KR101450398B1 - Root-specific promoter, expression vector comprising the same, transformed plants thereby and method for preparation thereof - Google Patents

Abstract

The present invention relates to a promoter specifically expressed in a root, an expression vector containing the promoter, a transformant transformed with the expression vector, and a method for producing the same.
The promoter of the present invention can be used for research for enhancing environmental stress tolerance to the root of major cereals including rice by specifically controlling gene expression necessary for improvement of the trait of rice root, It is expected to be used to improve traits.

Description

Root-Specific Promoters, Expression Vectors Containing the Same, Transgenic Plants by the Same, and Methods for Producing the Transformed Plants

The present invention relates to a promoter specifically expressed in a root, an expression vector containing the promoter, a transformant transformed with the expression vector, and a method for producing the same.

Rice (Oryza sativa) is one of the most important food crops in the world. It has been steadily increasing its yields over the past two decades, but by 2020 it will have to produce more than 70% have. However, the agricultural land where rice is cultivated is getting smaller and smaller as the industrialization phenomenon of each country in the world, and the new genetic resource which is the subject of breeding is depleted and the introduction of the foreign useful gene is required. Fortunately, the development of molecular biology has enabled the isolation and manipulation of exogenous useful genes, transforming useful genes into many plant cells, including rice, to obtain transformants with new genes, Research has become a good source of breeding as well. The production of new varieties using this transformation will not only lead to a high value-added industry in the coming 21st century, but will also solve some of the food problems that are the biggest problem of humanity.

In the 1980s, transgenic plants were obtained by using polyethylene glycol (PEG) and electroporation in protoplasts. In the late 1990s, gene gun utilization became popular, and in recent years, The use of Agrobacterium, which has been widely used in plants, is also widely used. Other methods include pollen pathway and microinjection method.

The promoter can achieve the transformational purpose by locating the expression of the foreign gene only at the whole body of the plant or a specific tissue, and can be classified as follows according to its function.

First, systemic expression-inducible promoters can be mentioned. As a plant systemic expression-inducing promoter, a promoter of 35S RNA gene of cauliflower mosaic virus (CaMV) is used as a typical promoter for dicotyledonous plants. Actin and maize ubiquitin gene promoters have been mainly used as promoters for the expression of the whole plants in rice plants and rice plants. Recently, a promoter of the rice cytochrome C gene (OsOc1) has been developed by domestic researchers, (Cf. Reg. No. 10-0429335). They are already inherent in inducing the expression of antibiotics, herbicide resistance genes and reporter genes used as selection markers in the plant transformation basic carrier. In the research aspect, Promoters considered. Because these promoters lack tissue-selectivity or organ-selectivity, transgenic selectable markers and the like are expressed in whole plants, resulting in delayed plant growth. In addition, due to the nature of the promoter, the expression of the introduced gene is limited to the target organ, and can not be expressed sufficiently, resulting in an inferior economic efficiency of the transformant.

Second, seed-specific promoters can be mentioned. As a representative example, the rice glutelin promoter used for the development of golden rice as promoters of rice major storage protein gene has been widely used to induce seed-specific expression of monocotyledonous plants so far. Promoters that are mainly used to induce seed-specific expression include soybean-derived lectin promoter, cabbage-derived napin promoter and γ-tocopherol methyl transferase (γ-TMT) in Arabidopsis seeds. A patent application (Patent Application No. 10-2006-0000783) for seed-specific expression induction of carrot-derived DC-3 promoter and perilla derived oleosin promoter used in the study promoting the production of vitamin E by inducing gene expression . The seed-specific promoters are mainly used for the purpose of accumulating useful proteins and producing beneficial substances in major crops in which seed itself is used as a food, a food, or a raw material for food.

Third, other tissue specific promoters such as leaves may be mentioned. (RbcS: ribulose bisphosphate carboxylase / oxygenase small subunit) promoter which induces expression of a strong gene only in photosynthetic tissues such as leaves, RolD promoter inducing expression of plant roots derived from Agrobacterium, potato-derived tuber A specific expression-inducible patatin promoter, and a tomato-derived fruit maturation-specific expression-inducing PDS (phytoene synthase) promoter.

Fourth is root specific expression promoter. (IbMADS) and sugar-induced adipose-glucose pyrophosphatase (AGPase) genes were isolated from the sweet potato-derived rice paddy rice, and the peroxidase (prxEa) (Korean Registration Nos. 10-0604186 and 10-0604191), confirming that the promoter induces specific expression in roots and induces root-specific transient expression in carrot and radish.

In addition, although the development is currently underway, new promoters capable of regulating the location of gene expression more precisely according to the intention of the developer, for example, genes that should be expressed only in specific organs (petals, roots, leaves, stems, etc.) , Infertility, burning, specific metabolism-related substances, defensive substances, etc.), there is a need to continue to develop promoters that operate only at specific institutions or periods.

(US) 10-2008-0048817 (US) 10-2008-0067325

The present invention is intended to provide a promoter capable of expressing the expression of a foreign gene locally only to a root, not a whole body of the plant, an expression vector containing the promoter, a transformant transformed with the expression vector, and a method for producing the same.

In order to achieve the above object, the inventors of the present invention have discovered that promoters capable of regulating plant tissue or organ-specific gene expression can be identified, thereby specifically expressing or inhibiting a gene of interest, And to establish a system with In this process, it was confirmed that the promoter of LOC_Os10g31680 of rice was not expressed in petals, leaves, leaves, stomachs, stamens, seeds, etc., but was specifically expressed in roots. Thus, the expression vector containing the promoter was obtained The present inventors have completed the present invention by confirming that a reporter gene is expressed in a root-specific manner in a transformant transformed by introducing it into a plant.

In more detail, we collected 983 species of published rice affymetrix data and reclassified them into 17 organelles. The affymetrix array allows the analysis of gene expression represented by 57,000 probes in a single experiment and the organ-specific expression pattern produced by reconstruction of the large-scale collection of affymetrix arrays is quite high Reproducibility. Through analysis of the expression pattern of this data, various organ and tissue specific genes were isolated. Among them, the present inventors have been interested in finding promoters necessary for expression of important traits in roots.

Through the above method, 84 kinds of genes showed high expression patterns in roots as compared with other genes.

The gene expression pattern regulated by these genes was precisely analyzed by expressing the GUS reporter gene in the 3 'direction of these promoters and measuring its activity in the plant.

Based on the nucleotide sequencing information, the promoter up to 1,969 bp in front of ATG of LOC_Os10g31680 was isolated by genomic DNA PCR and cloned into pGEM T-Ease vector to decode the base sequence.

The thus isolated promoter was inserted into pGA3383 vector to prepare a transformed plant. By verifying the GUS response in the transformed plants, it was found that root-specific gene expression could be induced using the corresponding promoter.

The present invention provides a root-specific promoter (hereinafter referred to as "the promoter of the present invention") that induces the expression of foreign genes to be specific to roots, as a promoter of LOC_Os10g31680 of rice.

The size of the genomic DNA of LOC_Os10g31680 is 717 bp, coding for glycine-rich cell wall structural protein 2 and consists of one exon. LOC_Os10g31680 is translated into 191 amino acids to make proteins. LOC_Os10g31680 exists on chromosome 10 long arm. The LOC_Os10g31680 sequence is shown in SEQ ID NO: 2.

That is, the present invention provides a promoter for root-specific expression of a foreign gene, including the promoter of LOC_Os10g31680 of rice, which is the nucleotide sequence shown in SEQ ID NO: 1.

A promoter is an essential element to regulate environmental, temporal, and systemic expression of a gene. Root-specific promoters according to the present invention can be used to induce root-specific expression sites of foreign useful genes in plants.

The invention also provides a vector comprising said promoter and operatively linked foreign genes.

The term "vector" refers to a DNA fragment (s), nucleic acid molecule, which transfers into a cell, which can be cloned and independently reprogrammed in host cells. "Expression vector" means a recombinant DNA molecule comprising a desired coding sequence and a suitable nucleic acid sequence necessary for expressing a coding sequence operably linked in a particular host organism. The expression vector may generally be derived from a plasmid or viral DNA, or may contain both elements. Thus, an expression vector refers to a recombinant DNA or RNA construct, such as a plasmid, phage, recombinant virus, or other vector that, upon introduction into an appropriate host cell, results in the expression of the cloned DNA. Suitable expression vectors are well known to those skilled in the art and include those that replicate in eukaryotic and / or prokaryotic cells and those that remain as episomes or that are integrated into the host cell genome.

The above-mentioned normal vector may be any vector capable of introducing the promoter of the present invention, preferably a vector derived from a Ti-plasmid such as a pCAMBIA family or a pGA family vector such as pGA3383, pCAMBIA1381, pCAMBIA1391, pGWB3 And the like. More preferably, pGA3383 can be used.

The expression vector of the present invention includes a nucleic acid consisting of the nucleotide sequence shown in SEQ ID NO: 1 and a functionally equivalent fragment thereof.

Means a fragment or a fragment of a nucleic acid consisting of the nucleotide sequence shown in SEQ ID NO: 1, which exhibits substantially equivalent effect to the promoter of the present invention. Such a nucleic acid fragment has a homology of 98% or more with respect to the nucleotide sequence shown in SEQ ID NO: 1, and such a nucleic acid fragment can be easily produced by molecular biological methods well known in the art.

The expression vector of the present invention allows the gene for the target protein to be expressed in the 3 'direction.

Preferably, the expression vector may comprise a useful foreign gene that is specifically intended to be expressed at the root.

The foreign gene may be anything related to the characteristics of roots such as root generation, development, and the like. In order for plants to grow, the absorption of nutrients and water is important, and the root is responsible for its role. It also secures rooted plants and synthesizes secondary metabolites. Examples of genes related to root hair development include ROOT HAIRLESS1 and EXPA17 of rice, and CROWN ROOTLESS1 of rice , ROOTLESS CONCERNING CROWN AND SEMINAL ROOTS of corn , and the like, which are associated with tubal development. In addition, the present expression vector can be used for root-specific expression of foreign genes such as genes responsive to stress such as OsNAC10 .

The above expression vector may be one in which the promoter of the present invention is introduced into a conventional vector, and the expression vector may be prepared by introducing the promoter of the present invention into such a manner as will be readily apparent to those skilled in the art. It is possible.

The present invention also provides a transformed cell transformed with said vector.

Such recombinant vectors may be introduced into host cells cultured using well known techniques such as infection, transfection, transfection, electroporation and transformation. Representative examples of the host include bacterial cells such as E. coli, Streptomyces and Salmonella typhimurium cells; And plant cells.

Any plant cell can be used as "plant cell" used for transformation of a plant. The plant cell may be any type of cultured cell, cultured tissue, cultured organ or whole plant, preferably cultured cell, cultured tissue or culture organ, and more preferably cultured cell. "Plant tissue" refers to a tissue of a differentiated or undifferentiated plant, such as, but not limited to, stem cells, leaves, cancer tissues, and various types of cells used in culture, such as single cells, protoplasts, Callus tissue.

The present invention also provides a transformed transgenic plant transfected with said vector or said transformed cell.

These transgenic monocotyledonous plants exhibit the characteristic that the introduced foreign gene is specifically expressed in the root.

"Transgenic monocotyledonous plants of the present invention" and "functionally equivalent transgenic plants" use a nucleotide sequence having 98% or more sequence homology, as compared to a nucleic acid sequence comprising the nucleotide sequence shown in SEQ ID NO: And the specific promoter effect in the root is substantially equivalent to that of the transgenic plant.

Transformation of a plant of the present invention can be carried out by any method known in the art for transferring DNA to a plant. Such transformation methods do not necessarily have a regeneration and / or tissue culture period. Transformation of plant species is now common for plant species, including both terminal plants as well as dicotyledonous plants. For example, the calcium / polyethylene glycol method for protoplasts (Krens, FA et al., 1982, Nature 296, 72-74; Negrutiu I. et al., June 1987, Plant Mol. Biol. 8, 363-373) (Shillito RD et al., 1985 Bio / Technol. 3, 1099-1102), microinjection into plant elements (Crossway A. et al., 1986, Mol. Gen. Genet. (Klein et al., 1987, Nature 327, 70), infiltration of plants or mature pollen of various plant elements (DNA or RNA-coated) Infections caused by (non-integrative) viruses in Agrobacterium tumefaciens mediated gene transfer (EP 0 301 316), and the like.

The terminal plant to be transformed in the present invention can be used without limitation in the terminal plant including the rice (Oryza sativa L.) used in the present embodiment.

Preferably rice (Oryza sativa L.) is used.

Such mutants can be easily produced by molecular biological methods well known in the art.

The present invention also provides a method for transgenic monocotyledonous plants comprising transforming monocotyledonous plants with the vectors and expressing the foreign genes in monocotyledonous plants.

Preferably, the method comprises the steps of: preparing a vector comprising a promoter comprising the nucleotide sequence shown in SEQ ID NO: 1 and a foreign gene operatively linked thereto; Introducing the expression vector into rice; And a step of selecting a rice transformant to which the expression vector is introduced and which is specifically expressed in the roots of rice.

The promoter of the present invention is a promoter that induces specific expression in roots and can be used in research for enhancing tolerance to environmental stress on the roots of major grains including rice, by specifically controlling the gene expression necessary for improvement of root quality And can be used to improve the agronomic traits of root crops.

FIG. 1 is a heatmap showing the isolated gene showing preferential expression in the root and its expression pattern. FIG.
Figure 2 is a diagram showing microarray results showing preferential expression in the roots of the LOC_Os10g31680 gene.
Figure 3 shows a diagram of the vector used for the production of the LOC_Os10g31680 promoter GUS plant.
Fig. 4 shows root-specific expression of the LOC_Os10g31680 promoter in transgenic rice through GUS staining.

Hereinafter, the present invention will be described in more detail with reference to examples. However, the following embodiments are only examples for helping understanding of the invention, and thus the scope of the present invention is not limited thereto.

< Example  1> Large selection of rice genes specifically expressed in roots

In order to screen for genes specifically expressing in the root, 983 species of rice epimetric microarrays collected from the open microarray database NCBI GEO (http://www.ncbi.nlm.nih.gov/geo/) (affymetrix microarray) data were reconstructed by 17 organizations / organizations to construct a database for gene expression analysis.

This is shown in Table 1.

Table 1. Database for gene expression analysis reconstructed by institution / organization

Using the K-means clustering analysis method, gene groups having high expression in the root of the database for gene expression analysis shown in Table 1 were isolated.

This was performed using MeV's K-means clustering analysis method for microarray data analysis. 84 genes with preferential expression in the root of the database were analyzed.

The results are shown in Fig.

As shown in Fig. 1, a gene which shows stronger expression toward yellow is closer to blue, and the expression becomes lower toward blue. In other words, we identified 84 genes that showed preferential expression in the root among 17 reconstructed gene databases.

< Example  2> Through microarray data analysis LOC _ Os10g31680  Expression analysis of genes by tissue

The expression pattern of LOC_Os10g31680 gene was analyzed by microarray database analysis and the results are shown in FIG.

As shown in FIG. 2, the tissue-specific expression of the LOC_Os10g31680 gene can be confirmed to be strong in the root, which means that expression in the root is preferential.

< Example  3> promoter for root-first expression Cloning

PCR was performed using the primers prepared to clone the promoter of LOC_Os10g31680 and the genomic DNA of Oryza sativa L. japonica cultivar-group. Template DNA was used at 100 ng and each primer was used at 5 pmol. PCR conditions were 5 min at 95 ° C; Repeating 4 times at 95 캜 for 30 seconds, at 55 캜 for 30 seconds and at 68 캜 for 300 seconds; 30 seconds at 95 ° C, 30 seconds at 62 ° C, and 300 seconds at 68 ° C for 35 times and finally 68 ° C for 10 minutes. Primer for promoter cloning is shown in Table 2 below.

Table 2. Primers for cloning promoter of LOC_Os10g31680

The PCR product was electrophoresed to confirm its size and cloned into a pGEM T-EG vector and sequenced. The cloned DNA was digested with BamH1 and Hpa1, followed by ligation with pGA3383 vector (vector for promoter-GUS cloning) digested with the same restriction enzyme at 14 DEG C for 12 hours. The ligation product was mixed with 50 μl of Top10 E. coli competent cells, transferred to a 1.5 ml tube, and left on ice for 15 minutes. Subsequently, the plate was allowed to stand in a 37 ° C oven for 1 minute, and then 1 ml of LB liquid medium was further placed in the tube and left in a 37 ° C shaking incubator for 3 hours. Then, the cells were plated on tetracycline-resistant LB solid medium and the resulting colonies were awaited for 12 hours and then mini-prepared after cell culture in 1 mL of LB liquid medium. DNA was digested with restriction enzymes BamHI and HpaI, and the band was identified by electrophoresis of agarose gel. The cloning DNA thus obtained was subjected to base sequence analysis again to select DNA that did not cause an error. A diagram of the vector used for the production of the LOC_Os10g31680 promoter GUS plant thus constructed is shown in FIG. Nucleotide sequencing was performed by capillary sequencing (Macrogen). Primers were used for nucleotide sequence analysis using NGUS1, RB (right border), and sequences in the promoter.

Table 3. Primers for sequencing

The results of the analysis are shown in Sequence Listing 1. The promoter corresponding to the 1,969 bp promoter sequence precedes the ATG of the LOC_Os10g31680 gene corresponds to a promoter preferentially expressed in the root.

Example 4 Production of Transformed Cells

The plant expression vector prepared in Example 3 was extracted.

50 아 of Agrobacterium tumefaciens LB4404 and 2 의 of plant expression vector were mixed and left on ice for 15 minutes. Subsequently, the substrate was placed in liquid nitrogen for 75 seconds. Then, the substrate was allowed to stand in an oven at 37 ° C for 5 minutes. Then, 1 ml of LB liquid medium was added thereto, followed by incubation in a 28 ° C shaking incubator for 3 hours. Next, the cells were plated on a tetracycline-resistant LB solid medium and incubated for 36 hours. The resulting colonies were cultured in 1 mL of LB liquid medium, mini-preparations were performed, and BamHI and HpaI were used for enzyme cleavage.

The resulting transformed Agrobacterium was used for transgenic rice transformation experiments.

Example 5 Production of Transgenic Plants

In the N6D solid medium, Dongjinbyeon seeds were grown in a 28 ℃ growth chamber for 7 days to produce calli of rice. The resulting callus was mixed with the cells cultured for 72 hours with Agrobacterium transformed with the plant expression vector obtained in Example 4 and left in a 22 占 폚 cancer-treated growth chamber in a medium containing N6D-Acetosyringone.

The callus contaminated with Agrobacterium was rinsed 5 times with tertiary distilled water. Subsequently hygromycin selection was performed in N6D solid medium (hygromycin 30 mg / L) and subculture (hygromycin 40 mg / L) . Selection was made in the 28 ℃ growth room for 2 weeks each for 4 weeks. The divided callus was transferred to the MSR (hygromycin 40 mg / L) solid medium for regeneration and induced to regenerate in a light condition growth chamber for 4 weeks at 28 ° C. The plants were transferred to MS solid medium, grown in a light condition growth room for 7 days, To grow regenerated plants.

Example 6 Expression of Root-Priority Expression Using Transgenic Plants

In order to confirm the organ specific GUS expression of regenerated plants, 10 seeds harvested were cultured at 28 ° C for 7 days and all individuals were stained with GUS.

The GUS solution was prepared by mixing 200 ml of 0.5 M sodium phosphate buffer, 100 ml of 12.5 mM potassium ferricyanide, 100 ml of 12.5 mM potassium ferrocyanide, 20 ml of 0.5 M EDTA, 50 ml of 10% Triton X-100, 1 g of X-glucin dissolved in 10 ml of DMSO, 200 ml of distilled water was added to make 1 L volume. Each plant was treated with 15 ml of GUS staining solution for 24 hours, followed by decolorization at room temperature using 70% ethanol and 100% ethanol sequentially.

As shown in Fig. 4, the transgenic plants can be stained blue only in the roots, which shows predominant GUS expression in the roots. GUS expression is observed around the center of the root and strong GUS expression is observed at the growth point. However, GUS expression in mature leaves, stems, and flowers is not observed.

Based on the results of FIGS. 2 to 4, it was confirmed that the promoter of LOC_Os10g31680 showed preferential expression in the root.

<110> University-Industry Cooperation Group of Kyung Hee University Root-specific promoter, expression vector comprising the same,          transformed plants by and method for preparation thereof <130> P12-084-KHU <160> 7 <170> Kopatentin 2.0 <210> 1 <211> 1969 <212> DNA <213> Oryza sativa <220> <221> promoter <222> (1) .. (1969) <223> Promoter of LOC_Os10g31680 <400> 1 cagatgtaat gaagcacatc gaacggacat gttagaggta atggacagag ctggccatat 60 agcaaactac tggagctaaa tagatcctgt ggttcttagt tggatcttga cttctcacat 120 gcagacacat tatgtaagtg ctaatcgatc aactgattac tactacctcc gtttcacaat 180 gtaagtcatt ctaacatttc ccacattcgt attgatgtta atgaatctat atatatatat 240 atatatagat tcattaacat caatacgaat gtgggaaatg ctggaatgac ttacattgtg 300 aaacggagaa agtacttacc agcataatag tataagatta tcttgcattg tggaagtttc 360 gagatttgaa agaagaatgg tgtctactac tagtactatt aatatactcc tccatctcaa 420 aatatatcac gctagtacta gttgagacat actactctgt caaaaaaaaa aaggcaaact 480 ctagctacga atttggacac gtatatatcc aaattcgtat gtaggattag actttctttt 540 cgaacagagg ggtatatatc ataagactta tagtacgtac taggatatat atgtttcatc 600 cggtactagg ttgatatgtg tagtactgta ctgtaacaca gggcatactt ggatgttttg 660 ctgacatttc tgattgttca gaatgcatac agtttcagta atcattaata ctgggtcgat 720 agagatttga agcaatatta ctgatttttc acagctaatt aggactgatc tactggctat 780 agtgatatta ttgcaaagta ctacagcgat ttctttagtt tgcttccaga ccatcgatct 840 gcaatttccg tttcttttac ctagtggtac caatgtacat tctatacaaa tatctatata 900 tatcgcaatc catgaacatt attagtaaaa cgattatgtc agcgcgtctc gctcgtccca 960 atataatctc ctcgggcaga tcatgcagca gcagccatat gcatattcat cggctgtgtg 1020 ttgtttcttg tatcctgtgc gtggtcgggt taatttgtca gtctgtcact tacaaactgc 1080 gtttgaatgt ttcaaagtca gagctgcaaa agcgatggga tcgagagcct cgaagtcgtc 1140 gtctctcagt attgtcaatt cttctgctag ctgtgtaaca atattcatct gtaatagtag 1200 cagatgcttg tgctgctttg cgcaagtcac actgacacac taagagttaa ttaaactctt 1260 cactctcgta ggcgatctcg atcttaacat gctcatgact agcttaactc agttatacta 1320 ctctctctca aaacaaaaaa aaaaaaagta aactctagct acgaatctga acacatgcat 1380 gtccagtttt ataggtagga ttgtactttt tttttttttg ggacggaggg gtaggtgttt 1440 gaattctcac ccagagccgt gaaagaacaa aattttgaga tatactggga gctctcttga 1500 ttcaaacata taactatata ttttggtata tataacattg gttgagaact taatttaatt 1560 tgggagtttg tttactgcac atagagaagc tagaactata tattttgagt tccattttct 1620 atcaactcat ttttttgtga tagattagcg catagtaaat aatataatac aaatgcttgc 1680 aattatttag atatgtaatt tagttaggat atatatattt catataaact ctacacaaga 1740 gt; atttaagcct caacttgatc cataactaag atgtcaacta cgaaaaaact aacgtggatg 1860 ctgttaaaaa gctagttgta catatggcct caagcactat aaatacccat gtactccttc 1920 acccaaaaat catctcctcc tgccctcata gttcatagag tttgctagc 1969 <210> 2 <211> 717 <212> DNA <213> Oryza sativa <220> <221> gene &Lt; 222 > (1) .. (717) <223> LOC_Os10g31680 <400> 2 atggcaggca ccaagcttgc agctcttggg ttcgttgtcc tcttgagcat tggactagcc 60 agtgctgcaa gagtagaaag atactccagt tcccaaggtt caggtacagg aggtggagag 120 ggaggaggat atgtgaacgg agggggagct gggaaaggag tcggggctgg ttctggtagc 180 agtaactctt atggtgctca tgcaagtggt ggaggtggcg gcggaggtgg tggttacagc 240 caatacggtg gatcaggatc tggtagcgga tacggcacgg gctcaggctc aagccaaact 300 tcacaaaatg gatactatgg ttatggtggg tcatccagtg ctagcggtgc tggtgttggt 360 ggtggtgccg gacaagccgg tggttactgg ccatccaatg gtcatgggtc cggcagtggt 420 acgggctatg gctctagcac tgctaataac tactactatg gaccatatgc aaatgcaaat 480 gctggtggta atggtggcgg taatggtcaa ggccaatatg gtggcagtgg cggtggtgga 540 ggagctggag ttggatctgg tgacgccagc ccttaatttc cttcgtaaag gaaattttaa 600 aaatggagcc cactatgttt cgtaccatgt ctatgaagta gaatttgttc tcatttattt 660 gtacttgttt catgtgttgt gcaacaagaa ataaagggct tcattgattc agtaaaa 717 <210> 3 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> Primer of Os10g3168 Pro BamH1 <400> 3 ggatcccaga tgtaatgaag cacatcg 27 <210> 4 <211> 27 <212> DNA <213> Artificial Sequence <220> <223> Primer of Os10g3168 Pro Hpa1 <400> 4 gttaacgcta gcaaactcta tgaacta 27 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer of NGUS1 <400> 5 aacgctgatc aattccacag 20 <210> 6 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Primer of RB backbone F2 <400> 6 tcgcacggaa tgccaagca 19 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer of Os10g3168_seq1 <400> 7 gactgatcta ctggctatag 20

Claims (7)

Comprising transfecting a monocotyledonous plant with a vector comprising a promoter comprising the nucleotide sequence shown in SEQ ID NO: 1 and a foreign gene operatively linked thereto, and expressing the foreign gene in the monocotyledonous plant, In a root-specific manner. The method according to claim 1, wherein the method for producing transgenic monocotyledonous plants comprises the following steps:
Preparing a vector comprising a promoter comprising the nucleotide sequence shown in SEQ ID NO: 1 and a foreign gene operatively linked thereto;
Introducing the vector into rice; And
Selecting the transgenic monocotyledonous plants to which the vector is introduced and the foreign gene is expressed specifically in the root. 3. The method according to claim 2, wherein the terminal plant is rice (Oryza sativa). A terminal transgenic plant produced by the method of any one of claims 1 to 3, which expresses a foreign gene in a root-specific manner. The transgenic plant of claim 4, wherein the terminal plant is rice (Oryza sativa). 1. A composition for inducing root-specific expression of a foreign gene, comprising a promoter comprising the nucleotide sequence shown in SEQ ID NO: 1 and a vector comprising an operably linked foreign gene. 1. A composition for inducing root-specific expression of a foreign gene, comprising a promoter comprising the nucleotide sequence shown in SEQ ID NO: 1 and a cell transformed with a vector comprising an operatively linked foreign gene.

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