CN1951183A - Daiquzu pseudosciaena crocea and minyuedongzu cultured pseudosciaena crocea cross breeding method - Google Patents
Daiquzu pseudosciaena crocea and minyuedongzu cultured pseudosciaena crocea cross breeding method Download PDFInfo
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- CN1951183A CN1951183A CNA2006101352605A CN200610135260A CN1951183A CN 1951183 A CN1951183 A CN 1951183A CN A2006101352605 A CNA2006101352605 A CN A2006101352605A CN 200610135260 A CN200610135260 A CN 200610135260A CN 1951183 A CN1951183 A CN 1951183A
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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
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- 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 relates to a cross method between large yellow Crocker and GuangDong large yellow Crocker. Wherein, their crossed generation has high grow speed and the application in long transmission; the invention selects GuangDong large yellow crocker as father, whose belly can be pressed to flow sperm; preparing dilute solution; compressing its belly to absorb sperm, whose activity is higher than 90% to be frozen and stored; mixing the dilute solution and anti-freeze agent, adding sperm, storing in frozen storage tube, sealing and putting in 4Deg. C; arranging it above the liquid nitrogen, reducing temperature and moving it into liquid nitrogen; selecteing the wide GuangDong large yellow crocker as mother; when its ovogen is mature, defreezing the sperm, and taking the freeze storage tube from liquid nitrogen to be put into room-temperature fresh water to be defrozen, activating it with seawater; artificial inseminating and washing zygote with seawater, removing sink zygote, and charging air and cultivating.
Description
Technical field
The present invention relates to a kind of large yellow Crocker, especially relate to the cross method of a kind of tai-chu race large yellow Crocker and east Guangdong cultured large yellow croaker.
Background technology
There are 3 geographical population in the coastal large yellow Crocker of China, and there is notable difference in they at aspects such as life-span, age at sexual maturity and population age compositions.With Northern Part of Zhejiang Mount Tai Qu Yang is that the tai-chu race large yellow Crocker of representative has longest-lived (raun the highest 29 ages, 27 ages of milter), (milter was 3 ages to a large amount of age at sexual maturity late, 3 ages of raun and 4 ages), population forms the most complicatedly, the residue population is accounting for characteristics such as stable advantage; With western island in Guangdong Province, Guangdong is that the island in Guangdong Province family large yellow Crocker of representative has the shortest (milter the highest 8 ages in life-span, 9 ages of raun), a large amount of age at sexual maturity early (just has the individual beginning of minority sexual maturity 1 age, a large amount of individual in 2-3 rheological properties maturation), population is formed simple and the recruit population proportion increases; With the characteristics of the northern official Jing Yang in the Fujian east Guangdong large yellow Crocker that is representative then between tai-chu race and island in Guangdong Province family.Significant group difference is indicating that carrying out large yellow Crocker intraspecific hybridization is a very significant job, and the hereditary difference between the parent is remarkable more, and the hybrid vigour of filial generation also may be got over economically valuable and using value.
The not only residing geographical position of large yellow Crocker difference, the sexually matured time is also asynchronous.Because the long-distance transport of large yellow Crocker adult fish is very difficult, these all seriously hinder carrying out smoothly of large yellow Crocker population intermolecular hybrid.
The applicant provides a kind of Shishou fish sperm to excite the diplontic abductive approach of large yellow croaker gynogenesis in the application for a patent for invention of publication number for CN1586430, this method adopts artificial insemination, and the sperm of equal not male parent of the same race in the hybrid combination is mixed with the ovum of large yellow Crocker; After smart ovum mixing, with sea-water activated: fertilized egg is put into pressure vessel, exert pressure; Stop to exert pressure when reaching target pressurizing force 38～55MPa; Inducing of pressure shock finished in step-down behind lasting 1～30min.
Summary of the invention
The objective of the invention is at tai-chu race and 2 population large yellow Crocker of east Guangdong cultured large yellow croaker in gonad maturity nonsynchronous problem on the time, provide that a kind of filial generation has that growth is fast, strong stress resistance, be suitable for the tai-chu race large yellow Crocker of advantages such as transporting for long-distance and the cross method of east Guangdong cultured large yellow croaker.
Concrete operations of the present invention are as follows:
1) chooses east Guangdong cultured large yellow croaker milter male parent, gently press belly to have milky seminal fluid outflow person to continue to employ;
2) configuration spermatozoa diluent;
3) push the milter belly, draw the milky seminal fluid of no blood anuria, be used for freezing preservation 90% above person through the microscopy sperm viability;
4) spermatozoa diluent and antifreeze mixing are added sperm again, be stored in behind the mixing in the frozen pipe, divide the frozen seal of tube that installs to be placed in 4 ℃ of environment, place cooling back, liquid nitrogen top to move into liquid nitrogen again and preserve standby;
5) choose the wild large yellow Crocker raun of tai-chu race female parent, hasten parturition;
6) begin the seminal fluid that thaws when large yellow Crocker female parent maturation of ovum, drop in the room-temperature fresh water immediately after the frozen pipe that sperm is housed is taken out from liquid nitrogen and thaw, the sperm after thawing carries out artificial insemination simultaneously with sea-water activated;
7) after the artificial insemination, wash fertilized egg repeatedly, in time remove the bad ovum of sinking, inflation hatching with fresh seawater.
In step 1), it is complete, healthy that described east Guangdong cultured large yellow croaker milter male parent is preferably the bodily form, the east Guangdong cultured large yellow croaker milter male parent of belly softness.
In step 2) described in the configuration spermatozoa diluent contain glucose 1.45g with every 100mL spermatozoa diluent; Sodium citrate 0.2g; KCl0.03g; BSA (bSA) 3g; Regulating pH is 5.6～6.0.
In step 3), sperm: spermatozoa diluent: antifreeze=3: 6: 1, wherein antifreeze uses methyl-sulfoxide (DMSO).
In step 4), in proportion dilution and antifreeze mixing are added sperm again, be stored in behind the mixing in the frozen pipe of 1.8mL, behind the frozen seal of tube that branch installs, place 4 ℃ of environment cooling balance 40min, place again to move into liquid nitrogen behind the liquid nitrogen top cooling balance 30min and preserve standby.
In step 5), the wild large yellow Crocker raun of described tai-chu race female parent choose the bodily form complete, move about active, the belly softness, the wild large yellow Crocker raun of the little red tai-chu race of cloacal aperture female parent is heavily injected 1～3ug LHRH-A by every Kg fish
2Hasten parturition, 22～24 ℃ of water temperatures, the hormone effect phase is 36h.
In step 7), preferably behind artificial insemination 5～10min, wash fertilized egg repeatedly with fresh seawater.
The present invention at first analyzes the genetic background of 4 colonies of large yellow Crocker, detection and affirmation by modern molecular biology techniques such as RAPD, ISSR, find with the wild large yellow Crocker of tai-chu race to be female parent by further experiment, the east Guangdong cultured large yellow croaker is that offspring's hybrid vigour that hybrid combination produced of male parent is the most remarkable, does not exist reproduction to isolate.Determined that with the wild large yellow Crocker of tai-chu race be the hybrid combination that maternal east Guangdong cultured large yellow croaker is a male parent.
The method of using the sperm freezing preservation not only can overcome individual sexually matured time difference between different population, also can overcome the residing spatial diversity of different population, thereby can effectively solve the difficulty of the isolation of large yellow Crocker intraspecific hybridization on space-time.
Advantage of the present invention can reduce:
Hybrid combination be with the early stage experimental result be foundation, guarantee that the hybrid vigour of hybrid combination is the most remarkable;
2. the method for using the sperm freezing preservation has overcome the wild large yellow Crocker of tai-chu race and east Guangdong cultured large yellow croaker difference on the sexual maturity time, has solved both isolation geographically simultaneously;
3. the offspring that produces of hybridization has that growth is fast, strong stress resistance, is suitable for advantage such as long-distance transport.
Embodiment
1. choose east Guangdong cultured large yellow croaker milter 2 tails that the individual bodily form is complete, healthy, belly is soft in Fujian, gently press belly to have milky seminal fluid outflow person to continue to employ.
2. configuration spermatozoa diluent.Every 100mL spermatozoa diluent contains: glucose 1.45g; Sodium citrate 0.2g; KCL0.03g; BSA (bSA) 3g; Regulate pH to 5.6～6.0.
3. push the milter belly, draw the milky seminal fluid of no blood anuria, 91%, continue to employ through the microscopy sperm viability.Sperm: dilution: the ratio of antifreeze is 3: 6: 1, and wherein antifreeze uses methyl-sulfoxide (DMSO).
4. in proportion dilution and antifreeze mixing are added sperm again, thoroughly be stored in the frozen pipe of 1.8mL behind the mixing.After dividing the frozen seal of tube install, place 4 ℃ of environment cooling balance 40min, place again to move into liquid nitrogen behind the liquid nitrogen top cooling balance 30min and preserve.The seminal fluid of freezing preservation is transported to the producing region of the wild large yellow Crocker of tai-chu race---Zhejiang.
4. choose the bodily form complete, move about active, belly softness, wild large yellow Crocker raun 5 tails of the little red tai-chu race of cloacal aperture.Heavily inject 2ug LHRH-A by every Kg fish
2Hasten parturition.23 ℃ of water temperatures, the hormone effect phase is about 36h.
5. when large yellow Crocker female parent maturation of ovum, begin the seminal fluid that thaws, drop in the room-temperature fresh water immediately after the frozen pipe that sperm is housed is taken out from liquid nitrogen and thaw.Sperm after thawing is carried out at artificial insemination simultaneously with sea-water activated.
6. behind the artificial insemination 50min, wash fertilized egg repeatedly, in time remove the bad ovum of sinking, inflation hatching with fresh seawater.
Claims (8)
1. the cross method of tai-chu race large yellow Crocker and east Guangdong cultured large yellow croaker is characterized in that its operation is as follows:
1) chooses east Guangdong cultured large yellow croaker milter male parent, gently press belly to have milky seminal fluid outflow person to continue to employ;
2) configuration spermatozoa diluent;
3) extruding milter belly is drawn the milky seminal fluid, is used for freezing preservation through the microscopy sperm viability 90% above person;
4) spermatozoa diluent and antifreeze mixing are added sperm again, be stored in behind the mixing in the frozen pipe, divide the frozen seal of tube that installs to be placed in 4 ℃ of environment, place cooling back, liquid nitrogen top to move into liquid nitrogen again and preserve standby;
5) choose the wild large yellow Crocker raun of tai-chu race female parent, hasten parturition;
6) begin the seminal fluid that thaws when large yellow Crocker female parent maturation of ovum, drop in the room-temperature fresh water immediately after the frozen pipe that sperm is housed is taken out from liquid nitrogen and thaw, the sperm after thawing carries out artificial insemination simultaneously with sea-water activated;
7) after the artificial insemination, wash fertilized egg repeatedly, in time remove the bad ovum of sinking, inflation hatching with fresh seawater.
2. the cross method of tai-chu race large yellow Crocker as claimed in claim 1 and east Guangdong cultured large yellow croaker, it is characterized in that in step 1), described east Guangdong cultured large yellow croaker milter male parent is that the bodily form is complete, and is healthy, the east Guangdong cultured large yellow croaker milter male parent of belly softness.
3. the cross method of tai-chu race large yellow Crocker as claimed in claim 1 and east Guangdong cultured large yellow croaker is characterized in that in step 2) described in the configuration spermatozoa diluent contain glucose 1.45g with every 100mL spermatozoa diluent; Sodium citrate 0.2g; KCl0.03g; BSA 3g; Regulating pH is 5.6～6.0.
4. the cross method of tai-chu race large yellow Crocker as claimed in claim 1 and east Guangdong cultured large yellow croaker is characterized in that in step 3) sperm: spermatozoa diluent: antifreeze=3: 6: 1.
5. as the cross method of claim 1 or 4 described tai-chu race large yellow Crocker and east Guangdong cultured large yellow croaker, it is characterized in that antifreeze is a methyl-sulfoxide.
6. the cross method of tai-chu race large yellow Crocker as claimed in claim 1 and east Guangdong cultured large yellow croaker, it is characterized in that in step 4), in proportion dilution and antifreeze mixing are added sperm again, be stored in behind the mixing in the frozen pipe of 1.8mL, after dividing the frozen seal of tube that installs, place 4 ℃ of environment cooling balance 40min, place again to move into liquid nitrogen behind the liquid nitrogen top cooling balance 30min and preserve standby.
7. the cross method of tai-chu race large yellow Crocker as claimed in claim 1 and east Guangdong cultured large yellow croaker, it is characterized in that in step 5), the wild large yellow Crocker raun of described tai-chu race female parent choose the bodily form complete, move about active, the belly softness, the wild large yellow Crocker raun of the little red tai-chu race of cloacal aperture female parent is heavily injected 1～3ug LHRH-A by every Kg fish
2Hasten parturition, 22～24 ℃ of water temperatures, the hormone effect phase is 36h.
8. the cross method of tai-chu race large yellow Crocker as claimed in claim 1 and east Guangdong cultured large yellow croaker is characterized in that in step 7), preferably behind artificial insemination 5～10min, washes fertilized egg repeatedly with fresh seawater.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2006101352605A CN100484397C (en) | 2006-11-28 | 2006-11-28 | Daiquzu pseudosciaena crocea and minyuedongzu cultured pseudosciaena crocea cross breeding method |
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| CNB2006101352605A CN100484397C (en) | 2006-11-28 | 2006-11-28 | Daiquzu pseudosciaena crocea and minyuedongzu cultured pseudosciaena crocea cross breeding method |
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| CN1951183A true CN1951183A (en) | 2007-04-25 |
| CN100484397C CN100484397C (en) | 2009-05-06 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101965828A (en) * | 2010-08-31 | 2011-02-09 | 广西壮族自治区水产研究所 | Method for cryopreserving lutjanus argentimaculatus sperm |
| CN101502256B (en) * | 2009-03-11 | 2012-02-15 | 宁波大学 | Ultra low temperature cryopreservation method for sperm of large yellow crocker and cryopreservation device |
| CN106172117A (en) * | 2016-07-14 | 2016-12-07 | 浙江省海洋水产研究所 | Carnis Pseudosciaenae raun and Carnis Pseudosciaenae milter cross breeding method |
| CN111869656A (en) * | 2020-07-29 | 2020-11-03 | 中国水产科学研究院黄海水产研究所 | Ultralow-temperature cryopreservation method for thamnaconus modestus sperms |
| CN112535139A (en) * | 2020-11-18 | 2021-03-23 | 浙江省海洋水产研究所 | Sebastes bastes birth seed seedling device |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1295956C (en) * | 2004-07-27 | 2007-01-24 | 中国水产科学研究院黄海水产研究所 | Practicalization method for frozen preserving sperm of fish |
-
2006
- 2006-11-28 CN CNB2006101352605A patent/CN100484397C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101502256B (en) * | 2009-03-11 | 2012-02-15 | 宁波大学 | Ultra low temperature cryopreservation method for sperm of large yellow crocker and cryopreservation device |
| CN101965828A (en) * | 2010-08-31 | 2011-02-09 | 广西壮族自治区水产研究所 | Method for cryopreserving lutjanus argentimaculatus sperm |
| CN101965828B (en) * | 2010-08-31 | 2013-09-04 | 广西壮族自治区水产研究所 | Method for cryopreserving lutjanus argentimaculatus sperm |
| CN106172117A (en) * | 2016-07-14 | 2016-12-07 | 浙江省海洋水产研究所 | Carnis Pseudosciaenae raun and Carnis Pseudosciaenae milter cross breeding method |
| CN106172117B (en) * | 2016-07-14 | 2019-08-06 | 浙江省海洋水产研究所 | Little yellow croaker raun and Larimichthys crocea milter cross breeding method |
| CN111869656A (en) * | 2020-07-29 | 2020-11-03 | 中国水产科学研究院黄海水产研究所 | Ultralow-temperature cryopreservation method for thamnaconus modestus sperms |
| CN112535139A (en) * | 2020-11-18 | 2021-03-23 | 浙江省海洋水产研究所 | Sebastes bastes birth seed seedling device |
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| CN100484397C (en) | 2009-05-06 |
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