CN101560251A - Associated protein for plant root growth and encoding gene and application thereof - Google Patents

Abstract

The invention discloses an associated protein for plant root growth and an encoding gene and application thereof. The protein is provided with one of the following amino acid residue sequences: (1) SEQ ID No.3 amino acid residue sequence in the sequence table; and (2) protein which is formed by substituting and/or deleting and/or adding one or a few SEQ ID No.3 amino acid residues in the sequence table and has associated protein activity for plant root growth. The associated protein for plant root growth and the encoding gene thereof can promote the growth of the plant root, and play great role in promoting the root system of plant, improving the yield and improving the variety of the plant (particularly crops).

Description

Associated protein for plant root growth and encoding gene thereof and application

Technical field

The present invention relates to a kind of associated protein for plant root growth and encoding gene thereof and application, particularly relate to associated protein for plant root growth and encoding gene and their application in growth and development of plants that derives from Arabidopis thaliana.

Background technology

As higher organism, the difference of the maximum of Plants and Animals be plant must set growth, the fixing and nutritive substance of this growth draw the growth that must depend on root system of plant consumingly.So the good growth of growing for plant integral body of root system plays crucial effects.No matter be monocotyledons or dicotyledons, though the structure of root system and form are different, all to form for sophisticated by root cap district, meristematic zone, elongation zone and maturation zone four parts, wherein every part is all being carried out different functions.Root cap is made up of central root cap cell and side root cap cell, and along with the continuous growth of the tip of a root, thereby root cap cell detachment has foremost played the effect of protection to the tip of a root; Central root cap cell contains a large amount of starch small graiies simultaneously, plays the effect of experiencing gravity, thereby makes the root of plant produce geotropic growth.Meristematic zone is made up of the cell that is in division stage, and the continuous division of these cells just makes the number of cell increase, and root is constantly grown.The elongation zone is made up of the cell between splitting status and differentiation state, is the transitory stage between the two.The maturation zone is made up of well differentiated cell, and the cell size and form of these differentiation and maturations no longer changes, and carries out different functions separately.

The stem cell that is called as initiating cell in the Arabidopis thaliana root meristematic tissue has produced various types of cells in every confluent monolayer cells layer by the spatial division, and these cells have been formed a vegetative clone.One in two daughter cells that initiating cell produces through division is once still keeping primitiveness, and the another one cell becomes transit-amplifyingcell (TA cell), this cell through division many times after, be differentiated to form self characteristics.The cell that is centered around the quiescent center (QC) of initiating cell central authorities seldom divides and stem cell has on every side been formed the merismatic stem cell of root habitat (stem cell niche), and QC is the organization center of this niche.QC by and the signal communication of stem cell on every side, suppress its differentiation, the primitiveness that keeps stem cell, at present very few for the understanding of these signaling molecule essence, but from root meristematic tissue and the merismatic contrast of bud, find, the initiating cell of root is similar to the merismatic stem cell of bud, and QC is similar to the OC cell of expressing WUS.

In the research of some mutant of regulating and control the Arabidopis thaliana root growth and gene thereof, find that the similar signal transduction path of CLV has also participated in the regulation and control of root stem cell, and the isolation identification of some other mutant, as wherein shortroot (shr) and scarecrow (scr) mutant all show as the root weak point, and these two site mutation have all caused root to lack a confluent monolayer cells between epidermis and endodermis, and it is necessary that SHR and SCR express for the characteristic of keeping QC simultaneously.In addition, the PLT gene also is necessary for the establishment in root stem cell habitat.And PLT and SHR/SCR are two parallel approach in the formation that participates in regulation and control root meristematic tissue stem cell, have regulated and control merismatic formation and growth from embryo's radicle in period to the matured root in seedling stage jointly.

Summary of the invention

The purpose of this invention is to provide a kind of associated protein for plant root growth and encoding gene thereof and application.

Associated protein for plant root growth provided by the present invention, name is called ARD1, derives from Arabidopis thaliana (Arabidopsis thaliana), is following (a) or protein (b):

(a) protein of forming by the amino acid residue sequence of sequence in the sequence table 3;

(b) with the amino acid residue sequence of sequence in the sequence table 3 through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and have the root growth adjusting function by (a) deutero-protein.

Wherein, the sequence in the sequence table 3 is made up of 500 amino-acid residues.

The encoding gene of above-mentioned associated protein for plant root growth (ARD1) also belongs to protection scope of the present invention.

The cDNA gene of above-mentioned associated protein for plant root growth can have one of following nucleotide sequence:

1) dna sequence dna of sequence 1 in the sequence table;

2) polynucleotide of protein sequence shown in the sequence 3 in the code sequence tabulation;

3) nucleotide sequence of the dna sequence dna hybridization that under the rigorous condition of height, can limit with sequence in the sequence table 1.

Wherein, sequence 1 is made up of 1925 deoxynucleotides in the sequence table, and 5 of sequence 1 ' end 268-1770 position nucleotides sequence is classified encoding sequence (ORF) as in sequence table, and coding has the protein of the amino acid residue sequence of sequence 3 in the sequence table.

The genomic gene of above-mentioned associated protein for plant root growth can have one of following nucleotide sequence:

1) dna sequence dna of sequence 2 in the sequence table;

2) polynucleotide of protein sequence shown in the sequence 3 in the code sequence tabulation;

3) nucleotide sequence of the dna sequence dna hybridization that under the rigorous condition of height, can limit with sequence in the sequence table 2.

Wherein, sequence 2 is made up of 4881 Nucleotide in the sequence table, Nucleotide is first exon of this genomic gene from 5 ' end 262-506 position, Nucleotide is first intron of this genomic gene from 5 ' end 507-748 position, Nucleotide is second exon of this genomic gene from 5 ' end 749-820 position, Nucleotide is second intron of this genomic gene from 5 ' end 821-916 position, Nucleotide is the 3rd exon of this genomic gene from 5 ' end 917-1093 position, Nucleotide is the 3rd intron of this genomic gene from 5 ' end 1094-1178 position, Nucleotide is the 4th exon of this genomic gene from 5 ' end 1179-1248 position, Nucleotide is the 4th intron of this genomic gene from 5 ' end 1249-1541 position, Nucleotide is the 5th exon of this genomic gene from 5 ' end 1542-1829 position, Nucleotide is the 5th intron of this genomic gene from 5 ' end 1830-1937 position, from 5 ' end 1938-2078 position Nucleotide is the 6th each and every one exon of this genomic gene, Nucleotide is the 6th intron of this genomic gene from 5 ' end 2079-2157 position, Nucleotide is the 7th exon of this genomic gene from 5 ' end 2158-2260 position, Nucleotide is the 7th intron of this genomic gene from 5 ' end 2261-2352 position, Nucleotide is the 8th exon of this genomic gene from 5 ' end 2353-2499 position, Nucleotide is the 8th intron of this genomic gene from 5 ' end 2500-3085 position, Nucleotide is the 9th exon of this genomic gene from 5 ' end 3086-3132 position, Nucleotide is the 9th intron of this genomic gene from 5 ' end 3133-3219 position, Nucleotide is the tenth exon of this genomic gene from 5 ' end 3220-3273 position, Nucleotide is the tenth intron of this genomic gene from 5 ' end 3274-3345 position, Nucleotide is the 11 exon of this genomic gene from 5 ' end 3346-3464 position, Nucleotide is the 11 intron of this genomic gene from 5 ' end 3465-3552 position, Nucleotide is the 12 exon of this genomic gene from 5 ' end 3553-3688 position, Nucleotide is the 12 intron of this genomic gene from 5 ' end 3689-3766 position, and Nucleotide is the 13 exon of this genomic gene from 5 ' end 3767-4092 position.

The rigorous condition of above-mentioned height can be 0.1 * SSPE (or 0.1 * SSC), in the solution of 0.1%SDS, under 65 ℃, hybridize and wash film.

The expression vector, transgenic cell line and the host bacterium that contain the encoding gene of above-mentioned associated protein for plant root growth all belong to protection scope of the present invention.

Utilize plant expression vector, encoding gene (ARD1) importing vegetable cell or tissue with associated protein for plant root growth of the present invention can obtain root development enhanced plant.

When using ARD1 to make up plant expression vector, before its transcription initiation Nucleotide, can add any enhancement type promotor or inducible promoter.For the ease of transgenic plant cells or plant being identified and screening, can process used plant expression vector, as adding selected marker's (gus gene, luciferase genes etc.) that can in plant, express or antibiotic marker thing (gentamicin marker, kantlex marker etc.) with resistance.From the security consideration of transgenic plant, can not add any selected marker, directly with adverse circumstance screening transformed plant.

Carry ARD1 of the present invention plant expression vector can Ti-plasmids, Ri plasmid, plant viral vector, directly DNA conversion, microinjection, electricity be led, conventional biological method transformed plant cells or tissue such as agriculture bacillus mediated by using, and the plant transformed tissue cultivating is become plant.By the plant transformed host both can be monocotyledonss such as paddy rice, corn, wheat, also can be dicotyledonss such as tomato, Arabidopis thaliana, tobacco, cotton.

Associated protein for plant root growth of the present invention and encoding gene thereof can promote growing of roots of plants, to promoting plant root growth, improve plant biomass, play a great role in the improvement of plant (particularly farm crop) kind.

Below in conjunction with specific embodiment the present invention is described in further details.

Description of drawings

Fig. 1 is the growing state of normal wild type plant and ard1 mutant root

Fig. 2 is the map based cloning synoptic diagram of ARD1 gene

Fig. 3 is the upgrowth situation photo of the ard1 mutant of commentaries on classics ARD1 gene

Embodiment

Method therefor is ordinary method if no special instructions among the following embodiment, and the primer synthesizes and examining order is finished by the living worker's biotechnology in Shanghai company limited.

Embodiment 1, of the present inventionly derive from Arabidopis thaliana and acquisition and functional verification thereof root development genes involved ARD1

One, the acquisition of Arabidopis thaliana ARD1 mutant

Screening by to an Arabidopis thaliana (Col-0) T-DNA mutant library obtains a mutant in the root growth defective, called after ARD1, the phenotype of this mutant is mainly: root growth is slow, and root system is undeveloped, and main root shortens, main root only is about 1/3 of a wild-type, as shown in Figure 1.WT is wild-type Arabidopis thaliana (the Col-0 ecotype) among Fig. 1; ARD1 is the ARD1 mutant that above-mentioned screening obtains.

Two, with the map based cloning method obtain from mutant ARD1 with root development genes involved ARD1

Adopt the map based cloning method to separate from the ARD1 mutant of above-mentioned acquisition and root development genes involved ARD1, concrete grammar may further comprise the steps:

1) F that is obtained with ARD1 mutant (Col-0 background) and Ler (the another one ecotype of Arabidopis thaliana is available from ArabidopsisBiological Resource Center) hybridization ₂For segregating population is material, therefrom having identified 2100 has with above-mentioned ARD1 mutation type surface individuals with same and carries out genetic analysis, the ARD1 assignment of genes gene mapping is cloned on (T6D22) to No. 1 chromosomal BAC of Arabidopis thaliana, be about 550Kb with the physical distance of flag F 21M12, as shown in Figure 2.Among Fig. 2, shown in the arrow near adjacent BAC clone.

2) scope of utilizing the method for map based cloning further to dwindle goal gene, last, according to the arabidopsis gene group sequence of announcing, utilize the candidate gene of PCR method amplifying target genes scope, through sequence alignment.The final gene of determining the place, mutational site.This gene has the nucleotide sequence of sequence 2 in the sequence table, with its called after ARD1.Sequence 2 is made up of 4881 Nucleotide in the sequence table, Nucleotide is first exon of this genomic gene from 5 ' end 262-506 position, Nucleotide is first intron of this genomic gene from 5 ' end 507-748 position, Nucleotide is second exon of this genomic gene from 5 ' end 749-820 position, Nucleotide is second intron of this genomic gene from 5 ' end 821-916 position, Nucleotide is the 3rd exon of this genomic gene from 5 ' end 917-1093 position, Nucleotide is the 3rd intron of this genomic gene from 5 ' end 1094-1178 position, Nucleotide is the 4th exon of this genomic gene from 5 ' end 1179-1248 position, Nucleotide is the 4th intron of this genomic gene from 5 ' end 1249-1541 position, Nucleotide is the 5th exon of this genomic gene from 5 ' end 1542-1829 position, Nucleotide is the 5th intron of this genomic gene from 5 ' end 1830-1937 position, from 5 ' end 1938-2078 position Nucleotide is the 6th each and every one exon of this genomic gene, Nucleotide is the 6th intron of this genomic gene from 5 ' end 2079-2157 position, Nucleotide is the 7th exon of this genomic gene from 5 ' end 2158-2260 position, Nucleotide is the 7th intron of this genomic gene from 5 ' end 2261-2352 position, Nucleotide is the 8th exon of this genomic gene from 5 ' end 2353-2499 position, Nucleotide is the 8th intron of this genomic gene from 5 ' end 2500-3085 position, Nucleotide is the 9th exon of this genomic gene from 5 ' end 3086-3132 position, Nucleotide is the 9th intron of this genomic gene from 5 ' end 3133-3219 position, Nucleotide is the tenth exon of this genomic gene from 5 ' end 3220-3273 position, Nucleotide is the tenth intron of this genomic gene from 5 ' end 3274-3345 position, Nucleotide is the 11 exon of this genomic gene from 5 ' end 3346-3464 position, Nucleotide is the 11 intron of this genomic gene from 5 ' end 3465-3552 position, Nucleotide is the 12 exon of this genomic gene from 5 ' end 3553-3688 position, Nucleotide is the 12 intron of this genomic gene from 5 ' end 3689-3766 position, and Nucleotide is the 13 exon of this genomic gene from 5 ' end 3767-4092 position.Its cDNA sequence is the nucleotide sequence of sequence 1 in the sequence table, and 5 of sequence 1 ' end 268-1770 position nucleotides sequence is classified encoding sequence (ORF) as in sequence table, and coding has the protein (ARD1) of the amino acid residue sequence of sequence 3 in the sequence table.

Three, of the present inventionly derive from Arabidopis thaliana and transgenosis functional verification root development genes involved ARD1

Genome sequence and the carrier pCAMBIA1300 of the ARD1 that obtains according to step 1, the ARD1 gene is connected into the recombinant vectors that obtains between the Sma I of carrier pCAMBIA1300 multiple clone site and the Sal I restriction enzyme site, the primer of design amplification ARD1 sequence, primer sequence is as follows:

Primer 1 (upstream primer): 5 '- CCCGGGGTGAAGAACAAAATATTACA-3 ' (line part Nucleotide is the SmaI recognition site);

Primer 2 (downstream primer): 5 '- GGTACCATGGCAAAATAACCTTGAGG-3 ' (line part Nucleotide is Kpn I recognition site).

The genomic dna that extracts Arabidopis thaliana kind Col-0 is as template, under the guiding of primer 1 and primer 2, carry out pcr amplification, reaction finishes the back PCR product is carried out the detection of 1% agarose gel electrophoresis, reclaim the also dna fragmentation of purifying 4881bp, be the ARD1 genomic fragment, show that through order-checking this fragment has the nucleotide sequence of sequence 2 in the sequence table.

With the ARD1 gene fragment of above-mentioned acquisition with restriction endonuclease sma I be connected with the carrier pCAMBIA1300 that cuts through the same enzyme enzyme after Kpn I carries out double digestion, to connect product and check order, will show correct recombinant vectors called after pCAMBIA1300-ARD1 through order-checking.

PCAMBIA1300-ARD1 is transformed ARD1 mutant SALK_009847 (available from Arabidopsis Biological Resource Center under the mediation of agrobacterium tumefaciens, ABRC), use selected marker's Totomycin (containing 50mg/L Totomycin MS substratum) to carry out resistance screening then, screening obtains the commentaries on classics pCAMBIA1300-ARD1 plant that can grow on the substratum that contains the 50mg/L Totomycin.The commentaries on classics pCAMBIA1300-ARD1 plant that obtains with above-mentioned primer 1 and the screening of primer 2 antagonism carries out the PCR Molecular Identification, obtains the PCR evaluation and shows the correct commentaries on classics pCAMBIA1300-ARD1 plant that changes the ARD1 gene fragment over to.

Evaluation shows the correct commentaries on classics pCAMBIA1300-ARD1 plant that changes the ARD1 gene fragment over to PCR, advance phenotype analytical, be about to ARD1 mutant SALK_009847 (available from Arabidopsis Biological ResourceCenter) (ARD1 among Fig. 3) and PCR evaluation and show that the correct commentaries on classics pCAMBIA1300-ARD1 plant (pCAMBIA1300-ARD1 among Fig. 3) that changes the ARD1 gene fragment over to sows simultaneously on the MS substratum, 4 ℃ of dark culturing are after 3 days, transfer to 22 degrees centigrade illumination cultivation 8 days, as shown in Figure 3, the result shows, it is better than the upgrowth situation of not genetically modified ARD1 mutant to change the pCAMBIA1300-ARD1 plant, shows that the sudden change of ARD1 gene influences growing of root system of plant really.

Sequence table

<160>3

<210>1

<211>1925

<212>DNA

＜213〉Arabidopis thaliana (Arabidopsis thaliana)

<400>1

ctcagtgcgt?gtttataatt?tttctgggtg?gcgagaaacc?aagagaaaat?ctcgtgaccc 60

tgttctctcg?ggagcgcaca?caatctctct?tcttcttcgt?cttcctcctc?caatcgatgt 120

aatcacacac?acacacaaaa?acacatcttt?tccttgtttt?cgtaggacct?agtagatagc 180

gagttttgcc?cccaaaacag?cgatcgaggg?aatttatgag?cacgagatct?cgattcccca 240

tacactaggg?tttggaatcg?aggcacaatg?caaatgaact?ctgtttggaa?gctgtctctt 300

gggttattac?ttcttagctc?agttattggc?tcttttgcgg?aacttgattt?tggccattgc 360

gaaactcttg?tgaaaaaatg?ggctgattct?tcttcatctc?gtgaagaaca?tgttaataaa 420

gacaaacgct?cgcttaagga?tttgctcttc?tttctccacg?ttccgcgaac?tggaggcaga 480

acatattttc?attgtttttt?gaggaagttg?tatgatagct?ctgaggaatg?tcctcgatct 540

tacgacaagc?tccacttcaa?tccaaggaag?gaaaagtgca?agttgttagc?cacacatgat 600

gattatagtt?tgatggcaaa?gcttccgagg?gagagaactt?cggtgatgac?aatagttcgg 660

gatcctattg?cgcgtgtgtt?aagcacttat?gaattttccg?tagaggtagc?agctaggttt 720

ttggtgcatc?ccaatttaac?ttctgcgtca?aggatgtcta?gccgcatacg?caagagtaat 780

gtaataagca?cactagacat?atggccatgg?aaatacctag?ttccatggat?gagagaagac 840

ttgtttgctc?ggcgagatgc?acgaaaattg?aaggaggtag?tgatcattga?ggacgataac 900

ccgtatgaca?tggaggagat?gcttatgcct?ttgcacaaat?atcttgatgc?gcctactgct 960

catgacatca?tccacaatgg?agcgactttt?cagattgcag?gattgacaaa?taactcccat 1020

ttatcagaag?cacacgaggt?tcggcattgt?gtgcagaaat?tcaaaagcct?tggtgagtct 1080

gttctccaag?ttgccaagag?gaggctagac?agcatgttgt?atgttggact?gacagaggag 1140

cacagggaat?ctgcatcact?ttttgccaat?gtagtgggtt?ctcaagtgct?gtctcaagtg 1200

gttccgtcca?atgcaactgc?gaaaatcaaa?gctcttaaat?cagaagcaag?tgtcacaatt 1260

tcagaaaccg?ggtcagataa?gagtaatatt?cagaatggta?catctgaagt?tacattgaat 1320

aaggcagaag?ctaagagtgg?gaatatgacg?gtaaaaaccc?ttatggaagt?ctatgaaggc 1380

tgcatcactc?atttacgaaa?gtcccaagga?accagacggg?tcaactctct?gaagagaata 1440

actccagcaa?attttacaag?agggacgcgt?acaagagttc?ctaaagaggt?cattcagcag 1500

atcaaatcgc?ttaacaacct?cgatgtggag?ctctacaaat?atgcaaaagt?aatctttgcc 1560

aaagaacatg?aattagtgtc?gaataagttg?atctcaagtt?ctaagagaag?cattgttgat 1620

ctgccgagtg?agttaaagag?cgtattggga?gaaatgggtg?aagagaagct?atggaagttc 1680

gtaccagtgg?cattgatgct?tttattgatc?gtcctcttct?ttctatttgt?aaacgctaaa 1740

aggagaagaa?cctccaaagt?taagatttga?tcattgtcac?tatattttgg?accaaatcag 1800

caatttgaag?aagcttttgg?ggtccaattt?gtaatattat?tctctttctt?tttctacttt 1860

tcgtggaaaa?agaaaaagtt?ttgggtgttg?ccttacctta?caggtgaaaa?agggaaaatc 1920

acaag 1925

<210>2

<211>4881

<212>DNA

＜213〉Arabidopis thaliana (Arabidopsis thaliana)

<400>2

gtgaagaaca?aaatattaca?caaagagggg?atgtatagtt?gtatacccac?ggtttaccga 60

tttcggtttt?cgttccgata?accggttatg?tatttcattt?tgtattatat?agaaggcatt 120

ggtcaaaccg?cactcgaacc?agaaaggaaa?gtgcattcat?actattactg?actagtggaa 180

aaaaaaaaaa?aaaaaaagca?ttggtctctc?tcaaaaaaga?ttaattcgtt?tggctaaatc 240

aaatttgaca?ctagtgtctt?cctcagtgcg?tgtttataat?ttttctgggt?ggcgagaaac 300

caagagaaaa?tctcgtgacc?ctgttctctc?gggagcgcac?acaatctctc?ttcttcttcg 360

tcttcctcct?ccaatcgatg?taatcacaca?cacacacaaa?aacacatctt?ttccttgttt 420

tcgtaggacc?tagtagatag?cgagttttgc?ccccaaaaca?gcgatcgagg?gaatttatga 480

gcacgagatc?tcgattcccc?atacacgtag?gcctctgctt?ctctctcttg?ttcctttgtt 540

cctttcttta?ttcgattttc?tgggtccctt?ttgccctgtt?tttcctctgt?ctccttcccc 600

caaaatcttt?cggttttgtt?tttatacaac?ttgcaaccgt?taatgttcat?cctcaccgat 660

tgatcaccta?ctcttttccc?ttcctgggtt?ttgtttcttt?cgaataaaga?ttgatccttc 720

tttcacttat?aatcgttttg?gtaatcagta?gggtttggaa?tcgaggcaca?atgcaaatga 780

actctgtttg?gaagctgtct?cttgggttat?tacttcttag?ctcaggtatg?tatgcagaat 840

ggttttacaa?aattgttcat?ttgataaaaa?agctggactt?gtctctgatg?ttcttggaat 900

aattggcaaa?ttatatctca?gttattggct?cttttgcgga?acttgatttt?ggccattgcg 960

aaactcttgt?gaaaaaatgg?gctgattctt?cttcatctcg?tgaagaacat?gttaataaag 1020

acaaacgctc?gcttaaggat?ttgctcttct?ttctccacgt?tccgcgaact?ggaggcagaa 1080

catattttca?ttggtgattt?gatttccttt?accaaaaagt?ttttgagaga?ttcttttatg 1140

attgttgtgt?ggttaaggct?gtgtttgtgc?tcttgtagtt?ttttgaggaa?gttgtatgat 1200

agctctgagg?aatgtcctcg?atcttacgac?aagctccact?tcaatccaag?gtatcttaat 1260

ttcatcagtg?ctctcgctaa?ttgtacgtct?ttatttgatg?ccaaagactc?tttcttgtgg 1320

aaacaataag?atggcttaca?gaatgctgtt?tgtacattct?attggttgtt?aagttgactt 1380

tagatagctt?agctagacat?atataaagtt?aaatattgtt?tcatatgaaa?atcaattttc 1440

cctagcctcc?tttacctaaa?gaaaaacaat?gtgaaaaaag?tattttgcct?gtggtgttac 1500

taccttcgga?cagttaagat?cgtgttgact?cgtaaattta?taggaaggaa?aagtgcaagt 1560

tgttagccac?acatgatgat?tatagtttga?tggcaaagct?tccgagggag?agaacttcgg 1620

tgatgacaat?agttcgggat?cctattgcgc?gtgtgttaag?cacttatgaa?ttttccgtag 1680

aggtagcagc?taggtttttg?gtgcatccca?atttaacttc?tgcgtcaagg?atgtctagcc 1740

gcatacgcaa?gagtaatgta?ataagcacac?tagacatatg?gccatggaaa?tacctagttc 1800

catggatgag?agaagacttg?tttgctcggg?tatgtcgacc?tatcccattc?gtcttttttg 1860

gcttttaagc?tagaacatga?taagaacaca?taaaacttgg?gctgaagctt?tttttactca 1920

tccttggctt?ttcttagcga?gatgcacgaa?aattgaagga?ggtagtgatc?attgaggacg 1980

ataacccgta?tgacatggag?gagatgctta?tgcctttgca?caaatatctt?gatgcgccta 2040

ctgctcatga?catcatccac?aatggagcga?cttttcaggt?actttttcat?gctatttgtt 2100

taagttttca?aagttattga?ttagagatct?ccagagcatt?ttcatttcac?ttcacagatt 2160

gcaggattga?caaataactc?ccatttatca?gaagcacacg?aggttcggca?ttgtgtgcag 2220

aaattcaaaa?gccttggtga?gtctgttctc?caagttgcca?aggtagtaac?acttcttctt 2280

catcatagtt?cactgcgtct?tggtattgtg?cttttaaaag?cagcagattc?tgtaagcttt 2340

tttaaatttt?gcagaggagg?ctagacagca?tgttgtatgt?tggactgaca?gaggagcaca 2400

gggaatctgc?atcacttttt?gccaatgtag?tgggttctca?agtgctgtct?caagtggttc 2460

cgtccaatgc?aactgcgaaa?atcaaagctc?ttaaatcagg?ttggtgtaag?tttcttaata 2520

ccatcgctgg?gctatctaag?gagaatctag?caacttagtt?gaaggacatg?gtgatcattt 2580

ttttagtcac?agtgtacagt?tttttcagag?taccgctttg?gagatggtat?ttactttagt 2640

ttttggcttt?tgaaagtcaa?aacccgtcat?attattcatt?aaaatgtttg?actgatattt 2700

ttcattcact?ttttaggagt?ttgattttta?attacctagg?gaagctactt?attgttctct 2760

gttatttggg?gattatcgta?taaggtttga?ttgtctttca?tcatgaagat?ttttctgatc 2820

tatgtttctt?gttgtcacat?catctttaaa?ggttctcttg?ccctcttttg?ttttagactt 2880

gtgtcttgtt?actctgggtc?cctgggttta?ttaaagtttc?tttgttggtt?aatcatagtc 2940

tcttagagta?agtagaatga?tgatatgaca?gttcatgtgc?atacagccta?ggctttctat 3000

tgcttttgtt?gtatttaaag?tcaggataac?aaatggttgt?acttgcactc?accaaattgt 3060

caactttatt?tccttgatgc?ttcagaagca?agtgtcacaa?tttcagaaac?cgggtcagat 3120

aagagtaata?ttcaggtaaa?gtcctgctgc?agaaggtaat?aatttgtttg?ttcaactgtc 3180

atttttgtaa?ttgtcttgct?gaaaatggct?gtgacattac?agaatggtac?atctgaagtt 3240

acattgaata?aggcagaagc?taagagtggg?aatgtaagta?gaatcccttt?tccaatttat 3300

tatcaacgct?tgagaccttg?tgtgactttt?atatatacac?ttcagatgac?ggtaaaaacc 3360

cttatggaag?tctatgaagg?ctgcatcact?catttacgaa?agtcccaagg?aaccagacgg 3420

gtcaactctc?tgaagagaat?aactccagca?aattttacaa?gaggggtaaa?gtgttttcgt 3480

acatgatcta?aacttagtaa?agttaatgag?agaaggctgg?aataatgaaa?tcttttgaca 3540

atgcatctaa?cagacgcgta?caagagttcc?taaagaggtc?attcagcaga?tcaaatcgct 3600

taacaacctc?gatgtggagc?tctacaaata?tgcaaaagta?atctttgcca?aagaacatga 3660

attagtgtcg?aataagttga?tctcaagtgt?aagaaacttc?ttgtcttcac?atttcgcctt 3720

ttagtttctg?tctgatataa?gactaaactc?catgattcgt?gtgcagtcta?agagaagcat 3780

tgttgatctg?ccgagtgagt?taaagagcgt?attgggagaa?atgggtgaag?agaagctatg 3840

gaagttcgta?ccagtggcat?tgatgctttt?attgatcgtc?ctcttctttc?tatttgtaaa 3900

cgctaaaagg?agaagaacct?ccaaagttaa?gatttgatca?ttgtcactat?attttggacc 3960

aaatcagcaa?tttgaagaag?cttttggggt?ccaatttgta?atattattct?ctttcttttt 4020

ctacttttcg?tggaaaaaga?aaaagttttg?ggtgttgcct?taccttacag?gtgaaaaagg 4080

gaaaatcaca?agtaaaaaaa?aactctctat?ctactttatt?ttaaaaggct?aaacaaaaca 4140

ggtaaagtca?caaaattagt?gcttcttttt?ttttttttgt?ttaagtgagc?ttaatcaata 4200

aagataaaag?taataagatt?tcacaatttt?tttttttttt?tttttttgag?aaagtacact 4260

actaactggt?ttgaatttga?ctgtaccatt?tttcacgcaa?cagactgtgt?gcaatttaaa?4320

gtacagttca?tacaaatctc?tataaaacaa?attgtacaaa?agcagtaaat?atacccccac?4380

gtgatatttg?ttttcttatc?aacgacgcag?tgtaaacaat?aatatgtcaa?aaattttgaa?4440

gaaaaaatca?tataaactat?ataagaagaa?aaaatgaaaa?gaaatatgaa?gatcgtgtat?4500

attaattttt?ttttatataa?aaaatactaa?taataataat?tgattgatgg?gaggaagata?4560

gtaagtgaaa?gggatgaaac?atatcatgtg?ggagatggaa?gcaaagagtt?aggcaaggga?4620

cccactttcc?catgaggaaa?cgccataatc?atctttatta?ttattattat?ttcggttctc?4680

gtctcttaat?ccttcgtcgg?tgatctctcc?atttcacttt?gactctctct?attatactaa?4740

aggtttttag?tatattatgt?taaacccaac?ttatttgttt?gctacgtaaa?ttaaattata?4800

catcaattca?tgatactatc?tattattatt?gctagagatc?gagatctatc?aaccatcctt?4860

ccctcaaggt?tattttgcca?t 4881

<210>3

<211>500

<212>PRT

＜213〉Arabidopis thaliana (Arabidopsis thaliana)

<400>3

Met?Gln?Met?Asn?Ser?Val?Trp?Lys?Leu?Ser?Leu?Gly?Leu?Leu?Leu?Leu

1 5 10 15

Ser?Ser?Val?Ile?Gly?Ser?Phe?Ala?Glu?Leu?Asp?Phe?Gly?His?Cys?Glu

20 25 30

Thr?Leu?Val?Lys?Lys?Trp?Ala?Asp?Ser?Ser?Ser?Ser?Arg?Glu?Glu?His

35 40 45

Val?Asn?Lys?Asp?Lys?Arg?Ser?Leu?Lys?Asp?Leu?Leu?Phe?Phe?Leu?His

50 55 60

Val?Pro?Arg?Thr?Gly?Gly?Arg?Thr?Tyr?Phe?His?Cys?Phe?Leu?Arg?Lys

65 70 75 80

Leu?Tyr?Asp?Ser?Ser?Glu?Glu?Cys?Pro?Arg?Ser?Tyr?Asp?Lys?Leu?His

85 90 95

Phe?Asn?Pro?Arg?Lys?Glu?Lys?Cys?Lys?Leu?Leu?Ala?Thr?His?Asp?Asp

100 105 110

Tyr?Ser?Leu?Met?Ala?Lys?Leu?Pro?Arg?Glu?Arg?Thr?Ser?Val?Met?Thr

115 120 125

Ile?Val?Arg?Asp?Pro?Ile?Ala?Arg?Val?Leu?Ser?Thr?Tyr?Glu?Phe?Ser

130 135 140

Val?Glu?Val?Ala?Ala?Arg?Phe?Leu?Val?His?Pro?Asn?Leu?Thr?Ser?Ala

145 150 155 160

Ser?Arg?Met?Ser?Ser?Arg?Ile?Arg?Lys?Ser?Asn?Val?Ile?Ser?Thr?Leu

165 170 175

Asp?Ile?Trp?Pro?Trp?Lys?Tyr?Leu?Val?Pro?Trp?Met?Arg?Glu?Asp?Leu

180 185 190

Phe?Ala?Arg?Arg?Asp?Ala?Arg?Lys?Leu?Lys?Glu?Val?Val?Ile?Ile?Glu

195 200 205

Asp?Asp?Asn?Pro?Tyr?Asp?Met?Glu?Glu?Met?Leu?Met?Pro?Leu?His?Lys

210 215 220

Tyr?Leu?Asp?Ala?Pro?Thr?Ala?His?Asp?Ile?Ile?His?Asn?Gly?Ala?Thr

225 230 235 240

Phe?Gln?Ile?Ala?Gly?Leu?Thr?Asn?Asn?Ser?His?Leu?Ser?Glu?Ala?His

245 250 255

Glu?Val?Arg?His?Cys?Val?Gln?Lys?Phe?Lys?Ser?Leu?Gly?Glu?Ser?Val

260 265 270

Leu?Gln?Val?Ala?Lys?Arg?Arg?Leu?Asp?Ser?Met?Leu?Tyr?Val?Gly?Leu

275 280 285

Thr?Glu?Glu?His?Arg?Glu?Ser?Ala?Ser?Leu?Phe?Ala?Asn?Val?Val?Gly

290 295 300

Ser?Gln?Val?Leu?Ser?Gln?Val?Val?Pro?Ser?Asn?Ala?Thr?Ala?Lys?Ile

305 310 315 320

Lys?Ala?Leu?Lys?Ser?Glu?Ala?Ser?Val?Thr?Ile?Ser?Glu?Thr?Gly?Ser

325 330 335

Asp?Lys?Ser?Asn?Ile?Gln?Asn?Gly?Thr?Ser?Glu?Val?Thr?Leu?Asn?Lys

340 345 350

Ala?Glu?Ala?Lys?Ser?Gly?Asn?Met?Thr?Val?Lys?Thr?Leu?Met?Glu?Val

355 360 365

Tyr?Glu?Gly?Cys?Ile?Thr?His?Leu?Arg?Lys?Ser?Gln?Gly?Thr?Arg?Arg

370 375 380

Val?Asn?Ser?Leu?Lys?Arg?Ile?Thr?Pro?Ala?Asn?Phe?Thr?Arg?Gly?Thr

385 390 395 400

Arg?Thr?Arg?Val?Pro?Lys?Glu?Val?Ile?Gln?Gln?Ile?Lys?Ser?Leu?Asn

405 410 415

Asn?Leu?Asp?Val?Glu?Leu?Tyr?Lys?Tyr?Ala?Lys?Val?Ile?Phe?Ala?Lys

420 425 430

Glu?His?Glu?Leu?Val?Ser?Asn?Lys?Leu?Ile?Ser?Ser?Ser?Lys?Arg?Ser

435 440 445

Ile?Val?Asp?Leu?Pro?Ser?Glu?Leu?Lys?Ser?Val?Leu?Gly?Glu?Met?Gly

450 455 460

Glu?Glu?Lys?Leu?Trp?Lys?Phe?Val?Pro?Val?Ala?Leu?Met?Leu?Leu?Leu

465 470 475 480

Ile?Val?Leu?Phe?Phe?Leu?Phe?Val?Asn?Ala?Lys?Arg?Arg?Arg?Thr?Ser

485 490 495

Lys?Val?Lys?Ile

500

Claims (9)

1, a kind of associated protein for plant root growth is the protein with one of following amino acid residue sequences:

1) amino acid residue sequence of the SEQ ID № .3 in the sequence table;

2) with the SEQ ID № .3 amino acid residue sequence in the sequence table through the replacement of one or several amino-acid residue and/or disappearance and/or interpolation and have the active protein of associated protein for plant root growth.

2, the encoding gene of the described associated protein for plant root growth of claim 1, the 5 ' end 268-1770 position nucleotide sequence with sequence 2 in sequence table.

3, encoding gene according to claim 2 is characterized in that: the cDNA gene of described associated protein for plant root growth has one of following nucleotide sequence:

1) SEQ ID № in the sequence table: 2 nucleotide sequence;

2) SEQ ID № in the code sequence tabulation: the polynucleotide of 3 protein sequences;

3) under the rigorous condition of height can with SEQ ID № in the sequence table: the nucleotide sequence of the 2 dna sequence dnas hybridization that limit.

4, encoding gene according to claim 3 is characterized in that: the genomic gene of described associated protein for plant root growth has one of following nucleotide sequence:

1) SEQ ID № in the sequence table: 1 nucleotide sequence;

2) SEQ ID № in the code sequence tabulation: the polynucleotide of 3 protein sequences;

3) under the rigorous condition of height can with SEQ ID № in the sequence table: the nucleotide sequence of the 1 dna sequence dna hybridization that limits.

5, the recombinant expression vector that contains any described associated protein for plant root growth encoding gene among the claim 2-4.

6, the transgenic cell line that contains any described associated protein for plant root growth encoding gene among the claim 2-4.

7, the host bacterium that contains any described associated protein for plant root growth encoding gene among the claim 2-4.

8, the application of any described associated protein for plant root growth encoding gene in promoting plant root growth among the claim 2-4.

9, application according to claim 8 is characterized in that: described plant is an Arabidopis thaliana.

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