CN105941331A - Efficient breeding method for triploid pure lines of snails - Google Patents
Efficient breeding method for triploid pure lines of snails Download PDFInfo
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- CN105941331A CN105941331A CN201610496054.0A CN201610496054A CN105941331A CN 105941331 A CN105941331 A CN 105941331A CN 201610496054 A CN201610496054 A CN 201610496054A CN 105941331 A CN105941331 A CN 105941331A
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- spiral shell
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- 208000026487 Triploidy Diseases 0.000 title claims abstract description 19
- 238000009395 breeding Methods 0.000 title claims abstract description 18
- 241000237858 Gastropoda Species 0.000 title abstract description 7
- 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
- 238000000034 method Methods 0.000 claims abstract description 25
- 229940011871 estrogen Drugs 0.000 claims abstract description 20
- 239000000262 estrogen Substances 0.000 claims abstract description 20
- 239000003112 inhibitor Substances 0.000 claims abstract description 20
- 229960001603 tamoxifen Drugs 0.000 claims abstract description 19
- 208000035199 Tetraploidy Diseases 0.000 claims abstract description 18
- 239000000463 material Substances 0.000 claims abstract description 18
- 230000001488 breeding effect Effects 0.000 claims abstract description 7
- 102000002322 Egg Proteins Human genes 0.000 claims description 20
- 108010000912 Egg Proteins Proteins 0.000 claims description 20
- 210000000481 breast Anatomy 0.000 claims description 20
- 210000004681 ovum Anatomy 0.000 claims description 20
- 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
- 235000013601 eggs Nutrition 0.000 claims description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 230000004720 fertilization Effects 0.000 claims description 8
- 229960003604 testosterone Drugs 0.000 claims description 8
- 230000000474 nursing effect Effects 0.000 claims description 6
- 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
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims description 4
- 238000003287 bathing Methods 0.000 claims description 4
- 239000000460 chlorine Substances 0.000 claims description 4
- 229910052801 chlorine Inorganic materials 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
- 239000012153 distilled water Substances 0.000 claims description 4
- 230000008144 egg development Effects 0.000 claims description 4
- 239000001301 oxygen Substances 0.000 claims description 4
- 229910052760 oxygen Inorganic materials 0.000 claims description 4
- 238000006213 oxygenation reaction Methods 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
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- 240000007594 Oryza sativa Species 0.000 claims 1
- 239000006260 foam Substances 0.000 abstract 1
- 230000003068 static effect Effects 0.000 abstract 1
- 210000004602 germ cell Anatomy 0.000 description 5
- 241000209094 Oryza Species 0.000 description 3
- 238000003776 cleavage reaction Methods 0.000 description 3
- 230000007017 scission Effects 0.000 description 3
- 238000002791 soaking Methods 0.000 description 2
- 241000984614 Bellamya Species 0.000 description 1
- 241001094455 Cipangopaludina chinensis Species 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 208000020584 Polyploidy Diseases 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- 238000012214 genetic breeding Methods 0.000 description 1
- 230000009931 harmful effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000008239 natural water Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 230000001629 suppression Effects 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
- A01K2227/70—Invertebrates
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Animal Behavior & Ethology (AREA)
- Zoology (AREA)
- Animal Husbandry (AREA)
- Biodiversity & Conservation Biology (AREA)
- Farming Of Fish And Shellfish (AREA)
Abstract
The invention discloses an efficient breeding method for triploid pure lines of snails. The efficient breeding method comprises the following steps: preparing tamoxifen feeding materials, feeding female snails with the tamoxifen feeding materials, carrying out static air pressure treatment, preparing an estrogen inhibitor solution, preparing an estrogen inhibitor solution foam bath, obtaining male tetraploid pure lines of the snails and obtaining triploids by adopting a backcrossing manner. With the adoption of the method, the 100% triploid pure lines can be obtained; the breeding rate and the yield of the tetraploids are improved and troubles of breeding and selecting strains are reduced; and more importantly, the efficient breeding method has no pollution to variety and germplasm of natural aquatic products and the ecological environment.
Description
Technical field
The present invention relates to the polyploid mating system of spiral shell, be specifically related to spiral shell triploid and be sheerly efficient breeding method.
Background technology
The distribution of spiral shell, kind are many; refer to gastropod (gastropod) animal, particularly relate to a shell closed, can be entirely retracted to wherein with the gastropod that must protect; common are Bellamya sp, Cipangopaludina chinensis etc., the mode of reproduction of spiral shell is generally ovoviviparity.Because of the triploid sterile feature of spiral shell so that it is show significantly more advantage at aspects such as the speed of growth, disease resistance, meats.In prior art, the sterile spiral shell of triploid to be obtained, method is to be obtained with diploid spiral shell interspecies crossing by tetraploid spiral shell the most efficiently and effectively, then be accomplished by setting up the tetraploid spiral shell pure lines of inheritance stability, very high purity.But in the genetic breeding of spiral shell, up to the present yet there are no the report that tetraploid all-male spiral shell is bred as, more have no the precedent backcrossed with diploid female parent with all-male tetraploid.
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 spiral shell triploid stable, population high purity 100% and is sheerly efficient breeding method.
For reaching above-mentioned purpose, technical scheme is as follows:
Spiral shell triploid is sheerly efficient breeding method, comprises the steps:
(1) tamoxifen feeds the outfit of material: takes the tamoxifen powder that purity is more than 90-95%, joins in spiral shell artificial breeding's feedstuff, makes nursing material, and wherein tamoxifen content is 10～15g/500g nursing material, stand-by；
(2) throwing something and feeding: before female spiral shell is laid eggs 5～10 days, feedstuff feeding method tamoxifen of throwing something and feeding female spiral shell feeds material routinely, and twice daily, 8:00 in morning every mother 100g spiral shell throws something and feeds 5～10g, and 6:00 in afternoon every mother 100g spiral shell throws something and feeds 10～15g；Ripe to its egg development, then, take public spiral shell and put together so that it is natural spawning fertilization, bosom ovum mother's spiral shell of must being fertilized；
(3) quiet pneumatics processes: when a spilting of an egg, the method using quiet pneumatics, and the fertilization bosom ovum mother's spiral shell in step (2) is put in the quiet pneumatics case that temperature is normal temperature laboratory, and quiet pneumatics pressure is 250～340Kg/cm2, the persistent period is 3～5min；
(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 100～200mg/l, and the concentration of testosterone is 10～50mg/l；
(5) taking the bosom ovum mother's spiral shell after step (3) processes, put into in the estrogen inhibitors solution prepared by step (4), room temperature bathing 60～120S, bosom ovum mother's spiral shell and solution ratio for 1:2, by oxygenation, makes the oxygen content in solution reach 5～10mg/l；
(6) take the bosom ovum mother's spiral shell after step (5) processes, put into and nursery pond carries out conventional seedbed system cultivation；Until output children's spiral shell, thus obtain the male tetraploid pure lines of spiral shell；
(7) take the male tetraploid of step (6) as male parent, take common diploid as female parent, use the mode backcrossed, it is thus achieved that 100% triploid.
Advantages of the present invention: 1, in the present invention, female parent is added tamoxifen feedstuff feeding, while playing suppression DNA fragmentation and promoting male dual function, by the time retardation of germ cell First cleavage 3～5min, and improve the synchronization extent (up to 99%) of all germ cell First cleavages, ensure for operating next step offer time and establish chromosome doubling basis；2, use relatively low quiet pneumatics pressure (less than conventional 350kg/cm at germ cell First cleavage2) and shorter process time (less than 5min), not only effectively reduce the harmful effect to germ cell, improve the survival rate of germ cell；And further enhance tetraploid formation probability；3, estrogen inhibitors solution soaking, strengthens again and ensures its male conversion ratio；4, backcross with original diploid female with the male tetraploid of pure lines, triploid purity has been effectively ensured.Thereby through the method, the triploid pure lines of 100% can be obtained, improve triploid incubation rate and emergence rate, decrease the trouble of breeding and seed selection, be more mainly matter all to Natural Water product and ecological environment is pollution-free.
Detailed description of the invention
In conjunction with specific embodiment, the invention will be further described.
Embodiment one
The male tetraploid of spiral shell is sheerly efficient breeding method, comprises the steps:
(1) tamoxifen feeds the outfit of material: takes the tamoxifen powder that purity is more than 95%, joins in spiral shell artificial breeding's feedstuff, makes nursing material, and wherein tamoxifen content is that 10g/500g feeds material, stand-by；
(2) throwing something and feeding: before female spiral shell is laid eggs 5 days, feedstuff feeding method tamoxifen of throwing something and feeding female spiral shell feeds material routinely, and twice daily, 8:00 in morning every mother 100g spiral shell 10g, 6:00 in afternoon every mother 100g spiral shell of throwing something and feeding is thrown something and fed 15g；Ripe to its egg development, then, take public spiral shell and put together so that it is natural spawning fertilization, bosom ovum mother's spiral shell of must being fertilized；
(3) quiet pneumatics processes: when a spilting of an egg, the method using quiet pneumatics, and the fertilization bosom ovum mother's spiral shell in step (2) is put in the quiet pneumatics case that temperature is normal temperature laboratory, and quiet pneumatics pressure is 340Kg/cm2, the persistent period is 3min；
(4) estrogen inhibitors preparation: 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 200mg/l, and the concentration of testosterone is 50mg/l；
(5) taking the bosom ovum mother's spiral shell after step (3) processes, put into in the estrogen inhibitors solution prepared by step (4), room temperature bathing 120S, bosom ovum mother's spiral shell and solution ratio for 1:2, by oxygenation, makes the oxygen content in solution reach 5mg/l；
(6) take the bosom ovum mother's spiral shell after step (5) processes, put into and nursery pond carries out conventional seedbed system cultivation；Until output children's spiral shell, thus obtain the male tetraploid pure lines of spiral shell.
(7) take the male tetraploid of step (6) as male parent, take common diploid as female parent, use the mode backcrossed, it is thus achieved that 100% triploid.
Embodiment two
The male tetraploid of spiral shell is sheerly efficient breeding method, comprises the steps:
(1) tamoxifen feeds the outfit of material: takes the tamoxifen powder that purity is more than 95%, joins in spiral shell artificial breeding's feedstuff, makes nursing material, and wherein tamoxifen content is that 15g/500g feeds material, stand-by；
(2) throwing something and feeding: before female spiral shell is laid eggs 10 days, feedstuff feeding method tamoxifen of throwing something and feeding female spiral shell feeds material routinely, and twice daily, 8:00 in morning every mother 100g spiral shell throws something and feeds 5～10g, and 6:00 in afternoon every mother 100g spiral shell is thrown something and fed 10g；Ripe to its egg development, then, take public spiral shell and put together so that it is natural spawning fertilization, bosom ovum mother's spiral shell of must being fertilized；
(3) quiet pneumatics processes: when a spilting of an egg, the method using quiet pneumatics, and the fertilization bosom ovum mother's spiral shell in step (2) is put in the quiet pneumatics case that temperature is normal temperature laboratory, and quiet pneumatics pressure is 250Kg/cm2, the persistent period is 5min；
(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 180mg/l, and the concentration of testosterone is 30mg/l；
(5) taking the bosom ovum mother's spiral shell after step (3) processes, put into in the estrogen inhibitors solution prepared by step (4), room temperature bathing 60S, bosom ovum mother's spiral shell and solution ratio for 1:2, by oxygenation, makes the oxygen content in solution reach 10mg/l；
(6) take the bosom ovum mother's spiral shell after step (5) processes, put into and nursery pond carries out conventional seedbed system cultivation；Until output children's spiral shell, thus obtain the male tetraploid pure lines of spiral shell.
(7) take the male tetraploid of step (6) as male parent, take common diploid as female parent, use 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: spiral shell triploid is bred as result statistics
Note: the above results is that female spiral shell is laid eggs first 10 days tamoxifens of throwing something and feeding, and the quiet pneumatics of cold shock keeps the result acquired by 5min, estrogen inhibitors solution soaking 60s.
Claims (1)
1. spiral shell triploid is sheerly efficient breeding method, comprises the steps:
(1) tamoxifen feeds the outfit of material: takes the tamoxifen powder that purity is more than 90-95%, joins in spiral shell artificial breeding's feedstuff, makes nursing material, and wherein tamoxifen content is 10～15g/500g nursing material, stand-by；
(2) throwing something and feeding: before female spiral shell is laid eggs 5～10 days, feedstuff feeding method tamoxifen of throwing something and feeding female spiral shell feeds material routinely, and twice daily, 8:00 in morning every mother 100g spiral shell throws something and feeds 5～10g, and 6:00 in afternoon every mother 100g spiral shell throws something and feeds 10～15g；Ripe to its egg development, then, take public spiral shell and put together so that it is natural spawning fertilization, bosom ovum mother's spiral shell of must being fertilized；
(3) quiet pneumatics processes: when a spilting of an egg, the method using quiet pneumatics, and the fertilization bosom ovum mother's spiral shell in step (2) is put in the quiet pneumatics case that temperature is normal temperature laboratory, and quiet pneumatics pressure is 250～340Kg/cm2, the persistent period is 3～5min；
(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 100～200mg/l, and the concentration of testosterone is 10～50mg/l；
(5) taking the bosom ovum mother's spiral shell after step (3) processes, put into in the estrogen inhibitors solution prepared by step (4), room temperature bathing 60～120S, bosom ovum mother's spiral shell and solution ratio for 1:2, by oxygenation, makes the oxygen content in solution reach 5～10mg/l；
(6) take the bosom ovum mother's spiral shell after step (5) processes, put into and nursery pond carries out conventional seedbed system cultivation；Until output children's spiral shell, thus obtain the male tetraploid pure lines of spiral shell；
(7) take the male tetraploid of step (6) as male parent, take common diploid as female parent, use the mode backcrossed, it is thus achieved that 100% triploid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610496054.0A CN105941331A (en) | 2016-06-30 | 2016-06-30 | Efficient breeding method for triploid pure lines of snails |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610496054.0A CN105941331A (en) | 2016-06-30 | 2016-06-30 | Efficient breeding method for triploid pure lines of snails |
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| Publication Number | Publication Date |
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| CN105941331A true CN105941331A (en) | 2016-09-21 |
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|---|---|---|---|
| CN201610496054.0A Pending CN105941331A (en) | 2016-06-30 | 2016-06-30 | Efficient breeding method for triploid pure lines of snails |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102388838A (en) * | 2011-09-13 | 2012-03-28 | 上海博琛生物科技有限公司 | Method and device for improving male hatching rate of oviparity |
| CN102988986A (en) * | 2012-11-02 | 2013-03-27 | 浙江万里学院 | Male trait inducer and induction method for pelodiscus sinensis in embryonic periods |
| CN103243068A (en) * | 2013-04-27 | 2013-08-14 | 上海交通大学 | Method for rapidly evaluating composition effect of estrogen and antiestrogen |
| CN105409873A (en) * | 2015-11-13 | 2016-03-23 | 丽水学院 | Method for controlling hoplobatrachus tigerinus sex ratio and application of method |
-
2016
- 2016-06-30 CN CN201610496054.0A patent/CN105941331A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102388838A (en) * | 2011-09-13 | 2012-03-28 | 上海博琛生物科技有限公司 | Method and device for improving male hatching rate of oviparity |
| CN102988986A (en) * | 2012-11-02 | 2013-03-27 | 浙江万里学院 | Male trait inducer and induction method for pelodiscus sinensis in embryonic periods |
| CN103243068A (en) * | 2013-04-27 | 2013-08-14 | 上海交通大学 | Method for rapidly evaluating composition effect of estrogen and antiestrogen |
| CN105409873A (en) * | 2015-11-13 | 2016-03-23 | 丽水学院 | Method for controlling hoplobatrachus tigerinus sex ratio and application of method |
Non-Patent Citations (4)
| Title |
|---|
| 李晓诗等: "他莫昔芬的药理作用机制及其对卵巢功能的影响", 《中国普外基础与临床杂志》 * |
| 桂建芳等: "鱼类染色体组操作的研究Ⅱ静水压处理和静水压与冷休克结合处理诱导水晶彩鲫四倍体", 《水生生物学报》 * |
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| 范鹏: "甲基睾酮(MT)对雌性及未性分化斑马鱼(Danio rerio)的影响", 《中国优秀硕士学位论文全文数据库》 * |
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