Breeding method of high-temperature-resistant and high-quality cotton hybrid and intelligent breeding system

Abstract

The invention belongs to the technical field of breeding of cotton hybrids, and discloses a breeding method and an intelligent breeding system of high-temperature-resistant high-quality cotton hybrids, which improve the high-temperature resistance: breeding a cotton variety with remarkably improved high-temperature resistance through a gene screening and introducing technology, and reducing the negative influence of high temperature on cotton growth. An intelligent breeding system and a high-throughput genome sequencing technology are applied, so that the hybridization efficiency is remarkably improved, and the breeding period is shortened. By using the intelligent breeding platform, the automation and intelligence of the breeding process can be realized, the manual operation is reduced, and the breeding cost is reduced. Through multi-environment tests and data analysis, the bred cotton variety has better environmental adaptability and can be popularized and applied in different areas, and the cotton yield and quality are improved.

Description

Technical Field

[0001] The present invention belongs to the technical field of cotton hybrid breeding, and in particular relates to a breeding method and an intelligent breeding system for high-temperature resistant high-quality cotton hybrids. Background Art

[0002] Existing cotton varieties are not resistant to high temperatures: Existing cotton varieties are limited in growth under high temperature conditions, resulting in reduced yield and quality. Low hybridization efficiency: Traditional hybridization methods are time-consuming and labor-intensive, making it difficult to efficiently obtain high-quality hybrids. Long breeding cycle: It takes multiple growth cycles from initial hybridization to the final selection of high-quality cotton varieties, which is time-consuming. Poor environmental adaptability: Existing varieties perform inconsistently in different environments, making them difficult to promote and apply.

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

[0004] (1) Insufficient high temperature resistance: Existing cotton varieties are restricted in growth under high temperature conditions, resulting in reduced yield and quality.

[0005] (2) Low hybridization efficiency: Traditional hybridization methods are time-consuming and labor-intensive, making it difficult to efficiently obtain high-quality hybrids.

[0006] (3) Long breeding cycle: It takes multiple growth cycles from initial hybridization to the final selection of high-quality cotton varieties, which is time-consuming.

[0007] (4) Poor environmental adaptability: Existing varieties perform inconsistently in different environments, making them difficult to promote and apply. Summary of the invention

[0008] In view of the problems existing in the prior art, the present invention provides a breeding method and an intelligent breeding system for high-temperature resistant and high-quality cotton hybrids.

[0009] The present invention is achieved in that a breeding method for high-temperature resistant high-quality cotton hybrids comprises:

[0010] Step 1: Field phenotypic identification in high temperature areas combined with molecular marker-assisted selection;

[0011] Step 2, gene introduction;

[0012] Step 3, high-throughput genome sequencing:

[0013] Genome sequencing: Perform high-throughput genome sequencing on different cotton varieties to obtain complete genome sequence data;

[0014] High-quality gene screening: Use bioinformatics analysis tools to screen out key genes related to high-quality traits including high temperature resistance, high yield, disease resistance, and fiber quality.

[0015] Furthermore, the field phenotypic identification in high temperature areas is combined with a molecular marker-assisted selection method:

[0016] 1) Target gene screening: Using the existing cotton genome database and combining the cotton phenotypic data under high temperature stress, a genome-wide association analysis (GWAS) was performed to screen out genes significantly associated with high temperature tolerance traits;

[0017] 2) Molecular marker development.

[0018] Furthermore, the molecular marker development: developing specific molecular markers for the screened target genes to assist in the selection of high temperature resistance traits.

[0019] Furthermore, the gene introduction method:

[0020] (1) Gene editing technology:

[0021] (2) Genetically modified technology.

[0022] Furthermore, the gene editing technology:

[0023] CRISPR / Cas9 or TALEN gene editing tools are used to perform gene editing on target cotton varieties and precisely introduce the screened high-temperature-resistant genes.

[0024] Furthermore, the transgenic technology:

[0025] Using Agrobacterium-mediated method or gene gun technology, the high temperature resistance gene is transferred into the genome of the target cotton variety to generate transgenic plants.

[0026] Another object of the present invention is to provide a breeding system for high-temperature resistant high-quality cotton hybrids, comprising:

[0027] Data management module, used to manage and store genomic data, phenotypic data, and environmental data, and provide data retrieval and analysis functions;

[0028] Algorithm module, which is used to integrate big data analysis, machine learning and artificial intelligence algorithms to achieve hybrid combination prediction and breeding effect evaluation;

[0029] The user interface module is used to provide a friendly user interface to support breeding experts in data query, program design and result viewing.

[0030] 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 method for breeding high-temperature resistant high-quality cotton hybrids.

[0031] 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 method for breeding high-temperature resistant high-quality cotton hybrids.

[0032] Another object of the present invention is to provide an information data processing terminal, which is used to implement the breeding system of the high-temperature resistant high-quality cotton hybrids.

[0033] 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:

[0034] First, improve high temperature resistance: Through gene screening and introduction technology, select improved cotton varieties with significantly improved high temperature resistance to reduce the negative impact of high temperature on cotton growth.

[0035] Improve breeding efficiency: Apply intelligent breeding systems and high-throughput genome sequencing technology to significantly improve hybridization efficiency and shorten the breeding cycle.

[0036] Optimize the breeding process: The use of intelligent breeding platforms can realize the automation and intelligence of the breeding process, reduce manual operations, and reduce breeding costs.

[0037] Enhance environmental adaptability: Through multi-environment high temperature resistance tests and data analysis, the selected cotton varieties have better environmental adaptability and can be promoted and applied in different regions to improve cotton yield and quality.

[0038] Second, the present invention solves many technical problems in the prior art in the cotton breeding process, such as high temperature resistance, disease resistance, high yield and fiber quality improvement. Traditional 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, through the combination of molecular marker-assisted selection and high-throughput genome sequencing technology, the present invention can quickly and accurately screen out key genes related to high-quality traits, thereby greatly shortening the breeding cycle and improving breeding efficiency.

[0039] In terms of gene introduction, the present invention uses modern genetic engineering technology to overcome the problems of low gene integration efficiency and poor stability in traditional breeding methods. Through advanced gene introduction technologies such as Agrobacterium-mediated method and gene gun method, the present invention achieves efficient integration and stable expression of target genes, thereby significantly improving the heat resistance and disease resistance of cotton varieties. This technological advancement enables cotton to maintain high yield and high-quality fiber characteristics in the face of harsh environmental conditions.

[0040] In addition, the present invention can fully understand the structure and function of the cotton genome through high-throughput genome sequencing and bioinformatics analysis. This in-depth genome analysis capability enables the present invention to accurately locate genes related to high-quality traits, thereby performing targeted gene improvement and optimization. Compared with traditional empirical breeding methods, this genome-level improvement method is not only more scientific and reasonable, but also can significantly improve the overall performance and market competitiveness of cotton varieties.

[0041] In summary, the present invention has achieved significant improvements in cotton varieties in terms of heat resistance, disease resistance, high yield and fiber quality through the comprehensive application of field phenotypic identification in high temperature zones combined with molecular marker-assisted selection, gene introduction and high-throughput genome sequencing. Experimental data show that the cotton varieties (lines) bred by the present invention are superior to existing varieties (lines) in multiple key performance indicators, demonstrating significant technological progress and broad application prospects. This innovative breeding method provides a scientific and effective technical path for cotton breeding, and is expected to promote the further development and upgrading of the cotton industry in the future. BRIEF DESCRIPTION OF THE DRAWINGS

[0042] Figure 1 The present invention provides a flow chart of a breeding method for high-temperature resistant high-quality cotton hybrids.

[0043] Figure 2 It is a flow chart of the molecular marker-assisted selection method provided by an embodiment of the present invention.

[0044] Figure 3 It is a flow chart of the gene introduction method provided by an embodiment of the present invention.

[0045] Figure 4 It is a structural block diagram of a breeding system for high-temperature resistant high-quality cotton hybrids provided by an embodiment of the present invention. DETAILED DESCRIPTION

[0046] 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.

[0047] The following is a specific implementation method for the breeding method of high-temperature resistant high-quality cotton hybrids:

[0048] 1) Molecular marker-assisted selection:

[0049] 1.1 Use high-throughput genome sequencing technology to sequence the genomes of multiple cotton varieties, using Illumina or PacBio platforms as sequencing equipment;

[0050] 1.2 Use bioinformatics analysis tools (such as GATK, BWA, and SAMtools) to compare and detect variations in genome sequences, and screen for molecular markers related to heat resistance, disease resistance, high yield, and high-quality fiber;

[0051] 1.3 Use marker-assisted selection (MAS) technology to detect target molecular markers through quantitative PCR (qPCR) or KASP marker technology to select individuals with target traits;

[0052] 1.4 Through multi-generation screening, individuals showing high temperature resistance, disease resistance, and high-quality fiber traits are retained, hybridization and backcrossing are carried out, and the stable inheritance of molecular markers is maintained in the offspring.

[0053] 2) Gene introduction:

[0054] 2.1 Clone relevant target genes from plants or other organisms that are resistant to high temperatures, diseases, high-yield or high-quality fibers, and use CRISPR-Cas9, TALEN or ZFN gene editing tools to precisely edit the target genes;

[0055] 2.2 Construct the target gene into a plant expression vector (such as pCAMBIA, pBI121), and introduce it into cotton callus cells using Agrobacterium-mediated method or gene gun method;

[0056] 2.3 Induce and regenerate complete transgenic cotton plants through tissue culture technology, and conduct genotype screening to detect gene integration;

[0057] 2.4 Use PCR and Southern blot techniques to verify the integration of the target gene in the cotton genome, and use qPCR to detect the expression level of the target gene to ensure the effectiveness and stability of gene expression.

[0058] 3) Genome sequencing and high-quality gene screening:

[0059] 3.1 Perform high-throughput genome sequencing on cotton plants with the target gene introduced, and obtain whole genome sequence data using the Illumina NovaSeq or Oxford Nanopore platform;

[0060] 3.2 Use bioinformatics analysis tools (such as FASTQC, Bowtie2, DESeq2) to analyze sequencing data, screen out key genes related to traits such as high-quality fiber, high temperature resistance, disease resistance, and high yield, and verify gene functions in combination with RNA-Seq data;

[0061] 3.3 Further verify the function of the selected key genes in cotton through transgenic technology or RNA interference (RNAi) technology, and evaluate their effects on target traits (such as heat resistance, disease resistance, and high-quality fiber);

[0062] 3.4 Use molecular marker-assisted technology to maintain the genetic stability of these high-quality genes in subsequent breeding, and ultimately select cotton hybrids that are heat-resistant, disease-resistant, and have high-quality fibers.

[0063] Further including:

[0064] 1) Collection and analysis of phenotypic data:

[0065] 1.1 Use a high-throughput phenotyping system to collect phenotypic data of cotton during breeding, including cotton's high temperature resistance traits (such as leaf temperature and stomatal conductance), fiber quality (such as length and strength), and disease resistance;

[0066] 1.2 Monitor the growth status of cotton in the field through IoT devices, use multispectral imagers and infrared thermal imagers to detect the temperature and moisture conditions of the cotton growth environment, and conduct comprehensive analysis based on greenhouse experimental data;

[0067] 1.3 Use machine learning algorithms to analyze phenotypic data, predict cotton growth trends and target trait performance, and optimize breeding strategies.

[0068] The method includes using RNA-Seq technology to analyze the transcriptome of transgenic cotton, screening out differently expressed genes (DEGs) related to the target trait, and combining proteomics analysis to further confirm the mechanism of action of the target gene.

[0069] This specific claim details the specific steps of molecular markers, gene introduction, genome sequencing and phenotypic data collection, and combines modern breeding technology and data analysis methods to make the breeding method more practical and scientific.

[0070] The present invention proposes a breeding method for high-temperature resistant high-quality cotton hybrids, which involves three steps: molecular marker-assisted selection, gene introduction, and high-throughput genome sequencing. The following is a detailed working principle:

[0071] Step 1: Marker-assisted selection

[0072] Marker-assisted selection (MAS) is a method that uses known genomic information to select individuals with superior traits. It works as follows:

[0073] 1) Marker development: First, the genomes of different cotton varieties were sequenced using high-throughput genome sequencing technology to obtain complete genome sequence data.

[0074] 2) Marker screening: Use bioinformatics tools to analyze sequencing data and screen out molecular markers related to traits such as high temperature tolerance, high-quality fiber, and disease resistance.

[0075] 3) Marker verification: Experimentally verify the accuracy and stability of these molecular markers in target traits.

[0076] 4) Marker application: In the cotton breeding process, these molecular markers are detected to select individuals with target traits and accelerate the breeding process.

[0077] Step 2: Gene introduction

[0078] Gene introduction refers to the introduction of target genes into the cotton genome through genetic engineering technology to obtain cotton varieties with specific excellent traits. Its working principle is as follows:

[0079] 1) Gene cloning: Clone target genes related to heat resistance, disease resistance, high yield or high-quality fiber from other plants or organisms.

[0080] 2) Vector construction: Insert the cloned target gene into an appropriate plant expression vector to construct a gene introduction vector.

[0081] 3) Gene transformation: Gene transformation techniques such as Agrobacterium-mediated method or gene gun method are used to introduce the target gene into cotton cells.

[0082] 4) Regenerated plants: Through tissue culture technology, the transformed cotton cells are cultured into complete plants, and the individuals with successfully introduced target genes are screened.

[0083] 5) Gene verification: The integration and expression level of the target gene in the cotton genome are verified through molecular biological methods such as PCR and Southern blot.

[0084] Step 3: High-throughput genome sequencing

[0085] High-throughput genome sequencing is a technology that can quickly obtain a large amount of genome sequence information, which is used to fully understand the genome structure and function of cotton varieties. Its working principle is as follows:

[0086] 1) Genome sequencing: Perform high-throughput sequencing on the genomes of different cotton varieties to obtain a large amount of genome sequence data.

[0087] 2) Data analysis: Use bioinformatics analysis tools to process and analyze sequencing data and construct a genome sequence map.

[0088] 3) High-quality gene screening: Through comparative analysis, key genes related to high-quality traits (such as high yield, disease resistance, fiber quality, etc.) are screened out.

[0089] 4) Functional verification: Use transgenic technology or other experimental methods to verify the functions and effects of these key genes in cotton.

[0090] Specific examples:

[0091] 1) Screening of high temperature resistance genes: Through genome sequencing and bioinformatics analysis, genes related to high temperature resistance traits, such as heat shock protein genes (HSPs), are found.

[0092] 2) Disease resistance gene screening: Screen out genes related to disease resistance, such as disease resistance R gene, and enhance cotton's disease resistance through gene introduction technology.

[0093] 3) Fiber quality improvement: Screen out genes related to fiber length, strength and toughness, and conduct molecular marker-assisted selection and gene introduction to improve the fiber quality of cotton.

[0094] In summary, the present invention systematically screens and introduces key genes related to high-quality traits through field phenotypic identification in high-temperature zones combined with molecular marker-assisted selection, gene introduction, and high-throughput genome sequencing, significantly improving the breeding efficiency and quality of cotton varieties. Experimental data show that this method has made significant progress in improving high temperature resistance, disease resistance, high yield, and high-quality fiber traits, providing a scientific and effective technical means for cotton breeding.

[0095] like Figure 1 As shown, a breeding method for high-temperature resistant high-quality cotton hybrids provided by an embodiment of the present invention comprises the following steps:

[0096] S101, field phenotyping combined with molecular marker-assisted selection in high temperature areas;

[0097] S102, gene introduction;

[0098] S103, high-throughput genome sequencing:

[0099] Genome sequencing: Perform high-throughput genome sequencing on different cotton varieties to obtain complete genome sequence data;

[0100] High-quality gene screening: Use bioinformatics analysis tools to screen out key genes related to high-quality traits including high yield, disease resistance, and fiber quality.

[0101] like Figure 2 As shown, the molecular marker-assisted selection method provided by the embodiment of the present invention is:

[0102] S201, target gene screening: using the existing cotton genome database, combined with cotton phenotypic data under high temperature stress, to conduct genome-wide association analysis (GWAS) to screen out genes significantly associated with high temperature tolerance traits;

[0103] S202, Molecular marker development.

[0104] The molecular marker development provided by the embodiment of the present invention is: for the screened target gene, a specific molecular marker is developed to assist in the selection of high temperature resistance traits.

[0105] like Figure 3 As shown, the gene introduction method provided by the embodiment of the present invention is:

[0106] S301, gene editing technology:

[0107] S302, genetic modification technology.

[0108] The gene editing technology provided by the embodiments of the present invention:

[0109] CRISPR / Cas9 or TALEN gene editing tools are used to perform gene editing on target cotton varieties and precisely introduce the screened high-temperature-resistant genes.

[0110] The transgenic technology provided by the embodiments of the present invention:

[0111] Using Agrobacterium-mediated method or gene gun technology, the high temperature resistance gene is transferred into the genome of the target cotton variety to generate transgenic plants.

[0112] like Figure 4 As shown, a breeding system for high-temperature resistant high-quality cotton hybrids provided by an embodiment of the present invention comprises:

[0113] Data management module, used to manage and store genomic data, phenotypic data, and environmental data, and provide data retrieval and analysis functions;

[0114] Algorithm module, which is used to integrate big data analysis, machine learning and artificial intelligence algorithms to achieve hybrid combination prediction and breeding effect evaluation;

[0115] The user interface module is used to provide a friendly user interface to support breeding experts in data query, program design and result viewing.

[0116] 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 method for breeding high-temperature resistant high-quality cotton hybrids.

[0117] 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 method for breeding high-temperature resistant high-quality cotton hybrids.

[0118] Another object of the present invention is to provide an information data processing terminal, which is used to implement the breeding system of the high-temperature resistant high-quality cotton hybrids.

[0119] The present invention is specifically implemented:

[0120] 1. Screening and introduction of high temperature tolerance genes

[0121] 1. Molecular marker-assisted selection technology:

[0122] Target gene screening: Using the existing cotton genome database and combining it with cotton phenotypic data under high temperature stress, a genome-wide association study (GWAS) was performed to screen out genes that are significantly associated with high temperature tolerance traits.

[0123] Molecular marker development: Develop specific molecular markers for the screened target genes to assist in the selection of high temperature resistance traits.

[0124] 2. Gene introduction technology:

[0125] Gene editing technology: Use gene editing tools such as CRISPR / Cas9 or TALEN to edit the genes of target cotton varieties and accurately introduce the screened high-temperature resistant genes.

[0126] Transgenic technology: Use Agrobacterium-mediated or gene gun technology to transfer high temperature resistance genes into the genome of the target cotton variety to generate transgenic plants.

[0127] 2. Efficient breeding methods for high-quality cotton hybrids

[0128] 1. High-throughput genome sequencing technology:

[0129] Genome sequencing: High-throughput genome sequencing is performed on different cotton varieties to obtain complete genome sequence data.

[0130] High-quality gene screening: Use bioinformatics analysis tools to screen out key genes related to high-quality traits (such as high yield, disease resistance, fiber quality, etc.).

[0131] 2. Intelligent breeding system:

[0132] Data integration and management: Build a cotton genome database, integrate genome data, phenotypic data and environmental data, and provide data support for intelligent breeding.

[0133] Hybrid combination prediction: Apply big data analysis and artificial intelligence algorithms to predict the optimal hybrid combinations among different cotton varieties and improve the success rate and effect of hybridization.

[0134] Breeding plan design: Based on the prediction results, specific breeding plans are automatically generated, including hybridization strategies, cultivation management, etc.

[0135] 3. Construction of Intelligent Breeding System

[0136] 1. System architecture design:

[0137] Data management module: manages and stores genomic data, phenotypic data, and environmental data, and provides data retrieval and analysis functions.

[0138] Algorithm module: Integrates big data analysis, machine learning and artificial intelligence algorithms to achieve hybrid combination prediction and breeding effect evaluation.

[0139] User interface module: provides a friendly user interface to support breeding experts in data query, program design and result viewing.

[0140] 2. Technology integration and optimization:

[0141] Big data analysis platform: Uses big data processing frameworks such as Hadoop and Spark to achieve efficient processing and analysis of massive genomic data.

[0142] Artificial intelligence algorithms: Apply machine learning algorithms such as deep learning and random forests to improve the accuracy and reliability of breeding programs.

[0143] 4. Environmental adaptability test

[0144] 1. Multi-environment field trials:

[0145] Experimental design: Design reasonable field experimental plans to plant and manage hybrids under different ecological environments (such as high temperature, drought, saline-alkali land, etc.).

[0146] Data collection: Use sensors, drones and remote sensing technology to monitor cotton growth in real time and collect multi-dimensional phenotypic data.

[0147] 2. Data analysis and optimization:

[0148] Data analysis: Apply big data analysis tools to conduct comprehensive analysis of multi-environment test data and evaluate the performance of different hybrids.

[0149] Strategy optimization: Based on the test results, optimize the breeding strategy and select high-quality cotton varieties that are adapted to different environmental conditions.

[0150] V. Specific implementation steps

[0151] 1. Gene screening and introduction:

[0152] Collect and analyze genomic and phenotypic data of cotton varieties, conduct GWAS analysis, and screen for heat resistance genes.

[0153] Using gene editing and transgenic technology, the screened high-temperature-resistant genes are introduced into the target cotton varieties to generate transgenic plants.

[0154] 2. High-quality gene screening and hybridization:

[0155] Perform high-throughput genome sequencing on different cotton varieties to screen high-quality genes.

[0156] Through the intelligent breeding system, the optimal hybrid combination is designed and hybridization experiments are carried out.

[0157] 3. Development of intelligent breeding system:

[0158] Build data management, algorithm and user interface modules, integrate big data and artificial intelligence technologies, and develop an intelligent breeding platform.

[0159] Conduct system testing and optimization to ensure the stability and reliability of system functions.

[0160] 4. Environmental adaptability test:

[0161] Conduct field experiments in different ecological environments and collect multi-dimensional data.

[0162] Analyze experimental data, optimize breeding strategies, and select high-quality cotton varieties that are adapted to different environments.

[0163] Through the above-mentioned specific implementation plan, the technical problems faced in the cotton breeding process can be effectively solved, the breeding efficiency and the high temperature resistance of cotton varieties can be significantly improved, and the rapid selection and promotion of high-quality cotton varieties can be achieved.

[0164] 3. Technical Effects

[0165] 1. Improve high temperature resistance: Through gene screening and introduction technology, cotton varieties with significantly improved high temperature resistance are bred to reduce the negative impact of high temperature on cotton growth.

[0166] 2. Improve breeding efficiency: Apply intelligent breeding systems and high-throughput genome sequencing technology to significantly improve hybridization efficiency and shorten the breeding cycle.

[0167] 3. Optimize the breeding process: The use of intelligent breeding platforms can realize the automation and intelligence of the breeding process, reduce manual operations, and reduce breeding costs.

[0168] 4. Enhance environmental adaptability: Through multi-environment tests and data analysis, the selected cotton varieties have better environmental adaptability and can be promoted and applied in different regions to improve cotton yield and quality.

[0169] IV. Implementation steps

[0170] 1. Screening of genes for high temperature tolerance:

[0171] Collect genomic data from different cotton varieties.

[0172] Genes related to high temperature tolerance were screened out through genome-wide association study (GWAS).

[0173] 2. Gene introduction and hybridization:

[0174] Using gene editing technology, high temperature resistance genes are introduced into target cotton varieties.

[0175] Design the optimal hybrid combination through the intelligent breeding system and conduct hybridization experiments.

[0176] 3. Development and application of intelligent breeding system:

[0177] Develop intelligent breeding systems, including genome data management modules, hybrid combination prediction modules, breeding effect evaluation modules, etc.

[0178] The screened genomic data is input into the system to automatically design and implement breeding plans.

[0179] 4. Field trials and optimization:

[0180] Field trials were conducted in different ecological environments to collect cotton growth data.

[0181] Based on the test results, the breeding plan is adjusted and optimized to finally select high-quality cotton varieties.

[0182] Through the above technical solution, the problems existing in the current cotton breeding process, such as insufficient high temperature resistance, low hybridization efficiency, long breeding cycle and poor environmental adaptability, can be effectively solved, and the high temperature resistance and overall quality of cotton varieties can be significantly improved.

[0183] The method for breeding high-quality high-temperature resistant cotton hybrids of the present invention has achieved remarkable positive results 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:

[0184] First, the present invention significantly improves breeding efficiency through molecular marker-assisted selection technology. In the experiment, multiple cotton varieties were sequenced using high-throughput genome sequencing technology to screen out molecular markers related to high temperature resistance, high yield, disease resistance and high-quality fiber. Combining these markers for selection shortened the breeding cycle by about 30% and significantly improved the accuracy and efficiency of breeding.

[0185] Secondly, in the process of gene introduction, the present invention uses Agrobacterium-mediated method and gene gun method to successfully clone and introduce target genes related to excellent traits. In multiple experiments, the success rate of gene introduction reached more than 85%, which is much higher than the 50% of traditional methods. The introduced genes show good stability and expression level in cotton cells, which significantly improves the heat resistance and disease resistance of the newly cultivated cotton varieties.

[0186] In addition, through high-throughput genome sequencing and bioinformatics analysis, the present invention accurately screened out a number of key genes related to high-quality traits. In the experiment, the cotton varieties improved by using these key genes increased their yield by 20% and their fiber quality (such as length and strength) by 15%. These data show that the breeding method of the present invention is not only scientific in theory, but also has achieved remarkable results in practical applications.

[0187] Finally, through comprehensive test data analysis, the method for breeding high-temperature resistant high-quality cotton hybrids of the present invention is superior to existing varieties in multiple key performance indicators. The following table shows the comparative data of the cotton varieties bred by the present invention and traditional varieties in terms of high temperature resistance, disease resistance, high yield and fiber quality:

[0188]

[0189] It can be seen from the above data that the present invention has achieved remarkable technical progress in terms of high temperature resistance, disease resistance, high yield and fiber quality, demonstrating its great potential and broad application prospects in cotton breeding.

[0190] The following are two specific embodiments, based on the practical application of the above method:

[0191] Example 1: Breeding of high temperature resistant cotton varieties

[0192] 1. Marker-assisted selection (MAS): Through high-throughput sequencing of the genomes of multiple heat-resistant cotton varieties, molecular markers closely related to heat-resistant traits were screened out. These markers are located in specific regions of the cotton genome, and their association with heat-resistant genes is confirmed through bioinformatics analysis.

[0193] 2. Gene introduction: Known heat-resistant related genes (such as the HSPs gene family) are cloned from heat-resistant plants such as rice or Arabidopsis, and introduced into cotton cells using the Agrobacterium-mediated method. After gene transformation and tissue culture, cotton plants with heat-resistant characteristics are obtained.

[0194] 3. Verification and screening: Through molecular marker detection, individuals with successfully introduced high temperature resistance genes were screened out, and field high temperature stress tests were conducted. The results showed that the newly bred cotton varieties had a higher survival rate and better growth performance under high temperature environments.

[0195] Example 2: Breeding of high-quality fiber cotton varieties

[0196] 1. Marker-assisted selection (MAS): Using high-throughput genome sequencing technology, molecular markers related to high-quality fiber traits (such as long-staple fiber and twist resistance) are screened for multiple cotton varieties. These markers are related to cotton fiber strength, length, and toughness.

[0197] 2. Gene introduction: Cloning genes related to fiber quality improvement (such as GhMYB25-like and GhHD-1) and introducing them into cotton cells through gene gun technology. Subsequently, transgenic plants are cultivated through tissue culture methods.

[0198] 3. Verification and screening: PCR testing confirmed that the fiber quality-related genes were successfully integrated into the cotton genome. Field experiments were conducted to measure fiber length, strength and anti-twisting properties. The results showed that the fiber quality of this cotton variety was significantly better than that of traditional varieties and had a high application value in the market.

[0199] These two examples demonstrate how cotton breeding technology for heat resistance and high-quality fiber traits can apply gene screening and introduction technology to accelerate the selection of new varieties.

[0200] 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 modifications, equivalent substitutions and improvements 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 high-temperature resistant high-quality cotton hybrids, characterized in that it comprises the following steps: 1) Molecular marker-assisted selection: 1.1 Use high-throughput genome sequencing technology to sequence the genomes of multiple cotton varieties, using Illumina or PacBio platforms as sequencing equipment; 1.2 Use bioinformatics analysis tools (such as GATK, BWA, and SAMtools) to compare and detect variations in genome sequences, and screen for molecular markers related to heat resistance, disease resistance, high yield, and high-quality fiber; 1.3 Use marker-assisted selection (MAS) technology to detect target molecular markers through quantitative PCR (qPCR) or KASP marker technology to select individuals with target traits; 1.4 Through multi-generation screening, individuals showing high temperature resistance, disease resistance, and high-quality fiber traits are retained, hybridization and backcrossing are carried out, and the stable inheritance of molecular markers is maintained in the offspring. 2) Gene introduction: 2.1 Clone relevant target genes from plants or other organisms that are resistant to high temperatures, diseases, high-yield or high-quality fibers, and use CRISPR-Cas9, TALEN or ZFN gene editing tools to precisely edit the target genes; 2.2 Construct the target gene into a plant expression vector (such as pCAMBIA, pBI121), and introduce it into cotton callus cells using Agrobacterium-mediated method or gene gun method; 2.3 Induce and regenerate complete transgenic cotton plants through tissue culture technology, and conduct genotype screening to detect gene integration; 2.4 Use PCR and Southern blot techniques to verify the integration of the target gene in the cotton genome, and use qPCR to detect the expression level of the target gene to ensure the effectiveness and stability of gene expression. 3) Genome sequencing and high-quality gene screening: 3.1 Perform high-throughput genome sequencing on cotton plants with the target gene introduced, and obtain whole genome sequence data using the Illumina NovaSeq or Oxford Nanopore platform; 3.2 Use bioinformatics analysis tools (such as FASTQC, Bowtie2, DESeq2) to analyze sequencing data, screen out key genes related to traits such as high-quality fiber, high temperature resistance, disease resistance, and high yield, and verify gene functions in combination with RNA-Seq data; 3.3 Further verify the function of the selected key genes in cotton through transgenic technology or RNA interference (RNAi) technology, and evaluate their effects on target traits (such as heat resistance, disease resistance, and high-quality fiber); 3.4 Use molecular marker-assisted technology to maintain the genetic stability of these high-quality genes in subsequent breeding, and ultimately select cotton hybrids that are heat-resistant, disease-resistant, and have high-quality fibers.

2. The breeding method of heat-resistant high-quality cotton hybrids according to claim 1, further comprising: 1) Collection and analysis of phenotypic data: 1.1 Use a high-throughput phenotyping system to collect phenotypic data of cotton during breeding, including cotton's high temperature resistance traits (such as leaf temperature and stomatal conductance), fiber quality (such as length and strength), and disease resistance; 1.2 Monitor the growth status of cotton in the field through IoT devices, use multispectral imagers and infrared thermal imagers to detect the temperature and moisture conditions of the cotton growth environment, and conduct comprehensive analysis based on greenhouse experimental data; 1.3 Use machine learning algorithms to analyze phenotypic data, predict cotton growth trends and target trait performance, and optimize breeding strategies.

3. The breeding method of high temperature resistant high quality cotton hybrid according to claim 1, It is characterized in that The method includes using RNA-Seq technology to analyze the transcriptome of transgenic cotton, screening out differently expressed genes (DEGs) related to target traits, and combining proteomics analysis to further confirm the mechanism of action of the target gene.

4. The method for breeding a high temperature resistant high quality cotton hybrid according to claim 1, It is characterized in that The following steps are involved: Molecular marker-assisted selection: Use high-throughput genome sequencing technology to sequence the genomes of different cotton varieties and obtain complete genome sequence data; use bioinformatics analysis tools to screen out molecular markers related to high temperature resistance, high yield, disease resistance and high-quality fiber, and verify the accuracy and stability of these molecular markers through experiments; in the breeding process, select individuals with target traits by detecting these molecular markers; Gene introduction: clone target genes related to heat resistance, disease resistance, high yield or high-quality fiber from other plants or organisms, and insert these genes into plant expression vectors; introduce target genes into cotton cells through Agrobacterium-mediated method or gene gun method, and cultivate them into complete plants using tissue culture technology; verify the integration and expression of target genes in the cotton genome through PCR and Southern blot methods; Genome sequencing and high-quality gene screening: Perform high-throughput genome sequencing on cotton varieties with introduced target genes to obtain genome sequence data; use bioinformatics tools to analyze data, screen out key genes related to high-quality traits, and verify the functions and effects of these key genes in cotton through transgenic technology or other experimental means. The method is characterized in that the molecular marker-assisted selection step further comprises: Marker development: Development of new molecular markers based on genome sequencing data; Marker application: In the cotton breeding process, individuals with target traits can be quickly screened out through high-throughput molecular marker detection technology; Target gene screening: Through gene introduction and selection technology, cotton plants with high temperature resistance, high yield, disease resistance or high-quality fiber traits are screened out; Data integration and analysis: Integrate molecular marker data and genome sequencing data, and use bioinformatics analysis tools to conduct large-scale data analysis and high-quality gene screening to guide the improvement and optimization of cotton varieties.

5. The breeding method of high temperature resistant high quality cotton hybrid according to claim 1, It is characterized in that The molecular marker assisted selection method: 1) Target gene screening: Using the existing cotton genome database and combining the cotton phenotypic data under high temperature stress, a genome-wide association analysis (GWAS) was performed to screen out genes significantly associated with high temperature tolerance traits; 2) Molecular marker development.

6. The breeding method of high temperature resistant high quality cotton hybrid according to claim 1, It is characterized in that The molecular marker development: develops specific molecular markers for the screened target genes to assist in the selection of high temperature resistance traits; the gene editing technology uses CRISPR / Cas9 or TALEN gene editing tools to perform gene editing on the target cotton variety and accurately introduce the screened high temperature resistance genes.

7. The breeding method of high temperature resistant high quality cotton hybrids as claimed in claim 6, It is characterized in that The transgenic technology: Using Agrobacterium-mediated method or gene gun technology, the high temperature resistance gene is transferred into the genome of the target cotton variety to generate transgenic plants.

8. A breeding system for heat-resistant high-quality cotton hybrids for implementing the breeding method for heat-resistant high-quality cotton hybrids according to any one of claims 1 to 7, It is characterized in that The breeding system of the high temperature resistant high quality cotton hybrids comprises: Data management module, used to manage and store genomic data, phenotypic data, and environmental data, and provide data retrieval and analysis functions; Algorithm module, which is used to integrate big data analysis, machine learning and artificial intelligence algorithms to achieve hybrid combination prediction and breeding effect evaluation; The user interface module is used to provide a friendly user interface to support breeding experts in data query, program design and result viewing.

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