CN110129455A - A kind of application growing relevant molecular labeling in litopenaeus vannamei genetic breeding - Google Patents
A kind of application growing relevant molecular labeling in litopenaeus vannamei genetic breeding Download PDFInfo
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Abstract
The invention belongs to fishery breeding technical fields, and in particular to a molecular labeling relevant to weight and its application in weight character breeding on litopenaeus vannamei LvMMD2 gene.By integrating the methods of whole-genome association and candidate gene association analysis, in the label (LvMMD2-132-C/T) that the gene coding region LvMMD2 screens to prawn weight in significant related (P=2.42 × 10‑8).The label is located at the position 132bp of SEQIDLvMMD2, is mutated for C/T type, and the TT genotype of the label is growth preponderant genotype.Label according to the present invention can be used as the hereditary and selection of growth traits related molecular marker auxiliary prawn growth traits.
Description
Technical field
The invention belongs to aquatic livestock breeding fields, and in particular on litopenaeus vannamei LvMMD2 gene one with weight phase
The molecular labeling of pass and its application in weight character breeding.
Background technique
Litopenaeus vannamei (Litopenaeus vannamei), also known as Penaeus Vannmei (white Pacific
Shrimp), it is one of most important aquaculture kind in the world, occupies the 76% of world's prawn total output, culturing area
Throughout world various regions.The Variety comprehensive of litopenaeus vannamei also China's prawn culturing is introduced for the kind 1988 from the U.S.
It is supported in China's large-scale farming since 1999 until annual output in 2013 up to 1,400,000 tons or so, accounts for China prawn in China
Grow 80% or more of total output.
With the continuous expansion of cultivation scale, demand of the aquaculture industry to litopenaeus vannamei elite germplasm is increasingly urgent to.?
When carrying out the cultivation of prawn elite germplasm, growth traits is both important measurement index, while being also most direct breeding objective.So
And for a long time, the genetic improvement of prawn depends on artificial selection and the traditional breeding method based on Quantitative Genetics, passes
The problems such as hereditary and selection of uniting is low with selection accuracy there is also breeding period length, it is difficult to meet the reality for accelerating genetic improvement progress
It is required that.Compared to conventional genetic breeding, molecular marking supplementary breeding has the spies such as breeding interval is short, accuracy and reliability is high
Point can obviously accelerate the hereditary and selection progress of important economical trait.'s.
Molecular marking supplementary breeding is that molecular labeling is auxiliary applied to carrying out during animals and plants correlated traits genetic improvement
A kind of technological means for helping selection, its essence is carry out target area to individual using the molecular labeling with target gene close linkage
Domain and full-length genome screening, obtain desired individual, to improve Breeding Efficiency and accuracy rate.Early period studies people in prawn
Member has navigated to some QTL sites relevant to growth by QTL localization method, such as in litopenaeus vannamei QTL positioning, also sends out
The relevant QTL site of growth (Andriantahina F, Liu X, Huang H.Genetic map construction is showed
and quantitative trait locus(QTL)detection of growth-related traits in
Litopenaeus vannamei for selective breeding applications.PLoS One,2013,8:
E75206), but it is limited by lower mark density, does not navigate to the gene or label directly related with growth.This hair
Bright to be intended to provide one kind and the significant relevant label of growth traits, which may be used as the auxiliary of Growth Trait in White Shrimp Litopenaeus vannamei
Breeding.
Summary of the invention
The object of the present invention is to provide a kind of molecular labelings relevant to Growth of Litopenaeus vannamei, and are applied to prawn and grow
The molecular marking supplementary breeding of fast kind.
To achieve the above object, the invention adopts a technical scheme as:
It a kind of molecular labeling relevant to growth on litopenaeus vannamei LvMMD2 gene and its is lost in growth traits
Pass the application in breeding, which is characterized in that (1) extract prawn and reserve seed for planting the DNA of parent；(2) following primer LvMMDF is used:
ACAATCGCTTCAACCAACCTG and LvMMDR:TCCTGGGCGTGATCTTCTTT expands parent material DNA；(3)
It is sequenced using the segment of LvMMDF primer pair amplifies, obtains sequence shown in SEQIDLvMMD2, and analyze it
The genotype of the SNP site (LvMMD2-132-C/T) of the position 132bp, if the site LvMMD2-132-C/T only has in sequencing peak figure
The peak C, then the idiotype is CC, bimodal if C and T, then the idiotype is CT, if the only peak T, the genes of individuals
Type is TT.(4) mating strategy is designed according to genotyping result, prepares the TT genotype individuals with growth vigor, assisting growth
The hereditary and selection of shape.Further, the application of relevant molecular labeling is grown to prawn, the TT genotype individuals speed of growth is most
Fastly, followed by CT type, and the speed of growth of TT type individual is most slow.
In prawn reproductive process, by SNP classifying method by after the label parting, TT type individual is selected to carry out breeding energy
Enough assisting growths, to improve prawn yield.
Advantages of the present invention is as follows: the relevant molecular labeling of growth that this method provides can be from molecular level to growth
Fast individual is selected, and is not limited, and can be selected in early days by external environment and Individual Size, improves the effect of selection
Rate and accuracy rate, and aquaculture cost can be reduced, shorten the culture-cycle.
Detailed description of the invention
Three kinds of the site Fig. 1 LvMMD2-132-C/T genotype average weight distribution map in mixed breeding group.
Specific embodiment
The acquisition of embodiment 1:LvMMD2-132-C/T label
(1) whole-genome association (GWAS) of Growth Trait in White Shrimp Litopenaeus vannamei:
Material for GWAS analysis is a population mixture being made of 13 family full-sibs.In July, 2015, in sea
The ocean Nan Guangtai breeding companies (Wenchang, Hainan) amount to 13 family full-sibs of building by orienting mating mode.Each family
It is transferred to 5m respectively after hatching the young2Pond individually cultivated.Period, to reduce breeding environment to the shadow of different familys
It rings, each family is cultivated using standardized breeding way as far as possible, makes salinity, temperature, young density, bait and inflation etc.
Condition is consistent as far as possible.In September, 2015 is transferred to 10m after random 50 tail prawn of picking mixing each in 13 familys2Water
Pond is cultivated under same environment, forms population mixture.200 tail shrimps are randomly selected, electronics day is used in mid-November, 2015
It is flat to measure its weight, and carry out the judgement of gender.
The DNA of all material is extracted using Plant Genome extracts kit (Tiangeng, Beijing), later using 2b-RAD's
Method to it is all individual respectively carry out full-length genome SNP parting (Wang S, Meyer E, McKay JK, Matz MV, 2b-RAD:
a simple and flexible method for genome-wide genotyping,Nature methods,2012,
9,808-810) it, using the SNP typing data of acquisition, is analyzed using the GWAS that GenABEL software carries out prawn weight, positioning life
Long correlation SNP marker (Aulchenko Y.S., Ripke S., Isaacs A., van Duijn C.M.GenABEL:an R
package for genome-wide association analysis.Bioinformatics.23:1294-6
(2007).)。
(2) association analysis of the LvMMD2 as growth correlation candidate gene
It is analyzed by GWAS and obtains a batch and the label of weight significant related (P < 0.01).Base is carried out by marking to these
Because of annotation, discovery is located in one of SNP and is located at monocyte to macrophage differentiation factor 2
(LvMMD2) gene includes subregion.Using the gene (LvMMD2) as growth correlation candidate gene, in the gene coding region
SNP detailed survey is carried out, and carries out the association analysis of candidate locus to the nonsynonymous mutation SNP of code area.Analysis is as the result is shown
The SNP site of the position 132bp of SEQIDLvMMD2 sequence is in extremely significant related (P=2.42 × 10 to prawn weight-8), the position
For point to the explanation rate of weight phenotypic variation up to 10.5%, the TT genotype of the label is preponderant genotype.
SEQ ID LvMMD2
ACAATCGCTTCAACCAACCTGCACTGGGAGCATACCCATCCTACAAGCTAGAGCACTCAGGCAAAGAT
GCATCACATGTCTCAGGTGCCCTGCCTTCAACTCCCCCCACTATCCCCACAGCTGAGGGGGTG[C/T]TCTCAGAT
GCTGAAGGAAGAATCAGGTAGGTGAGGACAGCATAGTAATGGATAAAGGCTGCCAGGACCACAAAGAGATGCCATAT
GGCATGTGCCAGAGGAATGCGGCCATCCAGCTTAAAGAAGATCACGCCCAGGA
Embodiment 2:LvMMD2-132-C/T marks the application in breeding.
(1) parting in the candidate site Parent LvMMD2-132-C/T
Candidate Parent is used as with 60 tail parent shrimps of Hainan ocean Guang Tai breeding companies (Wenchang, Hainan) in June, 2016,
After taking a pair of of swimmeret of every tail prawn to extract DNA, the target area sequence of each individual of following primer pair amplifies is utilized:
LvMMDF:ACAATCGCTTCAACCAACCTG
LvMMDR:TCCTGGGCGTGATCTTCTTT
Pcr amplification product carries out generation sequencing using ABI3730xl sequenator, and sequencing primer is LvMMDF primer, according to
Peak figure is sequenced, parting is carried out to LvMMD2-132-C/T, if the position only has 1 C base peak, genotyping result is CC type, if
An only T base peak, then genotyping result is TT type, if there are T base peak, genotyping result in the position existing C base peak again
For CT type.
(2) prepared by family's based material
According to above-mentioned genotyping result, it is homozygous as parent's preparation CC and TT type that CC type individual and TT type individual are chosen respectively
Family.In addition, being oriented mating preparation CT type family using CC type and TT type individual as Parent.Each family hatches children
5m is transferred to after body respectively2Pond individually cultivated, when family Material growth is to 1cm, each random picking 50 of family
Tail litopenaeus vannamei carries out being put into 10m2Pond mixed breeding.It selects 200 individuals after cultivation 4 months at random from mixed breeding group and carries out body
It is fixed to resurvey.
(3) LvMMD2-132-C/T marker assisted selection
Genotyping is carried out in the site LvMMD2-132-C/T to above-mentioned individual.In above-mentioned material, detect for CC type,
Three kinds of genotype individuals of CT type and TT type, corresponding average weight is respectively 8.77 ± 3.22g, 9.52 ± 3.21g and 12.48
± 2.37g (Fig. 1).Therefore it is selected by the TT genotype in being marked to LvMMD2-132-C/T, breeding group can be improved
The speed of growth of body.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Therefore
Done without departing from theon the basis of the spirit of the present invention it is some modify or improve, fall within the scope of the claimed invention.
Sequence table
<110>Institute of Oceanology of the Chinese Academy of Sciences
<120>a kind of application for growing relevant molecular labeling in litopenaeus vannamei genetic breeding
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 270
<212> DNA/RNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<400> 1
acaatcgctt caaccaacct gcactgggag catacccatc ctacaagcta gagcactcag 60
gcaaagatgc atcacatgtc tcaggtgccc tgccttcaac tccccccact atccccacag 120
ctgagggggt gytctcagat gctgaaggaa gaatcaggta ggtgaggaca gcatagtaat 180
ggataaaggc tgccaggacc acaaagagat gccatatggc atgtgccaga ggaatgcggc 240
catccagctt aaagaagatc acgcccagga 270
Claims (5)
1. a kind of application for growing relevant molecular labeling in litopenaeus vannamei genetic breeding, it is characterised in that:
The relevant molecular labeling of the growth is located on litopenaeus vannamei LvMMD2 gene, with SEQ ID LvMMD2
Shown in nucleotide sequence, it is C/T type SNP marker which, which is located at the position 132bp of SEQIDLvMMD2 sequence, the label
TT genotype is the genotype of growth vigor.
2. application described in accordance with the claim 1, it is characterised in that:
, will be after the label parting by SNP classifying method in prawn breeding process, selecting TT type individual reserve seed for planting can be auxiliary
The selection of growth traits is helped, to improve prawn yield.
3. application described in accordance with the claim 1, it is characterised in that:
In prawn breeding process, by SNP classifying method by after the label parting, according to genotyping result, the first is preferably protected
The homozygous material of TT is stayed to be reserved seed for planting and passed on；No the first selection Shi Cixuan male parent female parent is respectively that CT type mates with TT type,
Selecting male parent female parent when no the first and second of selection is that CT type mates with CT type, and post-coitum, which filters out, has growth in offspring
The TT genotype individuals of the advantage genetic stocks fast as growth.
4. according to application described in claim 2 or 3, it is characterised in that: SNP parting specific operation process,
(1) prawn is extracted to reserve seed for planting the DNA of parent；
(2) following primer LvMMDF:ACAATCGCTTCAACCAACCTG and LvMMDR:TCCTGGGCGTGATCTTCTTT is used
Expand parent material DNA；
(3) it is sequenced using the segment of LvMMDF primer pair amplifies, obtains sequence shown in SEQIDLvMMD2, and analyze it
The genotype of the SNP site (LvMMD2-132-C/T) of the position 132bp, if the site LvMMD2-132-C/T only has in sequencing peak figure
The peak C, then the idiotype is CC, bimodal if C and T, then the idiotype is CT, if the only peak T, the genes of individuals
Type is TT.
5. according to application described in claim 1,2,3 or 4, it is characterised in that: the relevant molecular labeling of the growth refer to
The molecular labeling of litopenaeus vannamei weight character heredity.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111850133A (en) * | 2020-05-28 | 2020-10-30 | 广东省农业科学院动物科学研究所 | Low-temperature-resistant associated SNP molecular marker of litopenaeus vannamei, detection primer and application of detection primer |
| CN111926020A (en) * | 2020-07-24 | 2020-11-13 | 中国科学院海洋研究所 | Two prawn growth related genes and application thereof in genetic breeding |
| CN114107523A (en) * | 2022-01-27 | 2022-03-01 | 广东省农业科学院动物科学研究所 | Snp molecular marker associated with growth traits of litopenaeus vannamei, detection primer and application of detection primer |
| CN117344033A (en) * | 2023-12-06 | 2024-01-05 | 中国水产科学研究院黄海水产研究所 | Molecular marker related to growth of litopenaeus vannamei and application thereof |
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| US20170175188A1 (en) * | 2015-05-14 | 2017-06-22 | Institute Of Oceanology, Chinese Academy Of Sciences | Dna probe sequence for genetic sex identification of litopenaeus vannamei and acquisition method |
| CN109055580A (en) * | 2018-11-01 | 2018-12-21 | 中国科学院海洋研究所 | Litopenaeus vannamei c-type scavenger receptor is interior and grows related molecular marker and application |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111850133A (en) * | 2020-05-28 | 2020-10-30 | 广东省农业科学院动物科学研究所 | Low-temperature-resistant associated SNP molecular marker of litopenaeus vannamei, detection primer and application of detection primer |
| WO2021159661A1 (en) * | 2020-05-28 | 2021-08-19 | 广东省农业科学院动物科学研究所 | Low temperature resistance-related snp molecular marker of litopenaeus vannamei, and detection primer and use thereof |
| CN111850133B (en) * | 2020-05-28 | 2021-10-19 | 广东省农业科学院动物科学研究所 | Low-temperature-resistant associated SNP molecular marker of litopenaeus vannamei, detection primer and application of detection primer |
| CN111926020A (en) * | 2020-07-24 | 2020-11-13 | 中国科学院海洋研究所 | Two prawn growth related genes and application thereof in genetic breeding |
| CN114107523A (en) * | 2022-01-27 | 2022-03-01 | 广东省农业科学院动物科学研究所 | Snp molecular marker associated with growth traits of litopenaeus vannamei, detection primer and application of detection primer |
| CN114107523B (en) * | 2022-01-27 | 2022-05-13 | 广东省农业科学院动物科学研究所 | Snp molecular marker associated with growth traits of litopenaeus vannamei, detection primer and application of detection primer |
| CN117344033A (en) * | 2023-12-06 | 2024-01-05 | 中国水产科学研究院黄海水产研究所 | Molecular marker related to growth of litopenaeus vannamei and application thereof |
| CN117344033B (en) * | 2023-12-06 | 2024-03-19 | 中国水产科学研究院黄海水产研究所 | Molecular marker related to growth of litopenaeus vannamei and application thereof |
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