LmSCPL18 gene, peptides, vectors, and their application in enhancing herbicide resistance in plants and breeding herbicide-resistant transgenic plants.

Abstract

This invention belongs to the field of gene biology technology, specifically involving LmSCPL18 Genes, peptides, vectors, and their applications in enhancing herbicide resistance in plants and in the breeding of herbicide-resistant transgenic plants. A LmSCPL18 Genes, the ones mentioned LmSCPL18 The nucleotide sequence of the gene is shown in SEQ ID NO.1 of the sequence listing. This invention utilizes cloned *Lycium chinense*. LmSCPL18 Genes were transferred into rice, and the resistance levels of the transgenic rice to different herbicides were measured. The results showed that the transgenic ryegrass... LmSCPL18 Genetically modified rice exhibits significant resistance to the ACCase inhibitor herbicide clodinafop-propargyl and the ALS inhibitor herbicide sulfadiazine. Therefore, ryegrass... LmSCPL18 Genes can enhance plant resistance to herbicides and lay the foundation for breeding herbicide-resistant transgenic plants, which is of great significance.

Description

Technical Field

[0001] This invention belongs to the field of gene biology technology, specifically involving LmSCPL18 Genes, peptides, vectors and their applications in improving plant herbicide resistance and in the breeding of herbicide-resistant transgenic plants. Background Technology

[0002] Weeds compete with crops for sunlight, water, and nutrients, affecting the safe production of crops. Herbicides are the most economical, effective, and convenient means of weed control in modern agricultural production, occupying an important position. Currently, herbicides include various types such as ACCase inhibitors and ALS inhibitors. ACCase inhibitors specifically inhibit the activity of acetyl-CoA carboxylase in plants, blocking the synthesis pathway of fatty acids and ultimately causing weed death. ALS inhibitors specifically inhibit the activity of acetolactate synthase in plants, blocking the synthesis pathway of branched-chain amino acids, thus achieving weed control.

[0003] However, the unscientific use of herbicides can cause phytotoxicity to crops, and different varieties of the same crop have varying tolerances to herbicides. Furthermore, the continuous and unscientific use of herbicides leads to the gradual accumulation and development of herbicide resistance in weeds, which in turn increases the dosage required, exacerbating the phytotoxic effects on crops and impacting crop yield and quality. Therefore, improving crop herbicide resistance and breeding herbicide-resistant genetically modified crops are of great significance.

[0004] The inventor discovered through experiments LmSCPL18 Overexpression of genes can lead to resistance in transgenic plants to ACCase inhibitor herbicides and ALS inhibitor herbicides. No reports on this have been found to date. Summary of the Invention

[0005] The purpose of this invention is to provide LmSCPL18 Genes, peptides, vectors and their applications in improving plant herbicide resistance and in the breeding of herbicide-resistant transgenic plants.

[0006] To achieve the above objectives, the present invention provides the following technical solution: A sort of LmSCPL18 Genes, the ones mentioned LmSCPL18 The nucleotide sequence of the gene is shown in SEQ ID NO.1 of the sequence listing, specifically:

[0007] The present invention also provides a protein polypeptide, wherein the protein polypeptide is composed of the... LmSCPL18 It is encoded by a gene, and its amino acid sequence is shown in SEQ ID NO.2 of the sequence listing, specifically: MDMPKQLLLLLVVVVTAASYGCPAAAAAARERNTITHVKGFDGPLPFSLETGYVEVDETHGAELFYYFIQSERSPQEDPLILWITGGPGCSALSGLLFEIGPLKFDVAGYTEGFPRL VYFEDSWTQVSNVIFLDAPVGTGFSYAREEQGLNVSLTGTGRQLRIFLQKWLAKHPEFASNPLYIGGDSYSGYTVPVTALEIANHPDGGLNLKGYLVGNAATDDKYDTGGKVPFMHG MGLISDELYEAAQGSCMGDFVSPPTNAQCANALQEISQATWAINPVHILEPMCGLAMRPTTTSNVLARRSARMLVQEQDMLRLPVECRDNGYRLSYIWADDAEVRETLGIREGSIG AWSRCTTLMHFRHDLRSTIPYHRNLTQRGYRGLVYNGDHDMDMTFVGTQAWIRTLGYPVVEPWRPWYANRQVAGFTTEYAYNLTFATVKGGGHTAPEYRPKECLAMLDRWTSTVGKI.

[0008] The present invention also provides a gene expression vector, the gene expression vector comprising the... LmSCPL18 Gene.

[0009] The present invention also provides the above. LmSCPL18 The application of the gene, the protein polypeptide, or the gene expression vector in improving the herbicide resistance of plants and in the breeding of herbicide-resistant transgenic plants.

[0010] Based on the above scheme, the herbicides include ACCase inhibitor herbicides and ALS inhibitor herbicides.

[0011] Based on the above scheme, the ACCase inhibitor herbicides include clodinafop-propargyl, quizalofop-p-ethyl, quizalofop-p-ethyl, quizalofop-p-ethyl, clethodim, clethodim, and clethodim.

[0012] Based on the above scheme, the ALS inhibitor herbicides include mesosulfuron-methyl, pyrazosulfuron-methyl, flusulfuron-methyl, methoxyfenozide, imidacloprid, and methyl methoxyfenozide.

[0013] Based on the above scheme, the plants include wheat, barley, oats, rice, forage grass, corn, millet, rye, perennial ryegrass, stiff ryegrass, and sorghum.

[0014] This invention also provides a method for improving the herbicide resistance of plants or for breeding herbicide-resistant transgenic plants, specifically: ... LmSCPL18 The nucleic acid sequence of a gene is constructed into a gene expression vector, transformed into plants, and expressed in the plants to improve the herbicide resistance of plants or to achieve the breeding of herbicide-resistant transgenic plants; wherein, the... LmSCPL18 The nucleotide sequence of the gene is shown in SEQ ID NO.1 of the sequence listing.

[0015] Based on the above scheme, the herbicides include ACCase inhibitor herbicides and ALS inhibitor herbicides; the plants include wheat, barley, oats, rice, forage grass, corn, millet, rye, perennial ryegrass, stiff ryegrass, and sorghum.

[0016] Compared with the prior art, the present invention has the following beneficial effects: This invention utilizes cloned ryegrass. LmSCPL18 Genes were transferred into rice, and the resistance levels of the transgenic rice to different herbicides were measured. The results showed that the transgenic ryegrass... LmSCPL18 Genetically modified rice exhibits significant resistance to the ACCase inhibitor herbicide clodinafop-propargyl and the ALS inhibitor herbicide sulfadiazine. Therefore, ryegrass... LmSCPL18 Genes can enhance plant resistance to herbicides and lay the foundation for breeding herbicide-resistant transgenic plants, which is of great significance. Attached Figure Description

[0017] Figure 1 for LmSCPL18 Transcriptome data validation; Figure 2 for LmSCPLl8 Gel electrophoresis image of a gene clone; Figure 3 for LmSCPLl8 Gel electrophoresis image of gene overexpression vector construction; Figure 4 In genetically modified rice LmSCPLl8 The relative expression level; Figure 5 The results show the sensitivity of genetically modified rice to herbicides.

[0018] Explanation of key figure labels: exist Figure 2 In the middle, a. LmSCPLl8a. Gene amplification, M: 2000 maker, 1: PCR amplification product; b. PMD-LmSCPLl8 colony PCR identification, M: 2000 maker, 1-9: PCR amplification products, -: negative control; Figure 3 In the diagram, a. pox empty vector double digestion, M: 5000 maker, 1: digested pox vector; b. colony PCR identification of pox-LmSCPLl8 after DH5α transformation, M: 2000 maker, 1-9: pox-LmSCPLl8 strain, -: negative control; c. colony PCR identification of pox-LmSCPLl8 after EHA105 transformation, M: 2000 maker, 1-6: PCR amplification products, -: negative control. Detailed Implementation

[0019] The technical solution of this invention patent will be clearly and completely described below. Obviously, the described embodiments are some embodiments of this invention, but not all embodiments.

[0020] 1. Materials and Methods 1.1 Sensitivity test of Lerium perfoliatum to herbicides Multiflora ryegrass ( Lolium multiflorum Italian ryegrass, also known as Italian ryegrass, is an annual or biennial grass. Italian ryegrass has a strong seed propagation capacity, a wide suitable growing range, and is easily spread. It can generally cause a 30% reduction in wheat yield, and in severely affected areas, it can cause complete crop failure, seriously jeopardizing the safe and stable production of wheat in my country. To clarify the sensitivity of Italian ryegrass to commonly used herbicides in wheat fields, the whole-plant bioassay method was used to study the sensitivity of different Italian ryegrass biotypes to commonly used ACCase inhibitor herbicides such as clodinafop-propargyl and clodinafop-propargyl, and ALS inhibitor herbicides such as mesosulfuron-methyl and disulfuron-methyl. See “Resistance of Italian ryegrass to ACCase and ALS inhibitors and analysis of target gene mutations in Henan Province” (Xu Hongle et al., Institute of Plant Protection, Henan Academy of Agricultural Sciences, Journal of Plant Protection, 2023, 50(1): 224-230).

[0021] The results showed that clodinafop-propargyl, ... mesosulfuron-methyl, and pyrazosulfuron-methyl resistant to the end-effector effect (ED) of resistant ryegrass. 50 The values ​​were 94.43 g ai / hm. 2 27.67g ai / hm 2 12.11g ai / hm 2 and 23.38g ai / hm 2 ; while for sensitive ryegrass, ED 50 The values ​​were 7.93 g ai / hm. 24.10g ai / hm 2 1.84g ai / hm 2 and 0.58g ai / hm 2 The relative resistance multiples were 11.91, 6.75, 6.58, and 40.31, respectively. This indicates that multiflora ryegrass exhibits multi-resistance to both ACCase inhibitor and ALS inhibitor herbicides, possibly due to the presence of genes contributing to this multi-resistance.

[0022] 1.2 Cloning and Expression Pattern Analysis of Resistance Genes The inventors used SMRT and RNA-Seq technologies to obtain the full-length sequence and complete transcript structure of *Lycium chinense*, and discovered... SCPL18 The gene is one of the NTSR (non-target-site resistance) genes in *Lysimachia christinae* related to the detoxification metabolism of clodinafop-propargyl and acesulfame potassium. qPCR validation revealed that... SCPL18 Gene expression trends are highly consistent with transcriptome data. Figure 1 ).

[0023] For research SCPL18 The inventors cloned the gene to investigate its function in the detoxification and metabolism of oxychlorpyrifos and chlorpyrifos. Using total RNA extracted from *Lysimachia christinae* as a template, and based on transcriptome data, they cloned the *Lysimachia christinae* gene. LmSCPL18 The EST sequence fragment of the gene was used to design two specific primers for PCR amplification based on the 3' and 5' end splicing sequences (see [link to primer]). Figure 2 a). The amplification primers are shown in Table 1, and the PCR reaction system is shown in Table 2.

[0024] Table 1 Amplification Primers 18-F ATGGAACACGCAGGACACCCAGC 18-R CAGCTCACAGAGCCTGCGTAC Table 2 PCR reaction system PrimeSTAR Max Premix (2X) 25μL 18-F 1μL 18-R 1μL template 0.5μL <![CDATA[ddH2O]]> 22.5μL The PCR program was as follows: 98℃ pre-denaturation for 5 min; 98℃ denaturation for 10 sec, 56℃ annealing for 15 sec, 72℃ extension for 8 sec, for a total of 34 cycles; and a final extension at 72℃ for 10 min.

[0025] The PCR amplification products were purified using a universal DNA purification and recovery kit (Tiangen) and ligated into the pMD (TaKaRa) vector. After transformation into DH5α competent E. coli cells, single clones were selected for identification (see...). Figure 2 b) Sequencing was performed, and finally, the flower ryegrass was successfully obtained. LmSCPL18 The full length of the gene.

[0026] 1.3. LmSCPL18 Construction of gene overexpression vectors Design specific primers according to Table 3 to clone and sequence the correct sequence. LmSCPL18 Using plasmids as templates, PrimeSTAR was used. ® PCR amplification was performed using Max DNA Polymerase (TaKaRa), and the PCR product was purified using a universal DNA purification and recovery kit to obtain the LmSCPL18 fragment with a pox homologous arm.

[0027] Table 3 Specific primers -F gtgttatacttctgcagggtaccATGGAACACGCAGGACACCCAGCT -R gttatcggatccataacgcgtCAGCTCACAGAGCCTGCGTAC The pox vector was double-digested with KpnI (FastDigest KpnI) and MluI (FastDigest MluI). The reaction mixture consisted of 2 μL 10X FastDigest Green Buffer, 1 μL FastDigest KpnI, 1 μL FastDigest MluI, 1000 ng pox plasmid, and sterile water to a final volume of 20 μL. The reaction conditions were 37°C for 5 min (see [link to reaction details]). Figure 3 a).

[0028] The reaction product was purified using a standard agarose gel DNA recovery kit to obtain a linearized vector. The purified target fragment and the linearized vector were then ligated using a homologous recombination kit (ClonExpress II One Step Cloning Kit). The ligation system consisted of: 0.03 pmol linearized vector, 0.06 pmol insert, 4 μL 5× CE II Buffer, 2 μL Exnase II, and ddH2O to a final volume of 20 μL. The reaction conditions were 37°C for 30 min.

[0029] 10 μL of the recombinant product was transduced into DH5α competent E. coli cells, and single clones were picked for sequencing identification. Figure 3 (b) to obtain the pox-LmSCPL18 fusion expression vector.

[0030] 1.4 Transformation of Agrobacterium with overexpression vector The plasmid obtained in the previous step was transformed into Agrobacterium EHA105 strain using an electroporation method, so that the overexpression vector could be transformed into rice callus for overexpression, in order to detect whether LmSCPL18 has herbicide resistance. The Agrobacterium transformation includes the following steps: a. Soak the electric shock cup and lid in 75% alcohol for 2 hours, then remove them in a clean bench and place them on clean filter paper to drain the water. After the alcohol has completely evaporated, insert them into ice to pre-cool. b. EHA105 Agrobacterium competent cells were thawed on ice; c. Pipette 1000 ng of recombinant plasmid pox-LmSCPL18 into EHA105 competent cells, gently aspirate and mix, then transfer it completely into an electroporation cup and cover the cup. d. Set the parameters of the electro-particle converter to 2.5kV and 5ms, wipe the water off the electro-particle cup, and quickly place it into the electro-particle tank for electro-particle discharge; d. Immediately insert the electric rotor into the ice after the electric shock; e. Add 700 μL of antibiotic-free LB medium, mix well by pipetting, and then transfer the entire mixture into a 1.5 mL centrifuge tube. Incubate at 28 °C with shaking for 2 h. f. Centrifuge at 4000 rpm for 1 min; g. Retain approximately 100 μL of supernatant, mix it thoroughly with the bacterial cells, and then spread it onto LB medium containing 50 μg / mL kanamycin and 20 μg / mL rifampin; h. Incubate in an inverted incubator at 28℃ for 2-3 days. Pick single colonies for colony PCR identification. Figure 3 c), positive transformants are available for use.

[0031] 1.5 Obtaining positive callus material from rice 1.5.1 Obtaining and transforming high-quality embryogenic callus Mature embryos of Nipponbare rice seeds were cultured for another 2-3 weeks. Vigorous, pale yellow, and densely granular embryogenic callus tissue was carefully selected in a clean bench. This callus was then transferred to fresh NB induction medium (see Table 4-8) and subcultured once for transformation. One to two days before transformation, *Agrobacterium glycerol pox-LmSCPL18* was streaked onto YEP solid plates containing 50 μg / mL Kan and 20 μg / mL Rif (the same below) for activation. Single colonies were then inoculated into YEP liquid medium containing Kan + Rif and cultured at 28°C with shaking at 200 rpm until OD... 600 The bacterial culture was centrifuged at 4°C and 5000 rpm for 10 min to collect the cells, and then resuspended in an equal volume of infection medium (see Table 9-13) to OD200. 600 Value 0.8, place on ice and wait for use.

[0032] Table 4. Formulations of NB induction medium, co-culture and screening medium in rice plant tissue culture. N6 Macroelements (20×) 50mL 50mL 50mL 50mL 50mL NB organic matter (200×) 5mL 5mL 5mL 5mL 5mL B5 Trace Elements (200×) 5mL 5mL 5mL 5mL 5mL Iron salts (100×) 10mL 10mL 10mL 10mL 10mL Plant gel 4.5g 4.5g 4.5g 4.5g 2.25g sucrose 30g 30g 30g 30g 30g Acid hydrolyzed casein 0.3g 0.3g 0.3g 2.0g — proline 2.875g — 2.875g 0.3g — Inositol — 1g — — — Sorbitol — — — 30g — 2,4-D (0.4 mg / mL) 5mL 5mL 5mL — — 100mM AS — 2mL — — — Hyg (50mg / mL) — — 1mL 1mL 1mL Termetidine (200 mg / mL) — — 1mL — — Kinetin (2.0 mg / mL) — — — 1mL — NAA (0.2 mg / mL) — — — 0.1mL — pH value 5.8 5.4 5.8 5.8 5.8 <![CDATA[ddH2O]]> Up to 1000mL Up to 1000mL Up to 1000mL Up to 1000mL Up to 1000mL Table 5. Formula for N6 macro-elements (20×, g / L) mother liquor <![CDATA[KNO3]]> 56.6g <![CDATA[(NH4)2SO4]]> 9.26g <![CDATA[KH2PO4]]> 8.00g <![CDATA[MgSO4·7H2O]]> 3.70g <![CDATA[CaC 12 ·H2O]]> 3.32g Table 6. NB Organic Matter (200×, g / L) Mother Liquor Formulation Inositol 56.6g Nicotinic acid (VB3) 0.1g VB6 (Pyridoxine HCl) 0.1g VB1 (Thiamine HCl) 0.2g glycine 0.4g Table 7. B5 Trace Element (200×, g / L) Mother Liquor Formula <![CDATA[MnSO4·4H2O]]> 56.6g Nicotinic acid (VB3) 0.1g VB6 (Pyridoxine HCl) 0.1g VB1 (Thiamine HCl) 0.2g glycine 0.4g Table 8. Formula for iron salt (100×, g / L) mother liquor <![CDATA[FeSO4·7H2O]]> 2.78g <![CDATA[Na2EDTA·2H2O]]> 3.73g Table 9. Formula for infection culture medium for rice plant tissue culture AA (20×) 100mL AA trace amount (200×) 5mL AA Organic (100×) 10mL Copper-copper masterbatch (2000×) 0.5mL sucrose 68.5g glucose 36g Acid hydrolyzed casein 0.5g glutamine 0.9g Aspartic acid 0.3g Arginine 0.1767g pH 5.2-5.3 <![CDATA[ddH2O]]> Up to 1000mL 100mM AS 2mL Table 10 Formula for AA Large Quantity (20×, g / L) Mother Liquor KCl 60g <![CDATA[NaH2PO4·2H2O]]> 3g <![CDATA[MgSO4·7H2O]]> 5g (244g anhydrous MgSO4) <![CDATA[CaCl2·2H2O]]> 3g (2.27g anhydrous CaCl2) Table 11 AA Micro-volume (200×, g / L) Mother Liquor Formulation <![CDATA[MgSO4·H2O]]> 1.52g <![CDATA[H3BO3]]> 0.6g <![CDATA[ZnSO4·7H2O]]> 0.4g Fe-EDTA 8g Table 12 Formula for AA Organic (100×, g / L) Mother Liquor Nicotinic acid (VB3) 0.1g VB6 (Pyridoxine HCl) 0.1g VB1 (Thiamine HCl) 1g Inositol 10g glycine 0.75g Table 13 Copper-copper mother liquor (2000×, g / L) formula <![CDATA[Na2MoO4·2H2O]]> 0.5g <![CDATA[CoC12·6H2O]]> 0.05g <![CDATA[CuSO4·5H2O]]> 0.05g KI 1.5g In a clean bench, place well-grown, loosely textured embryogenic callus tissue after subculture into a sterile Erlenmeyer flask or petri dish, and pour in the prepared Agrobacterium tumefaciens solution, ensuring that all callus tissue is submerged. Spread the infected callus tissue evenly on a co-culture medium (see Table 4-8) lined with sterile filter paper and incubate for 2-3 days.

[0033] 1.5.2 Screening of resistant callus After co-culture, the callus tissue was transferred to a sterile bottle containing sterile distilled water and gently shaken to wash 1-2 times. Then, it was washed 7 times with sterile water containing 200 mg / L termethin and 50 mg / L hygromycin. After blotting dry on filter paper, it was transferred to a selection medium containing 50 mg / L hygromycin and 200 mg / L termethin (Table 4-8) to screen for positive callus. The callus was cultured at 30°C in the dark. After 14 days, it was transferred to a new selection medium for subculture.

[0034] 1.5.3 Differentiation and Rooting of Resistant Plants After two rounds of selection, newly grown dense, firm, milky-yellow positive callus was selected and transferred to differentiation medium containing 50 mg / L hygromycin (see Table 4-8). The callus was first cultured in the dark for 3 days, then transferred to a 30℃ environment with a 16-hour light / 8-hour dark-light cycle. After approximately 2-4 weeks, the resistant callus began to differentiate into green buds and grow into seedlings. When the buds differentiated from the resistant callus reached approximately 2 cm in length, they were cut and transferred to rooting medium supplemented with 50 mg / L hygromycin (see Table 4-8) to induce rooting.

[0035] 1.5.4 Seedling hardening, transplanting, and molecular identification Select seedlings approximately 10cm tall with well-developed root systems, wash off the culture medium, and transplant them into soil in a greenhouse to obtain transformed plants. Use PCR to screen transgenic rice to detect whether the target gene has been integrated into the rice genome. Positive seedlings at the 3-4 leaf stage are selected. LmSCPL18 Transgenic rice lines were treated with foliar spraying. The spraying equipment used was a 3WP-2000 mobile spray tower from the Nanjing Institute of Mechanization, Ministry of Agriculture and Rural Affairs, with TP6501 fan-shaped nozzles. The spraying height was 300 mm, and the water consumption was 450 L / hm². 2 The experiment included two herbicides, cyclohexane and pyrazosulfuron, at a dosage of 45 g ai / hm. 2 and 10.75g ai / hm 2 Each treatment was repeated four times, with four plants per replicate. Mixtures of four plants from the same positive line were tested using qRT-PCR. LmSCPLl8 Regarding transcriptional expression, the experiment was conducted in quadruplicate, and the results are shown below. Figure 4 .

[0036] Depend on Figure 4 It can be seen that at a dosage of 45g ai / hm, the effect of oxychloride is... 2 Under the treatment, the expression level of transgenic rice overexpressing pox-LmSCPL18 was 8.18 times that of transgenic rice overexpressing pox-GFP; at a dose of 10.75 g ai / hm. 2 Under the treatment, the expression level of transgenic rice with pox-LmSCPL18 overexpression was 4.60 times that of transgenic rice with pox-GFP overexpression. (Note:) LmSCPL18 The expression level of the gene was significantly increased in pox-LmSCPL18 overexpressing transgenic rice, indicating that the gene has been successfully integrated into the rice genome.

[0037] 1.6 Determination of the sensitivity of transgenic rice to herbicides The sensitivity of different rice varieties to the ACCase inhibitor herbicide cyclophosphamide and the ALS inhibitor herbicide pyrazosulfuron was determined using a whole-plant bioassay.

[0038] The test material was a transfer LmSCPLl8 Genetically modified rice, with transgenic rice overexpressing pox-GFP as the control material, was used. Different rice materials were cultured until the 3-4 leaf stage, at which point foliar spraying was applied. The spraying equipment used was a 3WP-2000 mobile spray tower from the Nanjing Institute of Mechanization, Ministry of Agriculture and Rural Affairs, with TP6501 fan-shaped nozzles. The spray height was 300 mm, and the water consumption was 450 L / hm². 2The application rate of clopyralid is 45 g ai / hm. 2 The application rate of sulfadiazine was 10.75 g ai / hm. 2 Each treatment was repeated four times, with three transgenic rice plants per replicate. Fourteen days after application, the aboveground parts were cut off, and the average fresh weight per plant was measured. Fresh weight was used to evaluate the sensitivity of different materials to the herbicide. Results are shown below. Figure 5 .

[0039] Depend on Figure 5 It can be seen that at a dosage of 45g ai / hm, the effect of oxychloride is... 2 Under this treatment, the transgenic rice with pox-GFP overexpression died completely, while the transgenic rice with pox-LmSCPLl8 overexpression continued to grow at this dose. The fresh weight of the pox-LmSCPLl8 transgenic rice (1.71g) was 3.68 times that of the pox-GFP transgenic rice (0.46g).

[0040] At a dose of 10.75 g ai / hm 2 Under treatment, transgenic rice with pox-GFP overexpression showed wilting and yellowing symptoms, and its growth was inhibited, while transgenic rice with pox-LmSCPLl8 gene overexpression grew normally. The fresh weight of pox-LmSCPLl8 transgenic rice (1.53g) was 2.45 times that of pox-GFP transgenic rice (0.62g).

[0041] visible, LmSCPL18 Overexpression of the gene can induce significant resistance in transgenic rice to both oxychlorpyrifos and chlorpyrifos. Therefore, ryegrass... LmSCPL18 Genes can enhance plant resistance to herbicides and lay the foundation for breeding herbicide-resistant transgenic plants, which is of great significance.

[0042] The foregoing description of specific exemplary embodiments of the invention is for illustrative and explanatory purposes. These descriptions are not intended to limit the invention to the precise forms disclosed, and it will be apparent that many changes and variations can be made in accordance with the foregoing teachings. The exemplary embodiments were chosen and described in order to explain the specific principles of the invention and its practical application, thereby enabling those skilled in the art to implement and utilize various different exemplary embodiments of the invention, as well as various different choices and variations. The scope of the invention is intended to be defined by the claims and their equivalents.

Claims

1. A kind LmSCPL18 Genes, characterized by, The LmSCPL18 The nucleotide sequence of the gene is shown in SEQ ID NO.

1.

2. A protein polypeptide, characterized in that, The protein polypeptide is composed of the above LmSCPL18 It is encoded by a gene, and its amino acid sequence is shown in SEQ ID NO.2 of the sequence listing.

3. A gene expression vector, characterized in that, The gene expression vector comprises the one described in claim 1. LmSCPL18 Gene.

4. The claim 1 LmSCPL18 The application of genes, the protein polypeptides of claim 2, or the gene expression vectors of claim 3 in improving plant resistance to herbicides and in the breeding of herbicide-resistant transgenic plants.

5. The application according to claim 4, characterized in that, The herbicides include ACCase inhibitor herbicides and ALS inhibitor herbicides.

6. The application according to claim 5, characterized in that, The ACCase inhibitor herbicides include clodinafop-propargyl, quizalofop-p-ethyl, quizalofop-p-ethyl, quizalofop-p-ethyl, styrax, clethodim, and clethodim.

7. The application according to claim 5, characterized in that, The ALS inhibitor herbicides include mesosulfuron-methyl, pyrazosulfuron-methyl, fluazolidone, methoxyfenozide, imidacloprid, and methyl methoxyfenozide.

8. The application according to claim 5, characterized in that, The plants mentioned include wheat, barley, oats, rice, forage grass, corn, millet, and sorghum.

9. A method for improving plant resistance to herbicides, or for breeding herbicide-resistant transgenic plants, characterized in that, Specifically: LmSCPL18 The nucleic acid sequence of a gene is constructed into a gene expression vector, transformed into plants, and expressed in the plants to improve the herbicide resistance of plants or to achieve the breeding of herbicide-resistant transgenic plants; wherein, the... LmSCPL18 The nucleotide sequence of the gene is shown in SEQ ID NO.1 of the sequence listing.

10. The method according to claim 9, characterized in that, The herbicides include ACCase inhibitor herbicides and ALS inhibitor herbicides; The plants mentioned include wheat, barley, oats, rice, forage grass, corn, millet, rye, perennial ryegrass, stiff ryegrass, and sorghum.

Images