KR101156017B1 - Cytochrome P450 gene of giant embryonic rice and rice plant comprising the same - Google Patents

Abstract

The present invention relates to a rice plant having a giant pear, which encodes the amino acid sequence of cytochrome P450, and has a cytochrome P450 gene of rice having a giant pear comprising a nucleotide sequence encoding leucine as the 395th amino acid after translation, and the same. It provides a rice plant having.

Description

Cytochrome P450 gene of giant embryonic rice and rice plant comprising the same}

The present invention relates to a rice plant having a giant pear, and more particularly, the weight ratio of the seed of brown rice is heavier than the existing giant pear varieties, and also contains a lot of inorganic components such as zinc and iron to provide rice with enhanced nutrition. The present invention relates to a cytochrome P450 gene of rice and a rice plant containing the same, which has a huge pear capable of creating new demand for rice and enhancing competitiveness of the rice industry.

To date, giant cultivar rice varieties have been developed mainly by treating seeds with MNU (Hong et al., 2006; Hirosh et al., 2001), which is a mutant to rice seeds, or gamma rays (Kageyama et al., 2001), a kind of radiation. Giant embryo genes are located on chromosome 7 of rice (Nagasawa, 2002; Koh, 1996) and are known to be caused by mutations in cytochrome P450 gene specific sequences (Cahoon et al., US Patent No Us 2003/0126645 A1). .

However, the huge pear rice varieties known to date have a disadvantage that the size of the pear is not large enough, and it is somewhat inferior in cooking characteristics and sensuality, and also inferior in terms of nutritional value due to insufficient inorganic ingredients such as zinc and iron. have.

In the present invention, the weight ratio of the seed grain in brown rice is heavier than the existing giant pear varieties, and also contains abundant inorganic components such as zinc and iron, which can provide rice with enhanced nutrition, thereby creating new demand for rice and competitiveness of the rice industry. The present invention provides a cytochrome P450 gene of rice having a giant pear and a rice plant containing the same.

The technical problem as described above is achieved by the following configuration according to the present invention.

(1) A cytochrome P450 gene of rice having a giant embryo, which encodes an amino acid sequence of cytochrome P450 and which contains a nucleotide sequence encoding leucine as the 395th amino acid after translation.

(2) The cytochrome P450 gene according to item 1, wherein the gene consisting of the nucleotide sequence is a cDNA sequence encoding cytochrome P450, of which the 1195th base is T.

(3) The cytochrome P450 gene according to the above 1, wherein the gene consisting of the nucleotide sequence is a genomic gene represented by SEQ ID NO: 1.

(4) The cytochrome P450 gene according to the above 1, wherein the gene consisting of the nucleotide sequence is a cDNA gene represented by SEQ ID NO: 2.

(5) The cytochrome P450 gene according to the above 1, wherein the nucleotide sequence comprises a gene encoding an amino acid sequence of cytochrome P450 represented by SEQ ID NO: 3.

(6) A recombinant vector containing the gene according to 1 above.

(7) Rice plants having a giant embryo transformed with the recombinant vector according to the above 6.

(8) A rice plant having a giant embryo having a cytochrome P450 gene which codes for the amino acid sequence of cytochrome P450 and which contains a nucleotide sequence encoding leucine as the 395th amino acid after translation.

(9) The huge pear rice plant according to the above 8, wherein the rice having a huge pear is YR23517Acp79, which is a rice plant.

According to the above configuration of the present invention, the weight ratio of the seeds in brown rice is heavier than the existing giant pear varieties, and also contains a lot of inorganic components such as zinc and iron, which can provide rice with enhanced nutrition, thereby creating new demand for rice. This will enhance the competitiveness of the rice industry.

Hereinafter, the content of the present invention will be described in detail with reference to the drawings.

The present invention encodes the amino acid sequence of cytochrome P450 of rice, and includes the cytochrome P450 gene of rice having a huge embryo comprising a nucleotide sequence encoding leucine to the 395th amino acid after translation.

The gene according to the present invention may be a gene represented by SEQ ID NO: 1. The gene represented by SEQ ID NO: 1 was identified as a specific allele of genomic DNA isolated from the giant embryonic rice plant according to the present invention, and found to include a gene encoding cytochrome P450 as a result of homology analysis of the nucleotide sequence.

The gene represented by SEQ ID NO: 1 includes additional sequence information upstream and downstream as well as an open reading frame (ORF), and includes an intron in the open reading frame.

The gene of SEQ ID NO: 1 can be obtained by cloning method from genomic DNA of giant embryonic rice plant "YR23517Acp79" included in the present invention. For example, after primers are appropriately synthesized using the nucleotide sequence of SEQ ID NO: 1, To perform the PCR method.

The present invention is a nucleotide sequence of ge ^t gene (cDNA) contained in the genome nucleotide sequence of SEQ ID NO: 1 shown in SEQ ID NO: 2. According to a database search result, the gene of SEQ ID NO: 2 is located at Os07g0603700 (http://www.ncbi.nlm.nih.gov/) and LOC_Os07g41240 (http://www.gramene.org/). According to homology studies, the gene of SEQ ID NO: 2 shows 99.92% and 99% identity to cytochrome P450 found in plants and nucleotide and amino acid sequences, respectively. Therefore, the gene of the present invention is judged to be a novel allele belonging to cytochrome P450.

The 1184th base of the ge ^t gene according to the present invention is characterized in that T. This is not found in other cytochrome P450 genes known to date. This variation in base results in the transformation of the 395th amino acid of cytochrome P450 from tryptophan (W) to leucine (L).

The gene of SEQ ID NO: 2 can be obtained by RT-PCR and cloning method from RNA extracted from embryos of the embryonic seed of the giant embryonic rice plant "YR23517Acp79" included in the present invention. The primer is appropriately synthesized using the sequence, followed by conventional PCR or RT-PCR methods.

In addition, the present invention is the amino acid sequence of cytochrome P450 encoded by the cDNA of SEQ ID NO: 2 represented by SEQ ID NO: 3. The open reading frame (ORF) of SEQ ID NO: 3 is composed of 520 amino acids, and the mutation of the 395th amino acid of cytochrome P450 is changed from tryptophan (W) to leucine (L). Becomes a huge ship. This was found to be an allele different from the cytochrome P450 amino acid sequence of the giant embryonic mutant known to date.

The ge ^t gene according to the present invention as described above can be used as a practical selection marker for future breeding of giant pear rice and the giant pear rice plant can be used as a material for producing products using not only grain but also seed and seed-containing components of rice. have.

Giant pear rice plant according to the present invention can be selected by the following process. first

1) Rice Varieties Cultivated hybrid plants (YR23517) were artificially bred using Hwacheong Paddy as a model and Sangju Sticky Paddy as a copy.

2) Then, the ear which shows pollen development status of the early and the middle stage of the first nuclear stage is collected, and then the ear is collected by carrying out the cultivation according to the conventional method.

3) The collected ear is soaked in N ₆ -Y ₂ medium containing 2ppm NAA, 0.2ppm cinetin, etc., and then low-temperature treatment for 15 days, followed by incubation at 25 ° C for 30-40 days to induce callus.

4) The transferred callus is transferred to N ₆ -Y ₂ medium containing 0.2 ppm IAA, 2 ppm cinetin, etc., and cultured for about 30 days to regenerate the plant.

5) Plants are transplanted into culture bottles to induce root development and transfer to the soil to harvest seeds of each plant.

6) The harvested seeds are transferred back to the soil, and the grain characteristics of each plant can be analyzed to select a giant pear-rice plant.

Giant pear rice plant according to the present invention has several tillers in a single bar, can be asexual propagation through the method of artificially separating and re-planting the grains of the giant pear rice plant, and breeding using seeds Proliferation is also possible without a change in properties.

In the present invention, the above-mentioned rice paddy plant is described by selecting from plants obtained by artificial breeding between plants, but it is also possible to manufacture by transforming a plant using a genetic recombinant invention.

Methods that can be adopted for the transformation of rice do not require special limitations, and include, for example, microprojection bombardment of in vitro cultures or suspension cells and direct gene uptake or electroporation of protoplasts. .

Recombinant vectors that can be used for the production of such transformants are sufficient to carry out according to known commercial methods. Regulatory sequences already known to express these recombinant genes at high or appropriate levels can also be used in the present invention. Such regulatory sequences may be obtained from plants or plant viruses or synthesized by chemical methods. Such regulatory sequences may include promoters that direct transcription. Examples of promoters for increasing the expression rate include, but are not limited to, promoters of storage protein genes such as the CruA promoter of Brassica napus (Ryan et al., 1989) or promoters of structural expression genes such as the 35S promoter of CaMV (Cauliflower Mosaic Virus). (Guilley et al., 1982). Other regulatory sequences may include termination sequences and polyadenylation signals and all sequences that function in plants. Specific examples include the 3 'locus of the nopaline synthase gene of Agrobacterium tumafaciene or the 3' locus of the CurA gene of Brassica napus. Regulatory sequences may also include enhancer sequences such as those found in the 35S promoter of CaMV, mRNA stabilization sequences (Brederode et al., 1980), or other sequences that function such as AIMV (Alfalfa Mosaic Virus) RNA4 leader sequences. Can be.

All parts (promoter, regulatory sequence, stabilization sequence, signal sequence or termination sequence) of suitable gene constructs of the present invention can be mutated to change their regulatory properties using known techniques, if necessary.

Hereinafter, the content of the present invention will be described in more detail with reference to Examples. However, the following illustrated examples are only presented to aid the understanding of the present invention and should not be construed as limiting the scope of the present invention.

Example 1 Selection of Giant Rice YR23517Acp79 by Medicine Culture

Rice varieties Hwacheong rice and Sangjuchal rice are varieties with normal sized pears. The hybrid plants (YR23517), which were artificially crossed, were grown using Hwachung rice as a model and Sangjuchal rice as a copy. Ears showing pollen development in the early and mid-nuclear stage were harvested and then cultured according to the conventional method. The collected ear was soaked in N ₆ -Y ₂ medium containing 2ppm NAA, 0.2ppm cinetin, and the like for 15 days, and then incubated at 25 ° C. for 30-40 days to induce callus.

The callus was transferred to N ₆ -Y ₂ medium containing 0.2ppm IAA, 2ppm cinetin and the like, and cultured for about 30 days to re-differentiate the plant. Plants were transplanted into culture bottles to induce root development, seeded in soil and harvested seeds of each plant. As a result of analyzing the grain characteristics of each plant by transferring the harvested seeds back to the soil, we selected the YR23517Acp79, a giant pear rice plant with large eyes, and sent it to the Agricultural Genetic Resource Center, National Agricultural Science Institute, Seodun-dong, Suwon-si, Gyeonggi-do. Deposited with accession number KACC 98006P as of date.

The grain size of the giant pear rice plant "YR23517Acp79" is significantly larger than that of ordinary rice (Fig. 1B), as shown in Figs. 1A and 1C. In addition, `` YR23517Acp79 '' had a weight ratio of 16.5% of brown rice seeds, which was heavier than the general giant pear `` big eyes '' and contained more inorganic components such as zinc and iron than `` big eyes '' (Table 1).

<Table 1> Composition and composition of giant pear rice plant "YR23517Acp79" and general giant pear variety "big eyes"

division Lip count Seaweed weight (A) Oil drainage weight (B) A / B (%) Inorganic ingredients Remarks Zn Fe YR23517Acp79 400 0.758 g 4.597 g 16.5 42 ppm 33 ppm Rice Big eyes 400 0.558 g 6.209 g 9.0 27 ppm 24 ppm Buckwheat

Example 2: Cloning and sequencing of ge ^t genes

Cytochrome P450 (LOC_Os07g41240) genome, which is a gene related to seed size, from the giant cultivar rice plant `` YR23517Acp79 '', which was selected from the later generations of the cultivated rice varieties of hwacheong and sangju rice in the present invention. Gene was isolated.

Using DNA cytochrome P450 genome sequence obtained through NCBI database (http://www.ncbi.nlm.nih.gov/) for gene isolation by PCR, each DNA was divided into two DNA fragments around introns. Primers were synthesized appropriately according to the nucleotide sequence of the fragments, and the conventional PCR method was performed under the condition that 5 μl of betaine (5 μg) was added (sigma, USA). PCR reaction conditions were 50 μl of a mixture containing 20 ng of DNA, 500 mM betaine, 1 × Ex Taq buffer, 0.2 mM dNTP, 0.5 unit Ex Taq polymerase (Takara, Japan), and 1 uM of primers. Amplified times. Primers used in the PCR are as follows.

SEQ ID NO: P450F1 5'-TAGCTTCTCTCCACGTCTT-3 '(forward primer)

SEQ ID NO: P450R1 5'-GCCATGGACGTTTCCTTCTC-3 '(reverse primer)

SEQ ID NO: P450F2 5'-TCTCCTCGTCGTCGTCTTC-3 '(forward primer)

SEQ ID NO: P450R2 5'-TGATGTTTCACAACTAATGCCTCG-3 '(reverse primer)

Two genomic DNAs amplified with the four primers were conjugated to pGEM-T Easy Vector (Promega Co, USA) and transformed into recombinant E. coli JM109 cells. The transformed E. coli with the recombinant vector was first selected through the color selection (blue) using antibiotic resistance markers (Epicillin) and beta galactosidase enzyme, and the selected E. coli was inserted through DNA miniprep process. The vector was extracted and confirmed secondary. Through such a series of processes, a transgenic E. coli with the genomic sequence of each cytochrome P450 gene was inserted.

The nucleotide sequence of the cytochrome P450 gene inserted into the vector was identified by the sequencing primers T7 (SEQ ID NO: 5: 5'-TAATACGACTCACTATAGGG-3 ') and SP6 (SEQ ID NO: 6: 5'-ATTTAGGTGACACTATAG-3'). The nucleotide sequence of cytochrome P450 genomic DNA was analyzed. The sequencing work was confirmed to be a very accurate result obtained by performing two sequencing of each of the 10 recombinant DNAs obtained independently to eliminate sequencing errors.

The intron portion of the genomic DNA obtained through the sequencing analysis was removed, and for the gene and the amino acid sequence consisting of exons (cDNA), sangcheong rice (the model), sangjujub rice (the copy), and the nibbarae ( The homology with cytochrome P450 found in Nipponbare) and Dongjin rice was investigated.

As shown in FIG. 2, the results of the sequencing analysis showed that the 1195th base in the cytochrome P450 genomic DNA of the nibbarae, hwacheong rice, sangju rice, and dongjin rice had the normal size. As the (thymine) it was confirmed that the 395th amino acid is modified from tryptophan (W) to leucine (L). Therefore, this gene (cDNA) was named ge ^t , a new allele of the cytochrome P450 gene (cDNA). The rice with the ge ^t gene is a giant embryo with an increased size of seed.

While the present invention has been particularly shown and described with reference to exemplary embodiments thereof, it will be understood by those skilled in the art that various changes and modifications may be made without departing from the spirit and scope of the invention as defined in the appended claims. It can be understood that

1 is a comparative photograph of a grain of a giant pear rice plant "YR23517Acp79" according to the present invention and a grain of ordinary rice.

Figure 2 is a comparative sequence showing the mutated site of the gene of the giant pear rice plant "YR23517Acp79" according to the present invention compared with that of other plants.

<110> REPUBLIC OF KOREA (MANAGEMENT: RURAL DEVELOPMENT ADMINISTRATION) <120> Cytochrome P450 gene of giant embryonic rice and rice plant          configuring the same <160> 7 <170> Kopatentin 1.71 <210> 1 <211> 2301 <212> DNA <213> Rice plant <220> <221> gene (222) (1) .. (2301) <223> genomic DNA of cytochrome P450 <400> 1 gccattatta tgtagtccta tatttaagga agaaactaat gatatatacg cagatattgt 60 taataatgac cctttgatta cgctatcatt actgacaatg acatgtgggg ctagagtgtc 120 agataattgg aggtccaaat ttttggagtg gcaaaatggt ctatttaaag caccaggtgt 180 ttattagctt ctctccacgt cttcttcctc ccaagaaaac tcctctcact tcgcgaacgc 240 ttcccatggc gctctcctcc atggccgcgg cgcaagagag ctccctcctc ctcttcctcc 300 tcccgacgtc ggccgcctcc gtgttcccgc cgctcatctc cgtggtcgtc ctcgccgcgc 360 tcctcctgtg gctctcgccg ggtggccccg cgtgggcgct gtcccgttgc cgtggcacgc 420 cgccgccgcc gggcgtggcg gggggcgcgg ccagcgcgct gtccggccct gccgcgcacc 480 gcgtgctcgc cgggatttcg cgcgccgtcg agggcggcgc ggcggtgatg tcgctctccg 540 tcggcctcac ccgcctcgtc gtggcgagcc ggccggagac ggcgagggag atcctcgtca 600 gcccggcgtt cggcgaccgc cccgtgaagg acgcggcgag gcagctgctg ttccaccgcg 660 ccatggggtt cgccccgtcg ggcgacgcgc actggcgcgg gctccgccgc gcctccgcgg 720 cgcacctctt cggcccgcgc cgcgtggccg ggtccgcgcc cgagcgcgag gccatcggcg 780 cccgcatagt cggcgacgtc gcctccctca tgtcccgccg cggcgaggtc cccctccgcc 840 gcgtccttca cgccgcgtcg ctcggccacg tcatggcgac cgtcttcggc aagcggcacg 900 gcgacatctc gatccaggac ggcgagctcc tggaggagat ggtcaccgaa gggtacgacc 960 tcctcggcaa gttcaactgg gccgaccacc tgccattgct caggtggctc gacctccagg 1020 gcatccgccg ccggtgcaac aggctagtcc agaaggtgga ggtgttcgtc ggaaagatca 1080 tacaggagca caaggcgaag cgagctgccg gaggcgtcgc cgtcgccgac ggcgtcttgg 1140 gcgacttcgt cgacgtcctc ctcgacctcc agggagagga gaagatgtca gactccgaca 1200 tgatcgctgt tctttgggta agtctcctcg tcgtcgtctt cgtcgtaaag cttgagaagg 1260 aaacgtccat ggcgttttca tggattggtt tcttgttttt ttcttcagga gatgatcttt 1320 agagggacgg acacggtggc gatcttgatg gagtgggtga tggcgaggat ggtgatgcac 1380 ccggagatcc aggcgaaggc gcaggcggag gtggacgccg ccgtgggggg acgccgcggc 1440 cgcgtcgccg acggcgacgt ggcgagcctc ccctacatcc agtccatcgt gaaggagacg 1500 ctgcgcatgc acccgccggg cccgctcctg tcgtgggcgc gcctcgccgt gcacgacgcg 1560 cgcgtcggtg gccacgccgt ccccgccggg acgacggcga tggtgaacat gtgggcgatc 1620 gcccacgacg ccgccgtctg gccggagccg gatgcgttcc gcccggagcg cttctcggag 1680 ggggaggacg tcggcgtgct cggcggcgac ctccgcctcg cgccgttcgg cgccggccgc 1740 cgcgtctgcc ctggcaggat gctggcgctc gccaccgccc acctctggct cgcccagctg 1800 ctgcacgcct tcgactggtc ccccaccgcc gccggcgtcg acctgtccga gcgcctcggc 1860 atgtcgctgg agatggcggc gccgctcgtg tgcaaggccg tggctagggc ctgagcccta 1920 gccgccgccg ccgccattat tgccattgat gtggctagcg acgttgtcgt gctcgcatcc 1980 atactcctcc ataggcaact cgtctagcca atgaagaaag ctactatcta tctatctatc 2040 aagctagctg ctactatcac aaaccgcatt tcggcatcat cttaaattag ctcttagggg 2100 tgtaggcgat tttggtttcc cccaaaaatt tgctttgcca gtcttttggt ttaaatcgag 2160 gcattagttg tgaaacatca tgagaagtta tttaaatctg aggaattttg tttgaacctt 2220 ttctggtgtg ctaaatggat cgtgctttga gtatcttatt attctgaatg tgttatgtag 2280 ctacactctc ctgaatcatg t 2301 <210> 2 <211> 1563 <212> DNA <213> Rice plant <220> <221> gene (222) (1) .. (1560) <223> cDNA of cytochrome P450 <400> 2 atggccgcgg cgcaagagag ctccctcctc ctcttcctcc tcccgacgtc ggccgcctcc 60 gtgttcccgc cgctcatctc cgtggtcgtc ctcgccgcgc tcctcctgtg gctctcgccg 120 ggtggccccg cgtgggcgct gtcccgttgc cgtggcacgc cgccgccgcc gggcgtggcg 180 gggggcgcgg ccagcgcgct gtccggccct gccgcgcacc gcgtgctcgc cgggatttcg 240 cgcgccgtcg agggcggcgc ggcggtgatg tcgctctccg tcggcctcac ccgcctcgtc 300 gtggcgagcc ggccggagac ggcgagggag atcctcgtca gcccggcgtt cggcgaccgc 360 cccgtgaagg acgcggcgag gcagctgctg ttccaccgcg ccatggggtt cgccccgtcg 420 ggcgacgcgc actggcgcgg gctccgccgc gcctccgcgg cgcacctctt cggcccgcgc 480 cgcgtggccg ggtccgcgcc cgagcgcgag gccatcggcg cccgcatagt cggcgacgtc 540 gcctccctca tgtcccgccg cggcgaggtc cccctccgcc gcgtccttca cgccgcgtcg 600 ctcggccacg tcatggcgac cgtcttcggc aagcggcacg gcgacatctc gatccaggac 660 ggcgagctcc tggaggagat ggtcaccgaa gggtacgacc tcctcggcaa gttcaactgg 720 gccgaccacc tgccattgct caggtggctc gacctccagg gcatccgccg ccggtgcaac 780 aggctagtcc agaaggtgga ggtgttcgtc ggaaagatca tacaggagca caaggcgaag 840 cgagctgccg gaggcgtcgc cgtcgccgac ggcgtcttgg gcgacttcgt cgacgtcctc 900 ctcgacctcc agggagagga gaagatgtca gactccgaca tgatcgctgt tctttgggag 960 atgatcttta gagggacgga cacggtggcg atcttgatgg agtgggtgat ggcgaggatg 1020 gtgatgcacc cggagatcca ggcgaaggcg caggcggagg tggacgccgc cgtgggggga 1080 cgccgcggcc gcgtcgccga cggcgacgtg gcgagcctcc cctacatcca gtccatcgtg 1140 aaggagacgc tgcgcatgca cccgccgggc ccgctcctgt cgttggcgcg cctcgccgtg 1200 cacgacgcgc gcgtcggtgg ccacgccgtc cccgccggga cgacggcgat ggtgaacatg 1260 tgggcgatcg cccacgacgc cgccgtctgg ccggagccgg atgcgttccg cccggagcgc 1320 ttctcggagg gggaggacgt cggcgtgctc ggcggcgacc tccgcctcgc gccgttcggc 1380 gccggccgcc gcgtctgccc tggcaggatg ctggcgctcg ccaccgccca cctctggctc 1440 gcccagctgc tgcacgcctt cgactggtcc cccaccgccg ccggcgtcga cctgtccgag 1500 cgcctcggca tgtcgctgga gatggcggcg ccgctcgtgt gcaaggccgt ggctagggcc 1560 tga 1563 <210> 3 <211> 520 <212> PRT <213> Rice plant <220> <221> PEPTIDE (222) (1) .. (520) <223> Cytochrome P450 <400> 3 Met Ala Ala Ala Gln Glu Ser Ser Leu Leu Leu Phe Leu Leu Pro Thr   1 5 10 15 Ser Ala Ala Ser Val Phe Pro Pro Leu Ile Ser Val Val Val Leu Ala              20 25 30 Ala Leu Leu Leu Trp Leu Ser Pro Gly Gly Pro Ala Trp Ala Leu Ser          35 40 45 Arg Cys Arg Gly Thr Pro Pro Pro Pro Gly Val Ala Gly Gly Ala Ala      50 55 60 Ser Ala Leu Ser Gly Pro Ala Ala His Arg Val Leu Ala Gly Ile Ser  65 70 75 80 Arg Ala Val Glu Gly Gly Ala Ala Val Met Ser Leu Ser Val Gly Leu                  85 90 95 Thr Arg Leu Val Val Ala Ser Arg Pro Glu Thr Ala Arg Glu Ile Leu             100 105 110 Val Ser Pro Ala Phe Gly Asp Arg Pro Val Lys Asp Ala Ala Arg Gln         115 120 125 Leu Leu Phe His Arg Ala Met Gly Phe Ala Pro Ser Gly Asp Ala His     130 135 140 Trp Arg Gly Leu Arg Arg Ala Ser Ala Ala His Leu Phe Gly Pro Arg 145 150 155 160 Arg Val Ala Gly Ser Ala Pro Glu Arg Glu Ala Ile Gly Ala Arg Ile                 165 170 175 Val Gly Asp Val Ala Ser Leu Met Ser Arg Arg Gly Glu Val Pro Leu             180 185 190 Arg Arg Val Leu His Ala Ala Ser Leu Gly His Val Met Ala Thr Val         195 200 205 Phe Gly Lys Arg His Gly Asp Ile Ser Ile Gln Asp Gly Glu Leu Leu     210 215 220 Glu Glu Met Val Thr Glu Gly Tyr Asp Leu Leu Gly Lys Phe Asn Trp 225 230 235 240 Ala Asp His Leu Pro Leu Leu Arg Trp Leu Asp Leu Gln Gly Ile Arg                 245 250 255 Arg Arg Cys Asn Arg Leu Val Gln Lys Val Glu Val Phe Val Gly Lys             260 265 270 Ile Ile Gln Glu His Lys Ala Lys Arg Ala Ala Gly Val Ala Val         275 280 285 Ala Asp Gly Val Leu Gly Asp Phe Val Asp Val Leu Leu Asp Leu Gln     290 295 300 Gly Glu Glu Lys Met Ser Asp Ser Asp Met Ile Ala Val Leu Trp Glu 305 310 315 320 Met Ile Phe Arg Gly Thr Asp Thr Val Ala Ile Leu Met Glu Trp Val                 325 330 335 Met Ala Arg Met Val Met His Pro Glu Ile Gln Ala Lys Ala Gln Ala             340 345 350 Glu Val Asp Ala Ala Val Gly Gly Arg Arg Gly Arg Val Ala Asp Gly         355 360 365 Asp Val Ala Ser Leu Pro Tyr Ile Gln Ser Ile Val Lys Glu Thr Leu     370 375 380 Arg Met His Pro Pro Gly Pro Leu Leu Ser Leu Ala Arg Leu Ala Val 385 390 395 400 His Asp Ala Arg Val Gly Gly His Ala Val Pro Ala Gly Thr Thr Ala                 405 410 415 Met Val Asn Met Trp Ala Ile Ala His Asp Ala Ala Val Trp Pro Glu             420 425 430 Pro Asp Ala Phe Arg Pro Glu Arg Phe Ser Glu Gly Glu Asp Val Gly         435 440 445 Val Leu Gly Gly Asp Leu Arg Leu Ala Pro Phe Gly Ala Gly Arg Arg     450 455 460 Val Cys Pro Gly Arg Met Leu Ala Leu Ala Thr Ala His Leu Trp Leu 465 470 475 480 Ala Gln Leu Leu His Ala Phe Asp Trp Ser Pro Thr Ala Ala Gly Val                 485 490 495 Asp Leu Ser Glu Arg Leu Gly Met Ser Leu Glu Met Ala Ala Pro Leu             500 505 510 Val Cys Lys Ala Val Ala Arg Ala         515 520 <210> 4 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> forward primer P450F1 <400> 4 tagcttctct ccacgtctt 19 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> reverse primer of P450R1 <400> 5 gccatggacg tttccttctc 20 <210> 6 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> forward primer of P450F2 <400> 6 tctcctcgtc gtcgtcttc 19 <210> 7 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> reverse primer of P450R2 <400> 7 tgatgtttca caactaatgc ctcg 24  

Claims (9)

A cytochrome P450 gene of rice represented by SEQ ID NO: 2 and having a huge fold consisting of a nucleotide sequence encoding leucine with a 395th amino acid after translation of the 1184th base into T. Recombinant vector containing the gene according to claim 1. Rice plant having a giant embryo transformed with the recombinant vector according to claim 2. 4. The rice plant according to claim 3, wherein the rice having a giant pear is YR23517Acp79 (KACC 98006P), which is a waxy plant. delete delete delete delete delete

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