CN113151487A - Molecular identification marker primer combination for quantitative character of stichopus japonicus and thorn and application method thereof - Google Patents
Molecular identification marker primer combination for quantitative character of stichopus japonicus and thorn and application method thereof Download PDFInfo
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- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
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- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/124—Animal traits, i.e. production traits, including athletic performance or the like
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- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/156—Polymorphic or mutational markers
Abstract
The invention relates to a molecular identification marker primer combination for the quantitative character of stichopus japonicus and thorn and an application method thereof, belonging to the field of molecular biology, wherein the primer combination is shown as SEQ IP NO:1-15, and the method specifically comprises the following steps: extracting DNA of a stichopus japonicus sample by adopting an SDS method, carrying out SNP genotyping on the sample to be detected by using a KASP combined marker, and detecting a special marker locus of the stichopus japonicus with more or less thorns; the molecular identification standard of the stichopus japonicus is as follows: each individual has at least 3 more multi-spine markers covered. The method can judge the thorn number of the stichopus japonicus from the genetic point of view.
Description
Technical Field
The invention belongs to the field of molecular biology, and particularly relates to a molecular identification marker primer combination for the number of thorns of stichopus japonicus (A. japonica) and an application method thereof.
Background
The molecular marker refers to heritable and detectable DNA sequence or protein, and the molecular marker refers to specific DNA fragment capable of reflecting certain difference in genome among organism individuals or populations. Compared with the morphological marker, biochemical marker and cytological marker used in traditional breeding, the DNA molecular marker has remarkable superiority and is not influenced by the development and environment of organism. The molecular marker assisted breeding is characterized in that the molecular marker is closely linked with a gene determining the target character, and the existence of the target gene can be known by detecting the molecular marker, so that the aim of selecting the target character is fulfilled. The molecular marker assisted breeding can be used as an auxiliary means for identifying parent genetic relationship, transferring quantitative characters and recessive characters in backcross breeding, selecting hybrid progeny, predicting hybrid vigor, identifying variety purity and other breeding links. The molecular marker assisted breeding has important significance in the genetic breeding of aquatic animals, and the character characteristics of the obtained offspring can be determined by detecting the molecular marker, so that the missing of target characters can be effectively avoided; meanwhile, the molecular marker information can also be used for analyzing the genetic variation level of filial generation to guide population breeding. Among them, the single nucleotide polymorphism markers have the characteristics of low cost and high efficiency, and play an important role in genetic breeding.
In the breeding technology of aquatic economic animals, individuals with different genetic traits are selected by utilizing molecular markers, and the method has guiding significance in parent selection before the breeding period. The molecular marker assisted breeding technology is fast and efficient, is suitable for aquatic animals with large egg laying and sperm discharging quantity, can be directly used in the production link of parent selection, and is widely applied to the fine breed cultivation of the aquatic animals. Stichopus japonicus is a common seafood in north, and the sensory evaluation standard of high-quality Stichopus japonicus can be summarized as 'black, glutinous, and thorny'. The important breeding characteristics of the state-examined new species stichopus japonicus named as 'Water institute No. 1' and stichopus japonicus named as 'Anyuan No. 1' are 6 rows of stichopus japonicus, and the number of the spines is more than 45, which is obviously higher than that of the common stichopus japonicus. The stichopus japonicus with the thorny character is popular in the market, and the cultivation of the stichopus japonicus can bring remarkable economic benefit. The thorn character is determined by genetic factors and is influenced by various environmental factors (for example, temperature is one of the factors, the thorn quantity of the stichopus japonicus in the same family is different in different temperature environments), and the selection of parents is carried out only by utilizing the thorn phenotype in the breeding process, so that the selection of multi-thorn parents is missed or mistakenly carried out. The molecular identification marker primer combination for the quantity character of the stichopus japonicus spines and the application method thereof select seeds from the gene perspective, judge and identify the quantity of the stichopus japonicus spines from the genetic perspective, can realize early selection of economic characters, and are more accurate in seed selection and more economic and economical.
Disclosure of Invention
The invention aims to solve the technical problem of providing a labeled primer combination for identifying the number of stichopus japonicus thorn and an application method thereof, wherein the labeled primer combination can judge the number of stichopus japonicus thorn from the genetic aspect.
The invention is realized by the following technical scheme:
a molecular identification marker primer combination for the quantitative character of stichopus japonicus thorn is shown in Table 1;
TABLE 1 KASP marker combinations for detecting the number of thorns in Stichopus japonicus
The shaded sites are the unique genotypes of the spiny sea cucumber.
The invention also provides a method for identifying the number of the stichopus japonicus wart feet by using the primer combination, which comprises the following steps: extracting DNA of the stichopus japonicus sample by adopting an SDS method, carrying out SNP genotyping on the sample to be detected by using KASP combined markers, and detecting the special marker sites of the stichopus japonicus with more or less thorns. The judgment standard is as follows: in the actual production, all individuals carrying the stichopus spinosus marker do not need to be detected, and in order to ensure the detection accuracy, the molecular identification standard of the stichopus spinosus is as follows: each individual has at least 3 markers coverage, and does not carry any spine-less markers.
Detailed Description
The technical solution of the present invention is further explained by the following examples, but the scope of the present invention is not limited in any way by the examples.
Example 1: screening for markers
The research uses the whole genome sequence of the stichopus japonicus as reference, 215 representative stichopus japonicus in 8 different regions are subjected to genome re-sequencing respectively, and SNP marker sites related to the number of wart feet of the stichopus japonicus are obtained by comparing the reference genome; therefore, KASP primer design and genotyping are carried out, and the stichopus japonicus group with different wart foot number measuring amounts is verified, and finally the SNP primer sequence combination for identifying the number of the wart feet of the stichopus japonicus is obtained. The specific method comprises the following steps:
1. mark acquisition
1.1DNA sample extraction
215 which are representative and have good state are selected, including Liaoning, Shandong, Russia (Iradev Stoke) and a new variety of stichopus japonicus named as Water institute No. 1 in the main production area of Liaoning stichopus japonicus in China. Wherein each 30 stichopus japonicus in the fields of tourism (yellow Bohai sea boundary), west mill (high temperature resistance), Shandong (low latitude), yellow dragon tail (yellow sea offshore), octopus island (yellow sea offshore), Pindao island (artificial culture) and Russia (Fraidivor Stoke) sea are extracted, 5 stichopus japonicus in No. 1 water yard is extracted by SDS method, and the DNA concentration is more than 100 ng/ul.
1.2 genome Re-sequencing
The procedures of sample quality detection, library construction, library quality detection, library sequencing and the like are all executed according to the protocol standard (second generation high throughput sequencing provided by Illumina). And randomly breaking the qualified DNA sample into fragments with the length of 350bp by a Covaris crusher, and performing end repair, adding a ployA tail, adding a sequencing linker, purifying and amplifying in a PCR instrument for 10 cycles to complete the preparation of the whole library. Performing genome re-sequencing on the library qualified by quality inspection on an Illumina platform, counting the number of sequences of original double-end sequence reads obtained by sequencing in a unit of every 4 lines, and performing quality evaluation, wherein the evaluation principle is as follows:
(1) removing the adapter;
(2) removing reads containing N (N represents that base information cannot be determined) with the proportion of more than 5%;
(3) removing low-quality reads (the base number with the quality value Q < -10 accounts for more than 20 percent of the whole reads);
(4) and counting the original sequencing quantity, the effective sequencing quantity, Q20, Q30 and GC content, and performing comprehensive evaluation.
The filtered Clean Reads were aligned to the reference genomic sequence in BWA software. And (3) carrying out SNP and InDel detection by utilizing GATK software according to the positioning result of clear Reads in a reference genome, carrying out quality filtration on the mutation sites according to the principle that MAF is greater than 0.05 and the data integrity is greater than 0.8, and reserving SNP sites with the two allelic gene polymorphisms.
1.3 Association analysis
Combining stichopus japonicus thorn data, performing association on SNP sites by adopting a GLM (generalized linear regression) model in plink software, calculating each SNP site to obtain an associated p value, and obtaining a significant threshold value through Bonferroni correction. And (3) drawing a Manhattan graph (detected by a QQ-plot graph) by adopting an emmax model, setting 0.95 and 0.99 as thresholds, and setting clustered sites above the thresholds as SNP sites related to the thorn quantitative character.
1.4 typing and screening of specific SNP sites
And summarizing all the different variation sites between the samples according to the comparison result of the samples and the reference genome, and screening 50 high-quality SNP sites for subsequent KASP verification. The screening principle is as follows: (1) the locus has different dominant genotypes in samples with different numbers of wart feet of the stichopus japonicus (for example, AA or AT genotypes are dominant in samples with few wart feet, and AA or AT genotypes are dominant in samples with more wart feet); (2) adjacent variant sites can not be too close, and the distance is more than 50 bp; (3) the upstream and downstream sequences of the site have uniqueness in the genome, and less repeated sequences exist near the site. Extracting the 200bp upstream and downstream sequences of the high-quality SNP locus to be developed into a marker.
1.5KASP primer design
50 pairs of PCR 3' end specific amplification primers are designed according to SNP sites, the design software is Primer 5, and each sequence needs 3 primers, namely two specific upstream primers and one universal downstream Primer. The principle of primer design is as follows: (1) the primers do not have hairpin structures, and primer dimers cannot be formed between the upstream primers and the downstream primers; (2) the GC content of the primer is 40-60%, and the Tm value is 55-65 ℃.
Verification of KASP tags
2.1 validation of population DNA extraction and marker detection
Randomly selecting 92 different batches of stichopus japonicus selenka (60 of which are marked source stichopus japonicus selenka population, and 30 of which are different batches of stichopus japonicus selenka)Ginseng, the number difference of wart feet is more than 10), extracting DNA by SDS method, and extracting DNA in Douglas Array
The method comprises the steps of carrying out label detection on Platform, carrying out fluorescent quantitative PCR reaction in a SOELLEX high-flux PCR water bath environment, and selecting TB Green as a reagentTM Premix Ex TaqTMII, preparing 20 mul mixed system to carry out specific amplification by a two-step reaction program of 'denaturation-annealing extension'.
2.2KASP marker typing
Signals of two fluorophores, namely FAM and VIC, are read by an ARAYA fluorescence reader, and the results are led into a database to carry out SNP typing on the sample on the Douglas Scientific Dashboard according to the principles of definite typing, NTC (no sample negative control) and no specific amplification. And analyzing the typing result by utilizing SNP viewer genotype reading software, and reading the heterozygous genotype by the KASP marker in the sample to obtain the successful typing. In the typing scheme, X: X represents homozygous type including five types A: A, T: T, C: C, G: G, - (deletion), and X: Y represents heterozygous type including three types A: G, C: T, A: - (partial deletion)? Representing no or weak signal, and uncalloble representing signal but no explicit typing. Finally, 10 KASP marks of successful typing are obtained.
The markers with the spiny character genotype as a heterozygote are removed from 50 KASP markers, and 5 markers with the highest detection rate are screened (see Table 1).
3. Identification and application of stichopus japonicus selenka
The obtained KASP markers were verified on 60 stichopus japonicus populations of the original batch (i.e., marker source population) and 32 stichopus japonicus grown in different batches, and the results showed that:
(1) for the original batch stichopus japonicus population (from 8 populations, 30 multi-stichopus japonicus and 30 few-stichopus japonicus, wherein the average number of thorns of the multi-stichopus japonicus is more than 45, the average number of thorns of the few-stichopus japonicus is less than 35); the mark coverage rate of the stichopus japonicus is 100%, and the average mark rate of each sample of the stichopus japonicus is 3.5.
(2) Selecting 32 stichopus japonicus populations of different batches, wherein (20 individual stichopus japonicus, the number of thorns is 48-65, 12 small stichopus japonicus, and 28-32 thorns), the mark coverage rate of the stichopus japonicus is 85%, the average mark rate of each sample of the stichopus japonicus is 2.5, and the marks are not mixed in the detection of the stichopus japonicus and the small stichopus japonicus.
(3) Examples of early selection: selecting 2 cultured stichopus japonicus populations, randomly selecting 30 stichopus japonicus populations for cultivation at 0.5 age, and respectively obtaining the average thorn number of 36 and 28 when the stichopus japonicus populations are 2 ages (capable of reproducing offspring). Meanwhile, when the 2 stichopus japonicus populations are 0.5-year old, the labeling detection is carried out, 30 stichopus japonicus containing at least 3 multi-spine markers and not carrying few spine markers are screened out for culture, and when the 2-year-old stichopus japonicus population (capable of being bred into offspring), the average number of spines is 62 and 46 respectively.
(4) The method can be applied to selective breeding of the stichopus japonicus, and the stichopus japonicus seeds carrying at least 3 multi-thorn markers (not carrying few-thorn markers) are selected by utilizing the markers according to the method for subsequent propagation and breeding, so that a new variety of the stichopus japonicus can be cultivated.
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Claims (2)
1. A molecular identification marker primer combination for the quantitative trait of stichopus japonicus thorn is characterized in that the marker primer combination is shown in the following table 1:
TABLE 1 KASP marker combinations for detecting the number of thorns in Stichopus japonicus
Wherein, the shadow marked sites are the special genotypes of the stichopus japonicus selenka.
2. The method for carrying out molecular identification on the quantity character of stichopus japonicus thorn by using the primer combination of claim 1, which is characterized by comprising the following steps: extracting DNA of a stichopus japonicus sample by adopting an SDS method, carrying out SNP genotyping on the sample to be detected by using a KASP combined marker, and detecting a special marker locus of the stichopus japonicus with more or less thorns; the molecular identification standard of the stichopus japonicus is as follows: each individual has at least 3 markers coverage, and does not carry any spine-less markers.
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| CN202110175759.3A CN113151487A (en) | 2021-02-09 | 2021-02-09 | Molecular identification marker primer combination for quantitative character of stichopus japonicus and thorn and application method thereof |
| AU2021104302A AU2021104302A4 (en) | 2021-02-09 | 2021-07-19 | Marker primer combination for molecular identification of quantitative traits of spines of apostichopus japonicus and use thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114525362A (en) * | 2022-03-18 | 2022-05-24 | 大连海洋大学 | Primer combination for identifying Anyuan No. 1 stichopus japonicus population and application thereof |
| CN115948577A (en) * | 2023-01-09 | 2023-04-11 | 山东省渔业发展和资源养护总站 | Breeding method of high-temperature-resistant sea cucumbers |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113151487A (en) * | 2021-02-09 | 2021-07-23 | 大连海洋大学 | Molecular identification marker primer combination for quantitative character of stichopus japonicus and thorn and application method thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107937569A (en) * | 2018-01-16 | 2018-04-20 | 中国水产科学研究院黄海水产研究所 | A kind of molecular labeling and its application for stichopus japonicus growth traits assisted selection |
| CN112080570A (en) * | 2020-10-26 | 2020-12-15 | 大连海洋大学 | KASP labeled primer combination for identifying hybrid stichopus japonicus in Zhongrussia and application thereof |
| CN112251533A (en) * | 2020-11-09 | 2021-01-22 | 浙江省农业科学院 | SNP molecular marker closely linked with main effect QTL site of eggplant fruit length and application |
| AU2021104302A4 (en) * | 2021-02-09 | 2021-08-19 | Dalian Ocean University | Marker primer combination for molecular identification of quantitative traits of spines of apostichopus japonicus and use thereof |
-
2021
- 2021-02-09 CN CN202110175759.3A patent/CN113151487A/en active Pending
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107937569A (en) * | 2018-01-16 | 2018-04-20 | 中国水产科学研究院黄海水产研究所 | A kind of molecular labeling and its application for stichopus japonicus growth traits assisted selection |
| CN112080570A (en) * | 2020-10-26 | 2020-12-15 | 大连海洋大学 | KASP labeled primer combination for identifying hybrid stichopus japonicus in Zhongrussia and application thereof |
| CN112251533A (en) * | 2020-11-09 | 2021-01-22 | 浙江省农业科学院 | SNP molecular marker closely linked with main effect QTL site of eggplant fruit length and application |
| AU2021104302A4 (en) * | 2021-02-09 | 2021-08-19 | Dalian Ocean University | Marker primer combination for molecular identification of quantitative traits of spines of apostichopus japonicus and use thereof |
Non-Patent Citations (5)
| Title |
|---|
| GERARD DOWNEY编著: "《Advances in Food Authenticity Testing》", 12 August 2016, WOODHEAD PUBLISHING * |
| ZHAN Y等: "Comparative transcriptome analysis identifies genes associated with papilla development in the sea cucumber Apostichopus japonicus.", 《COMP BIOCHEM PHYSIOL PART D GENOMICS PROTEOMICS.》 * |
| 倪萍 等: "仿刺参疣足数量SNP遗传力评估", 《渔业科学进展》 * |
| 李云霞等: "基于微卫星标记的刺参群体遗传结构分析及与经济性状的相关性研究", 《大连海洋大学学报》 * |
| 郑龙华等: "刺参多刺品系子三代池塘养殖中试与效果评价", 《河北渔业》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114525362A (en) * | 2022-03-18 | 2022-05-24 | 大连海洋大学 | Primer combination for identifying Anyuan No. 1 stichopus japonicus population and application thereof |
| CN114525362B (en) * | 2022-03-18 | 2023-05-23 | 大连海洋大学 | Primer combination for identifying Apostichopus japonicus population of Anyuan No. 1 and application thereof |
| CN115948577A (en) * | 2023-01-09 | 2023-04-11 | 山东省渔业发展和资源养护总站 | Breeding method of high-temperature-resistant sea cucumbers |
| CN115948577B (en) * | 2023-01-09 | 2023-08-22 | 山东省渔业发展和资源养护总站 | Breeding method of high-temperature-resistant sea cucumber |
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| AU2021104302A4 (en) | 2021-08-19 |
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