Construction method and application of molecular identity card of germplasm resource library
By using a variety of molecular marker technologies in the germplasm resource library to construct molecular ID cards, the accuracy and uniqueness of germplasm resource management in traditional methods are solved, and efficient and accurate germplasm resource management and utilization are achieved.
Patent Information
- Application Number
- CN202510325156.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-03-19
- Publication Date
- 2025-06-20
AI Technical Summary
It is difficult for the existing technology to achieve accurate and unique germplasm resource management in large-scale germplasm resource databases. Traditional morphological characteristic methods have problems such as low accuracy, easy to confuse and lack of uniqueness.
A variety of molecular marker technologies (such as SSR and SNP) are used to construct a unique molecular ID card for each germplasm resource through DNA extraction, PCR amplification and detection, and integrate and represent it in the form of digital encoding, barcode or QR code, and establish a database.
It significantly improves the accuracy and uniqueness of germplasm resource identification, optimizes the management efficiency of germplasm resource library, improves the utilization efficiency and protection capabilities of germplasm resources, and promotes biodiversity research.
Smart Images
Figure CN120174136A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the technical field of germplasm resource protection and management, and more specifically, it relates to a method for constructing a molecular identity card for a germplasm resource bank and its application. Background Art
[0002] Germplasm resources are an important part of biodiversity and are of great significance for the sustainable development of fields such as agriculture, forestry, and horticulture. A germplasm resource bank is an important facility for preserving and managing germplasm resources. However, traditional germplasm resource management mainly relies on morphological characteristics and geographical information, etc. For example, the morphological characteristics of plants (such as leaf shape, flower color, fruit size, etc.) and the appearance characteristics of animals (such as hair color, body shape, etc.) are widely used for the classification and identification of germplasm resources. However, these methods have many limitations: For example, low accuracy: Morphological characteristics are easily affected by environmental factors. The same species may show different morphological characteristics in different growth environments, resulting in inaccurate identification results. For example, the leaf shape and size of plants may vary significantly under different environmental conditions such as light and water. Prone to confusion: Many germplasm resources are extremely similar morphologically and are difficult to distinguish by the naked eye or simple morphological observation. For example, some closely related plant varieties or animal varieties are almost indistinguishable in appearance and are prone to misjudgment. Lack of uniqueness: The classification method based on morphological characteristics cannot provide a unique identity for each germplasm resource and is difficult to meet the needs of refined management of the germplasm resource bank. It mainly relies on morphological characteristics and geographical information, etc., and these methods have problems such as low accuracy and being prone to confusion. With the development of molecular biology technology, using molecular marker technology to construct molecular identity cards for germplasm resources has become a new trend. Molecular identity cards can provide more accurate and unique identity information, which is helpful for the identification, protection, and utilization of germplasm resources. However, there is currently a lack of a systematic, efficient, and applicable method for constructing molecular identity cards for large-scale germplasm resource banks. With the development of molecular biology technology, using molecular marker technology to construct molecular identity cards for germplasm resources has become a new trend. Molecular identity cards can provide more accurate and unique identity information, which is helpful for the identification, protection, and utilization of germplasm resources. However, there is currently a lack of a systematic, efficient, and applicable method for constructing molecular identity cards for large-scale germplasm resource banks. Summary of the Invention
[0003] Aiming at the deficiencies of the existing technology, the purpose of the present invention is to provide a systematic, efficient, and applicable method for constructing molecular identity cards for large-scale germplasm resource banks.
[0004] To achieve the above purpose, the present invention provides the following technical solutions: A method for constructing a molecular identity card for a germplasm resource bank and its application, including the following steps:
[0005] Collect samples of the target germplasm resources from the germplasm resource bank. The sample types include plant leaves, seeds and rhizomes or animal tissues and blood, and number and record the collected samples;
[0006] Extract DNA from the samples using a DNA extraction kit or a self-optimized extraction method, and detect the quality and concentration of DNA by methods such as agarose gel electrophoresis and / or NanoDrop spectrophotometer to ensure the purity and integrity of DNA;
[0007] According to the type of the target germplasm resources, select appropriate molecular marker types, including but not limited to single nucleotide polymorphism (SNP), simple sequence repeat (SSR), amplified fragment length polymorphism (AFLP), etc., design or select specific primers, and use polymerase chain reaction (PCR) technology to amplify the target DNA fragment, and optimize the PCR reaction conditions, including annealing temperature, number of cycles, etc., to ensure the specificity and stability of the amplification product;
[0008] Detect the amplification product, and methods such as gel electrophoresis, capillary electrophoresis, high-throughput sequencing, etc. can be used to analyze the detection results, determine the molecular marker characteristics of each sample, including the type and number of alleles, etc., and organize the analysis results into a data table to record the molecular marker information of each sample;
[0009] According to the combined characteristics of multiple molecular markers, construct a unique molecular identification card for each germplasm resource sample. The molecular identification card can be in the form of digital coding, bar code or two-dimensional code, etc., integrate and represent the molecular marker information, and associate the molecular identification card with other information of the germplasm resources (such as source, morphological characteristics, etc.) to establish a database;
[0010] Verify the constructed molecular identification card. By comparing the molecular marker information of known samples, ensure the accuracy and uniqueness of the molecular identification card, and optimize the selection of molecular markers and amplification conditions according to the verification results to improve the reliability of the molecular identification card.
[0011] In the sample collection step, for plant germplasm resources, select tissue parts with genetic stability and representativeness for sampling, such as leaves, seeds and rhizomes; for animal germplasm resources, select blood and tissue samples to ensure that the quality and content of DNA in the samples meet the requirements of subsequent analysis.
[0012] Preferably, in the DNA extraction step, adopt corresponding extraction methods for different types of samples to improve the DNA extraction efficiency and quality. For example, for plant leaf samples, adopt the improved CTAB method to extract DNA; for animal blood samples, adopt the phenol-chloroform method to extract DNA.
[0013] Preferably, in the step of selecting molecular markers, for plant germplasm resources, SSR markers are preferably selected because of their advantages such as high polymorphism, wide distribution, and easy detection; for animal germplasm resources, SNP markers are preferably selected because of their characteristics such as large quantity, wide distribution, and good genetic stability.
[0014] Preferably, in the step of detecting molecular markers, for SSR markers, capillary electrophoresis is used for detection, which can accurately measure the size and quantity of alleles; for SNP markers, high-throughput sequencing technology is used for detection, which can quickly and accurately obtain genotype information of a large number of SNP loci.
[0015] Preferably, in the step of constructing the molecular identity card, the molecular identity card is represented in the form of a two-dimensional code, which can efficiently integrate and store multiple molecular marker information, and is also convenient for quick reading and query through scanning devices.
[0016] Preferably, in the step of verification and optimization, by comparing the molecular marker information of known samples, the matching degree between the constructed molecular identity card and the known samples is calculated. When the matching degree is lower than the preset threshold, the selection of molecular markers and amplification conditions are optimized to improve the accuracy and uniqueness of the molecular identity card.
[0017] Application of the method for constructing the molecular identity card of the germplasm resource library according to any one of the above in at least one of the following aspects:
[0018] Germplasm resource identification and classification: Using the molecular identity card to quickly and accurately identify germplasm resources, distinguish different species, varieties or ecotypes, and provide a molecular basis for the classification and phylogenetic research of germplasm resources;
[0019] Germplasm resource protection and management: Tracking the origin and flow of germplasm resources through the molecular identity card, preventing the misuse and loss of germplasm resources, providing a scientific basis for the management and maintenance of the germplasm resource library, and optimizing the preservation conditions and strategies of germplasm resources;
[0020] Germplasm resource utilization and development: In the breeding process, using the molecular identity card to screen germplasm resources with excellent traits, improving the breeding efficiency, and providing technical support for the protection of new plant varieties, intellectual property protection, etc.;
[0021] Biodiversity research: Providing molecular marker data for biodiversity research, analyzing the genetic diversity and genetic relationship of germplasm resources, and providing a scientific basis for the formulation of biodiversity protection strategies.
[0022] The application of the molecular identity cards of the above germplasm resource bank can quickly identify the species, variety or ecotype attribution of unknown germplasm resources by comparing the information in the molecular identity card database in the application of germplasm resource identification and classification, and the identification accuracy is higher than that of traditional morphological identification methods.
[0023] The application of the molecular identity cards of the above germplasm resource bank can, in the application of germplasm resource protection and management, use the germplasm resource traceability system established by the molecular identity cards to monitor the usage of germplasm resources in real time, and promptly discover and prevent the illegal transfer or abuse of germplasm resources.
[0024] The application of the molecular identity cards of the above germplasm resource bank can, in the application of germplasm resource utilization and development, screen out germplasm resources with specific excellent trait genes through the molecular identity cards, which can significantly shorten the breeding cycle and improve the breeding success rate.
[0025] The application of the molecular identity cards of the above germplasm resource bank can, in the application of biodiversity research, construct a more accurate species evolutionary tree based on the molecular identity card data of a large number of germplasm resources, providing more scientific decision-making support for biodiversity protection.
[0026] By adopting the above technical solution, the invention has the following effects:
[0027] Significantly improve the accuracy and uniqueness of germplasm resource identification. Traditional germplasm resource identification mainly relies on morphological characteristics, which is easily affected by environmental factors and is difficult to distinguish germplasm resources with similar morphologies. By combining multiple molecular marker techniques (such as SSR and SNP), the present invention can provide accurate and unique identity identifiers for germplasm resources, avoiding misjudgments in traditional methods. For example, in the management of maize germplasm resources, through the combined analysis of SSR and SNP, germplasm resources containing near-isogenic lines can be accurately classified, and different germplasm resources with similar genetic backgrounds can be distinguished.
[0028] Optimize the management efficiency of the germplasm resource bank. The construction of molecular identity cards enables the germplasm resource bank to conduct more refined classification and management. On the basis of preserving and investigating the basic information and phenotypic information of germplasm resources, the molecular information of germplasm resources is added, helping managers quickly and accurately identify and distinguish different germplasm resources. For example, through molecular identity cards, the problem of unclear attribution of germplasm resources stored in the early stage can be effectively solved, avoiding duplicate storage and name confusion.
[0029] Improve the utilization efficiency of germplasm resources. In the breeding process, molecular identity cards can be used to quickly screen out germplasm resources with excellent trait genes, significantly shortening the breeding cycle and improving the breeding success rate. For example, through molecular identity cards, new germplasm can be traced back to the original variety or other authorized varieties, providing support for the confirmation of new plant varieties.
[0030] Enhance the protection ability of germplasm resources. Molecular identification cards can provide a scientific basis for the protection of germplasm resources. By accurately identifying the identity of germplasm resources, it is possible to better trace their sources and flows, preventing the misuse and loss of germplasm resources. In addition, molecular identification cards can also provide technical support for the management and maintenance of germplasm resource banks, optimizing the preservation conditions and strategies of germplasm resources.
[0031] Promote biodiversity research. Molecular identification cards provide rich data support for biodiversity research. By analyzing the molecular marker information of germplasm resources, it is possible to more accurately evaluate the genetic diversity and genetic relationships of germplasm resources. For example, using the data of molecular identification cards, a more accurate species evolutionary tree can be constructed, providing a scientific basis for the formulation of biodiversity protection strategies.
[0032] Drive the modernization and sustainable development of the seed industry. The application of molecular identification cards helps to promote the accurate identification of germplasm resources, laying a solid foundation for the revitalization of the seed industry. Through the national unified public platform of DNA fingerprint databases for crop varieties, the sharing of germplasm resource information can be realized, providing important technical support for the modernization of the seed industry.
[0033] Support applications and collaborations in multiple fields. Molecular identification cards not only have important application value in the agricultural field, but also can promote the application of germplasm resources in other fields, such as environmental protection, ecological restoration, food processing, bioenergy, etc. For example, genetic resources in germplasm resource banks can be used to develop new food raw materials and additives, improving the nutritional value and taste of food.
[0034] In summary, the method for constructing and applying the molecular identification card of the germplasm resource bank of the present invention can not only significantly improve the scientificity and accuracy of germplasm resource management, but also provide strong support for the protection, utilization and biodiversity research of germplasm resources, having important scientific significance and broad application prospects. Brief Description of the Drawings
[0035] Figure 1 It is a schematic flow chart of the present invention. Detailed Embodiments
[0036] Refer to Figure 1 to Figure 1 To further illustrate an embodiment of the method for constructing and applying the molecular identification card of a germplasm resource bank of the present invention.
[0037] Example 1: Construction and Application of Molecular Identification Cards for Maize Germplasm Resource Banks
[0038] 1. Collection of germplasm resource samples: Select 2,918 representative maize germplasm resource samples from the Maize Germplasm Resource Bank of Jilin Province. When collecting samples, ensure the diversity and representativeness of the samples, covering maize germplasm resources from different sources and varieties. Conduct detailed numbering and recording for each sample, including information such as variety name, place of origin, collection time, etc., for subsequent traceability and management.
[0039] 2. DNA extraction and purification: Extract DNA from maize leaf samples using the modified CTAB method. The specific steps are as follows:
[0040] Cut fresh maize leaves into small pieces, put them into a mortar, and add liquid nitrogen to grind them into powder.
[0041] Transfer the powder to a centrifuge tube according to the ratio of adding 1 mL of CTAB extraction buffer per 100 mg of leaves.
[0042] Incubate in a water bath at 65 °C for 30 minutes, and gently invert the centrifuge tube every 10 minutes during this period.
[0043] Centrifuge at 12,000 rpm for 10 minutes, take the supernatant, add an equal volume of chloroform - isoamyl alcohol (24:1) mixture, and gently invert and mix well.
[0044] Centrifuge at 12,000 rpm for 10 minutes, take the supernatant, add 2 volumes of cold absolute ethanol, and precipitate at -20 °C for 30 minutes.
[0045] Centrifuge at 12,000 rpm for 10 minutes, discard the supernatant, wash the precipitate with 70% ethanol, dry it, and dissolve it in an appropriate amount of TE buffer.
[0046] Detect the quality and concentration of DNA by agarose gel electrophoresis to ensure clear DNA bands and no obvious degradation. If the DNA quality is poor, the extraction steps can be repeated or the extraction conditions can be optimized.
[0047] 3. Molecular marker selection and amplification
[0048] (1) SSR molecular markers: Select 40 pairs of SSR marker primers, which have high polymorphism and stability in maize variety identification. Prepare the reaction system according to the "Technical Regulations for Maize Variety Identification - SSR Marker Method" (NY / T 1432—2014): Reaction system: In a 20 μL system, it contains 10 μL of 2×Taq PCR MasterMix, 1 μL of upstream and downstream primers at 10 μmol / L, 1 μL of template DNA (50 ng / μL), and 8 μL of ddH2O.
[0049] Reaction procedure: pre-denaturation at 94°C for 5 minutes; denaturation at 94°C for 40 seconds, annealing at 60°C for 35 seconds, extension at 72°C for 45 seconds, for a total of 35 cycles; extension at 72°C for 10 minutes, and storage at 4°C.
[0050] The amplified products were mixed in 4 groups with equal volumes and detected by capillary electrophoresis using an AB3730XL DNA analyzer (Applied Biosystems, Waltham, USA). The raw data were collected using Date Collection Ver.1.0 software, and fingerprint analysis was performed using SSR Analyser software to filter out high and low peaks caused by asymmetric primer amplification to ensure consistency and accuracy of data collection.
[0051] (2) SNP molecular markers. 61,214 SNP markers were selected. These markers cover the entire corn genome and can provide high-density genetic information. According to the standard chip process, the sample DNA was amplified, fragmented, precipitated, resuspended, hybridized and washed on the chip: after the gDNA was resuspended, the hybridization solution was added and quality inspection was performed. After the quality inspection was qualified, the chip was hybridized, washed and scanned using GeneTitan MC. The raw data was imported into the Axiom Analysis Suite software, the sample set and analysis parameters were selected, the raw data were clustered and genotyped, and the quality control results, sample analysis results and genotyping data were exported.
[0052] 4. Molecular marker detection and analysis
[0053] (1) SSR marker detection: detect the amplified product through capillary electrophoresis and analyze the size and number of alleles. Import the fingerprint data into the plant variety DNA fingerprint management system to generate the germplasm resource SSR-DNA fingerprint data and map. Convert the fingerprint data into the format of "3-digit code + 1-digit letter code", where the 3-digit code represents the length of the main band fragment of the SSR-DNA fingerprint map, and 0 is used to fill in when it is less than three digits; the 1-digit letter code Y / N represents whether the sample presents a homozygous or heterozygous genotype at the marker site.
[0054] (2) SNP marker detection: GeneTitan MC was used for chip hybridization, washing, and scanning, and the raw data was imported into Axiom Analysis Suite software for genotyping. The fingerprint information of 61,214 SNP sites was converted into a visual barcode to form a SNP molecular ID card.
[0055] 5. Molecular ID Card Construction
[0056] (1) SSR molecular identity cards: Based on the fingerprint information of 40 SSR loci, the data is organized into a 160-bit digital table, and then the table data is converted into a QR code form of SSR molecular identity cards. For example, scanning the SSR molecular identity cards of representative germplasm resources Ji 754 and the local variety Zheng Bai 1 will obtain 160-bit digital information, which intuitively reflects the SSR molecular information of different germplasm resource samples.
[0057] (2) SNP molecular identity cards: The fingerprint information of 61,214 SNP loci is converted into a visual bar code to form SNP molecular identity cards. Through SNP molecular identity cards, the traceability of the whole genome fragments can be realized, which is beneficial to the identification of derivative varieties.
[0058] 6. Verification and optimization
[0059] (1) Verification: By comparing the molecular marker information of known samples, verify the accuracy and uniqueness of the molecular identity cards. For example, use SSR and SNP molecular identity cards to compare known varieties to ensure that the molecular identity cards can accurately distinguish different germplasm resources of varieties.
[0060] (2) Optimization: According to the verification results, optimize the selection of molecular markers and amplification conditions. For example, for SSR markers, optimize the primer concentration and PCR reaction conditions to improve the specificity and stability of the amplification products; for SNP markers, optimize the chip hybridization and washing and staining conditions to improve the accuracy of genotyping.
[0061] 7. Classification management of germplasm resources: According to characteristics such as sample homozygosity and fingerprint specificity, the germplasm resources are divided into the following four categories: Core germplasm resources: Samples with relatively high homozygosity and SNP molecular identity card similarity < 95%, a total of 1,561 copies. Iso-related germplasm resources: Samples with highly similar molecular information (SNP molecular identity card similarity ≥ 95% between samples), a total of 705 copies. Heterogeneous germplasm resources: Samples with homozygosity below 90%, a total of 416 copies. Population germplasm resources: Genetic population germplasm resources for breeding or research needs, a total of 236 copies. Corresponding germplasm resource banks are established respectively to achieve classification management. For example, the core germplasm resource bank mainly stores pure-line germplasm resources with large genetic differences, and the iso-related germplasm resource bank mainly stores near-isogenic lines, similar varieties, and germplasm resources with homologous problems.
[0062] Example 2: Construction and application of molecular identity cards for osmanthus germplasm resource bank
[0063] 1. Collection of germplasm resource samples: Collect osmanthus samples of different varieties from the osmanthus germplasm resource bank, including tissues such as leaves and flowers. Each sample is numbered and recorded to ensure the traceability of the samples.
[0064] 2. DNA Extraction and Purification DNA was extracted from osmanthus samples using a plant DNA extraction kit. The concentration and purity of the DNA were detected by a NanoDrop spectrophotometer to ensure that the quality of the DNA met the requirements for subsequent analysis.
[0065] 3. Molecular Marker Selection and Amplification SSR marker primers suitable for osmanthus were selected, specific primers were designed, and PCR technology was used to amplify the target DNA fragments. The PCR reaction conditions, including annealing temperature, number of cycles, etc., were optimized to ensure the specificity and stability of the amplification products.
[0066] 4. Molecular Marker Detection and Analysis The PCR products were detected using a capillary electrophoresis instrument, and the sizes and numbers of alleles were analyzed. The detection results were organized into a data table, and the SSR marker information of each sample was recorded.
[0067] 5. Construction of Molecular Identity Cards Based on the combined characteristics of multiple SSR markers, a unique molecular identity card was constructed for each osmanthus variety. The molecular identity card can be in the form of digital coding, barcodes, or QR codes, etc., to integrate and represent the molecular marker information and associate it with the variety information to establish a database. Finally, verification was carried out.
[0068] The above is only the preferred embodiment of the present invention, and the protection scope of the present invention is not limited to the above embodiments. All technical solutions falling within the idea of the present invention belong to the protection scope of the present invention. It should be noted that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and refinements should also be regarded as within the protection scope of the present invention.
Claims
1. A method for constructing a molecular ID card of a germplasm resource bank and its application, characterized in that: The following steps are involved: Collect samples of target germplasm resources from the germplasm resource bank. Sample types include leaves, seeds and rhizomes of plants or tissues and blood of animals, and number and record the collected samples; Extract DNA from samples using a DNA extraction kit or a self-optimized extraction method, and test the quality and concentration of DNA by agarose gel electrophoresis and / or NanoDrop spectrophotometer to ensure the purity and integrity of the DNA; According to the type of target germplasm resources, select appropriate molecular marker types, including but not limited to single nucleotide polymorphisms (SNPs), simple sequence repeats (SSRs), and sequence feature amplification regions (AFLPs), design and select specific primers, use polymerase chain reaction (PCR) technology to amplify the target DNA fragments, and optimize PCR reaction conditions, including annealing temperature and cycle number, to ensure the specificity and stability of the amplified products; The amplified products were tested and the test results were analyzed by gel electrophoresis, capillary electrophoresis, and high-throughput sequencing to determine the molecular marker characteristics of each sample, including the type and number of alleles. The analysis results were organized into a data table to record the molecular marker information of each sample; Based on the combined characteristics of multiple molecular markers, a unique molecular ID card is constructed for each germplasm resource sample. The molecular ID card is in the form of digital code, barcode or QR code to integrate and represent the molecular marker information, and the molecular ID card is associated with other information of the germplasm resources to establish a database; The constructed molecular ID card is verified by comparing the molecular marker information of known samples to ensure its accuracy and uniqueness. Based on the verification results, the selection of molecular markers and amplification conditions are optimized to improve the reliability of the molecular ID card.
2. The method for constructing a molecular ID card of a germplasm resource bank and its application according to claim 1, characterized in that: During the sample collection step, for plant germplasm resources, select genetically stable and representative tissue sites for sampling, such as leaves, seeds, and rhizomes; for animal germplasm resources, select blood and tissue samples.
3. The method for constructing a molecular ID card of a germplasm resource bank and its application according to claim 1, characterized in that: In the DNA extraction step, corresponding extraction methods are used for different types of samples to improve the efficiency and quality of DNA extraction. For example, for plant leaf samples, a modified CTAB method is used to extract DNA; For animal blood samples, DNA was extracted using the phenol-chloroform method.
4. The method for constructing a molecular ID card of a germplasm resource bank and its application according to claim 1, characterized in that: In the molecular marker selection step, SSR markers are selected for plant germplasm resources; and SNP markers are selected for animal germplasm resources.
5. The method for constructing a molecular ID card of a germplasm resource bank and its application according to claim 1, characterized in that: In the molecular marker detection step, for SSR markers, capillary electrophoresis is used for detection; for SNP markers, high-throughput sequencing technology is used for detection.
6. The method for constructing a molecular ID card of a germplasm resource bank and its application according to claim 1, characterized in that: In the molecular ID card construction step, the molecular ID card is represented in the form of a two-dimensional code, which can efficiently integrate and store multiple molecular marker information and facilitate rapid reading and query through a scanning device.
7. The method for constructing a molecular ID card of a germplasm resource bank and its application according to claim 1, characterized in that: In the verification and optimization step, the molecular marker information of the known sample is compared to calculate the matching degree between the constructed molecular ID card and the known sample. When the matching degree is lower than the preset threshold, the selection and amplification conditions of the molecular marker are optimized to improve the accuracy and uniqueness of the molecular ID card.
8. Application of the method for constructing a molecular identity card of a germplasm resource bank according to any one of claims 1 to 7 in at least one of the following aspects: Germplasm resource identification and classification: Use molecular ID cards to quickly and accurately identify germplasm resources, distinguish different species, varieties or ecotypes, and provide a molecular basis for the classification and phylogenetic research of germplasm resources; Germplasm resource protection and management: Track the source and flow of germplasm resources through molecular ID cards to prevent misuse and loss of germplasm resources, provide a scientific basis for the management and maintenance of germplasm resource banks, and optimize the preservation conditions and strategies of germplasm resources; Utilization and development of germplasm resources: In the breeding process, use molecular IDs to screen germplasm resources with excellent traits, improve breeding efficiency, and provide technical support for the protection of new plant varieties and intellectual property protection; Biodiversity research: Provide molecular marker data for biodiversity research, analyze the genetic diversity and kinship of germplasm resources, and provide a scientific basis for the formulation of biodiversity conservation strategies.