CN113667679A

CN113667679A - PsIAA27 gene fragment and application thereof

Info

Publication number: CN113667679A
Application number: CN202110771464.2A
Authority: CN
Inventors: 王政; 何松林; 孟新亚; 石力匀; 贺丹; 尚文倩; 娄雪源; 申玉晓; 宋莹莹; 徐玉凤
Original assignee: Henan Agricultural University
Current assignee: Henan Agricultural University
Priority date: 2021-07-08
Filing date: 2021-07-08
Publication date: 2021-11-19
Anticipated expiration: 2041-07-08
Also published as: CN113667679B

Abstract

The invention relates to a PsIAA27 gene fragment and application thereof. The PsIAA27 gene fragment comprises a nucleotide sequence shown in SEQ I D NO. 2. The PsIAA27 gene fragment expresses the Ps IAA27 protein. The PsIAA27 gene fragment, and expression vectors comprising the PsIAA27 gene fragment, may be used to promote plant rooting. The invention has the advantages of being beneficial to the elongation of the plant root system, the germination of lateral roots and the increase of the number of the lateral roots.

Description

PsIAA27 gene fragment and application thereof

Technical Field

The invention belongs to the field of plant physiology and molecular biology application, and particularly relates to a PsIAA27 gene fragment and application thereof.

Background

The root is one of the six major organs of the plant, is the vegetative organ of the plant, is usually located under the ground surface, is responsible for absorbing the water content of the soil and dissolving the inorganic salts therein, and has the function of supporting, reproducing, and storing synthetic organic substances. Therefore, the growth conditions of roots have an important role in the growth of plants. However, in the prior art, the knowledge of genes capable of promoting the lateral root germination of a plant root system and increasing the number of the lateral roots is limited, and whether new genes can be found and used for the growth of the root system is a problem to be solved urgently.

Peony (Paeonia suffruticosa Andr.) is deciduous shrub of Paeonia of Paeoniaceae, also called Paeonia lactiflora, Fugui flower, and the like, is a traditional famous flower in China, and has high ornamental value and medicinal value. The peony has various varieties, and the flower is large and bright, graceful and noble, and is widely advocated as the national flower of China. The conventional propagation of the peony mainly takes grafting as a main part, but the propagation period is long and the propagation rate is low, so that the industrialized production of the peony seedlings is greatly restricted. Tissue culture has the advantages of high propagation speed, high coefficient, short period, capability of keeping excellent characters of a mother plant and the like, and is widely applied to rapid propagation of plants in recent years. However, the peony test-tube plantlet has the problems of high adventitious root induction difficulty, low rooting rate, poor adventitious root quality, low transplanting survival rate and the like, and the industrial development of the peony is seriously influenced. Therefore, the research on the root system generation mechanism has important significance for peony tissue culture.

The auxin is used as a main hormone for promoting the generation of adventitious roots, and not only has a direct effect on the regulation and control of the growth and development of plant root systems, but also can indirectly regulate and control the expression of genes related to the growth and development of the root systems.

Disclosure of Invention

In view of the problems in the prior art, the invention provides a PsIAA27 gene fragment and application thereof. The invention has the advantages of being beneficial to the elongation of the plant root system, the germination of lateral roots and the increase of the number of the lateral roots.

The technical scheme for solving the technical problems is as follows:

the invention provides a PsIAA27 gene fragment, wherein the nucleotide sequence of the PsIAA27 gene fragment comprises the nucleotide sequence shown in SEQ ID NO. 2.

The invention provides a PsIAA27 protein, and the PsIAA27 protein comprises the protein expressed by the PsIAA27 gene segment.

Further, the amino acid sequence of the PsIAA27 protein includes the amino acid sequence shown in SEQ ID NO. 3.

The invention provides a group of primers for amplifying PsIAA27 gene fragments, which comprise a nucleotide sequence shown in SEQ ID NO.8 and a nucleotide sequence shown in SEQ ID NO. 9. The primers can quickly and accurately amplify the PsIAA27 gene fragment.

The invention provides an expression vector, which comprises the PsIAA27 gene segment.

The invention provides a strain comprising a PsIAA27 gene fragment, wherein the strain comprises the expression vector.

The PsIAA27 gene fragment or the expression vector can be applied to promoting plant rooting.

Further, the PsIAA27 gene fragment or the expression vector is applied to promoting the growth and/or germination of lateral roots of plants.

The invention provides a method for promoting plant rooting, which comprises the following steps: the PsIAA27 gene was expressed in plants. Preferably, the PsIAA27 gene is overexpressed in a plasmid or other form.

Drawings

FIG. 1 is a graph of the predicted results of the protein domain of PsIAA27.

FIG. 2 is a diagram showing the comparison of the sequences of the amino acids encoded by the PsIAA27 gene and the similar histone proteins.

FIG. 3 is a graph showing the results of expression levels of PsIAA27 gene at different key rooting stages.

FIG. 4 is a diagram showing the results of verifying the plasmid pD1301S-PsIAA 27.

FIG. 5 is a graph showing the results of the expression level of the PsIAA27 gene of transgenic Arabidopsis thaliana T1 generation.

FIG. 6 is a graph showing the results of root length and root number expression of T1 generations of Arabidopsis thaliana transformed with PsIAA27 gene.

FIG. 7 is a graph showing the results of observation of representatives of Arabidopsis thaliana T1 transgenic for PsIAA27 gene, A: a control CK; b: transgenic Arabidopsis thaliana with PsIAA27 gene.

Detailed Description

The principles and features of this invention are described below in conjunction with the following drawings, which are set forth by way of illustration only and are not intended to limit the scope of the invention.

The invention obtains an auxin gene PsIAA27 related to peony rooting regulation and control and carries out RT-qPCR expression and functional verification. The regulation and control effect of the method on the adventitious root differentiation of the peony is disclosed, and theoretical guidance is provided for discussing the rooting mechanism of the peony and establishing a tissue culture rapid propagation system.

The following description is given by way of specific examples.

Example 1

1. qRT-PCR and clone analysis of rooting differential gene

Screening and obtaining 1 gene with obvious expression difference according to the result of transcriptome data of the peony root system in different periods: unigene13430_ All. Primer design is carried out by using Primer 5.0, synthesis is carried out by entrusting Guangzhou Secho biotech GmbH, the peony beta-Tubulin gene is selected as an internal reference gene (EF608942) (Meshkexie, 2013), PCR amplification of Unigene13430_ All and the internal reference gene is respectively carried out under the same amplification condition, and related gene ID and Primer sequence are shown in Table 1.

TABLE 1 primers for qRT-PCR validation

PCR amplification reaction System:

PCR amplification method

The gene sequence (the nucleotide sequence is shown as SEQ ID NO. 1) of the Unigene13430_ All is successfully obtained by taking root system cDNA of the peony at different periods as a template and adopting the PCR method to carry out PCR amplification. The total length of CDS is 843bp (the nucleotide sequence is shown in SEQ ID NO.2), 280 amino acids are coded (the amino acid sequence is shown in SEQ ID NO. 3). In combination with the sequence analysis of Smart-Blast protein, the Unigene13430_ All gene has a complete AUX-IAA domain, which is determined to be an IAA gene family gene (FIG. 1). The sequence alignment result shows that the protein has higher similarity (figure 2) with the IAA27 protein amino acid sequences of grapes, walnuts, plum blossom and jujubes, and the similarity is 88 percent, so the peony PsIAA27 gene is named.

2. Construction of Arabidopsis thaliana overexpression vector

Primers containing enzyme cutting sites are designed, as shown in the following table 2, plasmids containing full-length genes (the full-length genes refer to SEQ ID NO.2) are used as templates, complete reading frames of the genes are amplified respectively, PCR products are recovered and connected into pGEM-T easy vectors, escherichia coli is transformed, single clones are selected for sequencing, and plasmids are extracted correctly through sequencing.

The plasmid extraction method is as follows:

1) selecting the transformed escherichia coli positive single colony on a fresh 5mL LB liquid culture medium, and culturing overnight at 37 ℃ and 150-;

2) subculturing into LB liquid culture medium at a ratio of 1:100, and culturing to logarithmic phase (OD600 of 0.4-0.6);

3) subpackaging the bacterial liquid into 50mL centrifuge tubes, and standing on ice for 10-20 min;

4) centrifuging at 4 deg.C and 4500g for 10min, quickly removing supernatant, sterilizing with 1/5 volume precooled sterilized 100mmol/L CaCl₂Weighing cells, collecting the heavy suspension in the same centrifuge tube, and standing on ice for 30 min;

5) centrifugation at 4500g for 10min at 4 deg.C, careful decanting of the supernatant, and gentle resuspension of the cells in pre-cooled sterilized 100mmol/L CaCl2+ 15% glycerol (concentration factor 100-₂+ 15% glycerol);

6) subpackaging 100 μ L/tube into sterile 1.5mL centrifuge tube, quick freezing in liquid nitrogen, and storing at-80 deg.C.

Transformation and recombinant clone screening: the method is carried out according to the conventional method.

7) Selecting positive clone Escherichia coli, then implanting into ampicillin resistant liquid LB culture medium with final concentration of 100 mug/mL, and culturing environment: the temperature is 37 ℃, 5-8 hours, and finally the sequencing is sent to a gene company for sequencing.

8) Plasmids were extracted using a plasmid miniprep kit (Tiangen), and the specific procedures were performed according to the instructions.

TABLE 2 primers used for construction of overexpression vectors

3. Rooting difference related gene qRT-PCR

And the expression quantity of the PsIAA27 gene is analyzed in the key rooting period of the cutting seedlings and the grafted seedlings by real-time fluorescent quantitative PCR. FIG. 3 is an experimental result of expression of PsIAA27 gene in different rooting key stages, wherein ZC represents growth of root primordium, MF represents germination of root primordium, PD represents expansion of root primordium, and each group comprises seedlings, cuttage and grafting in sequence from left to right. As shown in fig. 3, PsIAA27 showed significant differences in the different rooting periods of the cuttings and the grafted seedlings. The expression quantity in the seedling is increased and then decreased, the expression quantity is opposite to that in the cutting seedling and the grafting seedling, and the expression abundance in the cutting seedling is higher than that in the grafting seedling. As can be shown in fig. 3, the expression level of the PsIAA27 gene is high in the root primordium germination stage of seedlings, the expansion expression abundance of the root primordium of cutting seedlings and grafted seedlings is high, and the expression level of the PsIAA27 gene is highest in the cutting seedlings.

The real-time fluorescent quantitative PCR method comprises the following steps:

qRT-PCR reaction was carried out using TAKARA kit, and PCR reaction solution was prepared in the following order of reagents.

In the PCR reaction solution, the forward primer can be PsIAA27.RT.F1, and the reverse primer can be PsIAA27. RT.R1; the cDNA solution can be cDNA of roots of seedlings in different rooting periods, and is obtained by using an RNA extraction and reverse transcription kit of a Tiangen organism.

By using

The method of (3) is used for PCR amplification, and the reaction procedure is as follows:

relative expression amount of Gene 2^-ΔΔCt[298]The results were expressed and analyzed by Microsoft Excel.

4. Construction of Arabidopsis thaliana overexpression vector

The target gene (shown in SEQ ID NO.2) was digested simultaneously with ScaI (11267) and SalI (11236). Synthesizing genes on a pUC57 vector, taking bacterial liquid containing synthetic genes, shaking bacteria to extract plasmids, carrying out double enzyme digestion by ScaI and SalI, simultaneously carrying out enzyme digestion on pD1301S by the same enzyme, connecting recovered products by enzyme T4, transferring the connected products to DH5a competent cells, culturing at 37 ℃, selecting grown colonies for purification, finally picking bacteria to carry out shaking bacteria to extract plasmids, and obtaining a final vector pD1301S-PsIAA 27. FIG. 4 shows the results of the verification of plasmid pD1301S-PsIAA27, samples 1-9 of pD1301S-PsIAA27, and DNA marker in the left column with no reference numeral, and it can be seen from FIG. 4 that the sizes of the amplified

bands

1, 2 and 3 are substantially the same as PsARF, and it can be confirmed that the recombinant plasmid is correct.

DH5a competent cells (purchased from the Tokyo Union) were maintained at the plant physiology laboratory of the college of forestry, university of agriculture, Henan.

The pUC57 vector (purchased from the Pimenta Subco) involved in the above steps was then stored in the plant physiology laboratory of the forestry institute of agriculture university, Henan, and the information of pUC57 vector was referred to the Pimenta Subco official network, and the public could obtain the pUC57 vector from the laboratory and repeat the steps of the present invention for non-commercial purposes.

5. Transformation of Arabidopsis thaliana for functional verification

Transforming agrobacterium LBA4404 with the constructed super expression vector plasmid (pD1301S-PsARF) (the method is introduced in the super expression vector construction part) to obtain agrobacterium containing the target gene vector (LBA4404-PsARF), and respectively inoculating the agrobacterium containing the target gene vector (LBA4404-PsARF) and the agrobacterium containing the empty vector in 10mL YEB liquid culture medium (containing 50 mg. L)^-1Rif +50mg·L^-1Kan), 28 ℃, 200rpm, and shaking for 12-14 h.

5mL of this medium was added to 200mL of YEB liquid medium (containing 50 mg. multidot.L)^-1Rif +50 mg. L of rifampicin^-1Kanamycin Kan), 28 ℃, 200rpm, shaking culture for 6-12h (to OD)₆₀₀＝0.8-1.0)；

YEB liquid medium formula: 0.5g of peptone, 0.1g of yeast powder, 0.5g of beef extract, 0.5g of sucrose and 0.05g of magnesium sulfate heptahydrate, and the components are dissolved in 100ml of distilled water;

centrifuging at 4000rpm for 10min, collecting thallus, and adding 200mL MS salt solution (containing 1/2MS, 5% sucrose, 200. mu.L. L)^-1Silwet L-77，1mL·L^-1Triton X-100) resuspending;

MS formula: NH (NH)₄NO₃ 1650mg·L^-1、KNO₃ 1900mg·L^-1、CaCl₂·2H₂O 440 mg·L^-1、MgSO₄·7H₂O 370mg·L^-1、KH₂PO₄ 170mg·L^-1、KI 0.83mg·L^-1、H₃BO₃ 6.2mg·L^-1、MnSO₄·4H₂O 22.3mg·L^-1、ZnSO₄·7H₂O 8.6mg·L^-1、Na₂MoO₄·2H₂O 0.25mg·L^-1、CuSO₄·5H₂O 0.025mg·L^-1、CoCl₂·6H₂O 0.025mg·L^-1、FeSO₄·7H₂O 27.8mg·L^-1、Na₂-EDTA·2H₂O 37.3mg·L^-1Inositol 100 mg.L^-10.5 mg. L of nicotinic acid^-1Vitamin B60.5mg. L^-1Vitamin B10.1mg. L^-12.0 mg. L of glycine^-1。

The reagents used in the above experiments were purchased from loranthus biotechnology limited.

When a large amount of wild arabidopsis plants bloom, the plants can be used for transformation, and the plants are watered fully 1d before transformation. And (3) soaking the arabidopsis inflorescence in the agrobacterium tumefaciens heavy suspension for about 1min, laterally placing a flowerpot, and covering the flowerpot with a freshness protection package for preserving moisture for 1 d. And 2d, taking the plants out of the freshness protection bags, placing the plants on the illumination culture rack after the plants are placed in the dark for 1d, and culturing the plants to be fruitful by a conventional method until mature T is harvested₀And (5) seed generation. Mature T₀Sowing seeds, conventionally culturing seedlings to obtain T1 generation plants, extracting RNA, reversely transcribing the RNA into cDNA, and performing related gene expression analysis and phenotype observation by using GAP as an internal reference gene.

Method for extracting RNA: RNA was extracted using a kit for RNA extraction in the Hua Yun Yang, and the specific procedures were carried out according to the instructions.

Method for reverse transcription into cDNA: the cDNA is synthesized by reverse transcription with RNA reverse transcription kit (Optimalaceae), and the specific operation is performed according to the instruction.

Agrobacterium (purchased from the Jiang Union organism) was maintained in the plant physiology laboratory of the forest institute of Henan university of agriculture.

6. Expression analysis of transgenic Arabidopsis thaliana T1 generation rooting related gene

FIG. 5 shows the results of experiments on the expression level of PsIAA27 gene in transgenic Arabidopsis T1 generation, wherein CK group is non-transgenic Arabidopsis and PsIAA27 group is overexpression transgenic Arabidopsis with PsIAA27 gene. As can be seen from FIG. 5, the expression level of the transgenic Arabidopsis T1 generation rooting gene PsIAA27 is significantly higher than that of the control, which indicates that the gene PsIAA27 is successfully transferred into Arabidopsis T1 generation, and the next step of verification can be carried out.

7. Transgenic Arabidopsis thaliana T1 representational type observations

FIG. 6 shows the experimental results of root length and root number expression of T1 generations of Arabidopsis thaliana transformed with PsIAA27 gene, CK group is non-transgenic Arabidopsis thaliana, PsIAA27 group is Arabidopsis thaliana transformed with PsIAA27 gene, the left figure shows the experimental results of root length, and the right figure shows the experimental results of root number.

FIG. 7 Experimental results of observations of representatives of the Arabidopsis thaliana T1 transgenic for PsIAA27 gene, A: control CK, i.e. non-transgenic arabidopsis; b: transgenic Arabidopsis thaliana with PsIAA27 gene.

FIGS. 6 and 7 show the growth of transgenic Arabidopsis T1 generation, with transgenic PsIAA27 Arabidopsis plants significantly better than controls in root length and with an increase in root number compared to controls. The gene PsIAA27 was shown to promote root elongation and lateral root germination.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Sequence listing

<110> Henan university of agriculture

<120> PsIAA27 gene fragment and application thereof

<141> 2021-07-08

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tatgaccgta gtttttgaaa ggtcgccggg aactttccat cacttgttgg gagctgttcg 60

gaaaaggtga atgaaaggag aacaaagtag aaggggtaat atcgaaaaat aaaatcccaa 120

gctcaaacag tttcggttcc ttctctactt tacaaatagc aagtccttcc attattagat 180

tctgaataag aacatttcca tcatcttatt agtttcagta gtgtgaaaga ttatcctttt 240

ttatcaactc ttatctttct tcttcttctt cttcttcgta accattccat gctttattgt 300

acacaacata taaaaccctc gttgaaattt gatgtctacg ccactagaac atgattacat 360

aggcttatca ggggtttcct caatggagag ctccgaaaag atttccacta ctacatcctc 420

tgacaacatc gctgaaacga acaatgttct taatttaaag gccactgagt tgagacttgg 480

actccctggt tcagagtctc ctgaaagaag tgataaatct gttgaggcat tggaggataa 540

taagaaatcg taccctcttg gggtactcaa gaacttggtc tctggtgcca agcggggctt 600

ctctgacgct atcgataatg gttctgggaa ctgggttttc tctggggttg gtggatctga 660

gtctgattcg tcaaaaaatt gtggtttgtt ctctcccaga ggtggacatg ctgggaagcc 720

tattggtggg tcggatggta acactcagca gctgggtttg gttgctccga ccgacttcgc 780

gcctcagtca ccaaagccag tgcaggagaa ggcgcctcag ttttctgcta caaatggcca 840

tgggattagg cctgctgcaa aggcacaggt agtaggatgg ccaccaattc gctctttccg 900

gaaaaatacg atggctaata agactccaaa gaacgacgat gattcagagg gccagttggg 960

gtcgggttgt atttatgtca aagttagtat ggacggtgct ccatacctta ggaaagttga 1020

tatcaaaacc tattgcagct atatggaact atcttcagca ctggaaaaga tgtttagctc 1080

agttggtaag cttctatgca tcctatcatc ctcggtctgt gttaagttgc tatgcaattg 1140

tttatctttc tcttgcttgt gcacagacat ttgatttttg aagaaaactt cagaaagtcc 1200

ttgatccatg gtctgcacta tgcagagcca agtctgtgtt tctgactaca gtaattggaa 1260

gcagacaata agtgacaaat ctgattttca tatgcttcgt gaaagtatat cactcattta 1320

tgtaaaacag ggcatctgaa cacagaagtt ggttgtgcat aaactaccaa ataatataaa 1380

aatgcagcca tttatcgaat cctaggatac attattgata ttactaataa ggtgcaatta 1440

gagcatactt aatggtgagg agcaaaccca ttttccccat caatttggtg gctgatggtg 1500

ttttgccatc cccaacaggc actttgcacc tcctgcaaaa taccattaac ggcaaaaagg 1560

agggtttcca tgccaaatta cccctccctc cccttcaaat gccgtgcacg gcaaaagtgc 1620

cacctaggca attttttttg tcttttttcc ttttcatatt ttttgcctta atttaatatc 1680

tattatttta aattaataat tattttgatt tatacaatca ccatatttaa tattaaagtg 1740

tgtttgctac cttcttgttg gggatgaaaa ttacatatac accaaattgg gttcacaaaa 1800

tttgttacca taccaccaaa tagagtttgc taccaccgat tgctaccttc ttgttggaca 1860

tgctctaacc attcaaacac atttgaccca ctttaattcc tatgaagatc ataacagact 1920

cattaagttt ccatgcgttg tccaaatgtt tctttggtaa atttgacatc agcttatatg 1980

taaccgaaga gaaatggatt gttgttgctc cataaatgaa attttaaata aagataccct 2040

tttttcagaa attaacttta gtagcagaac atttttggtt actttgatta tataatatta 2100

tcgtatataa tacaaaatgt taaactacaa ttcaattctg gatatttgta ggatccttcc 2160

tctttctaaa acacagttta cgtttgtatt atggtaaaaa aactgaaatc ttagtgccat 2220

ttctcctcct gttttcatgt gtgtgtgtgt gtgtgtgcat gcgtgcgtgt gtctgtgtgt 2280

gttgggggag gggttagaga attgatacct atgcatacta caaacatagg aaatgtactt 2340

ataagatagt gatcacaaat catcattatg atagctttta aatatgatgt gtaaatcatg 2400

caaaatctct actactaggt tctggattaa gttgttcata ggaaactgca aacataggaa 2460

gtgtacttgt aagatagtgg tcataaatca ttctcatgat agcttttaaa tatgatctat 2520

aaatgaggca aaatctctac ttctaggttc ttgattaaga agttcatagg aaactgcccc 2580

cactgcatcc acccacccac acacacacag ggtaccgtga aatggatcct gagtatcatg 2640

gtatttttgg gacccagtac tctaaaatca acacaccaag tatttttttt cccccttata 2700

atgtatattg ttgcaaaaaa gtatgaagat acatatagag aagggaaaag acaaacagaa 2760

ttttggtaca ttttagggtt tcagttcatc tataattaca tatagtagca aaataatgta 2820

gcgaccaaat tgcatgccaa aataatgtag caaaagtagc ctactcctgg gtattgcgta 2880

tcggccgaag aagcgcacca tgtaccagta tccataccat ataccaaagg aaacagcata 2940

cagcattcca ggtaactatg gttgtatact tgtgttgtat cagttacaaa ctatgatgtg 3000

cagaatttca tttttaccga attttcttcg atgcccttct cgggaagttt ttattgttta 3060

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tattccctct caatcgaaat agtatatata tatatatata tatggctata tattttatca 3180

tttttttgtt tattgcagac ggtttcatct gaatatttat ggtgagcagt aatgccaata 3240

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<213> Artificial Sequence

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atgtctacgc cactagaaca tgattacata ggcttatcag gggtttcctc aatggagagc 60

tccgaaaaga tttccactac tacatcctct gacaacatcg ctgaaacgaa caatgttctt 120

aatttaaagg ccactgagtt gagacttgga ctccctggtt cagagtctcc tgaaagaagt 180

gataaatctg ttgaggcatt ggaggataat aagaaatcgt accctcttgg ggtactcaag 240

aacttggtct ctggtgccaa gcggggcttc tctgacgcta tcgataatgg ttctgggaac 300

tgggttttct ctggggttgg tggatctgag tctgattcgt caaaaaattg tggtttgttc 360

tctcccagag gtggacatgc tgggaagcct attggtgggt cggatggtaa cactcagcag 420

ctgggtttgg ttgctccgac cgacttcgcg cctcagtcac caaagccagt gcaggagaag 480

gcgcctcagt tttctgctac aaatggccat gggattaggc ctgctgcaaa ggcacaggta 540

gtaggatggc caccaattcg ctctttccgg aaaaatacga tggctaataa gactccaaag 600

aacgacgatg attcagaggg ccagttgggg tcgggttgta tttatgtcaa agttagtatg 660

gacggtgctc cataccttag gaaagttgat atcaaaacct attgcagcta tatggaacta 720

tcttcagcac tggaaaagat gtttagctca gttggtaagc ttctatgcat cctatcatcc 780

tcggtctgtg ttaagttgct atgcaattgt ttatctttct cttgcttgtg cacagacatt 840

tga 843

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<213> Artificial Sequence

<400> 3

Met Ser Thr Pro Leu Gly His Ala Thr Ile Gly Leu Ser Gly Val Ser

1 5 10 15

Ser Met Gly Ser Ser Gly Leu Ile Ser Thr Thr Thr Ser Ser Ala Ala

20 25 30

Ile Ala Gly Thr Ala Ala Val Leu Ala Leu Leu Ala Thr Gly Leu Ala

35 40 45

Leu Gly Leu Pro Gly Ser Gly Ser Pro Gly Ala Ser Ala Leu Ser Val

50 55 60

Gly Ala Leu Gly Ala Ala Leu Leu Ser Thr Pro Leu Gly Val Leu Leu

65 70 75 80

Ala Leu Val Ser Gly Ala Leu Ala Gly Pro Ser Ala Ala Ile Ala Ala

85 90 95

Gly Ser Gly Ala Thr Val Pro Ser Gly Val Gly Gly Ser Gly Ser Ala

100 105 110

Ser Ser Leu Ala Cys Gly Leu Pro Ser Pro Ala Gly Gly His Ala Gly

115 120 125

Leu Pro Ile Gly Gly Ser Ala Gly Ala Thr Gly Gly Leu Gly Leu Val

130 135 140

Ala Pro Thr Ala Pro Ala Pro Gly Ser Pro Leu Pro Val Gly Gly Leu

145 150 155 160

Ala Pro Gly Pro Ser Ala Thr Ala Gly His Gly Ile Ala Pro Ala Ala

165 170 175

Leu Ala Gly Val Val Gly Thr Pro Pro Ile Ala Ser Pro Ala Leu Ala

180 185 190

Thr Met Ala Ala Leu Thr Pro Leu Ala Ala Ala Ala Ser Gly Gly Gly

195 200 205

Leu Gly Ser Gly Cys Ile Thr Val Leu Val Ser Met Ala Gly Ala Pro

210 215 220

Thr Leu Ala Leu Val Ala Ile Leu Thr Thr Cys Ser Thr Met Gly Leu

225 230 235 240

Ser Ser Ala Leu Gly Leu Met Pro Ser Ser Val Gly Leu Leu Leu Cys

245 250 255

Ile Leu Ser Ser Ser Val Cys Val Leu Leu Leu Cys Ala Cys Leu Ser

260 265 270

Pro Ser Cys Leu Cys Thr Ala Ile

275 280

<210> 4

<211> 24

<212> DNA

<213> Artificial Sequence

<400> 4

aaagtagcct actcctgggt attg 24

<210> 5

<211> 25

<212> DNA

<213> Artificial Sequence

<400> 5

ccatagttac ctggaatgct gtatg 25

<210> 6

<211> 24

<212> DNA

<213> Artificial Sequence

<400> 6

tgagcaccaa agaagtggac gaac 24

<210> 7

<211> 23

<212> DNA

<213> Artificial Sequence

<400> 7

cacacgcctg aacatctcct gaa 23

<210> 8

<211> 22

<212> DNA

<213> Artificial Sequence

<400> 8

acccagtact ctaaaatcaa ca 22

<210> 9

<211> 20

<212> DNA

<213> Artificial Sequence

<400> 9

tgaactgaaa ccctaaaatg 20

Claims

1. A PsIAA27 gene fragment, wherein the nucleotide sequence of the PsIAA27 gene fragment comprises the nucleotide sequence shown in SEQ ID NO. 2.

2. A PsIAA27 protein, wherein the PsIAA27 protein comprises a protein expressed by the PsIAA27 gene fragment of claim 1.

3. The PsIAA27 protein according to claim 2, wherein the amino acid sequence of the PsIAA27 protein comprises the amino acid sequence set forth in SEQ ID No. 3.

4. A group of primers for amplifying PsIAA27 gene fragments is characterized by comprising a nucleotide sequence shown in SEQ ID NO.8 and a nucleotide sequence shown in SEQ ID NO. 9.

5. An expression vector comprising the PsIAA27 gene fragment of claim 1.

6. A strain comprising a PsIAA27 gene fragment, comprising the expression vector of claim 5.

7. Use of the PsIAA27 gene fragment of claim 1 or the expression vector of claim 5 for promoting plant rooting.

8. Use according to claim 7, wherein the PsIAA27 gene fragment of claim 1 or the expression vector of claim 5 is used for promoting the growth and/or germination of lateral roots of plants.

9. A method for promoting plant rooting is characterized by comprising the following steps: the PsIAA27 gene was expressed in plants.