEDL

Above-mentioned A2) in bin2-1 protein, to have 75% or 75% with the amino acid sequence of protein shown in sequence 2 The above identity and protein with the same function.It is described that there is 75% or 75% or more identity to be with 75%, have 80%, with 85%, with 90%, with 95%, with 96%, with 97%, with 98% or with 99% identity.

Above-mentioned A2) in bin2-1 protein can be artificial synthesized, can also first synthesize its encoding gene, then carry out biological expression It obtains.

Above-mentioned A2) in the encoding gene of bin2-1 protein can be by will be in DNA molecular shown in sequence 3 in sequence table The codon of one or several amino acid residues is lacked, and/or carries out the missense mutation of one or several base-pairs, and/or The coded sequence that its 5 ' end and/or 3 ' ends connect label shown in table obtains.Wherein, DNA shown in sequence 3 points in sequence table Bin2-1 protein shown in sub- coded sequence 2.

A3) fused protein can be the fused protein of bin2-1 protein and GFP.

The present invention also provides another reagent set, be denoted as reagent set 2, the reagent set 2 by the STK and with Relevant biomaterial (the being denoted as biomaterial first) composition of the bin2-1；

The biomaterial is any one of following B1) to B7):

B1 the nucleic acid molecules of the bin2-1) are encoded；

B2) contain B1) expression cassettes of the nucleic acid molecules；

B3) contain B1) recombinant vectors of the nucleic acid molecules or contain B2) recombinant vector of the expression cassette；

B4) contain B1) recombinant microorganisms of the nucleic acid molecules or contain B2) recombinant microorganism of the expression cassette or Contain B3) recombinant microorganism of the recombinant vector；

B5) contain B1) the transgenic plant cells systems of the nucleic acid molecules or contain B2) transgenosis of the expression cassette Plant cell；

B6) contain B1) Transgenic plant tissues of the nucleic acid molecules or contain B2) transgenosis of the expression cassette plants Object tissue；

B7) contain B1) the genetically modified plants organs of the nucleic acid molecules or contain B2) transgenosis of the expression cassette plants Sundries official.

In above-mentioned reagent set 2, B1) nucleic acid molecules can be following b11)-b15) any one of:

B11) coded sequence is the cDNA molecule or DNA molecular of sequence 3 in sequence table；

B12) DNA molecular shown in sequence 3 in sequence table；

B13) DNA molecular shown in 3100-5536 of sequence 1 in sequence table；

B14) and b11) or b12) or the nucleotide sequence that b13) limits there is 75% or 75% or more identity, and encode The cDNA molecule or DNA molecular of the bin2-1；

B15) under strict conditions with b11) or b12) or b13) or the nucleotide sequence hybridization that b14) limits, and encode institute State the cDNA molecule or DNA molecular of bin2-1；

Any one of B2) expression cassette is following b21)-b23):

B21) DNA molecular shown in sequence 1 in sequence table；

B22 the nucleotide sequence) and b21) limited has 75% or 75% or more identity, and with the same function DNA molecular；

B23) the nucleotide sequence hybridization limited under strict conditions with b21) or b22), and DNA with the same function Molecule.

In above-mentioned application, B2) described in the nucleic acid molecules containing coding bin2-1 protein expression cassette (bin2-1 gene Expression cassette), it is the DNA for referring to express bin2-1 protein in host cell, which not only may include starting bin2-1 base Because of the promoter of transcription, it may also include the terminator for terminating bin2-1 genetic transcription.Further, the expression cassette may also include increasing Hadron sequence.Promoter for use in the present invention can be the STK.

The recombinant vector of the bin2-1 expression casette can be contained with existing expression vector establishment.The plant expression Carrier includes double base agrobacterium vector and the carrier etc. that can be used for plant micropellet bombardment.As pAHC25, pBin438, pCAMBIA1302、pCAMBIA2301、pCAMBIA1301、pCAMBIA1300、pBI121、pCAMBIA1391-Xa、PSN1301 Or pCAMBIA1391-Xb (CAMBIA company) etc..The plant expression vector also may include 3 ' end non-translational regions of foreign gene Domain, i.e., comprising polyadenylation signals and any other DNA fragmentation for participating in mRNA processing or gene expression.The polyadenylic acid letter Number bootable polyadenylic acid is added to 3 ' ends of mRNA precursor, as Agrobacterium crown gall nodule induces (Ti) plasmid gene (such as nopaline Synthase gene Nos), plant gene (such as soybean storage protein genes) 3 ' end transcription non-translational region all have similar functions. When using gene constructed plant expression vector of the invention, enhancer, including translational enhancer or transcriptional enhancer also can be used, These enhancer regions can be ATG initiation codon or neighboring region initiation codon etc., but must read with coded sequence Frame is identical, to guarantee the correct translation of entire sequence.The source of the translation control signal and initiation codon be it is extensive, Can be it is natural, be also possible to synthesis.Translation initiation region can come from transcription initiation region or structural gene.In order to just In transgenic plant cells or plant are identified and screened, plant expression vector used can be processed, it can as being added The coding expressed in plant can produce the enzyme of color change or gene (gus gene, luciferase genes of luminophor Deng), the marker gene of antibiotic (if assigned the nptII gene to kanamycins and associated antibiotic resistance, assigns to herbicide The bar gene of phosphinothricin resistance assigns the hph gene to antibiotic hygromycin resistance, and assigns to methotrexate resistance Dhfr gene is assigned to the EPSPS gene of glyphosate) or (such as anti-herbicide base such as anti-chemical reagent marker gene Cause), provide metabolism mannose ability mannose-6-phosphate isomerase gene.It, can not from the security consideration of genetically modified plants Add any selected marker, transformed plant is directly screened with adverse circumstance.

In above-mentioned application, the carrier can be plasmid, sticking grain, bacteriophage or viral vectors.The plasmid is concretely pCAMBIA1302。

B3) the recombinant vector concretely pSTK-bin2-1-GFP.The pSTK-bin2-1-GFP is will DNA fragmentation between EcoRI and NcoI the identification sequence of pCAMBIA1302 replaces with 1-3093 institutes of sequence 1 in sequence table The STK promoter shown, and utilize the 3100- of XbaI sequence 1 in insetion sequence table in the multiple cloning sites of pCAMBIA1302 The recombinant vector that the encoding gene of bin2-1 protein obtains shown in 5536.PSTK-bin2-1-the GFP contains sequence 1 Shown in expression cassette.

In above-mentioned application, the microorganism can be yeast, bacterium, algae or fungi.Wherein, bacterium can be Agrobacterium, such as agriculture Bacillus GV3101.

In above-mentioned application, the transgenic plant cells system, Transgenic plant tissue and genetically modified plants organ are not wrapped Include propagation material.

The reagent set 1 and the reagent set 2 all have following any purposes:

D1) regulate and control plant plant height；

D2) preparation regulation plant strain high product；

D3 plant plant height) is reduced；

D4) preparation reduces plant strain high product.

The plant can be dicotyledon or monocotyledon.

The present invention also provides a kind of DNA molecular (i.e. bin2-1 expression casette) or with the bin2-1 expression casette Relevant biomaterial, the bin2-1 expression casette be following c1) c2) or c3):

C1) DNA molecular shown in sequence 1 in sequence table；

C2 the nucleotide sequence) and c1) limited has 75% or 75% or more identity, and DNA with the same function Molecule；

C3) the nucleotide sequence hybridization limited under strict conditions with c1) or c2), and DNA with the same function points Son；

The biomaterial relevant to the bin2-1 expression casette, is denoted as biomaterial second, is following C1) extremely Any one of C5):

C1) contain the recombinant vector of the bin2-1 expression casette；

C2) recombinant microorganism containing the bin2-1 expression casette or contain C1) recombination of the recombinant vector is micro- Biology；

C3) transgenic plant cells system containing the bin2-1 expression casette or contain C1) recombinant vector Transgenic plant cells system；

C4) Transgenic plant tissue containing the bin2-1 expression casette or contain C1) recombinant vector turns Gene plant tissue；

C5) the genetically modified plants organ containing the bin2-1 expression casette or contain C1) recombinant vector turns Gene plant organ.

C1) expression cassette can be the pSTK-bin2-1-GFP.

The present invention also provides following X1) or method X2):

X1 the method for plant plant height) is reduced, comprising: the encoding gene of the STK and the bin2-1 are imported into receptor and planted In object, STK is made to drive the expression of the encoding gene of the bin2-1, obtains purpose plant, it is described compared with the recipient plant The plant height of purpose plant reduces；

X2) cultivate plant height reduce plant method, comprising: by the encoding gene of the STK and the bin2-1 import by In body plant, STK is made to drive the expression of the encoding gene of the bin2-1, obtains what the plant height compared with the recipient plant reduced Purpose plant.

In the above method, the encoding gene of the STK and the bin2-1 are imported in recipient plant by by sequence table DNA molecular shown in middle sequence 1 is imported in the recipient plant and is realized.

Further, the encoding gene of the STK and the bin2-1 are imported in recipient plant and is carried using the C1) recombination Body converts the recipient plant and realizes.

In the above method, wherein the encoding gene of the bin2-1 can be modified first as follows, then import in recipient plant, To reach better expression effect:

1) it modifies and optimizes according to actual needs, so that gene efficient expression；For example, can be according to recipient plant institute partially The codon of love changes its codon while keeping the amino acid sequence of encoding gene of bin2-1 of the present invention to accord with Close plant-preference；In optimization process, it is desirable that certain G/C content is kept in the coded sequence after optimization, with best real The high level expression of quiding gene in existing plant, wherein G/C content can be 35%, be more than 45%, more than 50% or more than about 60%；

2) gene order of neighbouring initial methionine is modified, so that translation effectively starting；For example, using in plant The effective sequence known is modified；

3) it is connect with the promoter of various plants expression, in favor of its expression in plant；The promoter may include Composing type, induction type, timing adjusting, growth adjustment, Chemical Regulation, tissue are preferably and tissue-specific promoter；Promoter Selection will need with expression time and space and be changed, and also depend on target kind；Such as the specificity of tissue or organ Promoter is expressed, receptor as needed is depending on what period of development；Although demonstrating many from dicotyledon Promoter can act in monocotyledon, and vice versa, but it is desirable to select dicot promoters are used for Expression in dicotyledon, monocotyledonous promoter is for the expression in monocotyledon；

4) it is connect with suitable transcription terminator, can also be improved the expression efficiency of gene of the present invention；Such as from The tml of CaMV, from the E9 of rbcS；Any known available terminator to work in plant can be with the present invention Gene is attached；

5) enhancer sequence is introduced, such as intron sequences (such as from Adhl and bronzel) and viral leader sequence (such as from TMV, MCMV and AMV).

The encoding gene of the bin2-1 is imported using the recombinant expression carrier of the encoding gene containing the bin2-1 Recipient plant.The recombinant expression carrier concretely pSTK-bin2-1-GFP.

The recombinant expression carrier can be by using Ti-plasmids, plant virus carrying agent, directly delivered DNA, microinjection, and electricity is worn The standard biologics technical method such as hole imports plant cell (Weissbach, 1998, Method for Plant Molecular Biology VIII,Academy Press,New York,pp.411-463；Geiserson and Corey,1998,Plant Molecular Biology(2nd Edition).)。

The purpose plant is interpreted as the not only first generation plant comprising bin2-1 albumen or its encoding gene, also includes it Filial generation.For the purpose plant, the gene can be bred in the species, it is also possible to which traditional breeding techniques is by the gene transfer Into other kinds of same species, particularly including in commercial variety.The purpose plant includes seed, callus, complete Plant and cell.

The recipient plant can be dicotyledon or monocotyledon.

The present invention also provides following any applications:

Y1) reagent set 1 or the reagent set 2 are in the application for regulating and controlling plant strain senior middle school；

Y2) the application of the reagent set 1 or the reagent set 2 in preparation regulation plant strain high product；

Y3) reagent set 1 or the reagent set 2 are in the application for reducing plant strain senior middle school；

Y4) the application of the reagent set 1 or the reagent set 2 in preparation reduction plant strain high product；

Y5) the application of the reagent set 1 or the reagent set 2 in plant breeding；

Y6) application of the STK in regulation plant strain senior middle school；

Y7) application of the STK in preparation regulation plant strain high product；

Y8) STK is in the application for reducing plant strain senior middle school；

Y9) STK reduces the application in plant strain high product in preparation；

Y10) application of the STK in plant breeding；

Y11) the bin2-1 expression casette or the biomaterial second are in the application for regulating and controlling plant strain senior middle school；

Y12) the bin2-1 expression casette or the biomaterial second answering in preparation regulation plant strain high product With；

Y13) the bin2-1 expression casette or the biomaterial second are in the application for reducing plant strain senior middle school；

Y14) the bin2-1 expression casette or the biomaterial second answering in preparation reduction plant strain high product With；

Y15) the application of the bin2-1 expression casette or the biomaterial second in plant breeding；

Y16) application of the method in plant breeding.

The plant can be dicotyledon or monocotyledon.

The dicotyledon can be crucifer, such as arabidopsis.

It is demonstrated experimentally that can not influence leaf area while reducing plant plant height using reagent set of the invention, i.e., While not changing leaf blade size to reach the supply for guaranteeing plant nutrition, the plant height for reducing plant prevents plant from lodging.

Detailed description of the invention

Fig. 1 is the phenotype of positive transgenic plant.A is the size and character of lotus throne leaf, bar=1cm；B is whole strain plant Phenotype, bar=2cm.

Specific embodiment

The present invention is further described in detail With reference to embodiment, and the embodiment provided is only for explaining The bright present invention, the range being not intended to be limiting of the invention.Experimental method in following embodiments is unless otherwise specified Conventional method.Material as used in the following examples, reagent, instrument etc., are commercially available unless otherwise specified. Quantitative test in following embodiment, is respectively provided with three repeated experiments, and results are averaged.In following embodiments, such as without special Illustrate, the 1st of each nucleotide sequence is the 5 ' terminal nucleotides of corresponding DNA/RNA in sequence table, and last bit is corresponding 3 ' the terminal nucleotides of DNA/RNA.

PCAMBIA1302 (Zhang Y, Zhang YJ, Yang BJ, Yu XX, Wang D, Zu in following embodiments SH,Xue HW,Lin WH.(2016)Functional characterization of GmBZL2(AtBZR1like gene) reveals the conserved BR signaling regulation in Glycine max.Sci Rep.Scientific Reports 6:31134.) public can obtain the biomaterial from applicant, and which is only It repeats used in related experiment of the invention, not can be used as other purposes and use.

Embodiment 1, encoding gene containing STK promoter and bin2-1 protein expression cassette can not influence blade In the case where reduce plant plant height

It present embodiments provides one and can reduce plant plant height but not influence the expression cassette of blade area and (be denoted as STK- Bin2-1), which contains the encoding gene of the promoter of entitled STK and the protein of entitled bin2-1, sequence For sequence 1 in sequence table.1-3093 of sequence 1 are the sequence of STK promoter；3100-5536 of sequence 1 are Sequence (3100-3114,3704-3796,3888-4220,4313- of sequence 1 of the encoding gene of bin2-1 protein 4387,4581-4537,4632-4772,4863-4913,4994-5089,5176-5259,5342-5536 are exon Sequence, 3115-3703,3797-3887,4221-4312,4388-4480,4538-4631,4773-4862,4914- 4993,5090-5175,5260-5314 sequences for introne), the CDS sequence of the encoding gene is sequence 3 in sequence table, Bin2-1 protein shown in coded sequence 2；6299-6301 of sequence 1 are the sequence of terminator codon.The expression cassette The detecting step of function is as follows:

1, the building of recombinant vector

The encoding gene of artificial synthesized STK promoter and bin2-1 protein, by the EcoRI and NcoI of pCAMBIA1302 DNA fragmentation between identification sequence replaces with STK promoter shown in 1-3093 of sequence 1 in sequence table, and utilizes XbaI In pCAMBIA1302 in insetion sequence table bin2-1 protein shown in 3100-5536 of sequence 1 encoding gene, Recombinant vector is obtained, which is denoted as pSTK-bin2-1-GFP.

PSTK-bin2-1-GFP contains expression cassette shown in sequence 1, and can express bin2-1 protein and GFP merges egg The expression of white matter, the fused protein is driven by STK promoter.

2, the building of transgenic plant

The pSTK-bin2-1-GFP that step 1 is obtained is imported in Agrobacterium GV3101, is turned using the gene of mediated by agriculture bacillus Change method arabidopsis thaliana transformation Col (i.e. Colombia's type arabidopsis) carries out resistance screening using hygromycin and obtains transgenic plant. Using pCAMBIA1302 replacement pSTK-bin2-1-GFP as control, unloaded adjoining tree is constructed.

3, the identification of transgenic plant

Identification on DNA level: primer BIN2-3'-F (5 '-TCTTGAGGTTTCTTCAGTCTCT-3 ') and GFP- are utilized The genomic DNA for the transgenic plant that R (5 '-GAACACCATAAGAGAAAGTAGTG-3 ') obtains step 2 carries out PCR expansion Increase, the plant containing 492bpDNA segment is positive transgenic plant in PCR product, and the plant without containing 492bpDNA segment is Negative transgenic plant.The identification sequence of BIN2-3'-F and GFP-R is located at bin2-1 protein and the encoding gene of GFP In, 3 positive transgenic strains (#31, #32 and #34) are identified in the method.

4, the phenotype of transgenic plant

The plant height of the 50th day positive transgenic strain and the area of lotus throne leaf are sowed in measurement, and utilize arabidopsis col and sky Adjoining tree is carried as control, the result is shown in Figure 1.Each strain measures three plant, and each plant is positive transgenic plant.

The results show that the plant height and lotus throne leaf area of unloaded adjoining tree and Col are without significant difference；With Col phase Than the plant height of #31, #32 and #34 are remarkably decreased, but the shape of lotus throne leaf and area are without significant difference.Show expression cassette STK-bin2-1 can not influence leaf area while reducing arabidopsis plant height, i.e., do not changing leaf blade size to reach and guarantee While the supply of plant nutrition, the plant height for reducing plant prevents plant from lodging.

<110>Shanghai Communications University

<120>for reducing the reagent set and method of plant plant height

<160> 3

<170> PatentIn version 3.5

<210> 1

<211> 6301

<212> DNA

<213>artificial sequence

<400> 1

gctctgcaat ttcacctttc tctttaattt tccaccaaag aaatcttaaa ttttaatatt 60

tgcttctcaa tatataaatt atatatcatt tcaaaattta cattatacat atattaatct 120

caatatgtag gtgtatatat aacacattta tctgtcaact tttcatttcg tttttgttgg 180

gtatgttctc actttcttga aagaatgaaa catatgtacg tacatttacg tggaagttat 240

gtatatgtat taaaatgttt aagttaagta aaaggttagg ttgtataaat gataaatcac 300

acatgtctaa attaaaaaaa aaaacactaa taaccaacga gtggtggtac aattatatat 360

tttgagtggt gcataatttt cttttggatt tatgataggg ttcgtctatt ttactattaa 420

ctaacagtct cacatatttt tactattaat tcatcgttgg tcactatcga cgctttaatt 480

tgcaaattta aacttcatgt attatttagt taatgattag gtcatggatt cttccaatta 540

caaaaattat gcctcaaata tgttccttac atatggctga aaatggttgt aaaaaaaagt 600

taacgaatgt tttttttttt ttactttttc cttattttgt ttctttttac cattttggag 660

aggagattaa ttattataat catcaaaaaa ctgaaaataa cgagaaagta ttacaagaca 720

ataatgtggt aaaatgttga ctgtctcaaa atacaaaatg tcgcttaaga aaggagctat 780

atattttcta ctgctcgcaa tcttagaatt aatttttaaa aataatttca aaattttagt 840

aataatatta tatgttatga actaattaaa ccttaatgta ctgccaatca tattaagaat 900

gaaataaata tttcttattg ttctttgtca aaaaaaaaac tatttcttat tgttttatat 960

attttagtga aactatataa atgttttttt tgaagttttc tttgtaaccg ggaaattaaa 1020

aaaaccgtga ttgcaattgc aaaatttgag attataaatt cataatgata agcctaatca 1080

ctgaaacacc atggttttat tatttcactt ttttttacca tgcaattaat ttttttacct 1140

cattatatat ccagtaatga aagttagaac aattaatata attgcatctt cggagatccc 1200

aaatcttggt cttgccgtga acttggccaa agtatgaaat gatggcgcat gtagcttagc 1260

tagagtctcc atttctctac aaaaataaga attaaacaac aaatactcac acttaagagt 1320

tttgtaggaa tcttaaatac tactaatact ttatatgtgc gatttagctt aaaagaaagg 1380

ttaaaataac tttttttggt tgctcgttat tttttccttt cattcttaaa attttttcta 1440

ttcaaaatta agattcaggt ttttttttat aatatagcaa atgcattggc aagtggcaac 1500

cgcaaccgca gcaatcgcaa tctcactttg gacattggaa ggtgagcaaa gaaagaaacg 1560

tctacaaata gacatattga gtgagaagct tagggttttt gaagagaaaa gaatttgggt 1620

ttgtttttgg gattctcaaa aaactcaaaa taaactttcc tcattttttt tttctttctt 1680

ttgttttgtt ttctttataa aggagaaaga aagagaaaga gagtttccga aagctgaatt 1740

gagttgggtg aagcaaattc tcaggtctgt ctgtcatgtc tttacttctt cttctcataa 1800

aggaaaacac ttcttgatct catcaagttc ccatctttgt acacatctct tcattcaaaa 1860

actctacatt tatatctcta tatatgcaga ttccaagctt gtaataattc ttgtcatcat 1920

gggttttttt cttgccttgt ctctccagaa aacttaattt tcacaattat gaattatttt 1980

ccttattttt gttactttct tctgtttgat gctctttccc catgaaagaa agaaagagaa 2040

agaaagatca caagggtttt atttggtcac cagattaaga attaggttat cttttttttt 2100

tttttagttt ctcacatgta gatctctgct agattctctt tcctttcttg tcagctaaag 2160

tggagttttt gagtctgaaa atcttccaaa tctgttgtat ttcttcttcc tctcttctcg 2220

aaactaagca tggttttgtc tctctcttct ttctgtgtaa taatgtttct tgtgtttccc 2280

ttatgtacta taacttcagt ttcatgtgca tcagtgcctt cttcttcttc ctcttcaata 2340

tcaatttgat ttgttttctc tctccatatt tccaattttt tttctttatc aaaagttatt 2400

taatcttttg ctctgtgaaa acaaactaaa gaatgcgtaa gaatgcctac atacatcaga 2460

tcaatgaatt tgtaagacat attacatatg tctaaagtat ttacatcttc cagatctaag 2520

atcattacgt atatgttgtt ttccattttg accccgtgaa gccttataga catttattga 2580

agagggaggg agatagtaca cgggatgggt caaaattatt tgtatctggg taccaaaaaa 2640

ccatgtaata tattgtcaga ttttatctcc ctggctttaa aaaaacaatt cctgatgagt 2700

tagttaagta tatagataat acatatagaa cctatctagc taacaagaat tacatgtaat 2760

atatattaat aatcttgtaa tttgaataaa ttcttaaact ctagttccaa caacatcaga 2820

ttcagttgat tcttgtctgt ccttgagatc aatcaacaac atccttaatt gaatttcccg 2880

agaaagttgg atacagaaag aaaacttttt ttttaatata tataacaaaa gtcgagtttg 2940

gtatgtatta attaagtaca ttaattcaaa ccctagcttt agtttttttt ttatcttctt 3000

gcagtctgcc actagtttgt gtgttaatat ttatcttacc aatactgccg aaattgagca 3060

attctaagtg ttgtttgatg tttaaaatga aggtctagaa tggctgatga taaggtaaag 3120

ctgcttgtgt tccttttgct gtcttttgaa gaagaagatc ctgttttttg gtttttccac 3180

atttgaccct tcaattagtt gtcttgagat tcctgttctt acagatgttt tgtgataata 3240

aatctagttt agttagtact cttgaagttg aacggttttg agttctgggt ttgtccaaag 3300

ttttgagctt tcgttaactt tttgacttac cctgagatct ctgagggttt tgagttctga 3360

gcttagaatt ttctaagtta gctctgttgg gatgatccat gtctatatat ctcgatctgt 3420

gattaatcca gagtttatac aagctgctag atccataatt gaacatagat tagtccttgt 3480

ttggtttgtt atatgtattt gttttgttta ccattctttt ggcgtgacaa aagtatatat 3540

ttttagtttt aactaaatca gattcactct gcgtaacggt tattttgtaa ccactctttt 3600

aggataaaag tttccttctt tagacttttg attctcagac aagcattatt cttttagctt 3660

ttgataatgg ttttgtgctg atattaaagc ttttctcttt caggagatgc ctgctgctgt 3720

agttgatgga catgatcaag tcactggtca tattatttcc accacaatcg gtggcaaaaa 3780

tggtgaacca aaacaggtat ttttaaggct tttaaccaaa tagactcact tttatgtata 3840

tgcaagattt tgatggttac caatacattt ttctatgttg ttgttagaca attagttaca 3900

tggcggagcg agttgttggt acaggctcgt tcgggatcgt tttccaagca aaatgtttgg 3960

agactggaga aaccgtggcg ataaagaagg ttttgcaaga tagaagatac aagaaccgag 4020

aacttcagtt gatgcgtgtg atggatcatc cgaatgtggt ttgtttgaag cattgcttct 4080

tttcgactac aagtaaagac gagcttttct tgaacttggt tatggagtat gtccctgaga 4140

gcttgtatcg agttctgaaa cattatagta gtgcaaacca aagaatgcct cttgtctatg 4200

ttaaacttta catgtatcag gtaataacaa cacacattca tatcttccat ttccaaagtt 4260

gatgtacata agattgttct taggctgata ttacttcctt ttgttatgtt agatcttccg 4320

gggacttgct tacattcaca atgttgctgg agtttgtcac agagatctaa agcctcaaaa 4380

tcttctggta tgtgtaacat tttaagattg aactctttgt tttttttctt gcctttgttt 4440

ctttcgttct taatgtatct cttctggtct ctttctatag gttgatcctc ttactcatca 4500

agtcaaaatc tgtgactttg gcagtgcgaa acagctcgta agactttgtg acatataaac 4560

tcattcgact tgtagcggtt gttgttttct gtgatcttgt catttactgt tgaaatttac 4620

ttttgcttca ggttaaaggt gaagccaaca tttcttacat ctgctcacga ttctaccgtg 4680

cacccgagct catatttggt gccactgagt acacaacttc tattgatatc tggtctgctg 4740

gttgtgttct tgctgagctt cttcttggtc aggtaaacaa ttctttcagt aaccagctta 4800

ttcaatctcc atgtgtatat ttgcattagg actcattgtg actatatcat gttcttatgc 4860

agccattatt tcccggagaa aatgctgtgg atcagctcgt tgaaattata aaagtaagaa 4920

tctttaaacg atgattcctt gcaaattaca ttctttggct acaaaatcct cactgtatag 4980

ttgttgtaca caggttcttg gtacaccaac tcgaaaagaa atccgttgta tgaatccaca 5040

ttacacagat ttcaggtttc cacagataaa ggcacatccc tggcacaagg ttagtgtctt 5100

ttctcttttt gcatgtgttc ttgtttcagt ttctttcttc acacatcaac tgatcataat 5160

tacgttttgg tttagatctt ccacaaaagg atgcccccag aagcgattga ttttgcatca 5220

aggctgcttc aatactctcc aagtctaaga tgcacagcgg taagcattgg tcttgaggtt 5280

tcttcagtct ctaagaatcc aactgaatcc ttactatata ttttgtttcc tcgtatttca 5340

gctcgaagct tgtgcacatc cgttctttga tgaactcaga gaaccaaacg ctcgtttacc 5400

aaatggacgg cctttcccgc ctctcttcaa cttcaaacaa gaagtagctg gatcatcacc 5460

tgaactggtc aacaagttga ttccagacca tatcaagaga caattgggtc taagcttctt 5520

gaatcaatct ggaacttcta gagccatggt agatctgact agtaaaggag aagaactttt 5580

cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc acaaattttc 5640

tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta aatttatttg 5700

cactactgga aaactacctg ttccgtggcc aacacttgtc actactttct cttatggtgt 5760

tcaatgcttt tcaagatacc cagatcatat gaagcggcac gacttcttca agagcgccat 5820

gcctgaggga tacgtgcagg agaggaccat cttcttcaag gacgacggga actacaagac 5880

acgtgctgaa gtcaagtttg agggagacac cctcgtcaac aggatcgagc ttaagggaat 5940

cgatttcaag gaggacggaa acatcctcgg ccacaagttg gaatacaact acaactccca 6000

caacgtatac atcatggccg acaagcaaaa gaacggcatc aaagccaact tcaagacccg 6060

ccacaacatc gaagacggcg gcgtgcaact cgctgatcat tatcaacaaa atactccaat 6120

tggcgatggc cctgtccttt taccagacaa ccattacctg tccacacaat ctgccctttc 6180

gaaagatccc aacgaaaaga gagaccacat ggtccttctt gagtttgtaa cagctgctgg 6240

gattacacat ggcatggatg aactatacaa agctagccac caccaccacc accacgtgtg 6300

a 6301

<210> 2

<211> 380

<212> PRT

<213>artificial sequence

<400> 2

Met Ala Asp Asp Lys Glu Met Pro Ala Ala Val Val Asp Gly His Asp

1 5 10 15

Gln Val Thr Gly His Ile Ile Ser Thr Thr Ile Gly Gly Lys Asn Gly

20 25 30

Glu Pro Lys Gln Thr Ile Ser Tyr Met Ala Glu Arg Val Val Gly Thr

35 40 45

Gly Ser Phe Gly Ile Val Phe Gln Ala Lys Cys Leu Glu Thr Gly Glu

50 55 60

Thr Val Ala Ile Lys Lys Val Leu Gln Asp Arg Arg Tyr Lys Asn Arg

65 70 75 80

Glu Leu Gln Leu Met Arg Val Met Asp His Pro Asn Val Val Cys Leu

85 90 95

Lys His Cys Phe Phe Ser Thr Thr Ser Lys Asp Glu Leu Phe Leu Asn

100 105 110

Leu Val Met Glu Tyr Val Pro Glu Ser Leu Tyr Arg Val Leu Lys His

115 120 125

Tyr Ser Ser Ala Asn Gln Arg Met Pro Leu Val Tyr Val Lys Leu Tyr

130 135 140

Met Tyr Gln Ile Phe Arg Gly Leu Ala Tyr Ile His Asn Val Ala Gly

145 150 155 160

Val Cys His Arg Asp Leu Lys Pro Gln Asn Leu Leu Val Asp Pro Leu

165 170 175

Thr His Gln Val Lys Ile Cys Asp Phe Gly Ser Ala Lys Gln Leu Val

180 185 190

Lys Gly Glu Ala Asn Ile Ser Tyr Ile Cys Ser Arg Phe Tyr Arg Ala

195 200 205

Pro Glu Leu Ile Phe Gly Ala Thr Glu Tyr Thr Thr Ser Ile Asp Ile

210 215 220

Trp Ser Ala Gly Cys Val Leu Ala Glu Leu Leu Leu Gly Gln Pro Leu

225 230 235 240

Phe Pro Gly Glu Asn Ala Val Asp Gln Leu Val Glu Ile Ile Lys Val

245 250 255

Leu Gly Thr Pro Thr Arg Lys Glu Ile Arg Cys Met Asn Pro His Tyr

260 265 270

Thr Asp Phe Arg Phe Pro Gln Ile Lys Ala His Pro Trp His Lys Ile

275 280 285

Phe His Lys Arg Met Pro Pro Glu Ala Ile Asp Phe Ala Ser Arg Leu

290 295 300

Leu Gln Tyr Ser Pro Ser Leu Arg Cys Thr Ala Leu Glu Ala Cys Ala

305 310 315 320

His Pro Phe Phe Asp Glu Leu Arg Glu Pro Asn Ala Arg Leu Pro Asn

325 330 335

Gly Arg Pro Phe Pro Pro Leu Phe Asn Phe Lys Gln Glu Val Ala Gly

340 345 350

Ser Ser Pro Glu Leu Val Asn Lys Leu Ile Pro Asp His Ile Lys Arg

355 360 365

Gln Leu Gly Leu Ser Phe Leu Asn Gln Ser Gly Thr

370 375 380

<210> 3

<211> 1140

<212> DNA

<213>artificial sequence

<400> 3

atggctgatg ataaggagat gcctgctgct gtagttgatg gacatgatca agtcactggt 60

catattattt ccaccacaat cggtggcaaa aatggtgaac caaaacagac aattagttac 120

atggcggagc gagttgttgg tacaggctcg ttcgggatcg ttttccaagc aaaatgtttg 180

gagactggag aaaccgtggc gataaagaag gttttgcaag atagaagata caagaaccga 240

gaacttcagt tgatgcgtgt gatggatcat ccgaatgtgg tttgtttgaa gcattgcttc 300

ttttcgacta caagtaaaga cgagcttttc ttgaacttgg ttatggagta tgtccctgag 360

agcttgtatc gagttctgaa acattatagt agtgcaaacc aaagaatgcc tcttgtctat 420

gttaaacttt acatgtatca gatcttccgg ggacttgctt acattcacaa tgttgctgga 480

gtttgtcaca gagatctaaa gcctcaaaat cttctggttg atcctcttac tcatcaagtc 540

aaaatctgtg actttggcag tgcgaaacag ctcgttaaag gtgaagccaa catttcttac 600

atctgctcac gattctaccg tgcacccgag ctcatatttg gtgccactga gtacacaact 660

tctattgata tctggtctgc tggttgtgtt cttgctgagc ttcttcttgg tcagccatta 720

tttcccggag aaaatgctgt ggatcagctc gttgaaatta taaaagttct tggtacacca 780

actcgaaaag aaatccgttg tatgaatcca cattacacag atttcaggtt tccacagata 840

aaggcacatc cctggcacaa gatcttccac aaaaggatgc ccccagaagc gattgatttt 900

gcatcaaggc tgcttcaata ctctccaagt ctaagatgca cagcgctcga agcttgtgca 960

catccgttct ttgatgaact cagagaacca aacgctcgtt taccaaatgg acggcctttc 1020

ccgcctctct tcaacttcaa acaagaagta gctggatcat cacctgaact ggtcaacaag 1080

ttgattccag accatatcaa gagacaattg ggtctaagct tcttgaatca atctggaact 1140

Claims (10)

1. reagent set is made of the promoter of entitled STK and the protein of entitled bin2-1；

The STK be following a1) a2) or a3):

A1) DNA molecular shown in 1-3093 of sequence 1 in sequence table；

A2 the nucleotide sequence) and a1) limited has 75% or 75% or more identity, and DNA molecular with the same function；

A3) the nucleotide sequence hybridization limited under strict conditions with a1) or a2), and DNA molecular with the same function；

The bin2-1 is following A1), A2) or A3):

A1) amino acid sequence is the protein of sequence 2；

A2) amino acid sequence shown in sequence 2 in sequence table is passed through to the substitution and/or missing of one or several amino acid residues And/or addition and protein with the same function；

A3) in A1) or the obtained fused protein of N-terminal A2) or/and C-terminal connection label.

2. reagent set, the STK as described in claim 1 and biomaterial group relevant to bin2-1 described in claim 1 At；

The biomaterial is any one of following B1) to B7):

B1 the nucleic acid molecules of bin2-1 described in claim 1) are encoded；

B2) contain B1) expression cassettes of the nucleic acid molecules；

B3) contain B1) recombinant vectors of the nucleic acid molecules or contain B2) recombinant vector of the expression cassette；

B4) contain B1) recombinant microorganisms of the nucleic acid molecules or contain B2) recombinant microorganism of the expression cassette or contain B3) the recombinant microorganism of the recombinant vector；

B5) contain B1) the transgenic plant cells systems of the nucleic acid molecules or contain B2) genetically modified plants of the expression cassette Cell line；

B6) contain B1) Transgenic plant tissues of the nucleic acid molecules or contain B2) the genetically modified plants group of the expression cassette It knits；

B7) contain B1) the genetically modified plants organs of the nucleic acid molecules or contain B2) the genetically modified plants device of the expression cassette Official.

3. reagent set according to claim 2, it is characterised in that: B1) nucleic acid molecules are following b11)-b15) in It is any:

B11) coded sequence is the cDNA molecule or DNA molecular of sequence 3 in sequence table；

B12) DNA molecular shown in sequence 3 in sequence table；

B13) DNA molecular shown in 3100-5536 of sequence 1 in sequence table；

B14) and b11) or b12) or the nucleotide sequence that b13) limits there is 75% or 75% or more identity, and encode right It is required that the cDNA molecule or DNA molecular of bin2-1 described in 1；

B15) under strict conditions with b11) or b12) or b13) or the nucleotide sequence hybridization that b14) limits, and encode right and want Ask the cDNA molecule or DNA molecular of bin2-1 described in 1；

Any one of B2) expression cassette is following b21)-b23):

B21) DNA molecular shown in sequence 1 in sequence table；

B22 the nucleotide sequence) and b21) limited has 75% or 75% or more identity, and DNA with the same function points Son；

B23) the nucleotide sequence hybridization limited under strict conditions with b21) or b22), and DNA molecular with the same function.

4. reagent set according to claim 1 to 3, it is characterised in that: the reagent set has following any Purposes:

D1) regulate and control plant plant height；

D2) preparation regulation plant strain high product；

D3 plant plant height) is reduced；

D4) preparation reduces plant strain high product.

5. reagent set according to claim 4, it is characterised in that: the plant is that dicotyledon or unifacial leaf are planted Object.

6.DNA molecule or biomaterial relevant to the DNA molecular, the DNA molecular be following c1) c2) or c3):

C1) DNA molecular shown in sequence 1 in sequence table；

C2 the nucleotide sequence) and c1) limited has 75% or 75% or more identity, and DNA molecular with the same function；

C3) the nucleotide sequence hybridization limited under strict conditions with c1) or c2), and DNA molecular with the same function；

The biomaterial relevant to the DNA molecular is following C1) any one of to C5):

C1) contain the recombinant vector of the DNA molecular；

C2) recombinant microorganism containing the DNA molecular or contain C1) recombinant microorganism of the recombinant vector；

C3) the transgenic plant cells system containing the DNA molecular or contain C1) genetically modified plants of the recombinant vector are thin Born of the same parents system；

C4) Transgenic plant tissue containing the DNA molecular or contain C1) Transgenic plant tissue of the recombinant vector；

C5) the genetically modified plants organ containing the DNA molecular or contain C1) the genetically modified plants organ of the recombinant vector.

7. following X1) or method X2):

X1 the method for plant plant height) is reduced, comprising: import the encoding gene of STK described in claim 1 and the bin2-1 In recipient plant, STK is made to drive the expression of the encoding gene of the bin2-1, obtain purpose plant, with the recipient plant phase Than the plant height of the purpose plant reduces；

X2 the method that plant height reduces plant) is cultivated, comprising: by the encoding gene of STK described in claim 1 and the bin2-1 It imports in recipient plant, so that STK is driven the expression of the encoding gene of the bin2-1, obtain the plant height compared with the recipient plant Reduced purpose plant.

8. according to the method described in claim 7, it is characterized by: by the volume of STK described in claim 1 and the bin2-1 It is realized in code channel genes recipient plant by importing DNA molecular shown in sequence 1 in sequence table in the recipient plant.

9. following any applications:

Y1) application of the reagent set as claimed in any one of claims 1-3 in regulation plant strain senior middle school；

Y2) application of the reagent set as claimed in any one of claims 1-3 in preparation regulation plant strain high product；

Y3) reagent set as claimed in any one of claims 1-3 is in the application for reducing plant strain senior middle school；

Y4) reagent set as claimed in any one of claims 1-3 reduces the application in plant strain high product in preparation；

Y5) application of the reagent set as claimed in any one of claims 1-3 in plant breeding；

Y6) application of the STK described in claim 1 in regulation plant strain senior middle school；

Y7) application of the STK described in claim 1 in preparation regulation plant strain high product；

Y8) STK described in claim 1 is in the application for reducing plant strain senior middle school；

Y9) STK described in claim 1 reduces the application in plant strain high product in preparation；

Y10) application of the STK described in claim 1 in plant breeding；

Y11) DNA molecular described in claim 6 or the biomaterial are in the application for regulating and controlling plant strain senior middle school；

Y12) the application of DNA molecular described in claim 6 or the biomaterial in preparation regulation plant strain high product；

Y13) DNA molecular described in claim 6 or the biomaterial are in the application for reducing plant strain senior middle school；

Y14) the application of DNA molecular described in claim 6 or the biomaterial in preparation reduction plant strain high product；

Y15) the application of DNA molecular described in claim 6 or the biomaterial in plant breeding；

Y16) the application of claim 7 or 8 the methods in plant breeding.

10. method according to claim 7 or 8 or application as claimed in claim 9, it is characterised in that: the receptor is planted Object is dicotyledon or monocotyledon；

The plant is dicotyledon or monocotyledon.