CN106577425A - Panmixis parent tracing method for litopenaeus vannamei - Google Patents
Panmixis parent tracing method for litopenaeus vannamei Download PDFInfo
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- CN106577425A CN106577425A CN201611201011.1A CN201611201011A CN106577425A CN 106577425 A CN106577425 A CN 106577425A CN 201611201011 A CN201611201011 A CN 201611201011A CN 106577425 A CN106577425 A CN 106577425A
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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Abstract
The invention discloses a panmixis parent tracing method for litopenaeus vannamei. The purposes of panmixis of litopenaeus vannamei and post parent tracing and identification are achieved in combination with a multiplication characteristic that spermatophore is still remained in the body of female litopenaeus vannamei after the litopenaeus vannamei spawns, and a litopenaeus vannamei microsatellite paternity test method which is mature currently. According to the method provided by the invention, the advantage that the female litopenaeus vannamei is high in successful mating efficiency during a production process is used, and the deficiency that the male does not mate with the female in directional mating and the position of the spermatophore is incorrect after mating is overcome; application of the method provided by the invention quickens the preparation of the full-sib families of the litopenaeus vannamei, and is beneficial to prompting the operation of litopenaeus vannamei molecular breeding.
Description
Technical field
The invention belongs to genetic breeding field, and in particular to a kind of Litopenaeus vannamei random mating parent's retroactive method.
Background technology
Litopenaeus vannamei (Litopenaeus vannamei, also known as Penaeus vannamei) aquaculture is in China Fisheries economy
Occupy larger proportion, be one of pillar industry of China's aquaculture.However, the genetic breeding work of China's Litopenaeus vannamei
But fall behind very much, the close shrimp of high-quality relies on import, healthy, stable and sustainable development of this situation to China's prawn industry for a long time
Exhibition brings larger hidden danger.Therefore, the basis and applied research for strengthening Litopenaeus vannamei genetic breeding field works, and will help
In the unfavorable situation for improving China's Litopenaeus vannamei industry.
It is indispensable link in molecular breeding work to prepare the retrospective family full-sibs of a large amount of parents.In Fish
In breeding work, can adopt in vitro artificial insemination method carry out, sperm and ovum can be obtained in vitro, Parent determine compared with
Easily, and there is a highly developed technology of hastening parturition, therefore obtain the retrospective family full-sibs of substantial amounts of Parent and be easier to realize.
However, Litopenaeus vannamei have embrace to knocking into the back, it is i.e. separate after mating, male shrimp spermatophore must be squeezed into female shrimp receptaculum seminis and could calculate real complete
Conclusion of the business is matched somebody with somebody, and many tail hero shrimps and female shrimp priority mating often occurs, and only 1 tail can squeeze into the phenomenon of receptaculum seminis, artificial orientation
Copulation occurs male shrimp not copulation again, or single tail copulation but spermatophore such as deflect at the unfavorable factor.In Litopenaeus vannamei reproductive process
Unfavorable factor greatly limit the preparation process of family full-sibs, hinder prawn molecular breeding and Parents selection-breeding work
Carry out.
The content of the invention:
The purpose of the present invention be for solve directional cross prepare family during deficiency, set up a kind of Litopenaeus vannamei with
Machine copulation parent's retroactive method.
Litopenaeus vannamei random mating parent's retroactive method of the present invention, it is characterised in that comprise the following steps:
It is prepared by a, male shrimp DNA library:Litopenaeus vannamei Broodstock after sexual maturity idiophase is separately raised, and male shrimp is in male shrimp ponds
Raise, female shrimp is raised in female shrimp ponds, and last is cut to swimmeret by all male shrimp in male shrimp ponds, then extract trip respectively
The DNA of swimming foot, the male shrimp DNA library of composition;
B, close shrimp random mating:In morning 7-11, select the full female shrimp for extending to afterbody of gonad and be put in male shrimp ponds,
Male shrimp and female shrimp pantogamy;
C, the spawning of female shrimp:In same day 18-21, female shrimp is pulled out, check whether receptaculum seminis squeezes into spermatophore, correctly beaten
Enter the female shrimp of spermatophore, each female shrimp for correctly squeezing into spermatophore lays eggs in being put into single water body;
D, take spermatophore:Check whether female shrimp lays eggs in independent water body, immediately by female shrimp receptaculum seminis after finding to lay eggs
Spermatophore is removed and cuts the swimmeret of the female shrimp that lays eggs;
E, parent review:The swimmeret DNA of the female shrimp of spawning is extracted, that is, obtains female parent gene group DNA;Remain in extracting spermatophore
Thick white spermatium DNA, using the stable prawn micro-satellite primers of rich polymorphism, amplified production enter performing PCR amplification,
And carry out matching identification with the result in male shrimp DNA library, the male shrimp DNA for matching is male parent gene group DNA, thereby determines that father
This.
The invention has the beneficial effects as follows:The present invention remains on numerous in female shrimp body with reference to spermatophore after Litopenaeus vannamei spawning
Feature is grown, at present more ripe prawn microsatellite paternity test method is added, Litopenaeus vannamei random mating, afterwards is realized
Carry out the purpose that parent reviews identification.Present invention utilizes the female shrimp success mating efficiency high of the production process of Litopenaeus vannamei is excellent
Point, overcomes the deficiency of orientation mating male and female not copulation or post-coitum spermatophore malposition, and the application of the inventive method will add
The preparation of fast Litopenaeus vannamei family full-sibs, contributes to promoting the carrying out of prawn molecular breeding work.
Specific embodiment:
Following examples are that the present invention is further illustrated, rather than limitation of the present invention.
Embodiment 1:
1st, prepared by male shrimp DNA library:Litopenaeus vannamei after sexual maturity idiophase is separately raised, and male shrimp is raised in male shrimp ponds
Support, female shrimp is raised in female shrimp ponds, by all 80 tail hero shrimps in male shrimp ponds, last is cut to swimmeret, in volume fraction
95% -20 DEG C of ethanol is preserved, and then extracts the DNA of swimmeret respectively, and individually (male 1,2,3 ... 80), the male shrimp DNA of composition for numbering
Storehouse;
2nd, close shrimp random mating:In morning 7-11, select full 15 tail of female shrimp for extending to afterbody of gonad and be put into male shrimp
Chi Zhong, male shrimp and female shrimp pantogamy;
3rd, female shrimp spawning:In same day 18-21,15 tails female shrimp is pulled out, check whether receptaculum seminis squeezes into spermatophore, obtain 10
Tail correctly squeezes into the female shrimp of spermatophore, and each female shrimp for correctly squeezing into spermatophore lays eggs in being put into single 1 barrel of water body;
4th, take spermatophore:After female shrimp drags for independent water body spawning, patrol once per 10-30 minutes, see whether female shrimp has produced
Spermatophore in female shrimp receptaculum seminis is removed after finding spawning and cuts the swimmeret of the female shrimp that lays eggs, female shrimp swimmeret, essence by ovum immediately
Pod is put in different test tubes, individually numbering (female 1,2,3 ... 10;Essence 1,2,3 ... 10), as far as possible collects thick white shape
Spermatium, Liquid nitrogen storage;
5th, parent reviews:The swimmeret DNA of the female shrimp of spawning is extracted, that is, obtains female parent gene group DNA (female 1,2,3 ...
10);The thick white spermatium remained in spermatophore is used to extract DNA, enters performing PCR using the prawn microsatellite sequence announced
Amplification, and carry out matching identification with the result in the male shrimp DNA library of step 1, concrete grammar authorizes patent of invention " to enter with reference to China
The multiple microsatellite identification system of row Litopenaeus vannamei Parentage determination and apply ZL201310340730.1 " described in method enter
OK, the male shrimp DNA for matching is male parent gene group DNA, 8 tail of successful match hero shrimp genomic DNA (numbering:Male 6,8,9,20,
45,54,78,79).Obtain the retrospective family full-sibs of 8 groups of parents, parent identifies success rate, and (10 tails are effectively laid eggs up to 80%
Female shrimp, 8 tail spermatophore DNA extraction successes).
Claims (1)
1. a kind of Litopenaeus vannamei random mating parent's retroactive method, it is characterised in that comprise the following steps:
It is prepared by a, male shrimp DNA library:Litopenaeus vannamei Broodstock after sexual maturity idiophase is separately raised, and male shrimp is raised in male shrimp ponds
Support, female shrimp is raised in female shrimp ponds, and last is cut to swimmeret by all male shrimp in male shrimp ponds, then extract swimming respectively
The DNA of foot, the male shrimp DNA library of composition;
B, close shrimp random mating:In morning 7-11, select the full female shrimp for extending to afterbody of gonad and be put in male shrimp ponds, male shrimp
With female shrimp pantogamy;
C, the spawning of female shrimp:In same day 18-21, female shrimp is pulled out, check whether receptaculum seminis squeezes into spermatophore, correctly squeezed into essence
The female shrimp of pod, each female shrimp for correctly squeezing into spermatophore lay eggs in being put into single water body;
D, take spermatophore:Check whether female shrimp lays eggs in independent water body, immediately by the spermatophore in female shrimp receptaculum seminis after finding to lay eggs
Remove and cut the swimmeret of female shrimp of laying eggs;
E, parent review:The swimmeret DNA of the female shrimp of spawning is extracted, that is, obtains female parent gene group DNA;What is remained in extracting spermatophore is white
The DNA of the sticky spermatium of color, is entered performing PCR amplification using Litopenaeus vannamei micro-satellite primers, and is entered with the result in male shrimp DNA library
Row matching identification, the male shrimp DNA for matching are male parent gene group DNA, thereby determine that male parent.
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CN201611201011.1A CN106577425B (en) | 2016-12-22 | 2016-12-22 | A kind of litopenaeus vannamei panmixia parent's retroactive method |
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Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107581123A (en) * | 2017-09-19 | 2018-01-16 | 中国科学院南海海洋研究所 | A kind of Environment of Litopenaeus vannamei Low semi-directional mating family full-sibs structure and parent determine method |
CN113100134A (en) * | 2021-04-13 | 2021-07-13 | 广东金阳生物技术有限公司 | Whole sibling family construction and parent determination method for Litopenaeus vannamei male parent semi-directional mating |
Families Citing this family (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN111110718B (en) * | 2020-01-03 | 2021-08-27 | 中国科学院南海海洋研究所 | Special pesticide for prawn enterocytocele and preparation method and application thereof |
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CN102399776A (en) * | 2011-09-02 | 2012-04-04 | 中国水产科学研究院黄海水产研究所 | Microsatellite marker quadruple PCR primer for identifying Penaeus chinesis family and identification method |
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Cited By (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN107581123A (en) * | 2017-09-19 | 2018-01-16 | 中国科学院南海海洋研究所 | A kind of Environment of Litopenaeus vannamei Low semi-directional mating family full-sibs structure and parent determine method |
CN107581123B (en) * | 2017-09-19 | 2019-10-18 | 中国科学院南海海洋研究所 | A kind of building of litopenaeus vannamei semi-directional mating family full-sibs and parent determine method |
CN113100134A (en) * | 2021-04-13 | 2021-07-13 | 广东金阳生物技术有限公司 | Whole sibling family construction and parent determination method for Litopenaeus vannamei male parent semi-directional mating |
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