CN105941332A - Method for increasing breeding rate of Channa argus triploid - Google Patents
Method for increasing breeding rate of Channa argus triploid Download PDFInfo
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- CN105941332A CN105941332A CN201610497626.7A CN201610497626A CN105941332A CN 105941332 A CN105941332 A CN 105941332A CN 201610497626 A CN201610497626 A CN 201610497626A CN 105941332 A CN105941332 A CN 105941332A
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- triploid
- ophicephalus argus
- female parent
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- 241001597062 Channa argus Species 0.000 title claims abstract description 47
- 238000000034 method Methods 0.000 title claims abstract description 36
- 208000026487 Triploidy Diseases 0.000 title claims abstract description 27
- 238000009395 breeding Methods 0.000 title claims abstract description 10
- 230000001488 breeding effect Effects 0.000 title claims abstract description 9
- 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 38
- 229940011871 estrogen Drugs 0.000 claims abstract description 19
- 239000000262 estrogen Substances 0.000 claims abstract description 19
- 239000003112 inhibitor Substances 0.000 claims abstract description 19
- 229960001603 tamoxifen Drugs 0.000 claims abstract description 19
- 208000035199 Tetraploidy Diseases 0.000 claims abstract description 16
- 230000002706 hydrostatic effect Effects 0.000 claims abstract description 16
- 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 abstract description 15
- 229960003608 clomifene Drugs 0.000 claims abstract description 15
- 230000035939 shock Effects 0.000 claims abstract description 14
- 239000000463 material Substances 0.000 claims description 23
- 210000004602 germ cell Anatomy 0.000 claims description 18
- 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 16
- 238000011534 incubation Methods 0.000 claims description 12
- 235000013601 eggs Nutrition 0.000 claims description 10
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 229960003604 testosterone Drugs 0.000 claims description 8
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 8
- ZAMOUSCENKQFHK-UHFFFAOYSA-N Chlorine atom Chemical compound [Cl] ZAMOUSCENKQFHK-UHFFFAOYSA-N 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
- 230000027326 copulation Effects 0.000 claims description 4
- 230000002354 daily effect Effects 0.000 claims description 4
- 239000012153 distilled water Substances 0.000 claims description 4
- 230000008144 egg development Effects 0.000 claims description 4
- 230000003203 everyday effect Effects 0.000 claims description 4
- 230000000474 nursing effect Effects 0.000 claims description 4
- 230000002085 persistent effect Effects 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 235000009566 rice Nutrition 0.000 claims description 4
- 240000007594 Oryza sativa Species 0.000 claims 1
- 238000002791 soaking Methods 0.000 abstract description 2
- 241000251468 Actinopterygii Species 0.000 description 5
- 241000209094 Oryza Species 0.000 description 3
- 210000000349 chromosome Anatomy 0.000 description 2
- 238000003776 cleavage reaction Methods 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 230000001629 suppression Effects 0.000 description 2
- 230000004543 DNA replication Effects 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 206010020852 Hypertonia Diseases 0.000 description 1
- 208000020584 Polyploidy Diseases 0.000 description 1
- 230000036528 appetite Effects 0.000 description 1
- 235000019789 appetite Nutrition 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
- 238000009396 hybridization Methods 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 230000011278 mitosis Effects 0.000 description 1
- 239000008239 natural water Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 239000007787 solid Substances 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
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New or modified breeds of animals
- A01K67/027—New or modified breeds of vertebrates
- A01K67/0275—Genetically modified vertebrates, e.g. transgenic
-
- 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
- A01K2207/00—Modified animals
- A01K2207/25—Animals on a special diet
-
- 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
- A01K2217/00—Genetically modified animals
- A01K2217/03—Animals modified by random mutagenesis, e.g. using ENU, chemicals
-
- 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
- A01K2227/00—Animals characterised by species
- A01K2227/40—Fish
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Animal Behavior & Ethology (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Engineering & Computer Science (AREA)
- Feed For Specific Animals (AREA)
- Fodder In General (AREA)
Abstract
The invention discloses a method for increasing the breeding rate of a Channa argus triploid. The method comprises the following steps: preparing tamoxifen feed; feeding the Channa argus female parent with the tamoxifen feed; treating through cold shock and hydrostatic pressure; preparing an estrogen inhibitor solution; soaking a zygote in the estrogen inhibitor solution at normal temperature; feeding the fries with clomiphene larva feed; and fetching a male tetraploid as a male parent and a common diploid as a female parent, and backcrossing to obtain the triploid. Through the method, a 100% pure line of Channa argus triploid can be obtained, the breeding rate and emergence rate of the triploid are increased, the troubles in breeding and seed selection are reduced, and more importantly, the method is free of pollution to the germplasm of natural aquatic product variety and the ecological environment.
Description
Technical field
The present invention relates to the polyploid mating system of Ophicephalus argus, be specifically related to a kind of method improving Ophicephalus argus triploid incubation rate.
Background technology
Ophicephalus argus is commonly called as snakeheaded fish, and natural disposition is violent, and reproductive capacity is strong, and appetite is very big, and body colour is grey black, fast growth.The sterile feature of triploid Ophicephalus argus so that it is show significantly more advantage at aspects such as resistance, disease resistance and meats.In prior art, sterile triploid Ophicephalus argus is to be obtained with diploid interspecies crossing by tetraploid, but to obtain the triploid Ophicephalus argus of more than 98%, then be 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 mitosis or direct chromosome nonhomologous by method suppression germ cell such as hybridization doubles, prepare homology or allotrtraploid fish, wherein cold shock be usually with hydrostatic pressing process together with, 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 the tetraploid system that purity is high；Although and the method purity height hybridized, but success rate is low, the cycle is long.Additionally, tetraploid fish prepared by these methods above-mentioned, some can be educated, some can not be educated again, also likely to be present gene simultaneously and mixes phenomenon (allotetraploid), easily causes bio-hazard.
Summary of the invention
The deficiency existed for above-mentioned prior art, it is an object of the invention to provide a kind of method improving Ophicephalus argus triploid incubation rate that method is stable.
For reaching above-mentioned purpose, technical scheme is as follows:
A kind of method improving Ophicephalus argus triploid incubation rate, comprises the steps:
(1) tamoxifen feeds the outfit of material: takes the tamoxifen powder that purity is more than 95%, joins in Ophicephalus argus artificial breeding's feedstuff, makes nursing material, and wherein tamoxifen content is 15～20g/kg feedstuffs, stand-by；
(2) throw something and feed: before Ophicephalus argus female parent is laid eggs 5～10 days, routinely feedstuff feeding method Ophicephalus argus female parent is thrown something and fed tamoxifen feed material, twice daily, 8:00 in morning every 100g Ophicephalus argus female parent throws something and feeds 15～20g, and 6:00 in afternoon every 100g Ophicephalus argus female parent throws something and feeds 10～15g；Ripe to its egg development, then, take male Ophicephalus argus and put together so that it is natural spawning copulation is fertilized, and obtains germ cell, standby；
(3) cold shock+hydrostatic pressing processes: when a spilting of an egg, the method using cold shock to combine with hydrostatic pressing, and the germ cell in step (2) is put in the cold water that temperature is-1～1.0 DEG C, and hydrostatic pressing pressure is 180～300Kg/cm2, the persistent period is 2～3min；
(4) preparation of estrogen inhibitors solution: clomifene is dissolved in together with testosterone in the ethanol of 80%～90%, add distilled water again and be configured to estrogen inhibitors solution, the concentration that wherein estrogen inhibitors Chlorine in Solution rice is fragrant is 60～120mg/l, and the concentration of testosterone is 20～80mg/l；
(5) take the germ cell after step (3) processes and put in the estrogen inhibitors solution as prepared by step (4), soak at room temperature 10～30s, then take out and be put in normal incubation in hatchery, to emerging；
(6) fry in step (5) is taken, clomifene germling material of throwing something and feeding；Proportional quantity is that every kg germling material adds 10～15g clomifene, each feeding volume be every ten thousand tails 20～40g added with the germling material of clomifene, throw something and feed every day three times, until becoming Wu Zaimiao；Thus obtain male Ophicephalus argus tetraploid pure lines；
(7) triploid acquisition: take the male tetraploid of step (6) as male parent, takes common diploid as female parent, uses the mode backcrossed, it is thus achieved that 100% triploid.
Advantages of the present invention: 1, the present invention feeds tamoxifen to female parent, while playing suppression DNA replication dna and promoting male dual function, the germ cell First cleavage time is put off 3-5min, and improve the synchronization degree (up to 99%) of all germ cell First cleavages, for chromosome doubling with improve its tetraploid rate and provide the time and ensure and established solid foundation；2, use cold shock and relatively low hydrostatic pressing (less than conventional 350kg/cm in spilting of an egg of germ cell2), and the shorter process time (less than 5min), not only effectively reduce because cold shock overlong time, the hypertonia destructiveness to germ cell, improve the survival rate of germ cell, and reduce operation easier and pressurization cost；3, estrogen inhibitors solution soaking and clomifene germling material of throwing something and feeding, efficient hardening and the male conversion ratio of guarantee；4, backcross with original diploid female with the male tetraploid of pure lines, triploid purity has been effectively ensured.Thereby through the method, can obtain the Ophicephalus argus triploid pure lines of 100%, improve triploid incubation rate and emergence rate, decrease the trouble of breeding and seed selection, 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 Ophicephalus argus triploid incubation rate, comprises the steps:
(1) tamoxifen feeds the outfit of material: takes the tamoxifen powder that purity is more than 95%, joins in Ophicephalus argus artificial breeding's feedstuff, makes nursing material, and wherein tamoxifen content is 15g/kg feedstuff, stand-by；
(2) throwing something and feeding: before Ophicephalus argus female parent is laid eggs 5 days, feedstuff feeding method tamoxifen of throwing something and feeding Ophicephalus argus female parent feeds material routinely, and twice daily, 8:00 in morning every 100g Ophicephalus argus female parent 20g, 6:00 in afternoon every 100g Ophicephalus argus female parent of throwing something and feeding is thrown something and fed 15g；Ripe to its egg development, then, take male Ophicephalus argus and put together so that it is natural spawning copulation is fertilized, and obtains germ cell, standby；
(3) cold shock+hydrostatic pressing processes: when a spilting of an egg, the method using cold shock to combine with hydrostatic pressing, and the germ cell in step (2) is put in the cold water that temperature is 0 DEG C, and hydrostatic pressing pressure is 260Kg/cm2, the persistent period is 3min；
(4) preparation of estrogen inhibitors solution: clomifene is dissolved in the ethanol of 80% together with testosterone, then add distilled water and be configured to estrogen inhibitors solution, the concentration that wherein estrogen inhibitors Chlorine in Solution rice is fragrant is 60mg/l, and the concentration of testosterone is 20mg/l；
(5) take the germ cell after step (3) processes and put in the estrogen inhibitors solution as prepared by step (4), soak at room temperature 30s, then take out and be put in normal incubation in hatchery, to emerging；
(6) fry in step (5) is taken, clomifene germling material of throwing something and feeding；Proportional quantity is that every kg germling material adds 15g, and each feeding volume is every ten thousand tail 40g, throws something and feeds every day three times, until becoming Wu Zaimiao；Thus obtain male Ophicephalus argus tetraploid pure lines；
(7) triploid acquisition: take the male tetraploid of step (6) as male parent, takes common diploid as female parent, uses the mode backcrossed, it is thus achieved that 100% triploid.
Embodiment two
The method improving Ophicephalus argus triploid incubation rate, comprises the steps:
(1) tamoxifen feeds the outfit of material: takes the tamoxifen powder that purity is more than 95%, joins in Ophicephalus argus artificial breeding's feedstuff, makes nursing material, and wherein tamoxifen content is 20g/kg feedstuff, stand-by；
(2) throwing something and feeding: before Ophicephalus argus female parent is laid eggs 10 days, feedstuff feeding method tamoxifen of throwing something and feeding Ophicephalus argus female parent feeds material routinely, and twice daily, 8:00 in morning every 100g Ophicephalus argus female parent 20g, 6:00 in afternoon every 100g Ophicephalus argus female parent of throwing something and feeding is thrown something and fed 10g；Ripe to its egg development, then, take male Ophicephalus argus and put together so that it is natural spawning copulation is fertilized, and obtains germ cell, standby；
(3) cold shock+hydrostatic pressing processes: when a spilting of an egg, the method using cold shock to combine with hydrostatic pressing, and the germ cell in step (2) is put in the cold water that temperature is 0 DEG C, and hydrostatic pressing pressure is 300Kg/cm2, the persistent period is 3min；
(4) preparation of estrogen inhibitors solution: clomifene is dissolved in the ethanol of 90% together with testosterone, then add distilled water and be configured to estrogen inhibitors solution, the concentration that wherein estrogen inhibitors Chlorine in Solution rice is fragrant is 100mg/l, and the concentration of testosterone is 60mg/l；
(5) take the germ cell after step (3) processes and put in the estrogen inhibitors solution as prepared by step (4), soak at room temperature 10s, then take out and be put in normal incubation in hatchery, to emerging；
(6) fry in step (5) is taken, clomifene germling material of throwing something and feeding；Proportional quantity is that every kg germling material adds 10g, and each feeding volume is every ten thousand tail 20g, throws something and feeds every day three times, until becoming Wu Zaimiao；Thus obtain male Ophicephalus argus tetraploid pure lines；
(7) triploid acquisition: take the male tetraploid of step (6) as male parent, takes common diploid as female parent, uses the mode backcrossed, 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 is arbitrarily 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: Ophicephalus argus triploid is bred as result statistics
Note: the above results is 10 days tamoxifens of throwing something and feeding before Ophicephalus argus female parent is laid eggs, hydrostatic pressing processes 3 minutes, soak at room temperature 10s, every ten thousand tails throw something and feed the result acquired by clomifene 0.2g.
Claims (1)
1. the method improving Ophicephalus argus triploid incubation rate, comprises the steps:
(1) tamoxifen feeds the outfit of material: takes the tamoxifen powder that purity is more than 95%, joins in Ophicephalus argus artificial breeding's feedstuff, makes nursing material, and wherein tamoxifen content is 15～20g/kg feedstuffs, stand-by；
(2) throw something and feed: before Ophicephalus argus female parent is laid eggs 5～10 days, routinely feedstuff feeding method Ophicephalus argus female parent is thrown something and fed tamoxifen feed material, twice daily, 8:00 in morning every 100g Ophicephalus argus female parent throws something and feeds 15～20g, and 6:00 in afternoon every 100g Ophicephalus argus female parent throws something and feeds 10～15g；Ripe to its egg development, then, take male Ophicephalus argus and put together so that it is natural spawning copulation is fertilized, and obtains germ cell, standby；
(3) cold shock+hydrostatic pressing processes: when a spilting of an egg, the method using cold shock to combine with hydrostatic pressing, and the germ cell in step (2) is put in the cold water that temperature is-1～1.0 DEG C, and hydrostatic pressing pressure is 180～300Kg/cm2, the persistent period is 2～3min；
(4) preparation of estrogen inhibitors solution: clomifene is dissolved in together with testosterone in the ethanol of 80%～90%, add distilled water again and be configured to estrogen inhibitors solution, the concentration that wherein estrogen inhibitors Chlorine in Solution rice is fragrant is 60～120mg/l, and the concentration of testosterone is 20～80mg/l；
(5) take the germ cell after step (3) processes and put in the estrogen inhibitors solution as prepared by step (4), soak at room temperature 10～30s, then take out and be put in normal incubation in hatchery, to emerging；
(6) fry in step (5) is taken, clomifene germling material of throwing something and feeding；Proportional quantity is that every kg germling material adds 10～15g clomifene, each feeding volume be every ten thousand tails 20～40g added with the germling material of clomifene, throw something and feed every day three times, until becoming Wu Zaimiao；Thus obtain male Ophicephalus argus tetraploid pure lines；
(7) triploid acquisition: take the male tetraploid of step (6) as male parent, takes common diploid as female parent, uses the mode backcrossed, it is thus achieved that 100% triploid.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201610497626.7A CN105941332A (en) | 2016-06-30 | 2016-06-30 | Method for increasing breeding rate of Channa argus triploid |
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| CN201610497626.7A CN105941332A (en) | 2016-06-30 | 2016-06-30 | Method for increasing breeding rate of Channa argus triploid |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1957686A (en) * | 2006-11-20 | 2007-05-09 | 湖南文理学院 | Method of breeding pearls by using 6-dimethylin purine to induce triploid jibheader clams |
| CN102106292A (en) * | 2011-02-10 | 2011-06-29 | 中国水产科学研究院黄海水产研究所 | Flounder triploid batch producing method |
| CN102144600A (en) * | 2011-05-23 | 2011-08-10 | 浙江省海洋水产研究所 | Breeding method for inducing paralichthys olivaceus allotriploid by adopting physical method |
| CN103004653A (en) * | 2012-11-22 | 2013-04-03 | 中山大学 | Preparation method for triploid grouper |
| US20140200393A1 (en) * | 2013-01-15 | 2014-07-17 | Wisconsin Alumni Research Foundation | Methods and compositions for monitoring and enhancing early embryo development |
-
2016
- 2016-06-30 CN CN201610497626.7A patent/CN105941332A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1957686A (en) * | 2006-11-20 | 2007-05-09 | 湖南文理学院 | Method of breeding pearls by using 6-dimethylin purine to induce triploid jibheader clams |
| CN102106292A (en) * | 2011-02-10 | 2011-06-29 | 中国水产科学研究院黄海水产研究所 | Flounder triploid batch producing method |
| CN102144600A (en) * | 2011-05-23 | 2011-08-10 | 浙江省海洋水产研究所 | Breeding method for inducing paralichthys olivaceus allotriploid by adopting physical method |
| CN103004653A (en) * | 2012-11-22 | 2013-04-03 | 中山大学 | Preparation method for triploid grouper |
| US20140200393A1 (en) * | 2013-01-15 | 2014-07-17 | Wisconsin Alumni Research Foundation | Methods and compositions for monitoring and enhancing early embryo development |
Non-Patent Citations (3)
| Title |
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Application publication date: 20160921 |