CN116814798B - Molecular marker for breeding rapid growth type largemouth black bass and application thereof - Google Patents
Molecular marker for breeding rapid growth type largemouth black bass and application thereof Download PDFInfo
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Abstract
The invention provides a molecular marker for breeding of quick-growth micropterus salmoides and application thereof, belonging to the technical field of aquatic breeding. The invention screens the fast-growth related markers in the American stock F1 generation by simplifying genome sequencing, verifies the relevance of the enrichment and growth performance of the fast-growth markers, and utilizes the developed markers to combine with corresponding verification and matching technologies to complete the continuous breeding work of new varieties. Experiments prove that SNP19140160, SNP23355498, SNP9639603 and SNP9639605 are obviously related to the determination of the F1 generation body length of the stock of the micropterus salmoides in the experiments; and the SNP19140160 locus is obviously related to the weight, height and thickness of the F1 generation of the largemouth black bass stock. The molecular marker provided by the invention can accurately and efficiently screen and identify the rapid growth type micropterus salmoides, so that the breeding efficiency of novel varieties of the rapid growth type micropterus salmoides is improved.
Description
Technical Field
The invention belongs to the technical field of aquatic breeding, and particularly relates to a molecular marker for breeding of rapid growth type micropterus salmoides and application thereof.
Background
In the culture process of the largemouth bass, the survival rate of the feedback seedlings of a plurality of farmers is low, and diseases are easy to occur. Combining genetic characteristics and stress characteristics of micropterus salmoides, low genetic diversity of parents and poor stress resistance of offspring may be key problems in limiting the development of industry. Due to serious germplasm degradation, the disease resistance of the cultured micropterus salmoides is drastically reduced, so that diseases are frequently generated in recent years. Therefore, the breeding of the novel largemouth bass variety is not only helpful for solving the problem of parent germplasm degeneration, but also can relieve the problem of reduced culture benefits of fries and adult fishes caused by lack of improved varieties. The Chinese aquatic science institute freshwater fishery research center introduces 7000 tail stock seedlings of northern subspecies of micropterus salmoides from the U.S. The stock seeds have clear genetic pedigree and high genetic diversity, and the parents used are subjected to biosafety detection by organic certification of the United states department of agriculture and do not carry any viruses. Based on the genetic information of 11 microsatellite polymorphic loci, the genetic distance between the northern subspecies of the micropterus salmoides and the dominant cultured Lateolabrax japonicus No. 1 and the Taiwan strain Lateolabrax japonicus in the current market is found to be far, the genetic diversity is high, and the method has good breeding potential.
The system is used for developing the stock preservation and breeding work of the micropterus salmoides, is an important way for solving the quality degradation of the parent of the micropterus salmoides at present, and is also a necessary measure for improving the yield of the micropterus salmoides and increasing the income of farmers. Molecular marker assisted breeding (MARKER ASSISTED Selection, MAS) can effectively overcome the defects of the traditional breeding method, and can accurately and efficiently achieve the purpose of breeding. The method screens target characters of selected individuals by utilizing molecular markers closely linked or representing co-segregation with target genes, so that linkage encumbrance is reduced, expected individuals are obtained, and the purpose of improving breeding efficiency is achieved.
However, there is no molecular marker for breeding micropterus salmoides with rapid growth characteristics, which undoubtedly hinders the breeding work of micropterus salmoides.
Disclosure of Invention
In view of the above, the invention aims to provide a molecular marker for breeding the rapid-growth micropterus salmoides, which is closely linked with the rapid-growth index, and is a good molecular marker for breeding a novel variety of the rapid-growth micropterus salmoides, so that the problem of germplasm degeneration of the micropterus salmoides is solved.
The invention provides a molecular marker for breeding quick-growth micropterus salmoides, which comprises the following SNP molecular markers; SNP19140160, SNP23355498, SNP9639603, and SNP9639605;
The SNP19140160 is a site A/C with polymorphism at the position 19140160 of NW_024044570.1 chromosome in REFSEQ GCF _014851395.1 gene version;
the SNP23355498 is a polymorphic site G/A at the position 23355498 of NW_024044348.1 in REFSEQ GCF _014851395.1 gene version;
The SNP9639603 is the existence of a polymorphic site T/G at the position 9639603 of the NW_024044237.1 chromosome in REFSEQ GCF _014851395.1 gene version;
The SNP9639605 is the presence of a polymorphic site T/A at position 9639605 of chromosome NW_024044237.1 in the REFSEQ GCF _014851395.1 gene version.
Preferably, the SNP19140160 is located at position 100 in the sequence shown in SEQ ID NO. 3
Polymorphic site A/C at the position.
Preferably, the SNP23355498 is a polymorphic site G/A at position 100 in the sequence shown in SEQ ID NO. 13.
Preferably, the SNP9639603 is a polymorphic site T/G at position 100 in the sequence shown in SEQ ID NO. 10.
Preferably, the SNP9639605 is a polymorphic site T/A at position 100 in the sequence shown in SEQ ID NO. 12.
The invention provides a primer for detecting quick-growing micropterus salmoides, which comprises a primer pair for amplifying the molecular markers;
the primer pair for amplifying SNP19140160 molecular markers comprises a forward primer with a nucleotide sequence shown as SEQ ID NO. 18 and a reverse primer with a nucleotide sequence shown as SEQ ID NO. 19;
the primer pair for amplifying SNP23355498 molecular markers comprises a forward primer with a nucleotide sequence shown as SEQ ID NO. 36 and a reverse primer with a nucleotide sequence shown as SEQ ID NO. 37;
The primer pair for amplifying SNP9639603 molecular markers comprises a forward primer with a nucleotide sequence shown as SEQ ID NO. 32 and a reverse primer with a nucleotide sequence shown as SEQ ID NO. 33;
The primer pair for amplifying SNP9639605 molecular markers comprises a forward primer with a nucleotide sequence shown as SEQ ID NO. 34 and a reverse primer with a nucleotide sequence shown as SEQ ID NO. 35.
The invention provides a kit for detecting rapid growth type micropterus salmoides, which comprises the primer and a PCR reaction solution.
The invention provides application of the molecular marker, the primer pair or the kit in auxiliary breeding of a fast-growing micropterus salmoides variety.
Preferably, the auxiliary breeding method of the rapid growth type largemouth bass variety comprises the following steps:
And amplifying a molecular marker of the sample to be bred by using the primer, wherein the sample to be bred is used as a material for breeding when the genome DNA of the sample to be bred is enriched in the dominant growth genotype of the molecular marker and the growth speed of the sample to be bred is improved by more than 12% compared with that of a non-bred population.
Preferably, the dominant growth genotypes of the molecular markers include more than 2 of the following SNP molecular marker dominant growth genotypes:
the dominant growth genotype of SNP19140160 is AA;
the dominant growth genotype of SNP9639603 is GG or TT;
the dominant growth genotype of SNP9639605 is AA;
the dominant growth genotype of SNP23355498 is AA or GG.
The invention provides a molecular marker for breeding quick-growth micropterus salmoides, which comprises at least one SNP molecular marker as follows; SNP19140160, SNP23355498, SNP9639603, and SNP9639605; the SNP19140160 is a site A/C with polymorphism at the position 19140160 of NW_024044570.1 chromosome in REFSEQ GCF _014851395.1 gene version; the SNP23355498 is a polymorphic site G/A at the position 23355498 of NW_024044348.1 in REFSEQ GCF _014851395.1 gene version; the SNP9639603 is the existence of a polymorphic site T/G at the position 9639603 of the NW_024044237.1 chromosome in REFSEQ GCF _014851395.1 gene version; the SNP9639605 is REFSEQ GCF-014851395.1 gene version
A polymorphic site T/A exists at the 9639605 position of NW_ 024044237.1. The SNPs 19140160, 23355498, 9639603 and 9639605 are closely linked to the growth trait. Through correlation analysis, the results show that SNP19140160, SNP23355498, SNP9639603 and SNP9639605 molecular markers are obviously related to the body length of the F1 generation of the largehead jewfish stock, and the SNP19140160 locus is obviously related to the body weight, height, body thickness and body length of the F1 generation of the largehead jewfish stock. Through further analysis of the relation between genotypes and growth traits of the 4 SNP molecular markers, when the genotype of the SNP19140160 locus is AA, the weight, the body length, the height and the body thickness can be obviously improved; when genotypes of SNP9639603 locus are GG and TT, the body length is obviously higher than that of heterozygous GT; when the genotype of the SNP9639605 site is AA, the body length is obviously higher than that of heterozygous AT; when the SNP23355498 sites are AA and GG, the weight, the body length and the body height are all obviously higher than those of heterozygosity AG. Meanwhile, the molecular marker provided by the invention is verified by breeding work, is accurate and efficient, has very high operability and remarkable breeding effect, and can be used for breeding new varieties of quick-growth type large-mouth black bass new lines.
Drawings
FIG. 1 shows the result of the additive effect of weight in the F1 generation for 4 fast-growing markers.
Detailed Description
The invention provides a molecular marker for breeding quick-growth micropterus salmoides, which comprises at least one SNP molecular marker as follows; SNP19140160, SNP23355498, SNP9639603, and SNP9639605; the SNP19140160 is a site A/C with polymorphism at the position 19140160 of NW_024044570.1 chromosome in REFSEQ GCF _014851395.1 gene version; the SNP23355498 is a polymorphic site G/A at the position 23355498 of NW_024044348.1 in REFSEQ GCF _014851395.1 gene version; the SNP9639603 is the existence of a polymorphic site T/G at the position 9639603 of the NW_024044237.1 chromosome in REFSEQ GCF _014851395.1 gene version; the SNP9639605 is the presence of a polymorphic site T/A at position 9639605 of chromosome NW_024044237.1 in the REFSEQ GCF _014851395.1 gene version.
In the present invention, the SNP19140160 is a polymorphic site A/C at position 100 in the sequence shown in SEQ ID NO. 3. SNP23355498 is a polymorphic site G/A at position 100 in the sequence shown in SEQ ID NO. 13. SNP9639603 is a polymorphic site T/G located at position 100 in the sequence shown in SEQ ID NO. 10. SNP9639605 is a polymorphic site T/A located at position 100 in the sequence shown in SEQ ID NO. 12.
In the invention, simplified genome sequencing is carried out on the fast-growing (fast-growing) micropterus salmoides and the slow-growing (slow-growing) micropterus salmoides, high-quality sequences are extracted, the sequences are compared with a reference genome, SNP loci are detected, and the occurrence frequency of individuals with four genotypes (A, C, G and T) in fast-growing and slow-growing populations is counted. The threshold value of the frequency of the specific genotypes of the population is set to be 0.7, and for four genotypes, if the frequency of occurrence of individuals of the genotypes in the fast-growing population is greater than the threshold value of the frequency of the specific genotypes of the population, and the frequency of occurrence of individuals of the genotypes in the slow-growing population is less than 0.3, the genotypes are the specific genotypes of the fast-growing population. Wherein, the quick-growing micropterus salmoides preferably refers to a group of groups with the growth speed being 15% faster than the average value of the group growth speed. The slow-growing micropterus salmoides preferably refer to a group of populations with growth rates 15% slower than the average of the population growth rates.
13 Fast-growing SNP loci are obtained through screening by the method, and through correlation analysis of molecular markers and growth traits, SNP19140160, SNP23355498, SNP9639603 and SNP9639605 loci are obviously related to the body length of the F1 generation of the stock of the micropterus salmoides measured in experiments; the SNP19140160 locus is obviously related to the weight, height, thickness and length of the F1 generation of the largemouth black bass stock. When the genotype of the SNP19140160 site is AA, the weight, the body length, the height and the body thickness can be obviously improved. When genotypes at the 9639603 locus of SNP are GG and TT, the body length is significantly higher than that of heterozygous GT. When the genotype of the SNP9639605 site is AA, the body length is significantly higher than that of heterozygous AT. When the SNP23355498 sites are AA and GG, the weight, the body length and the body height are all obviously higher than those of heterozygosity AG.
The invention provides a primer for detecting quick-growing micropterus salmoides, which comprises a primer pair for amplifying the molecular markers; the primer pair for amplifying SNP19140160 molecular markers comprises a forward primer with a nucleotide sequence shown as SEQ ID NO. 18 and a reverse primer with a nucleotide sequence shown as SEQ ID NO. 19; the primer pair for amplifying SNP23355498 molecular markers comprises a forward primer with a nucleotide sequence shown as SEQ ID NO. 36 and a reverse primer with a nucleotide sequence shown as SEQ ID NO. 37; the primer pair for amplifying SNP9639603 molecular markers comprises a forward primer with a nucleotide sequence shown as SEQ ID NO. 32 and a reverse primer with a nucleotide sequence shown as SEQ ID NO. 33; the primer pair for amplifying SNP9639605 molecular markers comprises a forward primer with a nucleotide sequence shown as SEQ ID NO. 34 and a reverse primer with a nucleotide sequence shown as SEQ ID NO. 35.
The invention provides a kit for detecting rapid growth type micropterus salmoides, which comprises the primer and a PCR reaction solution. The PCR reaction solution is not particularly limited, and a solution for PCR amplification known in the art may be used.
The invention provides application of the molecular marker, the primer pair or the kit in auxiliary breeding of a fast-growing micropterus salmoides variety.
In the invention, the primer is used for amplifying the molecular marker of the sample to be bred, and when the genome DNA of the sample to be bred enriches the dominant growth genotype of the molecular marker and the growth speed of the sample to be bred is improved by more than 12% compared with that of the non-bred population, the sample to be bred is used as a material for breeding.
In the invention, the method for amplifying the molecular marker of the sample to be detected by using the primer preferably adopts a multiplex PCR amplification method; the amplification system for one round of PCR was 10. Mu.l, which included 3.1. Mu.l ddH 2 O, 1. Mu.l 10 XBuffer, 2. Mu.l of 50nM each of the upstream and downstream primers (each SNP site amplification primer), 0.8. Mu.l of 2.5mM dNTPs, 5U/. Mu.l Taq enzyme 0.1.8. Mu. l l, 2.8. Mu.l of template, and 1.8. Mu.l of 100mM Mg 2+. The reaction procedure for one round of PCR is preferably 95℃for 15min;94 ℃ for 30s,60 ℃ for 10min and 72 ℃ for 30s;4 cycles; 94℃30s,60℃1min,72℃30s,20 cycles. The reaction system for the two-round PCR amplification is preferably 20. Mu.l, including 3.5. Mu.l ddH 2 O, 2ul 10 XBuffer, 2uM Barcode 3.6. Mu.l, 2.5mM dNTP 0.8. Mu.l, 5U/. Mu.l Taq enzyme 0.1. Mu.l, 10. Mu.l of 10-fold diluted one-round PCR amplification product, 100mM Mg 2+ 1.0.0. Mu.l. The reaction procedure of the two-round PCR is preferably 95 ℃ for 15min;94 ℃ for 30s,60 ℃ for 4min and 72 ℃ for 30s;5 cycles; 94℃30s,65℃1min,72℃30s,10 cycles. Two rounds of PCR different samples were distinguished by different Barcode primers (Barcode contains the sample tag sequence). The barcode is an "identity card" of a mixed sample in sequencing, used to distinguish between different samples. In an embodiment of the present invention, the barcode includes ATCACG, CGATGT, TTAGGC, TGACCA, ACAGTG, GCCAAT, CAGATC, ACTTGA, GATCAG, TAGCTT, GGCTAC, CTTGTA.
And determining the sequence of the amplified product by adopting a second generation sequencing method after the amplified product is obtained, and judging the result of the sample to be detected according to the specific genotype of the molecular marker in the amplified product.
In the present invention, the dominant growth genotypes of the molecular markers preferably include 2 or more of the following SNP molecular marker dominant growth genotypes: the dominant growth genotype of SNP19140160 is AA; the dominant growth genotype of SNP9639603 is GG or TT; the dominant growth genotype of SNP9639605 is AA; the dominant growth genotype of SNP23355498 is AA or GG.
In the invention, the method is adopted to judge the growth type of the micropterus salmoides, the rapid growth type micropterus salmoides is selected for subsequent breeding, and experiments show that the breeding method can rapidly realize the aggregation of a plurality of rapid growth markers, effectively improve the growth speed of the offspring of breeding and obviously improve the growth speed of F3 generation compared with the group without breeding on the premise of ensuring genetic distance and genetic diversity.
The molecular markers for breeding the rapid-growth micropterus salmoides and the application thereof provided by the invention are described in detail below by combining with the examples, but the molecular markers are not to be construed as limiting the protection scope of the invention.
Example 1
Screening method for identifying molecular markers of quick-acting long largemouth black bass strain
1. Population selection and trait measurement
And selecting an F1 generation parent of the American stock of the Japanese black bass with 2000 reserved seeds, wherein each individual carries PIT marks. The markers are scanned separately and the body weight, length, height and body thickness index of each individual is measured. The parent with the weight of 1.5% of the ranking is selected as a parent of the F1 generation of the fast-growing largehead perch stock (30 samples), the parent with the weight of 1.5% of the ranking is selected as a parent of the F1 generation of the slow-growing largehead perch stock (30 samples), and the tail fin samples are respectively cut and stored in absolute ethyl alcohol for later use.
2. Simplified genomic sequencing of fast and slow-growing largemouth jewfish U.S. stock
The tail fin DNA from the fast and slow 30 samples were extracted separately and sent to the seaperson biotechnology, inc. Full genomic DNA was digested with restriction enzyme HindIII_MseI, cut fragments of a length were recovered and inserted, pooled according to double digestrestriction site-associatedDNAsequencing (dd-RAD), and the library was double-ended (Paired-end, PE) sequenced using the second generation sequencing technique (Next-Generation Sequencing, NGS) based on the IlluminaNovaSeq sequencing platform. Sequencing data contains some reads with joints and low quality, which can cause great interference to subsequent information analysis, and in order to ensure the quality of the subsequent information analysis, the next machine data needs to be further filtered, namely the original next machine data (rawdata) is filtered to generate a high-quality sequence (high qualitydata). The raw data was filtered using fastp (v0.20.0) using a sliding window method. The genome sequencing and data analysis process was completed by the Sharpeno biosciences, inc. of Sharpedo.
3. Screening of fast-growing SNP markers
The high quality data obtained after filtration was aligned to the reference genome using the bwa (0.7.12-r 1039) mem program (https:// www.ncbi.nlm.nih.gov/genome/. SNP (single nucleotide polymorphism) mainly refers to a polymorphism of a DNA sequence at a genomic level caused by variation of a single nucleotide, including single base transition, transversion, etc. SNP detection was performed using GATK software. The frequency of occurrence of individuals of the four genotypes in the fast-growing and slow-growing populations is counted respectively. The threshold value of the frequency of the population-specific genotypes is set to be 0.7, and for four genotypes (A, C, G, T), if the frequency of occurrence of individuals of the genotype in the fast-growing population is > the threshold value of the frequency of occurrence of the population-specific genotypes, and the frequency of occurrence of individuals of the genotype in the slow-growing population is <0.3, the genotype is the specific genotype of the fast-growing population. The specific calculation is as follows: genotypes are classified into three types of 0/0, 0/1 and 1/1 (if not detected, marked as deletion), the occurrence frequency of each genotype in the A group and the B group is counted respectively, and if the A group specific marker is found, the frequency is higher than 0.8 (0.6) in the A group at a certain site, and the frequency of the site is lower than (1-0.8) or (1-0.6) in the B group, the site is specific to the A group. And the specific case of group B is the reverse. 13 SNP loci which can be used for screening the rapid growth type micropterus salmoides are screened according to the standard, and the specific sequence and position information are shown in Table 1.
Table 1 information of screening 13 SNP molecular markers
Note that: the above fragment was designated as a genome version number REFSEQGCF _014851395.1.
4. Correlation analysis of fast-growing SNP markers and growth traits
Corresponding rapid-growing population-specific SNP site primers were designed (see Table 2). Randomly selecting 230 generations of U.S. stock F1 of micropterus salmoides cultured in the same pond, wherein each individual carries PIT marks. And respectively scanning marks, and measuring indexes such as weight, body length, height, body thickness and the like of each individual. And cutting a tail fin sample after each tail is detected, and storing the tail fin sample in absolute ethyl alcohol for later use. And carrying out second generation sequencing of the sites by using corresponding SNP specific primers, introducing SNP genotyping and growth traits into TASSEL software, carrying out correlation analysis by using a general linear model GLM, and carrying out growth trait correlation positioning. If the level of significance P <0.05, the locus is considered to be significantly associated with the trait. Screening positive SNP loci for subsequent molecular marker assisted breeding.
TABLE 2 design of SNP site primers specific to fast growing populations
The results are shown in Table 3.
TABLE 3 SNP loci that are significantly correlated with growth traits
Note that: p_marker is a significance level P value; rsq _marker is the marked pair type variant interpretation rate, represents the contribution size, and is considered as the main effect QTL if R 2 is more than 5%.
Correlation analysis shows that SNP19140160, SNP23355498, SNP9639603 and SNP9639605 sites are obviously related to the body length of the F1 generation of the stock of the micropterus salmoides measured in experiments; the SNP19140160 locus is obviously related to the weight, the height, the body thickness and the body length of the F1 generation of the stock of the largemouth black bass, wherein the contribution R 2 to the weight, the height and the body thickness is more than 5 percent, and the main effect QTL can be realized.
Grouping each character phenotype value according to genotypes of 4 associated SNPs, taking the grouping as a factor, taking the character phenotype value as a variable, performing single-factor analysis of variance by using SPSS 22.0 software, and outputting an average value and a standard deviation of each genotype grouping. The pairwise comparison method selects LSD statistical analysis. The results are shown in Table 4.
Table 44 correlation of SNP locus genotypes and measurement index
Note that: different lower case letters represent significant differences.
The result shows that when the genotype of the SNP19140160 locus is AA, the weight, the body length, the body height and the body thickness can be obviously improved. When genotypes at the 9639603 locus of SNP are GG and TT, the body length is significantly higher than that of heterozygous GT. When the genotype of the SNP9639605 site is AA, the body length is significantly higher than that of heterozygous AT. When the SNP23355498 sites are AA and GG, the weight, the body length and the body height are all obviously higher than those of heterozygosity AG.
The accumulated number of the associated markers is correlated with the characters, samples are grouped according to the number of the individual enriched dominant genotypes, the inter-group variance analysis is carried out by using SPSS 22.0 software, and the multiple comparison analysis is carried out by using an LSD method. The properties of each packet data were imported into graphpadprism5.0 software for mapping.
Analysis of the additive effect results (FIG. 1) showed that when 3 SNP sites were enriched, the average body weight was 647.13g, which was 12.21% higher than when two SNP marker sites were enriched (576.73 g). However, when 4 SNP sites were enriched, the average body weight was 703.86g, which was increased by 8.76% over that when 3 marker sites were enriched (647.13 g), and 22.04% over that when 2 marker sites were enriched. Therefore, the enrichment proportion of 4 sites of the breeding offspring is increased, and the growth performance can be effectively improved. Wherein in the randomly selected 230-tailed F1 generation population, the 4 site-enriched individuals have 35 tails, accounting for 15.2%.
Meanwhile, according to polymorphism detection results of other sequencing development sites besides the 4 screened rapid-growth SNP sites, the sites with the polymorphism information content higher than 0.25 are screened. When the content of the polymorphism information is more than 0.25, the loci are moderately polymorphic and the polymorphism is rich, so that the SNP marker loci have higher effectiveness and reliability when being used for genetic diversity analysis. The positions of the screened 27 polymorphisms are shown in Table 5.
TABLE 5 SNP loci screened for high polymorphism
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Genotype information of the 27 SNP/Indel markers of the 4 fast-growing marker enriched populations is imported into Powermarker software, allele frequencies are calculated, genetic distances based on the allele frequencies are calculated according to a method of Nei report (Nei,M.(1983)Genetic Polymorphism and the role of mutation in evolution.In:Nei,M.and Kohen,R.K.Eds.,Evolution of Gene and Proteins,Sinaver Associates,Sunderlans,165-190.), a genetic distance matrix is derived, and genetic distances among different individuals are compared.
5. Parental grouping and breeding
The F1 generation group firstly screens individuals with weight ranking of top 60 percent, complete body type, tidy tail lines and no deformity according to a group subculture breeding method. Cutting a tail fin sample, extracting a DNA sample, performing PCR amplification and second generation sequencing by using the screened specific primers of the positive SNP locus, detecting the genotype and genetic distance of the SNP marker locus of the selected parent, and selecting parent fishes for matched breeding by taking the principle of increasing the enrichment and homozygosity of the rapid-growth molecular markers of offspring and improving the genetic diversity of offspring groups to obtain F2 generation. And (3) selecting a matching principle: the parent contains 4 positive SNP loci, and the genetic distance between the paired parents is more than 0.15.
6. Continuous multiple generation breeding
From 10 ten thousand F2 offspring seeds, 685 offspring are selected as breeding parent fish of the next generation, and F2 parent is matched according to the detection method of F1 generation. Selecting, detecting and matching F3 generation parent according to the same method, selecting 637 tails from 10 ten thousand F3 generation fries as next generation breeding parent fish, and breeding F4 generation. And through 3-generation continuous breeding, a new strain of the quick-growing micropterus salmoides is cultivated.
Example 2
The relationship between the sex of the micropterus salmoides was verified based on the 4 rapid-growth SNP markers screened in example 1
A number of male and female larch were selected, and the body weight of each fish was weighed, and the genotypes of the 4 rapid-growth SNP markers in each individual were detected as in example 1. Based on the relation of genotypes corresponding to the SNPs of the micropterus salmoides with different sexes detected by the independent samples T, whether 4 rapid-growth SNPs are related to the sex difference of the micropterus salmoides or not is analyzed.
TABLE 6 sex relationship of quick-growth markers and largemouth black bass
| Index (I) | Male fish | Female fish |
| Quantity (tail) | 15 | 20 |
| Body weight (g) | 682.40±47.86 | 719.95±46.15 |
The result shows that 4 rapid-growth SNP markers have polymerized individuals in the male and female fishes, and meanwhile, the weights of the male and female fishes are not significantly different (P > 0.05), so that the selected 4 rapid-growth SNP markers have no sex linkage relationship and can be used as molecular markers for breeding of rapid-growth new lines.
Example 3
Auxiliary breeding screening of novel fast-growing lines of micropterus salmoides by using 4 fast-growing SNP markers screened in example 1
Genetic diversity and genetic distance analysis were performed for different generations based on genotype information of 27 SNP/Indel markers. Meanwhile, the enrichment rates of the 4 fast-growing marker enriched populations in different generations are calculated according to the formula I.
Enrichment (%) = (number of 4 fast-growing markers enriched individuals/total number of samples) ×100 equation I
TABLE 7 enrichment and genetic differentiation of quick growth markers for different breeding generations
| F1 | F2 | F3 | |
| Enrichment of 4 fast growth markers (%) | 15.2c | 57.5b | 89.6a |
| Genetic diversity | 0.348 | 0.337 | 0.302 |
| Genetic distance | 0.371 | 0.306 |
In the F2 generation population, 4 site-enriched individuals account for 57.5%; in the F3 generation group, the number of individuals enriched in 4 sites accounts for 89.6%, and the number of individuals enriched in the fast growth markers is obviously increased along with the increase of the breeding generation. The genetic diversity of the continuous 3-generation breeding population is reduced from 0.348 to 0.302. Polymorphism Information Content (PIC) is an indicator of the measure of gene polymorphism. A low polymorphic site when PIC < 0.25; at 0.25< pic <0.5, a moderate polymorphic site; PIC >0.5 indicates that the site is a highly polymorphic site. After 3 generations of breeding, the genetic diversity of the F3 generation still maintains the moderate polymorphic site. The genetic distance between the breeding generations is 0.306-0.371, the genetic distance between F1 and F2 is 0.371, the genetic distance between F2 and F3 is 0.306, and the genetic distances between adjacent generations are gradually reduced. The method shows that the genetic background of the breeding population tends to be consistent with the promotion of the breeding time, and the genetic structure also tends to be stable gradually.
Example 4
Selecting 9 culture ponds of 2 mu in Jiangsu without tin, wherein 3 ponds are used as breeding groups, each pond is respectively bred with 4000 new strains of the micropterus salmoides bred for 3 generations in the example 3, and the specification of fish bodies is about 30 g; putting the ungreeded largehead jewfish into 3 ponds as an ungreeded group, and putting 4000 ponds into each pond, wherein the specification is about 30 g; meanwhile, 4000 micropterus salmoides were purchased from other fine variety fields with a specification of about 30g as a control group. The breeding group, the non-breeding group and the control group are respectively fed with the same feed (the puffed compound feed 8902 for the weever of the biological technology limited company of Wuxi Tongwei, the protein 46 percent and the fat 8 percent). Morning 7: feeding 1 time at 00 pm and 17:00 pm respectively, wherein the feeding amount is 3% -4% of the weight of the fish. The cultivation period is 150 days. The test results are shown in Table 8.
TABLE 8 comparison of body weights of the selected and unselected groups and the control group micropterus salmoides
| Index (I) | Breeding group | Non-breeding group | Control group |
| Average initial body weight (g) | 29.46±3.28 | 30.38±3.63 | 30.12±3.29 |
| Average terminal body weight (g) | 534.29a±46.77 | 469.38b±41.93 | 451.26b±42.17 |
Note that: different letters represent significant differences in the same group (P < 0.05).
The body weight of the breeding group is improved by 13.8 percent and 18.4 percent compared with that of the non-breeding group and the control group, and the growth speed of the breeding group is obviously improved.
Therefore, by utilizing the molecular marker assisted breeding technology for breeding the novel strain of the quick-growth micropterus salmoides, the aggregation of a plurality of quick-growth markers can be realized quickly, the growth speed of the breeding offspring can be improved effectively on the premise of ensuring the genetic distance and the genetic diversity, and the growth speed of the F3 generation is improved by 13.8 percent compared with that of the non-breeding group.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Claims (4)
1. The molecular marker for breeding the rapid growth type micropterus salmoides is characterized by comprising the following SNP molecular markers: SNP19140160, SNP23355498, SNP9639603, and SNP9639605;
The SNP19140160 is a site A/C with polymorphism at the position 19140160 of NW_024044570.1 chromosome in REFSEQ GCF _014851395.1 gene version;
the SNP23355498 is a polymorphic site G/A at the position 23355498 of NW_024044348.1 in REFSEQ GCF _014851395.1 gene version;
The SNP9639603 is the existence of a polymorphic site T/G at the position 9639603 of the NW_024044237.1 chromosome in REFSEQ GCF _014851395.1 gene version;
The SNP9639605 is the existence of a polymorphic site T/A at the position 9639605 of chromosome NW_024044237.1 in REFSEQ GCF _014851395.1 gene version;
the SNP19140160 is a nucleotide sequence shown as SEQ ID NO. 3, and the polymorphism site A/C of the 100 th site in the nucleotide sequence shown as SEQ ID NO. 3;
the SNP23355498 is a nucleotide sequence shown as SEQ ID NO. 13, and the polymorphism site G/A of the 100 th site in the nucleotide sequence shown as SEQ ID NO. 13;
The SNP9639603 is a nucleotide sequence shown as SEQ ID NO.10, and the polymorphism site T/G of the 100 th site in the nucleotide sequence shown as SEQ ID NO. 10;
SNP9639605 is a polymorphic site T/A located at position 100 in the sequence shown in SEQ ID NO. 12.
2. A primer for detecting a rapid-growth micropterus salmoides, which is characterized by comprising a primer pair for amplifying the molecular marker of claim 1;
the primer pair for amplifying SNP19140160 molecular markers comprises a forward primer with a nucleotide sequence shown as SEQ ID NO. 18 and a reverse primer with a nucleotide sequence shown as SEQ ID NO. 19;
the primer pair for amplifying SNP23355498 molecular markers comprises a forward primer with a nucleotide sequence shown as SEQ ID NO. 36 and a reverse primer with a nucleotide sequence shown as SEQ ID NO. 37;
The primer pair for amplifying SNP9639603 molecular markers comprises a forward primer with a nucleotide sequence shown as SEQ ID NO. 32 and a reverse primer with a nucleotide sequence shown as SEQ ID NO. 33;
The primer pair for amplifying SNP9639605 molecular markers comprises a forward primer with a nucleotide sequence shown as SEQ ID NO. 34 and a reverse primer with a nucleotide sequence shown as SEQ ID NO. 35.
3. A kit for detecting a rapid-growth micropterus salmoides, comprising the primer of claim 2 and a PCR reaction solution.
4. Use of the molecular marker of claim 1, the primer of claim 2 or the kit of claim 3 in the assisted breeding of a rapidly growing micropterus salmoides variety;
the auxiliary breeding method of the rapid growth type largemouth bass variety comprises the following steps:
Amplifying a molecular marker of a sample to be bred by using the primer of claim 2, wherein when the genome DNA of the sample to be bred enriches the dominant growth genotype of the molecular marker and the growth speed of the sample to be bred is improved by more than 12% compared with that of a non-bred population, the sample to be bred is used as a material for breeding;
The dominant growth genotypes of the molecular markers comprise more than 2 types of the following SNP molecular marker dominant growth genotypes:
the dominant growth genotype of SNP19140160 is AA;
the dominant growth genotype of SNP9639603 is GG or TT;
the dominant growth genotype of SNP9639605 is AA;
the dominant growth genotype of SNP23355498 is AA or GG.
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| CN107022647A (en) * | 2017-06-22 | 2017-08-08 | 中国水产科学研究院珠江水产研究所 | A kind of SNP marker related to Micropterus salmoides growth traits and its application |
| CN114134237A (en) * | 2021-12-29 | 2022-03-04 | 南京宁渔种业研究院有限公司 | SRAP molecular marker for identifying northern American subspecies and Youtipe No. 3 parents of micropterus salmoides and application thereof |
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| CN104073486A (en) * | 2014-06-23 | 2014-10-01 | 中国水产科学研究院珠江水产研究所 | SNP site related to rapid growth of largemouth black bass as well as identification method and application thereof |
| CN107022647A (en) * | 2017-06-22 | 2017-08-08 | 中国水产科学研究院珠江水产研究所 | A kind of SNP marker related to Micropterus salmoides growth traits and its application |
| CN114134237A (en) * | 2021-12-29 | 2022-03-04 | 南京宁渔种业研究院有限公司 | SRAP molecular marker for identifying northern American subspecies and Youtipe No. 3 parents of micropterus salmoides and application thereof |
Non-Patent Citations (3)
| Title |
|---|
| High-Density Linkage Map and Mapping for Sex and Growth-Related Traits of Largemouth Bass ( Micropterus salmoides);Chuanju Dong等;Frontiers in Genetics;20191010;第10卷;第960-971页 * |
| 基于微卫星标记和线粒体D-loop序列的5个大口黑鲈群体遗传变异分析;张帝等;水生生物学报;20220930;第29卷(第9期);第1277-1289页 * |
| 大口黑鲈转录组SNPs筛选及其与生长的关联分析;全迎春等;水生生物学报;20161121;第40卷(第6期);第1128-1134页 * |
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