CN109371000B - Rice ACCase mutant type gene and application thereof in herbicide resistance of plants - Google Patents

Abstract

The invention discloses a rice ACCase mutant gene and application thereof in herbicide resistance of plants, wherein the amino acid sequence of the ACCase mutant gene coding protein has the following mutations: which is mutated at position 1731 or/and 2276 corresponding to the amino acid sequence of wild-type rice ACCase. The ACCase mutant gene is introduced into the rice genome sensitive to herbicide by a transgenic method and is expressed, and the receptor plant can have resistance (tolerance) to the herbicide such as acetyl coenzyme A carboxylase inhibitor. The ACCase mutant gene can be widely applied to herbicide-resistant breeding of plants.

Description

Rice ACCase mutant type gene and application thereof in herbicide resistance of plants

Technical Field

The invention relates to a rice ACCase mutant gene and application thereof in plant herbicide resistance, belonging to the fields of plant genetic engineering and plant herbicide resistance breeding, wherein the amino acid sequence of a protein coded by the gene is mutated at position 1731 or/and 2276 of the amino acid sequence of wild rice ACCase.

Background

Acetyl coenzyme A carboxylase (ACCase) inhibitors are super-efficient herbicides, have excellent control effects on gramineous weeds, are highly safe for dicotyledonous plants, and are most commonly used in broad-leaved crop fields at present. But simultaneously, the single use of the herbicide for a long time is found, the dosage of the herbicide is generally increased year by year, the pesticide effect is reduced, namely, a large amount of sensitive weeds are reduced, but non-sensitive drug-resistant weeds gradually rise and evolve into community dominant species and are easy to induce the generation of drug-resistant weeds.

ACCase is a key enzyme in plant fatty acid synthesis, catalyzing ATP-dependent acetyl-coa carboxylation to induce a key metabolite of fatty acid and flavonoid biosynthesis (malonyl-coa), which is the first key step in fatty acid biosynthesis in plants, and then forming fatty acids under the action of fatty acid synthase, and then unsaturated fatty acids. If the synthesis of malonyl-CoA is stopped, inhibition of ACCase will cause the plant to lose this intermediate, resulting in disruption of membrane structure, enhanced permeability, and cell destruction. In plants, ACCase is localized in the cytoplasm and chloroplasts, and there are two forms of isozymes: cytoplasmic multifunctional ACCase (MF-ACCase) and chloroplast multi-subunit carboxylase ACCase (MS-ACCase). The cytoplasmic multifunctional ACCase is similar to ACCase in animal and yeast, also called eukaryotic type ACCase or ACCase I, and the enzyme in this form is a homodimer containing 2 subunits and has a molecular weight of about 500 kDa. The chloroplast multi-subunit carboxylase ACCase is similar to ACCase in E.coli and is also known as prokaryotic ACCase or ACCase II, in the form of a tetramer of 4 subunits with a molecular weight of about 700 kDa. Prokaryotic ACCase is composed of biotin carboxylation carrier protein, biotin carboxylase, acetyl coenzyme A Carboxylation Transferase (CT), wherein the beta subunit of CT is encoded by chloroplast genes, and the rest are all encoded by nuclear genes; eukaryotic ACCase consists of only a single multifunctional protein and is encoded by a nuclear gene. ACCase activity is regulated by ATP and is dependent on light. Under the condition of illumination, the ATP content is high, and the ACCase activity is increased through photosynthetic phosphorylation; during dark treatment, ATP content rapidly decreases, and ACCase activity also decreases.

ACCase is a target enzyme of highly effective herbicides like acetolactate synthase (ALS) and polyphenol oxidase (PPO), and herbicides against this target are mainly aryloxyphenoxypropionic Acid (AOPP) and Cyclohexenone (CHD) herbicides, which are highly selective for dicotyledons, and have low mammalian toxicity and good environmental characteristics, and are receiving increasing attention. The AOPP herbicide is a famous high-activity herbicide for preventing and killing gramineous weeds, and is a herbicide type which is developed rapidly and continuously to develop new varieties. This class of herbicides was first studied in the 60's of the 20 th century. The current commercialized AOPP herbicides include diclofop-methyl, stable herbicide, high-efficiency grass-cover, grass-P-ethyl, oxadixyl, and the like. The herbicide has stereoisomerism, and only the D-isomer of the isomer has obvious activity on ACCase. After the Gelidium japonicum is treated, the activity of the lipoxygenase of the plants is increased, the oxidation of unsaturated fatty acid is intensified, and the damage of free radicals to the membrane is deepened; applying various exogenous lipoxygenase inhibitors and scavenging free radical substances can reverse the toxic action of the diclofop-methyl and the galingale, and inhibit the action of the diclofop-methyl inducing the formation of ethylene, which indicates that oxygen stress is an important mechanism of the action of AOPP herbicides.

Due to the widespread use of chemical herbicides, weed resistance occurs at such a high rate and over such a wide range, not only increasing the cost of weeding, but also increasing environmental stress. Currently, there are two views of the cause of the development of resistant weed populations: a population is considered to have improved weed detoxification ability against herbicides or herbicide resistance or altered gene expression at the site of herbicide action when weeds are exposed to herbicides for a prolonged period of time. In general, the development of weed resistance is essentially a process by which a resistance gene is selected, and the inheritance of the resistance gene is the basis for the development of resistance by weeds. Previous reports suggest that resistance occurs not only because plants alter the uptake and transport of herbicides or plants accelerate herbicide metabolism, but also primarily because the site of action of herbicides is altered. A significant proportion of weeds acquire resistance by deregulating or reducing their affinity for herbicides through a change in the target enzyme ACCase. The resistance of ryegrass helveticus to diclofop-methyl is due to a partial ACCase gene mutation. At present, no patent that can resist Geranium herbicide after mutation of ACCase site in rice is reported.

Disclosure of Invention

The technical problem is as follows: the technical problem to be solved by the invention is to provide a mutant protein capable of endowing plants with herbicide resistance. Further, the technical problem to be solved by the present invention is to provide a rice ACCase mutant protein which can confer resistance to (tolerance to) acetyl-coa carboxylase herbicides on plants.

Therefore, the invention provides a nucleic acid or a gene for coding the ACCase mutant protein of the rice.

In one embodiment, the invention provides a rice ACCase mutant protein, wherein the ACCase mutant protein has any one or more of the following mutations in the amino acid sequence: it has mutation at position 1731, or 2276, or mutation at positions 1731 and 2276, respectively, corresponding to the amino acid sequence of ACCase of rice.

Preferably, the amino acid at the 1731 th position of the amino acid sequence of the rice ACCase of the invention is mutated from arginine to cysteine.

Preferably, the amino acid sequence 2276 th amino acid of the rice ACCase of the present invention is mutated from tyrosine to aspartic acid.

Preferably, the amino acid sequence 1731 th amino acid of the rice ACCase of the present invention is mutated from arginine to cysteine, and at the same time, the amino acid sequence 2276 nd amino acid is mutated from tyrosine to aspartic acid.

Preferably, the amino acid sequence 1731 of the rice ACCase of the present invention is mutated to another amino acid except arginine.

Preferably, the amino acid 2276 th amino acid of the rice ACCase of the present invention is mutated to another amino acid except tyrosine.

Preferably, the amino acid sequence 1731 of the rice ACCase of the present invention is mutated to an amino acid other than arginine, and the amino acid sequence 2276 of the rice ACCase of the present invention is mutated to an amino acid other than tyrosine.

Preferably, the mutated amino acids of the ACCase mutant protein of the present invention include, but are not limited to, the above mutations, and the mutation of the above sites into other amino acids is also within the scope of the present invention, such as: the 1731 th amino acid is mutated from arginine into alanine, glycine, phenylalanine, serine, tyrosine, proline, threonine, isoleucine, aspartic acid, glutamine, tryptophan, lysine, histidine, methionine, leucine, valine and glutamic acid; 2276 th amino acid is mutated from tyrosine to alanine, glycine, phenylalanine, serine, cysteine, proline, threonine, isoleucine, aspartic acid, glutamine, tryptophan, lysine, histidine, methionine, leucine, valine, glutamic acid, and arginine.

The invention also provides a rice ACCase mutant protein, which comprises the following components:

(a) the amino acid sequence is selected from the group consisting of SEQ ID NO.2, SEQ ID NO.4 and SEQ ID NO. 6; or

(b) And (b) a protein derived from (a) having acetyl-CoA carboxylase activity, wherein the amino acid sequence in (a) is substituted and/or deleted and/or one or more amino acids are added.

The invention also provides a nucleic acid or gene encoding the protein.

The nucleic acid or gene of the present invention comprises:

(i) nucleic acids or genes encoding said proteins; or

(ii) A nucleotide sequence which hybridizes under stringent conditions with the nucleotide sequence defined in (a) and encodes a protein having acetyl-coa carboxylase activity; or

(iii) The nucleotide sequence is selected from the group consisting of SEQ ID NO 1, SEQ ID NO:3 and SEQ ID NO: 5.

The stringent conditions are as follows: high stringency hybridization conditions were performed by washing membranes for 15 minutes at 65 ℃ in a 0.1 XSSC, 0.1% SDS solution.

Further, the rice ACCase mutant gene of the invention has one or more of the following mutations: the mutation is only generated at the 5191 th nucleotide of the rice ACCase gene, or only generated at the 6826 th nucleotide of the rice ACCase gene, or the mutation is simultaneously generated at the 5191 th nucleotide and the 6826 th nucleotide of the rice ACCase gene.

Preferably, the rice ACCase mutant gene of the present invention has one or more of the following mutations: the mutation is only from C to T at the 5191 th nucleotide of the rice ACCase gene, or from T to G at the 6826 th nucleotide of the rice ACCase gene, or from C to T at the 5191 th nucleotide and from T to G at the 6826 th nucleotide of the rice ACCase gene.

The nucleotide mutated in the ACCase mutant gene of the present invention includes, but is not limited to, the above mutations, and the mutation of the above site into other nucleotides is also within the protection scope of the present invention, for example: the A mutation is T, C, G; or the C mutation is T, A, G; or the T mutation is A, C, G; or the G mutation is T, C, A, etc. are all within the protection scope of the invention.

The invention also provides an expression cassette, a recombinant vector or a cell, which contains the nucleic acid or the gene.

The invention also provides application of the rice ACCase mutant protein, nucleic acid or gene, expression cassette, recombinant vector or cell in the aspect of herbicide resistance of plants.

Wherein the herbicide is one or more of diclofop-methyl and galingale.

The present invention also provides a method for obtaining a plant having herbicide resistance, comprising the steps of:

1) allowing the plant to comprise the nucleic acid or gene; or

2) Allowing the plants to express the rice ACCase mutant protein.

Further, the method of obtaining a plant having herbicide resistance includes transgenosis, crossing, or backcrossing.

The present invention also provides a method of identifying a plant, wherein the plant is a plant comprising the nucleic acid or gene, a plant expressing the protein, or a plant obtained by the method, comprising the steps of:

1) determining whether said plant comprises said nucleic acid or gene; or the like, or, alternatively,

2) determining whether said plant expresses said protein.

The invention also provides a herbicide protectant, which comprises the rice ACCase mutant protein.

The present invention also provides a method of controlling weeds comprising: applying an effective dose of a herbicide to a field in which a crop is grown, said plant comprising said nucleic acid or gene or said expression cassette, recombinant vector or cell, said herbicide being one or more of diclofop-methyl and galaxacum.

The present invention also provides a method for protecting a plant from damage caused by a herbicide, comprising: applying an effective dose of herbicide to a field where crops are planted, wherein the plants contain the nucleic acid or the gene or the expression cassette and the recombinant vector are introduced into the plants, and the introduced plants generate herbicide resistance protein, and the herbicide is one or more of diclofop and galium.

Further, the wild type gene of the present invention is derived from rice Jiahua No. 1.

Further, the wild type plant of Jiahua No.1 of the present invention has no resistance to Gelidone herbicides, and the transgenic plant of the present invention containing the ACCase mutation site can impart increased tolerance to acetyl-CoA carboxylase herbicides to rice plants.

The numbers of the mutant gene sequences of Jiahua No.1 and the corresponding nucleotide and amino acid sequences are as follows: the 5191 th nucleotide of the ACCase gene of Jiahua No.1 is mutated from C to T, the 1731 th amino acid is changed from arginine to cysteine, the nucleotide sequence is shown as SEQ ID No.1, and the amino acid sequence is shown as SEQ ID No. 2;

wherein, the 6826 th nucleotide of the ACCase gene of Jiahua No.1 is changed from T to G, the 2276 th amino acid is changed from tyrosine to aspartic acid, the nucleotide sequence is shown as SEQ ID No.3, and the amino acid sequence is shown as SEQ ID No. 4;

wherein, the 5191 st nucleotide of the ACCase gene of Jiahua No.1 is mutated from C to T and the 6826 th nucleotide is mutated from T to G, the 1731 st amino acid is changed from arginine to cysteine and the 2276 st amino acid is changed from tyrosine to aspartic acid, the nucleotide sequence is shown as SEQ ID No.5, and the amino acid sequence is shown as SEQ ID No. 6;

wherein, the nucleotide sequence of the wild type ACCase gene of Jiahua No.1 is shown as SEQ ID No. 7, and the amino acid sequence thereof is shown as SEQ ID No. 8.

The invention also provides methods of obtaining plants that are herbicide resistant. Further, the present invention provides methods for enhancing the tolerance of plants, plant tissues, plant cells to at least one herbicide that interferes with the activity of the ACCase enzyme. Thus, herbicide resistance in the present invention refers to resistance to (tolerance to) acetyl-coa carboxylase herbicides.

The method of enhancing the resistance of a plant, plant tissue or plant cell to a herbicide of the present invention may be carried out by transformation or by crossing, selfing and asexual propagation, or by site-directed mutagenesis of a gene, such that the altered plant comprises one or more of the nucleotide sequences of SEQ ID No.1, SEQ ID No.3 and SEQ ID No.5 of the present invention.

The method of enhancing the resistance of a plant, plant tissue or plant cell to a herbicide of the present invention can be carried out by transformation or by crossing, selfing and asexual propagation, or by site-directed mutagenesis of a gene, so that the altered plant expresses one or more of the amino acid sequences SEQ ID No.2, SEQ ID No.4 and SEQ ID No.6 of the present invention.

The invention also provides a herbicide-tolerant rice plant, which expresses the ACCase gene containing herbicide-resistant mutation sites, wherein the gene sequence is different from the ACCase gene sequence of a wild-type rice plant and is a sequence of the ACCase gene containing herbicide-resistant mutation sites. The rice ACCase mutant sequence carrying the mutant site is obtained by the inventor through a directional mutation method and is transferred into a wild rice variety, and the fact that the transgenic rice carrying the ACCase mutant site can resist the ACCase herbicide is found, while the wild rice is sensitive to the ACCase herbicide. The invention also provides the application of the nucleic acid or the gene and the protein in plant breeding for cultivating plants with herbicide resistance, especially crops, and also provides the application of the protein and the coding gene thereof in transgenic or non-transgenic plants such as rice and the like.

Has the advantages that: compared with the prior art, the invention has the following advantages:

1) according to the invention, experimental research is carried out for the first time to obtain that the wild rice is the mutation at the 1731 st position of the amino acid sequence of the rice ACCase of the indica type conventional rice seed Jiahua No.1, or the mutation at the 2276 st position corresponding to the amino acid sequence of the wild rice ACCase is carried out, or the mutation at the 1731 st position and the 2276 st position corresponding to the amino acid sequence of the wild rice ACCase are simultaneously carried out; the mutant protein obtained by the gene mutation has obvious resistance function to herbicides, in particular to acetyl coenzyme A carboxylase herbicides.

2) The ACCase mutant sequence is transferred into wild rice, and the transgenic rice obtains the resistance of ACCase enzyme herbicides. The experimental result of spraying ACCase inhibitor herbicide weed-killing in the field shows that the plant still grows normally and is fruited after the transgenic rice containing the ACCase mutant protein of the rice is applied with the acetyl coenzyme A carboxylase inhibitor herbicide with the concentration 1 time that the herbicide is recommended to be used, while the growth of the plant is stopped gradually after the wild-type rice seedling is applied with the acetyl coenzyme A carboxylase inhibitor herbicide with the concentration 1 time that the herbicide is recommended to be used, the leaves gradually turn green and light yellow, the plant cannot be pulled out, and finally the whole plant dies.

Drawings

FIG. 1 shows a transgenic rice plant containing ACCase mutation sites resistant to acetyl-CoA carboxylase inhibitor herbicides, wherein 1 represents a mutant.

FIG. 2 shows the result of PCR amplification of the entire ACCase gene containing the rice ACCase mutation site; lane 1 is Marker; the Marker has a molecular weight of 15kb, 8kb, 5kb, 3kb, 2kb, 1.5kb, 1kb, 750bp, and 500bp from top to bottom, wherein Lane 2 is DNA of wild type Jiahua No.1, Lane 3 is DNA containing herbicide-resistant mutation site ACCase-5191, Lane 4 is DNA containing herbicide-resistant mutation site ACCase-6826, and Lane 5 is DNA containing both herbicide-resistant mutation site ACCase-5191 and ACCase-6826.

FIG. 3 shows a rice PCR assay of mutant ACCase genes; there are 8 lanes in total, lane 1 is Marker; the Marker molecular weight is 15kb, 8kb, 5kb, 3kb, 2kb, 1.5kb, 1kb, 750bp and 500bp from top to bottom in sequence; lane 2 is non-transgenic wild-type DNA, lane 3 is plasmid DNA, lane 4 is transgenic plant DNA transformed with an ACCase-5191 herbicide-resistant mutant ACCase gene, lane 5 is transgenic plant DNA transformed with an ACCase-6826 herbicide-resistant mutant ACCase gene, and lane 6 is transgenic plant DNA transformed with ACCase-5191 and ACCase-6826 herbicide-resistant mutant ACCase genes, as a negative control, and lane 3 is a positive control.

Detailed Description

Embodiments of the present invention will be described in detail below with reference to examples, but those skilled in the art will appreciate that the following examples are only illustrative of the present invention and should not be construed as limiting the scope of the present invention. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products commercially available.

Example 1 Rice M3 mutant AcetylCoA carboxylase-like herbicide (Geranion)

150kg of japonica rice conventional rice seed Jiahua No.1 (purchased from agricultural germplasm resource protection and utilization platform in Jiangsu province) (M0, soaked in clear water for 2 hours) is soaked in 0.5-1.0% (w/w) Ethyl Methanesulfonate (EMS) for 6 times at room temperature for 6-9 hours, and the seeds are shaken every 1 hour in the process; the EMS solution was discarded and the seeds were soaked 5 times for 5 minutes each time with tap water tumbling, then rinsed overnight with tap water, sown the next day in the field and subjected to conventional fertilizer and water management (here M1). After the plants are mature, the seeds are mixed and harvested, aired and stored in winter. Sowing the seeds in the field the next year. When the rice seedlings (M2) grow to 3-4 leaf stage, 1.5mL of Galenon/L water is sprayed, and after 30 days, normal green plants are obtained, namely the rice mutant resisting acetyl coenzyme A carboxylase herbicides (figure 1). Seeds harvested from the Jiahua No.1 herbicide-resistant mutant are sown and emerged (M3), and 1.5mL of Geranium/L water (1 time concentration) is sprayed when M3 rice seedlings grow to 3-4 leaf stage. After the herbicide is sprayed for 15 days, the resistant seedlings M3 are normally green and can continuously grow to 20-30cm, while the non-resistant seedlings lose green leaves and even partially withered and yellow leaves, and the plants do not grow to 5-9 cm. After 30 days, the M3 resistant plants all appeared as normal green plants, while the wild type rice sprayed with the same concentration of herbicide all died, indicating that the mutant rice was resistant to at least 1-fold the concentration of the Geranium herbicide (Table 1).

Example 2: JH-7 mutation site analysis of rice acetyl coenzyme A carboxylase herbicide resistant mutant JH-7

Selecting leaves of the obtained herbicide-resistant rice mutant plant JH-7, extracting genome DNA, and sending the genome DNA to Nanjing, Yizhiyi Biotech Co. The sequencing result is compared with the wild type ACCase gene of Jiahua No.1, and the mutation of 1 site is found on the ACCase gene, the base of the 5191 site is mutated, and the C is changed into the T, so that the 1731 site of the corresponding coded amino acid sequence is changed from arginine to cysteine, namely, the nucleotide sequence of the ACCase gene of the herbicide-resistant mutant is shown as SEQ ID NO.1, and the amino acid sequence of the coded ACCase protein is shown as SEQ ID NO. 2. The rice mutant is classified and named as rice seed JH-7(Oryza sativa Japonica Group JH-7), and the strain is preserved in China Center for Type Culture Collection (CCTCC) in 2018, 9 and 28 months, and the address: wuhan university collection center (opposite to the first subsidiary school of Wuhan university) in Wuchang district, Wuhan city, Hubei province, zip code: 430072, preservation number is CCTCC No: and P201815.

Example 3: JH-8 mutant site analysis of rice acetyl coenzyme A carboxylase herbicide resistant mutant

Selecting a plurality of mutant leaves from herbicide-resistant rice mutant plants JH-8, respectively extracting genome DNA, and sending the genome DNA to Nanjing, a Biotech limited company for genome sequencing. The sequencing result is compared with the wild type ACCase gene of Jiahua No.1, and the mutation of 1 site is found on the ACCase gene, the base of the 6826 site is mutated, and is changed from T to G, so that the 2276 site of the corresponding coded amino acid sequence is changed from tyrosine to aspartic acid, namely, the nucleotide sequence of the ACCase gene of the herbicide-resistant mutant is shown as SEQ ID NO.3, and the amino acid sequence of the coded ACCase protein is shown as SEQ ID NO. 4. The rice mutant is classified and named as rice seed JH-8(Oryza sativa Japonica Group JH-8), and the strain is preserved in China Center for Type Culture Collection (CCTCC) in 2018, 9 and 28 months, and the address: wuhan university collection center (opposite to the first subsidiary school of Wuhan university) in Wuchang district, Wuhan city, Hubei province, zip code: 430072, preservation number is CCTCC No: and P201816.

Example 4: JH-9 mutant site analysis of rice acetyl coenzyme A carboxylase herbicide resistant mutant

Selecting a plurality of mutant leaves from herbicide-resistant rice mutant plants JH-9, respectively extracting genome DNA, and sending the genome DNA to Nanjing, a Biotech limited company for genome sequencing. The sequencing result is compared with the wild type ACCase gene of Jiahua No.1, and the mutation of 2 sites, the base mutation of 5191 site, C to T, and the base mutation of 6826 site to G, which results in the conversion of arginine to cysteine at 1731 site and tyrosine to aspartic acid at 2276 site of the corresponding coded amino acid sequence, are found on the ACCase gene, i.e. the nucleotide sequence of the ACCase gene of the herbicide-resistant mutant is shown in SEQ ID No.5, and the amino acid sequence of the coded ACCase protein is shown in SEQ ID No. 6. The rice mutant is classified and named as rice seed JH-9(Oryza sativa Japonica Group JH-9), and the strain is preserved in China Center for Type Culture Collection (CCTCC) in 2018, 9 and 28 months, and the address: wuhan university collection center (opposite to the first subsidiary school of Wuhan university) in Wuchang district, Wuhan city, Hubei province, zip code: 430072, preservation number is CCTCC No: p201817.

Example 5: transgenic ACCase rice anti-Galenon herbicide

Design of specific primer 5-CCCGGGATGACATCCACACATGTGGCGACAT-3' and 5-GGGTACCCTCAACCAAGGACCTTCCTGATTTCT-3 ', 5' of which has SmaI and KpnI restriction modification sites added. Referring to the method of example 3, the mutant ACCase gene was amplified from the genomic DNA of the above-mentioned rice mutant, Kaffir No.1 herbicide-resistant mutant, and after sequencing was completed correctly, the mutant ACCase gene fragment and the plant expression vector pCAMBIA1301 plasmid (available from pCAMBIA) were double-digested with SmaI and KpnI, respectively, and the digested products were ligated with T4-DNase (available from TaKaRa), and the ligated products were transformed into E.coli. The recombinant plasmid extracts DNA, and double restriction enzyme digestion verification with SmaI and KpnI can generate a large plasmid fragment and a small gene fragment (figure 2), and proves that the ACCase gene with the nucleotide sequence shown as SEQ ID NO.1, SEQ ID NO.3 and SEQ ID NO.5 is cloned into a plant expression vector pCAMBIA1301 plasmid (purchased from pCAMBIA). Transforming the constructed plasmid vector into a farm poleThe strain EHA105 was cultured. Japonica rice Nipponbare (purchased from Jiangsu province agriculture germplasm resource protection and utilization platform) is transformed by adopting a conventional agrobacterium-mediated method, and after a transgenic plant is harvested, the transgenic plant is detected by PCR when the progeny plant grows to 3-4 leaf stage (figure 3). The PCR detection primer is a forward primer 35SF5 ' -ATGGTTAGAGAGGCTTACGC-3 ', a reverse primer 5R5 ' -AGCAACAGGTCAGCCTTATCCAC-3 ', and the amplified fragment comprises a CaMV35S promoter and a 5 ' end sequence of an ACCase gene and has the size of about 2 kb. PCR amplification reaction System referring to example 3, the PCR amplification reaction procedure used a two-step procedure, annealing and extension were combined together using 68 ℃. The amplification procedure was as follows: pre-denaturation: 3min at 98 ℃; 30 cycles: denaturation at 98 ℃ for 10 sec; extending at 68 ℃ for 2 min; and (3) heat preservation: 10min at 72 ℃. After positive PCR identification, 1.5mL of Galenon/L water (1 time of recommended use concentration) was sprayed to transgenic rice and wild type rice seedlings, and after 30 days, the transgenic rice was found to have a good growth state, while the non-transgenic Nipponbare rice was completely died (Table 2).

TABLE 1 injury ratio of acetyl-CoA carboxylase herbicides to wild type and M3 mutant

TABLE 2 injury rates of acetyl-CoA carboxylase herbicides on wild-type and transgenic rice

While specific embodiments of the invention have been described in detail, it will be appreciated by those skilled in the art that various modifications and alternatives to those details could be developed in light of the overall teachings of the disclosure, and are within the scope of the invention. The full scope of the invention is given by the appended patent claims and any equivalents thereof.

Sequence listing

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<120> ACCase mutant protein imparting herbicide resistance to plants and use thereof

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gctgtttggc ctggttgggg tcatgcatct gagaatcctg aacttccaga tgcgctgact 720

gcaaaaggaa ttgtttttct tgggccacca gcatcatcaa tgcatgcatt aggagacaag 780

gttggctcag ctctcattgc tcaagcagct ggagttccaa cacttgcttg gagtggatca 840

catgtggaag ttcctctgga gtgttgcttg gactcaatac ctgatgagat gtatagaaaa 900

gcttgtgtta ctaccacaga ggaagcagtt gcaagttgtc aggtggttgg ttatcctgcc 960

atgattaagg catcttgggg tggtggtggt aaaggaataa ggaaggttca taatgatgat 1020

gaggttagga cattatttaa gcaagttcaa ggcgaagtac ctggttcccc aatatttatc 1080

atgaggctag ctgctcagag tcgacatctt gaagttcagt tgctttgtga tcaatatggc 1140

aacgtagcag cacttcacag tcgagattgc agtgtacaac ggcgacacca aaagataatc 1200

gaggaaggac cagttactgt tgctcctcgt gagactgtga aagagcttga gcaggcagca 1260

cggaggcttg ctaaagctgt gggttatgtt ggtgctgcta ctgttgaata cctttacagc 1320

atggaaactg gtgaatatta ttttctggaa cttaatccac ggctacaggt tgagcatcct 1380

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ataccccttt ggcagattcc agagatcagg cgcttctacg gaatgaacca tggaggaggc 1500

tatgaccttt ggaggaaaac agcagctcta gcgactccat ttaactttga tgaagtagat 1560

tctaaatggc caaaaggcca ctgcgtagct gttagaataa ctagcgagga tccagatgat 1620

gggtttaagc ctactggtgg aaaagtaaag gagataagtt tcaagagtaa accaaatgtt 1680

tgggcctatt tctcagtaaa gtctggtgga ggcatccatg aattcgctga ttctcagttc 1740

ggacatgttt ttgcgtatgg aactactaga tcggcagcaa taactaccat ggctcttgca 1800

ctaaaagagg ttcaaattcg tggagaaatt cattcaaacg tagactacac agttgaccta 1860

ttaaatgcct cagattttag agaaaataag attcatactg gttggctgga taccaggata 1920

gccatgcgtg ttcaagctga gaggcctcca tggtatattt cagtcgttgg aggggcttta 1980

tataaaacag taactgccaa cacggccact gtttctgatt atgttggtta tcttaccaag 2040

ggccagattc caccaaagca tatatccctt gtctatacga ctgttgcttt gaatatagat 2100

gggaaaaaat atacaatcga tactgtgagg agtggacatg gtagctacag attgcgaatg 2160

aatggatcaa cggttgacgc aaatgtacaa atattatgtg atggtgggct tttaatgcag 2220

ctggatggaa acagccatgt aatttatgct gaagaagagg ccagtggtac acgacttctt 2280

attgatggaa agacatgcat gttacagaat gaccatgacc catcaaagtt attagctgag 2340

acaccatgca aacttcttcg tttcttggtt gctgatggtg ctcatgttga tgctgatgta 2400

ccatatgcgg aagttgaggt tatgaagatg tgcatgcccc tcttatcacc cgcttctggt 2460

gtcatacatg ttgtaatgtc tgagggccaa gcaatgcagg ctggtgatct tatagctagg 2520

ctggatcttg atgacccttc tgctgttaag agagctgagc cgttcgaaga tacttttcca 2580

caaatgggtc tccctattgc tgcttctggc caagttcaca aattatgtgc tgcaagtctg 2640

aatgcttgtc gaatgatcct tgcggggtat gagcatgata ttgacaaggt tgtgccagag 2700

ttggtatact gcctagacac tccggagctt cctttcctgc agtgggagga gcttatgtct 2760

gttttagcaa ctagacttcc aagaaatctt aaaagtgagt tggagggcaa atatgaggaa 2820

tacaaagtaa aatttgactc tgggataatc aatgatttcc ctgccaatat gctacgagtg 2880

ataattgagg aaaatcttgc atgtggttct gagaaggaga aggctacaaa tgagaggctt 2940

gttgagcctc ttatgagcct actgaagtca tatgagggtg ggagagaaag tcatgctcac 3000

tttgttgtca agtccctttt tgaggagtat ctctatgttg aagaattgtt cagtgatgga 3060

attcagtctg atgtgattga gcgtctgcgc cttcaacata gtaaagacct acagaaggtc 3120

gtagacattg tgttgtccca ccagagtgtt agaaataaaa ctaagctgat actaaaactc 3180

atggagagtc tggtctatcc aaatcctgct gcctacaggg atcaattgat tcgcttttct 3240

tcccttaatc acaaagcgta ttacaagttg gcacttaaag ctagtgaact tcttgaacaa 3300

acaaaactta gtgagctccg tgcaagaata gcaaggagcc tttcagagct ggagatgttt 3360

actgaggaaa gcaagggtct ctccatgcat aagcgagaaa ttgccattaa ggagagcatg 3420

gaagatttag tcactgctcc actgccagtt gaagatgcgc tcatttcttt atttgattgt 3480

agtgatacaa ctgttcaaca gagagtgatt gagacttata tagctcgatt ataccagcct 3540

catcttgtaa aggacagtat caaaatgaaa tggatagaat cgggtgttat tgctttatgg 3600

gaatttcctg aagggcattt tgatgcaaga aatggaggag cggttcttgg tgacaaaaga 3660

tggggtgcca tggtcattgt caagtctctt gaatcacttt caatggccat tagatttgca 3720

ctaaaggaga catcacacta cactagctct gagggcaata tgatgcatat tgctttgttg 3780

ggtgctgata ataagatgca tataattcaa gaaagtggtg atgatgctga cagaatagcc 3840

aaacttccct tgatactaaa ggataatgta accgatctgc atgcctctgg tgtgaaaaca 3900

ataagtttca ttgttcaaag agatgaagca cggatgacaa tgcgtcgtac cttcctttgg 3960

tctgatgaaa agctttctta tgaggaagag ccaattctcc ggcatgtgga acctcctctt 4020

tctgcacttc ttgagttgga caagttgaaa gtgaaaggat acaatgaaat gaagtatacc 4080

ccatcacggg atcgtcaatg gcatatctac acacttagaa atactgaaaa ccccaaaatg 4140

ttgcaccggg tatttttccg aacccttgtc aggcaaccca gtgtatccaa caagttttct 4200

tcgggccaga ttggtgacat ggaagttggg agtgctgaag aacctctgtc atttacatca 4260

accagcatat taagatcttt gatgactgct atagaggaat tggagcttca cgcaattaga 4320

actggccatt cacacatgta tttgcatgta ttgaaagaac aaaagcttct tgatcttgtt 4380

ccagtttcag ggaatacagt tttggatgtt ggtcaagatg aagctactgc atattcactt 4440

ttaaaagaaa tggctatgaa gatacatgaa cttgttggtg caagaatgca ccatctttct 4500

gtatgccaat gggaagtgaa acttaagttg gactgcgatg gtcctgccag tggtacctgg 4560

aggattgtaa caaccaatgt tactagtcac acttgcactg tggatatcta ccgtgagatg 4620

gaagataaag aatcacggaa gttagtatac catcccgcca ctccggcggc tggtcctctg 4680

catggtgtgg cactgaataa tccatatcag cctttgagtg tcattgatct caaacgctgt 4740

tctgctagga ataatagaac tacatactgc tatgattttc cactggcatt tgaaactgca 4800

gtgaggaagt catggtcctc tagtacctct ggtgcttcta aaggtgttga aaatgcccaa 4860

tgttatgtta aagctacaga gttggtattt gcggacaaac atgggtcatg gggcactcct 4920

ttagttcaaa tggaccggcc tgctgggctc aatgacattg gtatggtagc ttggaccttg 4980

aagatgtcca ctcctgaatt tcctagtggt agggagatta ttgttgttgc aaatgatatt 5040

acgttcagag ctggatcatt tggcccaagg gaagatgcat tttttgaagc tgttaccaac 5100

ctagcctgtg agaagaaact tcctcttatt tatttggcag caaattctgg tgctcgaatt 5160

ggcatagcag atgaagtgaa atcttgcttc tgtgttgggt ggtctgatga tggcagccct 5220

gaacgtgggt ttcagtacat ttatctaagc gaagaagact atgctcgtat tggcacttct 5280

gtcatagcac ataagatgca gctagacagt ggtgaaatta ggtgggttat tgattctgtt 5340

gtgggcaagg aagatggact tggtgtggag aatatacatg gaagtgctgc tattgccagt 5400

gcttattcta gggcatataa ggagacattt acacttacat ttgtgactgg aagaactgtt 5460

ggaataggag cttatcttgc tcgacttggc atccggtgca tacagcgtct tgaccagcct 5520

attattctta caggctattc tgcactgaac aagcttcttg ggcgggaagt gtacagctcc 5580

cacatgcagt tgggtggtcc caaaatcatg gcaactaatg gtgttgtcca tcttactgtt 5640

tcagatgacc ttgaaggcgt ttctaatata ttgaggtggc tcagttatgt tcctgcctac 5700

attggtggac cacttccagt aacaacaccg ttggacccac cggacagacc tgttgcatac 5760

attcctgaga actcgtgtga tcctcgagcg gctatccgtg gtgttgatga cagccaaggg 5820

aaatggttag gtggtatgtt tgataaagac agctttgtgg aaacatttga aggttgggct 5880

aagacagtgg ttactggcag agcaaagctt ggtggaattc cagtgggtgt gatagctgtg 5940

gagactcaga ccatgatgca aactatccct gctgaccctg gtcagcttga ttcccgtgag 6000

caatctgttc ctcgtgctgg acaagtgtgg tttccagatt ctgcaaccaa gactgcgcag 6060

gcattgctgg acttcaaccg tgaaggatta cctctgttca tcctcgctaa ctggagaggc 6120

ttctctggtg gacaaagaga tctttttgaa ggaattcttc aggctggctc gactattgtt 6180

gagaacctta ggacatacaa tcagcctgcc tttgtctaca ttcccatggc tgcagagcta 6240

cgaggagggg cttgggttgt ggttgatagc aagataaacc cagaccgcat tgagtgctat 6300

gctgagagga ctgcaaaagg caatgttctg gaaccgcaag ggttaattga gatcaagttc 6360

aggtcagagg aactccagga ttgcatgagt cggcttgacc caacattaat tgatctgaaa 6420

gcaaaactcg aagtagcaaa taaaaatgga agtgctgaca caaaatcgct tcaagaaaat 6480

atagaagctc gaacaaaaca gttgatgcct ctatatactc agattgcgat acggtttgct 6540

gaattgcatg atacatccct cagaatggct gcgaaaggtg tgattaagaa agttgtggac 6600

tgggaagaat cacgatcttt cttctataag agattacgga ggaggatctc tgaggatgtt 6660

cttgcaaaag aaattagagc tgtagcaggt gagcagtttt cccaccaacc agcaatcgag 6720

ctgatcaaga aatggtattc agcttcacat gcagctgaat gggatgatga cgatgctttt 6780

gttgcttgga tggataaccc tgaaaactac aaggattata ttcaatatct taaggctcaa 6840

agagtatccc aatccctctc aagtctttca gattccagct cagatttgca agccctgcca 6900

cagggtcttt ccatgttact agataagatg gatccctcta gaagagctca acttgttgaa 6960

gaaatcagga aggtccttgg ttga 6984

<210> 2

<211> 2327

<212> PRT

<213> Rice (Oryza sativa) Jiahua No.1 ACCase-5191 mutant amino acid sequence

<400> 2

MTSTHVATLG VGAQAPPRHQ KKSAGTAFVS SGSSRPSYRK NGQRTRSLRE ESNGGVSDSK 60

KLNHSIRQGL AGIIDLPNDA ASEVDISHGS EDPRGPTVPG SYQMNGIINE THNGRHASVS 120

KVVEFCTALG GKTPIHSVLV ANNGMAAAKF MRSVRTWAND TFGSEKAIQL IAMATPEDLR 180

INAEHIRIAD QFVEVPGGTN NNNYANVQLI VEIAERTGVS AVWPGWGHAS ENPELPDALT 240

AKGIVFLGPP ASSMHALGDK VGSALIAQAA GVPTLAWSGS HVEVPLECCL DSIPDEMYRK 300

ACVTTTEEAV ASCQVVGYPA MIKASWGGGG KGIRKVHNDD EVRTLFKQVQ GEVPGSPIFI 360

MRLAAQSRHL EVQLLCDQYG NVAALHSRDC SVQRRHQKII EEGPVTVAPR ETVKELEQAA 420

RRLAKAVGYV GAATVEYLYS METGEYYFLE LNPRLQVEHP VTEWIAEVNL PAAQVAVGMG 480

IPLWQIPEIR RFYGMNHGGG YDLWRKTAAL ATPFNFDEVD SKWPKGHCVA VRITSEDPDD 540

GFKPTGGKVK EISFKSKPNV WAYFSVKSGG GIHEFADSQF GHVFAYGTTR SAAITTMALA 600

LKEVQIRGEI HSNVDYTVDL LNASDFRENK IHTGWLDTRI AMRVQAERPP WYISVVGGAL 660

YKTVTANTAT VSDYVGYLTK GQIPPKHISL VYTTVALNID GKKYTIDTVR SGHGSYRLRM 720

NGSTVDANVQ ILCDGGLLMQ LDGNSHVIYA EEEASGTRLL IDGKTCMLQN DHDPSKLLAE 780

TPCKLLRFLV ADGAHVDADV PYAEVEVMKM CMPLLSPASG VIHVVMSEGQ AMQAGDLIAR 840

LDLDDPSAVK RAEPFEDTFP QMGLPIAASG QVHKLCAASL NACRMILAGY EHDIDKVVPE 900

LVYCLDTPEL PFLQWEELMS VLATRLPRNL KSELEGKYEE YKVKFDSGII NDFPANMLRV 960

IIEENLACGS EKEKATNERL VEPLMSLLKS YEGGRESHAH FVVKSLFEEY LYVEELFSDG 1020

IQSDVIERLR LQHSKDLQKV VDIVLSHQSV RNKTKLILKL MESLVYPNPA AYRDQLIRFS 1080

SLNHKAYYKL ALKASELLEQ TKLSELRARI ARSLSELEMF TEESKGLSMH KREIAIKESM 1140

EDLVTAPLPV EDALISLFDC SDTTVQQRVI ETYIARLYQP HLVKDSIKMK WIESGVIALW 1200

EFPEGHFDAR NGGAVLGDKR WGAMVIVKSL ESLSMAIRFA LKETSHYTSS EGNMMHIALL 1260

GADNKMHIIQ ESGDDADRIA KLPLILKDNV TDLHASGVKT ISFIVQRDEA RMTMRRTFLW 1320

SDEKLSYEEE PILRHVEPPL SALLELDKLK VKGYNEMKYT PSRDRQWHIY TLRNTENPKM 1380

LHRVFFRTLV RQPSVSNKFS SGQIGDMEVG SAEEPLSFTS TSILRSLMTA IEELELHAIR 1440

TGHSHMYLHV LKEQKLLDLV PVSGNTVLDV GQDEATAYSL LKEMAMKIHE LVGARMHHLS 1500

VCQWEVKLKL DCDGPASGTW RIVTTNVTSH TCTVDIYREM EDKESRKLVY HPATPAAGPL 1560

HGVALNNPYQ PLSVIDLKRC SARNNRTTYC YDFPLAFETA VRKSWSSSTS GASKGVENAQ 1620

CYVKATELVF ADKHGSWGTP LVQMDRPAGL NDIGMVAWTL KMSTPEFPSG REIIVVANDI 1680

TFRAGSFGPR EDAFFEAVTN LACEKKLPLI YLAANSGARI GIADEVKSCF CVGWSDDGSP 1740

ERGFQYIYLS EEDYARIGTS VIAHKMQLDS GEIRWVIDSV VGKEDGLGVE NIHGSAAIAS 1800

AYSRAYKETF TLTFVTGRTV GIGAYLARLG IRCIQRLDQP IILTGYSALN KLLGREVYSS 1860

HMQLGGPKIM ATNGVVHLTV SDDLEGVSNI LRWLSYVPAY IGGPLPVTTP LDPPDRPVAY 1920

IPENSCDPRA AIRGVDDSQG KWLGGMFDKD SFVETFEGWA KTVVTGRAKL GGIPVGVIAV 1980

ETQTMMQTIP ADPGQLDSRE QSVPRAGQVW FPDSATKTAQ ALLDFNREGL PLFILANWRG 2040

FSGGQRDLFE GILQAGSTIV ENLRTYNQPA FVYIPMAAEL RGGAWVVVDS KINPDRIECY 2100

AERTAKGNVL EPQGLIEIKF RSEELQDCMS RLDPTLIDLK AKLEVANKNG SADTKSLQEN 2160

IEARTKQLMP LYTQIAIRFA ELHDTSLRMA AKGVIKKVVD WEESRSFFYK RLRRRISEDV 2220

LAKEIRAVAG EQFSHQPAIE LIKKWYSASH AAEWDDDDAF VAWMDNPENY KDYIQYLKAQ 2280

RVSQSLSSLS DSSSDLQALP QGLSMLLDKM DPSRRAQLVE EIRKVLG 2327

<210> 3

<211> 6984

<212> DNA

<213> Rice (Oryza sativa) Jiahua No.1 ACCase-6826 mutant nucleotide sequence

<400> 3

atgacatcca cacatgtggc gacattggga gttggtgccc aggcacctcc tcgtcaccag 60

aaaaagtcag ctggcactgc atttgtatca tctgggtcat caagaccctc ataccgaaag 120

aatggtcagc gtactcggtc acttagggaa gaaagcaatg gaggagtgtc tgattccaaa 180

aagcttaacc actctattcg ccaaggtctt gctggcatca ttgacctccc aaatgacgca 240

gcttcagaag ttgatatttc acatggttcc gaagatccca gggggcctac ggtcccaggt 300

tcctaccaaa tgaatgggat tatcaatgaa acacataatg ggaggcatgc ttcagtctcc 360

aaggttgttg agttttgtac ggcacttggt ggcaaaacac caattcacag tgtattagtg 420

gccaacaatg gaatggcagc agctaagttc atgcggagtg tccgaacatg ggctaatgat 480

acttttggat cagagaaggc aattcagctg atagctatgg caactccgga ggatctgagg 540

ataaatgcag agcacatcag aattgccgat caatttgtag aggtacctgg tggaacaaac 600

aacaacaact atgcaaatgt ccaactcata gtggagatag cagagagaac aggtgtttct 660

gctgtttggc ctggttgggg tcatgcatct gagaatcctg aacttccaga tgcgctgact 720

gcaaaaggaa ttgtttttct tgggccacca gcatcatcaa tgcatgcatt aggagacaag 780

gttggctcag ctctcattgc tcaagcagct ggagttccaa cacttgcttg gagtggatca 840

catgtggaag ttcctctgga gtgttgcttg gactcaatac ctgatgagat gtatagaaaa 900

gcttgtgtta ctaccacaga ggaagcagtt gcaagttgtc aggtggttgg ttatcctgcc 960

atgattaagg catcttgggg tggtggtggt aaaggaataa ggaaggttca taatgatgat 1020

gaggttagga cattatttaa gcaagttcaa ggcgaagtac ctggttcccc aatatttatc 1080

atgaggctag ctgctcagag tcgacatctt gaagttcagt tgctttgtga tcaatatggc 1140

aacgtagcag cacttcacag tcgagattgc agtgtacaac ggcgacacca aaagataatc 1200

gaggaaggac cagttactgt tgctcctcgt gagactgtga aagagcttga gcaggcagca 1260

cggaggcttg ctaaagctgt gggttatgtt ggtgctgcta ctgttgaata cctttacagc 1320

atggaaactg gtgaatatta ttttctggaa cttaatccac ggctacaggt tgagcatcct 1380

gtcactgagt ggatagctga agtaaatttg cctgcggctc aagttgctgt tggaatgggt 1440

ataccccttt ggcagattcc agagatcagg cgcttctacg gaatgaacca tggaggaggc 1500

tatgaccttt ggaggaaaac agcagctcta gcgactccat ttaactttga tgaagtagat 1560

tctaaatggc caaaaggcca ctgcgtagct gttagaataa ctagcgagga tccagatgat 1620

gggtttaagc ctactggtgg aaaagtaaag gagataagtt tcaagagtaa accaaatgtt 1680

tgggcctatt tctcagtaaa gtctggtgga ggcatccatg aattcgctga ttctcagttc 1740

ggacatgttt ttgcgtatgg aactactaga tcggcagcaa taactaccat ggctcttgca 1800

ctaaaagagg ttcaaattcg tggagaaatt cattcaaacg tagactacac agttgaccta 1860

ttaaatgcct cagattttag agaaaataag attcatactg gttggctgga taccaggata 1920

gccatgcgtg ttcaagctga gaggcctcca tggtatattt cagtcgttgg aggggcttta 1980

tataaaacag taactgccaa cacggccact gtttctgatt atgttggtta tcttaccaag 2040

ggccagattc caccaaagca tatatccctt gtctatacga ctgttgcttt gaatatagat 2100

gggaaaaaat atacaatcga tactgtgagg agtggacatg gtagctacag attgcgaatg 2160

aatggatcaa cggttgacgc aaatgtacaa atattatgtg atggtgggct tttaatgcag 2220

ctggatggaa acagccatgt aatttatgct gaagaagagg ccagtggtac acgacttctt 2280

attgatggaa agacatgcat gttacagaat gaccatgacc catcaaagtt attagctgag 2340

acaccatgca aacttcttcg tttcttggtt gctgatggtg ctcatgttga tgctgatgta 2400

ccatatgcgg aagttgaggt tatgaagatg tgcatgcccc tcttatcacc cgcttctggt 2460

gtcatacatg ttgtaatgtc tgagggccaa gcaatgcagg ctggtgatct tatagctagg 2520

ctggatcttg atgacccttc tgctgttaag agagctgagc cgttcgaaga tacttttcca 2580

caaatgggtc tccctattgc tgcttctggc caagttcaca aattatgtgc tgcaagtctg 2640

aatgcttgtc gaatgatcct tgcggggtat gagcatgata ttgacaaggt tgtgccagag 2700

ttggtatact gcctagacac tccggagctt cctttcctgc agtgggagga gcttatgtct 2760

gttttagcaa ctagacttcc aagaaatctt aaaagtgagt tggagggcaa atatgaggaa 2820

tacaaagtaa aatttgactc tgggataatc aatgatttcc ctgccaatat gctacgagtg 2880

ataattgagg aaaatcttgc atgtggttct gagaaggaga aggctacaaa tgagaggctt 2940

gttgagcctc ttatgagcct actgaagtca tatgagggtg ggagagaaag tcatgctcac 3000

tttgttgtca agtccctttt tgaggagtat ctctatgttg aagaattgtt cagtgatgga 3060

attcagtctg atgtgattga gcgtctgcgc cttcaacata gtaaagacct acagaaggtc 3120

gtagacattg tgttgtccca ccagagtgtt agaaataaaa ctaagctgat actaaaactc 3180

atggagagtc tggtctatcc aaatcctgct gcctacaggg atcaattgat tcgcttttct 3240

tcccttaatc acaaagcgta ttacaagttg gcacttaaag ctagtgaact tcttgaacaa 3300

acaaaactta gtgagctccg tgcaagaata gcaaggagcc tttcagagct ggagatgttt 3360

actgaggaaa gcaagggtct ctccatgcat aagcgagaaa ttgccattaa ggagagcatg 3420

gaagatttag tcactgctcc actgccagtt gaagatgcgc tcatttcttt atttgattgt 3480

agtgatacaa ctgttcaaca gagagtgatt gagacttata tagctcgatt ataccagcct 3540

catcttgtaa aggacagtat caaaatgaaa tggatagaat cgggtgttat tgctttatgg 3600

gaatttcctg aagggcattt tgatgcaaga aatggaggag cggttcttgg tgacaaaaga 3660

tggggtgcca tggtcattgt caagtctctt gaatcacttt caatggccat tagatttgca 3720

ctaaaggaga catcacacta cactagctct gagggcaata tgatgcatat tgctttgttg 3780

ggtgctgata ataagatgca tataattcaa gaaagtggtg atgatgctga cagaatagcc 3840

aaacttccct tgatactaaa ggataatgta accgatctgc atgcctctgg tgtgaaaaca 3900

ataagtttca ttgttcaaag agatgaagca cggatgacaa tgcgtcgtac cttcctttgg 3960

tctgatgaaa agctttctta tgaggaagag ccaattctcc ggcatgtgga acctcctctt 4020

tctgcacttc ttgagttgga caagttgaaa gtgaaaggat acaatgaaat gaagtatacc 4080

ccatcacggg atcgtcaatg gcatatctac acacttagaa atactgaaaa ccccaaaatg 4140

ttgcaccggg tatttttccg aacccttgtc aggcaaccca gtgtatccaa caagttttct 4200

tcgggccaga ttggtgacat ggaagttggg agtgctgaag aacctctgtc atttacatca 4260

accagcatat taagatcttt gatgactgct atagaggaat tggagcttca cgcaattaga 4320

actggccatt cacacatgta tttgcatgta ttgaaagaac aaaagcttct tgatcttgtt 4380

ccagtttcag ggaatacagt tttggatgtt ggtcaagatg aagctactgc atattcactt 4440

ttaaaagaaa tggctatgaa gatacatgaa cttgttggtg caagaatgca ccatctttct 4500

gtatgccaat gggaagtgaa acttaagttg gactgcgatg gtcctgccag tggtacctgg 4560

aggattgtaa caaccaatgt tactagtcac acttgcactg tggatatcta ccgtgagatg 4620

gaagataaag aatcacggaa gttagtatac catcccgcca ctccggcggc tggtcctctg 4680

catggtgtgg cactgaataa tccatatcag cctttgagtg tcattgatct caaacgctgt 4740

tctgctagga ataatagaac tacatactgc tatgattttc cactggcatt tgaaactgca 4800

gtgaggaagt catggtcctc tagtacctct ggtgcttcta aaggtgttga aaatgcccaa 4860

tgttatgtta aagctacaga gttggtattt gcggacaaac atgggtcatg gggcactcct 4920

ttagttcaaa tggaccggcc tgctgggctc aatgacattg gtatggtagc ttggaccttg 4980

aagatgtcca ctcctgaatt tcctagtggt agggagatta ttgttgttgc aaatgatatt 5040

acgttcagag ctggatcatt tggcccaagg gaagatgcat tttttgaagc tgttaccaac 5100

ctagcctgtg agaagaaact tcctcttatt tatttggcag caaattctgg tgctcgaatt 5160

ggcatagcag atgaagtgaa atcttgcttc cgtgttgggt ggtctgatga tggcagccct 5220

gaacgtgggt ttcagtacat ttatctaagc gaagaagact atgctcgtat tggcacttct 5280

gtcatagcac ataagatgca gctagacagt ggtgaaatta ggtgggttat tgattctgtt 5340

gtgggcaagg aagatggact tggtgtggag aatatacatg gaagtgctgc tattgccagt 5400

gcttattcta gggcatataa ggagacattt acacttacat ttgtgactgg aagaactgtt 5460

ggaataggag cttatcttgc tcgacttggc atccggtgca tacagcgtct tgaccagcct 5520

attattctta caggctattc tgcactgaac aagcttcttg ggcgggaagt gtacagctcc 5580

cacatgcagt tgggtggtcc caaaatcatg gcaactaatg gtgttgtcca tcttactgtt 5640

tcagatgacc ttgaaggcgt ttctaatata ttgaggtggc tcagttatgt tcctgcctac 5700

attggtggac cacttccagt aacaacaccg ttggacccac cggacagacc tgttgcatac 5760

attcctgaga actcgtgtga tcctcgagcg gctatccgtg gtgttgatga cagccaaggg 5820

aaatggttag gtggtatgtt tgataaagac agctttgtgg aaacatttga aggttgggct 5880

aagacagtgg ttactggcag agcaaagctt ggtggaattc cagtgggtgt gatagctgtg 5940

gagactcaga ccatgatgca aactatccct gctgaccctg gtcagcttga ttcccgtgag 6000

caatctgttc ctcgtgctgg acaagtgtgg tttccagatt ctgcaaccaa gactgcgcag 6060

gcattgctgg acttcaaccg tgaaggatta cctctgttca tcctcgctaa ctggagaggc 6120

ttctctggtg gacaaagaga tctttttgaa ggaattcttc aggctggctc gactattgtt 6180

gagaacctta ggacatacaa tcagcctgcc tttgtctaca ttcccatggc tgcagagcta 6240

cgaggagggg cttgggttgt ggttgatagc aagataaacc cagaccgcat tgagtgctat 6300

gctgagagga ctgcaaaagg caatgttctg gaaccgcaag ggttaattga gatcaagttc 6360

aggtcagagg aactccagga ttgcatgagt cggcttgacc caacattaat tgatctgaaa 6420

gcaaaactcg aagtagcaaa taaaaatgga agtgctgaca caaaatcgct tcaagaaaat 6480

atagaagctc gaacaaaaca gttgatgcct ctatatactc agattgcgat acggtttgct 6540

gaattgcatg atacatccct cagaatggct gcgaaaggtg tgattaagaa agttgtggac 6600

tgggaagaat cacgatcttt cttctataag agattacgga ggaggatctc tgaggatgtt 6660

cttgcaaaag aaattagagc tgtagcaggt gagcagtttt cccaccaacc agcaatcgag 6720

ctgatcaaga aatggtattc agcttcacat gcagctgaat gggatgatga cgatgctttt 6780

gttgcttgga tggataaccc tgaaaactac aaggattata ttcaagatct taaggctcaa 6840

agagtatccc aatccctctc aagtctttca gattccagct cagatttgca agccctgcca 6900

cagggtcttt ccatgttact agataagatg gatccctcta gaagagctca acttgttgaa 6960

gaaatcagga aggtccttgg ttga 6984

<210> 4

<211> 2327

<212> PRT

<213> Rice (Oryza sativa) Jiahua No.1 ACCase-6826 mutant amino acid sequence

<400> 4

MTSTHVATLG VGAQAPPRHQ KKSAGTAFVS SGSSRPSYRK NGQRTRSLRE ESNGGVSDSK 60

KLNHSIRQGL AGIIDLPNDA ASEVDISHGS EDPRGPTVPG SYQMNGIINE THNGRHASVS 120

KVVEFCTALG GKTPIHSVLV ANNGMAAAKF MRSVRTWAND TFGSEKAIQL IAMATPEDLR 180

INAEHIRIAD QFVEVPGGTN NNNYANVQLI VEIAERTGVS AVWPGWGHAS ENPELPDALT 240

AKGIVFLGPP ASSMHALGDK VGSALIAQAA GVPTLAWSGS HVEVPLECCL DSIPDEMYRK 300

ACVTTTEEAV ASCQVVGYPA MIKASWGGGG KGIRKVHNDD EVRTLFKQVQ GEVPGSPIFI 360

MRLAAQSRHL EVQLLCDQYG NVAALHSRDC SVQRRHQKII EEGPVTVAPR ETVKELEQAA 420

RRLAKAVGYV GAATVEYLYS METGEYYFLE LNPRLQVEHP VTEWIAEVNL PAAQVAVGMG 480

IPLWQIPEIR RFYGMNHGGG YDLWRKTAAL ATPFNFDEVD SKWPKGHCVA VRITSEDPDD 540

GFKPTGGKVK EISFKSKPNV WAYFSVKSGG GIHEFADSQF GHVFAYGTTR SAAITTMALA 600

LKEVQIRGEI HSNVDYTVDL LNASDFRENK IHTGWLDTRI AMRVQAERPP WYISVVGGAL 660

YKTVTANTAT VSDYVGYLTK GQIPPKHISL VYTTVALNID GKKYTIDTVR SGHGSYRLRM 720

NGSTVDANVQ ILCDGGLLMQ LDGNSHVIYA EEEASGTRLL IDGKTCMLQN DHDPSKLLAE 780

TPCKLLRFLV ADGAHVDADV PYAEVEVMKM CMPLLSPASG VIHVVMSEGQ AMQAGDLIAR 840

LDLDDPSAVK RAEPFEDTFP QMGLPIAASG QVHKLCAASL NACRMILAGY EHDIDKVVPE 900

LVYCLDTPEL PFLQWEELMS VLATRLPRNL KSELEGKYEE YKVKFDSGII NDFPANMLRV 960

IIEENLACGS EKEKATNERL VEPLMSLLKS YEGGRESHAH FVVKSLFEEY LYVEELFSDG 1020

IQSDVIERLR LQHSKDLQKV VDIVLSHQSV RNKTKLILKL MESLVYPNPA AYRDQLIRFS 1080

SLNHKAYYKL ALKASELLEQ TKLSELRARI ARSLSELEMF TEESKGLSMH KREIAIKESM 1140

EDLVTAPLPV EDALISLFDC SDTTVQQRVI ETYIARLYQP HLVKDSIKMK WIESGVIALW 1200

EFPEGHFDAR NGGAVLGDKR WGAMVIVKSL ESLSMAIRFA LKETSHYTSS EGNMMHIALL 1260

GADNKMHIIQ ESGDDADRIA KLPLILKDNV TDLHASGVKT ISFIVQRDEA RMTMRRTFLW 1320

SDEKLSYEEE PILRHVEPPL SALLELDKLK VKGYNEMKYT PSRDRQWHIY TLRNTENPKM 1380

LHRVFFRTLV RQPSVSNKFS SGQIGDMEVG SAEEPLSFTS TSILRSLMTA IEELELHAIR 1440

TGHSHMYLHV LKEQKLLDLV PVSGNTVLDV GQDEATAYSL LKEMAMKIHE LVGARMHHLS 1500

VCQWEVKLKL DCDGPASGTW RIVTTNVTSH TCTVDIYREM EDKESRKLVY HPATPAAGPL 1560

HGVALNNPYQ PLSVIDLKRC SARNNRTTYC YDFPLAFETA VRKSWSSSTS GASKGVENAQ 1620

CYVKATELVF ADKHGSWGTP LVQMDRPAGL NDIGMVAWTL KMSTPEFPSG REIIVVANDI 1680

TFRAGSFGPR EDAFFEAVTN LACEKKLPLI YLAANSGARI GIADEVKSCF RVGWSDDGSP 1740

ERGFQYIYLS EEDYARIGTS VIAHKMQLDS GEIRWVIDSV VGKEDGLGVE NIHGSAAIAS 1800

AYSRAYKETF TLTFVTGRTV GIGAYLARLG IRCIQRLDQP IILTGYSALN KLLGREVYSS 1860

HMQLGGPKIM ATNGVVHLTV SDDLEGVSNI LRWLSYVPAY IGGPLPVTTP LDPPDRPVAY 1920

IPENSCDPRA AIRGVDDSQG KWLGGMFDKD SFVETFEGWA KTVVTGRAKL GGIPVGVIAV 1980

ETQTMMQTIP ADPGQLDSRE QSVPRAGQVW FPDSATKTAQ ALLDFNREGL PLFILANWRG 2040

FSGGQRDLFE GILQAGSTIV ENLRTYNQPA FVYIPMAAEL RGGAWVVVDS KINPDRIECY 2100

AERTAKGNVL EPQGLIEIKF RSEELQDCMS RLDPTLIDLK AKLEVANKNG SADTKSLQEN 2160

IEARTKQLMP LYTQIAIRFA ELHDTSLRMA AKGVIKKVVD WEESRSFFYK RLRRRISEDV 2220

LAKEIRAVAG EQFSHQPAIE LIKKWYSASH AAEWDDDDAF VAWMDNPENY KDYIQDLKAQ 2280

RVSQSLSSLS DSSSDLQALP QGLSMLLDKM DPSRRAQLVE EIRKVLG 2327

<210> 5

<211> 6984

<212> DNA

<213> mutant nucleotide sequences of rice (Oryza sativa) Jiahua No.1 ACCase-5191 and ACCase-6826

<400> 5

atgacatcca cacatgtggc gacattggga gttggtgccc aggcacctcc tcgtcaccag 60

aaaaagtcag ctggcactgc atttgtatca tctgggtcat caagaccctc ataccgaaag 120

aatggtcagc gtactcggtc acttagggaa gaaagcaatg gaggagtgtc tgattccaaa 180

aagcttaacc actctattcg ccaaggtctt gctggcatca ttgacctccc aaatgacgca 240

gcttcagaag ttgatatttc acatggttcc gaagatccca gggggcctac ggtcccaggt 300

tcctaccaaa tgaatgggat tatcaatgaa acacataatg ggaggcatgc ttcagtctcc 360

aaggttgttg agttttgtac ggcacttggt ggcaaaacac caattcacag tgtattagtg 420

gccaacaatg gaatggcagc agctaagttc atgcggagtg tccgaacatg ggctaatgat 480

acttttggat cagagaaggc aattcagctg atagctatgg caactccgga ggatctgagg 540

ataaatgcag agcacatcag aattgccgat caatttgtag aggtacctgg tggaacaaac 600

aacaacaact atgcaaatgt ccaactcata gtggagatag cagagagaac aggtgtttct 660

gctgtttggc ctggttgggg tcatgcatct gagaatcctg aacttccaga tgcgctgact 720

gcaaaaggaa ttgtttttct tgggccacca gcatcatcaa tgcatgcatt aggagacaag 780

gttggctcag ctctcattgc tcaagcagct ggagttccaa cacttgcttg gagtggatca 840

catgtggaag ttcctctgga gtgttgcttg gactcaatac ctgatgagat gtatagaaaa 900

gcttgtgtta ctaccacaga ggaagcagtt gcaagttgtc aggtggttgg ttatcctgcc 960

atgattaagg catcttgggg tggtggtggt aaaggaataa ggaaggttca taatgatgat 1020

gaggttagga cattatttaa gcaagttcaa ggcgaagtac ctggttcccc aatatttatc 1080

atgaggctag ctgctcagag tcgacatctt gaagttcagt tgctttgtga tcaatatggc 1140

aacgtagcag cacttcacag tcgagattgc agtgtacaac ggcgacacca aaagataatc 1200

gaggaaggac cagttactgt tgctcctcgt gagactgtga aagagcttga gcaggcagca 1260

cggaggcttg ctaaagctgt gggttatgtt ggtgctgcta ctgttgaata cctttacagc 1320

atggaaactg gtgaatatta ttttctggaa cttaatccac ggctacaggt tgagcatcct 1380

gtcactgagt ggatagctga agtaaatttg cctgcggctc aagttgctgt tggaatgggt 1440

ataccccttt ggcagattcc agagatcagg cgcttctacg gaatgaacca tggaggaggc 1500

tatgaccttt ggaggaaaac agcagctcta gcgactccat ttaactttga tgaagtagat 1560

tctaaatggc caaaaggcca ctgcgtagct gttagaataa ctagcgagga tccagatgat 1620

gggtttaagc ctactggtgg aaaagtaaag gagataagtt tcaagagtaa accaaatgtt 1680

tgggcctatt tctcagtaaa gtctggtgga ggcatccatg aattcgctga ttctcagttc 1740

ggacatgttt ttgcgtatgg aactactaga tcggcagcaa taactaccat ggctcttgca 1800

ctaaaagagg ttcaaattcg tggagaaatt cattcaaacg tagactacac agttgaccta 1860

ttaaatgcct cagattttag agaaaataag attcatactg gttggctgga taccaggata 1920

gccatgcgtg ttcaagctga gaggcctcca tggtatattt cagtcgttgg aggggcttta 1980

tataaaacag taactgccaa cacggccact gtttctgatt atgttggtta tcttaccaag 2040

ggccagattc caccaaagca tatatccctt gtctatacga ctgttgcttt gaatatagat 2100

gggaaaaaat atacaatcga tactgtgagg agtggacatg gtagctacag attgcgaatg 2160

aatggatcaa cggttgacgc aaatgtacaa atattatgtg atggtgggct tttaatgcag 2220

ctggatggaa acagccatgt aatttatgct gaagaagagg ccagtggtac acgacttctt 2280

attgatggaa agacatgcat gttacagaat gaccatgacc catcaaagtt attagctgag 2340

acaccatgca aacttcttcg tttcttggtt gctgatggtg ctcatgttga tgctgatgta 2400

ccatatgcgg aagttgaggt tatgaagatg tgcatgcccc tcttatcacc cgcttctggt 2460

gtcatacatg ttgtaatgtc tgagggccaa gcaatgcagg ctggtgatct tatagctagg 2520

ctggatcttg atgacccttc tgctgttaag agagctgagc cgttcgaaga tacttttcca 2580

caaatgggtc tccctattgc tgcttctggc caagttcaca aattatgtgc tgcaagtctg 2640

aatgcttgtc gaatgatcct tgcggggtat gagcatgata ttgacaaggt tgtgccagag 2700

ttggtatact gcctagacac tccggagctt cctttcctgc agtgggagga gcttatgtct 2760

gttttagcaa ctagacttcc aagaaatctt aaaagtgagt tggagggcaa atatgaggaa 2820

tacaaagtaa aatttgactc tgggataatc aatgatttcc ctgccaatat gctacgagtg 2880

ataattgagg aaaatcttgc atgtggttct gagaaggaga aggctacaaa tgagaggctt 2940

gttgagcctc ttatgagcct actgaagtca tatgagggtg ggagagaaag tcatgctcac 3000

tttgttgtca agtccctttt tgaggagtat ctctatgttg aagaattgtt cagtgatgga 3060

attcagtctg atgtgattga gcgtctgcgc cttcaacata gtaaagacct acagaaggtc 3120

gtagacattg tgttgtccca ccagagtgtt agaaataaaa ctaagctgat actaaaactc 3180

atggagagtc tggtctatcc aaatcctgct gcctacaggg atcaattgat tcgcttttct 3240

tcccttaatc acaaagcgta ttacaagttg gcacttaaag ctagtgaact tcttgaacaa 3300

acaaaactta gtgagctccg tgcaagaata gcaaggagcc tttcagagct ggagatgttt 3360

actgaggaaa gcaagggtct ctccatgcat aagcgagaaa ttgccattaa ggagagcatg 3420

gaagatttag tcactgctcc actgccagtt gaagatgcgc tcatttcttt atttgattgt 3480

agtgatacaa ctgttcaaca gagagtgatt gagacttata tagctcgatt ataccagcct 3540

catcttgtaa aggacagtat caaaatgaaa tggatagaat cgggtgttat tgctttatgg 3600

gaatttcctg aagggcattt tgatgcaaga aatggaggag cggttcttgg tgacaaaaga 3660

tggggtgcca tggtcattgt caagtctctt gaatcacttt caatggccat tagatttgca 3720

ctaaaggaga catcacacta cactagctct gagggcaata tgatgcatat tgctttgttg 3780

ggtgctgata ataagatgca tataattcaa gaaagtggtg atgatgctga cagaatagcc 3840

aaacttccct tgatactaaa ggataatgta accgatctgc atgcctctgg tgtgaaaaca 3900

ataagtttca ttgttcaaag agatgaagca cggatgacaa tgcgtcgtac cttcctttgg 3960

tctgatgaaa agctttctta tgaggaagag ccaattctcc ggcatgtgga acctcctctt 4020

tctgcacttc ttgagttgga caagttgaaa gtgaaaggat acaatgaaat gaagtatacc 4080

ccatcacggg atcgtcaatg gcatatctac acacttagaa atactgaaaa ccccaaaatg 4140

ttgcaccggg tatttttccg aacccttgtc aggcaaccca gtgtatccaa caagttttct 4200

tcgggccaga ttggtgacat ggaagttggg agtgctgaag aacctctgtc atttacatca 4260

accagcatat taagatcttt gatgactgct atagaggaat tggagcttca cgcaattaga 4320

actggccatt cacacatgta tttgcatgta ttgaaagaac aaaagcttct tgatcttgtt 4380

ccagtttcag ggaatacagt tttggatgtt ggtcaagatg aagctactgc atattcactt 4440

ttaaaagaaa tggctatgaa gatacatgaa cttgttggtg caagaatgca ccatctttct 4500

gtatgccaat gggaagtgaa acttaagttg gactgcgatg gtcctgccag tggtacctgg 4560

aggattgtaa caaccaatgt tactagtcac acttgcactg tggatatcta ccgtgagatg 4620

gaagataaag aatcacggaa gttagtatac catcccgcca ctccggcggc tggtcctctg 4680

catggtgtgg cactgaataa tccatatcag cctttgagtg tcattgatct caaacgctgt 4740

tctgctagga ataatagaac tacatactgc tatgattttc cactggcatt tgaaactgca 4800

gtgaggaagt catggtcctc tagtacctct ggtgcttcta aaggtgttga aaatgcccaa 4860

tgttatgtta aagctacaga gttggtattt gcggacaaac atgggtcatg gggcactcct 4920

ttagttcaaa tggaccggcc tgctgggctc aatgacattg gtatggtagc ttggaccttg 4980

aagatgtcca ctcctgaatt tcctagtggt agggagatta ttgttgttgc aaatgatatt 5040

acgttcagag ctggatcatt tggcccaagg gaagatgcat tttttgaagc tgttaccaac 5100

ctagcctgtg agaagaaact tcctcttatt tatttggcag caaattctgg tgctcgaatt 5160

ggcatagcag atgaagtgaa atcttgcttc tgtgttgggt ggtctgatga tggcagccct 5220

gaacgtgggt ttcagtacat ttatctaagc gaagaagact atgctcgtat tggcacttct 5280

gtcatagcac ataagatgca gctagacagt ggtgaaatta ggtgggttat tgattctgtt 5340

gtgggcaagg aagatggact tggtgtggag aatatacatg gaagtgctgc tattgccagt 5400

gcttattcta gggcatataa ggagacattt acacttacat ttgtgactgg aagaactgtt 5460

ggaataggag cttatcttgc tcgacttggc atccggtgca tacagcgtct tgaccagcct 5520

attattctta caggctattc tgcactgaac aagcttcttg ggcgggaagt gtacagctcc 5580

cacatgcagt tgggtggtcc caaaatcatg gcaactaatg gtgttgtcca tcttactgtt 5640

tcagatgacc ttgaaggcgt ttctaatata ttgaggtggc tcagttatgt tcctgcctac 5700

attggtggac cacttccagt aacaacaccg ttggacccac cggacagacc tgttgcatac 5760

attcctgaga actcgtgtga tcctcgagcg gctatccgtg gtgttgatga cagccaaggg 5820

aaatggttag gtggtatgtt tgataaagac agctttgtgg aaacatttga aggttgggct 5880

aagacagtgg ttactggcag agcaaagctt ggtggaattc cagtgggtgt gatagctgtg 5940

gagactcaga ccatgatgca aactatccct gctgaccctg gtcagcttga ttcccgtgag 6000

caatctgttc ctcgtgctgg acaagtgtgg tttccagatt ctgcaaccaa gactgcgcag 6060

gcattgctgg acttcaaccg tgaaggatta cctctgttca tcctcgctaa ctggagaggc 6120

ttctctggtg gacaaagaga tctttttgaa ggaattcttc aggctggctc gactattgtt 6180

gagaacctta ggacatacaa tcagcctgcc tttgtctaca ttcccatggc tgcagagcta 6240

cgaggagggg cttgggttgt ggttgatagc aagataaacc cagaccgcat tgagtgctat 6300

gctgagagga ctgcaaaagg caatgttctg gaaccgcaag ggttaattga gatcaagttc 6360

aggtcagagg aactccagga ttgcatgagt cggcttgacc caacattaat tgatctgaaa 6420

gcaaaactcg aagtagcaaa taaaaatgga agtgctgaca caaaatcgct tcaagaaaat 6480

atagaagctc gaacaaaaca gttgatgcct ctatatactc agattgcgat acggtttgct 6540

gaattgcatg atacatccct cagaatggct gcgaaaggtg tgattaagaa agttgtggac 6600

tgggaagaat cacgatcttt cttctataag agattacgga ggaggatctc tgaggatgtt 6660

cttgcaaaag aaattagagc tgtagcaggt gagcagtttt cccaccaacc agcaatcgag 6720

ctgatcaaga aatggtattc agcttcacat gcagctgaat gggatgatga cgatgctttt 6780

gttgcttgga tggataaccc tgaaaactac aaggattata ttcaagatct taaggctcaa 6840

agagtatccc aatccctctc aagtctttca gattccagct cagatttgca agccctgcca 6900

cagggtcttt ccatgttact agataagatg gatccctcta gaagagctca acttgttgaa 6960

gaaatcagga aggtccttgg ttga 6984

<210> 6

<211> 2327

<212> PRT

<213> mutant amino acid sequences of rice (Oryza sativa) Jiahua No.1 ACCase-1731 and ACCase-5191

<400> 6

MTSTHVATLG VGAQAPPRHQ KKSAGTAFVS SGSSRPSYRK NGQRTRSLRE ESNGGVSDSK 60

KLNHSIRQGL AGIIDLPNDA ASEVDISHGS EDPRGPTVPG SYQMNGIINE THNGRHASVS 120

KVVEFCTALG GKTPIHSVLV ANNGMAAAKF MRSVRTWAND TFGSEKAIQL IAMATPEDLR 180

INAEHIRIAD QFVEVPGGTN NNNYANVQLI VEIAERTGVS AVWPGWGHAS ENPELPDALT 240

AKGIVFLGPP ASSMHALGDK VGSALIAQAA GVPTLAWSGS HVEVPLECCL DSIPDEMYRK 300

ACVTTTEEAV ASCQVVGYPA MIKASWGGGG KGIRKVHNDD EVRTLFKQVQ GEVPGSPIFI 360

MRLAAQSRHL EVQLLCDQYG NVAALHSRDC SVQRRHQKII EEGPVTVAPR ETVKELEQAA 420

RRLAKAVGYV GAATVEYLYS METGEYYFLE LNPRLQVEHP VTEWIAEVNL PAAQVAVGMG 480

IPLWQIPEIR RFYGMNHGGG YDLWRKTAAL ATPFNFDEVD SKWPKGHCVA VRITSEDPDD 540

GFKPTGGKVK EISFKSKPNV WAYFSVKSGG GIHEFADSQF GHVFAYGTTR SAAITTMALA 600

LKEVQIRGEI HSNVDYTVDL LNASDFRENK IHTGWLDTRI AMRVQAERPP WYISVVGGAL 660

YKTVTANTAT VSDYVGYLTK GQIPPKHISL VYTTVALNID GKKYTIDTVR SGHGSYRLRM 720

NGSTVDANVQ ILCDGGLLMQ LDGNSHVIYA EEEASGTRLL IDGKTCMLQN DHDPSKLLAE 780

TPCKLLRFLV ADGAHVDADV PYAEVEVMKM CMPLLSPASG VIHVVMSEGQ AMQAGDLIAR 840

LDLDDPSAVK RAEPFEDTFP QMGLPIAASG QVHKLCAASL NACRMILAGY EHDIDKVVPE 900

LVYCLDTPEL PFLQWEELMS VLATRLPRNL KSELEGKYEE YKVKFDSGII NDFPANMLRV 960

IIEENLACGS EKEKATNERL VEPLMSLLKS YEGGRESHAH FVVKSLFEEY LYVEELFSDG 1020

IQSDVIERLR LQHSKDLQKV VDIVLSHQSV RNKTKLILKL MESLVYPNPA AYRDQLIRFS 1080

SLNHKAYYKL ALKASELLEQ TKLSELRARI ARSLSELEMF TEESKGLSMH KREIAIKESM 1140

EDLVTAPLPV EDALISLFDC SDTTVQQRVI ETYIARLYQP HLVKDSIKMK WIESGVIALW 1200

EFPEGHFDAR NGGAVLGDKR WGAMVIVKSL ESLSMAIRFA LKETSHYTSS EGNMMHIALL 1260

GADNKMHIIQ ESGDDADRIA KLPLILKDNV TDLHASGVKT ISFIVQRDEA RMTMRRTFLW 1320

SDEKLSYEEE PILRHVEPPL SALLELDKLK VKGYNEMKYT PSRDRQWHIY TLRNTENPKM 1380

LHRVFFRTLV RQPSVSNKFS SGQIGDMEVG SAEEPLSFTS TSILRSLMTA IEELELHAIR 1440

TGHSHMYLHV LKEQKLLDLV PVSGNTVLDV GQDEATAYSL LKEMAMKIHE LVGARMHHLS 1500

VCQWEVKLKL DCDGPASGTW RIVTTNVTSH TCTVDIYREM EDKESRKLVY HPATPAAGPL 1560

HGVALNNPYQ PLSVIDLKRC SARNNRTTYC YDFPLAFETA VRKSWSSSTS GASKGVENAQ 1620

CYVKATELVF ADKHGSWGTP LVQMDRPAGL NDIGMVAWTL KMSTPEFPSG REIIVVANDI 1680

TFRAGSFGPR EDAFFEAVTN LACEKKLPLI YLAANSGARI GIADEVKSCF CVGWSDDGSP 1740

ERGFQYIYLS EEDYARIGTS VIAHKMQLDS GEIRWVIDSV VGKEDGLGVE NIHGSAAIAS 1800

AYSRAYKETF TLTFVTGRTV GIGAYLARLG IRCIQRLDQP IILTGYSALN KLLGREVYSS 1860

HMQLGGPKIM ATNGVVHLTV SDDLEGVSNI LRWLSYVPAY IGGPLPVTTP LDPPDRPVAY 1920

IPENSCDPRA AIRGVDDSQG KWLGGMFDKD SFVETFEGWA KTVVTGRAKL GGIPVGVIAV 1980

ETQTMMQTIP ADPGQLDSRE QSVPRAGQVW FPDSATKTAQ ALLDFNREGL PLFILANWRG 2040

FSGGQRDLFE GILQAGSTIV ENLRTYNQPA FVYIPMAAEL RGGAWVVVDS KINPDRIECY 2100

AERTAKGNVL EPQGLIEIKF RSEELQDCMS RLDPTLIDLK AKLEVANKNG SADTKSLQEN 2160

IEARTKQLMP LYTQIAIRFA ELHDTSLRMA AKGVIKKVVD WEESRSFFYK RLRRRISEDV 2220

LAKEIRAVAG EQFSHQPAIE LIKKWYSASH AAEWDDDDAF VAWMDNPENY KDYIQDLKAQ 2280

RVSQSLSSLS DSSSDLQALP QGLSMLLDKM DPSRRAQLVE EIRKVLG 2327

<210> 7

<211> 6984

<212> DNA

<213> wild type Gene sequence of ACCase No.1 of Rice (Oryza sativa) Jiahua

<400> 7

atgacatcca cacatgtggc gacattggga gttggtgccc aggcacctcc tcgtcaccag 60

aaaaagtcag ctggcactgc atttgtatca tctgggtcat caagaccctc ataccgaaag 120

aatggtcagc gtactcggtc acttagggaa gaaagcaatg gaggagtgtc tgattccaaa 180

aagcttaacc actctattcg ccaaggtctt gctggcatca ttgacctccc aaatgacgca 240

gcttcagaag ttgatatttc acatggttcc gaagatccca gggggcctac ggtcccaggt 300

tcctaccaaa tgaatgggat tatcaatgaa acacataatg ggaggcatgc ttcagtctcc 360

aaggttgttg agttttgtac ggcacttggt ggcaaaacac caattcacag tgtattagtg 420

gccaacaatg gaatggcagc agctaagttc atgcggagtg tccgaacatg ggctaatgat 480

acttttggat cagagaaggc aattcagctg atagctatgg caactccgga ggatctgagg 540

ataaatgcag agcacatcag aattgccgat caatttgtag aggtacctgg tggaacaaac 600

aacaacaact atgcaaatgt ccaactcata gtggagatag cagagagaac aggtgtttct 660

gctgtttggc ctggttgggg tcatgcatct gagaatcctg aacttccaga tgcgctgact 720

gcaaaaggaa ttgtttttct tgggccacca gcatcatcaa tgcatgcatt aggagacaag 780

gttggctcag ctctcattgc tcaagcagct ggagttccaa cacttgcttg gagtggatca 840

catgtggaag ttcctctgga gtgttgcttg gactcaatac ctgatgagat gtatagaaaa 900

gcttgtgtta ctaccacaga ggaagcagtt gcaagttgtc aggtggttgg ttatcctgcc 960

atgattaagg catcttgggg tggtggtggt aaaggaataa ggaaggttca taatgatgat 1020

gaggttagga cattatttaa gcaagttcaa ggcgaagtac ctggttcccc aatatttatc 1080

atgaggctag ctgctcagag tcgacatctt gaagttcagt tgctttgtga tcaatatggc 1140

aacgtagcag cacttcacag tcgagattgc agtgtacaac ggcgacacca aaagataatc 1200

gaggaaggac cagttactgt tgctcctcgt gagactgtga aagagcttga gcaggcagca 1260

cggaggcttg ctaaagctgt gggttatgtt ggtgctgcta ctgttgaata cctttacagc 1320

atggaaactg gtgaatatta ttttctggaa cttaatccac ggctacaggt tgagcatcct 1380

gtcactgagt ggatagctga agtaaatttg cctgcggctc aagttgctgt tggaatgggt 1440

ataccccttt ggcagattcc agagatcagg cgcttctacg gaatgaacca tggaggaggc 1500

tatgaccttt ggaggaaaac agcagctcta gcgactccat ttaactttga tgaagtagat 1560

tctaaatggc caaaaggcca ctgcgtagct gttagaataa ctagcgagga tccagatgat 1620

gggtttaagc ctactggtgg aaaagtaaag gagataagtt tcaagagtaa accaaatgtt 1680

tgggcctatt tctcagtaaa gtctggtgga ggcatccatg aattcgctga ttctcagttc 1740

ggacatgttt ttgcgtatgg aactactaga tcggcagcaa taactaccat ggctcttgca 1800

ctaaaagagg ttcaaattcg tggagaaatt cattcaaacg tagactacac agttgaccta 1860

ttaaatgcct cagattttag agaaaataag attcatactg gttggctgga taccaggata 1920

gccatgcgtg ttcaagctga gaggcctcca tggtatattt cagtcgttgg aggggcttta 1980

tataaaacag taactgccaa cacggccact gtttctgatt atgttggtta tcttaccaag 2040

ggccagattc caccaaagca tatatccctt gtctatacga ctgttgcttt gaatatagat 2100

gggaaaaaat atacaatcga tactgtgagg agtggacatg gtagctacag attgcgaatg 2160

aatggatcaa cggttgacgc aaatgtacaa atattatgtg atggtgggct tttaatgcag 2220

ctggatggaa acagccatgt aatttatgct gaagaagagg ccagtggtac acgacttctt 2280

attgatggaa agacatgcat gttacagaat gaccatgacc catcaaagtt attagctgag 2340

acaccatgca aacttcttcg tttcttggtt gctgatggtg ctcatgttga tgctgatgta 2400

ccatatgcgg aagttgaggt tatgaagatg tgcatgcccc tcttatcacc cgcttctggt 2460

gtcatacatg ttgtaatgtc tgagggccaa gcaatgcagg ctggtgatct tatagctagg 2520

ctggatcttg atgacccttc tgctgttaag agagctgagc cgttcgaaga tacttttcca 2580

caaatgggtc tccctattgc tgcttctggc caagttcaca aattatgtgc tgcaagtctg 2640

aatgcttgtc gaatgatcct tgcggggtat gagcatgata ttgacaaggt tgtgccagag 2700

ttggtatact gcctagacac tccggagctt cctttcctgc agtgggagga gcttatgtct 2760

gttttagcaa ctagacttcc aagaaatctt aaaagtgagt tggagggcaa atatgaggaa 2820

tacaaagtaa aatttgactc tgggataatc aatgatttcc ctgccaatat gctacgagtg 2880

ataattgagg aaaatcttgc atgtggttct gagaaggaga aggctacaaa tgagaggctt 2940

gttgagcctc ttatgagcct actgaagtca tatgagggtg ggagagaaag tcatgctcac 3000

tttgttgtca agtccctttt tgaggagtat ctctatgttg aagaattgtt cagtgatgga 3060

attcagtctg atgtgattga gcgtctgcgc cttcaacata gtaaagacct acagaaggtc 3120

gtagacattg tgttgtccca ccagagtgtt agaaataaaa ctaagctgat actaaaactc 3180

atggagagtc tggtctatcc aaatcctgct gcctacaggg atcaattgat tcgcttttct 3240

tcccttaatc acaaagcgta ttacaagttg gcacttaaag ctagtgaact tcttgaacaa 3300

acaaaactta gtgagctccg tgcaagaata gcaaggagcc tttcagagct ggagatgttt 3360

actgaggaaa gcaagggtct ctccatgcat aagcgagaaa ttgccattaa ggagagcatg 3420

gaagatttag tcactgctcc actgccagtt gaagatgcgc tcatttcttt atttgattgt 3480

agtgatacaa ctgttcaaca gagagtgatt gagacttata tagctcgatt ataccagcct 3540

catcttgtaa aggacagtat caaaatgaaa tggatagaat cgggtgttat tgctttatgg 3600

gaatttcctg aagggcattt tgatgcaaga aatggaggag cggttcttgg tgacaaaaga 3660

tggggtgcca tggtcattgt caagtctctt gaatcacttt caatggccat tagatttgca 3720

ctaaaggaga catcacacta cactagctct gagggcaata tgatgcatat tgctttgttg 3780

ggtgctgata ataagatgca tataattcaa gaaagtggtg atgatgctga cagaatagcc 3840

aaacttccct tgatactaaa ggataatgta accgatctgc atgcctctgg tgtgaaaaca 3900

ataagtttca ttgttcaaag agatgaagca cggatgacaa tgcgtcgtac cttcctttgg 3960

tctgatgaaa agctttctta tgaggaagag ccaattctcc ggcatgtgga acctcctctt 4020

tctgcacttc ttgagttgga caagttgaaa gtgaaaggat acaatgaaat gaagtatacc 4080

ccatcacggg atcgtcaatg gcatatctac acacttagaa atactgaaaa ccccaaaatg 4140

ttgcaccggg tatttttccg aacccttgtc aggcaaccca gtgtatccaa caagttttct 4200

tcgggccaga ttggtgacat ggaagttggg agtgctgaag aacctctgtc atttacatca 4260

accagcatat taagatcttt gatgactgct atagaggaat tggagcttca cgcaattaga 4320

actggccatt cacacatgta tttgcatgta ttgaaagaac aaaagcttct tgatcttgtt 4380

ccagtttcag ggaatacagt tttggatgtt ggtcaagatg aagctactgc atattcactt 4440

ttaaaagaaa tggctatgaa gatacatgaa cttgttggtg caagaatgca ccatctttct 4500

gtatgccaat gggaagtgaa acttaagttg gactgcgatg gtcctgccag tggtacctgg 4560

aggattgtaa caaccaatgt tactagtcac acttgcactg tggatatcta ccgtgagatg 4620

gaagataaag aatcacggaa gttagtatac catcccgcca ctccggcggc tggtcctctg 4680

catggtgtgg cactgaataa tccatatcag cctttgagtg tcattgatct caaacgctgt 4740

tctgctagga ataatagaac tacatactgc tatgattttc cactggcatt tgaaactgca 4800

gtgaggaagt catggtcctc tagtacctct ggtgcttcta aaggtgttga aaatgcccaa 4860

tgttatgtta aagctacaga gttggtattt gcggacaaac atgggtcatg gggcactcct 4920

ttagttcaaa tggaccggcc tgctgggctc aatgacattg gtatggtagc ttggaccttg 4980

aagatgtcca ctcctgaatt tcctagtggt agggagatta ttgttgttgc aaatgatatt 5040

acgttcagag ctggatcatt tggcccaagg gaagatgcat tttttgaagc tgttaccaac 5100

ctagcctgtg agaagaaact tcctcttatt tatttggcag caaattctgg tgctcgaatt 5160

ggcatagcag atgaagtgaa atcttgcttc cgtgttgggt ggtctgatga tggcagccct 5220

gaacgtgggt ttcagtacat ttatctaagc gaagaagact atgctcgtat tggcacttct 5280

gtcatagcac ataagatgca gctagacagt ggtgaaatta ggtgggttat tgattctgtt 5340

gtgggcaagg aagatggact tggtgtggag aatatacatg gaagtgctgc tattgccagt 5400

gcttattcta gggcatataa ggagacattt acacttacat ttgtgactgg aagaactgtt 5460

ggaataggag cttatcttgc tcgacttggc atccggtgca tacagcgtct tgaccagcct 5520

attattctta caggctattc tgcactgaac aagcttcttg ggcgggaagt gtacagctcc 5580

cacatgcagt tgggtggtcc caaaatcatg gcaactaatg gtgttgtcca tcttactgtt 5640

tcagatgacc ttgaaggcgt ttctaatata ttgaggtggc tcagttatgt tcctgcctac 5700

attggtggac cacttccagt aacaacaccg ttggacccac cggacagacc tgttgcatac 5760

attcctgaga actcgtgtga tcctcgagcg gctatccgtg gtgttgatga cagccaaggg 5820

aaatggttag gtggtatgtt tgataaagac agctttgtgg aaacatttga aggttgggct 5880

aagacagtgg ttactggcag agcaaagctt ggtggaattc cagtgggtgt gatagctgtg 5940

gagactcaga ccatgatgca aactatccct gctgaccctg gtcagcttga ttcccgtgag 6000

caatctgttc ctcgtgctgg acaagtgtgg tttccagatt ctgcaaccaa gactgcgcag 6060

gcattgctgg acttcaaccg tgaaggatta cctctgttca tcctcgctaa ctggagaggc 6120

ttctctggtg gacaaagaga tctttttgaa ggaattcttc aggctggctc gactattgtt 6180

gagaacctta ggacatacaa tcagcctgcc tttgtctaca ttcccatggc tgcagagcta 6240

cgaggagggg cttgggttgt ggttgatagc aagataaacc cagaccgcat tgagtgctat 6300

gctgagagga ctgcaaaagg caatgttctg gaaccgcaag ggttaattga gatcaagttc 6360

aggtcagagg aactccagga ttgcatgagt cggcttgacc caacattaat tgatctgaaa 6420

gcaaaactcg aagtagcaaa taaaaatgga agtgctgaca caaaatcgct tcaagaaaat 6480

atagaagctc gaacaaaaca gttgatgcct ctatatactc agattgcgat acggtttgct 6540

gaattgcatg atacatccct cagaatggct gcgaaaggtg tgattaagaa agttgtggac 6600

tgggaagaat cacgatcttt cttctataag agattacgga ggaggatctc tgaggatgtt 6660

cttgcaaaag aaattagagc tgtagcaggt gagcagtttt cccaccaacc agcaatcgag 6720

ctgatcaaga aatggtattc agcttcacat gcagctgaat gggatgatga cgatgctttt 6780

gttgcttgga tggataaccc tgaaaactac aaggattata ttcaatatct taaggctcaa 6840

agagtatccc aatccctctc aagtctttca gattccagct cagatttgca agccctgcca 6900

cagggtcttt ccatgttact agataagatg gatccctcta gaagagctca acttgttgaa 6960

gaaatcagga aggtccttgg ttga 6984

<210> 8

<211> 2327

<212> PRT

<213> wild type amino acid sequence of ACCase No.1 of rice (Oryza sativa) Jiahua

<400> 8

Met Thr Ser Thr His Val Ala Thr Leu Gly Val Gly Ala Gln Ala Pro

1 5 10 15

Pro Arg His Gln Lys Lys Ser Ala Gly Thr Ala Phe Val Ser Ser Gly

20 25 30

Ser Ser Arg Pro Ser Tyr Arg Lys Asn Gly Gln Arg Thr Arg Ser Leu

35 40 45

Arg Glu Glu Ser Asn Gly Gly Val Ser Asp Ser Lys Lys Leu Asn His

50 55 60

Ser Ile Arg Gln Gly Leu Ala Gly Ile Ile Asp Leu Pro Asn Asp Ala

65 70 75 80

Ala Ser Glu Val Asp Ile Ser His Gly Ser Glu Asp Pro Arg Gly Pro

85 90 95

Thr Val Pro Gly Ser Tyr Gln Met Asn Gly Ile Ile Asn Glu Thr His

100 105 110

Asn Gly Arg His Ala Ser Val Ser Lys Val Val Glu Phe Cys Thr Ala

115 120 125

Leu Gly Gly Lys Thr Pro Ile His Ser Val Leu Val Ala Asn Asn Gly

130 135 140

Met Ala Ala Ala Lys Phe Met Arg Ser Val Arg Thr Trp Ala Asn Asp

145 150 155 160

Thr Phe Gly Ser Glu Lys Ala Ile Gln Leu Ile Ala Met Ala Thr Pro

165 170 175

Glu Asp Leu Arg Ile Asn Ala Glu His Ile Arg Ile Ala Asp Gln Phe

180 185 190

Val Glu Val Pro Gly Gly Thr Asn Asn Asn Asn Tyr Ala Asn Val Gln

195 200 205

Leu Ile Val Glu Ile Ala Glu Arg Thr Gly Val Ser Ala Val Trp Pro

210 215 220

Gly Trp Gly His Ala Ser Glu Asn Pro Glu Leu Pro Asp Ala Leu Thr

225 230 235 240

Ala Lys Gly Ile Val Phe Leu Gly Pro Pro Ala Ser Ser Met His Ala

245 250 255

Leu Gly Asp Lys Val Gly Ser Ala Leu Ile Ala Gln Ala Ala Gly Val

260 265 270

Pro Thr Leu Ala Trp Ser Gly Ser His Val Glu Val Pro Leu Glu Cys

275 280 285

Cys Leu Asp Ser Ile Pro Asp Glu Met Tyr Arg Lys Ala Cys Val Thr

290 295 300

Thr Thr Glu Glu Ala Val Ala Ser Cys Gln Val Val Gly Tyr Pro Ala

305 310 315 320

Met Ile Lys Ala Ser Trp Gly Gly Gly Gly Lys Gly Ile Arg Lys Val

325 330 335

His Asn Asp Asp Glu Val Arg Thr Leu Phe Lys Gln Val Gln Gly Glu

340 345 350

Val Pro Gly Ser Pro Ile Phe Ile Met Arg Leu Ala Ala Gln Ser Arg

355 360 365

His Leu Glu Val Gln Leu Leu Cys Asp Gln Tyr Gly Asn Val Ala Ala

370 375 380

Leu His Ser Arg Asp Cys Ser Val Gln Arg Arg His Gln Lys Ile Ile

385 390 395 400

Glu Glu Gly Pro Val Thr Val Ala Pro Arg Glu Thr Val Lys Glu Leu

405 410 415

Glu Gln Ala Ala Arg Arg Leu Ala Lys Ala Val Gly Tyr Val Gly Ala

420 425 430

Ala Thr Val Glu Tyr Leu Tyr Ser Met Glu Thr Gly Glu Tyr Tyr Phe

435 440 445

Leu Glu Leu Asn Pro Arg Leu Gln Val Glu His Pro Val Thr Glu Trp

450 455 460

Ile Ala Glu Val Asn Leu Pro Ala Ala Gln Val Ala Val Gly Met Gly

465 470 475 480

Ile Pro Leu Trp Gln Ile Pro Glu Ile Arg Arg Phe Tyr Gly Met Asn

485 490 495

His Gly Gly Gly Tyr Asp Leu Trp Arg Lys Thr Ala Ala Leu Ala Thr

500 505 510

Pro Phe Asn Phe Asp Glu Val Asp Ser Lys Trp Pro Lys Gly His Cys

515 520 525

Val Ala Val Arg Ile Thr Ser Glu Asp Pro Asp Asp Gly Phe Lys Pro

530 535 540

Thr Gly Gly Lys Val Lys Glu Ile Ser Phe Lys Ser Lys Pro Asn Val

545 550 555 560

Trp Ala Tyr Phe Ser Val Lys Ser Gly Gly Gly Ile His Glu Phe Ala

565 570 575

Asp Ser Gln Phe Gly His Val Phe Ala Tyr Gly Thr Thr Arg Ser Ala

580 585 590

Ala Ile Thr Thr Met Ala Leu Ala Leu Lys Glu Val Gln Ile Arg Gly

595 600 605

Glu Ile His Ser Asn Val Asp Tyr Thr Val Asp Leu Leu Asn Ala Ser

610 615 620

Asp Phe Arg Glu Asn Lys Ile His Thr Gly Trp Leu Asp Thr Arg Ile

625 630 635 640

Ala Met Arg Val Gln Ala Glu Arg Pro Pro Trp Tyr Ile Ser Val Val

645 650 655

Gly Gly Ala Leu Tyr Lys Thr Val Thr Ala Asn Thr Ala Thr Val Ser

660 665 670

Asp Tyr Val Gly Tyr Leu Thr Lys Gly Gln Ile Pro Pro Lys His Ile

675 680 685

Ser Leu Val Tyr Thr Thr Val Ala Leu Asn Ile Asp Gly Lys Lys Tyr

690 695 700

Thr Ile Asp Thr Val Arg Ser Gly His Gly Ser Tyr Arg Leu Arg Met

705 710 715 720

Asn Gly Ser Thr Val Asp Ala Asn Val Gln Ile Leu Cys Asp Gly Gly

725 730 735

Leu Leu Met Gln Leu Asp Gly Asn Ser His Val Ile Tyr Ala Glu Glu

740 745 750

Glu Ala Ser Gly Thr Arg Leu Leu Ile Asp Gly Lys Thr Cys Met Leu

755 760 765

Gln Asn Asp His Asp Pro Ser Lys Leu Leu Ala Glu Thr Pro Cys Lys

770 775 780

Leu Leu Arg Phe Leu Val Ala Asp Gly Ala His Val Asp Ala Asp Val

785 790 795 800

Pro Tyr Ala Glu Val Glu Val Met Lys Met Cys Met Pro Leu Leu Ser

805 810 815

Pro Ala Ser Gly Val Ile His Val Val Met Ser Glu Gly Gln Ala Met

820 825 830

Gln Ala Gly Asp Leu Ile Ala Arg Leu Asp Leu Asp Asp Pro Ser Ala

835 840 845

Val Lys Arg Ala Glu Pro Phe Glu Asp Thr Phe Pro Gln Met Gly Leu

850 855 860

Pro Ile Ala Ala Ser Gly Gln Val His Lys Leu Cys Ala Ala Ser Leu

865 870 875 880

Asn Ala Cys Arg Met Ile Leu Ala Gly Tyr Glu His Asp Ile Asp Lys

885 890 895

Val Val Pro Glu Leu Val Tyr Cys Leu Asp Thr Pro Glu Leu Pro Phe

900 905 910

Leu Gln Trp Glu Glu Leu Met Ser Val Leu Ala Thr Arg Leu Pro Arg

915 920 925

Asn Leu Lys Ser Glu Leu Glu Gly Lys Tyr Glu Glu Tyr Lys Val Lys

930 935 940

Phe Asp Ser Gly Ile Ile Asn Asp Phe Pro Ala Asn Met Leu Arg Val

945 950 955 960

Ile Ile Glu Glu Asn Leu Ala Cys Gly Ser Glu Lys Glu Lys Ala Thr

965 970 975

Asn Glu Arg Leu Val Glu Pro Leu Met Ser Leu Leu Lys Ser Tyr Glu

980 985 990

Gly Gly Arg Glu Ser His Ala His Phe Val Val Lys Ser Leu Phe Glu

995 1000 1005

Glu Tyr Leu Tyr Val Glu Glu Leu Phe Ser Asp Gly Ile Gln Ser Asp

1010 1015 1020

Val Ile Glu Arg Leu Arg Leu Gln His Ser Lys Asp Leu Gln Lys Val

1025 1030 1035 1040

Val Asp Ile Val Leu Ser His Gln Ser Val Arg Asn Lys Thr Lys Leu

1045 1050 1055

Ile Leu Lys Leu Met Glu Ser Leu Val Tyr Pro Asn Pro Ala Ala Tyr

1060 1065 1070

Arg Asp Gln Leu Ile Arg Phe Ser Ser Leu Asn His Lys Ala Tyr Tyr

1075 1080 1085

Lys Leu Ala Leu Lys Ala Ser Glu Leu Leu Glu Gln Thr Lys Leu Ser

1090 1095 1100

Glu Leu Arg Ala Arg Ile Ala Arg Ser Leu Ser Glu Leu Glu Met Phe

1105 1110 1115 1120

Thr Glu Glu Ser Lys Gly Leu Ser Met His Lys Arg Glu Ile Ala Ile

1125 1130 1135

Lys Glu Ser Met Glu Asp Leu Val Thr Ala Pro Leu Pro Val Glu Asp

1140 1145 1150

Ala Leu Ile Ser Leu Phe Asp Cys Ser Asp Thr Thr Val Gln Gln Arg

1155 1160 1165

Val Ile Glu Thr Tyr Ile Ala Arg Leu Tyr Gln Pro His Leu Val Lys

1170 1175 1180

Asp Ser Ile Lys Met Lys Trp Ile Glu Ser Gly Val Ile Ala Leu Trp

1185 1190 1195 1200

Glu Phe Pro Glu Gly His Phe Asp Ala Arg Asn Gly Gly Ala Val Leu

1205 1210 1215

Gly Asp Lys Arg Trp Gly Ala Met Val Ile Val Lys Ser Leu Glu Ser

1220 1225 1230

Leu Ser Met Ala Ile Arg Phe Ala Leu Lys Glu Thr Ser His Tyr Thr

1235 1240 1245

Ser Ser Glu Gly Asn Met Met His Ile Ala Leu Leu Gly Ala Asp Asn

1250 1255 1260

Lys Met His Ile Ile Gln Glu Ser Gly Asp Asp Ala Asp Arg Ile Ala

1265 1270 1275 1280

Lys Leu Pro Leu Ile Leu Lys Asp Asn Val Thr Asp Leu His Ala Ser

1285 1290 1295

Gly Val Lys Thr Ile Ser Phe Ile Val Gln Arg Asp Glu Ala Arg Met

1300 1305 1310

Thr Met Arg Arg Thr Phe Leu Trp Ser Asp Glu Lys Leu Ser Tyr Glu

1315 1320 1325

Glu Glu Pro Ile Leu Arg His Val Glu Pro Pro Leu Ser Ala Leu Leu

1330 1335 1340

Glu Leu Asp Lys Leu Lys Val Lys Gly Tyr Asn Glu Met Lys Tyr Thr

1345 1350 1355 1360

Pro Ser Arg Asp Arg Gln Trp His Ile Tyr Thr Leu Arg Asn Thr Glu

1365 1370 1375

Asn Pro Lys Met Leu His Arg Val Phe Phe Arg Thr Leu Val Arg Gln

1380 1385 1390

Pro Ser Val Ser Asn Lys Phe Ser Ser Gly Gln Ile Gly Asp Met Glu

1395 1400 1405

Val Gly Ser Ala Glu Glu Pro Leu Ser Phe Thr Ser Thr Ser Ile Leu

1410 1415 1420

Arg Ser Leu Met Thr Ala Ile Glu Glu Leu Glu Leu His Ala Ile Arg

1425 1430 1435 1440

Thr Gly His Ser His Met Tyr Leu His Val Leu Lys Glu Gln Lys Leu

1445 1450 1455

Leu Asp Leu Val Pro Val Ser Gly Asn Thr Val Leu Asp Val Gly Gln

1460 1465 1470

Asp Glu Ala Thr Ala Tyr Ser Leu Leu Lys Glu Met Ala Met Lys Ile

1475 1480 1485

His Glu Leu Val Gly Ala Arg Met His His Leu Ser Val Cys Gln Trp

1490 1495 1500

Glu Val Lys Leu Lys Leu Asp Cys Asp Gly Pro Ala Ser Gly Thr Trp

1505 1510 1515 1520

Arg Ile Val Thr Thr Asn Val Thr Ser His Thr Cys Thr Val Asp Ile

1525 1530 1535

Tyr Arg Glu Met Glu Asp Lys Glu Ser Arg Lys Leu Val Tyr His Pro

1540 1545 1550

Ala Thr Pro Ala Ala Gly Pro Leu His Gly Val Ala Leu Asn Asn Pro

1555 1560 1565

Tyr Gln Pro Leu Ser Val Ile Asp Leu Lys Arg Cys Ser Ala Arg Asn

1570 1575 1580

Asn Arg Thr Thr Tyr Cys Tyr Asp Phe Pro Leu Ala Phe Glu Thr Ala

1585 1590 1595 1600

Val Arg Lys Ser Trp Ser Ser Ser Thr Ser Gly Ala Ser Lys Gly Val

1605 1610 1615

Glu Asn Ala Gln Cys Tyr Val Lys Ala Thr Glu Leu Val Phe Ala Asp

1620 1625 1630

Lys His Gly Ser Trp Gly Thr Pro Leu Val Gln Met Asp Arg Pro Ala

1635 1640 1645

Gly Leu Asn Asp Ile Gly Met Val Ala Trp Thr Leu Lys Met Ser Thr

1650 1655 1660

Pro Glu Phe Pro Ser Gly Arg Glu Ile Ile Val Val Ala Asn Asp Ile

1665 1670 1675 1680

Thr Phe Arg Ala Gly Ser Phe Gly Pro Arg Glu Asp Ala Phe Phe Glu

1685 1690 1695

Ala Val Thr Asn Leu Ala Cys Glu Lys Lys Leu Pro Leu Ile Tyr Leu

1700 1705 1710

Ala Ala Asn Ser Gly Ala Arg Ile Gly Ile Ala Asp Glu Val Lys Ser

1715 1720 1725

Cys Phe Arg Val Gly Trp Ser Asp Asp Gly Ser Pro Glu Arg Gly Phe

1730 1735 1740

Gln Tyr Ile Tyr Leu Ser Glu Glu Asp Tyr Ala Arg Ile Gly Thr Ser

1745 1750 1755 1760

Val Ile Ala His Lys Met Gln Leu Asp Ser Gly Glu Ile Arg Trp Val

1765 1770 1775

Ile Asp Ser Val Val Gly Lys Glu Asp Gly Leu Gly Val Glu Asn Ile

1780 1785 1790

His Gly Ser Ala Ala Ile Ala Ser Ala Tyr Ser Arg Ala Tyr Lys Glu

1795 1800 1805

Thr Phe Thr Leu Thr Phe Val Thr Gly Arg Thr Val Gly Ile Gly Ala

1810 1815 1820

Tyr Leu Ala Arg Leu Gly Ile Arg Cys Ile Gln Arg Leu Asp Gln Pro

1825 1830 1835 1840

Ile Ile Leu Thr Gly Tyr Ser Ala Leu Asn Lys Leu Leu Gly Arg Glu

1845 1850 1855

Val Tyr Ser Ser His Met Gln Leu Gly Gly Pro Lys Ile Met Ala Thr

1860 1865 1870

Asn Gly Val Val His Leu Thr Val Ser Asp Asp Leu Glu Gly Val Ser

1875 1880 1885

Asn Ile Leu Arg Trp Leu Ser Tyr Val Pro Ala Tyr Ile Gly Gly Pro

1890 1895 1900

Leu Pro Val Thr Thr Pro Leu Asp Pro Pro Asp Arg Pro Val Ala Tyr

1905 1910 1915 1920

Ile Pro Glu Asn Ser Cys Asp Pro Arg Ala Ala Ile Arg Gly Val Asp

1925 1930 1935

Asp Ser Gln Gly Lys Trp Leu Gly Gly Met Phe Asp Lys Asp Ser Phe

1940 1945 1950

Val Glu Thr Phe Glu Gly Trp Ala Lys Thr Val Val Thr Gly Arg Ala

1955 1960 1965

Lys Leu Gly Gly Ile Pro Val Gly Val Ile Ala Val Glu Thr Gln Thr

1970 1975 1980

Met Met Gln Thr Ile Pro Ala Asp Pro Gly Gln Leu Asp Ser Arg Glu

1985 1990 1995 2000

Gln Ser Val Pro Arg Ala Gly Gln Val Trp Phe Pro Asp Ser Ala Thr

2005 2010 2015

Lys Thr Ala Gln Ala Leu Leu Asp Phe Asn Arg Glu Gly Leu Pro Leu

2020 2025 2030

Phe Ile Leu Ala Asn Trp Arg Gly Phe Ser Gly Gly Gln Arg Asp Leu

2035 2040 2045

Phe Glu Gly Ile Leu Gln Ala Gly Ser Thr Ile Val Glu Asn Leu Arg

2050 2055 2060

Thr Tyr Asn Gln Pro Ala Phe Val Tyr Ile Pro Met Ala Ala Glu Leu

2065 2070 2075 2080

Arg Gly Gly Ala Trp Val Val Val Asp Ser Lys Ile Asn Pro Asp Arg

2085 2090 2095

Ile Glu Cys Tyr Ala Glu Arg Thr Ala Lys Gly Asn Val Leu Glu Pro

2100 2105 2110

Gln Gly Leu Ile Glu Ile Lys Phe Arg Ser Glu Glu Leu Gln Asp Cys

2115 2120 2125

Met Ser Arg Leu Asp Pro Thr Leu Ile Asp Leu Lys Ala Lys Leu Glu

2130 2135 2140

Val Ala Asn Lys Asn Gly Ser Ala Asp Thr Lys Ser Leu Gln Glu Asn

2145 2150 2155 2160

Ile Glu Ala Arg Thr Lys Gln Leu Met Pro Leu Tyr Thr Gln Ile Ala

2165 2170 2175

Ile Arg Phe Ala Glu Leu His Asp Thr Ser Leu Arg Met Ala Ala Lys

2180 2185 2190

Gly Val Ile Lys Lys Val Val Asp Trp Glu Glu Ser Arg Ser Phe Phe

2195 2200 2205

Tyr Lys Arg Leu Arg Arg Arg Ile Ser Glu Asp Val Leu Ala Lys Glu

2210 2215 2220

Ile Arg Ala Val Ala Gly Glu Gln Phe Ser His Gln Pro Ala Ile Glu

2225 2230 2235 2240

Leu Ile Lys Lys Trp Tyr Ser Ala Ser His Ala Ala Glu Trp Asp Asp

2245 2250 2255

Asp Asp Ala Phe Val Ala Trp Met Asp Asn Pro Glu Asn Tyr Lys Asp

2260 2265 2270

Tyr Ile Gln Tyr Leu Lys Ala Gln Arg Val Ser Gln Ser Leu Ser Ser

2275 2280 2285

Leu Ser Asp Ser Ser Ser Asp Leu Gln Ala Leu Pro Gln Gly Leu Ser

2290 2295 2300

Met Leu Leu Asp Lys Met Asp Pro Ser Arg Arg Ala Gln Leu Val Glu

2305 2310 2315 2320

Glu Ile Arg Lys Val Leu Gly

2325

<210> 5

<211> 25

<212> DNA

<213> ACCase wild type Sequence ACC-CF primer (Artificial Sequence)

<400> 5

ATGACATCCACACATGTGGCGACAT 25

<210> 6

<211> 25

<212> DNA

<213> ACCase wild type Sequence ACC-CR primer (Artificial Sequence)

<400> 6

TCAACCAAGGACCTTCCTGATTTCT 25

Claims (8)

1. A rice ACCase mutant protein is characterized in that the amino acid sequence of the protein corresponds to that of wild rice ACCase, arginine is mutated into cysteine from position 1731 and/or tyrosine is mutated into aspartic acid from position 2276, and the amino acid sequence is shown as SEQ ID NO.2 or SEQ ID NO:4 or SEQ ID NO: and 6.

2. An ACCase mutant gene encoding the protein of claim 1.

3. The ACCase mutant gene according to claim 2, wherein the nucleotide sequence is selected from SEQ ID No.1 or SEQ ID NO:3 or SEQ ID NO: 5.

4. the ACCase mutant gene according to claim 2 or 3, wherein the introduction of the gene into wild-type rice confers resistance to a herbicide to a recipient rice, the herbicide being galileo.

5. The ACCase mutant gene according to claim 4, wherein the gene is introduced into recipient rice by transgenic, crossing or backcrossing.

6. The ACCase mutant gene according to claim 5, wherein said transgenic method comprises the steps of:

1) constructing an expression vector containing an ACCase mutant gene;

2) genetically transforming rice with the expression vector to make rice contain the ACCase mutant gene according to claim 2 or 3;

3) expressing rice ACCase mutant protein according to claim 1.

7. A recombinant vector comprising the ACCase mutant gene according to claim 2 or 3.

8. A host cell comprising an expression vector of an ACCase mutant gene according to claim 2 or 3.

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