Metabolic marker related to meat pigeon breast muscle rate and application thereof

Abstract

The application provides a kind of metabolic marker related to meat pigeon breast muscle rate and its application, belong to poultry genetics and molecular biology technical field.The metabolic marker is CDP-ethanolamine, and the relative content in the breast muscle tissue of high breast muscle rate individual is significantly higher than that of low breast muscle rate individual.The application screens the key difference metabolite by multi-omics joint analysis, and further carries out metabolite correlation whole genome analysis (mGWAS) by taking CDP-ethanolamine abundance as intermediate phenotype, and locates multiple genetic loci and candidate regulatory genes involved in phospholipid membrane synthesis pathway which are significantly related to it.The application not only provides precise biochemical target and reagent development basis for early evaluation and auxiliary breeding of high breast muscle rate individual of meat pigeon and other poultry, but also provides an analysis method for revealing the genetic basis of complex economic traits, has good cross-species stability and broad breeding application prospect.

Description

Technical Field

[0001] This invention belongs to the field of molecular metabolic regulation and bioinformatics analysis technology of pigeons, specifically relating to a metabolic biomarker related to the rate of pectoral muscle in pigeons and its application. Background Technology

[0002] Pigeons (Columba livia) are one of my country's important commercial meat-producing pigeons, with a long history of breeding. They are characterized by their small size, short breeding cycle, and strong adaptability. Pigeon meat is tender, with moderate fat content, rich in protein, and has high nutritional value. It is widely used in traditional dietary therapy and health care, and is gradually gaining popularity among consumers. With the expansion of the pigeon meat market and the improvement of consumption levels, the production performance of commercial meat pigeons, especially the yield and quality of breast muscle, has become an important criterion affecting economic benefits.

[0003] Currently, significant individual differences exist in pectoral muscle development among commercial pigeons of the same breed. These inter-individual differences directly affect meat yield and commercial value, but the molecular regulatory mechanisms are not yet fully understood. Although existing research focuses on the impact of breed differences or nutritional regulation on pigeon meat quality, systematic analytical methods for analyzing the pectoral muscle rate within the same breed are still lacking, especially studies combining metabolomics, genomics, and related pathway analyses, making it difficult to comprehensively reveal the key regulatory factors of pectoral muscle development.

[0004] Therefore, there is an urgent need for a method that can systematically analyze differences in pectoral muscle rate within the same breed to identify key metabolites and related genes closely related to pectoral muscle development, providing a scientific basis for breeding, meat quality optimization, and production performance improvement of commercial pigeons. Summary of the Invention

[0005] To address the issue that significant differences in breast muscle development among different individuals in pigeons directly affect meat yield and commercial value, but the molecular regulatory mechanism of breast muscle development in pigeons is not yet fully understood, this invention provides a metabolic biomarker related to breast muscle rate in pigeons and its application. This biomarker can distinguish individuals with high and low breast muscle rates within the same breed, revealing the molecular basis of breast muscle development in pigeons. It provides technical support for precision breeding and performance improvement of commercial pigeons, thus solving the aforementioned problems.

[0006] The technical solution of the present invention is as follows:

[0007] This invention provides a metabolic marker for evaluating or identifying the breast muscle rate trait of pigeons, wherein the metabolic marker is CDP-ethanolamine.

[0008] Preferably, the relative content of CDP-ethanolamine in the pectoral muscle tissue of individuals with high pectoral muscle percentage is significantly higher than that in individuals with low pectoral muscle percentage.

[0009] The present invention also provides the application of the above-mentioned metabolic marker CDP-ethanolamine in the preparation of a reagent for evaluating the development of breast muscles in pigeons or an auxiliary breeding tool.

[0010] Preferably, the auxiliary breeding tool is used to distinguish individuals with high and low pectoral muscle ratios within the same variety for molecular marker-assisted breeding.

[0011] The present invention also provides an auxiliary identification method for the pectoral muscle rate trait of pigeons, including detecting the content of CDP-ethanolamine in the isolated tissue sample of a candidate individual. If the content of CDP-ethanolamine in the isolated tissue sample of the candidate individual is higher than the reference value of individuals with low pectoral muscle rate, the candidate individual is determined to be an individual with high pectoral muscle rate dominance.

[0012] Preferably, the method further includes a joint evaluation by determining the content of other metabolites in the phospholipid membrane biosynthesis network, wherein the other metabolites include at least one of phosphatidylserine, phosphatidylethanolamine, and phosphatidylcholine.

[0013] This invention also provides an analytical method for the metabolic regulation mechanism related to the breast muscle rate of pigeons, comprising the following steps:

[0014] S1: Measure the individual pectoral muscle rate in a group of pigeons and divide them into a high pectoral muscle rate group and a low pectoral muscle rate group according to the pectoral muscle rate.

[0015] S2: Collect in vitro tissue samples from individuals and perform combined transcriptomic, proteomic, and metabolomic analyses to screen for common enrichment pathways across multiple omics between high and low pectoral muscle rate groups, and identify CDP-ethanolamine as the key metabolite corresponding to the common enrichment pathway.

[0016] S3: Using CDP-ethanolamine content as a phenotypic trait, conduct metabolite association genome-wide analysis to locate genomic loci and candidate regulatory genes significantly associated with CDP-ethanolamine.

[0017] Preferably, the metabolomics analysis in S2 is a non-target metabolomics analysis; the metabolite association genome-wide analysis in S3 uses a mixed linear model for association analysis.

[0018] Preferably, the candidate regulatory genes are mainly involved in the cell membrane phospholipid synthesis pathway.

[0019] Preferably, the candidate regulatory genes include at least one of GRIA1, MAPK12, EIF2AK3, WWP1, ACOX3, ABHD12B, ATP6V0D2, DOK7, SLC22A16, and PYGL; the expression levels of the candidate regulatory genes are significantly higher in individuals with high pectoral muscle mass than in individuals with low pectoral muscle mass.

[0020] The beneficial effects of this invention are as follows:

[0021] This invention uses the commercial white-feathered king pigeon as the research object, conducting research based on differences in pectoral muscle rate within the same breed. Through non-targeted metabolomics analysis combined with multi-omics co-enrichment of pathway information, the key metabolite CDP-ethanolamine closely related to pectoral muscle development was screened, and a metabolic network centered on phospholipid membrane synthesis metabolism was constructed. Further genome-wide association analysis of metabolite associations identified genetic loci and candidate genes significantly associated with CDP-ethanolamine content, with the located gene functions concentrated in phospholipid membrane synthesis and cellular structure regulation pathways. Subsequently, the metabolites related to the above-mentioned phospholipid membrane synthesis metabolism network were validated in Tarim pigeons, and the results showed that the metabolic network has consistency and stability across different pigeon breeds. This invention, through non-targeted metabolomics analysis, is the first to discover and validate CDP-ethanolamine as a metabolic marker for evaluating or identifying the pectoral muscle rate trait in pigeons. The relative content of this marker in the pectoral muscle tissue of individuals with high pectoral muscle rate is significantly higher than that of individuals with low pectoral muscle rate, effectively distinguishing between individuals with high and low pectoral muscle rates within the same breed, revealing the molecular basis of pectoral muscle development. This overcomes the technical bottleneck of difficulty in accurately quantifying phenotypic differences in the early stages of traditional breeding.

[0022] This invention provides a precise method for assisted breeding of pigeons based on CDP-ethanolamine abundance, using pectoral muscle percentage as an auxiliary factor. By further combining this method with the evaluation of other highly correlated metabolites in the phospholipid membrane biosynthesis network (such as phosphatidylserine, phosphatidylethanolamine, and phosphatidylcholine), the biosynthetic dependence is comprehensively reflected. This method improves the accuracy and reliability of validating superior individuals and selecting breeding stock, providing direct technical support for enhancing the production performance of commercial pigeons.

[0023] In traditional poultry breeding, yield phenotypes such as pectoral muscle percentage are regulated by multiple genes, making it difficult to directly identify key associated genes. This invention proposes a systematic method for metabolite association genome-wide analysis (mGWAS) that uses metabolite content as a phenotypic trait and combines it with genome-wide single nucleotide polymorphism (SNP) data. This method can not only accurately locate genetic loci significantly associated with CDP-ethanolamine phenotypic variations at the whole-genome level, but also further screen for candidate regulatory genes (such as GRIA1 and MAPK12) deeply involved in cell membrane phospholipid synthesis pathways. This approach extends traditional phenotypic observation to the metabolic and gene regulation levels, providing a practical and feasible means for early screening and breed optimization of muscle traits in pigeons. This method effectively simplifies the genetic analysis process of complex phenotypes and can accurately locate candidate genes and SNP loci involved in phospholipid membrane synthesis pathways and significantly affecting pectoral muscle development. The key gene and genetic locus information obtained through this analysis method provides clear molecular targets for the subsequent development of early gene screening technologies based on non-destructive samples such as blood and feathers. This not only overcomes the technical limitations of relying on slaughter to measure breast muscle or conduct quantitative sampling of metabolites in traditional breeding, but also provides a reliable underlying data foundation for molecular marker-assisted breeding (MAS) technology in poultry, which helps to shorten the breeding cycle of superior breeds and improve breeding efficiency.

[0024] The phospholipid membrane biosynthesis network and core biomarkers constructed using the method of this invention are also present in different pigeon breeds (Tarim pigeon population), and the related metabolites show strong correlation and synergistic effects. This verifies the high conservation, consistency, and stability of this metabolic pathway and related biomarkers in the regulation of avian pectoral muscle metabolism, ensuring the application potential and universality of the technical solution of this invention in the broad poultry breeding market. Attached Figure Description

[0025] Figure 1 : Pectoral muscle rate of high and low pectoral muscle rate groups of white-feathered king pigeons.

[0026] Figure 2 CDP-ethanolamine content and genome-wide association analysis. (A) Relative CDP-ethanolamine content in high and low pectoral muscle rate groups of white-feathered king pigeons; (B) QQ plot; (C) Manhattan plot.

[0027] Figure 3Candidate genes correspond to phospholipid membrane metabolic pathways. CDP: CDP-ethanolamine; PS: phosphatidylserine; PC: phosphatidylcholine; PE: phosphatidylethanolamine; DAG: diacylglycerol; SAH: S-adenosylhomocysteine; SAM: S-adenosylmethionine; EK: ethanolamine kinase; CK: choline kinase; ET: ethanolamine phosphorylated cytidine transferase; CT: choline phosphorylated cytidine transferase; EPT: ethanolamine phosphorylated phosphotransferase; CPT: choline phosphorylated phosphotransferase; PEMT: phosphatidylethanolamine N-methyltransferase; PTDSS: phosphatidylserine synthase; SER: serine.

[0028] Figure 4 CDP-ethanolamine content corresponding to genotype. (A) MAPK12; (B) ACOX3; (C) EIF2AK3; (D) FLVCR2; (E) PYGL; (F) ABHD12B; (G) ATP6V0D2-1; (H) ATP6V0D2-2; (I) WWP1; (J) DOK7; (K) SLC22A16; (L) PKD2.

[0029] Figure 5 Correlation of differentially metabolites in the phospholipid membrane metabolic network. CDP: CDP-ethanolamine; PS: phosphatidylserine; PC: phosphatidylcholine; PE: phosphatidylethanolamine; DAG: diacylglycerol; BMP: pectoral muscle rate.

[0030] Figure 6 Cross-species testing of the phospholipid membrane synergistic metabolic network. (A) Breast muscle rate of Tarim pigeons with high and low breast muscle rates; (B) Correlation of differentially metabolites in the phospholipid membrane metabolic network of Tarim pigeon breast muscle metabolites. PS: Phosphatidylserine; PC: Phosphatidylcholine; PE: Phosphatidylethanolamine; DAG: Diacylglycerol; BMP: Breast muscle rate. Detailed Implementation

[0031] The technical solution of the present invention will be described in detail below with reference to the embodiments and accompanying drawings. It should be understood that the following embodiments are for explanation and illustration only and are not intended to limit the scope of protection of the present invention.

[0032] In the following examples, reagents not specifically mentioned are all conventional reagents in the art, commercially available or prepared according to conventional methods in the art, and of laboratory purity grade. Vectors, cells, and experimental animals not specifically mentioned are all commercially available. Experimental methods and conditions not specifically mentioned are conventional experimental methods and conditions in the art, and can be found in relevant experimental manuals, publicly available literature, or manufacturer's instructions. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0033] The biological materials used in the following experimental examples:

[0034] The experimental animals used in the following examples were 28-day-old white-feathered king pigeons. Both the experimental animals and the experimental site were from Jiangsu Weitekai Pigeon Industry Co., Ltd. All experimental animals were raised from parent breeding pigeons to market weight. The parent breeding pigeons were fed a diet primarily composed of corn, soybeans, wheat, and sorghum to eliminate the influence of dietary factors on the breed's metabolites.

[0035] Unless otherwise specified, the reagents used in the following examples are all conventional reagents in the art, commercially available or prepared according to conventional methods in the art. Unless otherwise specified, the experimental methods and conditions used in the following examples are all conventional experimental methods and conditions in the art, and can be found in relevant experimental manuals, public literature, or manufacturer's instructions. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains.

[0036] Example 1. Grouping and Sample Acquisition Methods for Breast Muscle Ratio in Commercial Pigeons

[0037] The study used white-feathered king pigeons, commercial meat pigeons, as the research subjects, and raised them to slaughter age under unified feeding and management conditions. The body weight and breast muscle weight of each pigeon were measured, and the breast muscle percentage was calculated.

[0038] Based on the pectoral muscle percentage, individual white-feathered king pigeons were divided into a high pectoral muscle percentage group and a low pectoral muscle percentage group. The pectoral muscle percentages between the groups are as follows: Figure 1 As shown, there was a significant difference in pectoral muscle rate between the two groups (P = 9.36E-59), with 50 animals in each group.

[0039] Pectoral muscle tissue samples were collected rapidly after slaughter, flash-frozen in liquid nitrogen, and stored at −80°C for subsequent metabolite detection and molecular biological analysis.

[0040] Example 2. A method for screening key metabolites based on multi-omics pathway co-enrichment

[0041] Based on the pectoral muscle samples of the high and low pectoral muscle rate groups of white-feathered king pigeons described in Implementation Scheme 1, this implementation scheme is used to screen key metabolites that are significantly related to the difference in pectoral muscle rate. Specifically, it includes the following steps: (1) Multi-omics sample detection: Using the same batch of white-feathered king pigeon pectoral muscle samples as the research object, transcriptomics, proteomics and metabolomics detection are performed respectively to obtain the corresponding gene expression, protein abundance and metabolite quantitative information. (2) Differential feature screening: Genes, proteins and metabolites that show significant differences between the high and low pectoral muscle rate groups are screened at the transcriptomic, proteomic and metabolomic levels respectively. (3) Pathway-level integrated analysis: The differential genes, differential proteins and differential metabolites are annotated with functional pathways respectively, and the three types of differential features are integrated and analyzed at the pathway level to screen common biological pathways that show significant enrichment or regulatory trends at the transcriptional, protein and metabolic levels. (4) Identification of key metabolites: In the biological pathways that are commonly enriched, metabolites that show significant differences at the metabolome level were further screened, and CDP-ethanolamine was identified as a key metabolite closely related to the difference in breast muscle rate of white-feathered king pigeons (Table 1).

[0042] The test results show that ( Figure 2 The content of CDP-ethanolamine in the pectoral muscles of white-feathered king pigeons in the high pectoral muscle rate group was significantly higher than that in the low pectoral muscle rate group, and the corresponding phospholipid membrane synthesis-related pathways showed a consistent regulatory trend at the transcriptomic, proteomic, and metabolomic levels.

[0043] Table 1. Multi-omics co-enrichment pathways

[0044]

[0045] Example 3. Metabolite association whole-genome analysis method based on CDP-ethanolamine content

[0046] Based on the determination in Implementation Scheme 2 that CDP-ethanolamine is a key metabolite closely related to the difference in pectoral muscle rate in white-feathered king pigeons, this implementation scheme is used to analyze its genetic regulatory basis. Specifically, it includes the following steps: (1) Genomic data acquisition: Whole genome resequencing is performed on the white-feathered king pigeons used for metabolomics analysis in Implementation Scheme 1 to obtain high-quality single nucleotide polymorphism (SNP) data. (2) Phenotypic data construction: The CDP-ethanolamine content detected in the pectoral muscle samples of each individual is used as metabolic phenotypic data for subsequent association analysis. (3) Metabolite association genome-wide analysis (mGWAS): A mixed linear model is used to perform association analysis between CDP-ethanolamine content and whole genome SNP sites to control the influence of population structure and kinship on the analysis results, thereby obtaining genetic sites that are significantly associated with CDP-ethanolamine content. (4) Screening of significant association sites: According to the preset significance threshold, SNP sites that are significantly associated with CDP-ethanolamine content are screened to obtain multiple genome-wide significant association signals. (5) Candidate gene annotation: Functional annotation of the genomic regions where the significantly associated SNP sites are located or adjacent to them, and screening of candidate genes that may be involved in the regulation of phospholipid metabolism (Table 2).

[0047] The analysis results show that ( Figure 3 The candidate genes were mainly enriched in biological pathways related to phospholipid membrane synthesis, which is consistent with the analysis results of co-enrichment of multiple omics pathways in Implementation Scheme 2.

[0048] To elucidate the genetic basis of differences in CDP-ethanolamine content, this embodiment uses whole-genome resequencing data from a white-feathered king pigeon population and employs a mixed linear model to conduct a metabolite genome-wide association analysis (mGWAS), combined with QQ plots (…). Figure 2 The stability of the model was evaluated. The analysis results show that ( Figure 4 A total of 64 significant single nucleotide polymorphism (SNP) sites were identified at the whole-genome level (P < 8.94 × 10⁻⁶). -8 ) and 4,322 suggestive associated loci (P < 1.0 × 10⁻⁶). -6 Among them, the single site with the highest contribution explained 45.3% of the CDP-ethanolamine phenotypic variation, while the average explanation rates of significantly associated sites and suggestive sites were 29.6% and 23.9%, respectively.

[0049] After functional annotation of the aforementioned significantly associated loci, 44 potential candidate genes were identified. These genes mainly include EIF2AK3, PKD2, MAPK12, PYGL, ACOX3, FLVCR2, SLC22A16, ABHD12B, DOK7, WWP1, and ATP6V0D2. Further analysis of the genotype distribution of the associated loci revealed significant differences in the SNP genotype distribution of the aforementioned candidate genes between the high CDP-ethanolamine content group and the low CDP-ethanolamine content group (P < 0.05). Specifically, certain alleles located within the gene regions of PKD2, MAPK12, and ACOX3 showed an enrichment trend in individuals with high CDP-ethanolamine content, and this enrichment phenomenon was consistent with the distribution in individuals with high pectoral muscle mass.

[0050] Table 2 Summary of 44 candidate genes obtained by mGWAS

[0051]

[0052] Example 4. Methods for analyzing the phospholipid membrane biosynthetic network and its relationship with pectoral muscle rate.

[0053] Based on the non-target metabolomics data of white-feathered king pigeons obtained in Example 2, this example provides a method for constructing a phospholipid membrane biosynthesis network based on the same metabolomics data and analyzing its relationship with pectoral muscle rate. By performing pathway enrichment analysis on differential metabolites between the high and low pectoral muscle rate groups, phospholipid membrane biosynthesis-related metabolites were screened, with CDP-ethanolamine being the core differential metabolite. In the network analysis ( Figure 5CDP-ethanolamine was significantly positively correlated with phosphatidylserine (PS, r = 0.632, P = 0.0191), phosphatidylethanolamine (PE, r = 0.627, P = 0.0228), and phosphatidylcholine (PC, r = 0.612, P = 0.0267), indicating a synergistic effect in the membrane lipid biosynthesis network. However, it showed no correlation with diacylglycerol (DAG, r = 0.001, P = 0.995), suggesting its primary function is membrane lipid formation rather than energy storage. PS, PE, and PC were highly correlated (r > 0.98, P < 0.001), reflecting biosynthesis dependence. DAG was significantly correlated with pectoral muscle percentage (r = 0.337, P < 0.001), suggesting that energy lipids may also participate in muscle development. Further analysis showed that CDP-ethanolamine was moderately positively correlated with pectoral muscle percentage (r = 0.291, P = 0.003), and PS, PE, and PC were also significantly correlated with pectoral muscle percentage (PS: r = 0.215, P = 0.011; PE: r = 0.260, P = 0.009; PC: r = 0.204, P = 0.014), indicating that membrane structural lipids play a potential role in pectoral muscle development, while DAG independently participates in muscle growth. This method can be used to study key metabolic regulatory mechanisms related to pectoral muscle development in white-feathered king pigeons, providing a metabolic basis for optimizing pectoral muscle traits.

[0054] Example 5. Detection of gene expression related to phospholipid membrane biosynthesis network

[0055] Based on the phospholipid membrane anabolic metabolic network screened in Example 4, this example provides a method for detecting gene expression in the phospholipid membrane anabolic metabolic network. According to the differences in CDP-ethanolamine expression between high and low pectoral muscle rate groups in Example 4, total RNA was extracted from pectoral muscle tissue samples, and the expression levels of candidate genes related to CDP-ethanolamine metabolism were detected using real-time quantitative PCR (qPCR). Primer sequences are shown in Table 3. The detection results (Table 4) show that the relative expression levels of candidate genes (GRIA1, MAPK12, EIF2AK3, WWP1, ACOX3, ABHD12B, ATP6V0D2, DOK7, SLC22A16, PYGL) were significantly higher in the high pectoral muscle rate group than in the low pectoral muscle rate group, verifying the potential regulatory role of the phospholipid membrane anabolic metabolic network in pectoral muscle development. This method can be used to further elucidate the mechanism of action of the phospholipid membrane anabolic metabolic network in pectoral muscle formation and growth, providing a molecular basis for improving the pectoral muscle traits of white-feathered king pigeons.

[0056] Table 3 Primer sequences for candidate genes

[0057]

[0058] Table 4. Gene expression related to the phospholipid membrane biosynthesis network in the pectoral muscle of White King Pigeon

[0059]

[0060] Note: Different letters a and b indicate significant differences between groups (P < 0.05).

[0061] Example 6. Validation of metabolites related to the phospholipid membrane biosynthesis network in Tarim pigeons

[0062] To verify the applicability of the phospholipid membrane biosynthesis network-related metabolites screened in Example 4 to another pigeon species, this example selected pectoral muscle tissue from 50 Tarim pigeons for testing. The contents of metabolites such as CDP-ethanolamine, PS, PC, PE, and DAG were determined using non-target metabolomics methods, and the correlation of these metabolites in the pectoral muscle tissue was analyzed. The results showed ( Figure 6 Strong correlations were also observed among PS, PC, PE, and DAG (PS vs. PC: r = 0.10; PS vs. PE: r = 0.44; PC vs. PE: r = 0.23; DAG vs. PE: r = 0.20; BMP vs. DAG: r = 0.49), reflecting their synergistic effects in the phospholipid membrane biosynthesis network. These results indicate that the phospholipid membrane biosynthesis network established in Example 4 also exists in Tarim pigeons, verifying the conservation of metabolites related to this pathway and their role in regulating pectoral muscle development, providing experimental evidence for further research.

Claims

1. A metabolic marker for evaluating or identifying the breast muscle rate trait in pigeons, characterized in that, The metabolic marker used to evaluate or identify the breast muscle rate trait of pigeons is CDP-ethanolamine.

2. The metabolic marker for evaluating or identifying the breast muscle rate trait of pigeons according to claim 1, characterized in that, The relative content of CDP-ethanolamine in the pectoral muscle tissue of individuals with high pectoral muscle percentage was significantly higher than that in individuals with low pectoral muscle percentage.

3. The application of the metabolic markers for evaluating or identifying the breast muscle rate trait of pigeons as described in claim 1 or 2 in the preparation of evaluation reagents for breast muscle development in pigeons or auxiliary breeding tools.

4. The application according to claim 3, characterized in that, The auxiliary breeding tool is used to distinguish individuals with high and low pectoral muscle ratios within the same variety for molecular marker-assisted breeding.

5. An auxiliary identification method for the pectoral muscle percentage trait of pigeons, characterized in that, This includes detecting the CDP-ethanolamine content in ex vivo tissue samples of candidate individuals. If the CDP-ethanolamine content in the ex vivo tissue samples of a candidate individual is higher than the reference value for individuals with low pectoral muscle percentage, then the candidate individual is determined to be an individual with high pectoral muscle percentage dominance.

6. The auxiliary identification method according to claim 5, characterized in that, The auxiliary identification method includes a joint evaluation by measuring the content of other metabolites in the phospholipid membrane biosynthesis network, wherein the other metabolites include at least one of phosphatidylserine, phosphatidylethanolamine and phosphatidylcholine.

7. An analytical method for the metabolic regulatory mechanism related to the breast muscle rate of pigeons, characterized in that, Includes the following steps: S1: Measure the individual pectoral muscle rate in a group of pigeons and divide them into a high pectoral muscle rate group and a low pectoral muscle rate group according to the pectoral muscle rate. S2: Collect in vitro tissue samples from individuals and perform combined transcriptomic, proteomic, and metabolomic analyses to screen for common enrichment pathways across multiple omics between high and low pectoral muscle rate groups, and identify CDP-ethanolamine as the key metabolite corresponding to the common enrichment pathway. S3: Using CDP-ethanolamine content as a phenotypic trait, conduct metabolite association genome-wide analysis to locate genomic loci and candidate regulatory genes significantly associated with CDP-ethanolamine.

8. The analytical method according to claim 7, characterized in that, The metabolomics analysis in S2 is a non-target metabolomics analysis; the metabolite association genome-wide analysis in S3 uses a mixed linear model for association analysis.

9. The analytical method according to claim 8, characterized in that, The candidate regulatory genes in S3 are mainly involved in the cell membrane phospholipid synthesis pathway.

10. The analytical method according to claim 9, characterized in that, The candidate regulatory genes include at least one of GRIA1, MAPK12, EIF2AK3, WWP1, ACOX3, ABHD12B, ATP6V0D2, DOK7, SLC22A16, and PYGL; the expression levels of the candidate regulatory genes are significantly higher in individuals with high pectoral muscle mass than in individuals with low pectoral muscle mass.

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