CN109609513B - GmLPRF1 for regulating and controlling plant root development and application thereof - Google Patents

Abstract

The invention discloses a gene for promoting the growth of plant root systems, and the nucleotide sequence of the gene is shown as SEQ ID NO: 1. SEQ ID NO: 2 or SEQ ID NO: 3, respectively. It encodes a protein for promoting the growth of plant root systems, and the amino acid sequence of the protein is shown as SEQ ID NO: 4, respectively. According to the inventionGmLPRF1The gene can obviously promote the growth of plant roots. The invention provides a theoretical basis for various plants including soybean participating in a low phosphorus stress response mechanism and the morphological development of roots; meanwhile, a practical model is provided for adapting to low phosphorus stress of plants by changing the root form and configuration; the invention is based on overexpressionGmLPRF1The gene promotes the growth of plant roots, further has important theoretical and practical significance for the adaptation of plants to low phosphorus stress and the revelation of phosphorus on the growth and development of plant roots.

Description

GmLPRF1 for regulating and controlling plant root development and application thereof

Technical Field

The invention relates to the technical field of genetic engineering, in particular to GmLPRF1 regulation and control of plant root development and application thereof.

Background

Terrestrial plants, the biggest difference from animals, are unable to move with environmental changes, and thus plants are important for adaptation to the ever-changing external environment. The root morphology and configuration of terrestrial plants are highly dependent on root configuration, in addition to being responsible for plant anchorage, and also taking into account the uptake of water and nutrients from the soil to supply plant growth. Therefore, the root system shape and configuration of the plant play an important role in the growth of the plant and the adaptation to various adversity stresses.

In order to adapt to various adversity stresses, one strategy is to change the root system shape and configuration, and in the condition of phosphorus deficiency in soil, the arabidopsis thaliana is adapted to low-phosphorus stress, the main root is shortened, and the number of lateral roots and the number of root hairs are increased, so that the contact area with phosphorus is increased, and the phosphorus absorption is improved. In addition, it has been reported that the genotype of phosphorus-efficient kidney beans is largely due to low phosphorus stress to form a large amount of root hairs; under the condition of low phosphorus, the root system, the root surface area and the root volume of the phosphorus-efficient rape variety are greatly increased; the white lupin can generate a large amount of roots under the low-phosphorus condition; in conclusion, phosphorus signals are involved in root morphology and configuration.

Soybean (Glycine max) is an important grain and oil crop in the world, but large areas of acid soil in the world are generally lack of phosphorus, so that the growth and development of the soil are limited and the yield is reduced. As leguminous crops, the fertilizer can be symbiotic with rhizobia to form root nodules, fix nitrogen in air, provide nitrogen sources for plants and improve soil quality. The growth and development of soybean and nitrogen fixation process both need a large amount of phosphorus nutrition. But like other crops, the soybean is also subjected to low-phosphorus stress of soil, so that the growth and development of the soybean are limited, the symbiotic nitrogen fixation is limited, and the yield is reduced. Although the soybean is researched to a certain extent, the soybean is not clear about adapting to low phosphorus stress and changing the root system shape and configuration, and a key gene of the soybean is not found.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and establish a technology and a method for promoting the development of lateral roots of plants and improving the phosphorus absorption capacity of the plants by a transgenic method.

The first purpose of the invention is to provide a gene for promoting the growth of plant root system.

The second purpose of the invention is to provide a protein for promoting the growth of plant root systems.

The third purpose of the invention is to provide the gene, the activator of the gene, the protein and/or the application of the activator of the protein in promoting the growth of plant roots.

The fourth purpose of the invention is to provide the gene, the activator of the gene, the protein and/or the application of the activator of the protein in promoting the plant to adapt to the low phosphorus stress.

A fifth object of the present invention is to provide a method for promoting the growth of a plant root system.

In order to achieve the purpose, the invention is realized by the following technical scheme:

the inventor finds that a RING-FINGER gene which is induced to express by low phosphorus is named as GmLPRF1, an Arabidopsis material which excessively expresses the GmLPRF1 gene is created by utilizing a transgenic technology, and finds that the Arabidopsis material which excessively expresses the GmLPRF1 gene has significantly increased lateral roots and significantly increased lateral root density under normal nutritional conditions compared with Columbia wild type Arabidopsis (Col-0); under low phosphorus conditions, the lateral root number increase is significantly increased and the main root length is significantly increased.

Accordingly, the invention claims the following:

the nucleotide sequence of the gene for promoting the growth of plant roots is shown as SEQ ID NO: 1. SEQ ID NO: 2 or SEQ ID NO: 3, respectively.

SEQ ID NO: the nucleotide sequence shown in 1 is the full length of GmLPRF1 gene, and SEQ ID NO: 2, the nucleotide sequence GmLPRF1 gene transcript shown in SEQ ID NO: 3 nucleotide sequence GmLPRF1 gene CDS.

The protein for promoting the growth of plant roots has an amino acid sequence shown as SEQ ID NO: 4, respectively.

The nucleotide sequence of the genome sequence of GmLPRF1 is shown as SEQ ID NO: 1 is shown in the specification; the nucleotide sequence of the transcript sequence of GmLPRF1 is shown as SEQ ID NO: 2 and the nucleotide sequence of the CDS sequence of the GmLPRF1 is shown as SEQ ID NO: 3, and the amino acid sequence of the GmLPRF1 protein sequence is shown as SEQ ID NO: 4, respectively.

The gene, the activator of the gene, the protein and/or the activator of the protein are/is applied to promoting the growth of plant roots.

Preferably, the plant growth is plant root growth.

Preferably, the root growth is one or more of increased lateral root number, increased lateral root density or main root length.

Preferably, the plant is under normal nutrient conditions and the root system growth is one or both of an increase in lateral root number or an increase in lateral root density.

Preferably, the plant is under low phosphorus conditions and the root growth is one or both of increased lateral root number or major root length.

The application of the gene, the activator of the gene, the protein and/or the activator of the protein in promoting the plant to adapt to low phosphorus stress also belongs to the protection scope of the invention.

A method for promoting the growth of plant root system features that said gene and/or said protein are overexpressed.

The Columbia wild type arabidopsis thaliana is selected as a transgenic receptor material, but the plants comprise various plants including but not limited to arabidopsis thaliana, soybean or other leguminous plants.

Compared with the prior art, the invention has the following beneficial effects:

the GmLPRF1 gene can obviously promote the growth of plant roots. The invention provides a theoretical basis for various plants including soybean participating in a low phosphorus stress response mechanism and the morphological development of roots; meanwhile, a practical model is provided for adapting to low phosphorus stress of plants by changing the root form and configuration; the invention promotes the growth of plant roots by over-expressing GmLPRF1 gene, further has important theoretical and practical significance for the adaptation of plants to low phosphorus stress and the revelation of phosphorus on the growth and development of plant roots.

Drawings

FIG. 1 shows the expression pattern of GmLPRF1 at different phosphorus levels; + P represents normal phosphorus nutrition, 250 μm KH₂PO₄(ii) a P represents a low phosphorus treatment, 12.5 μm KH₂PO₄(ii) a Root denotes RNA extracted from the Root of soybean; leaf denotes RNA extracted from leaves of soybean; data are mean and standard error of 3 biological replicates, asterisks indicate between 2 phosphorus levelsDifference (Student's T-test) indicating significant difference (ρ)<0.05) indicates that the difference is extremely significant (ρ<0.01) indicates that the difference is extremely significant (ρ<0.001)。

FIG. 2 shows the relative expression of GmLPRF1 of transgenic plant material with over-expression GmLPRF 1.

FIG. 3 is the effect of overexpression of GmLPRF1 on root system of Arabidopsis transgenic plants; col-0 is Columbia wild type Arabidopsis thaliana, and GmLPRF1#41, GmLPRF1#45 and GmLPRF1g #78 are overexpression GmLPRF1 strains; a is the number of lateral roots, b is the length of the main root, and c is the density of the lateral roots; + P is normal nutrition condition, -P is phosphorus deficiency treatment, and treatment lasts for 7 days; the data in the figure are mean and standard deviation (SE) of 20 plant samples, asterisks indicate significant differences (ρ <0.05), significant differences (ρ <0.01), and extremely significant differences (ρ <0.001) between different plants and wild type under the same treatment. The experiment was repeated three times with the same trend.

FIG. 4 shows Col-0 being Columbia wild type Arabidopsis thaliana, lines GmLPRF1#41, GmLPRF1#45, GmLPRF1g #78 being overexpression GmLPRF 1; under normal phosphorus nutrient conditions, the treatment lasts for 7 days.

Detailed Description

The invention is described in further detail below with reference to the drawings and specific examples, which are provided for illustration only and are not intended to limit the scope of the invention. The test methods used in the following examples are all conventional methods unless otherwise specified; the materials, reagents and the like used are, unless otherwise specified, commercially available reagents and materials.

Example 1 expression patterns of GmLPRF1 at different phosphorus levels

First, experiment method

(1) Quantitative PCR primer design

Downloading a GmLPRF1 genome sequence (the nucleotide sequence is shown as SEQ ID NO: 1), a GmLPRF1 transcript sequence (the nucleotide sequence is shown as SEQ ID NO: 2), a GmLPRF1CDS sequence (the nucleotide sequence is shown as SEQ ID NO: 3) and a GmLPRF1 protein sequence (the amino acid sequence is shown as SEQ ID NO: 4) from a website;

designing a specific quantitative amplification primer thereof,

GmLPRF1_qF：CAAAGATCTGGGTGAGAAAGG；

GmLPRF1_qR：GAAGTAGCAGGCAAACACTC。

(2) planting and phosphorus deficiency treatment of soybean material

a. Seed disinfection

Mature, filled soybean seeds were arranged in monolayers in petri dishes, placed in a desiccator, and the petri dishes were opened with the lid next to the petri dishes. 100ml of sodium hypochlorite was added to a 250ml beaker in a desiccator, and then 4.2ml of concentrated hydrochloric acid (HCl) was slowly added along the wall of the beaker, and the desiccator was immediately closed and left to stand for 10 hours. Excess chlorine is removed from the dryer.

b. Accelerating germination

And (4) dropping the disinfected seeds into the wet quartz sand for accelerating germination.

c. Transplanting seedlings

After one week, selecting normal seedlings with consistent growth vigor from two leaves and one heart, transplanting the seedlings to a water culture system, and culturing the seedlings in a greenhouse by using 1/2 soybean nutrient solution.

d. Treatment of

After the first three compound leaves are completely unfolded for about one week, the treatment is carried out: low phosphorus (LP: 25. mu.M), high phosphorus (HP: 500. mu.M), 5 biological replicates per treatment. The nutrient solution was changed once a week and the pH was adjusted once every 3 days.

e. Sampling

The roots and leaves 14 days after treatment were sampled, frozen in liquid nitrogen and stored at-80 ℃.

(3) Extraction of Total RNA

Total RNA was isolated in one step with reference to TRIzo 1. Putting 0.2g of sample into a precooled mortar, grinding into powder, transferring into a 1.5ml centrifuge tube, adding 1ml of TRIzo1 extracting solution, violently shaking and uniformly mixing, and standing at room temperature for 5 minutes; adding 0.2ml of chloroform, shaking vigorously, standing at room temperature for 2-5 minutes, and centrifuging at 12000rpm at 4 ℃ for 15 minutes; transferring the supernatant into a new tube, adding 0.5ml of isopropanol, standing at room temperature for 10 minutes, centrifuging at 4 ℃ at 12000rpm for 5 minutes, pouring off ethanol, air-drying the precipitate, and adding a DEPC aqueous solution; finally, the OD value is measured to determine the purity and concentration of the RNA.

(4) Preparation and analysis of real-time fluorescent quantitative PCR sample

Total RNA was treated with dnase i to remove contamination of genomic DNA and RNA was inverted to the first strand as per the reverse transcriptase instructions. The resulting first strand was diluted 100-fold and used as a template for quantitative PCR reactions. And (3) performing gradient dilution on a proper amount of cDNA stock solution to form a template of a standard curve. The reaction system and the reaction conditions are shown in Table 1 and Table 2, respectively.

Table 1 quantitative PCR reaction system (total volume 20. mu.L):

table 2 quantitative PCR reaction conditions:

the expression level of each sample was calculated using Real-time analysis Software 6.0 from Rotor-Gene.

Second, experimental results

The soybean housekeeping gene GmEF1a was used as a reference gene. The relative expression level is the ratio of the expression level of the target gene to the expression level of the housekeeping gene. Expression patterns of GmLPRF1 at different phosphorus levels (see fig. 1), and fig. 1 shows that the GmLPRF1 gene was induced to express under low phosphorus treatment, whether in the soybean leaf or root.

Example 2 construction of GmLPRF1 Gene overexpression vector (gateway method)

First, experiment method

(1) Primer design

Downloading CDS sequence (nucleotide sequence is shown as SEQ ID NO: 3) of GmLPRF1 gene from a website, designing specific amplification primers thereof,

OE-GmLPRF1F：ttgtacaaaaaagcaggctggATGACTTCCTCACTCGTGAA；

OE-GmLPRF1R：gtacaagaaagctgggtaTCACTTCCTACCCAACCTTAT。

(2) fragment amplification and purification

The cDNA of soybean YCO3-3 is taken as a template, a corresponding fragment is amplified by using a specific primer of OE-GmLPRF1, and a reaction system and reaction conditions are shown in the following tables 3 and 4; and running the PCR reaction product on agarose gel electrophoresis, cutting and recovering a bright band with a consistent position, and recovering the PCR product by using an agarose gel DNA recovery kit to obtain a target fragment.

Table 3 PCR reaction system (total volume 50 μ L):

table 4 PCR reaction conditions:

(3) ligation transformation of the intermediate vector pDONR207

Connecting the recovered target fragment with a pDONR207 vector, transforming Escherichia coli DH10B, selecting a single clone, carrying out PCR detection on a pipeshake, sending the obtained product to a sequencing company for sequencing, and connecting an intermediate vector with a system shown in the following table 5;

table 5 intermediate loaded ligation system (total volume 10 μ L):

(4) target vector pMDC32 ligation transformation

And (3) correctly sequencing, extracting the plasmid by using a small-amount plasmid rapid extraction kit, connecting the plasmid with a target fragment with a target vector pMDC32, transforming the plasmid into Escherichia coli DH10B, selecting a single clone, performing PCR (polymerase chain reaction) detection on the pipeshake, and sequencing by a sequencing company. And (4) after the sequencing is correct, respectively storing the bacterial liquid and the plasmid for later use. Meanwhile, the recombinant target vector is transformed into an agrobacterium strain GV3101, and a bacterial solution is stored for later use after detection without errors, wherein a target vector connection system is shown in the following table 6.

Ligation system for loading of table 6 (total volume 10 μ L):

second, experimental results

The Agrobacterium strain GV3101 carrying the recombinant vector of interest was successfully constructed.

Example 3 Arabidopsis thaliana genetic transformation

First, experiment method

(1) Planting of wild type Arabidopsis thaliana (Col-0)

Uniformly sowing wild type Arabidopsis seeds (Col-0) on the surface of a nutrition pot filled with a substrate, spraying a certain amount of water, covering with a plastic film for moisturizing, and placing in the dark at 4 ℃ for 2-3 days to break seed dormancy.

Then placing the seedlings in an artificial climate chamber (16 hours of illumination 24 ℃/8 hours of darkness 24 ℃) for culture, after the length is equal for one week, respectively transplanting the seedlings into nutrition pots, wherein 3-4 seedlings are planted in each pot, when 1cm of the first bolting is carried out, the main stems are cut off to increase the branches, and when the next arabidopsis is full of flowers, the seedlings can be transformed by using agrobacterium liquid.

(2) Transformation of Arabidopsis thaliana floral battings with Agrobacterium solution

Carrying out scribing activation on the agrobacterium GV3101 preservation solution carrying the target vector, firstly inoculating the agrobacterium GV3101 preservation solution to a 50ml YEP (Kan + Rif) culture plate, and culturing for 2 days at the temperature of 28 ℃; after the growth of the bacteria, single clone was picked up, the bacteria were shaken in a small tube for 2ml of YEP (Kan + Rif) liquid, and then the Agrobacterium liquid was subjected to amplification culture in 500ml of YEP (Kan + Rif) liquid at 28 ℃ for 20 hours at 200 rpm. The broth was centrifuged at 4000rpm for 10 minutes, the supernatant was discarded, and the Agrobacterium was resuspended in one volume of fresh impregnation solution and then transferred to a 500ml beaker. Immersing the overground part of the arabidopsis containing flowers in the agrobacterium liquid for 10 seconds, slightly stirring, taking out the soaked plants, covering the transformed arabidopsis with a plastic cover, laterally placing for 16-24 hours to ensure higher humidity, removing the plastic cover the next day, placing in an arabidopsis room for growing till the seeds are mature, and collecting T0 generation seeds; the immersion fluid formulation is shown in Table 7 below;

table 7 immersion fluid formulations:

(3) transgenic Arabidopsis progeny identification

Sterilizing mature T0 generation seeds, sterilizing with 75% alcohol, sterilizing with 100% alcohol, and blow-drying; sowing the disinfected seeds on an 1/2MS culture medium (containing hygromycin x1000), and screening transgenic arabidopsis thaliana of T1 generation, wherein the main root of the arabidopsis thaliana seedling is long, which indicates that the hygromycin is resistant and possibly is a transgenic material; and then, the arabidopsis seedlings with the main roots are transferred to a substrate to grow, T1 generation transgenic material seeds are harvested, the previous step is repeated, hygromycin is used for screening, whether the length of the main roots of the arabidopsis seedlings meets 3:1 (single copy insertion) or not is judged, if yes, the arabidopsis seedlings can be determined to be transgenic materials, and T2 generation arabidopsis seeds are harvested after the arabidopsis seedlings are mature.

During the growth period of T2 generation transgenic Arabidopsis, over-expression transgenic lines are sampled to extract RNA, and after reverse transcription into cDNA, the relative expression amount of genes is detected by fluorescence quantitative PCR. The method comprises the following specific steps: leaf picking of Arabidopsis rosette leaves grown on a substrate for 14 days is carried out by 0.1g, frozen with liquid nitrogen and stored at-80 ℃. Then extracting RNA by using a kit for extracting RNA, carrying out reverse transcription to obtain cDNA, and finally detecting the relative expression quantity of the gene by using fluorescent quantitative PCR.

Second, experimental results

FIG. 2 shows the expression level detection of transgenic plant material GmLPEF1 of over-expressed GmLPRF 1. The results show that: the expression level of GmLPRF1 in the strain overexpressing GmLPRF1#41, #45, #78 is obviously higher than that of the wild type, wherein the strains of OE-GmLPRF1#41, OE-GmLPRF1#45 and OE-GmLPRF1#78 are 292 times, 4435 times and 461 times of that of the wild type respectively, and statistical analysis shows that the difference reaches a significant level (rho <0.05) or a very significant level (rho < 0.01).

Example 3 GmLPRF1 functional analysis experiment

First, experiment method

(1) Seed disinfection

Putting mature and full arabidopsis seeds into a 2ml tube, sucking 1ml of 75% alcohol into the 2ml tube on a super clean bench, covering the seeds, uniformly stirring, sucking out the alcohol, repeating the previous step for no more than 1 minute, then sterilizing the seeds for the second time by using 100% alcohol, and then putting the seeds on the super clean bench for drying.

(2) Seed germination

The sterilized seeds were sown on a normally nutritious 1/2MS plate, then placed in dark at 4 ℃ for 2-3 days to break seed dormancy, and then placed in a climatic chamber (16 hours light 24 ℃/8 hours dark 24 ℃) for cultivation.

(3) Transplanting seedlings and treating with phosphorus nutrition

Three days after seed germination, 20 consistent growing Arabidopsis seedlings were individually transferred to high phosphorus, low phosphorus treated 1/2MS plates and then placed in a climatic chamber (24 ℃ C. for 16 hours under light/24 ℃ C. for 8 hours in dark 24 ℃ C.) for culture. High phosphorus (P: 0.625mM), low phosphorus (P: 0mM), each treatment of 20 samples, three replicates. The growth is carried out for 7 days. Measurement indexes are as follows: lateral root number, main root length, lateral root density.

(4) Influence of overexpression of GmLPRF1 on root systems of transgenic Arabidopsis plants.

And after the arabidopsis grows for 7 days, detecting the lateral root number, the main root length and the lateral root density of the underground part of the plant.

Second, experimental results

FIGS. 3 and 4 are graphs showing the effect of overexpression of GmLPRF1 on root system and root number of transgenic Arabidopsis plants. The results show that: under normal nutritional conditions, the lateral root number of three strains (#41, #45, #78) overexpressing GmLPRF1 is greatly increased compared with the wild type; the length of the main root of the strain of the GmLPRF1 is not significantly different from that of the wild type; the lateral root density of the strain overexpressing GmLPRF1 was significantly increased compared to the wild type.

Under low phosphorus conditions, the number of lateral roots of three strains (#41, #45, #78) overexpressing GmLPRF1 was significantly increased compared to the wild type; meanwhile, compared with the wild type, the length of the main root of the strain of the GmLPRF1 is increased, wherein the length of the main root is significantly increased compared with the wild type in the strains #41 and # 78; however, the lateral root density of the strain overexpressing GmLPRF1 was not significantly different compared to the wild type.

Sequence listing

<110> southern China university of agriculture

<120> GmLPRF1 regulation and control of plant root development and application thereof

<160> 4

<170> SIPOSequenceListing 1.0

<210> 1

<211> 5300

<212> DNA

<213> Glycine max

<400> 1

cttttcctgt gtgtctggtg gtgagtttaa aagtttggaa ttgacattgt ctttgtttta 60

gtgttttcct tgatcccttt ggcataacag cattattctt cagcatctct cgtattcctt 120

ctccttgtgt gtctctcctt gctttatctc tcttaatttc ttcagatttt agttttctat 180

tgtatggttc tgcgttttcg cacaactcac tatagtttct tctatacctt cctggtggat 240

ctgtcactat gacttcctca ctcgtgaaga tccaaggtga acaggttctc accaacgact 300

ttcaggacct ttcaatcaaa gatctggtat tttccccttc cttcttgtta atgggtatgg 360

atatttcttc cttctttgta atgggtatgc cttgttttga ttaaattcat attttgaatt 420

tctggtttgt ttggacgatt ttgggggtac ccaattgggt tttcatgtga aagtttcaaa 480

atttatgttg tgggcatgat ttgtttttaa ccaattgttg attttgttga tttgtttttt 540

gggttggaaa tgttagaatt ggtgttcctt ttgttgatgc catggttttt tttatgctgt 600

ttttgttcag ggtgagaaag gaagtgaggc agagattcac gaggtggggt atggaggcca 660

tgggggaatc tgtgctattt gcttggataa gatagtgctg caggaaactg ctcttgtaaa 720

aggttgcgag catgcttact ggtttgtctg aacgatctct tgctgttgtt ttcagaatta 780

agtttgggga aaatgtgcct ttttatttat ttattattat ggtatgttta ttttacgatt 840

gctttaggat tggttggaat tttcagtact aattgttgtt ctcagggata tggattagtg 900

ccactttgca aatgtggtgg tatcttttct tccttgaacc aaatgaaaaa aaaacctcca 960

tctcttagta tgttttctgt gtttatcctg atgatgtggg aatgattcat ttaattggag 1020

atgaattatg tctgagctag aaagatcatt tatgtttgga tttttatttt cttcatatct 1080

tttgtgcttg agatttgaga tcttttaaag tcaagggtaa acttgtggct ttactacctt 1140

ttgtttgtac tttgatttgc aaacatctat ttatgagttt gttgtaagtt tgtcacatcg 1200

aaaaagacaa agcattatat aggttcagtt agtacgttaa ttgagcatta tggattccca 1260

caaggtagat tgagaggttc tgatgactga tgcggtcatc tgtaatccta gttcagttag 1320

tatgtaaacc gttggatctc aaacatattc ctgtatgtac tatcaaggtt atcaatggaa 1380

gatggtggaa ggccaaaatt ctgccatatg agcctgccat ggcagcctat ggtgctgcca 1440

tagcgggcct cccttcacaa attgcctata gcggaagggt tgaaaaatgc cacattatag 1500

cattttggtg atgctataat ggttaattga caacattggt acttcttttt ttatggttgt 1560

caaggttgaa gtctccatga tttggatggt tagtggcaga tagtaactcc cttaaaagct 1620

caagatgcaa gagcactttg ggcttgaagg ctggagaatg tacaagtgtt attttaacat 1680

gttccgagcc tcaactttta tttagaagaa tggatgagat agattgaatt tgaactcacg 1740

gcctcttggt agtagagatt ctgatattat ctatgttata aaagtatatg ttgttaggtg 1800

aaactgtgaa agcacatgaa tggttttatt taatgtttaa tgactaggtt agctttgatt 1860

tgatagttga tagtgtttcg aatttacacc caatatttgt tcaatactga aatgataata 1920

tatctaggtt ttaccccaac atgtatccca tattgaaagc acacttacag atgtcattat 1980

atttacaaat gatagctggt ttggcacttt aggtagtaaa gtttcttgca gttgcaatga 2040

gttggaaatt agaattgatt cttacttttc cttggagaat gggatgttgc ttcacacttt 2100

ttcagctgtt taatctacat tcatggatga gaggagttct tcataattta cattcaatca 2160

tcatatcttg tctcttcaag ttatcttagt tgatcaggga tgatatacat agcatgttca 2220

attgaatcta tggagttgag tgatcaacag tgctttatat ggctgaggca aatcgtttag 2280

gatcttatat ttcttgtgaa tgaatatcct agaatgtgtc tgcatctgat gtgatttgtg 2340

ttggttctgc atcagtgaga ttgaaaaaat ataggtccaa aacctggatt gataagctaa 2400

ctgactttta gtggacatat tttagttaat ttttaatgct cttgaccctt ctttgtatgt 2460

tggggaagtt ccctcactta aatttcagca taaaacagga ggtttttttg catttttact 2520

tttctgttct taatgtatat ttccttttgg ctaattgcag gtacacactg tagacaattt 2580

tgttatattt tgattttgta gtattttcca acttttgttc catgcattat ctttcacggt 2640

aatggaaact acttgtgtag ttggaaattc tagtcattga ttactaacat tccatgcatt 2700

atctttctat tttttttttt tttttaaatt cactactatt gaattgactg ctttggcttg 2760

agattattat tattattatt tttgcgtttt ctacatcttt ttgtttatgg ttttaattga 2820

ttttgtgtag tgtaacatgc atccttcatt gggctacata ccgtgagaaa gttacctgcc 2880

ctcagtgtaa acatccattt gagttcctca atgtccatcg ctcacttgat ggcaggtaat 2940

aattgaacct tttgcatcgg ttttcatttc taatttagtt cttgctactt atgtttaaaa 3000

cgtgatggaa agaagtgatt atagttttgt gtgagtggat cctctcattt taaatttcac 3060

atgagagaga gagagagagt acatgatgag gtcatgaaat ttcaacatca gaggattcgt 3120

tcagattttt tgtatgtgat cagtttagca tgcttcgctg taatcaacaa tgaaaggatg 3180

taaaagatat attataattt gagattttct tttgaaaata cttgagttat tatggaaaat 3240

gagacttcta attttatctg gttgaagtta ggttactgtg ttatcatgtt tacctgaccc 3300

ctaaacaata tgttgggagg ctgctggact ttctagtttt caatttaata caagttagaa 3360

cctaaaaaaa ttggtttgga ctccaaaaat agaatgatat tctggtttag tgcttgttag 3420

ggggtttccc ccattttttc catcaaaaaa atattagggg atgttcggtt tgattgtttt 3480

ctgttttcat tttcattgaa aacagaaaac ggtgatgaaa atgtgtttgg ttggatttct 3540

gaaaacattt tcagtgaaaa tgaaaacagg aaacaaccaa aaaatgaaaa taataaattc 3600

tcattttcag tgttttcaat tgagaacaaa aatctcattt tgggtaaaat gaaattgcgg 3660

tgacaatgaa tgtaatttta agcaaatata aaaatacaaa aagacaagaa atcaatatat 3720

catagatttt cagtattttt atttcatgaa aatagaaaat aagaagtcaa actaaacatg 3780

ttttcagaat tctaatattt tgaaaatgaa aatagttttc agaaaatgaa aacagaaaat 3840

gagaacagaa aatgaaaatg caaacaaaac acatccttag caatcttatt ttaaacattg 3900

acaattttac ttttgtcaac ctttttagct cctttggtta tataccataa aatactatca 3960

tctcttagta cagaaacttt aagggggtgt ttggtttggt tgttttctgt tttcattttc 4020

actcgaaata gaaaatgatg atgaaaatgt gtttggttga atttctgttt tcagtttcag 4080

tgaaaatatt tttcctaacg aaccaaaaac tggaaacaat aaaatctcgt tttcagttga 4140

aaccaagaac ctcattttag gtaaaatgaa aacgcggtgg caaagaatgt aattttaagc 4200

aaatctaaaa atacatttcc ttttgaaaac gcgttttcag tgtttttatt tcttgaaagc 4260

ataaaacaag aagtcaaacc aaacatgttt ttaaaatttt aatcttttga aaatgaaaat 4320

agttttcaga aaatgaaaac aggaaatgaa aatgcaaatg caaaccaaac acaccctaaa 4380

atacttaaga ctagggtcta acaaacaaaa tgccaaaata atgtagcatc ggcaactact 4440

taaatttttg tacctataat tgcagaatga acattctata ggtgaagtgt tgcattagaa 4500

tgtctgtttg gcaatcactc caagttgtgt ctgaaatcat gttcttatta cttgatcttc 4560

aattatactg tggcagcatt caagattaca tgtttgagga gagtgtttgc ctgctacttc 4620

gagcctcatg gttcacacct ttatctgttg aagaacatgt ggctcatgaa gatgtatatg 4680

aagacctaga agattattat cagtacgagg atgatgatga tatggatgaa gcttactatg 4740

gtggttcgtc caatcttcgc gttattggca accgtagatg gggagataat ggttatgtca 4800

gggctggacg tcaagaagct cggccagtcc atcgtctgaa cttccaggat tcaggagcaa 4860

gttcttcttc gcgcgagccg aagaagaaag aggttgggaa aattatcaca ggcagaaggg 4920

caaagagggc acagaaaagg gaagcggctg acaaggcggc cgaagcaaag catctgcagc 4980

atttaataag gttgggtagg aagtgattct attttattta tatcaggcca tcatcatggc 5040

cttgaacttt ccttcccata agttaaatat tttctttttt ttatatattt tttttttctt 5100

gtacattgtt tctgtgctct ttactctgaa gcatgtaata tatcttgggg aagttgtttt 5160

agaataatgg cttggtgggg tggttggatt atggaaattc caaactcact ataccatgaa 5220

caatttaggt tatatttagt tattatgctc aatgcaggtc taagttgtac ataatttcct 5280

tgtgatgttc ttgtgatgtt 5300

<210> 2

<211> 1350

<212> DNA

<213> Glycine max

<400> 2

cttttcctgt gtgtctggtg gtgagtttaa aagtttggaa ttgacattgt ctttgtttta 60

gtgttttcct tgatcccttt ggcataacag cattattctt cagcatctct cgtattcctt 120

ctccttgtgt gtctctcctt gctttatctc tcttaatttc ttcagatttt agttttctat 180

tgtatggttc tgcgttttcg cacaactcac tatagtttct tctatacctt cctggtggat 240

ctgtcactat gacttcctca ctcgtgaaga tccaaggtga acaggttctc accaacgact 300

ttcaggacct ttcaatcaaa gatctgggtg agaaaggaag tgaggcagag attcacgagg 360

tggggtatgg aggccatggg ggaatctgtg ctatttgctt ggataagata gtgctgcagg 420

aaactgctct tgtaaaaggt tgcgagcatg cttactgtgt aacatgcatc cttcattggg 480

ctacataccg tgagaaagtt acctgccctc agtgtaaaca tccatttgag ttcctcaatg 540

tccatcgctc acttgatggc agcattcaag attacatgtt tgaggagagt gtttgcctgc 600

tacttcgagc ctcatggttc acacctttat ctgttgaaga acatgtggct catgaagatg 660

tatatgaaga cctagaagat tattatcagt acgaggatga tgatgatatg gatgaagctt 720

actatggtgg ttcgtccaat cttcgcgtta ttggcaaccg tagatgggga gataatggtt 780

atgtcagggc tggacgtcaa gaagctcggc cagtccatcg tctgaacttc caggattcag 840

gagcaagttc ttcttcgcgc gagccgaaga agaaagaggt tgggaaaatt atcacaggca 900

gaagggcaaa gagggcacag aaaagggaag cggctgacaa ggcggccgaa gcaaagcatc 960

tgcagcattt aataaggttg ggtaggaagt gattctattt tatttatatc aggccatcat 1020

catggccttg aactttcctt cccataagtt aaatattttc ttttttttat atattttttt 1080

tttcttgtac attgtttctg tgctctttac tctgaagcat gtaatatatc ttggggaagt 1140

tgttttagaa taatggcttg gtggggtggt tggattatgg aaattccaaa ctcactatac 1200

catgaacaat ttaggttata tttagttatt atgctcaatg caggtctaag ttgtacataa 1260

tttccttgtg atgttcttgt gatgttgata tcattgacac ctcaacaatg tctatatcta 1320

tctctttctt caaagttaag aatttcatac 1350

<210> 3

<211> 744

<212> DNA

<213> Glycine max

<400> 3

atgacttcct cactcgtgaa gatccaaggt gaacaggttc tcaccaacga ctttcaggac 60

ctttcaatca aagatctggg tgagaaagga agtgaggcag agattcacga ggtggggtat 120

ggaggccatg ggggaatctg tgctatttgc ttggataaga tagtgctgca ggaaactgct 180

cttgtaaaag gttgcgagca tgcttactgt gtaacatgca tccttcattg ggctacatac 240

cgtgagaaag ttacctgccc tcagtgtaaa catccatttg agttcctcaa tgtccatcgc 300

tcacttgatg gcagcattca agattacatg tttgaggaga gtgtttgcct gctacttcga 360

gcctcatggt tcacaccttt atctgttgaa gaacatgtgg ctcatgaaga tgtatatgaa 420

gacctagaag attattatca gtacgaggat gatgatgata tggatgaagc ttactatggt 480

ggttcgtcca atcttcgcgt tattggcaac cgtagatggg gagataatgg ttatgtcagg 540

gctggacgtc aagaagctcg gccagtccat cgtctgaact tccaggattc aggagcaagt 600

tcttcttcgc gcgagccgaa gaagaaagag gttgggaaaa ttatcacagg cagaagggca 660

aagagggcac agaaaaggga agcggctgac aaggcggccg aagcaaagca tctgcagcat 720

ttaataaggt tgggtaggaa gtga 744

<210> 4

<211> 247

<212> PRT

<213> Glycine max

<400> 4

Met Thr Ser Ser Leu Val Lys Ile Gln Gly Glu Gln Val Leu Thr Asn

1 5 10 15

Asp Phe Gln Asp Leu Ser Ile Lys Asp Leu Gly Glu Lys Gly Ser Glu

20 25 30

Ala Glu Ile His Glu Val Gly Tyr Gly Gly His Gly Gly Ile Cys Ala

35 40 45

Ile Cys Leu Asp Lys Ile Val Leu Gln Glu Thr Ala Leu Val Lys Gly

50 55 60

Cys Glu His Ala Tyr Cys Val Thr Cys Ile Leu His Trp Ala Thr Tyr

65 70 75 80

Arg Glu Lys Val Thr Cys Pro Gln Cys Lys His Pro Phe Glu Phe Leu

85 90 95

Asn Val His Arg Ser Leu Asp Gly Ser Ile Gln Asp Tyr Met Phe Glu

100 105 110

Glu Ser Val Cys Leu Leu Leu Arg Ala Ser Trp Phe Thr Pro Leu Ser

115 120 125

Val Glu Glu His Val Ala His Glu Asp Val Tyr Glu Asp Leu Glu Asp

130 135 140

Tyr Tyr Gln Tyr Glu Asp Asp Asp Asp Met Asp Glu Ala Tyr Tyr Gly

145 150 155 160

Gly Ser Ser Asn Leu Arg Val Ile Gly Asn Arg Arg Trp Gly Asp Asn

165 170 175

Gly Tyr Val Arg Ala Gly Arg Gln Glu Ala Arg Pro Val His Arg Leu

180 185 190

Asn Phe Gln Asp Ser Gly Ala Ser Ser Ser Ser Arg Glu Pro Lys Lys

195 200 205

Lys Glu Val Gly Lys Ile Ile Thr Gly Arg Arg Ala Lys Arg Ala Gln

210 215 220

Lys Arg Glu Ala Ala Asp Lys Ala Ala Glu Ala Lys His Leu Gln His

225 230 235 240

Leu Ile Arg Leu Gly Arg Lys

245

Claims (5)

1. The nucleotide sequence is shown as SEQ ID NO: 1 or SEQ ID NO: 2 in promoting the growth of plant root systems.

2. The use of claim 1, wherein the root growth is one or more of increased lateral root count, increased lateral root density, or increased main root length.

3. The use of claim 2, wherein the plant is under normal nutrient conditions and the root growth is one or both of an increase in lateral root number or an increase in lateral root density.

4. The use of claim 2, wherein the plant is under low phosphorus conditions and the root growth is one or both of increased lateral root count or major root length.

5. A method for promoting the growth of plant roots, characterized in that the overexpression nucleotide sequence is as shown in SEQ ID NO: 1 or SEQ ID NO: 2, or a pharmaceutically acceptable salt thereof.

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