KR101779030B1 - Molecular marker for selecting plant with enhanced zinc content and uses thereof - Google Patents

Abstract

The present invention relates to a molecular marker for selecting a plant having an increased zinc content and a use thereof. When a molecular marker according to the present invention is applied to a rice plant, the rice plant with increased zinc content, And it is expected to be very useful for reducing the time, cost and effort required for breeding. In addition, the spread of rice cultivars cultivated by the method of the present invention is expected to enable production and supply of high quality rice to farmers and consumers.

Description

[0001] Molecular markers for selecting plants with increased zinc content and uses thereof [0002]

The present invention relates to a molecular marker for screening a plant having an increased zinc content and a use thereof. More particularly, the present invention relates to a method for screening a plant having a wild-type GW2 gene comprising the deleted GW2 gene at position 316 of the nucleotide sequence of SEQ ID NO: An oligonucleotide composed of at least 8 consecutive nucleotides including a 315th base and a 316th base in the nucleotide sequence of SEQ ID NO: 2, or a complementary oligonucleotide thereof, A kit for screening plants having increased zinc content compared to a wild type comprising a primer set for selecting a plant, a probe and a microarray, a set of the primer and a reagent for carrying out an amplification reaction, wherein the zinc content is increased compared to the wild type, Plants with increased zinc content compared to wild type using sets It relates to a method for screening.

One of the goals of using molecular genetics for crop improvement is to foster an excellent new variety by searching, characterizing, isolating and exploiting the necessary useful genes. Over the past decade, a variety of techniques have been developed for the genome research of crops. Among them, DNA marker technology provides a solution to many problems that can not be solved by traditional breeding methods. Analysis of quantitative trait locus (QTL) and search and use of useful genes in wild genomes.

The progress of plant genome analysis technology has enabled the study of genetic mutations involved in quantitative trait loci and has enabled the efficient use of marker-assisted selection (MAS) in the transfer and selection of target genes, Quantitative trait loci analysis (QTLs) uses molecular markers and identifies specific parts of chromosomes that are involved in measurable traits so that the number of genes involved in major traits such as yield, fruit weight, heading, and chromosomes You can get general and direct information about the genetic background of major agricultural traits such as location, influence of individual genes, interaction, interactions between quantitative trait loci and environment, and interactions between quantitative trait loci and genetic background .

Wild rice is an important source of useful genes for improving crop yield, pest resistance, stress resistance and related traits. A range of marker-assisted selection techniques have been found to be effective tools for improving yield, resistance, etc., by exploring wildly useful genes, introducing them into cultivars, and selecting them using markers. Currently, large numbers of genes stored in gene banks around the world are widely used as basic research data in the search for new genes involved in yield and other useful traits.

On the other hand, Korean Patent No. 1255336 discloses rice varieties with increased trace element content and functional foods using the same, and Japanese Patent Laid-Open No. 2011-131023 discloses rice varieties having an increased content of trace elements and which are involved in the absorption or transport of metal complexes such as iron Transporters and their genes. However, the molecular markers for selecting plants with increased zinc content of the present invention and their uses have not yet been mentioned.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and the present inventors have found that GW2 It was also confirmed that the gene is also involved in zinc content regulation. Therefore, GW2 By analyzing the gene, a 316th base in the base sequence of the GW2 gene is deleted and a molecular marker capable of easily selecting a plant having increased zinc content compared to the wild type has been developed and provided, Completed.

In order to solve the above problems, the present invention provides a molecular marker composition for selecting a plant having an increased zinc content as compared to a wild type, wherein the 316th base in the nucleotide sequence of SEQ ID NO: 1 contains a deleted GW2 gene.

In addition, the present invention relates to a method for producing an oligonucleotide comprising an oligonucleotide consisting of 8 or more consecutive nucleotides comprising the 315st base and the 316st base in the nucleotide sequence of SEQ ID NO: 2 or a complementary oligonucleotide thereof, A primer set for selecting a plant, a probe, and a microarray.

In addition, the present invention provides a kit for selecting a plant having increased zinc content as compared to a wild type, comprising the primer set and a reagent for carrying out an amplification reaction.

The present invention also provides a method for screening plants having increased zinc content compared to wild type using the primer set.

When the molecular markers according to the present invention are applied to rice crops, it is possible to efficiently select and cultivate rice crops having an increased zinc content compared to the wild type, so that it is very useful for reducing the time, cost and effort required for breeding It is expected to do. In addition, the spread of rice cultivars cultivated by the method of the present invention is expected to enable production and supply of high quality rice to farmers and consumers.

FIG. 1 is a result (A) of the quantitative trait locus (QTL) of HG101, and is a result (B) of seed size comparison between Hwaseong and HG101. HG101 represents near-isogenic lines (NILs) in which the chromosomes of wild rice ( Oryza grandiglumis ) are partially transferred to the genetic background of the rice flour .
FIG. 2 shows the results of comparing the zinc content (A) and the seed size (B) of brown rice and rice hull in Hwaseong and HG101.
Figure 3 shows the zinc content of brown rice in 11 rice varieties.

In order to accomplish the above object, the present invention provides a method for detecting a deletion of GW2 Wherein the zinc content is increased as compared to the wild type.

In the present invention, SEQ ID NO: 1 represents the nucleotide sequence of the wild type GW2 gene, and the 316th nucleotide represents A (adenine).

Further, the present invention provides a method for producing a wild-type (preferably, homologous) oligonucleotide comprising an oligonucleotide composed of 8 or more consecutive nucleotides including the 315th base (A) and the 316th base (C) in the nucleotide sequence of SEQ ID NO: 2 or a complementary oligonucleotide thereof The present invention provides a primer set for screening plants having increased zinc content.

The nucleotide sequence of SEQ ID NO: 2 of the present invention is GW2 (deleted) in which the A (adenine) at position 316 of the nucleotide sequence of SEQ ID NO: 1 is deleted As a nucleotide sequence of the gene, the 315th base is A (adenine), the 316th base is A (adenine) is missing, and the 317th base is C (cytosine). Therefore, the 315th base is A and the 316th base is C in the nucleotide sequence of SEQ ID NO: 2.

The 316th base in the nucleotide sequence of SEQ ID NO: 1 is the deleted GW2 Since the plant containing the gene is a plant having an increased zinc content compared to the wild type, the plant selection screening primer set for increasing the zinc content compared to the wild type is a primer set capable of detecting the 316th base defect in the nucleotide sequence of SEQ ID NO: . Therefore, when the 316th base in the nucleotide sequence of SEQ ID NO: 1 is deleted, GW2 The 315th base and the 317th base are adjacent to each other due to the deletion of the 316th base in the nucleotide sequence of SEQ ID NO: 1, and the primer set includes the adjacent 315th base and 317th base, that is, the 315th base of SEQ ID NO: (A) and the 316th base (C). Such a primer may comprise an oligonucleotide composed of 8 or more consecutive nucleotides comprising the 315th base (A) and the 316th base (C) in the nucleotide sequence of SEQ ID NO: 2 or a complementary oligonucleotide thereof. When the amplified product is generated by amplifying the target sequence using the primer set, the plant sample is a sample whose zinc content is increased compared to that of the wild type, and if the amplification product is not produced, it can be discriminated as a wild type in which the zinc content is not increased . That is, in the wild type, the 316th base in the nucleotide sequence of SEQ ID NO: 1 remains unremoved and does not undergo primer annealing, so that no amplification product is generated with the primer set of the present invention.

In the present invention, the oligonucleotide may be composed of at least 8 nucleotides, preferably 15 to 50 nucleotides, more preferably 15 to 30 nucleotides, even more preferably 15 to 20 nucleotides , But is not limited thereto.

As used herein, the term "nucleotide" is a deoxyribonucleotide or ribonucleotide present in single-stranded or double-stranded form and includes analogs of natural nucleotides unless specifically stated otherwise (Scheit, Nucleotide Analogs, John Wiley, New York (1980); Uhlman and Peyman, Chemical Reviews, 90: 543-584 (1990)).

In the present invention, a "primer" refers to a single strand oligonucleotide sequence complementary to a nucleic acid strand to be copied, and may serve as a starting point for synthesis of a primer extension product. The length and sequence of the primer should allow the synthesis of the extension product to begin. The specific length and sequence of the primer will depend on the primer usage conditions such as temperature and ionic strength, as well as the complexity of the desired DNA or RNA target.

In the present invention, the oligonucleotide used as a primer may also include a nucleotide analogue such as phosphorothioate, alkylphosphorothioate, or peptide nucleic acid, or And may include an intercalating agent. In addition, primers can incorporate additional features that do not alter the primer properties of primers that serve as a starting point for DNA synthesis. The primer nucleic acid sequence of the present invention may, if necessary, comprise a label that is detectable directly or indirectly by spectroscopic, photochemical, biochemical, immunochemical or chemical means. Examples of labels include enzymes (e.g. horse radish peroxidase (HRP), alkaline phosphatase), radioactive isotopes (e.g., ³² P), fluorescent molecules, chemical groups (e.g. biotin) ). The appropriate length of the primer is determined by the characteristics of the primer to be used, but is usually 15 to 30 bp in length. The primer need not be exactly complementary to the sequence of the template, but should be complementary enough to form a hybrid-complex with the template.

In addition, the present invention relates to a method for producing an oligonucleotide comprising an oligonucleotide consisting of 8 or more consecutive nucleotides comprising the 315st base and the 316st base in the nucleotide sequence of SEQ ID NO: 2 or a complementary oligonucleotide thereof, A probe for screening a plant is provided.

The term "probe" in the present invention refers to a hybridization probe, which means a linear oligomer having a natural or modified monomer or linkage comprising a deoxyribonucleotide and a ribonucleotide capable of sequence-specific binding to the complementary strand of the nucleic acid. The probe of the present invention is an allele-specific probe in which a polymorphic site exists in a nucleic acid fragment derived from two members of the same species and hybridizes to a DNA fragment derived from one member but does not hybridize to a fragment derived from another member . Preferably, the probe may be a single strand, more preferably a deoxyribonucleotide, but is not limited to, for maximum efficiency in hybridization.

In addition, the present invention relates to a method for producing an oligonucleotide comprising an oligonucleotide consisting of 8 or more consecutive nucleotides comprising the 315st base and the 316st base in the nucleotide sequence of SEQ ID NO: 2 or a complementary oligonucleotide thereof, A microarray for plant selection is provided.

In the present invention, a plant selection microarray having an increased zinc content as compared to a wild-type microarray means a microarray having a substrate on which the molecular markers of the present invention are immobilized. The term "microarray" refers to a group of oligonucleotides immobilized on a substrate at a high density. The oligonucleotide groups are immobilized in a predetermined region. Such microarrays are well known in the art. Microarrays are described, for example, in U.S. Patent Nos. 5,445,934 and 5,744,305, the contents of which are incorporated herein by reference.

The term "substrate" refers to any substrate that has hybridization properties and to which the marker can be attached under conditions where the background level of hybridization is kept low. Typically, the substrate may be a microtiter plate, a membrane (e.g., nylon or nitrocellulose), a microsphere (bead), or a chip. Prior to application or immobilization to the membrane, nucleic acid probes can be modified to promote immobilization or improve hybridization efficiency. Such modifications may include homopolymer tailings, coupling with aliphatic groups, different reactive functional groups such as NH2, SH and carboxyl groups, or coupling with biotin, hapten, or proteins.

In addition, the present invention provides a kit for selecting a plant having increased zinc content as compared to a wild type, comprising the primer set and a reagent for carrying out an amplification reaction. The reagents for carrying out the amplification reaction may include, but are not limited to, DNA polymerases, dNTPs, and buffers. The dNTPs include dATP, dCTP, dGTP and dTTP. The DNA polymerase can be a commercially available polymerase such as Taq DNA polymerase and Tth DNA polymerase as a heat-resistant DNA polymerase. In addition, the kit of the present invention may further include a user's manual describing optimal reaction performing conditions. Also, in the present invention, the kit for selecting plants having increased zinc content compared to the wild type may be a kit including the probe or the microarray, but is not limited thereto.

The present invention also relates to a method for isolating genomic DNA from a rice sample,

Amplifying the target sequence by using the separated genomic DNA as a template and performing an amplification reaction using the primer set; And

And detecting the amplification product. The present invention provides a method for screening a plant having increased zinc content compared to a wild-type plant.

In the method of the present invention, since the primer set is a primer set specific to the sequence lacking the 316th base in the nucleotide sequence of SEQ ID NO: 1, when the amplified product is generated by amplifying the target sequence using the primer set, The zinc content is an increased sample compared to the wild type, and if the amplification product is not produced, it can be discriminated as a wild type without increased zinc content.

The method of the present invention comprises isolating genomic DNA from a rice sample. The genomic DNA may be extracted from the sample using phenol / chloroform extraction method, SDS extraction method, CTAB isolation method, or commercially available DNA extraction kit, which are conventionally used in the art.

In the method according to one embodiment of the present invention, the amplification product may include, but is not limited to, the 315th base (A) and the 316th base (C) in the nucleotide sequence of SEQ ID NO: 2. The target sequence can be amplified by performing amplification reaction using the separated genomic DNA as a template and using the primer set of the present invention as a primer. Methods for amplifying a target nucleic acid include polymerase chain reaction (PCR), ligase chain reaction, nucleic acid sequence-based amplification, transcription-based amplification system, Strand displacement amplification or amplification with Q [beta] replicase, or any other suitable method for amplifying nucleic acid molecules known in the art. Among them, PCR is a method of amplifying a target nucleic acid from a pair of primers that specifically bind to a target nucleic acid using a polymerase. Such PCR methods are well known in the art, and commercially available kits may be used.

In the method of the present invention, the amplified target sequence may be labeled with a detectable labeling substance. In one embodiment, the labeling material can be, but is not limited to, a fluorescent, phosphorescent or radioactive substance. When the radioactive isotope such as ³² P or ³⁵ S is added to the PCR reaction solution, the amplification product may be synthesized and the radioactive substance may be incorporated into the amplification product and the amplification product may be labeled as radioactive. The set of one or more oligonucleotide primers used to amplify the target sequence is as described above.

The method of the present invention comprises detecting said amplification product. The detection of the amplification product can be performed by DNA chip, gel electrophoresis, sequencing, radioactivity measurement, fluorescence measurement or phosphorescence measurement, but is not limited thereto. As one method of detecting the amplification product, gel electrophoresis can be performed. Gel electrophoresis can be performed using agarose gel electrophoresis or acrylamide gel electrophoresis depending on the size of the amplification product. In addition, the sequence analysis method can be a Sanger method, micro electrophoresis analysis, pyrosequencing, and nanopore single DNA sequence analysis. Also, in the fluorescence measurement method, Cy-5 or Cy-3 is labeled at the 5'-end of the primer. When PCR is carried out, the target is labeled with a fluorescent label capable of detecting the target sequence. The labeled fluorescence is measured using a fluorescence meter can do. In addition, in the case of performing the PCR, the radioactive isotope such as ³² P or ³⁵ S is added to the PCR reaction solution to mark the amplification product, and then a radioactive measurement device such as a Geiger counter or liquid scintillation counter The radioactivity can be measured using a liquid scintillation counter.

Hereinafter, the present invention will be described in more detail with reference to Examples. It is to be understood by those skilled in the art that these embodiments are merely illustrative of the present invention and that the scope of the present invention is not limited thereto.

Example  One. Quantitative trait Locus ( QTL , quantitative trait loci ) analysis

Wild rice, wild rice ( Oryza grandiglumis , CCDD genome, IRGC Acc. no. 101 154) were right backcrossing the progeny BC5F1 repeatedly hitting hwaseongbyeo the F ₁ produced by the pollen parent, was named HG101 by selecting a system fixed to the phenotypic After these children. HG101 showed near-isogenic lines (NILs) in which the chromosome of wild rice ( Oryza grandiglumis ) was partially transferred to the genetic background of the rice seedlings. In order to investigate the effect of specific chromosomal fragments transferred to HG101 on one trait, 150 F2: 3 lines were cultivated by crossing between HG101 / HG101. 150 strains and their parents were reported to the pavement in 2 repetitions and genetic maps were made and QTL analysis was conducted to investigate the major characteristics such as heading date and the relationship between specific bands and traits of wild rice ( Oryza grandiglumis ). Respectively. As a result, QTLs related to the seed width (gw2) and the seed thickness (gt2) of seeds (tgw2) were detected in the marker RM290 region of chromosome 2 (FIG. 1A). Also, as shown in FIG. 1B, it was confirmed that the seed sizes of Hwaseong and HG101 were different from each other.

Example  2. GW2  Sequence analysis of genes and confirmation of zinc content regulation

In the seeds detected by QTL in Example 1, GW2 The characteristics of the gene have been reported by Chinese researchers. A comparison of genomic and cDNA sequences of the FAZ1 allele, a model of GW2, revealed eight exons and seven introns, and the FAZ1 GW2 allele was predicted to encode 425 polypeptides of about 47 kDa Respectively. A comparison of the nucleotide sequence of WY3 allele, FAZ1 of GW2 and the large rice varieties of seeds, revealed two base substitutions that did not cause amino acid mutations in exons 1 and 8 and a premature stop codon in exon 4 of the WY3 allele premature stop codon). The early stop codon induced truncation of 310 amino acid residues and the remaining protein consisted of 115 polypeptides of 13 kDa (Song et al. 2007. Nature Genetics 39 (5) A QTL for rice grain width and weight encodes a previously unknown RING-type E3 ubiquitin ligase).

Regarding the GW2 gene involved in the seeds analyzed by Chinese researchers, in the present invention, the GW2 To confirm the additional function of the gene, the trace element content was measured. In order to investigate the zinc content, brown rice and rice hull were separated from the paddy harvested from the pavement. 200 lips per sample were collected, separated into brown rice and rice husks, weighed, and then decomposed into 65% nitric acid at 160 ° C. The zinc content was measured by inductively coupled plasma mass spectrometry (ICP-MS; Perkin Elmer, Wellesley, MA, USA). As a result, it was confirmed that there was a difference in zinc content between the two lines (Fig. 2). In the grain, HG101 (gw2 ql) was 34 mg per kg of seed, which was about 22% higher than that of 28 mg of HS, and husk was about 34% higher than that of husk. Suggesting that the transfer efficiency is higher in HG101. In addition, GW2 To investigate whether the gene regulates zinc content simultaneously, we selected 56 strains of 150 strains, similar to those of Hwasung, and identified differences in zinc content between RM290 genotypes closely related to GW2 . As a result, it was confirmed that the difference in zinc content according to the marker genotype was highly significant as shown in Table 1 (P <0.02).

Comparison of zinc content of RM12813 by genotype chromosome Marker Characteristic P Feature Average HH ^@ GG ^@ 2 RM12813 Zn content 0.02 25.8 (21) ^&amp; 28.8 (27)

@: Relative phenotypic mean value of HG101 homozygotes, &: Number of lines in each genotype class.

As a result of analyzing the nucleotide sequence of HG101, it was judged that the seed and zinc contents were controlled by the same mechanism, as the Chinese researchers showed the same mutation as the nucleotide sequence of WY3 . There are a variety of different genetic resources in the world, and Chinese researchers have reported that JZ1560, the antagonistic rice, also exhibits the same base mutation as WY3 in the GW2 gene (Ying et al., 2012. Journal of Genetics and Genomics 39: 325-333) . As a result of investigation of zinc content of two kinds of confounding rice (JZ1581 and SLG1) and five kinds of domestic breeding species (Hwa Seon, Drachan, Hwasung, And SLG1 showed a statistically significant higher zinc content than domesticated species (Fig. 3). This result shows that GW2 gene is involved in the regulation of zinc content while being involved in seeds.

Therefore, the later lines such as HG101 cultivated in the present invention may be used as an important crossbreeding example for breeding a system in which the zinc content is increased in the future.

<110> The Industry & Academic Cooperation in Chungnam National University (IAC) <120> Molecular marker for selecting plant with reduced zinc content          and uses <130> PN16381 <160> 2 <170> KoPatentin 3.0 <210> 1 <211> 1278 <212> DNA <213> Oryza sativa <400> 1 atggggaaca ggataggggg gaggaggaag gcgggggtgg aggagaggta cacgaggccg 60 caggggctgt acgagcacag ggacatcgac cagaagaagc tccggaagct gatcctcgag 120 gccaagctcg cgccgtgcta catgggcgcc gacgacgccg ccgccgccgc cgacctcgag 180 gagtgcccca tctgcttcct gtactaccca agtcttaacc gatcaaagtg ttgctcaaaa 240 gggatatgca ccgagtgctt tctccaaatg aaaccaactc acactgctca gcctacacaa 300 tgtccattct gcaaaactcc cagttatgct gtggagtatc gtggtgtaaa gacaaaggag 360 gaaaggagca tagaacaatt tgaagagcag aaagtcatag aagcacaaat gaggatgcgc 420 cagcaagcac ttcaagatga agaagataag atgaaaagaa aacagaacag gtgctcttct 480 agcagaacaa tcacaccgac caaagaagtg gagtatagag atatttgcag cacatccttt 540 tcagtgccgt cataccgatg tgctgagcaa gaaactgaat gctgttcatc ggaaccttca 600 tgctctgccc agactagcat gcgccctttc cattctaggc ataaccgtga tgataacatt 660 gacatgaata tagaggatat gatggttatg gaagcgattt ggcgttccat tcaggagcag 720 ggaagtatag ggaatcctgt ctgtggcaac tttatgcctg taactgagcc atctccgcgt 780 gaacgccagc cattcgttcc agctgcttct ctagaaatac ctcatggtgg tggattttcc 840 tgtgcggttg cggcaatggc tgagcaccag ccacccagta tggacttctc ttacatggct 900 ggcagcagcg cattcccagt tttcgacatg ttccggcgac catgcaacat tgctggtgga 960 agcatgtgta atctggagag ctcaccggag agctggagcg ggatagcacc aagctgcagc 1020 agggaagtgg taagagaaga aggagagtgc tcggctgacc actggtcgga gggtgcagag 1080 gccggaacaa gctacgcggg ctcagacatc gtggcagatg ccgggaccat gccgcagctg 1140 cctttcgccg agaacttcgc catggcgcca agccacttcc gcccggagag catcgaagaa 1200 cagatgatgt tttccatggc tctttcttta gcagatggtc atggaagaac acactcgcaa 1260 gggttggcat ggttgtag 1278 <210> 2 <211> 1277 <212> DNA <213> Oryza sativa <400> 2 atggggaaca ggataggggg gaggaggaag gcgggggtgg aggagaggta cacgaggccg 60 caggggctgt acgagcacag ggacatcgac cagaagaagc tccggaagct gatcctcgag 120 gccaagctcg cgccgtgcta catgggcgcc gacgacgccg ccgccgccgc cgacctcgag 180 gagtgcccca tctgcttcct gtactaccca agtcttaacc gatcaaagtg ttgctcaaaa 240 gggatatgca ccgagtgctt tctccaaatg aaaccaactc acactgctca gcctacacaa 300 tgtccattct gcaaactccc agttatgctg tggagtatcg tggtgtaaag acaaaggagg 360 aaaggagcat agaacaattt gaagagcaga aagtcataga agcacaaatg aggatgcgcc 420 agcaagcact tcaagatgaa gaagataaga tgaaaagaaa acagaacagg tgctcttcta 480 gcagaacaat cacaccgacc aaagaagtgg agtatagaga tatttgcagc acatcctttt 540 cagtgccgtc ataccgatgt gctgagcaag aaactgaatg ctgttcatcg gaaccttcat 600 gctctgccca gactagcatg cgccctttcc attctaggca taaccgtgat gataacattg 660 acatgaatat agaggatatg atggttatgg aagcgatttg gcgttccatt caggagcagg 720 gaagtatagg gaatcctgtc tgtggcaact ttatgcctgt aactgagcca tctccgcgtg 780 aacgccagcc attcgttcca gctgcttctc tagaaatacc tcatggtggt ggattttcct 840 gtgcggttgc ggcaatggct gagcaccagc cacccagtat ggacttctct tacatggctg 900 gcagcagcgc attcccagtt ttcgacatgt tccggcgacc atgcaacatt gctggtggaa 960 gcatgtgtaa tctggagagc tcaccggaga gctggagcgg gatagcacca agctgcagca 1020 gggaagtggt aagagaagaa ggagagtgct cggctgacca ctggtcggag ggtgcagagg 1080 ccggaacaag ctacgcgggc tcagacatcg tggcagatgc cgggaccatg ccgcagctgc 1140 ctttcgccga gaacttcgcc atggcgccaa gccacttccg cccggagagc atcgaagaac 1200 agatgatgtt ttccatggct ctttctttag cagatggtca tggaagaaca cactcgcaag 1260 ggttggcatg gttgtag 1277

Claims (8)

A molecular marker composition for selecting a rice plant having increased zinc content compared to a wild type, comprising a GW2 gene in which a 316 base in the nucleotide sequence of SEQ ID NO: 1 is deleted. A rice plant screening primer having an increased zinc content compared to the wild type, comprising an oligonucleotide composed of 8 or more consecutive nucleotides comprising 315th base and 316th base in the nucleotide sequence of SEQ ID NO: 2 or a complementary oligonucleotide thereof set. A rice plant screening probe having an increased zinc content compared to the wild type, comprising an oligonucleotide composed of 8 or more consecutive nucleotides comprising 315th base and 316th base in the nucleotide sequence of SEQ ID NO: 2 or a complementary oligonucleotide thereof . A rice microorganism selecting microorganism having an increased zinc content compared to the wild type, comprising an oligonucleotide consisting of 8 or more consecutive nucleotides comprising 315th base and 316th base in the nucleotide sequence of SEQ ID NO: 2 or a complementary oligonucleotide thereof Array. A primer set of claim 2; And a reagent for carrying out an amplification reaction, wherein the zinc content is increased compared to the wild type. 6. The kit according to claim 5, wherein the reagent for carrying out the amplification reaction comprises a DNA polymerase, dNTPs and a buffer. Isolating the genomic DNA from the rice sample;
Amplifying the target sequence by using the separated genomic DNA as a template and carrying out an amplification reaction using the primer set of claim 2; And
And detecting the amplified product. A method for screening rice plants with increased zinc content compared to wild type. 8. The method of claim 7, wherein the detection of the amplification product is performed by capillary electrophoresis, DNA chip, gel electrophoresis, sequencing, radioactivity measurement, fluorescence measurement or phosphorescence measurement. A method for screening rice plants.

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