IncRNA (long non-coding ribonucleic acid) related to chicken breast muscle development character and application thereof

Abstract

The invention discloses lncRNA (long non-coding ribonucleic acid) related to chicken breast muscle development traits and application of the lncRNA. The expression levels of lncRNA and mRNA are detected through a next-generation sequencing technology, and the lncRNA which affects the expression of a key factor MYOD1 gene for chicken development is found on the basis of lncRNA-mRNA association analysis, and the lncRNA is lncRNA MSTRG.14589.1. Further, molecular biology test verification is carried out in expanded groups and different varieties, and the result shows that the lncRNA MSTRG.14589.1 is obviously differentially expressed in individuals with different pectoral muscle weights; the lncRNA MSTRG.14589.1 is in remarkable positive correlation with MYOD1, so that the muscle development of the chicken is influenced. The lncRNA provided by the invention can be used as a molecular marker related to evaluation of chicken muscle development potential, and has important practical application value in molecular breeding or muscle development regulation.

Description

Technical Field

[0001] The present invention belongs to the technical field of chicken molecular breeding, and in particular, relates to a lncRNA related to chicken breast muscle development traits and an application thereof. Background Art

[0002] Chicken is the world's most consumed meat, and chicken production accounts for more than 1 / 3 of the total meat production. In my country, chicken is the second largest meat resource, and its consumption proportion is increasing year by year. Compared with large animals such as pigs and cattle, white-feathered broilers have the advantages of high feed conversion efficiency, low unit protein price, and low carbon emissions. They are typical environmentally friendly agricultural animals. In addition, white-feathered broilers have a shorter hatching and breeding cycle, more flexible production adjustment, and a more complete upstream and downstream industrial chain, which can quickly adapt to changing market demands. Breast muscle traits are an important indicator for measuring the quality of broiler meat production performance. Affected by factors such as avian influenza, live poultry trading is strictly regulated. In a market environment dominated by chilled fresh chickens and cut chickens, developed breast muscles can shape broilers into a fuller shape, which is conducive to increasing consumer interest and willingness to buy. Therefore, breast muscle traits have always been the most concerned phenotype in broiler breeding.

[0003] Skeletal muscle is the most important part of the poultry body. Its basic structure is mainly muscle fiber. Therefore, the development of muscle fiber is closely related to the production efficiency and meat quality of poultry. Generally speaking, the development of muscle fiber can be divided into two stages: the first stage is the formation of primary muscle fiber, which is formed by the directed development of mesenchymal stem cells in the myogenic node into myoblasts, which is usually completed from the 5th to the 7th day of the embryonic period; the second stage is the formation of secondary muscle fiber, in which myotubes are formed and differentiated into mature muscle fibers, which is usually completed from the 16th day of the embryonic period to hatching. During the formation and maturation of myotubes, unfused single myoblasts are isolated between the endomysium to form muscle satellite cells, which play an important role in the repair process of animal muscle damage. In the embryonic period, the number of poultry muscle fibers is basically fixed. The development of muscle fibers at this stage determines the potential for muscle development after the animal is born.

[0004] With the development of sequencing technology and molecular biology, people's understanding of the regulatory mechanism of muscle development is no longer limited to coding genes. The understanding of the regulatory mechanism of lncRNA related to pectoral muscle development has gradually deepened, and more and more lncRNAs have been found to play a role in the growth and development of chicken pectoral muscle. The chicken muscle-specific expression of lncRNA MYH1G-AS is synergistically regulated by SP2 and SMAD3, which can promote myoblast proliferation, inhibit myoblast differentiation, and promote the conversion of slow muscle fibers to fast muscle fibers, thereby causing muscle atrophy. lncRNA plays a wide range of regulatory roles in muscle development, but at present, only a small number of lncRNA mechanisms have been verified, and there is still a large research area to be explored.

[0005] In the current environment, chicken breast muscle traits have received widespread attention in scientific research and consumer markets. It is necessary to explore the epigenetic molecular markers and target genes that affect breast muscle development, determine the development potential of chicken breast muscle, and then assist in broiler molecular breeding, ultimately improving production efficiency. Summary of the invention

[0006] The purpose of the present invention is to provide a lncRNA related to chicken breast muscle development traits and an application thereof.

[0007] In order to achieve the purpose of the present invention, in a first aspect, the present invention provides a lncRNA (MSTRG.14589.1) related to chicken breast muscle development traits, and its corresponding nucleotide sequence is shown in SEQ ID NO:1.

[0008] In a second aspect, the present invention provides specific primers for detecting the lncRNA, including an upstream primer as shown in SEQ ID NO: 2 and a downstream primer as shown in SEQ ID NO: 3.

[0009] In a third aspect, the present invention provides a detection reagent or a kit containing the primers.

[0010] The present invention also provides a fluorescent quantitative PCR kit for detecting the expression level of lncRNA MSTRG.14589.1, which contains all reagents required for the whole set of total RNA extraction, reverse transcription, and qPCR test processes, such as reverse transcription reagents, reverse transcription primers, specific upstream primers, specific downstream primers, internal reference primers (for example, RPL32 and HSPA2 are used as internal reference genes, and the NCBI numbers are 416122 and 423504, respectively), and fluorescent quantitative PCR reaction solution. The specific primers of the two internal reference genes are: RPL32 upstream primer: 5'-AGTTCATCCGCCACCAGTCTGAT-3'; downstream primer: 5'-GCTTCGTCTTCTTGTTGCTCCCATA-3'; HSPA2 upstream primer: 5'-TCTGCTCCTGTTGGATGTC-3'; downstream primer: 5'-TGGGAATGGTGGTGTTACG-3'.

[0011] In a fourth aspect, the present invention provides any of the following applications of the lncRNA, the primer, or the detection reagent or kit: (1) Used for the breeding of chicken breast muscle development traits; (2) Used for early prediction of high-yield broiler performance chicken individuals; (3) Used in chicken molecular marker-assisted breeding; (4) Used for genetic improvement of chicken breeds; (5) Used to predict the expression level of chicken MYOD1 gene; the reference sequence number of chicken MYOD1 gene in NCBI is 374048.

[0012] lncRNA MSTRG.14589.1 was significantly positively correlated with MYOD1 and affected chicken muscle development.

[0013] In a fifth aspect, the present invention provides a method for breeding chicken breast muscle development traits, the method comprising: detecting the transcription abundance of the lncRNA as shown in SEQ ID NO: 1 in the muscle tissue of the chicken to be tested, the chickens with low transcription abundance are high breast muscle individuals, and conversely, the chickens with high transcription abundance are low breast muscle individuals.

[0014] The lncRNA of the present invention is negatively correlated with muscle development. The lncRNA transcription abundance is detected by qPCR technology, and selection is performed based on the relative expression level. When the expression level difference reaches more than 20%, low-breast muscle chicken individuals (lncRNA high-expression individuals) and high-breast muscle chicken individuals (lncRNA low-expression individuals) can be distinguished, and only low-expression individuals need to be selected.

[0015] Furthermore, the method comprises the following steps: 1) Extract total RNA from chicken breast muscle tissue to be tested, and reverse transcribe to synthesize cDNA; 2) Using cDNA as a template, PCR amplification was performed using primers shown in SEQ ID NO: 2-3; 3) Analyze the PCR amplification products.

[0016] Preferably, the PCR reaction conditions used are: 95°C for 30s; 95°C for 3s, 60°C for 34s, for 40 cycles.

[0017] By means of the above technical solution, the present invention has at least the following advantages and beneficial effects: The lncRNA MSTRG.14589.1 provided by the present invention is highly correlated with chicken breast muscle development traits and can be used as a molecular marker in the genetic breeding of chicken meat production traits.

[0018] The fluorescent quantitative PCR kit for detecting the expression level of lncRNA MSTRG.14589.1 provided by the present invention is easy to use, simple to operate, and has high consistency in results. BRIEF DESCRIPTION OF THE DRAWINGS

[0019] Figure 1 This is to expand the phenotypic differences of individual groups of high and low pectoral muscles in a population in a preferred embodiment of the present invention.

[0020] Figure 2In a preferred embodiment of the present invention, the expression differences of MSTRG.14589.1 and MYOD1 in high and low pectoral muscle weight groups are expanded.

[0021] Figure 3 In a preferred embodiment of the present invention, the correlation between the pectoral muscle lncRNA MSTRG.14589.1 and MYOD1 is expanded.

[0022] Figure 4 This is a preferred embodiment of the present invention to expand the correlation between the pectoral muscle lncRNA MSTRG.14589.1 and the pectoral muscle weight of the population.

[0023] Figure 5 This is a preferred embodiment of the present invention to expand the correlation between the pectoral muscle lncRNA MSTRG.14589.1 and the pectoral muscle rate in the population.

[0024] Figure 6 This is the phenotypic difference of the high and low breast muscles of the three yellow chickens in the preferred embodiment of the present invention.

[0025] Figure 7 This is the difference in expression of MSTRG.14589.1 and MYOD1 between the high and low breast muscle weight groups of three yellow chickens in a preferred embodiment of the present invention.

[0026] Figure 8 This is the correlation between lncRNA MSTRG.14589.1 and MYOD1 in the pectoral muscle of three-yellow chicken in a preferred embodiment of the present invention.

[0027] Fig. 9 This is the correlation between the pectoral muscle lncRNA MSTRG.14589.1 and the pectoral muscle weight of the three-yellow chicken in a preferred embodiment of the present invention.

[0028] Fig.10 This is the correlation between the pectoral muscle lncRNA MSTRG.14589.1 and the pectoral muscle rate of the three-yellow chicken in a preferred embodiment of the present invention.

[0029] Fig.11 This is the comparison result of breast muscle weight and breast muscle rate between AA chicken and Xiaoshan chicken in the preferred embodiment of the present invention.

[0030] Fig.12 This is the comparison result of lncRNA MSTRG.14589.1 and MYOD1 expression levels between AA chicken and Xiaoshan chicken in the preferred embodiment of the present invention.

[0031] DETAILED DESCRIPTION

[0032] The present invention detects the expression levels of lncRNA and mRNA through the second generation sequencing technology, and finds a lncRNA that affects the expression of the MYOD1 gene, a key factor in chicken meat development, based on the lncRNA-mRNA association analysis, which is lncRNAMSTRG.14589.1. Further molecular biological experiments were conducted in the expanded population and different breeds to verify that lncRNA MSTRG.14589.1 was significantly differentially expressed in individuals with different breast muscle weights; lncRNA MSTRG.14589.1 was significantly positively correlated with MYOD1, affecting chicken muscle development.

[0033] The LncRNA MSTRG.14589.1 marker determination method is as follows: 1) Total RNA from chicken breast muscle tissue was extracted using TRIzol reagent; 2) Reverse transcription using FastKing cDNA First Strand Synthesis Kit; 3) qPCR experiments were performed using lncRNA MSTRG.14589.1-specific upstream and downstream primers and the KAPA SYBR FASTUniversal qPCR Kit.

[0034] The present invention also provides a detection kit for detecting the above-mentioned lncRNA markers and / or detecting the target gene MYOD1. The PCR kit is suitable for all types of fluorescent quantitative gene amplification instruments currently on the market, with high sensitivity, rapid and accurate quantification, good stability, and good application prospects. A fluorescent quantitative PCR kit for detecting the expression level of lncRNA MSTRG.14589.1 provided by the present invention includes: a reverse transcription reagent, a reverse transcription primer, a specific upstream primer, a specific downstream primer, an internal reference primer, and a fluorescent quantitative PCR reaction solution. The specific upstream and downstream primer sequences are shown in SEQ ID NOs: 2 and 3, respectively.

[0035] The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention. Unless otherwise specified, the technical means used in the examples are conventional means well known to those skilled in the art, and the raw materials used are all commercially available products.

[0036] Example 1 Obtaining lncRNA and key genes related to chicken breast muscle weight and breast muscle rate using high-throughput sequencing technology

[0037] 1) Experimental animals and phenotypic determination

[0038] Xianju chicken (140 days old) was used as the experimental material. 30 hens were randomly selected, weighed and slaughtered after fasting for 12 hours. The right pectoral muscle tissue was collected and quick-frozen in liquid nitrogen. The right pectoral muscle was then peeled off and weighed, and the pectoral muscle rate was calculated.

[0039] 2) Identification of differentially expressed lncRNAs and genes based on high-throughput sequencing results

[0040] Based on the phenotype of chest muscle weight and chest muscle rate, high and low chest muscle phenotypes were grouped (n=6), and lncRNA and transcriptome sequencing was performed on the chest muscle tissues of individuals in the high and low chest muscle groups. The original data after quality control were subjected to genome alignment and transcript analysis. On the one hand, the transcription abundance of known lncRNAs was annotated, and on the other hand, new lncRNAs were annotated using CPC2, CNCI, and Freelnc software. The transcript length>200 bp, exon data≥2, and the results identified as lncRNA by the three softwares were retained. The expression of lncRNA and genes was corrected using DESeq2 software, and the differential expression fold>1.5 and p value<0.05 were defined as the significant differential expression standards to identify differentially expressed lncRNAs and differentially expressed genes.

[0041] 3) Joint analysis of differentially expressed lncRNAs and genes for key lncRNA screening

[0042] The cis target gene analysis of differentially expressed lncRNA was performed. The location information of differentially expressed lncRNA was extracted, and genes within 100 kb upstream and downstream were mined and defined as cis target genes. The target genes were annotated and enriched, and candidate genes related to muscle development were screened. Genes such as MYOD1, SMAD1, and TPM1 were identified. The correlation analysis between cis target genes and lncRNA was performed, and R 2 >0.6 was used as the standard to filter candidate target genes and lncRNAs, and the correlation analysis between lncRNA and phenotype was performed. Finally, it was found that lncRNA MSTRG.14589.1 potentially regulates MYOD1 and affects muscle development (Table 1).

[0043] The above results show that lncRNA MSTRG.14589.1 is differentially expressed in individuals with high and low pectoral muscle groups and is significantly correlated with the pectoral muscle phenotype. MYOD1 is an important functional gene that affects muscle development and is regulated by lncRNA MSTRG.14589.1, indicating that lncRNA MSTRG.14589.1 can participate in the regulation of muscle development by affecting MYOD1 expression.

[0044] Table 1 Candidate lncRNAs and target genes

[0045] Example 2 Verification of the correlation between lncRNA MSTRG.14589.1 and MYOD1 and breast muscle phenotype using the Xianju chicken population

[0046] 1) Test sample preparation

[0047] A group of 140-day-old Xianju chickens (n=60) were used as the experimental material, and the sampling process was the same as in Example 1. The phenotypes such as body weight and breast muscle weight were recorded, and the breast muscle rate was calculated. The breast muscle tissue samples were collected, quickly frozen in liquid nitrogen, and stored.

[0048] 2) Verification of lncRNA MSTRG.14589.1 and MYOD1 expression

[0049] Total RNA from pectoral muscle tissue was extracted using TRIzol and reverse transcribed using FastKing cDNA First-Strand Synthesis Kit (TIANGEN, China), and the cDNA concentration was uniformly adjusted. qPCR experiments were performed under the following conditions: lncRNA MSTRG.14589.1 qPCR reaction conditions: upstream and downstream primers F: 5'-CACAAGAGGATGGAAGCCAGA-3', R: 5'-TGACTTGGGTTTGTGGTCCT-3'. KAPA SYBR FASTUniversal qPCR Kit (KAPA, USA) was used to configure a 10 ul qPCR reaction system, with 3 technical replicates per sample, and RPL32 and HSPA2 were used as internal reference genes. qPCR working procedure: 95℃ 30s; 95℃ 3s, 60℃ 34s (40 replicates).

[0050] MYOD1 qPCR reaction conditions: upstream and downstream primers F: 5'-GCTACTACACGGAATCACCAAAT-3', R: 5'-CTGGCTCCACTGTCACTCA-3'. KAPA SYBR FAST Universal qPCR Kit (KAPA, USA) was used to configure a 10 ul qPCR reaction system, with 3 technical replicates per sample, and RPL32 and HSPA2 were used as internal reference genes. qPCR working procedure: 95℃ 30s; 95℃ 3s, 60℃ 34s (40 replicates).

[0051] Calculation of chest muscle rate: Calculate based on different individual body weight and chest muscle weight. The calculation formula is:

[0052] Association analysis between lncRNA MSTRG.14589.1 and MYOD1 and phenotypic indicators: Pearson correlation analysis was performed on the expression level of muscle lncRNA MSTRG.14589.1 and MYOD1, as well as pectoral muscle weight and pectoral muscle rate, based on the linear regression calculation method. p<0.05 was used as the significance threshold. The analysis was performed using SPSS 25.0 software (SPSS, Chicago, IL, USA).

[0053] In the expanded group (n=60), high and low pectoral muscle weight groups (n=15) ( Figure 1 ), firstly, the expression differences of lncRNA MSTRG.14589.1 and MYOD1 in high and low pectoral muscle reconstructed individuals were compared. qPCR results showed that the expression levels of lncRNA MSTRG.14589.1 and MYOD1 in the muscles of high pectoral muscle reconstructed individuals were significantly reduced ( Figure 2 ); Based on the expression analysis of all individuals (n=60), it was found that lncRNA MSTRG.14589.1 was significantly positively correlated with MYOD1 (R 2 =0.454, p<1.68E-08)( Figure 3 and lncRNA MSTRG.14589.1 was associated with pectoral muscle weight (R 2 =0.147, p<0.01), chest muscle rate (R 2 =0.181, p<0.01) was significantly negatively correlated ( Figure 4 and Figure 5 ).

[0054] Example 3 Verification of the correlation between lncRNA MSTRG.14589.1 and MYOD1 and phenotype in breast muscle of three-yellow chicken population

[0055] 1) Test sample preparation

[0056] 98-day-old three-yellow chickens (n=40) were used as test materials. The sampling process was the same as in Example 1. 40 hens were randomly selected, weighed and slaughtered after fasting for 12 hours. The right breast muscle tissue was collected and quick-frozen in liquid nitrogen. The right breast muscle was peeled off and weighed to calculate the breast muscle rate.

[0057] 2) Verification of lncRNA MSTRG.14589.1 and MYOD1 expression

[0058] The method is the same as in Example 2, specifically as follows

[0059] Total RNA from pectoral muscle tissue was extracted using TRIzol and reverse transcribed using FastKing cDNA First-Strand Synthesis Kit (TIANGEN, China), and the cDNA concentration was uniformly adjusted. qPCR experiments were performed under the following conditions: lncRNA MSTRG.14589.1 qPCR reaction conditions: upstream and downstream primers F: 5'-CACAAGAGGATGGAAGCCAGA-3', R: 5'-TGACTTGGGTTTGTGGTCCT-3'. KAPA SYBR FASTUniversal qPCR Kit (KAPA, USA) was used to configure a 10 ul qPCR reaction system, with 3 technical replicates per sample, and RPL32 and HSPA2 were used as internal reference genes. qPCR working procedure: 95℃ 30s; 95℃ 3s, 60℃ 34s (40 replicates).

[0060] MYOD1 qPCR reaction conditions: upstream and downstream primers F: 5'-GCTACTACACGGAATCACCAAAT-3', R: 5'-CTGGCTCCACTGTCACTCA-3'. KAPA SYBR FAST Universal qPCR Kit (KAPA, USA) was used to configure a 10 ul qPCR reaction system, with 3 technical replicates per sample, and RPL32 and HSPA2 were used as internal reference genes. qPCR working procedure: 95℃ 30s; 95℃ 3s, 60℃ 34s (40 replicates).

[0061] Calculation of chest muscle rate: Calculate based on different individual body weight and chest muscle weight. The calculation formula is:

[0062] Association analysis between lncRNA MSTRG.14589.1 and MYOD1 and phenotypic indicators: Pearson correlation analysis was performed on the expression level of muscle lncRNA MSTRG.14589.1 and MYOD1, as well as pectoral muscle weight and pectoral muscle rate, based on the linear regression calculation method. p<0.05 was used as the significance threshold. The analysis was performed using SPSS 25.0 software (SPSS, Chicago, IL, USA).

[0063] In the three-yellow chicken group (n=40), high and low breast muscle weights were grouped (n=15). First, the expression differences of lncRNA MSTRG.14589.1 and MYOD1 in high and low breast muscle recombinant individuals were compared ( Figure 6). qPCR results showed that the expression levels of lncRNA MSTRG.14589.1 and MYOD1 in the reconstructed high pectoral muscles were significantly reduced ( Figure 7 ); Based on the expression analysis of all individuals (n=40), it was found that lncRNA MSTRG.14589.1 was significantly positively correlated with MYOD1 (R 2 =0.536, p<1.39E-05)( Figure 8 and lncRNA MSTRG.14589.1 was associated with pectoral muscle weight (R 2 =0.149, p<0.05), chest muscle rate (R 2 =0.273, p<0.001) was significantly negatively correlated ( Fig. 9 and Fig.10 ).

[0064] Example 4 Verification of the correlation between lncRNA MSTRG.14591.1 and MYOD1 and phenotype in breast muscle of fast-growing white-feathered broilers and Xiaoshan chickens

[0065] 1) Test sample preparation

[0066] One-day-old AA broiler chickens and Xiaoshan chickens (n ​​= 30) were purchased and raised and fed with the same diet in the same environment. The sampling process was the same as in Example 1. At 42 days of age, 15 chickens were randomly selected and weighed after fasting for 12 hours and slaughtered. The right breast muscle tissue was collected and quick-frozen in liquid nitrogen. The right breast muscle was peeled off and weighed to calculate the breast muscle rate.

[0067] 2) Verification of lncRNA MSTRG.14589.1 and MYOD1 expression

[0068] The method is the same as in Example 2, specifically as follows: Total RNA from pectoral muscle tissue was extracted using TRIzol and reverse transcribed using FastKing cDNA First-Strand Synthesis Kit (TIANGEN, China), and the cDNA concentration was uniformly adjusted. qPCR experiments were performed under the following conditions: lncRNA MSTRG.14589.1 qPCR reaction conditions: upstream and downstream primers F: 5'-CACAAGAGGATGGAAGCCAGA-3', R: 5'-TGACTTGGGTTTGTGGTCCT-3'. KAPA SYBR FASTUniversal qPCR Kit (KAPA, USA) was used to configure a 10 ul qPCR reaction system, with 3 technical replicates per sample, and RPL32 and HSPA2 were used as internal reference genes. qPCR working procedure: 95℃ 30s; 95℃ 3s, 60℃ 34s (40 replicates).

[0069] MYOD1 qPCR reaction conditions: upstream and downstream primers F: 5'-GCTACTACACGGAATCACCAAAT-3', R: 5'-CTGGCTCCACTGTCACTCA-3'. KAPA SYBR FAST Universal qPCR Kit (KAPA, USA) was used to configure a 10 ul qPCR reaction system, with 3 technical replicates per sample, and RPL32 and HSPA2 were used as internal reference genes. qPCR working procedure: 95℃ 30s; 95℃ 3s, 60℃ 34s (40 replicates).

[0070] Calculation of chest muscle rate: Calculate based on different individual body weight and chest muscle weight. The calculation formula is:

[0071] Association analysis between lncRNA MSTRG.14589.1 and MYOD1 and phenotypic indicators: The t-test was used to compare the differences in breast muscle phenotypes between AA chickens and Xiaoshan chickens, and the differences in the expression levels of lncRNA MSTRG.14591.1 and MYOD1, with p < 0.05 as the significance threshold. The analysis was performed using SPSS 25.0 software (SPSS, Chicago, IL, USA).

[0072] The breast muscle weight and breast muscle rate of AA broiler chickens were significantly higher than those of Xiaoshan chickens ( Fig.11 ), while the expression levels of lncRNAMSTRG.14591.1 and MYOD1 in breast muscle of AA chicken were significantly lower than those in Xiaoshan chicken ( Fig.12 ). This indicates that the regulation of lncRNAMSTRG.14591.1 and MYOD1 on breast muscle traits still holds true among different breeds.

[0073] Example 5 A detection kit for lncRNA MSTRG.14589.1 related to chicken breast muscle development potential

[0074] Extraction of muscle lncRNA: Total RNA from muscle tissue was extracted using TRIzol reagent.

[0075] The detection kit prepared in this example includes: a reverse transcription reagent, a reverse transcription primer, a specific upstream primer, a specific downstream primer, an internal reference primer (with RPL32 and HSPA2 as internal reference genes), and a fluorescent quantitative PCR reaction solution.

[0076] The reagents and primers used for reverse transcription and qPCR reactions are as follows:

[0077] Although the present invention has been described in detail above with general descriptions and specific embodiments, it is obvious to those skilled in the art that some modifications or improvements can be made based on the present invention. Therefore, these modifications or improvements made without departing from the spirit of the present invention all fall within the scope of protection claimed by the present invention.

Claims

1. A lncRNA associated with chicken breast muscle development traits, characterized in that: The corresponding nucleotide sequence is shown in SEQ ID NO:

1.

2. A specific primer for detecting the lncRNA according to claim 1, characterized in that: It includes an upstream primer as shown in SEQ ID NO:2 and a downstream primer as shown in SEQ ID NO:

3.

3. A detection reagent or kit containing the primers according to claim 2.

4. Any of the following uses of the lncRNA according to claim 1, the primer according to claim 2, or the detection reagent or kit according to claim 3: (1) Used for the breeding of chicken breast muscle development traits; (2) Used for early prediction of high-yield broiler performance chicken individuals; (3) Used in chicken molecular marker-assisted breeding; (4) Used for genetic improvement of chicken breeds; (5) Used to predict the expression level of chicken MYOD1 gene; The reference sequence number of the chicken MYOD1 gene at NCBI is 374048.

5. A method for breeding chicken breast muscle development traits, characterized in that: The method comprises: detecting the transcription abundance of the lncRNA shown in SEQ ID NO: 1 in the muscle tissue of the tested chicken, wherein the chickens with low transcription abundance are high-breast muscle chickens, and conversely, the chickens with high transcription abundance are low-breast muscle chickens.

6. The method according to claim 5, characterized in that The following steps are involved: 1) Extract total RNA from chicken breast muscle tissue to be tested, and reverse transcribe to synthesize cDNA; 2) Using cDNA as a template, PCR amplification was performed using primers shown in SEQ ID NO: 2-3; 3) Analyze the PCR amplification products.

7. The method according to claim 6, characterized in that The PCR reaction conditions used were: 95°C for 30 s; 95°C for 3 s, 60°C for 34 s, for 40 cycles.

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