CN105941330A - Efficient breeding method for male tetraploid pure lines of soft-shelled turtles - Google Patents
Efficient breeding method for male tetraploid pure lines of soft-shelled turtles Download PDFInfo
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- CN105941330A CN105941330A CN201610495560.8A CN201610495560A CN105941330A CN 105941330 A CN105941330 A CN 105941330A CN 201610495560 A CN201610495560 A CN 201610495560A CN 105941330 A CN105941330 A CN 105941330A
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- Prior art keywords
- trionyx sinensis
- sinensis wiegmann
- tamoxifen
- temperature
- soft
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- 208000035199 Tetraploidy Diseases 0.000 title claims abstract description 25
- 238000009395 breeding Methods 0.000 title claims abstract description 11
- 241001482311 Trionychidae Species 0.000 title 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
- 239000000463 material Substances 0.000 claims abstract description 20
- 229960001603 tamoxifen Drugs 0.000 claims abstract description 19
- 229940011871 estrogen Drugs 0.000 claims abstract description 18
- 239000000262 estrogen Substances 0.000 claims abstract description 18
- 239000003112 inhibitor Substances 0.000 claims abstract description 18
- 238000000034 method Methods 0.000 claims abstract description 14
- 241000736919 Pelodiscus sinensis Species 0.000 claims description 39
- 235000013601 eggs Nutrition 0.000 claims description 27
- 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
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 8
- 230000035939 shock Effects 0.000 claims description 7
- 229960003604 testosterone Drugs 0.000 claims description 6
- 230000004720 fertilization Effects 0.000 claims description 5
- 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
- 239000000443 aerosol Substances 0.000 claims description 4
- 229910052801 chlorine Inorganic materials 0.000 claims description 4
- 239000000460 chlorine Substances 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
- 230000003203 everyday effect Effects 0.000 claims description 4
- 230000000474 nursing 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
- 238000005507 spraying Methods 0.000 claims description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 4
- 101100279436 Caenorhabditis elegans egg-2 gene Proteins 0.000 claims description 2
- 238000004821 distillation Methods 0.000 claims description 2
- 150000002576 ketones Chemical class 0.000 claims description 2
- 210000001550 testis Anatomy 0.000 claims description 2
- 240000007594 Oryza sativa Species 0.000 claims 1
- 208000026487 Triploidy Diseases 0.000 abstract description 4
- 230000001488 breeding effect Effects 0.000 abstract 1
- 238000002635 electroconvulsive therapy Methods 0.000 abstract 1
- 241000270666 Testudines Species 0.000 description 9
- 241000209094 Oryza Species 0.000 description 3
- 238000003776 cleavage reaction Methods 0.000 description 2
- 239000012153 distilled water Substances 0.000 description 2
- 230000007017 scission Effects 0.000 description 2
- 241000270605 Chelonia Species 0.000 description 1
- 241000938605 Crocodylia Species 0.000 description 1
- 208000035240 Disease Resistance Diseases 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
- 230000007547 defect Effects 0.000 description 1
- 230000009977 dual effect Effects 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 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
- 210000004185 liver Anatomy 0.000 description 1
- 230000013011 mating Effects 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 208000011580 syndromic disease Diseases 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
- A01K2227/00—Animals characterised by species
- A01K2227/50—Amphibians, e.g. Xenopus
Landscapes
- 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)
- Fodder In General (AREA)
Abstract
The invention discloses an efficient breeding method for male tetraploid pure lines of soft-shelled turtles. The efficient breeding method comprises the following steps: preparing tamoxifen feeding materials, feeding female soft-shelled turtles with the tamoxifen feeding materials, carrying out heat shock treatment, preparing an estrogen inhibitor solution and obtaining the male tetraploid pure lines of the soft-shelled turtles. With the adoption of the method, the 100% male tetraploid pure lines of the soft-shelled turtles can be obtained; tetraploids of the soft-shelled turtles are effectively bred and the breeding rate and the yield of the tetraploids are improved; and a firm foundation is laid for the production of triploid sterile soft-shelled turtles.
Description
Technical field
The present invention relates to the polyploid mating system of Trionyx sinensis Wiegmann, be specifically related to the efficient breeding method of Trionyx sinensis Wiegmann male tetraploid pure lines.
Background technology
Trionyx sinensis Wiegmann, is excellent, also known as the soft-shelled turtle, Trionyx sinensis Wiegmann etc. with Trionyx sinensis (Wiegmann).It is subordinate to Reptilia, Chelonia, Trionychidae, common for two times
Body, Carnis Trionycis delicious flavour, nutritious, there are the effects such as clearing away heat and nourishing YIN, suppressing the hyperactive liver to relieve the wind syndrome, hard masses softening and resolving, diploid poor growth, support
The cycle of growing reaches 4-5.Because of triploid sterile feature, Trionyx sinensis Wiegmann is made to show the most aobvious at aspects such as the speed of growth, disease resistance, meats
The advantage write.In prior art, the sterile Trionyx sinensis Wiegmann of triploid to be obtained, method is by tetraploid Trionyx sinensis Wiegmann and diploid the most efficiently and effectively
Trionyx sinensis Wiegmann interspecies crossing obtains, then be accomplished by setting up the tetraploid Trionyx sinensis Wiegmann pure lines of inheritance stability, very high purity.But the genetic breeding Trionyx sinensis Wiegmann
In, up to the present yet there are no the report that tetraploid all-male Trionyx sinensis Wiegmann is efficiently bred as.
Summary of the invention
The defect existed for above-mentioned prior art, it is an object of the invention to provide that a kind of method is stable, population purity is high
The male tetraploid of turtle reaching 100% is sheerly efficient breeding method.
For reaching above-mentioned purpose, technical scheme is as follows:
The efficient breeding method of Trionyx sinensis Wiegmann male tetraploid pure lines, comprises the steps:
(1) tamoxifen feeds the outfit of material: takes the tamoxifen powder that purity is more than 95%, joins Trionyx sinensis Wiegmann routine and raise
In material, making nursing material, wherein tamoxifen content is 8～10g/500g feedstuffs, stand-by；
(2) throwing something and feeding: before female Trionyx sinensis Wiegmann is laid eggs, feedstuff feeding method tamoxifen of throwing something and feeding female Trionyx sinensis Wiegmann feeds material, every day routinely
Three times, 8:00 in morning every mother 100g Trionyx sinensis Wiegmann throws something and feeds 15～18g, and 12 noon every mother 100g Trionyx sinensis Wiegmann throws something and feeds 10～15g, and afternoon, 6:00 was every
Mother 100g Trionyx sinensis Wiegmann throws something and feeds 5～10g, until laying eggs；
(3) heat shock processes: fertile egg put into temperature be 25～32 DEG C, humidity be 40～80% constant-temperature incubator in, one
Before the secondary spilting of an egg 2～3min, temperature is quickly ramped up to 39～42 DEG C, keeps 3～5min；Then temperature is down to 28～32 DEG C；
(4) 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 180～250mg/l, testis
The concentration of ketone is 100～150mg/l；
(5) with the estrogen inhibitors solution prepared by step (4), put in aerosol apparatus, to the fertilization processed through step (3)
Egg carries out even spraying, and every 100 fertile eggs of consumption are 8～15ml, keep the temperature 28 of couveuse～32 DEG C, humidity 40～
80%, hatch；Thus obtain the male tetraploid pure lines of 100% Trionyx sinensis Wiegmann.
Advantages of the present invention: 1, in the present invention, female parent is added tamoxifen feedstuff feeding, plays suppression DNA fragmentation
While promote male dual function, by the time retardation of fertile egg First cleavage 3～5min, and improve all fertilizations
The synchronization extent (up to 99%) of egg First cleavage, ensures for operating next step offer time and establishes chromosome doubling basis；
2, use the heat shock short time to process fertile egg, not only effectively reduce the harmful effect to fertile egg, improve surviving of fertile egg
Rate；And further enhance tetraploid formation probability；3, estrogen inhibitors spray solution, strengthens again and ensures it
Male conversion ratio.Thereby through the method, the male tetraploid pure lines of the Trionyx sinensis Wiegmann of 100% can be obtained.The method, the most only effectively
It is bred as the tetraploid of Trionyx sinensis Wiegmann, and improves tetraploid incubation rate and emergence rate；Establish solid for producing the sterile Trionyx sinensis Wiegmann of triploid
Basis.
Detailed description of the invention
In conjunction with specific embodiment, the invention will be further described.
Embodiment one
The efficient breeding method of Trionyx sinensis Wiegmann male tetraploid pure lines, comprises the steps:
(1) tamoxifen feeds the outfit of material: take the tamoxifen powder that purity is more than 95%, joins turtle routine and raises
In feedstuff, making nursing material, wherein tamoxifen content is 10g/500g feedstuff, stand-by；
(2) throwing something and feeding: before female Trionyx sinensis Wiegmann is laid eggs, feedstuff feeding method tamoxifen of throwing something and feeding female parent feeds material, every day routinely
Three times, 8:00 in morning every 100g turtle throw something and feed 15g feed material, 12:00 at noon every 100g turtle throw something and feed 15g feed material, afternoon 6:
00 every 100g turtle throw something and feed 10g feed material, until laying eggs；
(3) heat shock processes: fertile egg put into temperature be 32 DEG C, humidity be 80% constant-temperature incubator in, before a spilting of an egg
3min, is quickly ramped up to 42 DEG C temperature, keeps 3min；Then temperature is down to 32 DEG C；
(4) 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
150mg/l；
(5) with the estrogen inhibitors solution prepared by step (4), put in aerosol apparatus, to the fertilization processed through step (3)
Egg carries out even spraying, and every 100 fertile eggs of consumption are 8ml, keeps the temperature 32 DEG C of couveuse, humidity 80%, hatches；
Thus obtain the male tetraploid pure lines of 100% Trionyx sinensis Wiegmann.
Embodiment two
The efficient breeding method of Trionyx sinensis Wiegmann male tetraploid pure lines, comprises the steps:
(1) tamoxifen feeds the outfit of material: take the tamoxifen powder that purity is more than 95%, joins turtle routine and raises
In feedstuff, making nursing material, wherein tamoxifen content is 8g/500g feedstuff, stand-by；
(2) throwing something and feeding: before female Trionyx sinensis Wiegmann is laid eggs, feedstuff feeding method tamoxifen of throwing something and feeding female parent feeds material, every day routinely
Three times, 8:00 in morning every 100g turtle throw something and feed 18g, 12:00 at noon every 100g turtle throw something and feed 15g feed material, afternoon, 6:00 was every
100g turtle is thrown something and fed 10g；
(3) heat shock processes: fertile egg put into temperature be 28 DEG C, humidity be 60% constant-temperature incubator in, before a spilting of an egg
3min, is quickly ramped up to 42 DEG C temperature, keeps 5min；Then temperature is down to 28 DEG C；
(4) preparation of estrogen inhibitors solution: clomifene is dissolved in the ethanol of 80% 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 230mg/l, and the concentration of testosterone is
140mg/l；
(5) with the estrogen inhibitors solution prepared by step (4), put in aerosol apparatus, to the fertilization processed through step (3)
Egg carries out even spraying, and every 100 fertile eggs of consumption are 15ml, keeps the humidity 60% of couveuse, temperature 28 DEG C, hatches；
Thus obtain the male tetraploid pure lines of 100% Trionyx sinensis Wiegmann.
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: the male tetraploid of Trionyx sinensis Wiegmann is bred as result statistics
Note: the above results is 3min, heat shock holding result acquired by 5min before a spilting of an egg, and wherein during heat shock, constant temperature is incubated
The temperature changing device is 28 DEG C, humidity is 60%.
Claims (1)
1. the efficient breeding method of Trionyx sinensis Wiegmann male tetraploid pure lines, comprises the steps:
(1) tamoxifen feeds the outfit of material: takes the tamoxifen powder that purity is more than 95%, joins Trionyx sinensis Wiegmann routine and raise
In material, making nursing material, wherein tamoxifen content is 8～10g/500g feedstuffs, stand-by；
(2) throwing something and feeding: before female Trionyx sinensis Wiegmann is laid eggs, feedstuff feeding method tamoxifen of throwing something and feeding female Trionyx sinensis Wiegmann feeds material, every day routinely
Three times, 8:00 in morning every mother 100g Trionyx sinensis Wiegmann throws something and feeds 15～18g, and 12 noon every mother 100g Trionyx sinensis Wiegmann throws something and feeds 10～15g, and afternoon, 6:00 was every
Mother 100g Trionyx sinensis Wiegmann throws something and feeds 5～10g, until laying eggs；
(3) heat shock processes: fertile egg put into temperature be 25～32 DEG C, humidity be 40～80% constant-temperature incubator in, one
Before the secondary spilting of an egg 2～3min, temperature is quickly ramped up to 39～42 DEG C, keeps 3～5min；Then temperature is down to 28～32 DEG C；
(4) 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 180～250mg/l, testis
The concentration of ketone is 100～150mg/l；
(5) with the estrogen inhibitors solution prepared by step (4), put in aerosol apparatus, to the fertilization processed through step (3)
Egg carries out even spraying, and every 100 fertile eggs of consumption are 8～15ml, keep the temperature 28 of couveuse～32 DEG C, humidity 40～
80%, hatch；Thus obtain the male tetraploid pure lines of 100% Trionyx sinensis Wiegmann.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610495560.8A CN105941330A (en) | 2016-06-30 | 2016-06-30 | Efficient breeding method for male tetraploid pure lines of soft-shelled turtles |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610495560.8A CN105941330A (en) | 2016-06-30 | 2016-06-30 | Efficient breeding method for male tetraploid pure lines of soft-shelled turtles |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105941330A true CN105941330A (en) | 2016-09-21 |
Family
ID=56901690
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610495560.8A Pending CN105941330A (en) | 2016-06-30 | 2016-06-30 | Efficient breeding method for male tetraploid pure lines of soft-shelled turtles |
Country Status (1)
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1861017A (en) * | 2005-10-28 | 2006-11-15 | 白宝海 | Heat-shock and static water-pressure type method for prodn. of tetraploid of trout |
| CN1969616A (en) * | 2005-11-25 | 2007-05-30 | 李思发 | Method for constructing allotetraploid of bluntsnout bream |
| WO2008132349A2 (en) * | 2007-03-23 | 2008-11-06 | Institut Francais De La Recherche Pour L'exploitation De La Mer (Ifremer) | Production of tetraploid bivalve molluscs from crossing diploid females and tetraploid males |
| CN103960175A (en) * | 2013-01-31 | 2014-08-06 | 中国科学院海洋研究所 | Method for batch induction of scophthalmus maximus tetraploid fries |
| CN104322408A (en) * | 2014-10-17 | 2015-02-04 | 中山大学 | Siniperca chuatsi polyploid induction method |
-
2016
- 2016-06-30 CN CN201610495560.8A patent/CN105941330A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1861017A (en) * | 2005-10-28 | 2006-11-15 | 白宝海 | Heat-shock and static water-pressure type method for prodn. of tetraploid of trout |
| CN1969616A (en) * | 2005-11-25 | 2007-05-30 | 李思发 | Method for constructing allotetraploid of bluntsnout bream |
| WO2008132349A2 (en) * | 2007-03-23 | 2008-11-06 | Institut Francais De La Recherche Pour L'exploitation De La Mer (Ifremer) | Production of tetraploid bivalve molluscs from crossing diploid females and tetraploid males |
| CN103960175A (en) * | 2013-01-31 | 2014-08-06 | 中国科学院海洋研究所 | Method for batch induction of scophthalmus maximus tetraploid fries |
| CN104322408A (en) * | 2014-10-17 | 2015-02-04 | 中山大学 | Siniperca chuatsi polyploid induction method |
Non-Patent Citations (3)
| Title |
|---|
| 南平 等: "人工诱导四倍体泥鳅的研究", 《河南师范大学学报(自然科学版)》 * |
| 汤光 主编: "《实用内科药物手册》", 31 January 2001, 中国医药科技出版社 * |
| 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 农业科技辑》 * |
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Application publication date: 20160921 |