KR101546727B1 - Gene involved in leaf development from rice and uses thereof - Google Patents

Abstract

The present invention relates to a gene related to leaf development derived from rice and its use, and it is believed that the gene can contribute to enhancement of leaf production, growth and morphological characteristics of the plant, thereby improving crop yield and quality of production.

Description

[0002] Gene-related leaf development from rice and its uses [

[0001] The present invention relates to a gene relating to a leaf development derived from rice and a use thereof, and more particularly to a rice development-related protein derived from rice, a gene encoding the protein, a recombinant vector containing the gene, And a mutant plant having a phenotypic phenotype in comparison with the wild type, and a seed thereof.

In traditional cross-breed breeding, it takes a lot of time and effort to isolate and fix the traits, and because there are limited natural genetic resources that can be used, traditional cross-breed breeding is the only way to develop new varieties, It is difficult to go. Therefore, it is necessary to expand the basic research for the development of internationally competitive new varieties of rice using biotechnological techniques. To do this, it is important to find and secure genes that are useful for agriculture.

Leaf area and leaf angle improve photosynthetic ability, which is an important factor affecting plant growth and development.

Therefore, the present invention is to analyze and identify functions of useful genes related to such leaf development to secure agricultural useful genes.

Korean Laid-Open Patent Application No. 2013-0110926 discloses a new variety of grass in which the three-leaf mutation is induced, and Korean Patent Laid-Open Publication No. 2001-0068688 discloses a new variety of bermudagrass with berry characteristics. However, as described in the present invention, there is no description about a gene related to the phenotype of the rice leaf.

SUMMARY OF THE INVENTION The present invention has been made in view of the above-mentioned needs, and an object of the present invention is to provide a method of identifying knock-out genes from rice Ds insertion variants having a three- This is to isolate new useful genes related to leaf development, which is essential for the development of this increased crop, from rice and to identify its function.

In order to solve the above problems, the present invention provides a rice-related leaf development-related protein.

The present invention also provides a gene encoding the protein.

The present invention also provides a recombinant vector comprising the gene.

The present invention also provides a host cell transformed with the recombinant vector.

In addition, the present invention provides a mutant plant and a seed thereof, wherein the gene is knocked out and has a threefold phenotype as compared to the wild type.

According to the present invention, the morphological comparative analysis of the three-leaf mutants and the expression patterns in the rice plants were confirmed to be related to the leaf development genes. As a result, it is expected that the three - dimensional mutant can be used as a useful gene related to leaf development, growth and morphological development of the plant. This may be useful for other plants as well, and it may contribute to improve the yield and quality of crops by improving leaf development, growth and morphological characteristics of plants.

Figure 1 shows morphological characteristics of the wild type and the mutant. It was confirmed that the intersex length of the mutant was smaller than that of the wild type (A). The seeds ripening rate and maturation rate were lower in the ears than in the wild type (B). (D) The leaf type showed phenotype of expansion type compared to wild type (C), and the seed size was smaller and morphologically different (D).
Fig. 2 shows the nucleotide sequence of the knock-out gene of the three-dimensional mutant.
3 shows the amino acid sequence deduced from the cDNA of the knock-out gene of the three-leaf mutant.
FIG. 4 shows the result of analyzing the insertion position of T-DNA using a three-dimensional mutant. (A), and I-PCR was carried out for Flanking DNA analysis, and the result was confirmed by performing electrophoresis (B). As a result of observing the early growth of the wild type and the mutant, the mutant exhibited a phenotype of the leaf-like leaflet and an opening-type leaflet (C), and a homo system was selected using a knockout gene- (D).
FIG. 5 shows the result of confirming the expression of the knockout gene in the wild type rice by the parts in the panicle, leaf, sheath, internode and root of the plant .
Fig. 6 is a result of microscopic analysis of the regions A, B, and C of Dong Jinbyeong and mutants. The number of vascular bundles (LV) was small and the size was small in the mutant stem. In the number and size of starved cells (BC) and mesophyll cells in the vein, there was no difference between the two plants, but the mutants were small in overall size.

In order to accomplish the object of the present invention, the present invention provides a rice development-related protein derived from rice.

The range of the protein according to the present invention is rice ( Oryza comprising the amino acid sequence shown in the SEQ ID NO: 2 isolated from sativa) comprises the functional equivalent of a protein and the protein. Is at least 70% or more, preferably 80% or more, more preferably 90% or more, still more preferably 90% or more, more preferably 90% or more, Quot; refers to a protein having a homology of at least 95% with a physiological activity substantially equivalent to that of the protein represented by SEQ ID NO: 2. "Substantially homogenous physiological activity" means a three-dimensional phenotype. The invention also includes fragments, derivatives and analogues of the OsGASD protein. As used herein, the terms "fragments", "derivatives" and "analogs" refer to polypeptides having substantially the same biological function or activity as the OsGASD polypeptides of the present invention.

The present invention also provides a gene encoding the protein. The gene of the present invention relates to development of rice-derived leaves, and includes genomic DNA, cDNA, and synthetic DNA encoding the protein. Preferably, the gene of the present invention may include the nucleotide sequence shown in SEQ ID NO: 1. In addition, homologues of the nucleotide sequences are included within the scope of the present invention. Specifically, the gene includes a nucleotide sequence having a sequence homology of 70% or more, more preferably 80% or more, more preferably 90% or more, and most preferably 95% or more, with the nucleotide sequence of SEQ ID NO: 1 can do. "% Of sequence homology to polynucleotides" is ascertained by comparing the comparison region with two optimally aligned sequences, and a portion of the polynucleotide sequence in the comparison region is the reference sequence for the optimal alignment of the two sequences (I. E., A gap) relative to the < / RTI >

The present invention also provides a recombinant vector comprising the gene according to the present invention. In the recombinant vector of the present invention, the gene may preferably consist of the nucleotide sequence of SEQ ID NO: 1.

The term "recombinant" refers to a cell in which a cell replicates a heterologous nucleic acid, expresses the nucleic acid, or expresses a protein encoded by a peptide, heterologous peptide or heterologous nucleic acid. The recombinant cell can express a gene or a gene fragment that is not found in the natural form of the cell in one of the sense or antisense form. In addition, the recombinant cell can express a gene found in a cell in its natural state, but the gene has been modified and reintroduced intracellularly by an artificial means.

The term "vector" is used to refer to a DNA fragment (s), nucleic acid molecule, which is transferred into a cell. The vector replicates the DNA and can be independently regenerated in the host cell. The term "carrier" is often used interchangeably with "vector ". The term "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. In principle, any plasmid and vector can be used if it can replicate and stabilize within the host. An important characteristic of the expression vector is that it has a replication origin, a promoter, a marker gene and a translation control element. Enhancers, ribosome binding sites, termination signals, polyadenylation signals, and promoters, which are available in eukaryotic cells, are well known in the art. The expression vector can be constructed by a method known in the art. Such methods include in vitro recombinant DNA technology, DNA synthesis techniques, and in vivo recombination techniques. The DNA sequence can be effectively linked to appropriate promoters in the expression vector to drive mRNA synthesis. The expression vector may also include a ribosome binding site and a transcription terminator as a translation initiation site.

A preferred example of a recombinant vector is a Ti-plasmid vector which is capable of transferring a so-called T-region to a plant cell when it is present in a suitable host such as Agrobacterium tumefaciens. Other types of Ti-plasmid vectors (see EP 0 116 718 B1) are currently used to transfer hybrid DNA sequences to plant cells or protoplasts in which new plants capable of properly inserting hybrid DNA into the plant's genome can be produced have. A particularly preferred form of the Ti-plasmid vector is a so-called binary vector as claimed in EP 0 120 516 B1 and U.S. Patent No. 4,940,838. Other suitable vectors that can be used to introduce the DNA according to the invention into the plant host include viral vectors such as those that can be derived from double-stranded plant viruses (e. G., CaMV) and single- For example, from non -complete plant virus vectors. The use of such vectors may be particularly advantageous when it is difficult to transform the plant host properly.

In the recombinant vector of the present invention, the promoter may be a promoter suitable for transformation, preferably a CaMV 35S promoter, an actin promoter, a ubiquitin promoter, a pEMU promoter, a MAS promoter or a histone promoter, preferably a CaMV 35S promoter But is not limited thereto. The term "promoter " refers to the region of DNA upstream from the structural gene and refers to a DNA molecule to which an RNA polymerase binds to initiate transcription. A "plant promoter" is a promoter capable of initiating transcription in plant cells. A "constitutive promoter" is a promoter that is active under most environmental conditions and developmental conditions or cell differentiation. Since the selection of the transformant can be carried out by various tissues at various stages, a constant promoter may be preferable in the present invention. Therefore, the constant promoter does not limit the selectivity.

In the recombinant vector of the present invention, the terminator can be a conventional terminator, such as nopaline synthase (NOS), rice α-amylase RAmy1 A terminator, phaseoline terminator, Agrobacterium tumefaciens Agrobacterium tumefaciens , Octopine gene terminator of Agrobacterium tumefaciens , and the like. Regarding the need for a terminator, it is generally known that the terminator region increases the certainty and efficiency of gene transcription in plant cells. Therefore, the use of a terminator is highly desirable in the context of the present invention.

The recombinant vector of the present invention may preferably comprise one or more selectable markers. The marker is typically a nucleic acid sequence having a property that can be selected by a chemical method, and includes all genes capable of distinguishing a transformed cell from a non-transformed cell. Examples include herbicide resistance genes such as glyphosate or phosphinothricin, antibiotic resistant genes such as kanamycin, G418, Bleomycin, hygromycin, chloramphenicol, However, the present invention is not limited thereto.

Also provided is a host cell transformed with the recombinant vector of the present invention. Host cells capable of continuously cloning and expressing the vector of the present invention in a stable manner can be any host cell known in the art. Examples of prokaryotic cells include E. coli JM109, E. coli BL21, E. coli Bacillus sp. Strains such as RR1, E. coli LE392, E. coli B, E. coli X 1776, E. coli W3110, Bacillus subtilis, and Bacillus thuringiensis, and Salmonella typhimurium, Serratia marcesensis And enterobacteria and strains such as Pseudomonas spp.

When the vector of the present invention is transformed into eukaryotic cells, yeast ( Saccharomyce cerevisiae ), insect cells, human cells (e.g., Chinese hamster ovary, W138, BHK, COS-7, 293, HepG2 , 3T3, RIN and MDCK cell lines) and plant cells. The host cell is preferably a plant cell.

The recombinant vector of the present invention can be produced by using a CaCl ₂ method (Cohen, SN et al., Proc. Natl. Acac. Sci. USA, 9: 2110-2114 (Dower, WJ et al., Nucleic Acids Res., 16: 6127-6145 (1988)), etc., And can be delivered into cells. In addition, when the host cell is a eukaryotic cell, microinjection (Capecchi, MR, Cell, 22: 479 (1980)), calcium phosphate precipitation (Graham, FL et al., Virology, 52: 456 (Wong, TK et al., Gene, 10: 87 (1980)), DEAE- (Yang et al., Proc. Natl. Acad. Sci., 87: 9568-9572 (1990)) and dextran treatment (Gopal, Mol. Cell Biol., 5: 1188-1190 The vector can be injected into the host cell.

Further, the present invention provides a mutant plant and a seed thereof, wherein the gene is knocked out to have a three-leaf phenotype in comparison with the wild-type.

In one embodiment of the present invention, the gene may be a nucleotide sequence of SEQ ID NO: 1, but is not limited thereto.

In one embodiment of the present invention, the mutant plant is shorter in length than the wild type, and the shape of the leaflet shows the phenotype of the expansion type, and the ear has less maturity rate and maturity rate, The size of the seed is small, but the present invention is not limited thereto.

In one embodiment of the present invention, the plants used in the present invention are selected from the group consisting of Arabidopsis, potato, eggplant, cigarette, pepper, tomato, burdock, ciliaceae, lettuce, bellflower, spinach, modern sweet potato, celery, carrot, Such as rice, barley, wheat, rye, corn, sugar cane, oats, onions, etc., such as Chinese cabbage, cabbage, gangbu, watermelon, melon, cucumber, amber, pak, strawberry, soybean, mung bean, May be monocotyledons, preferably monocotyledons, more preferably Poaceae , and most preferably rice ( Oryza sativa , but is not limited thereto.

Hereinafter, the present invention will be described in detail with reference to examples. However, the following examples are illustrative of the present invention, and the present invention is not limited to the following examples.

Example  One. Ds Insertion mutant  Phenotypic analysis

In the present invention, a mutant having a three-dimensional phenotype among the Ds insertion mutants, which is a rice gene function analyzing material, was used. It was confirmed that the length of the intervertebral disc was smaller in the mutant than in the wild type (Fig. 1A). It was confirmed that seeds ripen and maturity were lower in the seeds than in the wild type (Fig. 1B). In the case of the leaf form, the phenotype of the expansion type was compared with that of the wild type (Fig. 1C), and the size of the seed was smaller and the morphological difference was observed (Fig. 1D).

Example  2. Isolation of genes related to leaf development derived from rice

Genomic DNA was isolated from the leaf of the present invention and the clean 1 copy homo transformant was selected by analyzing the insertion position of T-DNA through Flanking sequencing analysis. A cDNA sequence of the knockout gene was obtained using this. Based on this, a specific oligonucleotide primer was prepared and the gene was cloned using the PCR method. The knockout gene of the three-lobe mutant is a novel gene that has not yet been discovered. It has a total nucleotide sequence of 1140 bp and encodes 379 amino acids. The cDNA sequence and amino acid sequence of this gene are shown in detail in FIGS. 2 and 3, respectively.

The process of obtaining the gene information is as follows. Genomic DNA was isolated from the mutant and 2 ug of DNA was treated with NullII restriction enzyme treatment at 100 ° C for 2 hours and 30 minutes at 37 ° C, followed by treatment at 65 ° C for 30 minutes. 300 μl of 100% ethanol and 10 μl of 3M sodium acetate were added thereto, followed by treatment at -20 ° C. for 20 minutes, followed by centrifugation at 13000 rpm for 15 minutes to remove the supernatant. DNA was dissolved in 360 ul of distilled water, treated at 55 ° C for 10 minutes, self-ligated using T4-ligase, and cultured at 16 ° C for one day. DNA was then purified and purified by using primers Ds3-37 (5'-TATGAAAATGAAAACGGTTAGAGG-3 ') and Ds3I-105 (5'-AAACGAACGGGATAAATACGG-3' 15 sec, 55 [deg.] C for 1 min, 72 [deg.] C for 2 min and 30 sec) for 35 cycles and 72 [deg.] C for 10 min. The obtained PCR product was diluted to 1/100 and subjected to primary PCR using primers Ds3-4 (5'-GTTACCGACCGTTTTCATCC-3 ': SEQ ID NO: 5) and Ds3I-150 (5'-GGTTAAAGTCGAAATCGGACG- Secondary PCR was carried out in the same manner as in the above. The DNA obtained by electrophoresis of the first and second PCR products was purified and analyzed by Flanking sequence analysis (FIG. 4A, B). As a result of comparing the phenotype of the control and the three-leaf mutant, the mutant exhibited a smaller plant height and leaf length, a narrower leaf width, and a greater angle between the stem and leaf than the control (FIG. 4C). (5'-CACCATGGGCGCGGGGGCAGAGCA-3 ': SEQ ID NO: 7), reverse primer (5'-TCAGTTCTTGAGGTCAGGGA-3': SEQ ID NO: 8) and Ds3-4 primer to select a fixed system (Fig. 4D).

Example  3. Expression patterns of isolated genes

In order to confirm the expression pattern of the identified genes, the expression patterns of panicle, leaf, sheath, internode and root in Dongjinbio were examined. The samples were collected by Trizol reagent (Invitrogen), and qRT-PCR was performed by the following method. CDNA was prepared using the ReverTra Ace PCR RT Master mix (ToYoBo), and qRT-PCR was performed using SYBR (Takara). In this case, the gene specific forward primer 5'-CAATAAGGACTATAGCCGGTGG-3 '(SEQ ID NO: 9) and the reverse primer 5'-GGTCTACTCCACTAAAGCTCATG-3' (SEQ ID NO: 10) were used. As a result, the expression was most strongly expressed in the leaves and the expression level was low in other parts (FIG. 5).

Example  4. Cell-level Observation of morphological characteristics of mutants

Cellular level Microscopic analysis was performed to observe the morphological characteristics of the mutants. In order to analyze the morphological characteristics through the microscope, 0.5 cm of the middle part of the leaves of the wild type and the mutant was cut and placed in a fixing solution (formaldehyde 2.5%, acetic acid 2.5%, ethanol 45%, dH ₂ O 50%). Vacuum was used to maintain the vacuum for about 1 hour so that the fixer could penetrate well to the cut part of the plant. In order to remove the water content of the plant, ethanol treatment was performed at 50, 60, 70, 80, 90, 95, 99 and 100% (50 and 100% Samples treated with ethanol were incubated for 24 hours. Subsequently, the cells were replaced with 100% ethanol, and then the cells were substituted with Technovit solution (Technovit I 0.5 g + Technovit resion 50 ml) for 25 min, 50 min, 75 min and 100% The pretreatment was carried out at 4 ° C for 24 hours. Technovit block solution (Technovit Ⅱ 45μl + Technovit solution 750μl) was mixed well and poured into the mold. The pretreated plant parts were stained with netural red, and then put into the mold. It was hardened for 1 hour and 30 minutes, then placed in Technovit block solution on top and hardened. Thereafter, the block was sectioned to a thickness of 4 탆. After the section, the plant tissue was placed on a cover glass and the morphological characteristics were observed through a microscope. Observation of the morphological characteristics of the wild type and the single leaf mutant through the microscope showed that the leaf lobe mutant had narrower leaf width, smaller stem diameter, and fewer veins than the wild type . In order to compare the number and size of the cells, a comparative analysis was performed by dividing into a main vein (Fig. 6A), a large vein (Fig. 6B) and a small vein (Fig. 6C). In the number of vascular bundles in the main veins, the number of vascular bundles (LV) was smaller and the size was smaller than that of the wild type. In the number and size of starved cells (BC) and mesophyll cells in the vein, The mutant was small.

<110> Dong-A University Research Foundation For Industry-Academy Cooperation <120> Gene involved in leaf development from rice and uses thereof <130> PN13335 <160> 10 <170> Kopatentin 2.0 <210> 1 <211> 1140 <212> DNA <213> Oryza sativa <400> 1 atgggcgcgg aggcagagca ccgcatgtct ccctcccccg ctcccgctcc cgctccccct 60 acccctgttg ggaggaccga cggcgacgcc cctcccatgg tgctgggcct gcagctgtcg 120 gcgctcatcg accacgtcgc gcgcgtcgac tggtccctcc tcaaccgcat ccccggcgac 180 cgcggcggtt cacagcaggt ctgtattgag gagttgaatc atattctggc tgaagtgaat 240 gctcaaatcc tcccttgccg tgatgacctt tcttcaataa ggactatagc cggtggtagt 300 gtcgccaaca caattcgagg tctctcagca ggttttggga tatcaactgg aataattgga 360 gcttgtggag atgacagcca gggcgttttg tttgtcagca acatgagctt tagtggagta 420 gacctcacaa gattaaggac aaaaaagggg cacactgctc agtgtgcatg cttggtcgat 480 gcaagtggca atcgcaccat gcgcccttgc ctatctagtg cagtcaagct ccaggccaac 540 gagttcaaaa aggaggactt caaaggctcc aagtggttgg ttgtacgata tgcaagacag 600 aacatggaac agatccttga agctattagg attgctaaac aagaaggtct ctcagtatca 660 ttggatttgg ctagttttga gatggttcgt gactatagga cacaactcat tgaccttttg 720 gaaactggca acattgacct ttgctttgcc aatgaggatg aagctagaga gcttctaggg 780 ggagagctga catttgatcc agaggaggca cttgctttct tggccaagta ctgtaaatgg 840 gccgtggtga ccctagcttc taagggatgc atagcgaagc atgggaagca ggtggttcaa 900 gtggcggcaa ccggggagag taatgcggtg gacgcgacgg gggcggggga tctgtttgcg 960 agcgggttcc tgtacgggtt ggtgaaaggg ctggcattgg aggagtgctg caaggtaggg 1020 gcgtgcagcg gtgggtcagt ggtgagggct ctgggcgggg aggtgaggcc ggagaactgg 1080 cagtggatgt acaagcagat gaacgccagt ggcctgctgc tccctgacct caagaactga 1140                                                                         1140 <210> 2 <211> 379 <212> PRT <213> Oryza sativa <400> 2 Met Gly Ala Glu Ala Glu His Arg Met Ser Ser Pro Ala Pro Ala   1 5 10 15 Pro Ala Pro Pro Thr Pro Val Gly Arg Thr Asp Gly Asp Ala Pro Pro              20 25 30 Met Val Leu Gly Leu Gln Leu Ser Ala Leu Ile Asp His Val Ala Arg          35 40 45 Val Asp Trp Ser Leu Leu Asn Arg Ile Pro Gly Asp Arg Gly Gly Ser      50 55 60 Gln Gln Val Cys Ile Glu Glu Leu Asn His Ile Leu Ala Glu Val Asn  65 70 75 80 Ala Gln Ile Leu Pro Cys Arg Asp Asp Leu Ser Ser Ile Arg Thr Ile                  85 90 95 Ala Gly Gly Ser Val Ala Asn Thr Ile Arg Gly Leu Ser Ala Gly Phe             100 105 110 Gly Ile Ser Thr Gly Ile Ile Gly Ala Cys Gly Asp Asp Ser Gln Gly         115 120 125 Val Leu Phe Val Ser Asn Met Ser Phe Ser Gly Val Asp Leu Thr Arg     130 135 140 Leu Arg Thr Lys Lys Gly His Thr Ala Gln Cys Ala Cys Leu Val Asp 145 150 155 160 Ala Ser Gly Asn Arg Thr Met Arg Pro Cys Leu Ser Ser Ala Val Lys                 165 170 175 Leu Gln Ala Asn Glu Phe Lys Lys Glu Asp Phe Lys Gly Ser Lys Trp             180 185 190 Leu Val Val Arg Tyr Ala Arg Gln Asn Met Glu Gln Ile Leu Glu Ala         195 200 205 Ile Arg Ile Ala Lys Gln Glu Gly Leu Ser Val Ser Leu Asp Leu Ala     210 215 220 Ser Phe Glu Met Val Arg Asp Tyr Arg Thr Gln Leu Ile Asp Leu Leu 225 230 235 240 Glu Thr Gly Asn Ile Asp Leu Cys Phe Ala Asn Glu Asp Glu Ala Arg                 245 250 255 Glu Leu Leu Gly Gly Glu Leu Thr Phe Asp Pro Glu Glu Ala Leu Ala             260 265 270 Phe Leu Ala Lys Tyr Cys Lys Trp Ala Val Val Thr Leu Ala Ser Lys         275 280 285 Gly Cys Ile Ala Lys His Gly Lys Gln Val Val Gln Val Ala Ala Thr     290 295 300 Gly Glu Ser Asn Ala Val Asp Ala Thr Gly Ala Gly Asp Leu Phe Ala 305 310 315 320 Ser Gly Phe Leu Tyr Gly Leu Val Lys Gly Leu Ala Leu Glu Glu Cys                 325 330 335 Cys Lys Val Gly Ala Cys Ser Gly Gly Ser Val Val Arg Ala Leu Gly             340 345 350 Gly Glu Val Arg Pro Glu Asn Trp Gln Trp Met Tyr Lys Gln Met Asn         355 360 365 Ala Ser Gly Leu Leu Leu Pro Asp Leu Lys Asn     370 375 <210> 3 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 3 tatgaaaatg aaaacggtag agg 23 <210> 4 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 4 aaacgaacgg gataaatacg g 21 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 5 gttaccgacc gttttcatcc 20 <210> 6 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 6 ggttaaagtc gaaatcggac g 21 <210> 7 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 7 caccatgggc gcggggcaga gca 23 <210> 8 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 8 tcagttcttg aggtcaggga 20 <210> 9 <211> 22 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 9 caataaggac tatagccggt gg 22 <210> 10 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Primer <400> 10 ggtctactcc actaaagctc atg 23

Claims (10)

delete delete delete delete delete A mutant plant comprising an amino acid sequence of SEQ ID NO: 2, wherein a gene encoding a rice-derived leaf development-related protein is knocked out and has a three-dimensional phenotype as compared to a wild-type. [Claim 7] The plant according to claim 6, wherein the plant is shorter in length than the wild type, the shape of the leaf is in the developmental phenotype, the seeds have less maturity and maturation rate, fewer veins, A mutant plant characterized by small. The mutant plant according to claim 6, wherein the plant is a Poaceae plant. 7. The mutant plant according to claim 6, wherein the gene consists of the nucleotide sequence of SEQ ID NO: 1. A seed of a mutant plant according to claim 6.

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