CN106106279A - A kind of raising bream, Carnis Megalobramae triploid incubation rate method - Google Patents
A kind of raising bream, Carnis Megalobramae triploid incubation rate method Download PDFInfo
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- CN106106279A CN106106279A CN201610496433.XA CN201610496433A CN106106279A CN 106106279 A CN106106279 A CN 106106279A CN 201610496433 A CN201610496433 A CN 201610496433A CN 106106279 A CN106106279 A CN 106106279A
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- 238000000034 method Methods 0.000 title claims abstract description 43
- 208000026487 Triploidy Diseases 0.000 title claims abstract description 28
- 241001519451 Abramis brama Species 0.000 title claims abstract description 23
- 238000011534 incubation Methods 0.000 title claims abstract description 13
- NKANXQFJJICGDU-QPLCGJKRSA-N Tamoxifen Chemical compound C=1C=CC=CC=1C(/CC)=C(C=1C=CC(OCCN(C)C)=CC=1)/C1=CC=CC=C1 NKANXQFJJICGDU-QPLCGJKRSA-N 0.000 claims abstract description 56
- 239000000243 solution Substances 0.000 claims abstract description 34
- 238000002347 injection Methods 0.000 claims abstract description 29
- 239000007924 injection Substances 0.000 claims abstract description 29
- 229960001603 tamoxifen Drugs 0.000 claims abstract description 28
- 239000003112 inhibitor Substances 0.000 claims abstract description 23
- 229940011871 estrogen Drugs 0.000 claims abstract description 18
- 239000000262 estrogen Substances 0.000 claims abstract description 18
- 230000002706 hydrostatic effect Effects 0.000 claims abstract description 16
- 208000035199 Tetraploidy Diseases 0.000 claims abstract description 15
- 238000003825 pressing Methods 0.000 claims abstract description 13
- 230000035939 shock Effects 0.000 claims abstract description 13
- 238000002360 preparation method Methods 0.000 claims abstract description 11
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 16
- 210000004602 germ cell Anatomy 0.000 claims description 14
- MUMGGOZAMZWBJJ-DYKIIFRCSA-N Testostosterone Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 MUMGGOZAMZWBJJ-DYKIIFRCSA-N 0.000 claims description 12
- 239000010413 mother solution Substances 0.000 claims description 12
- 230000008774 maternal effect Effects 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- 239000003205 fragrance Substances 0.000 claims description 6
- 229960003604 testosterone Drugs 0.000 claims description 6
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 0.000 claims description 4
- 108010062540 Chorionic Gonadotropin Proteins 0.000 claims description 4
- 102000011022 Chorionic Gonadotropin Human genes 0.000 claims description 4
- 235000007164 Oryza sativa Nutrition 0.000 claims description 4
- 229910052801 chlorine Inorganic materials 0.000 claims description 4
- 239000000460 chlorine Substances 0.000 claims description 4
- 229940015047 chorionic gonadotropin Drugs 0.000 claims description 4
- GKIRPKYJQBWNGO-OCEACIFDSA-N clomifene Chemical compound C1=CC(OCCN(CC)CC)=CC=C1C(\C=1C=CC=CC=1)=C(\Cl)C1=CC=CC=C1 GKIRPKYJQBWNGO-OCEACIFDSA-N 0.000 claims description 4
- 229960003608 clomifene Drugs 0.000 claims description 4
- 229940088597 hormone Drugs 0.000 claims description 4
- 239000005556 hormone Substances 0.000 claims description 4
- 230000008775 paternal effect Effects 0.000 claims description 4
- 230000000149 penetrating effect Effects 0.000 claims description 4
- 230000002085 persistent effect Effects 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 235000009566 rice Nutrition 0.000 claims description 4
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 239000012895 dilution Substances 0.000 claims description 2
- 238000010790 dilution Methods 0.000 claims description 2
- 238000004821 distillation Methods 0.000 claims description 2
- 230000000762 glandular Effects 0.000 claims description 2
- 150000002576 ketones Chemical class 0.000 claims description 2
- 230000032696 parturition Effects 0.000 claims description 2
- 210000001550 testis Anatomy 0.000 claims description 2
- 240000007594 Oryza sativa Species 0.000 claims 1
- 238000009395 breeding Methods 0.000 abstract description 3
- 238000009396 hybridization Methods 0.000 abstract description 3
- 230000001488 breeding effect Effects 0.000 abstract description 2
- 239000008239 natural water Substances 0.000 abstract description 2
- 230000003247 decreasing effect Effects 0.000 abstract 1
- 241000251468 Actinopterygii Species 0.000 description 10
- 241000209094 Oryza Species 0.000 description 3
- 210000000349 chromosome Anatomy 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 235000013372 meat Nutrition 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 244000025254 Cannabis sativa Species 0.000 description 1
- 230000004543 DNA replication Effects 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 241001275890 Megalobrama amblycephala Species 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000013505 freshwater Substances 0.000 description 1
- 230000009931 harmful effect Effects 0.000 description 1
- 238000002386 leaching Methods 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- 230000011278 mitosis Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
Abstract
The invention discloses and a kind of improve bream, the method for Carnis Megalobramae triploid incubation rate, comprise the steps: the outfit of tamoxifen inhibitor, the preparation of tamoxifen injection, injection treatment female parent, cold shock+hydrostatic pressing process, the preparation of estrogen inhibitors solution, obtain bream, Carnis Megalobramae male tetraploid pure lines, use the mode backcrossed to obtain triploid.By the method, bream, the Carnis Megalobramae triploid pure lines of 100% can be obtained, mutually for hybridization frequently, the triploid pure lines that purity is higher, offspring is more stable can be obtained, compared to existing technology, decreasing the trouble of breeding and seed selection, more importantly matter all to Natural Water product and ecological environment are pollution-free.
Description
Technical field
The present invention relates to the triploid mating system of bream, Carnis Megalobramae, be specifically related to a kind of raising bream, Carnis Megalobramae triploid incubation rate
Method.
Background technology
Bream cries again Changchun bream, grass bream etc., and triangular bream is divided into again triangular bream and Megalobrama amblycephala, and its growth is rapider, adaptable,
Feeding habits are wide, and meat is soft, delicious flavour, are one of main cultured freshwater fishes of China.Triploid bream, the sterile feature of Carnis Megalobramae,
It is made to show significantly more advantage at aspects such as the speed of growth, resistance, disease resistance and meats.In prior art, no
Educating triploid fish is to be obtained with liploid fish interspecies crossing by tetraploid fish, but to obtain the triploid fish of more than 98%, then
It is accomplished by setting up the tetraploid fish system of inheritance stability, very high purity.
In genetics-breeding in fish, cold heat is generally utilized to suffer a shock, or the first time by method suppression germ cell such as hybridization
Mitosis or directly chromosome nonhomologous double, and prepare homology or allotrtraploid fish, and wherein cold shock is usually and hydrostatic
Pressure process together, it is difficult to obtain higher degree tetraploid system, although and the mode of heat shock is easily operated, equally, it is difficult to obtain
Obtain the tetraploid system that purity is high；Although and the method purity height hybridized, but success rate is low, the cycle is long.Additionally, these methods above-mentioned
The tetraploid fish of preparation, some can be educated, some can not be educated again, also likely to be present gene simultaneously and mixes phenomenon (allotetraploid),
Easily cause bio-hazard.
Summary of the invention
The deficiency existed for above-mentioned prior art, it is an object of the invention to provide the stable raising bream of a kind of method, triangular bream
The method of fish triploid incubation rate.
For reaching above-mentioned purpose, technical scheme is as follows:
A kind of improve bream, the method for Carnis Megalobramae triploid incubation rate, comprise the steps:
(1) tamoxifen inhibitor is equipped with: take the tamoxifen powder that purity is more than 95%, and the ethanol solution of addition 90% is carried out
Dissolving, make mother solution, wherein tamoxifen content is about 50～100mg/ml, stand-by；
(2) preparation of tamoxifen injection: take above-mentioned mother solution and be added directly in the odinagogue solution prepared, its ratio
Join for 1ml tamoxifen mother solution in the odinagogue solution prepared of 250～500ml, make injection stand-by；Hasten parturition
Agent solution is formulated with 0.9% normal saline dilution by chorionic-gonadotropin hormone, odinagogue solution medium staple chorionic gonadotropin
The dosage of glandular hormone is 5-10 IU/g；
(3) injection treatment is maternal: before maternal artificial spawning, routinely induced spawning method female parent is carried out manual injection he not
Former times sweet smell injection, wherein the consumption of tamoxifen injection is about 40～160 μ g/kg；
(4) cold shock+hydrostatic pressing processes: when a spilting of an egg, uses the method that cold shock combines with hydrostatic pressing, through note
The female parent penetrating process puts into, with the germ cell obtained without the paternal hybrid of any process, the cold water that temperature is-1～1.0 DEG C
In, hydrostatic pressing pressure is 250～340Kg/cm2, the persistent period is 3～5min；
(5) preparation of estrogen inhibitors solution: clomifene is dissolved in the ethanol of 80%～90% together with testosterone, then adds distillation
Water is configured to estrogen inhibitors solution, and the concentration that wherein estrogen inhibitors Chlorine in Solution rice is fragrant is 150～250mg/l, testis
The concentration of ketone is 40～120mg/l；
(6) taking the germ cell after step (4) processes and put in the estrogen inhibitors solution as prepared by step (5), room temperature soaks
Bubble 10～30s, then takes out and is put in normal incubation in couveuse；Thus obtain 100% bream, Carnis Megalobramae male tetraploid pure lines；
(7) triploid acquisition: take the male tetraploid of step (6) as male parent, takes common diploid as female parent, uses back
The mode handed over, it is thus achieved that 100% triploid.
Advantages of the present invention: 1, inject tamoxifen in the present invention in female parent, play suppression DNA replication dna and promote male
While dual function, the germ cell First cleavage time is put off 3-5min, and improves all germ cell First cleavages
Synchronization degree (up to 99%), ensure and established the basis of chromosome doubling for operating next step offer time；2, in germ cell
Spilting of an egg uses cold shock and relatively low hydrostatic pressing (less than conventional 350kg/cm2), and the shorter process time (be less than
5min), not only effectively reduce the harmful effect to germ cell, improve the survival rate of germ cell；3, estrogen inhibitors solution leaching
Bubble, strengthens again and ensures male conversion ratio；4, backcross with original diploid female, effectively with the male tetraploid of pure lines
Ensure that triploid purity.Thereby through the method, bream, the Carnis Megalobramae triploid pure lines of 100% can be obtained, mutually hybridization frequently and
Speech, can obtain the triploid pure lines that purity is higher, offspring is more stable, compared to existing technology, decrease the fiber crops of breeding and seed selection
Tired, more importantly matter all to Natural Water product and ecological environment are pollution-free.
Detailed description of the invention
In conjunction with specific embodiment, the invention will be further described.
Embodiment one
The method improving bream triploid incubation rate, comprises the steps:
(1) tamoxifen inhibitor is equipped with: take the tamoxifen powder that purity is more than 95%, and the ethanol solution of addition 90% is carried out
Dissolving, make mother solution, wherein tamoxifen content is about 100mg/ml, stand-by；
(2) preparation of tamoxifen injection: take above-mentioned mother solution and be added directly in the odinagogue solution prepared, its ratio
Joining for 1ml tamoxifen mother solution in the odinagogue solution prepared of 250ml, make injection, in injection, he is not
The former times concentration of sweet smell is about 0.4mg/ml, stand-by；
(3) injection treatment is maternal: before maternal artificial spawning, routinely induced spawning method female parent is carried out manual injection he not
Former times sweet smell injection, wherein the consumption of tamoxifen injection is about 120 μ g/kg；
(4) cold shock+hydrostatic pressing processes: when a spilting of an egg, uses the method that cold shock combines with hydrostatic pressing, through note
The female parent penetrating process is put in the cold water that temperature is 0 DEG C with the germ cell obtained without the paternal hybrid of any process, hydrostatic
Pressure pressure is 310Kg/cm2, the persistent period is 3min；
(5) preparation of estrogen inhibitors solution: clomifene is dissolved in the ethanol of 90% together with testosterone, then add distilled water and join
Making estrogen inhibitors solution, the concentration that wherein estrogen inhibitors Chlorine in Solution rice is fragrant is 150mg/l, and the concentration of testosterone is
40mg/l；
(6) taking the germ cell after step (4) processes and put in the estrogen inhibitors solution as prepared by step (5), room temperature soaks
Bubble 30s, then takes out and is put in normal incubation in couveuse；Thus obtain bream male tetraploid pure lines；
(7) triploid acquisition: take the male tetraploid of step (6) as male parent, takes common diploid as female parent, uses back
The mode handed over, it is thus achieved that 100% triploid.
Embodiment two
The method improving Carnis Megalobramae triploid incubation rate, comprises the steps:
(1) tamoxifen inhibitor is equipped with: take the tamoxifen powder that purity is more than 95%, and the ethanol solution of addition 90% is carried out
Dissolving, make mother solution, wherein tamoxifen content is about 100mg/ml, stand-by；
(2) preparation of tamoxifen injection: take above-mentioned mother solution and be added directly in the odinagogue solution prepared, its ratio
Joining for 1ml tamoxifen mother solution in the odinagogue solution prepared of 500ml, make injection, in injection, he is not
The former times concentration of sweet smell is 0.2mg/ml, stand-by；
(3) injection treatment is maternal: before maternal artificial spawning, routinely induced spawning method female parent is carried out manual injection he not
Former times sweet smell injection, wherein the consumption of tamoxifen injection is about 160 μ g/kg；
(4) cold shock+hydrostatic pressing processes: when a spilting of an egg, uses the method that cold shock combines with hydrostatic pressing, through note
The female parent penetrating process is put in the cold water that temperature is-1 DEG C with the germ cell obtained without the paternal hybrid of any process, hydrostatic
Pressure pressure is 280Kg/cm2, the persistent period is 3min；
(5) preparation of estrogen inhibitors solution: clomifene is dissolved in the ethanol of 90% together with testosterone, then add distilled water and join
Making estrogen inhibitors solution, the concentration that wherein estrogen inhibitors Chlorine in Solution rice is fragrant is 250mg/l, and the concentration of testosterone is
120mg/l；
(6) taking the germ cell after step (4) processes and put in the estrogen inhibitors solution as prepared by step (5), room temperature soaks
Bubble 10s, then takes out and is put in normal incubation in couveuse；Thus obtain Carnis Megalobramae male tetraploid pure lines；
(7) triploid acquisition: take the male tetraploid of step (6) as male parent, takes common diploid as female parent, uses back
The mode handed over, it is thus achieved that 100% triploid.
Above-mentioned two embodiment is only the two of the preferred version of the present invention, can not limit the scope of the invention, and appoints
Meaning is equal to the present invention or similar technological means is considered as protection scope of the present invention.Hereinafter enclose part processing mode of the present invention
And result statistics.
Attached: bream, Carnis Megalobramae triploid are bred as result statistics
Note: the above results is soak at room temperature 10S under conditions of the concentration of tamoxifen is 0.2mg/ml in injection, hydrostatic pressing
Process 3 minutes acquired results.
Claims (1)
1. improve bream, a method for Carnis Megalobramae triploid incubation rate, comprise the steps:
(1) tamoxifen inhibitor is equipped with: take the tamoxifen powder that purity is more than 95%, and the ethanol solution of addition 90% is carried out
Dissolving, make mother solution, wherein tamoxifen content is about 50～100mg/ml, stand-by；
(2) preparation of tamoxifen injection: take above-mentioned mother solution and be added directly in the odinagogue solution prepared, its ratio
Join for 1ml tamoxifen mother solution in the odinagogue solution prepared of 250～500ml, make injection stand-by；Hasten parturition
Agent solution is formulated with 0.9% normal saline dilution by chorionic-gonadotropin hormone, odinagogue solution medium staple chorionic gonadotropin
The dosage of glandular hormone is 5-10 IU/g；
(3) injection treatment is maternal: before maternal artificial spawning, routinely induced spawning method female parent is carried out manual injection he not
Former times sweet smell injection, wherein the consumption of tamoxifen injection is about 40～160 μ g/kg；
(4) cold shock+hydrostatic pressing processes: when a spilting of an egg, uses the method that cold shock combines with hydrostatic pressing, through note
The female parent penetrating process puts into, with the germ cell obtained without the paternal hybrid of any process, the cold water that temperature is-1～1.0 DEG C
In, hydrostatic pressing pressure is 250～340Kg/cm2, the persistent period is 3～5min；
(5) preparation of estrogen inhibitors solution: clomifene is dissolved in the ethanol of 80%～90% together with testosterone, then adds distillation
Water is configured to estrogen inhibitors solution, and the concentration that wherein estrogen inhibitors Chlorine in Solution rice is fragrant is 150～250mg/l, testis
The concentration of ketone is 40～120mg/l；
(6) taking the germ cell after step (4) processes and put in the estrogen inhibitors solution as prepared by step (5), room temperature soaks
Bubble 10～30s, then takes out and is put in normal incubation in couveuse；Thus obtain bream, Carnis Megalobramae male tetraploid pure lines；
(7) triploid acquisition: take the male tetraploid of step (6) as male parent, takes common diploid as female parent, uses back
The mode handed over, it is thus achieved that 100% triploid.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610496433.XA CN106106279A (en) | 2016-06-30 | 2016-06-30 | A kind of raising bream, Carnis Megalobramae triploid incubation rate method |
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| CN201610496433.XA CN106106279A (en) | 2016-06-30 | 2016-06-30 | A kind of raising bream, Carnis Megalobramae triploid incubation rate method |
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| CN106106279A true CN106106279A (en) | 2016-11-16 |
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| CN201610496433.XA Pending CN106106279A (en) | 2016-06-30 | 2016-06-30 | A kind of raising bream, Carnis Megalobramae triploid incubation rate method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107711615A (en) * | 2017-10-31 | 2018-02-23 | 金华职业技术学院 | A kind of method that sterile fish is obtained using immersion fish-egg |
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- 2016-06-30 CN CN201610496433.XA patent/CN106106279A/en active Pending
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107711615A (en) * | 2017-10-31 | 2018-02-23 | 金华职业技术学院 | A kind of method that sterile fish is obtained using immersion fish-egg |
| CN107711615B (en) * | 2017-10-31 | 2020-08-11 | 金华职业技术学院 | Method for obtaining sterile fish by soaking fish eggs |
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| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20161116 |