Breeding method for high-temperature-resistant new species of sal-family large yellow croaker

Abstract

The invention relates to the field of fish breeding, and discloses a breeding method of a high-temperature-resistant new species of Halitium larimichthys crocea body, which comprises the following steps: performing sample collection on cultured Halitium larimichthys crocea, and screening high-temperature-resistant samples based on parameters such as survival rate and heat stress physiological indexes; the genetic potential of the high-temperature-resistant sample is further evaluated in a parent genetic evaluation mode, and the purpose of screening a core breeding sample is achieved. Finally, seedling raising is carried out by designing a low-inbreeding pairing scheme, heat-resistant advantage related genes are polymerized, and the purpose of fixing high-temperature-resistant characters is achieved through continuous breeding. The method can break through the bottleneck that the sal-family pseudosciaena crocea dies at a high temperature, realizes cultivation of a new high-temperature-resistant variety of the sal-family pseudosciaena crocea, and provides sustainable provenance support for expanding a pseudosciaena crocea culture space and a marine fish culture industry under a warming background.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of fish breeding, in particular, a high-temperature-resistant new variety breeding method of Pseudosciaena polyactis. BACKGROUND

[0002] Pseudosciaena polyactis belongs to Perciformes, Sciaenidae, Pseudosciaena, and is commonly known as yellow croaker, yellow cucumber fish, yellow flower fish, etc. It is a warm-temperate fish of the North Pacific region, with an adaptive range of water temperature of 10-32℃, and an optimal growth temperature of 18-25℃. It mainly distributes in the southern Yellow Sea, the East China Sea, and the coastal waters of Leizhou Peninsula in Guangdong. According to the 2025 China Fishery Statistical Yearbook, the 2024 domestic yellow croaker aquaculture yield was 292,600 tons, and the aquaculture area was concentrated in Zhejiang and Fujian. Due to the temperature limitation of the yellow croaker population, the 2024 Guangdong yellow croaker aquaculture yield was only 682 tons, mainly concentrated in Nan'ao Island and Shantou, etc.

[0003] At present, due to the severe challenge of global warming, the temperature range of yellow croaker is also relatively narrow, resulting in reduced feeding, decreased immunity, and increased mortality of yellow croaker. High temperature leads to growth stagnation, disease outbreak, and forced extension of the breeding cycle, resulting in increased feed costs. Pseudosciaena polyactis of the Pseudosciaena polyactis species is a characteristic yellow croaker population in the South China Sea, which has potential high-temperature resistance and is expected to develop into a suitable variety for tropical aquaculture. However, with the decreasing wild resources of Pseudosciaena polyactis, and the unclear genetic background and lagging genetic improvement of Pseudosciaena polyactis, the protection and development and utilization of Pseudosciaena polyactis genetic resources are severely restricted. Therefore, developing a high-temperature-resistant new variety of Pseudosciaena polyactis and cultivating a new variety of Pseudosciaena polyactis suitable for aquaculture in the South China Sea can effectively expand the aquaculture area of Pseudosciaena polyactis, thereby promoting the healthy and sustainable development of the Pseudosciaena polyactis aquaculture industry. In view of this, a high-temperature-resistant new variety breeding method of Pseudosciaena polyactis is proposed. SUMMARY

[0004] The present application overcomes the shortcomings of the prior art and provides a high-temperature-resistant new variety breeding method of Pseudosciaena polyactis.

[0005] To achieve the above-mentioned purposes, the technical scheme adopted by the present application is as follows:

[0006] The present application provides a high-temperature-resistant new variety breeding method of Pseudosciaena polyactis in the first aspect, comprising the following steps:

[0007] Sample collection is performed on Pseudosciaena polyactis to achieve the purpose of screening high-temperature-resistant samples;

[0008] The parent genetic evaluation analysis is performed on the high-temperature-resistant samples, genetic potential of the high-temperature-resistant samples is calculated, and the core breeding samples are screened from the high-temperature-resistant samples;

[0009] The core breeding samples are hybridized, heat-resistant related genes in the core breeding samples are aggregated, and generation selection is performed, so that the high-temperature-resistant trait of the new Pseudosciaena polyactis variety of the Lutao group is fixed.

[0010] Further, in a preferred embodiment of the present application, the high-temperature-resistant samples are screened by collecting samples of the Lutao group of Pseudosciaena polyactis and performing high-temperature stress experiments after the sample collection, and the specific process is as follows:

[0011] The samples of the Lutao group of Pseudosciaena polyactis are collected in a Lutao group of Pseudosciaena polyactis farm, and the collected samples of the Lutao group of Pseudosciaena polyactis cover different families, wherein the family is the Lutao group of Pseudosciaena polyactis samples with mating behavior and parentage index, and the samples of the Lutao group of Pseudosciaena polyactis are marked as target samples;

[0012] The phenotypic information of all the collected target samples is recorded, including the age, size and health status of the target samples;

[0013] All the target samples are randomly grouped, and different high-temperature breeding temperatures are set for different groups of target samples to perform gradient high-temperature stress experiments, and a control group is set, wherein the breeding temperature of the target samples in the control group is the standard breeding temperature of the target samples;

[0014] During the gradient high-temperature stress experiment, the core indexes of different groups of target samples are recorded in real time, wherein the core indexes include the survival rate, heat-resistant time and physiological and biochemical indexes of the target samples;

[0015] The heat-resistant time is the time from the beginning of stress to death of the target samples, or the time from the beginning of stress to the appearance of heat stress symptoms;

[0016] Among different groups of target samples, the target samples with the highest survival rate and the longest heat-resistant time are screened from different groups of target samples according to the corresponding core indexes, and are marked as high-temperature-resistant samples.

[0017] Further, in a preferred embodiment of the present application, the parent genetic evaluation analysis is performed on the high-temperature-resistant samples, genetic potential of the high-temperature-resistant samples is calculated, and the core breeding samples are screened from the high-temperature-resistant samples, and the specific process is as follows:

[0018] Subcutaneous electronic tags are implanted in all the high-temperature-resistant samples, and the tag information recorded in the implanted electronic tags is the phenotypic information of the corresponding high-temperature-resistant samples and the implantation date;

[0019] The implanted electronic tags can be used to locate the high-temperature-resistant samples during mating of different high-temperature-resistant samples.

[0020] The tail fin tissues of all the high-temperature-resistant samples are cut, and the DNA of the cut tail fin tissues of the high-temperature-resistant samples is extracted by using a DNA kit to obtain high-temperature-resistant sample DNA.

[0021] The high-temperature-resistant sample DNA is subjected to PCR amplification, and a big data network is introduced to analyze the target gene effect of the PCR-amplified high-temperature-resistant samples, determine the SNP sites related to the high-temperature-resistant trait, and mark the SNP sites related to the high-temperature-resistant trait as SNP sites related to the high-temperature-resistant trait.

[0022] The high-temperature-resistant samples are subjected to genotyping, which is to calculate the number of SNP sites related to the high-temperature-resistant trait in the high-temperature-resistant samples, sort all the high-temperature-resistant samples in reverse order according to the number of SNP sites related to the high-temperature-resistant trait, and calculate the homozygosity of the SNP sites related to the high-temperature-resistant trait in different high-temperature-resistant samples, and then sort the high-temperature-resistant samples again according to the homozygosity to output a preliminary genetic potential ranking table of the high-temperature-resistant samples.

[0023] The high-temperature-resistant samples ranked in the preset top are screened and marked as high-temperature-resistant samples of the first category in combination with the preliminary genetic potential ranking table of the high-temperature-resistant samples.

[0024] The genetic parameters of all the high-temperature-resistant samples of the first category are evaluated in combination with the tag information and the corresponding SNP sites related to the high-temperature-resistant trait, and the core breeding samples are screened according to the evaluation results.

[0025] Further, in a preferred embodiment of the present application, the genetic parameters of all the high-temperature-resistant samples of the first category are evaluated in combination with the tag information and the corresponding SNP sites related to the high-temperature-resistant trait, and the core breeding samples are screened according to the evaluation results, specifically:

[0026] The tag information and the corresponding SNP sites related to the high-temperature-resistant trait of the high-temperature-resistant samples of the first category are introduced into the animal model analysis software, and the genetic force of all the high-temperature-resistant samples of the first category is calculated by using the animal model analysis software.

[0027] The genetic force calculation is to combine the high-temperature-resistant samples of the first category freely and construct a kinship matrix, and the number of SNP sites related to the high-temperature-resistant trait and the homozygosity of the fry obtained by further mating of different combinations of the high-temperature-resistant samples of the first category are predicted by using the simulated annealing method, and the combinations of the high-temperature-resistant samples of the first category with the number of SNP sites related to the high-temperature-resistant trait and the homozygosity greater than a preset value are obtained.

[0028] In the process of calculating the genetic force, the combination of the high-temperature-resistant samples of the first type is avoided from inbreeding, that is, the high-temperature-resistant samples of the first type in the combination of the high-temperature-resistant samples of the first type are prevented from being genetically related within three generations;

[0029] According to the obtained genetic force of the high-temperature-resistant samples of the first type, the high-temperature-resistant samples of the first type with a genetic force greater than a preset value are determined, that is, the number of SNP sites related to the high-temperature-resistant traits and the homozygosity of the corresponding high-temperature-resistant samples of the first type when combined with other high-temperature-resistant samples of the first type are greater than the preset value, and the high-temperature-resistant samples of the first type are labeled as core breeding samples, and the number of the core breeding samples is controlled to be greater than a predetermined number.

[0030] Further, in a preferred embodiment of the present application, the core breeding samples are combined and bred, the heat-resistant related genes in the core breeding samples are aggregated, and generation selection processing is performed, so that the high-temperature-resistant traits of the new variety of Pseudosciaena crocea of the Naozhou population are fixed, and specifically:

[0031] The tag information of the core breeding samples is updated, and the number of SNP sites related to the corresponding high-temperature-resistant traits and the homozygosity are recorded in the tag information of the core breeding samples;

[0032] A breeding and culture tank is set up, and the core breeding samples are put into the breeding and culture tank and bred based on the high-heritability x high-heritability principle, wherein the high-heritability x high-heritability principle is to sort all the core breeding samples in descending order of genetic force, and to preferentially select core breeding samples with high genetic force and non-close relatives for breeding and pairing processing;

[0033] After the core breeding samples are bred and paired, breeding fry of different combinations are obtained, all the breeding fry of different combinations are saved in a breeding and culture tank for breeding, and the core breeding samples are captured according to the positioning of the core breeding samples recorded on the electronic tags of the core breeding samples, so that only hybrid breeding fry are left in the breeding and culture tank;

[0034] During the breeding process, the water quality conditions and environmental conditions that enable the fry to avoid death are searched based on a big data network and outputted;

[0035] After the breeding is completed, the number of SNP sites related to the high-temperature-resistant traits and the homozygosity in the fry are determined by performing gene extraction analysis on the breeding fry of different combinations;

[0036] During the breeding process, the tag information of the core breeding samples is labeled as the parent information of the fry, and the complete pedigree of the breeding population of different combinations is established by combining the parent information of the fry and the number of SNP sites related to the heat-resistant traits and the homozygosity of the fry, and the complete pedigree of the population is used for generation selection processing, so that the target high-temperature-resistant new variety of Pseudosciaena crocea of the Naozhou population is obtained.

[0037] Further, in a preferred embodiment of the present application, the population complete pedigree of different combinations is established, and the population complete pedigree is used for generation selection processing to obtain a target high-temperature-resistant new variety of Pseudosciaena crocea of Naozhou population, specifically:

[0038] Within the breeding population complete pedigree, the juvenile fish of different combinations is labeled as the F1 generation population;

[0039] The F1 generation population continues to be subjected to high-temperature stress experiments, and after the high-temperature stress experiments, the F1 generation population with a genetic force greater than a preset value is selected as the parent of the next generation according to the genetic force of the F1 generation population, and the pedigree of the breeding population of different combinations is updated;

[0040] Based on the breeding population complete pedigree of different combinations, the crossbreeding of Pseudosciaena crocea of Naozhou population is repeatedly performed to obtain F2, F3…Fn generations, and in the crossbreeding process, the phenotype information of each generation is measured, and the number of heat-resistant related SNP sites and the calculation of homozygosity are performed, and in the breeding process, the population with high genetic force and non-close relatives is preferentially selected for breeding pairing processing;

[0041] A preset selection target generation number is set, if the selected generation number is equal to the selection target generation number, the generation selection processing is stopped, and different combination breeding juvenile fish to be analyzed is obtained;

[0042] The core indicators of the different combination breeding juvenile fish to be analyzed are measured, if the core indicators of the breeding juvenile fish to be analyzed meet the preset indicator threshold, the breeding juvenile fish is labeled as a target heat-resistant new variety of Pseudosciaena crocea of Naozhou population, if not, the combination breeding processing is continued until the core indicators of the breeding juvenile fish to be analyzed meet the preset indicator threshold.

[0043] The system for breeding a new high-temperature-resistant variety of Pseudosciaena crocea of Naozhou population is characterized in that the breeding system integrates a high-performance computing architecture and a bioinformatics storage module, and includes a non-volatile memory composed of an ECC-verified DDR4 RDIMM memory module and an NVMe solid-state storage array using 3D NAND flash, and a multi-core processor based on Zen4 microarchitecture; the memory is solidified and deployed with a breeding method program having a phenotype-genotype correlation analysis engine, and when the program is executed in parallel by the superscalar pipeline execution unit in the processor, the following steps are implemented:

[0044] Sample collection is performed on Pseudosciaena crocea of Naozhou population, and after sample collection, high-temperature stress experiments are performed to achieve the purpose of screening high-temperature-resistant samples;

[0045] Parental genetic evaluation and analysis are performed on the high-temperature-resistant samples, the genetic potential of the high-temperature-resistant samples is calculated, and core breeding samples are screened from the high-temperature-resistant samples;

[0046] The core breeding sample is hybridized, the heat-resistant related genes in the core breeding sample are aggregated, and generation selection is processed, so that the high-temperature-resistant trait of the new Pseudosciaena crocea of Naozhou population is fixed.

[0047] The technical defects in the background art are solved by the present application, and the present application has the following beneficial effects: high-temperature stress experiments are performed on the cultured Pseudosciaena crocea of Naozhou population, and heat-resistant samples are screened based on survival rate and heat stress physiological indexes; the genetic potential of the heat-resistant samples is evaluated by using parent genetic evaluation, so as to achieve the purpose of screening core breeding samples. Finally, the breeding is carried out by designing a low inbreeding pairing scheme, the heat-resistant advantage related genes are aggregated, and the high-temperature-resistant trait is fixed through continuous selection. The present application can break through the bottleneck of the death of Pseudosciaena crocea of Naozhou population under high temperature, and obtain a new high-temperature-resistant variety of Pseudosciaena crocea of Naozhou population, which provides sustainable seed source support for expanding the culture space of Pseudosciaena crocea and the seawater fish culture industry under the background of warming. BRIEF DESCRIPTION OF DRAWINGS

[0048] In order to more clearly illustrate the technical solutions in the embodiments of the present application or the prior art, the following will briefly introduce the drawings needed to be used in the embodiments or the prior art description. Obviously, the drawings in the following description are only some embodiments of the present application, and other drawings of other embodiments can also be obtained by those skilled in the art without any creative effort.

[0049] Figure 1 A flowchart of the breeding method of the high-temperature-resistant new variety of Pseudosciaena crocea of Naozhou population is shown;

[0050] Figure 2 A flowchart of the method for calculating the genetic potential of the heat-resistant sample and screening the core breeding sample in the heat-resistant sample is shown;

[0051] Figure 3 A program view of the breeding system of the high-temperature-resistant new variety of Pseudosciaena crocea of Naozhou population is shown. DETAILED DESCRIPTION

[0052] In order to more clearly understand the above-mentioned purposes, features and advantages of the present application, the present application will be further described in detail below in combination with the drawings and specific embodiments. It should be noted that the embodiments of the present application and the features in the embodiments can be combined with each other without conflict.

[0053] In the following description, many specific details are set forth in order to provide a thorough understanding of the present application, however, the present application can also be implemented in other ways different from those described herein, therefore, the protection scope of the present application is not limited by the specific embodiments disclosed below.

[0054] Figure 1A flow chart of a method for breeding a new high-temperature-resistant variety of Pseudosciaena crocea of the Xizhou population is shown, including the following steps:

[0055] S102: Collect samples of Pseudosciaena crocea of the Xizhou population from different sources, and perform a high-temperature stress experiment after sample collection to achieve the purpose of screening high-temperature-resistant samples;

[0056] S104: Perform parent genetic evaluation analysis on the high-temperature-resistant samples, calculate the genetic potential of the high-temperature-resistant samples, and screen core breeding samples from the high-temperature-resistant samples;

[0057] S106: Perform combination design breeding on the core breeding samples, aggregate heat-resistant related genes within the core breeding samples, and perform generation breeding processing to fix the high-temperature-resistant traits of the Pseudosciaena crocea of the Xizhou population.

[0058] Further, in a preferred embodiment of the present application, the purpose of collecting samples of Pseudosciaena crocea of the Xizhou population from different sources and performing a high-temperature stress experiment after sample collection to achieve the purpose of screening high-temperature-resistant samples is specifically:

[0059] Collect Pseudosciaena crocea of the Xizhou population from different farms, and control the collected Pseudosciaena crocea of the Xizhou population to cover different families, wherein the family is Pseudosciaena crocea of the Xizhou population with mating behavior and parentage index, and the Pseudosciaena crocea of the Xizhou population is labeled as a target sample;

[0060] Record the phenotype information of all collected target samples, including the age, size, and fish health status of the target samples;

[0061] Randomly group all target samples, and set different high-temperature breeding temperatures for target samples in different groups to perform gradient high-temperature stress experiments, and set a control group, wherein the breeding temperature of the target samples in the control group is the standard breeding temperature of the target samples;

[0062] During the gradient high-temperature stress experiment, real-time record the core indicators of target samples in different groups, wherein the core indicators include the survival rate, heat-resistant time, and physiological and biochemical indicators of the target samples;

[0063] The heat-resistant time is the time from the beginning of stress to death of the target sample, or the time from the beginning of stress to the appearance of heat stress symptoms;

[0064] Among different groups of target samples, combined with the corresponding core indicators, screen the target samples with the highest survival rate and the longest heat-resistant time in different groups, and label them as high-temperature-resistant samples.

[0065] It should be noted that, in order to breed new high-temperature-resistant varieties of Pseudosciaena crocea, sample extraction needs to be performed first. Because the number of Pseudosciaena crocea in the breeding pond is large, the high-temperature-resistant gene needs to be aggregated by extracting samples. The Pseudosciaena crocea samples collected should be from different breeding farms or families, because close crossbreeding can easily cause variation of heat-resistant genes, making it impossible to breed new high-temperature-resistant varieties of Pseudosciaena crocea. The phenotypic information of all target samples collected is recorded to facilitate observation of changes in Pseudosciaena crocea during the breeding of new varieties. In the gradient high-temperature stress experiment, Pseudosciaena crocea is randomly divided into several groups and exposed to different high temperatures, such as 30°C and 33°C. The temperature gradient needs to be accurately controlled, and the stress time needs to be long enough until the Pseudosciaena crocea shows a clear death inflection point, and the core indicators of the target samples in different groups are recorded in real time. Physiological and biochemical indicators include but are not limited to indicators related to heat resistance, such as heat shock protein expression, antioxidant enzyme activity, serum cortisol level, and changes in gill / liver tissue structure. Heat stress symptoms include imbalance, lateral turning, and rapid breathing of Pseudosciaena crocea.

[0066] Further, in a preferred embodiment of the present application, the core breeding samples are crossed and bred, the heat-resistant related genes in the core breeding samples are aggregated, and generation selection is performed to fix the high-temperature-resistant traits of Pseudosciaena crocea, specifically:

[0067] The tag information of the core breeding samples is updated to record the number of heat-resistant related SNP sites and the homozygosity in the tag information of the core breeding samples.

[0068] A breeding pond is set up, and the core breeding samples are put into the breeding pond and bred based on the high heritability x high heritability principle, wherein the high heritability x high heritability principle is to sort all core breeding samples in descending order of heritability, and to preferentially select core breeding samples with high heritability and non-close relatives for breeding pairing.

[0069] After the core breeding samples are bred and paired, juvenile fish of different combinations are obtained, all combined juvenile fish are saved in the breeding pond for breeding, and the core breeding samples are captured based on the positioning of the core breeding samples recorded on the electronic tags implanted in the core breeding samples, so that only hybrid breeding juvenile fish are left in the breeding pond.

[0070] During the breeding process, the water quality conditions and environmental conditions that can avoid the death of juvenile fish are searched based on a big data network and output.

[0071] After the breeding is completed, the heat-resistant related SNP sites and the homozygosity in the juvenile fish are determined by extracting and analyzing the genes of the juvenile fish of different combinations.

[0072] In the breeding process, the label information of the core breeding sample is calibrated as the parent information of the juvenile fish. The complete pedigree of the breeding population is established by combining the parent information of the juvenile fish, the number of heat tolerance related SNP sites and the homozygosity of the juvenile fish. The generation selection breeding process is carried out according to the complete pedigree of the breeding population of different combinations, and the target high-temperature-resistant new variety of Pseudosciaena crocea of Naozhou population is obtained.

[0073] It should be noted that the breeding and breeding pool is set up for the combined pairing breeding process of the screened core breeding sample, and based on the high heritability x high heritability principle, the obtained juvenile fish also has high heritability, which is convenient for obtaining the new high-temperature-resistant variety. According to the label positioning, the core breeding sample is captured in the breeding and breeding pool to reduce the possibility of inbreeding and ensure that the water quality and environmental conditions of the breeding pool are suitable for juvenile fish breeding. After breeding, the juvenile fish is continuously subjected to genetic analysis, and the complete pedigree of the breeding population is constructed for the breeding of the new high-temperature-resistant variety.

[0074] Further, in a preferred embodiment of the present application, the complete pedigree of the breeding population is established, and the generation selection breeding process is carried out according to the complete pedigree of the breeding population of different combinations, and the target high-temperature-resistant new variety of Pseudosciaena crocea of Naozhou population is obtained, specifically:

[0075] In the complete pedigree of the breeding population, the breeding juvenile fish of the first combination is calibrated as the F1 generation population;

[0076] The F1 generation population is continuously subjected to high temperature stress experiment, and according to the heritability of the F1 generation population after the high temperature stress experiment, the F1 generation population with heritability greater than the preset value is selected as the parent of the next generation, and the pedigree of the complete pedigree of the breeding population is updated;

[0077] Based on the complete pedigree of the breeding population of different combinations, the combination design breeding process of Pseudosciaena crocea of Naozhou population is repeatedly carried out to obtain F2, F3…Fn generation population, and in the breeding process, the phenotype information of each generation is measured, and the number of heat tolerance related SNP sites and homozygosity are calculated, and in the breeding process, the population with high heritability and non-close relatives is preferentially selected for breeding pairing process;

[0078] The number of breeding generations is preset, and if the number of breeding generations is equal to the number of breeding target generations, the generation selection breeding process is stopped, and the breeding combination juvenile fish to be analyzed is obtained;

[0079] The core index of the breeding combination juvenile fish to be analyzed is measured, and if the core index of the breeding combination juvenile fish to be analyzed meets the preset index threshold, the breeding combination juvenile fish to be analyzed is calibrated as the target new high-temperature-resistant variety of Pseudosciaena crocea of Naozhou population, and if it does not meet the requirement, the combination design breeding process is continuously carried out until the core index of the obtained breeding combination juvenile fish to be analyzed meets the preset index threshold.

[0080] It should be noted that the overall heat tolerance is evaluated by the method of generation breeding, repeated high temperature stress experiment, and excellent heat-resistant sample individuals are screened according to the phenotype data for further breeding of the next generation. After the breeding generation number is equal to the target generation number, the genetic performance is stable, the high temperature resistance is significantly improved, and a new strain with excellent comprehensive traits is obtained. Then, the core indicators are judged, and if the standard conditions are met, the new heat-resistant Pseudosciaena crocea variety of the Zhizhou population can be obtained.

[0081] Figure 2 A flow chart of a method for calculating the genetic potential of heat-resistant samples and screening core breeding samples in heat-resistant samples is shown, including the following steps:

[0082] S202: Parental genetic evaluation analysis is performed on the heat-resistant samples, the genetic potential of the heat-resistant samples is calculated, and the core breeding samples are screened from the heat-resistant samples;

[0083] S204: The genetic parameters of all heat-resistant samples of the first type are evaluated in combination with the label information of the heat-resistant samples of the first type and the corresponding heat-resistant related SNP sites, and the core breeding samples are screened according to the evaluation results.

[0084] Further, in a preferred embodiment of the present application, the parental genetic evaluation analysis of the heat-resistant samples, the calculation of the genetic potential of the heat-resistant samples, and the screening of the core breeding samples from the heat-resistant samples are specifically:

[0085] Subcutaneous electronic tags are implanted in all heat-resistant samples, and the label information recorded in the implanted electronic tags is the phenotype information of the corresponding heat-resistant samples and the implantation date;

[0086] The implanted electronic tags can be used to locate the heat-resistant samples during mating of different heat-resistant samples;

[0087] Tail fin tissues are cut from all heat-resistant samples, and DNA extraction treatment is performed on the cut tail fin tissues of the heat-resistant samples by using a DNA kit to obtain heat-resistant sample DNA;

[0088] The heat-resistant sample DNA is subjected to PCR amplification, and a big data network is introduced to analyze the target gene effect of the PCR-amplified heat-resistant samples, determine the SNP sites related to the heat-resistant traits, and mark them as heat-resistant trait-related SNP sites;

[0089] genotyping all high-temperature-resistant samples, calculating the number of high-temperature-resistant trait-related SNP sites in the high-temperature-resistant samples, ranking all high-temperature-resistant samples in descending order according to the number of high-temperature-resistant trait-related SNP sites, and calculating the homozygosity of high-temperature-resistant trait-related SNP sites in different high-temperature-resistant samples, and then ranking the high-temperature-resistant samples again according to the homozygosity, and outputting a preliminary genetic potential ranking table of the high-temperature-resistant samples;

[0090] According to the preliminary genetic potential ranking table of the high-temperature-resistant samples, high-temperature-resistant samples ranked in the preset top are screened and labeled as high-temperature-resistant samples of the first type.

[0091] According to the preliminary genetic potential ranking table of the high-temperature-resistant samples, high-temperature-resistant samples ranked in the preset top are screened and labeled as high-temperature-resistant samples of the first type.

[0092] It should be noted that all high-temperature-resistant samples are implanted with subcutaneous electronic tags to facilitate the positioning of the location of the high-temperature-resistant samples, because the high-temperature-resistant samples need to be salvaged in the breeding tank later to prevent inbreeding. Then the DNA of the high-temperature-resistant samples needs to be extracted, and the DNA extraction is performed using a DNA kit, and the tail fin tissue is cut off first, and ethanol is used for fixation to realize the sample collection of DNA. The purpose of PCR amplification of the DNA of the high-temperature-resistant samples is to amplify the number of genes, increase the number of gene samples, and facilitate the extraction of SNP sites. After amplification, the target genes related to heat resistance are extracted and labeled as high-temperature-resistant trait-related SNP sites. The heat-resistant related genes need to be typed, because different heat-resistant related genes have different heat-resistant effects, so their genetic potential is also different. Some genes can be inherited and have good heat-resistant effect, and are suitable for new variety breeding. According to the number of high-temperature-resistant trait-related SNP sites and the size of homozygosity, the heat-resistant traits of the samples are judged, and high-temperature-resistant samples of the first type are screened.

[0093] Further, in a preferred embodiment of the present application, the genetic parameter evaluation of all high-temperature-resistant samples of the first type is performed in combination with the tag information of the high-temperature-resistant samples of the first type and the corresponding high-temperature-resistant trait-related SNP sites, and the core breeding samples are screened according to the evaluation results.

[0094] The tag information of the high-temperature-resistant samples of the first type and the corresponding high-temperature-resistant trait-related SNP sites are imported into the animal model analysis software, and the genetic force calculation of all high-temperature-resistant samples of the first type is performed by the animal model analysis software.

[0095] The genetic force calculation is that a high-temperature-resistant sample is combined freely and a kinship matrix is constructed, and the number of SNP sites related to the high-temperature-resistant trait and the homozygosity of the juvenile fish obtained by further combining and mating different combinations of the high-temperature-resistant sample are predicted by means of the simulated annealing method, and a high-temperature-resistant sample combination in which the number of SNP sites related to the high-temperature-resistant trait and the homozygosity are greater than preset values is obtained,

[0096] In the process of genetic force calculation, inbreeding of the high-temperature-resistant sample combination is avoided, that is, the high-temperature-resistant samples existing in the high-temperature-resistant sample combination are not in three generations of inheritance.

[0097] According to the obtained genetic force of the high-temperature-resistant sample, a high-temperature-resistant sample with a genetic force greater than a preset value is determined, that is, the number of SNP sites related to the high-temperature-resistant trait and the homozygosity obtained by combining the corresponding high-temperature-resistant sample with other high-temperature-resistant samples are greater than the preset values, and the high-temperature-resistant sample is labeled as a core breeding sample, and the number of core breeding samples is controlled to be greater than a predetermined number.

[0098] It should be noted that the animal model analysis software is a software for predicting the genetic force of a sample, and the genetic force of the sample can be estimated by importing the label information of the high-temperature-resistant sample and the corresponding SNP sites related to the high-temperature-resistant trait into the software, that is, the number of SNP sites related to the high-temperature-resistant trait and the homozygosity of the juvenile fish obtained by further combining and mating different combinations of the high-temperature-resistant sample. The simulated annealing algorithm combined with the software for genetic force estimation can output the optimal solution of the prediction. Inbreeding needs to be avoided to ensure sufficient genetic diversity, so a large number of effective samples are needed.

[0099] As shown in Figure 3 The second aspect of the present application also provides a breeding system for a new high-temperature-resistant variety of Pseudosciaena crocea in Naozhou, characterized in that the breeding system integrates a high-performance computing architecture and a bioinformatics storage module, including a non-volatile memory composed of an ECC-verified DDR4 RDIMM memory module and an NVMe solid-state storage array using 3D NAND flash, and a multi-core processor based on Zen4 microarchitecture; the memory is solidified and deployed with a breeding method program having a phenotype-genotype correlation analysis engine, and when the program is executed in parallel by the superscalar pipeline execution unit in the processor, the following steps are realized:

[0100] Samples of Pseudosciaena crocea in Naozhou are collected, and a high-temperature stress experiment is performed after sample collection to achieve the purpose of screening high-temperature-resistant samples;

[0101] The parent genetic evaluation and analysis of the high-temperature-resistant samples are performed, the genetic potential of the high-temperature-resistant samples is calculated, and the core breeding samples are screened from the high-temperature-resistant samples;

[0102] The core breeding sample is hybridized, the heat-resistant related genes in the core breeding sample are aggregated, and generation selection is processed, so that the high-temperature-resistant trait of the Oryzias genus Pseudosciaena polyactis is fixed.

[0103] The above merely illustrates the specific embodiments of the present application, but the protection scope of the present application is not limited thereto, any person skilled in the art can easily think of the changes or replacements within the technical range disclosed by the present application, which should be covered in the protection scope of the present application. Therefore, the protection scope of the present application should be subject to the protection scope of the claims.

Claims

1. A method for breeding new high-temperature tolerant varieties of large yellow croaker from Naozhou, characterized by: Includes the following steps: Samples were collected from large yellow croaker of the Naozhou group, and high-temperature stress experiments were conducted after sample collection to screen for high-temperature resistant samples. Genetic evaluation analysis of the parents of heat-resistant samples was performed to calculate the genetic potential of the heat-resistant samples and to select core breeding samples from the heat-resistant samples. The core breeding samples were paired and combined to aggregate heat-resistant genes within the core breeding samples, and then subjected to generational selection to fix the heat-resistant trait of the resulting new variety of Naozhou large yellow croaker.

2. The method for breeding a new high-temperature resistant variety of large yellow croaker from the Naozhou group according to claim 1, characterized in that, The process of collecting samples from large yellow croaker of the Naozhou group and conducting high-temperature stress experiments after sample collection aims to screen for high-temperature resistant samples. Specifically: Samples of large yellow croaker from Naozhou were collected from a farm, and the collected samples were controlled to cover different families. Among them, the families were large yellow croaker samples with mating behavior and parentage indicators, and the samples were labeled as target samples. Record the phenotypic information of all collected target samples, including the age, size, and health status of the fish. All target samples were randomly grouped, and different high-temperature culture temperatures were set for different groups of target samples to conduct gradient high-temperature stress experiments. At the same time, a control group was set up, in which the culture temperature of the target samples in the control group was the standard culture temperature of the target samples. During the gradient high temperature stress experiment, the core indicators of the target samples in different groups were recorded in real time. The core indicators included the survival rate, heat resistance time and physiological and biochemical indicators of the target samples. The heat resistance time is the time from the onset of stress to death of the target sample, or it can be the time from the onset of stress to the appearance of heat stress symptoms. Within different groups of target samples, based on the corresponding core indicators, the target samples with the highest survival rate and the longest heat resistance time in each group were selected and labeled as high-temperature resistant samples.

3. The method for breeding a new high-temperature resistant variety of large yellow croaker from the Naozhou group according to claim 1, characterized in that, The process of conducting parental genetic evaluation analysis on heat-resistant samples, calculating the genetic potential of the heat-resistant samples, and selecting core breeding samples from the heat-resistant samples specifically involves: All heat-resistant samples underwent subcutaneous electronic tag implantation, and the tag information recorded in the implanted electronic tag was the phenotypic information of the corresponding heat-resistant sample and the implantation date; Among them, the implanted electronic tag can locate the heat-resistant sample during the mating of different heat-resistant samples; Caudal fin tissue was excised from all heat-resistant samples, and DNA was extracted from the excised caudal fin tissue using a DNA kit to obtain heat-resistant sample DNA. DNA from heat-resistant samples was amplified by PCR and introduced into a big data network for target gene action analysis of the PCR-amplified heat-resistant samples. This identified SNP sites associated with heat-resistant traits and labeled them as heat-resistant trait-related SNP sites. Genotyping was performed on all heat-resistant samples. The genotyping process involved calculating the number of SNP loci related to heat resistance in the heat-resistant samples, sorting all heat-resistant samples in reverse order based on the number of SNP loci related to heat resistance, calculating the homozygosity of SNP loci related to heat resistance in different heat-resistant samples, and sorting the heat-resistant samples a second time based on the homozygosity. The preliminary genetic potential ranking table of heat-resistant samples was then output. Based on the preliminary genetic potential ranking table of the heat-resistant samples, the heat-resistant samples ranked at the top of the preset ranking are screened and labeled as a type of heat-resistant sample. By combining the tag information of a class of heat-resistant samples with the corresponding SNP loci related to heat-resistant traits, the genetic parameters of all class of heat-resistant samples were evaluated, and core breeding samples were selected based on the evaluation results.

4. The method for breeding a new high-temperature resistant variety of large yellow croaker from the Naozhou group according to claim 3, characterized in that, The genetic parameters of all heat-resistant samples were evaluated by combining the tag information of a class of heat-resistant samples with the corresponding SNP loci related to heat-resistant traits. Core breeding samples were then selected based on the evaluation results. Specifically: Animal model analysis software was introduced, and the tag information of a class of heat-resistant samples and the corresponding SNP loci related to the heat-resistant trait were imported into the animal model analysis software. Heritability calculation was performed on all the class of heat-resistant samples through the animal model analysis software. The heritability calculation involves freely assorting a class of heat-resistant samples and constructing a kinship matrix. Then, using simulated annealing, it predicts the number and homozygosity of heat-resistant SNP loci in juvenile fish obtained from further mating of different combinations of these heat-resistant samples. The goal is to identify a class of heat-resistant sample combinations where both the number of heat-resistant SNP loci and the homozygosity are greater than a preset value. During the calculation of heritability, inbreeding of a heat-resistant sample combination should be avoided, that is, the heat-resistant samples within a heat-resistant sample combination should be avoided to be inherited within three generations. Based on the heritability of the obtained heat-resistant sample, a heat-resistant sample with a heritability greater than the preset value is identified. That is, when the corresponding heat-resistant sample is combined with other heat-resistant samples, the number of heat-resistant trait-related SNP sites and the homozygosity are both greater than the preset value. This sample is then designated as a core breeding sample, and the number of core breeding samples is controlled to be greater than the predetermined number.

5. The method for breeding a new high-temperature resistant variety of large yellow croaker from the Naozhou group according to claim 1, characterized in that, The process involves pairing and combining core breeding samples, aggregating heat-resistant genes within these samples, and then performing generational selection to fix the heat-resistant trait in the resulting new variety of Naozhou large yellow croaker. Specifically: The tag information of the core breeding samples is updated so that the tag information of the core breeding samples records the number of SNP sites related to the heat resistance trait and the homozygosity. A breeding and breeding pond is set up. In the breeding and breeding pond, based on the principle of high heritability × high heritability, core breeding samples are put in and bred and paired. The principle of high heritability × high heritability is to sort all core breeding samples in reverse order of heritability and to select core breeding samples with high heritability and that are not closely related for breeding and pairing. After the core breeding sample is bred and paired, different breeding combinations of juvenile fish are obtained. All the breeding combination juvenile fish are stored in the breeding and breeding pond for breeding treatment. The core breeding sample is harvested according to the location of the core breeding sample recorded on the electronic tag implanted in the core breeding sample, so that only different breeding combination juvenile fish are in the breeding and breeding pond. During the breeding process, based on big data networks, the water quality and environmental conditions that prevent juvenile fish from dying in different breeding combinations are retrieved and output; After the breeding was completed, gene extraction and analysis were performed on the juvenile fish of different breeding combinations to determine the number and homozygosity of SNP sites related to heat resistance in the juvenile fish of different breeding combinations. During the breeding process, the tag information of the core breeding sample was marked as the parent information of the juvenile fish of different breeding combinations. Combined with the parent information of the juvenile fish of different combinations and the number and homozygosity of SNP loci related to the heat resistance trait, a complete pedigree of different breeding groups was established. Based on the complete pedigree of the breeding groups, generational selection was carried out to obtain a new heat-resistant variety of Naozhou large yellow croaker.

6. The method for breeding a new high-temperature resistant variety of large yellow croaker from the Naozhou group according to claim 5, characterized in that, The process of establishing a complete pedigree for the breeding population and conducting generational selection based on this pedigree to obtain a new heat-resistant variety of large yellow croaker from the Naozhou group is as follows: Within the complete pedigree of the breeding population, juvenile fish of a specific breeding combination are identified as the F1 generation. High temperature stress experiments were continued on the F1 generation population. After the high temperature stress experiments, based on the heritability of the F1 generation population, the F1 generation population with heritability greater than the preset value was selected as the parents of the next generation. At the same time, the complete pedigree of the breeding population was updated. Based on the complete pedigree of the breeding population, different breeding combinations of Naozhou large yellow croaker were repeatedly performed to obtain F2, F3...Fn generation populations. During the breeding process, phenotypic information was measured for each generation, and the number and homozygosity of SNP loci related to heat resistance traits were calculated. At the same time, during the breeding process, it is necessary to prioritize the selection of populations with high heritability and that are not closely related for pairing. The target generation for breeding is preset. If the number of generations selected is equal to the target generation, the generation selection process is stopped, and the juvenile fish to be analyzed are obtained. The core indicators of the juvenile fish to be analyzed are determined. If the core indicators of the juvenile fish to be analyzed meet the preset indicator thresholds, the juvenile fish to be analyzed are identified as the target high-temperature resistant new variety of large yellow croaker from Naozhou. If they do not meet the thresholds, the breeding process continues until the core indicators of the juvenile fish to be analyzed meet the preset indicator thresholds.

7. A breeding system for a new high-temperature resistant variety of large yellow croaker from Naozhou, characterized by: The breeding system integrates a high-performance computing architecture and a bioinformatics storage module, including a non-volatile memory consisting of a DDR4 RDIMM memory module with ECC verification and an NVMe solid-state storage array using 3D NAND flash memory, as well as a multi-core processor based on the Zen4 microarchitecture. The memory contains a breeding method program with a phenotypic-genotypic association analysis engine. When the program is decoded and executed in parallel by the superscalar pipeline execution unit in the processor, it implements the breeding method steps as described in any one of claims 1-6, and accelerates the calculation of large-scale SNP sites through the matrix operation unit integrated in the processor.

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