A method for strengthening yellow croaker parent

Abstract

The present invention provides a yellow croaker parent strengthening method, comprising the following steps: S1, selecting healthy yellow croaker with clear genetic background as a parent; S2, performing transcriptome sequencing on the selected parent to analyze its sex reversal process and molecular mechanism; S3, screening differentially expressed genes related to sex development according to the transcriptome sequencing results; S4, using molecular biology technology to perform functional verification on the screened key genes for sex development; S5, according to the functional verification results, selecting yellow croaker with excellent traits as a parent for breeding; S6, performing genetic background analysis on the offspring after breeding to ensure genetic stability and the transmission of excellent traits; S7, during the breeding process, monitoring the composition of water microbial communities and intestinal flora of yellow croaker to ensure the health of the breeding environment; S8, adjusting the breeding conditions according to the monitoring results, optimizing the breeding environment, so as to promote the health and reproductive capacity of the parents. The growth rate, reproductive capacity and environmental adaptability of yellow croaker are improved.

Description

Technical Field

[0001] The invention relates to the technical field of aquaculture, in particular to a method for strengthening yellow croaker parents. Background Art

[0002] In the field of aquaculture, as an important economic fish, the development of the aquaculture industry of yellow croaker has received extensive attention. The quality of yellow croaker parents is directly related to the growth rate, disease resistance and overall yield of the offspring. Therefore, how to improve the quality of yellow croaker parents and enhance their reproductive capacity is a problem that aquaculture operators and scientific researchers have long been committed to solving.

[0003] Traditional methods of strengthening yellow croaker parents mainly include improving the breeding environment, optimizing feed formula, controlling breeding density, etc. However, these methods often have certain limitations, such as the difficulty in accurately controlling environmental improvement, the lack of targeted feed optimization, and the adjustment of breeding density may affect the health of the parents. In addition, with the expansion of breeding scale and the increase in market demand, traditional methods can no longer meet the needs of modern aquaculture for high efficiency, high yield and high disease resistance.

[0004] However, there are certain limitations in its growth rate, reproductive capacity and adaptability to the environment, which restricts the further development of the yellow croaker farming industry. The growth and reproductive performance of yellow croaker are closely related to the composition of its intestinal microbial community. By analyzing the changes in the composition of the microbial community in the aquaculture water and the intestinal flora of yellow croaker, it is possible to have a potential impact on the feeding and growth processes of yellow croaker. In addition, the sexual development and gonadal development process of yellow croaker plays a decisive role in its reproductive performance, but the molecular mechanism of this process is currently limited.

[0005] Therefore, developing a method that can effectively improve the quality of yellow croaker parents and enhance their growth and reproductive capacity is of great significance to promoting the development of yellow croaker breeding industry. Summary of the invention

[0006] The purpose of the present invention is to provide a method for strengthening yellow croaker parents to solve the problems in the prior art.

[0007] To achieve the above object, the present invention provides the following technical solution: The present invention provides a method for strengthening the parent of yellow croaker, comprising the following steps:

[0008] S1. Select healthy yellow croaker with clear genetic background as parents;

[0009] S2. Transcriptome sequencing of the selected parents to analyze the sex reversal process and molecular mechanism;

[0010] S3. Based on the transcriptome sequencing results, differentially expressed genes related to sex development were screened;

[0011] S4. Use molecular biology techniques to verify the functions of the key genes for sex development that have been screened;

[0012] S5. According to the functional verification results, yellow croaker with excellent traits is selected as the parent for breeding;

[0013] S6. Conduct genetic background analysis on the offspring after reproduction to ensure genetic stability and the transmission of excellent traits;

[0014] S7. During the breeding process, monitor the composition of water microbial flora and the intestinal flora of yellow croaker to ensure the health of the breeding environment;

[0015] S8. Adjust breeding conditions and optimize the breeding environment based on monitoring results to promote the health and reproductive capacity of the parents.

[0016] Preferably, in step S2, transcriptome sequencing selects three morphological gonads (mature testis, facultative male and female gonads, and mature ovaries) of yellow croaker for sequencing analysis.

[0017] Preferably, in step S3, differentially expressed genes related to sex development are screened by comparative transcriptome analysis, and the genes include sox3 / 9a / 9b / 10, dmrt1 / 2 / 3 / 4 / 5, piwil1 / 2, foxl2 / h1, dax1, zp3, sycp1 / 2 / 3, and odf2 / 3.

[0018] Preferably, in step S7, the water microbial community and the intestinal flora of yellow croaker are monitored to develop 110 OTU markers of water microbial community and 34 OTU markers of intestinal flora of yellow croaker.

[0019] Preferably, in step S8, the aquaculture environment is optimized and the salinity, temperature and pH parameters of the aquaculture water are adjusted to simulate the environmental conditions for the natural growth of yellow croaker.

[0020] Preferably, the parental enhancement method further comprises tracking and observing the whole process of gonadal development of the offspring to ensure the normality of gonadal development and the correct transmission of parental genetic information.

[0021] Preferably, the parental enhancement method further comprises performing comparative transcriptome analysis on the offspring to screen out differentially expressed genes associated with sex development and verify their functions.

[0022] Preferably, the parental enhancement method further comprises tracking and observing the timeline of gonadal development of the offspring to clarify the important time nodes in the gonadal development process.

[0023] Preferably, the parental enhancement method further comprises observing the gonadal development structure of the offspring to ensure the normality of gonadal development and the correct transmission of parental genetic information.

[0024] Preferably, the parental enhancement method further comprises performing genetic background analysis on the offspring to ensure genetic stability and the transmission of excellent traits.

[0025] The present invention has at least the following beneficial effects:

[0026] The present invention proposes a method for strengthening yellow croaker parents. By developing OTU markers of aquaculture water microbial communities and yellow croaker intestinal flora, the aquaculture environment can be effectively monitored and regulated, thereby optimizing the growth environment of yellow croaker. By screening key genes for sex development of yellow croaker through comparative transcriptome analysis, the sex reversal process and molecular mechanism of yellow croaker can be deeply understood, providing a theoretical basis for improving sex control and reproductive performance. In addition, by tracking the entire process of gonadal development of yellow croaker, the important time nodes in the gonadal development process can be clarified, providing a scientific basis for the selection and strengthening of yellow croaker parents. The present invention can improve the growth rate, reproductive capacity and adaptability of yellow croaker to the environment, thereby promoting the sustainable development of the yellow croaker breeding industry, and has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0027] Figure 1 A flow chart of the method provided by the present invention;

[0028] Figure 2 A graph showing the ATPase activity levels in the intestinal tissue, liver tissue and serum of yellow croaker provided by the present invention;

[0029] Figure 3 A graph showing relevant antioxidant indexes in the intestinal tissue and liver tissue of yellow croaker provided by the present invention;

[0030] Figure 4 The composition analysis diagram of the microbial community of aquaculture water and the intestinal flora of yellow croaker provided by the present invention;

[0031] Figure 5 A development diagram of markers for the intestinal flora of yellow croaker provided by the present invention;

[0032] Figure 6 A development map of markers for the microbial community of aquaculture water provided by the present invention;

[0033] Figure 7 The functional gene differential expression analysis diagram provided by the present invention;

[0034] Figure 8 A timeline diagram of gonadal development of yellow croaker provided by the present invention;

[0035] Fig. 9The present invention provides a diagram of the gonad structure of the yellow croaker at different developmental stages. DETAILED DESCRIPTION

[0036] Unless otherwise defined, all technical and scientific terms used in this specification have the same meanings as those commonly understood by technicians in the technical field to which the invention belongs. The terms used in this specification and in the description of the present invention are only for the purpose of describing specific embodiments and are not used to limit the present invention. The term "and / or" used in this specification includes any and all combinations of one or more related listed items.

[0037] Example

[0038] like Figure 1-9 As shown, a method for strengthening yellow croaker parents may include the following steps:

[0039] S1: Select healthy yellow croaker as the research subject and ensure that it has a good genetic background and growth status.

[0040] S2: The gonadal development of yellow croaker was morphologically observed, and the various stages of gonadal development were recorded. According to the morphological characteristics of the gonads, yellow croaker was divided into three groups: mature testes, facultative hermaphroditic gonads, and mature ovaries.

[0041] S3: Using transcriptome sequencing technology, the gonadal tissues of the above three groups of yellow croaker were sequenced and analyzed to obtain transcriptome data.

[0042] S4: Through comparative transcriptome analysis, differentially expressed genes related to sex development were screened out, including sox3 / 9a / 9b / 10, dmrt1 / 2 / 3 / 4 / 5, piwil1 / 2, foxl2 / h1, dax1, zp3, sycp1 / 2 / 3, odf2 / 3, etc.

[0043] Specifically, Figure 7 (A) Volcano plot of differentially expressed genes between testis and facultative gonads; (B) Volcano plot of differentially expressed genes between facultative gonads and ovaries; (C) Volcano plot of differentially expressed genes between testis and ovaries.

[0044] S5: Use molecular biology techniques, such as CRISPR / Cas9 gene editing technology, to functionally verify the key genes for sex development that have been screened out to determine their specific role in sex development.

[0045] S6: Based on the functional verification results, select yellow croaker with excellent traits as parents for breeding to ensure its genetic stability and the transmission of excellent traits.

[0046] S7: Conduct genetic background analysis on the offspring after reproduction, including genomic DNA extraction, PCR amplification, sequencing and other steps to ensure genetic stability and the transmission of excellent traits.

[0047] S8: During the breeding process, monitor the composition of water microbial communities and the intestinal flora of yellow croaker, and develop 110 OTU markers of water microbial communities and 34 OTU markers of intestinal flora of yellow croaker to ensure the health of the breeding environment.

[0048] S9: According to the monitoring results, adjust the breeding conditions and optimize the breeding environment, such as adjusting the salinity, temperature, and pH parameters of the breeding water to simulate the environmental conditions for the natural growth of yellow croaker.

[0049] S10: Track and observe the entire process of gonadal development of the offspring, including morphological observation of gonadal tissue, tracking observation of the gonadal development timeline, and observation of gonadal development structure, to ensure the normal development of gonads and the correct transmission of parental genetic information.

[0050] Specifically, Figure 8 , Fig. 9 Middle, MO, mature oocyte; PO, primary oocyte; PSC, primary spermatocyte; SC, spermatocyte; SG, spermatogonia; SP, spermatid; SZ, spermatozoon;

[0051] (AD) 1st age; (EH) 2nd age; (IK) 3rd age. MO, mature oocyte; OT, ovarian tissue; PO, primary oocyte

[0052] oocyte; PVO, previtellogenic oocyte; SC, spermatocyte; SG, spermatogonia; SP, spermatid; SZ, sperm; TT, testicular tissue; VO, midvitellogenic oocyte.

[0053] Through the above embodiments, the present invention provides a method for strengthening the parent of yellow croaker, which can effectively improve the growth rate, reproductive capacity and environmental adaptability of yellow croaker, and promote the sustainable development of the yellow croaker breeding industry.

[0054] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit them. Under the concept of the present invention, the technical features in the above embodiments or different embodiments may also be combined, the steps may be implemented in any order, and there are many other changes in different aspects of the present invention as above, which are not provided in detail for the sake of simplicity. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they may still modify the technical solutions described in the aforementioned embodiments, or make equivalent replacements for some of the technical features therein. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for strengthening yellow croaker parent, characterized in that: The following steps are involved: S1. Select healthy yellow croaker with clear genetic background as parents; S2. Transcriptome sequencing of the selected parents to analyze the sex reversal process and molecular mechanism; S3. Based on the transcriptome sequencing results, differentially expressed genes related to sex development were screened; S4. Use molecular biology techniques to verify the functions of the key genes for sex development that have been screened; S5. According to the functional verification results, yellow croaker with excellent traits is selected as the parent for breeding; S6. Conduct genetic background analysis on the offspring after reproduction to ensure genetic stability and the transmission of excellent traits; S7. During the breeding process, monitor the composition of water microbial flora and the intestinal flora of yellow croaker to ensure the health of the breeding environment; S8. Adjust breeding conditions and optimize the breeding environment based on monitoring results to promote the health and reproductive capacity of the parents.

2. The method for strengthening yellow croaker parent according to claim 1, characterized in that: In step S2, transcriptome sequencing selects three morphological gonads (mature testis, facultative male and female gonads, and mature ovaries) of yellow croaker for sequencing analysis.

3. The method for strengthening yellow croaker parent according to claim 1, characterized in that: In step S3, differentially expressed genes related to sex development are screened by comparative transcriptome analysis, and the genes include sox3 / 9a / 9b / 10, dmrt1 / 2 / 3 / 4 / 5, piwil1 / 2, foxl2 / h1, dax1, zp3, sycp1 / 2 / 3, and odf2 / 3.

4. The method for strengthening yellow croaker parent according to claim 1, characterized in that: In step S7, water microbial communities and yellow croaker intestinal flora are monitored to develop 110 water microbial community OTU markers and 34 yellow croaker intestinal flora OTU markers.

5. The method for strengthening yellow croaker parent according to claim 1, characterized in that: In step S8, the aquaculture environment is optimized to adjust the salinity, temperature and pH parameters of the aquaculture water body to simulate the environmental conditions for the natural growth of yellow croaker.

6. The method for strengthening yellow croaker parent according to claim 1, characterized in that: The parental strengthening method also includes tracking and observing the whole process of gonadal development of offspring to ensure the normality of gonadal development and the correct transmission of parental genetic information.

7. The method for strengthening yellow croaker parent according to claim 1, characterized in that: The parental enhancement method also includes performing comparative transcriptome analysis on the offspring to screen out differentially expressed genes related to sex development and verify their functions.

8. The method for strengthening yellow croaker parent according to claim 1, characterized in that: The parental enhancement method also includes tracking and observing the timeline of gonad development of offspring to clarify important time nodes in the gonad development process.

9. The method for strengthening yellow croaker parent according to claim 1, characterized in that: The parental enhancement method also includes observing the gonad development structure of the offspring to ensure the normality of gonad development and the correct transmission of parental genetic information.

10. The method for strengthening yellow croaker parent according to claim 1, characterized in that: The parental enhancement method also includes performing genetic background analysis on the offspring to ensure genetic stability and the transmission of excellent traits.

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