A gene combination related to body shape of large yellow croaker and application thereof in breeding

Abstract

This invention discloses a gene combination related to the body shape of large yellow croaker and its application in breeding, belonging to the field of fish fry and fingerling reproduction technology. The gene combination related to the body shape of large yellow croaker includes the INHBB gene, Nodal gene, GDF3 gene, ACVR2A gene, ALK4 gene, and Smad2 gene. This invention is applied to the screening of large yellow croaker body shape, providing molecular markers for large yellow croaker breeding screening.

Description

Technical Field

[0001] This invention belongs to the field of fish fry and fingerling breeding technology, and in particular relates to a gene combination related to the body shape of large yellow croaker and its application in breeding. Background Technology

[0002] The depletion of global fishery resources has prompted a shift from traditional fishing to aquaculture. Marine aquaculture has become a key method for sustainable seafood production. China, as the world's largest aquaculture producer, has taken the lead in this transformation, and aquaculture is becoming a major source of dietary protein from aquatic products. Large yellow croaker ( Larimichthys crocea Large yellow croaker (Caucasus macrantha) plays a prominent role in aquaculture; therefore, growth-related traits are crucial for many aquatic species, directly impacting yield. However, challenges such as climate change, disease outbreaks, and loss of genetic diversity due to inbreeding hinder the growth and body shape of large yellow croaker. In most selective breeding programs, growth-related traits are considered quantitative traits controlled by multiple genes distributed throughout the genome. The body shape of large yellow croaker is vital to the aquaculture industry, influencing its swimming, feeding, and consumer preferences. Furthermore, since most consumers prefer slender large yellow croaker, body shape has become an important economic characteristic, significantly impacting commercial value. Given its importance, identifying the genes regulating the growth and body shape of large yellow croaker is essential.

[0003] Studies on model organisms have revealed that the transforming growth factor-β (TGFβ) family plays a crucial role in coordinating various biological processes such as cell growth, proliferation, and differentiation. The TGFβ signaling pathway involves ligand binding to membrane receptors, activation of type I receptors via phosphorylation of type II receptors, followed by phosphorylation of Smad proteins to initiate signal transduction. The TGFβ superfamily contains a wide variety of ligands, including TGFβs (transforming growth factor β, TGFB1-5), BMPs (bone morphogenetic proteins, BMP2-16), GDFs (growth and differentiation factors, GDF1-15), Nodal, activins (activating proteins, INHBA and INHBB), and repressor proteins. The receptors for these ligands are mainly divided into two categories: type I and type II. Specifically, type I receptors include ALK (activator-like kinases) 1 to 7, while type II receptors include TGFBR2, ACVR2, ACVR2B, AMHR2, and BMPR2. Smad proteins, which play key roles in downstream signaling pathways of these receptors, can be further divided into three distinct categories: receptor-activating or pathway-restricting Smads (R-Smads), co-pathway Smads (Co-Smads), and inhibitory Smads (I-Smads). This classification system provides a theoretical framework for in-depth exploration of the complex interaction mechanisms and signaling pathways within the TGFβ superfamily. Essentially, embryonic cells during development and various cell types in the adult body are capable of sensing TGFβ signals. Currently, studies have been conducted to assess the number and types of TGFβ family members in model organisms ranging from worms and flies to mammals. The vast majority (if not all) of cell types respond to at least a subset of TGFβ family ligands, thereby regulating numerous cellular activities.

[0004] The TGFβ signaling pathway plays a crucial role not only in mediating growth and development in organisms, but also, as revealed in recent genome-wide association studies of body shape-related traits in large yellow croaker, it is deeply involved in the regulation of these traits. However, to date, there is a lack of in-depth research exploring which specific members of this pathway are involved and how they are precisely regulated. This lack of research is mainly due to the fact that the whole-genome sequencing and detailed genetic mapping of large yellow croaker were not completed until 2014. In large yellow croaker, the TGFβ family includes 24 ligands, 19 receptors, and 3 Smads proteins. Furthermore, some members have more detailed subtype classifications, such as BMPR1BX1 (XM_010732488.3), BMPR1BX2 (XM_027275036.1), and BMPR1BX3 (XM_027275041.1). However, there are few studies that systematically classify these gene entries, and even fewer studies that delve into the specific mechanisms by which each member regulates the growth and body shape of the large yellow croaker. Summary of the Invention

[0005] To address the shortcomings of existing technologies, this invention provides a gene combination related to the body shape of large yellow croaker and its application in breeding. The provided gene combination can be used for screening research on large yellow croaker with a beautiful body shape, providing molecular markers for large yellow croaker breeding screening.

[0006] To solve the aforementioned technical problem, the technical solution adopted by the present invention is as follows:

[0007] This invention provides a gene combination related to the body shape of large yellow croaker, the gene combination including INHBB gene, Nodal gene, GDF3 gene, ACVR2A gene, ALK4 gene, and Smad2 gene.

[0008] In a preferred embodiment, the gene combination is INHBB-ACVR2A-ALK4-Smad2, Nodal-ACVR2A-ALK4-Smad2, and GDF3-ACVR2A-ALK4-Smad2.

[0009] The transforming growth factor β (TGFβ) signaling axis plays a crucial role in coordinating a range of biological functions, including cell growth, proliferation, and differentiation. This invention identifies the TGFβ signaling pathway members and their expression patterns in large yellow croaker under different farming conditions. The TGFβ signaling pathway and expression patterns in large yellow croaker raised under two different farming conditions were investigated: Group N (2,400 fish in a 120 cubic meter net cage) and Group V (168,000 fish in a 5,600 cubic meter aquaculture vessel). After 120 days, Group V fish showed faster growth and a more slender body compared to Group N. This invention employs bioinformatics techniques to systematically classify 48 TGFβ sequences from the National Center for Biotechnology Information (NCBI) database. Through rigorous data analysis and comparison, 21 TGFβ ligands, 10 TGFβ receptors, and 3 Smads proteins were accurately identified in large yellow croaker. To further explore the mechanisms of action of these signaling molecules, this invention further extracted RNA from large yellow croaker tissue samples containing the aforementioned identified components and used advanced molecular biology techniques to detect their mRNA expression levels. Experimental results showed that these signaling molecules mainly play a key regulatory role in the growth rate and body shape of large yellow croaker through three specific ligand-receptor-R-Smad signaling pathways: the INHBB-ACVR2A-ALK4-Smad2 signaling pathway; the Nodal-ACVR2A-ALK4-Smad2 signaling pathway; and the GDF3-ACVR2A-ALK4-Smad2 signaling pathway. Based on these innovative research findings, this invention successfully identified a set of gene combinations closely related to the regulation of large yellow croaker body shape, providing an important theoretical foundation and technical support for large yellow croaker breeding and related industries.

[0010] In a preferred embodiment, the amino acid sequence of the protein encoded by the INHBB gene is SEQ ID NO:1; the amino acid sequence of the protein encoded by the Nodal gene is SEQ ID NO:2; the amino acid sequence of the protein encoded by the GDF3 gene is SEQ ID NO:3; the amino acid sequence of the protein encoded by the ACVR2A gene is SEQ ID NO:4; the amino acid sequence of the protein encoded by the ALK4 gene is SEQ ID NO:5; and the amino acid sequence of the protein encoded by the Smad2 gene is SEQ ID NO:6.

[0011] In a preferred embodiment, the INHBB gene has a nucleic acid sequence encoding CDS of SEQ ID NO:7; the Nodal gene has a nucleic acid sequence encoding CDS of SEQ ID NO:8; the GDF3 gene has a nucleic acid sequence encoding CDS of SEQ ID NO:9; the ACVR2A gene has a nucleic acid sequence encoding CDS of SEQ ID NO:10; the ALK4 gene has a nucleic acid sequence encoding CDS of SEQ ID NO:11; and the Smad2 gene has a nucleic acid sequence encoding CDS of SEQ ID NO:12.

[0012] In another aspect, the present invention also provides the application of the genomic collaborations related to the body shape of large yellow croaker as described in any of the above technical solutions as molecular markers in screening large yellow croakers of different body shapes.

[0013] The present invention also provides a preparation for detecting the expression levels of each gene in the gene combination related to the body shape of large yellow croaker described in any of the above technical solutions, wherein the preparation is PCR amplification primers.

[0014] The present invention also provides a product for screening large yellow croaker of different body shapes. The product is a real-time PCR amplification detection kit. The product screens large yellow croaker of different body shapes by detecting the expression levels of each gene in the gene combination related to the body shape of large yellow croaker as described in any of the above technical solutions.

[0015] The present invention also provides a method for screening large yellow croakers of different body shapes. The method is to screen large yellow croaker individuals by detecting the expression level of genes in the gene combination described in any of the above technical solutions.

[0016] In a preferred embodiment, individuals with low expression levels of the INHBB gene, Nodal gene, GDF3 gene, ACVR2A gene, ALK4 gene, and Smad2 gene are screened.

[0017] The TGFβ signaling pathway binds to membrane receptors via ligands, activates type I receptors via type II receptor phosphorylation, and then phosphorylates Smad proteins, initiating signal transduction. The screening of gene combinations related to body shape in large yellow croaker was first based on the integrity of the aforementioned signaling pathways in the fish. Using the nucleotide and amino acid sequence integrity data of large yellow croaker from the NCBI database, we initially identified 15 signaling pathways. Then, based on the expression levels of ligands, receptors, and R-Smads in the experimental group, we further clarified the promoting and inhibiting effects of each gene. Furthermore, we identified feasible pathways for regulating body shape, comprising five pathways across two subfamilies. Based on the expression levels of each gene involved in the above five pathways in the muscles of group V, it was found that INHB, Nodal, GDF3, ACVR2A, ALK4, Smad2 and N group were all suppressed, further identifying three signaling pathways: INHB-ACVR2A-ALK4-Smad2, Nodal-ACVR2A-ALK4-Smad2 and GDF3-ACVR2A-ALK4-Smad2.

[0018] In a preferred embodiment, the method is performed by quantitative real-time PCR amplification; wherein, a primer pair for detecting the INHBB gene has primer sequences of SEQ ID NO:13 and SEQ ID NO:14; a primer pair for detecting the Nodal gene has primer sequences of SEQ ID NO:15 and SEQ ID NO:16; a primer pair for detecting the GDF3 gene has primer sequences of SEQ ID NO:17 and SEQ ID NO:18; a primer pair for detecting the ACVR2A gene has primer sequences of SEQ ID NO:19 and SEQ ID NO:20; a primer pair for detecting the ALK4 gene has primer sequences of SEQ ID NO:21 and SEQ ID NO:22; and a primer pair for detecting the Smad2 gene has primer sequences of SEQ ID NO:23 and SEQ ID NO:24.

[0019] Compared with the prior art, the beneficial effects of the present invention are as follows:

[0020] This invention analyzes the TGFβ signaling pathway and its expression patterns in large yellow croaker raised under different farming models. It finds that the INHBB-ACVR2A-ALK4-Smad2, Nodal-ACVR2A-ALK4-Smad2, and GDF3-ACVR2A-ALK4-Smad2 axes play a dominant role in regulating the growth and body shape of large yellow croaker. Furthermore, populations with low gene expression levels exhibit faster growth and slender body shapes, which can serve as molecular markers for screening large yellow croaker with rapid growth potential and slender body shapes. Attached Figure Description

[0021] Figure 1 The growth and body shape of the large yellow croaker provided in Embodiment 2 of the present invention, wherein A is the fish in the early stage of aquaculture, B is the fish cultured in net cages for 120 days, and C is the fish cultured in aquaculture boats for 120 days.

[0022] Figure 2 A phylogenetic tree was constructed based on the nucleotide sequences of ligands belonging to the TGFβ / activin / Nodal subfamily of large yellow croaker provided in Example 2 of this invention;

[0023] Figure 3 This is the phylogenetic tree constructed based on the nucleotide sequences of the BMP / GDF / MIS subfamily ligands of large yellow croaker provided in Example 2 of the present invention;

[0024] Figure 4 This is the phylogenetic tree constructed based on the receptor nucleotide sequence of large yellow croaker provided in Embodiment 2 of the present invention;

[0025] Figure 5 This is the phylogenetic tree constructed based on the R-Smads nucleotide sequence of large yellow croaker provided in Embodiment 2 of the present invention;

[0026] Figure 6 The expression of the ligand provided in Example 2 of this invention in the liver of large yellow croaker;

[0027] Figure 7 The expression of the ligand provided in Example 2 of this invention in the muscle of large yellow croaker;

[0028] Figure 8 The relative expression level of receptors in the liver of large yellow croaker provided in Example 2 of the present invention;

[0029] Figure 9 The relative expression of receptors in the muscle of large yellow croaker provided in Embodiment 2 of the present invention;

[0030] Figure 10 The relative expression of Smads in the liver provided in Embodiment 2 of the present invention;

[0031] Figure 11 This is the relative expression of Smads in muscle provided in Embodiment 2 of the present invention. Detailed Implementation

[0032] Large yellow croaker (Group C) with an average total length of (32.41±1.16) cm and an average weight of (350.4±34.24) g were selected and divided into two experimental groups. Group N consisted of 2,400 fish, which were raised in a 120 cubic meter net cage; Group V consisted of 168,000 fish, which were raised in the hold of a 5,600 cubic meter aquaculture vessel. After 120 days of feeding, 15 fish were randomly selected from each group for further detailed experimental analysis. The study used parameters such as body weight, total length, body length, body depth, chest height, caudal peduncle length, caudal peduncle width, caudal peduncle length to caudal peduncle width ratio (CPL / CPW), body length to body depth ratio (BL / BD), caudal peduncle length to body length ratio (CPL / BL), and condition factor to describe growth and body shape.

[0033] The present invention will now be described in detail with reference to specific embodiments and accompanying drawings.

[0034] Example 1: Sequence analysis of genes related to growth and body shape in large yellow croaker

[0035] This invention first downloads the complete protein sequences of TGFβ ligands, receptors, and Smads from the National Center for Biotechnology Information (NCBI) database of large yellow croaker, and uses BLAST software to compare and analyze them with corresponding sequences of other species. Then, multiple sequence alignment is performed using the ClustalW multiple sequence alignment tool (http: / / www.ebi.ac.uk / clustalw / ), and amino acid sequences of closely related ligands, receptors, and Smads are screened and downloaded based on the alignment results. Subsequently, a phylogenetic tree of individuals is constructed using the neighbor-joining (NJ) method of MEGA 7.0 software, and the robustness of the phylogenetic tree is evaluated through 1000 guided replicates to ensure its reliability.

[0036] Table 1. Oligonucleotide primers used for amplifying the TGFβ signaling pathway genes in large yellow croaker.

[0037]

[0038]

[0039]

[0040] Statistical analysis was performed on all data using GraphPad Prism 5.0 software, and one-way ANOVA was used for evaluation. Tukey's Honestly Significant Difference (HSD) test was used for post-hoc multiple comparisons to assess differences between means. Results are expressed as mean ± SD, and a p-value less than 0.05 was considered statistically significant.

[0041] Example 2: Differentially expressed genes in groups N and V

[0042] (1) Growth and body shape of large yellow croaker under two farming models

[0043] After a 120-day rearing period, significant differences were observed between group V and group N in terms of body weight, total length, body length, caudal peduncle length, caudal peduncle length / width, body length / body depth, caudal peduncle length / body length, and condition factor. Group V grew faster and had a more slender body shape than group N (Table 2). To more intuitively represent these differences, one fish from each of the three groups was selected as a representative. Figure 1 ).

[0044] Table 2. Effects of cage and boat aquaculture on growth and body shape of large yellow croaker (n=15)

[0045]

[0046] (2) Phylogenetic analysis of ligand sequences

[0047] The types and quantities of ligands, such as Figure 2 and Figure 3 As shown. NCBI contains 24 ligand entries, which can be divided into 21 classes. It is noteworthy that GDF15 in large yellow croaker (XM_010733382.3) does not cluster with other GDF species in the phylogenetic tree, but is more closely related to GDF8. Figure 3 Except for AMH / MIS, which has four subtypes, the other 20 ligands each have only one subtype. In large yellow croaker, the identified ligand types include GDF8, GDF6 / BMP13, GDF5 / BMP14, BMP3, GDF9, AMH, BMP10, BMP4 / BMP2B, BMP2 / BMP2A, GDF3, BMP8 / BMP8B, BMP5, BMP7, BMP6, NDR2, INHBB / Activin B, INHBA / Activin A, TGFB1, TGFB2, and TGFB3.

[0048] (3) Phylogenetic analysis of receptors

[0049] Through research Figure 4The receptor phylogenetic tree in the database reveals that the 19 receptor nucleotide sequence entries in the gene pool can be divided into 10 different groups. These include five type I receptors: ALk3 / BMPR1A, ALK6 / BMPR1B, ALK2 / ACVR1, ALK4 / ACVR1B, and ALK7 / ACVR1C. There are also five type II receptors: ACVR2A / ACTRⅡA, ACVR2B / ACTRⅡB, TGFBR2 / TBRⅡ, BMPR2, and AMHR2 / MISR2. Some of these types exhibit different subtypes. For example, AMHR2 has two subtypes, AMHR2X1 and AMHR2X2; ACVR2A has five subtypes, ACVR2AX1, ACVR2AX2, ACVR2AX3, ACVR2AX4 and ACVR2AX5; BMPR1A has two subtypes, BMPR1AX1 and BMPR1AX2; and BMPR1B has four subtypes, BMPR1BX1, BMPR1BX2, BMPR1BX3 and BMPR1BX4.

[0050] (4) Phylogenetic analysis of R-Smads

[0051] exist Figure 5 In the phylogenetic tree shown, the nucleotide sequences of each type of R-Smad are grouped together. The five Smad genes found in large yellow croaker are classified into three classes: Smad1, Smad2, and Smad3. Among them, Smad2 includes three different subtypes: Smad2X1, Smad2X2, and Smad2X3. In large yellow croaker, although Smad5 and Smad8 belong to the R-Smads family and primarily respond to BMP signaling, they were not identified.

[0052] (5) mRNA level of ligands

[0053] In this invention study, the expression of the ligand in the liver tissue of large yellow croaker is shown in [reference needed]. Figure 6 Among them, TGFB2, BMP6, and GDF9 showed significantly upregulated expression levels in both groups N and V; TGFB1, BMP3, BMP4, BMP10, and GDF5 showed decreased expression levels in group N and increased expression levels in group V; BMP5, BMP7, and GDF6 showed no significant difference in expression levels between groups N and V; in muscle tissue ( Figure 7 The expression patterns of ligands in muscle tissue are quite different from those in the liver, except for GDF9. Furthermore, the expression levels of BMP4, BMP5, and AMH were significantly increased in both the N and V groups. These findings suggest that ligand expression in muscle tissue may be influenced by regulatory mechanisms different from those in the liver.

[0054] (6) mRNA level of receptor

[0055] In the research of this invention, Figure 8 The expression levels of ligands in the liver tissue of large yellow croaker were shown. There was no significant difference in the expression levels of ALK2 and AMHR2 between groups N and V, while the expression levels of ALK3 and ALK7 decreased in group N and increased in group V. Notably, except for ALK4, the mRNA expression trends of the three receptors ALK6, TGFBR2, and ACVR2B in muscle tissue differed from those in liver tissue. Figure 8 Furthermore, in muscle tissue, there was no significant difference in ALK6 expression levels between groups N and V, while the expression of ALK7, TGFBR2, and ACVR2B was significantly increased in both groups N and V. Figure 9 ).

[0056] (7) mRNA level of R-Smads

[0057] In the study of this invention, Figure 10 and Figure 11 The expression patterns of R-Smads in the liver and muscle tissues of large yellow croaker were presented separately. Specifically, in both liver and muscle tissues, there were no significant changes in Smad3 expression levels between the experimental and control groups, and no significant difference in Smad3 expression between the two experimental groups. Regarding Smad1 and Smad2, their expression patterns in the liver and muscle tissues of group N showed a differentiated trend. In liver tissue, the expression of these two genes differed from the control group, while no similar differences were found in muscle tissue; conversely, genes that showed significant differences in muscle tissue compared to the control group did not show differences in liver tissue. This result indicates that the expression of Smad1 and Smad2 in group N is affected by tissue-specific regulatory mechanisms. However, in group 5, the expression of Smad1 and Smad2 did not appear to be affected by the tissue type used in the experiment; that is, the expression patterns of the two genes remained consistent in both tissues, and no inter-tissue differences were observed. In summary, under different treatment methods and tissue types, the expression of Smad3 in R-Smads of large yellow croaker is relatively stable, while the expression of Smad1 and Smad2 shows tissue-specific variation patterns under different treatment conditions.

[0058] Analysis results

[0059] Compared to the slender body of wild large yellow croaker, farmed large yellow croaker often has a more rounded appearance. The elongated shape of these fish demonstrates superior swimming, foraging, and predator avoidance abilities. Consumer preference for slender, elongated large yellow croaker significantly influences its market value. In a 120-day farming experiment, large yellow croaker farmed on a workboat grew faster and had a more elongated body compared to those farmed in net cages. The observed faster growth rate is likely attributed to the workboat's advanced water exchange system, which enables continuous forced water exchange between the farming tanks and the external natural seawater. Furthermore, the workboat utilizes a deep-water intake device to obtain seawater with the optimal temperature and salinity required for farming, ensuring the large yellow croaker maintains optimal growth conditions. It is inferred that the swimming behavior and schooling tendency of large yellow croaker farmed in the swimming-mode workboat farming system are very similar to those of wild large yellow croaker, which is the main reason for their faster growth rate and more elongated body shape.

[0060] Based on extensive research into the specificity of TGFβ superfamily ligand-receptor-R-Smad interactions, and experience in tracing the TGFβ signaling pathway in sea cucumber and Pacific oyster, relevant nucleotide sequences in the large yellow croaker genome were detected. Figure 2-5 ) and their mRNA expression levels ( Figure 6-11 Table 3 summarizes the ligand-receptor-R-Smad correspondences in large yellow croaker. In the TGFβ / activin / Nodal subfamily, detected components included ligands (INHBA, INHBB, and Nodal), receptor I (ALK4 and ALK7), receptor II (ACVR2A and ACVR2B), and R-Smads (Smad2, 3). Conversely, in the BMP / GDF / AMH subfamily, a series of ligands (BMP2, BMP4, BMP5, BMP6, BMP7, BMP8A, GDF3, GDF5, GDF6, GDF8, GDF9, and AMH), receptor I (ALK2, ALK3, ALK4, ALK6, and ALK7), receptor II (ACVR2A, ACVR2B, BMPR2, and AMHR2), and R-Smads (Smad1, 2, 3) were detected. Table 3 lists family members exhibiting complete signaling pathways. Some members, including TGFB1-3, possess nucleotide and amino acid sequences in the large yellow croaker genome, and their mRNA expression levels under different culture conditions were assessed using RT-PCR. However, after reviewing currently reported TGFB1-3 receptor types, we did not find these genes in the large yellow croaker genome. Therefore, these members are not included in Table 3 due to their incomplete signaling pathways.

[0061] Table 3. Specificity of TGFβ superfamily ligand-receptor-R-Smad

[0062]

[0063]

[0064] Based on the promoting and inhibiting effects observed in the experimental group by ligands, receptors, and R-Smads ( Figure 6-11 Table 4 lists feasible pathways regulating liver growth and body shape, including 11 pathways across two subfamilies. Table 5 records feasible pathways regulating muscle growth and body shape, with 5 pathways across two subfamilies. Fewer feasible pathways exist in muscle compared to the liver, likely because different culture methods do not activate Smad1. Notably, Smad1 is the only recognizable R-Smad among various BMPs in large yellow croaker. This protein can bind to Co-Smad (Smad4) to form a complex, which then enters the nucleus to regulate gene expression, as mentioned in related studies. The five pathways regulating growth and body shape are also present in the liver of large yellow croaker muscle tissue. However, whether these pathways maintain consistent tissue stability in different organs (such as the heart, spleen, kidneys, intestines, gonads, and brain) remains unanswered and requires further experimental verification in these other tissues. In group V muscle, the expression levels of each member in the three signaling pathways were different: the expression levels of each member in the INHBB-ACVR2A-ALK4-Smad2, Nodal-ACVR2A-ALK4-Smad2, and GDF3-ACVR2A-ALK4-Smad2 signaling pathways were inhibited. Figure 7 , 9 (11). INHBB, Nodal, and GDF3, as ligands, bind to specific receptors to transmit signals; reduced expression of these ligands may lead to weakened signaling capabilities. ACVR2A and ALK4 can bind to these three ligands, thereby activating downstream signal transduction of Smad2. Reduced expression of these ligands may further inhibit TGFβ signaling. As a downstream signaling molecule, reduced Smad2 expression indicates decreased activity of the TGFβ signaling pathway. Although ACVR2B and ALK7 can also bind to the above three ligands, their expression levels are increased. They may bind to other ligands to transmit different signals or compensate for reduced function of ALK4 and ACVR2A under specific conditions.

[0065] Table 4. TGFβ superfamily ligand-receptor-R-Smad specificity in the liver

[0066]

[0067] Table 5. TGFβ superfamily ligand-receptor-R-Smad specificity in muscle

[0068]

[0069] In summary, this invention discloses the large yellow croaker ( Larimichthys crocea The large yellow croaker contains 34 members belonging to two TGFβ subfamilies. These signaling molecules may affect growth rate and body shape through five different ligand-receptor-R-Smad signaling pathways. Among them, the INHBB-ACVR2A-ALK4-Smad2, Nodal-ACVR2A-ALK4-Smad2, and GDF3-ACVR2A-ALK4-Smad2 signaling axes play a dominant role in regulating the body shape of the large yellow croaker.

Claims

1. The application of a gene combination in screening large yellow croakers of different body shapes, characterized in that, The gene combination mentioned includes the INHBB gene, Nodal gene, GDF3 gene, ACVR2A gene, ALK4 gene, and Smad2 gene; The gene combination mentioned is INHBB-ACVR2A-ALK4-Smad2, Nodal-ACVR2A-ALK4-Smad2, or GDF3-ACVR2A-ALK4-Smad2; The amino acid sequence of the protein encoded by the INHBB gene is SEQ ID NO:1; the amino acid sequence of the protein encoded by the Nodal gene is SEQ ID NO:2; the amino acid sequence of the protein encoded by the GDF3 gene is SEQ ID NO:3; the amino acid sequence of the protein encoded by the ACVR2A gene is SEQ ID NO:4; the amino acid sequence of the protein encoded by the ALK4 gene is SEQ ID NO:5; and the amino acid sequence of the protein encoded by the Smad2 gene is SEQ ID NO:

6.

2. The application according to claim 1, characterized in that, The INHBB gene, whose nucleic acid sequence encoding CDS is SEQ ID NO:7; the Nodal gene, whose nucleic acid sequence encoding CDS is SEQ ID NO:8; the GDF3 gene, whose nucleic acid sequence encoding CDS is SEQ ID NO:9; the ACVR2A gene, whose nucleic acid sequence encoding CDS is SEQ ID NO:10; the ALK4 gene, whose nucleic acid sequence encoding CDS is SEQ ID NO:11; and the Smad2 gene, whose nucleic acid sequence encoding CDS is SEQ ID NO:12.

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