Breeding method of insect-resistant low-phenol high-quality cotton variety and intelligent breeding system

Abstract

The invention belongs to the technical field of cotton breeding, discloses a breeding method and an intelligent breeding system for an insect-resistant low-phenol high-quality cotton variety, and aims to remarkably improve the insect resistance of the cotton variety, reduce the use amount of chemical pesticides, relieve environmental pollution and protect the ecological environment. The content of phenolic substances in cotton is effectively reduced, the fiber quality and the textile processing performance are improved, and the requirement of the market for high-quality cotton is met. The breeding process is optimized, the breeding efficiency is improved, the breeding period is shortened, the breeding cost is reduced, and the sustainable development of the cotton industry is promoted. Through application of the intelligent breeding system, accurate management and analysis of breeding data are realized, and scientificity and accuracy of breeding decision are improved. Meanwhile, due to the intelligent and automatic breeding process, the labor cost is reduced, and the breeding efficiency is improved.

Description

Technical Field

[0001] The invention belongs to the technical field of cotton breeding, and in particular relates to a breeding method and an intelligent breeding system for an insect-resistant, low-phenol high-quality cotton variety. Background Art

[0002] With the development of modern agriculture, the demand for cotton varieties is becoming increasingly diversified, especially in terms of insect resistance, low phenol content and excellent fiber quality. Most of the existing cotton varieties in Xinjiang are not insect-resistant, and the use of chemical pesticides harms the ecological environment; the introduction of insect-resistant cotton varieties has a long planting period, poor insect resistance and poor fiber quality; the content of phenolic substances in cotton is high.

[0003] Through the above analysis, the problems and defects of the prior art are as follows:

[0004] Existing cotton varieties have poor insect resistance, the use of chemical pesticides harms the ecological environment; the quality of cotton fiber is poor; and the content of phenolic substances in cotton is high. Summary of the invention

[0005] In view of the problems existing in the prior art, the present invention provides a breeding method and an intelligent breeding system for insect-resistant, low-phenol high-quality cotton varieties.

[0006] The present invention is achieved in that a breeding method for insect-resistant, low-phenol high-quality cotton varieties comprises:

[0007] Step 1, screening and introduction of insect-resistant genes;

[0008] Screening for genes with high insect resistance;

[0009] Use transgenic technology to introduce insect-resistant genes into target organisms. Gene editing technologies such as CRISPR-Cas9 can be used to precisely edit or optimize the introduced insect-resistant genes to improve their expression efficiency or adjust their characteristics. Multi-generation screening and stable genetic testing;

[0010] Step 2, screening and cultivation of low-phenol genes;

[0011] Analysis of key genes for cotton phenol synthesis:

[0012] Screening for genotypes or mutants with low phenolic expression characteristics:

[0013] Low-phenol varieties are bred through hybridization and backcrossing.

[0014] Furthermore, the method for screening genes with high insect resistance is as follows:

[0015] (1) Using PCR polymerase chain reaction technology, amplify the insect-resistant gene fragment from cotton or other plant varieties known to be insect-resistant;

[0016] (2) Use bioinformatics tools to sequence gene fragments and predict the structure and function of the proteins they encode, thereby screening out genes with potential insect resistance;

[0017] (3) Under laboratory conditions, candidate genes are introduced into model plants, including Arabidopsis, through transgenic technology to observe their contribution to insect resistance.

[0018] Furthermore, the insect-resistant gene is introduced into the target organism using transgenic technology. Subsequently, gene editing technologies such as CRISPR-Cas9 can be used to precisely edit or optimize the introduced insect-resistant gene to improve its expression efficiency or adjust its characteristics.

[0019] 1) Design a specific CRISPRCas9 system, including a gRNA guide RNA specific for the target cotton genomic locus;

[0020] 2) constructing an expression vector containing an insect-resistant gene and a CRISPRCas9 system, and introducing the vector into cotton cells by Agrobacterium-mediated or gene gun methods;

[0021] 3) Under tissue culture conditions, screen the cell lines that have been successfully introduced and expressed, and further culture the regenerated plants.

[0022] Further, the multi-generation screening and stable genetic testing method:

[0023] In the regenerated plants, the presence and correct expression of the insect-resistant gene were verified by PCR and gene sequencing methods;

[0024] Conduct multiple generations of insect resistance testing under laboratory and field conditions to ensure stable inheritance and expression of insect resistance genes;

[0025] Screen out strains with stable insect resistance and clear genetic background for subsequent breeding work;

[0026] Further, the method for analyzing the key genes for the synthesis of cotton phenols:

[0027] Using genomics and transcriptomics technologies, we performed whole genome sequencing and RNA sequencing on cotton;

[0028] Through bioinformatics analysis, we found the gene families and regulatory networks related to the synthesis of phenolic substances;

[0029] Functional genomics methods including gene knockout and overexpression were used to verify the role of key genes in the synthesis of phenolic substances.

[0030] Furthermore, the method for screening genotypes or mutants with low phenol expression characteristics is as follows:

[0031] In natural populations or induced mutagenesis populations, individuals with low phenolic content are screened;

[0032] Through gene mapping and association analysis, we can find out the genes or genotypes associated with low phenol expression;

[0033] Breeding low-phenol varieties through hybridization and backcrossing:

[0034] The low-phenol genotype was crossed with high-quality cotton varieties, and through multiple generations of selection and self-pollination, the low-phenol gene was gradually made homozygous;

[0035] Using molecular marker-assisted selection technology to accelerate the aggregation and stabilization of low-phenol genes;

[0036] Through field trials, the stability of low-phenol varieties and the performance of other traits such as yield and quality are verified.

[0037] Another object of the present invention is to provide a breeding system for insect-resistant, low-phenol high-quality cotton varieties comprising:

[0038] Database construction module, used to establish cotton germplasm resource database:

[0039] Collect cotton germplasm resources, including seeds, DNA samples, and phenotypic data;

[0040] Using database management system, establish germplasm resource database including genetic information, phenotypic data, breeding history information;

[0041] Development module for developing intelligent breeding management software:

[0042] Use programming languages ​​and software development tools to develop intelligent breeding management software with data entry, query, analysis, and visualization functions;

[0043] Integrated automated data collection equipment including sensors and image recognition systems to achieve automated collection and processing of breeding data;

[0044] Mining and prediction module, used to perform data mining and predictive analysis using artificial intelligence algorithms:

[0045] Use machine learning and deep learning artificial intelligence algorithms to conduct in-depth mining and predictive analysis of breeding data;

[0046] Build prediction models to predict the performance of traits and key indicators of genetic stability of new varieties;

[0047] Through visualization technology, the analysis results are presented to users in the form of charts and reports;

[0048] Building blocks for constructing smart greenhouses and automated test fields:

[0049] Design and build smart greenhouses with environmental control, automated irrigation, fertilization, and pest and disease control functions;

[0050] In the field trials, automated monitoring equipment including drones and remote sensing equipment were introduced to achieve real-time monitoring and data collection of cotton growth status;

[0051] Through the construction of smart greenhouses and automated test fields, automated control and monitoring of the cotton breeding process can be achieved.

[0052] Another object of the present invention is to provide a computer device, which includes a memory and a processor, wherein the memory stores a computer program, and when the computer program is executed by the processor, the processor executes the steps of the breeding method for insect-resistant, low-phenol, high-quality cotton varieties.

[0053] Another object of the present invention is to provide a computer-readable storage medium storing a computer program, which, when executed by a processor, enables the processor to execute the steps of the breeding method for insect-resistant, low-phenol, high-quality cotton varieties.

[0054] Another object of the present invention is to provide an information data processing terminal, which is used to implement the breeding system of the insect-resistant, low-phenol, high-quality cotton variety.

[0055] In combination with the above technical solutions and the technical problems solved, the advantages and positive effects of the technical solutions to be protected by the present invention are as follows:

[0056] First, in view of the technical problems existing in the above-mentioned prior art, some creative technical effects are brought about after solving the problems. The specific description is as follows:

[0057] Significantly improve the insect resistance of cotton varieties, reduce the use of chemical pesticides, reduce environmental pollution, and protect the ecological environment.

[0058] Effectively reduce the content of phenolic substances in cotton, improve fiber quality and textile processing performance, and meet the market demand for high-quality cotton.

[0059] Optimize the breeding process, improve breeding efficiency, shorten the breeding cycle, reduce breeding costs, and promote the sustainable development of the cotton industry.

[0060] Through the application of intelligent breeding system, accurate management and analysis of breeding data can be achieved, and the scientificity and accuracy of breeding decisions can be improved. At the same time, the intelligent and automated breeding process reduces labor costs and improves breeding efficiency.

[0061] This technical solution solves the key problems in cotton breeding through the screening and introduction of insect-resistant genes, the screening and cultivation of low-phenol genes, and the construction of an intelligent breeding system, and realizes the cultivation of insect-resistant, low-phenol high-quality cotton varieties. The implementation of this technical solution will bring significant ecological, economic and social benefits, and is of great significance to promoting the green development and high-quality development of the cotton industry.

[0062] Second, the present invention solves the technical difficulties in the existing technology in terms of insect resistance and low phenolic properties of cotton varieties. Traditional cotton breeding methods are difficult to breed cotton varieties with multiple excellent traits in a short period of time due to their long cycle and low efficiency. However, the present invention uses modern technologies such as molecular marker-assisted selection, gene editing and high-throughput genome sequencing to quickly and accurately screen out key genes related to insect resistance and low phenolic properties, thereby significantly shortening the breeding cycle and improving breeding efficiency.

[0063] In terms of insect resistance improvement, the present invention uses CRISPR-Cas9 gene editing technology to accurately introduce highly effective insect-resistant genes such as Bt genes into the cotton genome. The problems of low gene integration efficiency and poor stability in traditional methods have been effectively solved. Through multi-generation screening and stable genetic testing, the cotton varieties bred by the present invention showed significant insect resistance in field trials, and the pest loss rate was reduced by more than 70%, which significantly improved the insect resistance of cotton, reduced the use of pesticides, and reduced production costs and environmental pollution.

[0064] In terms of improving low-phenol characteristics, the present invention uses high-throughput genome sequencing and bioinformatics analysis to accurately screen out key genes related to the synthesis of phenolic substances, such as the PAL gene. By reducing its expression level through gene editing technology, low-phenol cotton varieties were successfully bred. These varieties not only have a phenolic substance content reduced by more than 50%, but also maintain high-yield and high-quality fiber characteristics. Compared with traditional breeding methods, the present invention has made significant progress in improving low-phenol characteristics and enhanced the market competitiveness of cotton products.

[0065] The present invention successfully bred insect-resistant, low-phenol, high-quality cotton varieties through scientific and reasonable gene screening, introduction and breeding methods, showing significant technological progress in insect resistance, low-phenol characteristics, fiber quality and breeding efficiency. These technological innovations not only improve the overall performance of cotton varieties, but also promote the development of cotton breeding technology, and have broad application prospects and important economic value. BRIEF DESCRIPTION OF THE DRAWINGS

[0066] Figure 1 It is a flow chart of the breeding method of insect-resistant, low-phenol high-quality cotton varieties provided by an embodiment of the present invention.

[0067] Figure 2It is a flow chart of the method for screening genes with high insect resistance provided by an embodiment of the present invention.

[0068] Figure 3 First, the insect-resistant gene is introduced into the target organism using transgenic technology. Subsequently, the introduced insect-resistant gene can be precisely edited or optimized using gene editing technologies such as CRISPR-Cas9 to improve its expression efficiency or adjust its characteristics. Flowchart.

[0069] Figure 4 It is a structural block diagram of the breeding system for insect-resistant, low-phenol high-quality cotton varieties provided by an embodiment of the present invention. DETAILED DESCRIPTION

[0070] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0071] like Figure 1 As shown, a breeding method for an insect-resistant, low-phenol high-quality cotton variety provided by an embodiment of the present invention comprises the following steps:

[0072] S101, screening and introduction of insect-resistant genes;

[0073] Screening for genes with high insect resistance;

[0074] Use gene editing technologies including CRISPRCas9 to introduce insect resistance genes;

[0075] multigenerational screening and stable genetic testing;

[0076] S102, screening and cultivation of low-phenol genes;

[0077] Analysis of key genes for cotton phenol synthesis:

[0078] Screening for genotypes or mutants with low phenolic expression characteristics:

[0079] Low-phenol varieties are bred through hybridization and backcrossing.

[0080] like Figure 2 As shown, the method for screening genes with high insect resistance provided by the embodiment of the present invention is:

[0081] S201, an insect-resistant gene fragment amplified from cotton or other plant varieties known to be insect-resistant using PCR polymerase chain reaction technology;

[0082] S202, using bioinformatics tools to sequence gene fragments and predict the structure and function of the proteins they encode, thereby screening out genes with potential insect resistance;

[0083] S203, under laboratory conditions, candidate genes are introduced into model plants including Arabidopsis thaliana through transgenic technology to observe their contribution to insect resistance.

[0084] like Figure 3 As shown, the embodiment of the present invention firstly uses transgenic technology to introduce the insect-resistant gene into the target organism. Subsequently, gene editing technology such as CRISPR-Cas9 can be used to precisely edit or optimize the introduced insect-resistant gene to improve its expression efficiency or adjust its characteristics. :

[0085] S301, designing a specific CRISPRCas9 system, including a gRNA guide RNA specific for the target cotton genomic locus;

[0086] S302, constructing an expression vector containing an insect-resistant gene and a CRISPRCas9 system, and introducing the vector into cotton cells by Agrobacterium-mediated method or gene gun method;

[0087] S303, under tissue culture conditions, screen the cell lines that have been successfully introduced and expressed, and further culture the regenerated plants.

[0088] The multi-generation screening and stable genetic testing method provided by the embodiment of the present invention:

[0089] In the regenerated plants, the presence and correct expression of the insect-resistant gene were verified by PCR and gene sequencing methods;

[0090] Conduct multiple generations of insect resistance testing under laboratory and field conditions to ensure stable inheritance and expression of insect resistance genes;

[0091] Screen out strains with stable insect resistance and clear genetic background for subsequent breeding work;

[0092] The key gene method for analyzing the synthesis of cotton phenols provided in the embodiment of the present invention is as follows:

[0093] Using genomics and transcriptomics technologies, we performed whole genome sequencing and RNA sequencing on cotton;

[0094] Through bioinformatics analysis, we found the gene families and regulatory networks related to the synthesis of phenolic substances;

[0095] Functional genomics methods including gene knockout and overexpression were used to verify the role of key genes in the synthesis of phenolic substances.

[0096] The method for screening genotypes or mutants with low phenol expression characteristics provided in the embodiments of the present invention is as follows:

[0097] In natural populations or induced mutagenesis populations, individuals with low phenolic content are screened;

[0098] Through gene mapping and association analysis, we can find out the genes or genotypes associated with low phenol expression;

[0099] Breeding low-phenol varieties through hybridization and backcrossing:

[0100] The low-phenol genotype was crossed with high-quality cotton varieties, and through multiple generations of selection and self-pollination, the low-phenol gene was gradually made homozygous;

[0101] Using molecular marker-assisted selection technology to accelerate the aggregation and stabilization of low-phenol genes;

[0102] Through field trials, the stability of low-phenol varieties and the performance of other traits such as yield and quality are verified.

[0103] like Figure 4 As shown, a breeding system for insect-resistant, low-phenol high-quality cotton varieties provided by an embodiment of the present invention comprises:

[0104] Database construction module, used to establish cotton germplasm resource database:

[0105] Collect cotton germplasm resources, including seeds, DNA samples, and phenotypic data;

[0106] Using database management system, establish germplasm resource database including genetic information, phenotypic data, breeding history information;

[0107] Development module for developing intelligent breeding management software:

[0108] Use programming languages ​​and software development tools to develop intelligent breeding management software with data entry, query, analysis, and visualization functions;

[0109] Integrated automated data collection equipment including sensors and image recognition systems to achieve automated collection and processing of breeding data;

[0110] Mining and prediction module, used to perform data mining and predictive analysis using artificial intelligence algorithms:

[0111] Use machine learning and deep learning artificial intelligence algorithms to conduct in-depth mining and predictive analysis of breeding data;

[0112] Build prediction models to predict the performance of traits and key indicators of genetic stability of new varieties;

[0113] Through visualization technology, the analysis results are presented to users in the form of charts and reports;

[0114] Building blocks for constructing smart greenhouses and automated test fields:

[0115] Design and build smart greenhouses with environmental control, automated irrigation, fertilization, and pest and disease control functions;

[0116] In the field trials, automated monitoring equipment including drones and remote sensing equipment were introduced to achieve real-time monitoring and data collection of cotton growth status;

[0117] Through the construction of smart greenhouses and automated test fields, automated control and monitoring of the cotton breeding process can be achieved.

[0118] Another object of the present invention is to provide a computer device, which includes a memory and a processor, wherein the memory stores a computer program, and when the computer program is executed by the processor, the processor executes the steps of the breeding method for insect-resistant, low-phenol, high-quality cotton varieties.

[0119] Another object of the present invention is to provide a computer-readable storage medium storing a computer program, which, when executed by a processor, enables the processor to execute the steps of the breeding method for insect-resistant, low-phenol, high-quality cotton varieties.

[0120] Another object of the present invention is to provide an information data processing terminal, which is used to implement the breeding system of the insect-resistant, low-phenol, high-quality cotton variety.

[0121] The present invention is specifically implemented:

[0122] Detailed description of the specific implementation methods of each part:

[0123] 1. Screening and introduction of insect-resistant genes

[0124] 1.1. Screening for genes with high insect resistance:

[0125] Using PCR (polymerase chain reaction) technology, insect-resistant gene fragments are amplified from cotton or other plant varieties known to be insect-resistant.

[0126] Bioinformatics tools are used to perform sequence analysis on gene fragments and predict the structure and function of the proteins they encode, thereby screening out genes with potential insect resistance.

[0127] Under laboratory conditions, candidate genes are introduced into model plants (such as Arabidopsis thaliana) through transgenic technology to observe their contribution to insect resistance.

[0128] 1.2. Use gene editing technology (such as CRISPRCas9) to introduce insect-resistant genes:

[0129] A specific CRISPRCas9 system was designed, including a specific gRNA (guide RNA) targeting the target cotton genomic locus.

[0130] An expression vector containing an insect-resistant gene and a CRISPRCas9 system is constructed, and the vector is introduced into cotton cells through Agrobacterium-mediated or gene gun methods.

[0131] Under tissue culture conditions, cell lines that have been successfully introduced and expressed are screened and further regenerated plant cultures are performed.

[0132] 1.3. Multi-generation screening and stable genetic testing:

[0133] In the regenerated plants, the presence and correct expression of insect-resistant genes were verified by PCR, gene sequencing and other methods.

[0134] Insect resistance is tested over multiple generations under laboratory and field conditions to ensure stable inheritance and expression of insect-resistant genes.

[0135] Screen out strains with stable insect resistance and clear genetic background for subsequent breeding work.

[0136] 2. Screening and cultivation of low-phenol genes

[0137] 2.1. Analysis of key genes for cotton phenol synthesis:

[0138] Using genomics and transcriptomics technologies, whole genome sequencing and RNA sequencing of cotton were performed.

[0139] Through bioinformatics analysis, the gene families and regulatory networks related to the synthesis of phenolic substances were identified.

[0140] Through functional genomics methods (such as gene knockout, overexpression, etc.), the role of key genes in the synthesis of phenolic substances is verified.

[0141] 2.2. Screening of genotypes or mutants with low phenol expression characteristics:

[0142] In natural or mutagenic populations, screen for individuals with low phenolic content.

[0143] Through gene mapping and association analysis, genes or genotypes associated with low phenol expression were identified.

[0144] 2.3. Cultivate low-phenol varieties through hybridization, backcrossing, etc.:

[0145] The low-phenol genotype was crossed with high-quality cotton varieties, and through multiple generations of selection and self-pollination, the low-phenol gene was gradually made homozygous.

[0146] Molecular marker-assisted selection technology was used to accelerate the aggregation and stabilization of low-phenol genes.

[0147] Through field trials, the stability of low-phenol varieties and the performance of other traits such as yield and quality are verified.

[0148] 3. Construction of intelligent breeding system

[0149] 3.1. Establish a cotton germplasm resource database:

[0150] Collect cotton germplasm resources, including seeds, DNA samples, phenotypic data, etc.

[0151] Using the database management system, a germplasm resource database including genetic information, phenotypic data, breeding history and other information is established.

[0152] 3.2. Develop intelligent breeding management software:

[0153] Use programming languages ​​and software development tools to develop intelligent breeding management software with functions such as data entry, query, analysis, and visualization.

[0154] Integrate automated data acquisition equipment (such as sensors, image recognition systems, etc.) to realize automated collection and processing of breeding data.

[0155] 3.3. Using artificial intelligence algorithms for data mining and predictive analysis:

[0156] Use artificial intelligence algorithms such as machine learning and deep learning to conduct in-depth mining and predictive analysis of breeding data.

[0157] Build prediction models to predict key indicators such as trait performance and genetic stability of new varieties.

[0158] Through visualization technology, the analysis results are presented to users in the form of charts, reports, etc.

[0159] 3.4. Construction of smart greenhouse and automated test field:

[0160] Design and build smart greenhouses with functions such as environmental control, automated irrigation, fertilization, and pest and disease control.

[0161] In field trials, automated monitoring equipment (such as drones, remote sensing equipment, etc.) was introduced to achieve real-time monitoring and data collection of cotton growth status.

[0162] Through the construction of smart greenhouses and automated test fields, automated control and monitoring of the cotton breeding process can be achieved.

[0163] Technical effects

[0164] 1. Significantly improve the insect resistance of cotton varieties, reduce the use of chemical pesticides, reduce environmental pollution, and protect the ecological environment.

[0165] 2. Effectively reduce the content of phenolic substances in cotton, improve fiber quality and textile processing performance, and meet the market demand for high-quality cotton.

[0166] 3. Optimize the breeding process, improve breeding efficiency, shorten the breeding cycle, reduce breeding costs, and promote the sustainable development of the cotton industry.

[0167] 4. Through the application of intelligent breeding system, accurate management and analysis of breeding data can be achieved, and the scientificity and accuracy of breeding decisions can be improved. At the same time, the intelligent and automated breeding process reduces labor costs and improves breeding efficiency.

[0168] This technical solution solves the key problems in cotton breeding through the screening and introduction of insect-resistant genes, the screening and cultivation of low-phenol genes, and the construction of an intelligent breeding system, and realizes the cultivation of insect-resistant, low-phenol high-quality cotton varieties. The implementation of this technical solution will bring significant ecological, economic and social benefits, and is of great significance to promoting the green development and high-quality development of the cotton industry.

[0169] The present invention proposes a breeding method for insect-resistant, low-phenol high-quality cotton varieties, which involves the screening and introduction of insect-resistant genes, as well as the screening and cultivation of low-phenol genes. The following is a detailed working principle:

[0170] Step 1: Screening and introduction of insect-resistant genes

[0171] 1) Screening of insect-resistant genes:

[0172] First, bioinformatics tools and genomic data analysis are used to screen out highly efficient genes related to insect resistance, which usually come from plants or other organisms with natural insect resistance.

[0173] The insect-resistant effects of these genes are verified through experiments, and genes with significant insect-resistant effects are selected for subsequent operations.

[0174] 2) Gene introduction:

[0175] Using gene editing technologies such as CRISPR-Cas9, the selected insect-resistant genes are precisely introduced into the cotton genome. CRISPR-Cas9 technology can edit at specific gene sites, thus ensuring that the insect-resistant genes are integrated and expressed at the target location.

[0176] In order to ensure the success of gene editing, molecular biological methods such as PCR and Southern blot were used to verify the integration and expression level of insect-resistant genes in the cotton genome.

[0177] 3) Multi-generation screening and stable genetic testing:

[0178] The gene-edited cotton is planted and screened for multiple generations to select plants with stable insect resistance. Through continuous screening for several generations, it is ensured that the insect-resistant gene can be stably inherited and maintain its high insect resistance.

[0179] In each generation, the performance and stability of insect-resistant genes are evaluated through field trials and insect resistance testing, and ultimately highly insect-resistant cotton varieties are selected.

[0180] Step 2: Screening and cultivation of low-phenol genes

[0181] 1) Analysis of key genes for cotton phenol synthesis:

[0182] Through genomic analysis and bioinformatics tools, key genes involved in the synthesis of phenolic substances in cotton were identified. The expression levels of these genes directly affect the content of phenolic substances in cotton.

[0183] Through functional verification, we determined which genes played a key role in the synthesis of phenolic substances.

[0184] 2) Screening for genotypes or mutants with low phenol expression characteristics:

[0185] Among the existing cotton varieties, genotypes with low phenol expression characteristics are found through mutant screening or natural variation screening. These genotypes show lower accumulation of phenolic substances in the phenolic substance biosynthesis pathway.

[0186] Gene editing technology (such as CRISPR-Cas9) or conventional mutagenesis technology is used to induce the cotton genome to produce low phenol expression mutants.

[0187] 3) Cultivate low-phenol varieties through hybridization and backcrossing:

[0188] By hybridizing the low-phenol expression genotype with high-yield and high-quality cotton varieties and combining the excellent traits, a cotton variety with both low-phenol characteristics and high yield and high quality was bred.

[0189] Through multiple generations of backcrossing, the low-phenol characteristics are stabilized and the excellent traits of the target varieties are retained. In each generation, the performance and stability of the low-phenol characteristics are evaluated through chemical analysis and field trials, and finally a low-phenol high-quality cotton variety is bred.

[0190] Specific examples:

[0191] 1) Examples of insect-resistant gene screening and introduction:

[0192] The Bt gene was selected from plants with natural insect resistance (such as Arabidopsis thaliana) and introduced into the cotton genome using CRISPR-Cas9 technology. In multiple generations of screening, more than 80% of cotton plants showed high insect resistance.

[0193] 2) Examples of low-phenol gene screening and cultivation:

[0194] Through genome analysis, the PAL gene related to the synthesis of phenolic substances in cotton was identified. The PAL gene was edited using CRISPR-Cas9 technology to screen out cotton mutants with low phenol expression. After multiple generations of hybridization and backcrossing, a low-phenol high-quality cotton variety was stably obtained, and its phenolic content was reduced by more than 50%.

[0195] In summary, the present invention successfully bred insect-resistant, low-phenol, and high-quality cotton varieties through scientific and reasonable gene screening, introduction, and breeding methods, which showed significant improvements and enhancements in insect resistance and fiber quality, and has important application value and market prospects.

[0196] Step 1: Screening and introduction of insect-resistant genes

[0197] 1) Screening of insect-resistant genes:

[0198] The Bt gene was screened from plants with natural insect resistance (such as Arabidopsis thaliana) and is known to be highly effective in killing a variety of pests.

[0199] Bioinformatics tools and genomic data analysis are used to determine the appropriate integration site of the Bt gene in the cotton genome, and experimental verification is carried out to select genes with significant insect resistance effects.

[0200] 2) Gene introduction:

[0201] Using CRISPR-Cas9 gene editing technology, the Bt gene is precisely introduced into the target site of the cotton genome.

[0202] The integration and expression level of the Bt gene in the cotton genome were verified by PCR and Southern blot technology to ensure successful integration and efficient expression of the gene.

[0203] 3) Multi-generation screening and stable genetic testing:

[0204] The gene-edited cotton plants are planted and screened for multiple generations to select plants with stably inherited insect resistance.

[0205] In each generation, the performance of insect-resistant genes is evaluated through field trials and insect resistance testing, and finally highly insect-resistant cotton varieties are bred. Experimental data show that in field trials, the pest loss rate of insect-resistant cotton varieties was reduced by more than 70%.

[0206] Example 2: Breeding of low-phenol high-quality cotton varieties

[0207] Step 2: Screening and cultivation of low-phenol genes

[0208] 1) Analysis of key genes for cotton phenol synthesis:

[0209] Through genome analysis and bioinformatics tools, the key gene PAL (phenylalanine ammonia lyase) involved in the synthesis of phenolic substances in cotton was identified.

[0210] Gene expression analysis and functional verification confirmed that the PAL gene plays a key role in the synthesis of phenolic substances.

[0211] 2) Screening for genotypes or mutants with low phenol expression characteristics:

[0212] Among existing cotton varieties, genotypes with low phenol expression characteristics are found through mutant screening or natural variation screening.

[0213] The PAL gene was edited using CRISPR-Cas9 technology to generate a cotton mutant with low phenol expression, and chemical analysis verified that its phenolic content was significantly reduced.

[0214] 3) Cultivate low-phenol varieties through hybridization and backcrossing:

[0215] By hybridizing the cotton genotype with low phenol expression with the high-yield and high-quality cotton varieties and combining the excellent traits, a cotton variety with both low phenol characteristics and high yield and high quality was bred.

[0216] Through multiple generations of backcrossing, the low-phenol characteristics are stabilized and the excellent traits of the target varieties are retained. In each generation, the performance and stability of the low-phenol characteristics are evaluated through chemical analysis and field trials. Experimental data show that the phenolic content of the selected low-phenol cotton varieties is reduced by more than 50%, while maintaining high yield and high-quality fiber characteristics.

[0217] These two embodiments demonstrate the breeding method of the present invention for insect resistance and low phenol characteristics and its successful application. Through scientific gene screening, introduction and multi-generation screening, the insect resistance and fiber quality of cotton varieties are significantly improved, which has important application value and market prospects.

[0218] The breeding method of the insect-resistant, low-phenol high-quality cotton variety of the present invention has achieved remarkable positive effects in the process of research and development and use, and has obvious advantages over the prior art. The following content is described in detail in combination with the data, tables, etc. of the test process:

[0219] First, by screening and introducing highly efficient insect-resistant genes, the present invention significantly improves the insect resistance of cotton. During the experiment, the CRISPR-Cas9 gene editing technology was used to accurately introduce the Bt gene into the cotton genome. Multi-generation screening and stable genetic testing showed that the pest loss rate of cotton varieties introduced with the Bt gene in field trials was reduced by more than 70%, and insect resistance was improved compared with traditional methods. In addition, the efficient integration and stable expression of the Bt gene enable these cotton varieties to show significant resistance to a variety of pests, reduce the use of pesticides, and reduce environmental pollution and planting costs.

[0220] Secondly, in terms of low-phenol gene screening and cultivation, the present invention identified the key gene PAL related to the synthesis of phenolic substances through high-throughput genome sequencing and bioinformatics analysis. The PAL gene was edited by CRISPR-Cas9 technology to screen out cotton mutants expressing low phenol. Experimental data showed that the phenolic content of the selected low-phenol cotton varieties was reduced by more than 50%, while maintaining high-yield and high-quality fiber characteristics. In field trials, these low-phenol cotton varieties showed higher economic value and market competitiveness.

[0221] In addition, the method of the present invention significantly shortens the breeding cycle and improves breeding efficiency. Traditional cotton breeding methods have long cycles and low efficiency, while the present invention uses modern breeding technologies such as molecular marker-assisted selection, gene introduction and high-throughput genome sequencing to quickly screen and cultivate cotton varieties with excellent traits. Test data show that through the method of the present invention, the breeding cycle is shortened by about 30%, the breeding efficiency is increased by more than 50%, and the promotion and application of high-quality cotton varieties is greatly accelerated.

[0222] Case study on the implementation of breeding methods for insect-resistant, low-phenolic, high-quality cotton varieties.

[0223] 1. Screening and introduction of insect-resistant genes

[0224] 1. From known biological resources with high insect resistance, use molecular biology technology to screen out genes with significant insect resistance, such as insect resistance protein genes isolated from certain insect pathogens such as Bacillus thuringiensis (Bt).

[0225] 2. Use transgenic technology, such as Agrobacterium-mediated method or gene gun method, to introduce the selected insect-resistant gene into the genome of cotton cells. This step needs to be carried out under laboratory conditions and ensure the stable integration of the gene.

[0226] 3. Through tissue culture technology, the transgenic cells are cultivated into complete cotton plants.

[0227] 2. Gene Editing Using CRISPR-Cas9

[0228] 1. Design a specific CRISPR-Cas9 system that can specifically recognize and edit the introduced insect resistance gene. This can improve the expression efficiency of the gene or fine-tune the gene to adapt to the growth environment of cotton.

[0229] 2. Introduce the CRISPR-Cas9 system into transgenic cotton plants through appropriate transformation methods.

[0230] 3. Screen the edited cotton plants and select those with the best insect resistance and the most stable gene editing effects.

[0231] 3. Stable genetic testing of insect resistance

[0232] 1. Propagate gene-edited cotton plants over multiple generations to observe the genetic stability of their insect resistance.

[0233] 2. Evaluate the resistance of cotton plants to different pests through field trials and laboratory tests to ensure the durability and effectiveness of insect resistance.

[0234] 4. Screening and cultivation of low-phenol genes

[0235] 1. Use gene sequencing and analysis technology to identify the key genes responsible for the synthesis of phenolic substances in cotton.

[0236] 2. Screen out genotypes or mutants with naturally low phenolic properties from existing cotton germplasm resources. These resources come from traditional breeding programs or natural mutations.

[0237] 3. Introducing low-phenol traits into cotton varieties that already have insect resistance through hybridization and backcrossing techniques. This step requires careful selection of parents and tracking of the genetic traits of the offspring.

[0238] 4. After many generations of screening and backcrossing, a high-quality cotton variety with both stable insect resistance and low phenol properties was finally bred.

[0239] V. Field trials and variety promotion

[0240] 1. Conduct field trials in different ecological regions to evaluate the adaptability, yield and fiber quality of new varieties.

[0241] 2. Collect feedback from farmers and textile companies to continuously improve and optimize new varieties.

[0242] 3. Once a new variety demonstrates stable high-quality characteristics, it can be planted and promoted on a large scale through seed companies or agricultural extension agencies.

[0243] Through the above steps, a high-quality cotton variety that is both insect-resistant and low in phenol can be successfully bred, bringing practical economic and ecological benefits to agricultural production.

[0244] The following table shows the comparative data of the insect-resistant, low-phenol high-quality cotton varieties bred by the present invention and traditional varieties in terms of insect resistance, low-phenol characteristics and fiber quality:

[0245]

[0246] In summary, the present invention shows significant technical progress in insect resistance, low phenolic properties, fiber quality and breeding efficiency, and shows broad application prospects. This innovative breeding method not only provides scientific and effective technical means for the cotton industry, but also has important significance for improving cotton yield and quality, reducing production costs and promoting sustainable agricultural development.

[0247] The above description is only a specific implementation mode of the present invention, but the protection scope of the present invention is not limited thereto. Any modification, equivalent substitution and improvement made by any technician familiar with the technical field within the technical scope disclosed by the present invention and within the spirit and principle of the present invention should be covered by the protection scope of the present invention.

Claims

1. A method for breeding insect-resistant, low-phenol high-quality cotton varieties, characterized in that: The following steps are involved: Screening and introduction of insect-resistant genes: Screening of genes with high insect resistance; The insect-resistant gene is introduced into the target organism using transgenic technology; the introduced insect-resistant gene can be precisely edited or optimized using gene editing technologies such as CRISPR-Cas9 to improve its expression efficiency or adjust its characteristics; Through multi-generation screening and stable genetic testing, cotton plants with stable insect resistance are selected; Screening and breeding of low-phenol genes: Analyze the key genes for the synthesis of cotton phenolic substances; screen genotypes or mutants with low-phenol expression characteristics; cultivate low-phenol varieties through hybridization and backcrossing, and stabilize the genetic low-phenol characteristics through multiple generations of screening and backcrossing.

2. The breeding method of insect-resistant, low-phenol high-quality cotton varieties according to claim 1, characterized in that: The gene editing technique comprises the following steps: Determine the appropriate integration site of insect resistance genes in the cotton genome through bioinformatics tools and genomic data analysis; Use CRISPR-Cas9 technology to precisely introduce insect-resistant genes into the target site of the cotton genome; The integration and expression levels of insect-resistant genes in the cotton genome were verified by PCR and Southern blot techniques; When screening low-phenol genotypes or mutants, gene expression analysis and functional verification are used to confirm the effects of key genes on the synthesis of phenolic substances, and the genetic low-phenol characteristics are stabilized through multiple generations of hybridization and backcrossing.

3. The breeding method of insect-resistant, low-phenol high-quality cotton varieties as claimed in claim 1, characterized in that: The method for screening genes with high insect resistance: (1) Using PCR polymerase chain reaction technology, amplify the insect-resistant gene fragment from cotton or other plant varieties known to be insect-resistant; (2) Use bioinformatics tools to sequence gene fragments and predict the structure and function of the proteins they encode, thereby screening out genes with potential insect resistance; (3) Under laboratory conditions, candidate genes are introduced into model plants, including Arabidopsis, through transgenic technology to observe their contribution to insect resistance.

4. The method for breeding insect-resistant, low-phenol high-quality cotton varieties as claimed in claim 1, characterized in that: First, the insect-resistant gene is introduced into the target organism using transgenic technology. Subsequently, gene editing technologies such as CRISPR-Cas9 can be used to precisely edit or optimize the introduced insect-resistant gene to improve its expression efficiency or adjust its characteristics. 1) Design a specific CRISPRCas9 system, including a gRNA guide RNA specific for the target cotton genomic locus; 2) constructing an expression vector containing an insect-resistant gene and a CRISPRCas9 system, and introducing the vector into cotton cells by Agrobacterium-mediated or gene gun methods; 3) Under tissue culture conditions, screen the cell lines that have been successfully introduced and expressed, and further culture the regenerated plants.

5. The method for breeding insect-resistant, low-phenol high-quality cotton varieties as claimed in claim 1, characterized in that: The multi-generation screening and stable genetic testing method: In the regenerated plants, the presence and correct expression of the insect-resistant gene were verified by PCR and gene sequencing methods; Conduct multiple generations of insect resistance testing under laboratory and field conditions to ensure stable inheritance and expression of insect resistance genes; Screen out strains with stable insect resistance and clear genetic background for subsequent breeding work.

6. The method for breeding insect-resistant, low-phenol high-quality cotton varieties as claimed in claim 1, characterized in that: The method for analyzing the key genes for the synthesis of cotton phenols: Using genomics and transcriptomics technologies, whole genome sequencing and RNA sequencing of cotton were performed; Through bioinformatics analysis, we found the gene families and regulatory networks related to the synthesis of phenolic substances; Functional genomics methods including gene knockout and overexpression were used to verify the role of key genes in the synthesis of phenolic substances.

7. The method for breeding insect-resistant, low-phenol high-quality cotton varieties as claimed in claim 1, characterized in that: The method for screening genotypes or mutants with low phenol expression characteristics: In natural populations or induced mutagenesis populations, individuals with low phenolic content are screened; Through gene mapping and association analysis, we can find out the genes or genotypes associated with low phenol expression; Breeding low-phenol varieties through hybridization and backcrossing: The low-phenol genotype was crossed with high-quality cotton varieties, and through multiple generations of selection and self-pollination, the low-phenol gene was gradually made homozygous; Using molecular marker-assisted selection technology to accelerate the aggregation and stabilization of low-phenol genes; Through field trials, the stability of low-phenol varieties and the performance of other traits such as yield and quality are verified.

8. A breeding system for insect-resistant, low-phenol high-quality cotton varieties for implementing the breeding method for insect-resistant, low-phenol high-quality cotton varieties according to any one of claims 1 to 7, characterized in that: The breeding system of the insect-resistant, low-phenol high-quality cotton variety comprises: Database construction module, used to establish cotton germplasm resource database: Collect cotton germplasm resources, including seeds, DNA samples, and phenotypic data; Using database management system, establish germplasm resource database including genetic information, phenotypic data, breeding history information; Development module for developing intelligent breeding management software: Use programming languages ​​and software development tools to develop intelligent breeding management software with data entry, query, analysis, and visualization functions; Integrated automated data collection equipment including sensors and image recognition systems to achieve automated collection and processing of breeding data; Mining and prediction module, used to perform data mining and predictive analysis using artificial intelligence algorithms: Use machine learning and deep learning artificial intelligence algorithms to conduct in-depth mining and predictive analysis of breeding data; Build prediction models to predict the performance of traits and key indicators of genetic stability of new varieties; Through visualization technology, the analysis results are presented to users in the form of charts and reports; Building blocks for constructing smart greenhouses and automated test fields: Design and build smart greenhouses with environmental control, automated irrigation, fertilization, and pest and disease control functions; In the field trials, automated monitoring equipment including drones and remote sensing equipment were introduced to achieve real-time monitoring and data collection of cotton growth status; Through the construction of smart greenhouses and automated test fields, automated control and monitoring of the cotton breeding process can be achieved.

9. A method for breeding insect-resistant, low-phenol high-quality cotton varieties, characterized in that: The method comprises the following steps: a. Screening and introduction of insect-resistant genes, including: Screening for genes with high insect resistance; Using gene editing technology, such as CRISPR-Cas9, to introduce the selected insect-resistant genes into the recipient cells of cotton plants; Conduct multi-generation screening and stable genetic testing of cotton plants with introduced insect-resistant genes; b. Screening and cultivation of low phenolic genes, including: Analyze the key genes for the synthesis of cotton phenols; Screening for genotypes or mutants with low phenolic expression characteristics; Through hybridization and backcrossing, low-phenol characteristics are introduced into the target variety, and multiple generations of selection and stable genetic testing are carried out to eventually cultivate insect-resistant, low-phenol high-quality cotton varieties.

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