CN109608530B - Soybean low-phosphorus response gene for promoting lateral root formation, protein and application thereof - Google Patents

Abstract

The invention discloses a soybean low-phosphorus response gene for promoting lateral root formationNFYA7And the protein and the application thereof, wherein the amino acid sequence of the GmNFYA7 protein is shown as SEQ ID NO: as shown in (4) in the figure,GmNFYA7the nucleotide sequence of the gene is shown as SEQ ID NO: 1. SEQ ID NO: 2 or SEQ ID NO: 3, respectively. The invention provides a theoretical basis for leguminous crops including soybean and dicotyledonous plants on the basis of the mechanism that NF-YA regulatory factors participate in the plant response to low phosphorus stress and change the configuration and the form of a root system. Meanwhile, the invention provides an effective method for plants to adapt to low phosphorus stress by changing the configuration and the shape of the root system, has great commercial value for crops, and is worthy of popularization and application.

Description

Soybean low-phosphorus response gene for promoting lateral root formation, protein and application thereof

Technical Field

The invention relates to the technical field of genetic engineering, in particular to a soybean low-phosphorus response gene for promoting lateral root formation, and a protein and application thereof.

Background

The root system is the key for the terrestrial plants to fix in soil and effectively absorb water and mineral nutrients, and plays an important role in the aspect of adapting to various adversity stresses. Root System Architecture (RSA) is the overall spatial layout of the various parts of the root system and is an important factor in determining the availability of resources to plants. Plant productivity is greatly affected by the structural characteristics of the root system, which are important targets for agricultural improvement. The change of the structural characteristics of the root system can change the contact of the plant root system to different layers in heterogeneous soil, so that the plant can effectively absorb more nutrients and water. This is because resources like water and nitrogen are usually found in deeper soil layers, while other nutrients like phosphorus are more abundant in shallower soil layers. Therefore, the configuration and the shape of the plant root system have very important functions on the growth and the development of the plant and the adaptation to various adversity stresses.

Current research shows that: phosphorus signals are involved in altering plant root architecture and morphology. Plants respond in a series to maintain self-phosphorus homeostasis under phosphorus deficiency conditions, and one important response is root architecture remodeling.

Research shows that in a low-phosphorus environment, the root system can change the morphological characteristics of the root system, improve the absorption capacity of insoluble phosphorus in soil, help plants to better absorb and utilize nutrients in the low-phosphorus environment and enhance the adaptability of the plants in adverse circumstances. For example, the low phosphorus stress causes the soybean root length to be longer and the root surface area to be increased; under the stress of phosphorus, the cowpea can enlarge the absorption range of the root system by increasing the length of the root and the number of lateral roots; under the stress of low phosphorus, the growth of the main root of the arabidopsis thaliana is inhibited, and the number of lateral roots and the number of root hairs are increased, so that the contact area of the plant and the phosphorus is increased, and the absorption of the phosphorus is improved; alfalfa can cope with low phosphorus stress by increasing the number and length of root hairs, and can also obtain more phosphorus from soil for plant growth by establishing mycorrhizal tissues or forming row roots; under the condition of phosphorus deficiency, the growth of stems of corn and broad bean is obviously inhibited, and the root growth and the total length of roots are increased. In conclusion, phosphorus stress can be involved in the alteration of plant root system configuration and morphology.

Soybean (Glycine max) is an important protein and oil resource for human beings and animals, and is also a high-phosphorus-demand variety, so that the sensitivity to low phosphorus stress is a main limiting factor for soybean production. The soybean, as a leguminous crop, can symbiotic with rhizobia to form nodules to perform biological nitrogen fixation reaction, and the reaction has the effects of losing weight, increasing yield, fertilizing soil, improving soil quality and the like. Low phosphorus stress has many effects on soybean growth, such as the growth of soybean root nodules and nitrogen fixation are reduced, flower pod shedding is increased, and the overall growth and development of plants are affected, so that the yield and the seed quality are affected.

NF-YA is a transcription factor existing in eukaryote, and in recent years, it is found that NF-YA participates in a plurality of physiological processes such as plant embryonic development, photosynthesis, flowering time, adversity stress response and the like, and plays an important role in regulation. At present, soybeans have been studied to a certain extent, but the mechanism for adapting to low-phosphorus stress and changing root system configuration and form and the mechanism for NF-YA transcription regulation factors to participate in related regulation processes are not clear yet.

Disclosure of Invention

The invention aims to overcome the defects that the prior art is not clear about the mechanism that soybeans adapt to low-phosphorus stress and change the configuration and the form of a root system and NF-YA transcription regulation factors participate in related regulation processes, and provides a soybean low-phosphorus response gene for promoting the formation of lateral roots, and protein and application thereof.

The invention aims to provide application of GmNFYA7 protein or an activator thereof in promoting plant root growth.

The second purpose of the invention is to provide the application of the GmNFYA7 gene or the activator thereof in promoting the growth of plant roots.

The third purpose of the invention is to provide the application of the GmNFYA7 protein or the activator thereof in promoting the resistance to low phosphorus stress.

The fourth purpose of the invention is to provide the application of the GmNFYA7 gene or the activator thereof in promoting the low phosphorus stress resistance.

It is a fifth object of the present invention to provide a method for promoting plant root growth, increasing the number of lateral roots in a plant, and/or promoting the break through of lateral root primordia through the epidermis.

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

the inventor finds that an NF-YA gene expressed by low phosphorus induction is named GmNFYA7, and supposes that the NF-YA gene is involved in soybean low phosphorus signal transduction. Researches show that compared with Columbia wild type arabidopsis (Col-0), the arabidopsis material with the overexpression GmNFYA7 gene can obviously promote lateral root primordium to break through epidermis, improve the number of lateral roots and possibly promote phosphorus absorption under the condition of low phosphorus by utilizing the arabidopsis material with the overexpression GmNFYA7 gene by utilizing a transgenic technology.

The invention therefore claims the following:

the application of the GmNFYA7 protein or an activator thereof in promoting the growth of plant roots is characterized in that the amino acid sequence of the GmNFYA7 protein is shown as SEQ ID NO: 4, respectively.

The application of the GmNFYA7 gene or an activator thereof in promoting the growth of plant roots is characterized in that the nucleotide sequence of the GmNFYA7 gene is shown as SEQ ID NO: 1. SEQ ID NO: 2 or SEQ ID NO: 3, respectively.

Wherein the nucleotide sequence of the GmNF-YA7 genome sequence is shown as SEQ ID NO: 1 and the nucleotide sequence of the GmNFYA7 transcript sequence is shown as SEQ ID NO: 2 and the nucleotide sequence of the CDS sequence of GmNFYA7 is shown as SEQ ID NO: 3, and the amino acid sequence of the GmNF-YA7 protein sequence is shown as SEQ ID NO: 4, respectively.

Preferably, the promoting the growth of the plant root system is increasing the number of lateral roots.

Preferably, the increase in lateral root number is due to the breakthrough of lateral root primordia through the epidermis.

Preferably, the plant is a dicot.

The application of GmNFYA7 protein or an activator thereof in promoting low-phosphorus stress resistance; the application of the GmNFYA7 gene or an activator thereof in promoting the low-phosphorus stress resistance also belongs to the protection scope of the invention.

The invention also claims a method for promoting the growth of plant roots, increasing the number of lateral roots of plants and/or promoting lateral root primordia to break through epidermis, wherein GmNFYA7 gene and/or GmNFYA7 protein are overexpressed in the plants.

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 invention provides a theoretical basis for leguminous crops including soybean and dicotyledonous plants on the basis of the mechanism that NF-YA regulatory factors participate in the plant response to low phosphorus stress and change the configuration and the form of a root system. Meanwhile, the invention provides an effective method for plants to adapt to low phosphorus stress by changing the configuration and the shape of the root system, has great commercial value for crops, and is worthy of popularization and application.

Drawings

FIG. 1 shows the expression pattern of GmNFYA7 gene 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 in the graph are mean and standard error of 3 biological replicates, asterisks indicate 2 differences between phosphorus levels (Student's T-test), indicating significant differences (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 level of GmNFYA7 gene of transgenic Arabidopsis with GmNFYA7 gene overexpressed;

FIG. 3 shows the effect of overexpression of GmNFYA7 gene on root system of Arabidopsis transgenic plant; col-0 is Columbia wild type arabidopsis, OE-GmNFYA7#5 and OE-GmNFYA7#20 are overexpression GmNFYA7 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; HP is normal nutrition condition, LP is phosphorus deficiency treatment, and the treatment lasts for 7 days; the data in the figure are the mean and standard deviation (SE) of 12 samples, with asterisks indicating differences between different plants and wild type under the same treatment (Student' st-test); significant differences (, 0.05),.; very significant differences (, 0.01),; extremely significant differences (, 0.001).

FIG. 4 is a graph of the number of lateral roots of WT and OE-GmNFYA7#5, OE-GmNFYA7#20 on low phosphorus treatment; col-0 is Columbia wild type arabidopsis, OE-GmNFYA7#5 and OE-GmNFYA7#20 are overexpression GmNFYA7 strains; the data in the figure are the mean and standard deviation (SE) of 12 biological replicates, 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 (Student' st-test)

FIG. 5 is a diagram showing the effect of overexpression of GmNFYA7 on the lateral root number of transgenic Arabidopsis under low-phosphorus conditions; col-0 is Columbia wild type arabidopsis, OE-GmNFYA7#5 and OE-GmNFYA7#20 are overexpression GmNF-YA7 strains; and treating for 7 days under low-phosphorus conditions.

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 GmNFYA7 Gene quantification experiment

First, experiment method

1. Quantitative PCR primer design

The genome sequence of GmNFYA7 (the nucleotide sequence is shown as SEQ ID NO: 1), the transcript sequence of GmNFYA7 (the nucleotide sequence is shown as SEQ ID NO: 2), the CDS sequence of GmNFYA7 (the nucleotide sequence is shown as SEQ ID NO: 3) and the protein sequence of GmNF-YA7 (the amino acid sequence is shown as SEQ ID NO: 4) are downloaded from a website.

Designing a specific quantitative amplification primer:

GmNFYA7.qF：CATGACCTCTCAGATAATGAAGC；

GmNFYA7.qR：TGCAAGTATGGCTTCCGAT。

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. Removing excess chlorine gas after removal from the dryer;

b. accelerating germination

The disinfected seeds are dropped into wet quartz sand for accelerating germination;

c. transplanting seedlings

After one week, selecting normal seedlings with consistent growth vigor, 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:

table 2 quantitative PCR reaction conditions:

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

Second, experimental results

Expression patterns of GmNFYA7 at different phosphorus levels As shown in FIG. 1, the GmNFYA7 gene was induced to be expressed in the leaf or root of soybean under low-phosphorus treatment.

Example 2 construction of GmNFYA7 Gene overexpression vector (gateway method)

First, experiment method

1. Primer design

Downloading CDS sequence (nucleotide sequence is shown as SEQ ID NO: 3) of GmNFYA7 gene from a website, designing a specific amplification primer thereof,

OE-GmNFYA7.F：

gggacaagtttgtacaaaaaagcaggcttcTCATGTGTGTAGATTGGGACACTG；

OE-GmNFYA7.R：

ggggaccactttgtacaagaaagctgggtcCTTAGGATGTTCTATCTGATGGTGC。

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-GmNFYA7, 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:

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 load connection System

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.

TABLE 6 purpose Carrier attachment System

Second, experimental results

The GmNFYA7 gene overexpression vector is 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

The Agrobacterium GV3101 stock solution carrying the target vector (the GmNFYA7 gene overexpression vector constructed in example 2) was subjected to streaking activation, and first inoculated onto 50mL of YEP (Kan + Rif) culture plate and cultured at 28 ℃ for 2 days; after the growth of the bacteria, single clone is picked up, the bacteria are shaken in a small tube to obtain 2mLYEP (Kan + Rif) liquid, and then the agrobacterium liquid is subjected to amplification culture in 500mL of YEP (Kan + Rif) liquid and cultured at the temperature of 28 ℃ and the rpm of 200 for 20 hours. The broth was centrifuged at 4000rpm for 10 minutes, the supernatant was discarded, and the Agrobacterium was resuspended in one volume of fresh infiltration 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 high 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 formulations are 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 1/2MS culture medium (containing hygromycin multiplied by 1000), 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 is possible to be 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, RNA was extracted using an RNA extraction kit, and further reverse-transcribed into cDNA, and finally the relative gene expression level was determined by fluorescent quantitative PCR (as described in example 1).

Second, experimental results

The relative expression amount of the GmNFYA7 gene is shown in FIG. 2, the expression level of GmNFYA7 in the strain over-expressing GmNFYA7#5, #6, #20 is obviously higher than that of the wild type, wherein the strains of OE-GmNFYA7#5, OE-GmNFYA7#6 and OE-GmNFYA7#20 are 1192 times, 5602 times and 4875 times of the wild type respectively, and statistical analysis shows that the difference reaches a significant level (, rho <0.05), a very significant level (, rho <0.01) and a very significant level (, rho < 0.001).

Example 4 excess GmNF-YA7 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), 20 samples each treated, three biological replicates, and 7 days of growth. Measurement indexes are as follows: lateral root number, main root length, lateral root density, lateral root number of breakthrough epidermis.

Second, experimental results

FIGS. 3 to 5 show the effect of excess GmNFYA7 on the root system of transgenic Arabidopsis plants after 7 days of Arabidopsis growth. As shown, the number of lateral roots of the OE-GmNFYA7 strain (#5, #20) was significantly increased compared to the wild type under normal and low phosphorus conditions; the main root length and the lateral root density are not obviously different from the wild type; under low phosphorus stress, the lateral root number of OE-GmNFYA7 strain (#5, #20) that breached the epidermis was significantly higher than the control. The result shows that the GmNFYA7 gene obviously promotes lateral root primordium to break through the epidermis under the low-phosphorus condition, and the number of the lateral roots is increased.

Sequence listing

<110> southern China university of agriculture

<120> soybean low-phosphorus response gene for promoting lateral root formation, protein and application thereof

<160> 4

<170> SIPOSequenceListing 1.0

<210> 1

<211> 5679

<212> DNA

<213> Glycine max

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agctatgatt aataggtgtt tctaaatcaa tttgatatac aattgaattg tcttctgcag 840

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tcaccaaggc acaactttac cactgcacca gggctcgccc tctgtataat aactaagtaa 2700

aaactaagat aatttttttt tgtggttgct gagaaaaatt aattacacta tctctattta 2760

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ttattttctc tccttttttc ttctcttatt tttatttaga tatctctatt cttttttatt 3000

gctgtattga atagctgtcc ttcagtgatt ttaaatgact gaaagtcgtt agtttaagat 3060

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agttagtctg ccatgctatc taaagtggtg tttttcttaa agtgatgcat ttgactagtc 3720

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tcacagcttg gaactgggca tgctgtggta tgaacttcac attttaatgg gccatcttat 3960

cattgtctta gtaaacaata tatgttttgg tttgccatcc tcagtgctct cattggagct 4020

ggtatctcag taaatgctta ggaatgtttt gttttgtatt ggtcataggc tcagcctttt 4080

gatgtcatca aataatgaag ttttgtccga attttgctgt tttccttgga ggttttgttt 4140

gatgatgaaa ttattttctt tttaaataac cctagttttt tttacctgac aattgaaggt 4200

tagtgaatga ttgcctgagc aagtactggt gccagttgtt atcatgttat gttgatgtaa 4260

tataaagaag taagaagtgg ccgctcagac ttggtcaggc aattgaatta cttctttgtc 4320

ccattggccc tcctggattt tcttatctca ttgctgttgg tttttctgtc ctgtcatatt 4380

ttagaaaata aactgtgttt gaactttact gatttttgtt gcttggtgta tctaattgaa 4440

ggtaccgccc acttacccat atccagatcc atactacaga agtatctttg ctccctatga 4500

tgcacaaact tatcccccac aaccctatgg tggaaatcca atggttcatt ctaatctttt 4560

gttgttgatg tttgcaatat agtgtttctt gttatgttcc tcactattga atcgtttcct 4620

gtgccaggtc caccttcagt taatgggaat tcaacaagca ggtgttcctt tgccaactga 4680

tacagttgag gagcctgtgt ttgtcaatgc aaaacagtat catggtatat taagacgcag 4740

acagtcccgt gctaaagctg aatcagaaaa aaaggctgca aggaatcgga aggtatatcc 4800

ctacccctat ctgttctcat gttcctcttt aataatgcga aaagatgatt aagaaaaact 4860

aattgttaaa aaaggaaaac gaagggaaga aggttcctgg aagttgtgtt gttagctgct 4920

aacttctaga tgttgatatt tttgtgtgtg cctacatgga tttgtttgac tcacaaactt 4980

ccatgcatcc ttcatggatt tacttgatcc ttaaacaaat gtttcagcca tacttgcatg 5040

aatctcgaca tttgcatgca ctgagaagag caagaggatg tggaggtcgg tttttgaatt 5100

caaagaaaga tgagaatcaa caggatgagg ttgcatcaac tgacgaatca cagtccacta 5160

tcaatctcaa ttctgataaa aatgagcttg caccatcaga tagaacatcc taaaactaca 5220

gaaatggtga tgctgtagat tgcagggatc tgttgtgtat atctatattg ggagatgaat 5280

ctccaaccaa cagtatcctc agatatctcc ctattattca ttctgtcgta caacgccata 5340

ggtataagta taggttgtgt agtaggtatg ttaggaggtt gcaaaataaa acaagtaaaa 5400

tgtaaattga agtgattcaa ctaagtctat ccccaatgtg gtcctttctt gcctttttag 5460

gtatttttat tgtgtgggct tttctttgta ttatttggtg cctctgaggg aaagagaaga 5520

gattatccga atgcgtgatg gtttggttgg atcatgttta tttagcttgt gagaattaat 5580

tgttgtagca ctcttttgat ttgttgttat aaacgaggat gaaaagttta aaccaggatt 5640

ggtcaagttc cgcacagttg cagatagaag gaagaggtt 5679

<210> 2

<211> 1452

<212> DNA

<213> Glycine max

<400> 2

gttaagtttg cctttatttg tgtgggtggg tgtggtgtgg ataggtggca gctacgacac 60

aacacaacac aacacaacac aacacagtcc taagttgtaa gaaacactct cttctccttt 120

ctcactattg ttctgttact gttttttgca gcaacacttc agttcaatta acgaactaca 180

ccactttctt tctcttcttc gactgctctg taaccgaaaa cctccctttc ccagtttcga 240

atcttttgtt tctgcctttg gttactgttt ttccgagcca tgctattcat tattgtcctt 300

cgaatcggat tggtaacaag gaatttatgt gtgtagattg ggacactgta ttgcatgtaa 360

atcaggaaat catgacttct actcatgacc tctcagataa tgaagctgat gaccagcagc 420

agtcggaatc acaaatggag cctttatctg caaatggaat ttcttatgca ggtattgcta 480

ctcagaatgt tcagtatgca acaccttcac agcttggaac tgggcatgct gtggtaccgc 540

ccacttaccc atatccagat ccatactaca gaagtatctt tgctccctat gatgcacaaa 600

cttatccccc acaaccctat ggtggaaatc caatggtcca ccttcagtta atgggaattc 660

aacaagcagg tgttcctttg ccaactgata cagttgagga gcctgtgttt gtcaatgcaa 720

aacagtatca tggtatatta agacgcagac agtcccgtgc taaagctgaa tcagaaaaaa 780

aggctgcaag gaatcggaag ccatacttgc atgaatctcg acatttgcat gcactgagaa 840

gagcaagagg atgtggaggt cggtttttga attcaaagaa agatgagaat caacaggatg 900

aggttgcatc aactgacgaa tcacagtcca ctatcaatct caattctgat aaaaatgagc 960

ttgcaccatc agatagaaca tcctaaaact acagaaatgg tgatgctgta gattgcaggg 1020

atctgttgtg tatatctata ttgggagatg aatctccaac caacagtatc ctcagatatc 1080

tccctattat tcattctgtc gtacaacgcc ataggtataa gtataggttg tgtagtaggt 1140

atgttaggag gttgcaaaat aaaacaagta aaatgtaaat tgaagtgatt caactaagtc 1200

tatccccaat gtggtccttt cttgcctttt taggtatttt tattgtgtgg gcttttcttt 1260

gtattatttg gtgcctctga gggaaagaga agagattatc cgaatgcgtg atggtttggt 1320

tggatcatgt ttatttagct tgtgagaatt aattgttgta gcactctttt gatttgttgt 1380

tataaacgag gatgaaaagt ttaaaccagg attggtcaag ttccgcacag ttgcagatag 1440

aaggaagagg tt 1452

<210> 3

<211> 660

<212> DNA

<213> Glycine max

<400> 3

atgtgtgtag attgggacac tgtattgcat gtaaatcagg aaatcatgac ttctactcat 60

gacctctcag ataatgaagc tgatgaccag cagcagtcgg aatcacaaat ggagccttta 120

tctgcaaatg gaatttctta tgcaggtatt gctactcaga atgttcagta tgcaacacct 180

tcacagcttg gaactgggca tgctgtggta ccgcccactt acccatatcc agatccatac 240

tacagaagta tctttgctcc ctatgatgca caaacttatc ccccacaacc ctatggtgga 300

aatccaatgg tccaccttca gttaatggga attcaacaag caggtgttcc tttgccaact 360

gatacagttg aggagcctgt gtttgtcaat gcaaaacagt atcatggtat attaagacgc 420

agacagtccc gtgctaaagc tgaatcagaa aaaaaggctg caaggaatcg gaagccatac 480

ttgcatgaat ctcgacattt gcatgcactg agaagagcaa gaggatgtgg aggtcggttt 540

ttgaattcaa agaaagatga gaatcaacag gatgaggttg catcaactga cgaatcacag 600

tccactatca atctcaattc tgataaaaat gagcttgcac catcagatag aacatcctaa 660

<210> 4

<211> 219

<212> PRT

<213> Glycine max

<400> 4

Met Cys Val Asp Trp Asp Thr Val Leu His Val Asn Gln Glu Ile Met

1 5 10 15

Thr Ser Thr His Asp Leu Ser Asp Asn Glu Ala Asp Asp Gln Gln Gln

20 25 30

Ser Glu Ser Gln Met Glu Pro Leu Ser Ala Asn Gly Ile Ser Tyr Ala

35 40 45

Gly Ile Ala Thr Gln Asn Val Gln Tyr Ala Thr Pro Ser Gln Leu Gly

50 55 60

Thr Gly His Ala Val Val Pro Pro Thr Tyr Pro Tyr Pro Asp Pro Tyr

65 70 75 80

Tyr Arg Ser Ile Phe Ala Pro Tyr Asp Ala Gln Thr Tyr Pro Pro Gln

85 90 95

Pro Tyr Gly Gly Asn Pro Met Val His Leu Gln Leu Met Gly Ile Gln

100 105 110

Gln Ala Gly Val Pro Leu Pro Thr Asp Thr Val Glu Glu Pro Val Phe

115 120 125

Val Asn Ala Lys Gln Tyr His Gly Ile Leu Arg Arg Arg Gln Ser Arg

130 135 140

Ala Lys Ala Glu Ser Glu Lys Lys Ala Ala Arg Asn Arg Lys Pro Tyr

145 150 155 160

Leu His Glu Ser Arg His Leu His Ala Leu Arg Arg Ala Arg Gly Cys

165 170 175

Gly Gly Arg Phe Leu Asn Ser Lys Lys Asp Glu Asn Gln Gln Asp Glu

180 185 190

Val Ala Ser Thr Asp Glu Ser Gln Ser Thr Ile Asn Leu Asn Ser Asp

195 200 205

Lys Asn Glu Leu Ala Pro Ser Asp Arg Thr Ser

210 215

Claims (7)

1. The application of GmNFYA7 protein in promoting the growth of plant roots is characterized in that the amino acid sequence of GmNFYA7 protein is shown as SEQ ID NO: 4, the promotion of the growth of the plant root system is to increase the number of lateral roots.
2. The application of the GmNF-YA7 gene in promoting the growth of plant roots is characterized in that the nucleotide sequence of the GmNFYA7 gene is shown as SEQ ID NO: 1. SEQ ID NO: 2 or SEQ ID NO: 3, the promotion of the growth of the plant root system is to increase the number of lateral roots.
3. 3. The use of claim 1 or 2, wherein the increase in lateral root numbers is due to the break of lateral root primordia through the epidermis.
4. 4. Use according to claim 1 or 2, wherein the plant is a dicotyledonous plant.
5. The application of GmNFYA7 protein in promoting low phosphorus stress resistance is characterized in that the amino acid sequence of GmNFYA7 protein is shown as SEQ ID NO: 4, respectively.
6. The application of the GmNFYA7 gene in promoting low-phosphorus stress resistance is characterized in that the nucleotide sequence of the GmNFYA7 gene is shown as SEQ ID NO: 1. SEQ ID NO: 2 or SEQ ID NO: 3, respectively.
7. 7. A method for increasing the number of lateral roots of a plant and/or promoting the break through of lateral root primordia into epidermis is characterized in that GmNFYA7 gene and/or GmNFYA7 protein are overexpressed in the plant, and the amino acid sequence of the GmNFYA7 protein is shown as SEQ ID NO: 4, the nucleotide sequence of the GmNFYA7 gene is shown as SEQ ID NO: 1. SEQ ID NO: 2 or SEQ ID NO: 3, respectively.

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