EcCYP89B16 gene, polypeptide, carrier and application of EcCYP89B16 gene in plant herbicide resistance and herbicide-resistant transgenic plant breeding

Abstract

The invention discloses an EcCYP89B16 gene, a polypeptide, a carrier and an application of the EcCYP89B16 gene in plant herbicide resistance and herbicide-resistant transgenic plant breeding. The nucleotide sequence of the gene is SEQ ID NO: 1 or a sequence with not less than 80% of identity compared with the SEQ ID NO: 1. The invention also discloses a protein polypeptide coded by the gene. The amino acid sequence of the protein polypeptide is SEQ ID NO: 2 or a sequence with not less than 80% of identity compared with the SEQ ID NO: 2. The invention relates to the herbicide-resistant gene, a protein coded by the herbicide-resistant gene, a carrier and application of the herbicide-resistant gene and the carrier in the aspect of herbicide-resistant transgenic plant breeding. The P450 gene can be expressed in plants to enable the plants to obtain resistance to aryloxyphenoxypropionic acid herbicides, so that weeds can be selectively prevented and removed by using the herbicides.

Description

Technical Field

[0001] The present invention relates to the technical field of plant genetic engineering, and in particular to a herbicide-resistant gene and its encoded protein and vector, as well as the application of the gene and vector in the breeding of herbicide-resistant transgenic plants. Background Art

[0002] With the widespread adoption of direct seeding in agricultural production, weeds in paddy fields have become a significant factor affecting crop yields. In particular, the physiological and metabolic processes of grass weeds in paddy fields are highly similar to those of rice, leading to a lack of appropriate herbicides for control. Because traditional breeding and domestication processes do not address herbicide tolerance, resistance is extremely rare in natural genetic resources. Using herbicide-inhibited target genes to screen and identify endogenous resistance mutants is currently a viable approach for developing herbicide-resistant germplasm in staple crops. For example, EMS-induced mutagenesis of the imidazole herbicide target gene ALS has yielded resistant resources. In recent years, this mutation has been widely used in varieties such as "Jinjing 818," demonstrating significant advantages in chemical weed control. However, endogenous resistance loci in key herbicide target genes, such as EPSPS, ACCase, ALS, and HPPD, are often patented by foreign commercial companies. Furthermore, with repeated application of a single herbicide, acquired resistance in field weeds is rapidly increasing. Therefore, it is very necessary to discover new herbicide resistance genes in plants and express heterologous metabolic genes through transgenic means to obtain herbicide resistance traits for the cultivation of herbicide-resistant plants.

[0003] Aryloxyphenoxy propionate (APP) herbicides are a key class of herbicides for controlling grass weeds. They were developed in the 1960s by Hoechst, who replaced the phenyl group in the 2,4-D structure with diphenyl ether and further developed the compound diclofop-butyl. In 1971, the structural model for this class of herbicides was established, with the parent ring consisting of A and B. Since 1975, many pesticide companies have modified and remodeled this original model, replacing the A benzene ring with a heterocyclic or fused ring and introducing active groups such as F atoms into the rings, resulting in a series of herbicides with increased activity and selectivity.

[0004] APP herbicides primarily inhibit the activity of acetyl-CoA carboxylase (ACCase, EC6.4.1.2) in grasses, thereby inhibiting the synthesis of fatty acids. Fatty acids are the primary energy storage and supply substances in plants, key components of cell membranes, and play an important role in plant catalytic metabolism and biosynthesis.

[0005] Take cyhalofop-butyl, for example. It is the only APP herbicide with a high safety profile for rice. Like other APP herbicides, it is a systemic herbicide. It is absorbed by the plant's leaves and sheaths, transported through the phloem, and accumulated in the plant's meristematic zone. It inhibits the enzyme acetyl-CoA carboxylase (ACCase), halting fatty acid synthesis, disrupting normal cell growth and division, and damaging lipid-containing structures such as membranes, ultimately leading to plant death. Cyhalofop-butyl is primarily used to control grass weeds such as barnyardgrass, chinensis, crabgrass, foxtail grass, goosegrass, and paspalum distachyon in rice fields. The process from weed absorption to weed death is relatively slow, generally taking 1-3 weeks. Symptoms of weed damage appear 5-7 days after application, starting with yellowing and browning in the center of the leaves, then gradually spreading throughout the plant, ultimately leading to plant death. However, cyhalofop-butyl is toxic to crops other than rice, and its long-term, widespread use has also led to a degree of environmental toxicity, significantly limiting its scope and duration of use. Therefore, identifying herbicide-resistant genes and conferring herbicide resistance in crops through genetic engineering are of great significance.

[0006] Cytochrome P450 (CYP) enzymes are a class of membrane-bound hemoglobin enzymes widely found in microorganisms, plants, animals, and humans. They possess diverse biocatalytic activities, including oxidation, epoxidation, hydroxylation, and demethylation. P450s are particularly important in drug metabolism, participating in the metabolism of drugs and toxins in humans, the biosynthesis of secondary metabolites in plants and microorganisms, and several microbial degradation pathways. This has attracted considerable attention from pesticide researchers. Currently, several CYP81 subfamily genes (CYP81A12, CYP81A21, CYP81A68, CYP81A10v7, and CYP81Q32) have been shown to metabolize herbicides, conferring herbicide resistance in weeds. Of these, only CYP81A68 has been shown to confer weed resistance to cyhalofop-butyl. However, non-CYP81 family genes that metabolize cyhalofop-butyl and confer cyhalofop-butyl resistance in weeds have not been identified. Summary of the Invention

[0007] The primary purpose of the present invention is to provide a herbicide-resistant gene that can be efficiently expressed in plants. By transferring the gene sequence into plants, new herbicide-tolerant transgenic plants can be cultivated.

[0008] The present invention provides a herbicide-resistant gene EcCYP89B16, the nucleotide sequence of which is SEQ ID NO: 1 or a sequence having an identity of not less than 80% with SEQ ID NO: 1.

[0009] The second aspect of the present invention is to provide a protein polypeptide encoded by the gene SEQ ID NO: 1, wherein the amino acid sequence of the protein polypeptide is SEQ ID NO: 2 or a sequence having an identity of not less than 80% with SEQ ID NO: 2.

[0010] The third aspect of the present invention aims to provide the use of the gene EcCYP89B16 in plant resistance to herbicides.

[0011] Specifically, the EcCYP89B16 gene is expressed in plants, thereby enabling the plants to acquire herbicide resistance.

[0012] The herbicide is an aryloxyphenoxypropionic acid herbicide, specifically including at least one of cyhalofop-butyl, pinoxaden, and penoxsulam.

[0013] The plants include rice, wheat, corn, cotton, sorghum or forage grass.

[0014] The fourth aspect of the present invention also provides a plasmid comprising an expression cassette formed by connecting the nucleotide sequence of the gene and a nucleotide sequence for controlling expression, wherein the nucleotide sequence encodes the above-mentioned protein polypeptide.

[0015] The fifth aspect of the present invention aims to provide the use of the gene, the polypeptide, or the plasmid in the breeding of herbicide-resistant transgenic plants.

[0016] The nucleotide sequence of the herbicide-resistant gene of the present invention can have a variety of variations, including but not limited to: 1) different nucleotide sequences obtained by utilizing different codons for the same amino acid, these sequences encoding proteins with the same activity; 2) nucleotide sequences that introduce nucleotide sequence variations while still encoding a protein with herbicide resistance. Such variations can be random, targeted, or insertional or deletional. Those skilled in the art can generate these variations using molecular biology methods.

[0017] Using the nucleotide sequences provided by the present invention, homologous genes with the same function can also be obtained. One method is to use the nucleic acids provided by the present invention as probes to hybridize a DNA library to obtain homologous genes. Another method is to clone homologous genes by PCR using primers designed based on the nucleic acid sequences provided by the present invention. Furthermore, those skilled in the art can also use molecular informatics methods to identify genes with high homology from genomic libraries using the nucleic acid sequences and protein sequences provided by the present invention. For example, using the BLAST method (www.ncbi.nih.gov) based on the nucleotide sequences provided by the present invention and the amino acid sequences of protein polypeptides provided by the present invention, genes with relatively high homology to the genes provided by the present invention can be discovered.

[0018] The herbicide-resistant gene nucleotide sequences provided by the present invention can be used to construct artificial genes capable of being expressed in plants. Similarly, the cyhalofop-butyl protein polypeptide sequences provided by the present invention can be used to synthesize nucleic acid sequences and further construct artificial genes capable of being expressed in plants. The components of artificial genes capable of being expressed in plants include a promoter, a herbicide-resistant gene, and a terminator.

[0019] The promoters and terminators required for expressing and conferring herbicide resistance in plants are already available. For example, when transforming monocots, the promoter can be the maize Ubiqutin-1 promoter or the rice Actin promoter; the terminator can be the Agrobacterium tumefaciens terminator (Nos) or other terminators. This expression construct can be integrated into the plant genome using Agrobacterium (such as an Agrobacterium strain), a gene gun, or other methods to produce cyhalofop-butyl-resistant transgenic plants. Plant transformation techniques and methods are known and established. Transformation methods and procedures vary for different plants. However, typically, the construct is introduced into immature embryos, mature embryos, undifferentiated callus, or protoplasts of the plant using Agrobacterium or a gene gun. Transformed shoots are then differentiated to obtain transformed shoots, which are then cultured in rooting medium to obtain plantable transgenic seedlings. Herbicide-resistant transgenic plants can be screened for herbicide resistance by spraying. Furthermore, cyhalofop-butyl-resistant transgenic plants can be screened by spraying.

[0020] The present invention is applicable to all plants, including dicotyledonous and monocotyledonous plants.

[0021] The present invention first discovered a cyhalofop-butyl-resistant barnyardgrass variety, then used its genomic sequence to clone its cytochrome P450 gene, EcCYP89B16. Furthermore, the EcCYP89B16 gene's ability to resist cyhalofop-butyl and other aryloxyphenoxypropionic acid herbicides was demonstrated in rice, along with its potential use in cultivating transgenic aryloxyphenoxypropionic acid-resistant crops. The present invention relates to a novel cytochrome P450 protein gene, EcCYP89B16, whose resistance to aryloxyphenoxypropionic acid herbicides has never been reported. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] The specific embodiments of the present invention are further described in detail below with reference to the accompanying drawings.

[0023] Figure 1This is a schematic diagram of the construction of a plant expression vector; the expression cassette is mainly driven by the Ubi ubiquitin promoter to overexpress EcCYP89B16. In addition, there is a 35S promoter-driven HPT (hygromycin phosphotransferase), which can confer hygromycin resistance to rice callus and is used for screening positive calli. Kan confers resistance to kanamycin on the vector and is used for positive screening during vector construction.

[0024] Figure 2 The results of RT-qPCR identification of differentially expressed P450 genes screened from the sensitive barnyardgrass population in Example 2 are as follows;

[0025] Figure 3 The results of the cyhalofop-butyl tolerance test of transgenic rice expressing the barnyardgrass gene EcCYP89B16 in Example 3 are as follows;

[0026] Figure 4 The results of the tolerance test of the transgenic rice with the barnyardgrass gene EcCYP89B16 to pinoxaden in Example 4 are as follows;

[0027] Figure 5 The results of the tolerance test of transgenic rice with the barnyardgrass gene EcCYP89B16 to penoxsulam in Example 5 are shown. DETAILED DESCRIPTION

[0028] The present invention will be further described below in conjunction with specific examples, but the scope of protection of the present invention is not limited thereto. It should be understood that these examples are merely intended to illustrate the method of the present invention and are not intended to limit the scope of the present invention. Any experimental method without specific experimental conditions or special instructions is a conventional condition and conventional method well known to those skilled in the art.

[0029] Example 1 Determination of resistance of cyhalofop-butyl-resistant barnyardgrass

[0030] Echinochloa crusgalli (Echinochloa crusgalli) belongs to the Poaceae family and is an annual weed. It is one of the 18 malignant weeds worldwide and one of the most widespread and harmful weeds in my country's rice fields. In order to clarify the resistance level of Echinochloa crusgalli to cyhalofop-butyl, the sensitivity of the collected Echinochloa crusgalli biotypes to cyhalofop-butyl was studied using the whole plant assay method. The results showed that the fresh weight inhibitory dose (EDD) of cyhalofop-butyl to resistant (R) Echinochloa crusgalli biotypes was 1.34 mmol / l. 50 ) value is 167.54ga.i.ha -1 ; while ED for sensitive (S) Echinochloa crusgalli biotype 50 The value is 55.16g aiha -1 The relative resistance multiple was 3.05-fold. Therefore, the resistant (R) barnyardgrass biotype may contain cyhalofop-butyl resistance genes.

[0031] Example 2 Cloning of resistance genes

[0032] A plant's resistance to cyhalofop-butyl may be due to various mechanisms, such as the presence of a cyhalofop-butyl-resistant ACCase gene; the presence of an enzyme capable of degrading cyhalofop-butyl; the ability to restrict cyhalofop-butyl's uptake; or the presence of an enzyme that modifies cyhalofop-butyl, rendering it inactive. To investigate the mechanisms of target resistance in resistant barnyardgrass populations, 20 strains from each resistant population were cloned for the acetyl-CoA carboxylase (ACCase, EC 6.4.1.2) gene fragment. The resulting gene sequence was then compared with that of a susceptible (S) barnyardgrass population. The R population was found to have no target enzyme mutations, indicating that this population lacks a target enzyme resistance mechanism. Transcriptome sequencing successfully generated transcriptome data for two barnyardgrass populations (R and S). Using the barnyardgrass genome database as a reference genome, statistical evaluation of sequencing quality revealed that both had base Q30 values ​​exceeding 90%, indicating high sequencing quality. Further analysis of the assembled gene expression levels revealed 351 differentially expressed P450 genes between R and S, and screened out nine potential non-target resistance-related genes. Fluorescence quantitative PCR validation of these nine non-target resistance-related differentially expressed genes using RNA samples from transcriptome sequencing revealed that the cytochrome P450 gene (EcCYP89B16) was consistently highly expressed in the resistant population R compared to the sensitive population S. Using the extracted total RNA of Echinochloa crusgalli as a template, based on the EST sequence fragment of the EcCYP89B16 gene of Echinochloa crusgalli obtained from the transcriptome, two specific primers were designed according to the 3' and 5' end splicing sequences for PCR amplification and cloning to obtain the EcCYP89B16 plasmid (the empty vector was pClone007 Versatile Simple Vector K, Beijing Qingke) (primer sequences were (F: 5'-ATGGACACGCAGCTCCTACTCC-3', R: 5'-TCACCCAGAGCCAGAGGTTCT-3') and sequencing, and finally the full-length EcCYP89B16 gene of Echinochloa crusgalli was successfully obtained (SEQ ID NO: 1).

[0033] Example 3 Construction of EcCYP89B16 gene overexpression vector and acquisition of transgenic cyhalofop-butyl-resistant rice

[0034] 1) Construct EcCYP89B16 gene overexpression vector.

[0035] Specific primers (pox-F: 5'-gtgttacttctgcagggtaccATGGACACGCAGCTCCTACTCC-3', pox-R: 5'-gttatcggatccataacgcgtTCACCCAGAGCCAGAGGTTCT-3') were designed, and the EcCYP89B16 plasmid cloned and sequenced correctly in Example 2 was used as a template. High-fidelity enzyme ( Max DNA Polymerase (purchased from TaKaRa) was used for PCR amplification and purification to obtain the EcCYP89B16 fragment with pox homology arms.

[0036] The pox vector was double-digested using Thermo Fisher FastDigest (FastDigest KpnI: 5'GGTAC↓C 3' and FastDigestMluI: 5'A↓CGCGT 3'). The specific implementation steps were as follows: 2 μL 10XFastDigest Green Buffer, 1 μL FastDigest KpnI, 1 μL FastDigest MluI, 1000 ng pox plasmid were added to a PCR tube, and sterile water was added to 20 μL. After reaction at 37°C for 5 minutes, the linearized vector was purified.

[0037] The purified target fragment and linearized vector were recombined and ligated using the ClonExpressII One Step Cloning Kit. 10 μL of the recombinant product was transformed into DH5α competent Escherichia coli, and a single clone was picked for sequencing identification to obtain the expression vector pox-EcCYP89B16 containing the full-length expression sequence of the EcCYP89B16 gene of barnyardgrass.

[0038] 2) Transformation of Agrobacterium with overexpression vector

[0039] Agrobacterium EHA105 was used to transform the overexpression vector into rice callus for overexpression to test whether EcCYP89B16 has herbicide resistance. The overexpression vector was transformed into Agrobacterium as follows:

[0040] Soak the electric shock cup and cup lid in 75% alcohol for 2 hours for disinfection. Take it out of the clean bench and place it on clean filter paper to drain the water. After the alcohol is completely evaporated, insert it into ice for pre-cooling.

[0041] Take EHA105 Agrobacterium competent cells (Beijing Qingke) from -80℃ refrigerator, pinch them gently with your fingers until they partially melt, then place them on ice to completely melt;

[0042] Pipette 1000 ng of recombinant plasmid pox-EcCYP89B16 into the EHA105 competent medium, pipette gently to mix, and then transfer the mixture completely into the electroporation cup and cover the cup.

[0043] The parameters of the electroporator (Eppendorf, Germany) were set to 2.5 kV and 5 ms. The water on the electroporation cup was wiped dry and quickly placed in the electroporation tank. After the electroporation, it was quickly placed on ice. 700 μL of antibiotic-free LB medium was added, pipetted and mixed, and then the whole was transferred to a 1.5 mL tube and incubated on a shaker at 28°C for 2-3 hours.

[0044] Centrifuge at 4000 rpm for 1 minute to collect the cells, retain about 100 μL of supernatant, mix with the cells by pipetting, and then spread on LB medium containing 50 μg / mL kanamycin and 20 μg / mL rifampicin. Incubate inverted at 28°C for 2-3 days.

[0045] Pick a single colony for colony PCR to determine whether the plasmid has been successfully transferred and select positive transformants for future use.

[0046] 3) Induction of rice callus

[0047] ZH11 rice seeds were shelled and mature and healthy seeds were selected for disinfection and induction.

[0048] The specific steps are as follows:

[0049] Place rice seeds in a 50 mL centrifuge tube, add an appropriate amount of 75% ethanol, shake for about 60 seconds, pour out the ethanol, and wash with sterile water 5-7 times;

[0050] Add 2.5% sodium hypochlorite and 1 drop of Tween 20 to a 50 mL centrifuge tube, shake for 15 minutes, pour out the sodium hypochlorite and wash with sterile water 5-7 times;

[0051] Add an appropriate amount of 2.5% sodium hypochlorite again, shake for 15 minutes, pour out the sodium hypochlorite and wash with sterile water 7-10 times;

[0052] The seeds were taken out and placed on filter paper to absorb moisture, and then placed on N6D induction medium and induced at 30°C in the dark for 10-12 days.

[0053] 4) Agrobacterium infection of callus and co-cultivation.

[0054] The positive EHA105 transformants from step 2) were shaken in LB liquid medium until turbid. The transformants were then added to 50 mL of LB medium at a ratio of 1:50 and vigorously shaken until the OD600 was approximately 0.6. The cells were collected and diluted to OD600 = 0.1 using AAM+AS medium. The dense and firm calli from step 3) were selected and added to the medium. After soaking for 2 minutes, the cells were removed and placed on filter paper to absorb the moisture.

[0055] A piece of filter paper was placed on the 2N6 medium, 500 μL of AAM+AS was added dropwise, and the above-mentioned dried callus was placed on the filter paper and cultured in the dark at 30°C for 3 days.

[0056] 5) Positive callus screening and cyhalofop-butyl sensitivity determination

[0057] The co-cultured callus tissue was transferred to a 50 mL tube, washed 10 times with sterile water, and then washed 7 times with sterile water containing cefotaxime (250 mg / L) and hygromycin (50 mg / L). After drying on filter paper, the callus was transferred to N6D medium containing hygromycin (50 mg / L), cefotaxime (250 mg / L), and timentin (200 mg / L) to select positive calli. The calli were cultured at 30°C in the dark and transferred to new N6D medium for subculture after 14 days.

[0058] After two rounds of screening, newly grown dense, hard, milky yellow positive calli were selected and transferred to a differentiation medium containing hygromycin (50 mg / L). They were first cultured in the dark for 3 days and then transferred to light conditions (30°C, 1500-2000xl of light, photoperiod: 16h / 8h (light / dark)). After about 15-25 days, green spots appeared, and seedlings were further differentiated after 30-40 days.

[0059] Screening of transgenic plants. When buds differentiated from resistant callus reach approximately 2 cm in length, transfer the seedlings to rooting medium and culture for approximately two weeks. Select seedlings approximately 10 cm tall with well-developed root systems, wash off the medium, and transplant into soil in a greenhouse to obtain transformed plants.

[0060] Transgenic rice was treated with cyhalofop-butyl and the transgenic rice with pox-GFP overexpression was used as a control to observe its sensitivity to cyhalofop-butyl. The results showed that the transgenic rice with pox-GFP overexpression (used as a sensitive strain as a control to confirm that the resistance of rice was enhanced after EcCYP89B16 overexpression) was sensitive to cyhalofop-butyl at a cyhalofop-butyl concentration of 4200 g aiha -1 The transgenic rice with overexpression of pox-EcCYP89B16 gene of Echinochloa crus-galli was completely dead at 4200 g aiha. -1 Therefore, we concluded that overexpression of the EcCYP89B16 gene in barnyardgrass could make transgenic rice resistant to cyhalofop-butyl.

[0061] Example 4 Determination of Sensitivity of EcCYP89B16 Transgenic Rice to Pinoxaden

[0062] The transgenic rice was treated with pinoxaden, and the transgenic rice with pox-GFP overexpression was used as a control to observe its sensitivity to pinoxaden. The results showed that the transgenic rice with pox-GFP overexpression was sensitive to pinoxaden at a concentration of 24 g a.i.ha -1 The transgenic rice with overexpression of pox-EcCYP89B16 gene of Echinochloa crus-galli was completely dead at 24 ga.i.ha. -1 Therefore, we concluded that overexpression of the EcCYP89B16 gene in barnyardgrass could make transgenic rice resistant to pinoxaden.

[0063] Example 5 Determination of Sensitivity of EcCYP89B16 Transgenic Rice to Penoxsulam

[0064] Transgenic rice was treated with penoxsulam and the transgenic rice with pox-GFP overexpression was used as a control to observe its sensitivity to penoxsulam. The results showed that the transgenic rice with pox-GFP overexpression was sensitive to penoxsulam at a penoxsulam concentration of 300g aiha -1 was severely inhibited, while the transgenic rice with overexpression of pox-EcCYP89B16 gene of Echinochloa crus-galli was -1 There was only slight inhibition at the concentration of penoxsulam, so we concluded that overexpression of the EcCYP89B16 gene in barnyardgrass can make transgenic rice resistant to penoxsulam.

[0065] Finally, it should be noted that the above examples are merely specific embodiments of the present invention. Obviously, the present invention is not limited to the above examples and is susceptible to numerous variations. It should be noted that those skilled in the art would be able to make numerous improvements and modifications without departing from the principles of the present invention, and such improvements and modifications should be considered within the scope of protection of the present invention.

[0066] The present invention uses culture medium and formula

[0067] N6D medium protocol (callus induction and growth medium):

[0068]

[0069]

[0070]

[0071] AAM medium

[0072]

[0073] 2N6-AS medium (without AS)

[0074]

[0075] Regeneration medium (differentiation medium)

[0076]

[0077]

[0078] MS hormon free medium (rooting medium)

[0079]

[0080] Antibiotic working solution concentration:

[0081]

[0082] Hormone working solution concentration:

[0083]

[0084]

[0085]

Claims

1. A herbicide resistance gene EcCYP89B16, characterized in that: The nucleotide sequence is SEQ ID NO: 1 or a sequence having no less than 80% identity with SEQ ID NO:

1.

2. The polypeptide encoded by the herbicide-resistant gene EcCYP89B16 according to claim 1, characterized in that: The amino acid sequence is SEQ ID NO: 2 or a sequence having no less than 80% identity with SEQ ID NO:

2.

3. Use of the gene EcCYP89B16 according to claim 1 in plant resistance to herbicides.

4. The use according to claim 3, characterized in that: By expressing the EcCYP89B16 gene in plants, the plants can acquire the ability to resist herbicides.

5. The use according to claim 3, characterized in that: The herbicide is an aryloxyphenoxypropionic acid herbicide, specifically including at least one of cyhalofop-butyl, pinoxaden, and penoxsulam.

6. The use according to claim 3, characterized in that: The plants include rice, wheat, corn, cotton, sorghum or forage grass.

7. A plasmid, characterized in that: An expression frame comprising the nucleotide sequence of claim 1 connected to a nucleotide sequence for controlling expression, wherein the nucleotide sequence encodes the protein polypeptide of claim 2.

8. Use of the plasmid according to claim 7 in plant resistance to herbicides, characterized in that: The plasmid is transferred into plants for expression, thereby enabling the plants to acquire the ability to resist herbicides.

9. The use according to claim 8, characterized in that: The herbicide is an aryloxyphenoxypropionic acid herbicide, specifically including at least one of cyhalofop-butyl, pinoxaden, and penoxsulam; the plants include rice, wheat, corn, cotton, sorghum or forage.

10. Use of the gene according to claim 1, the polypeptide according to claim 2, or the plasmid according to claim 7 in breeding herbicide-resistant transgenic plants.

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