CN104585083A - Method for distant hybridization between subfamilies of cyprinus carpio and megalobrama amblycephala - Google Patents
Method for distant hybridization between subfamilies of cyprinus carpio and megalobrama amblycephala Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; CARE OF BIRDS, FISHES, INSECTS; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
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Abstract
A method for distant hybridization between subfamilies of cyprinus carpio and megalobrama amblycephala includes the following steps that firstly, female cyprinus carpio and male megalobrama amblycephala which have obvious sexual maturity characteristics and good somatic features are selected as parent fishes for special pool breeding, then, artificial spawning induction is conducted on the parent finishes in a breeding season, the parent fishes with a good spawning induction effect are selected to be subjected to artificial dry-method fertilization, running water incubation is conducted on fertilized embryos in a pure water incubation device, fish fry are raised after being incubated, the raised fish fry are detected and screened, hybrid offspring of the cyprinus carpio and the megalobrama amblycephala are obtained through flow cytometry DNA content determination and renal tissue chromosome ploidy detection, and the hybrid offspring comprise diploid natural gynogenesis cyprinus carpio, diploid natural gynogenesis mirror carps, diploid fish like the cyprinus carpio and tetraploid hybrid fish. The method has the advantages that the rate of fertilization is high, the incubating rate is high, the offspring are excellent in character, and the method has great significance in genetic breeding.
Description
Technical field
The present invention relates to fish hybridization method, particularly relate to the distant hybridization method in cyprinid fish between subfamily.
Background technology
Assay affiliation between planting or between planting above hybridization be called distant hybridization, be the method that fish character improvement is commonly used.Distant hybridization is as a kind of important genetics-breeding in fish and character improvement method, it can integrate a whole set of foreign gene, thus change the expression regulation of filial generation gene, make filial generation may show hybrid vigour in growth rate, resistance, disease resistance and meat etc.Distant hybridization, according to different parental combinations, can be formed in dissimilar fishes all different in phenotype and genotype.F in the filial generation that inter-genera distant hybridization as red crucian carp and Xiangjiang wild carps is formed
1-F
2for diploid hybrid fish (2n=100), F
3-F
24for allotrtraploid fish (4n=200); Utilize allotrtraploid fish and relevant liploid fish mating, define the triploid fish (3n=150) with advantages such as fast growth, Fresh & Tender in Texture, premunition is strong on a large scale.The F of the inter-subfamily distant hybridization of red crucian carp and megalobrama amblycephala
1in define the red crucian carp of dliploid Natural Gynogenesis (2n=100), allotetraploid crucian carp triangular bream (4n=148) and allotriploid crucian carp triangular bream (3n=124); Wherein allotetraploid crucian carp triangular bream defines autotetraploid fish products system (4n=200) by selfing.
Carp (Cyprinus carpio carpio) belongs to the liploid fish (2n=100) in Cyprinidae, carp subfamily, and megalobrama amblycephala (Megalobrama amblycephala) belongs to the liploid fish (2n=48) in Cyprinidae, Culter subfamily; In the biological properties such as heredity, classification and the bodily form, there is larger difference between carp and megalobrama amblycephala, especially between them, chromosome number of somatic is widely different, and the chromosome number of carp is 100, and the chromosome number of megalobrama amblycephala is 48, and not sexual intercourse at double.So far also do not have carp and megalobrame amblycephala subfamily distant hydridization successfully to report both at home and abroad, explore and a kind ofly make the cross method between carp and megalobrama amblycephala and the biological property of its offspring is researched and analysed that there is important using value.
Summary of the invention
Technical problem to be solved by this invention is, overcome the deficiency and defect mentioned in above background technology, provide that a kind of fertilization rate is high, incubation rate is high, characters of progenies is excellent, method to the significant carp of genetic breeding and megalobrame amblycephala subfamily distant hydridization.
For solving the problems of the technologies described above, the technical scheme that the present invention proposes is a kind of method of carp and megalobrame amblycephala subfamily distant hydridization, comprise the following steps: first select sexual maturity feature obvious, the female carp that sign is good and male megalobrama amblycephala are cultivated as the special pond of parent population, then in mating season, artificial induced spawning is carried out to described parent population, select the good parent population of spawning effect and carry out artificial dry method insemination, embryo after having inseminated carries out Lotic hatching in water purification hatching apparatus, raise after fry hatching completes, again the fry after raising is detected, screening, measured by fluidic cell DNA content, nephridial tissue ploidy detects and obtains carp and megalobrama amblycephala filial generation, filial generation comprises dliploid Natural Gynogenesis carp, dliploid Natural Gynogenesis mirror carp, the liploid fish of class crucian and Tetraploid fish.The hybridization fish of the Four types adopting preceding solution to obtain all has the advantages such as bodily form grace, fast growth, strong stress resistance, in novel excellent fish strain development and genetic breeding, have important value.
Above-mentioned carp and the method for megalobrame amblycephala subfamily distant hydridization, preferably, described method specifically comprises the following steps:
A the selection of () parent population and cultivation: before breeding period 3 ~ 4 months, select sexual maturity feature is obvious, sign is good female carp and male megalobrama amblycephala and carry out the cultivation of special pond respectively as the maternal parent population of distant hybridization and male parent parent population, keep water quality good; Raise with smart bait about the breeding period previous moon, every 2 ~ 3 days stimulation by running water once, promote parent population gonadal maturation; The good parent population of sign generally shows as that build is good, body colour is bright-coloured, physique is healthy and strong, anosis without wound;
(b) artificial induced spawning: in mid-May then to early June, when water temperature stability is more than 20 DEG C, for described parent population injection Luteinizing hormone releasing hormone analog (LRH-A) is hastened parturition with the ocyodinic that mixes of human chorionic gonadtropin (HCG), the consumption of described Luteinizing hormone releasing hormone analog is 6 ~ 10 μ g/kg (most preferably 10 μ g/kg), and the consumption of human chorionic gonadtropin is 500 ~ 800IU/kg (most preferably 600IU/kg); First inject maternal parent population, inject male parent parent population again after 4 ~ 5h, the injected dose of male parent parent population reduces by half; By the number ratio of 1: 6 ~ 8, maternal parent population, male parent parent population are put into same spawning pond after injection, then wash by water in pond in time depending on regimen, especially 3 ~ 4h before laying eggs, after injecting certain water yield, stop water filling, allow fish hasten parturition in hydrostatic, until it starts to lay eggs and produce essence smoothly; Preferably adopt pectoral fin base portion without abdominal cavity one, squama place pin injection during injection, to improve the survival rate of parent population;
C () insemination hatching: select the maternal parent population smoothly that lays eggs, fix with canvas stretcher, so that operation and the damage of minimizing to maternal parent population; Then measure large male parent parent population carry out artificial dry method insemination with product essence, it is that the water purification hatching apparatus of 20 DEG C ~ 21 DEG C carries out Lotic hatching that fertilized egg is placed in water temperature;
D () raises: after fry all hatches, continue cultivation 2 ~ 3 days in incubator, and occur being proceeded to by fry after waist point in the pond of fostering and apply fertilizer in advance until fry and raise, soya-bean milk of splashing after 1 ~ 2 day, makes every effort to thin, even by 2 ~ 3 times/day; Detect obtain filial generation by fluidic cell DNA content mensuration, nephridial tissue ploidy after fry grows up to.The chromosome map of Four types filial generation is respectively as shown in Fig. 4, Fig. 6, Fig. 8 and Figure 10.
In the present invention, fluidic cell DNA content determination method step is: to take a blood sample about 0.2mL from fish tail vein blood vessel with through the wetting disposable syringe of heparin, inject the Eppendorf pipe that 0.8% physiological saline is housed, 1mL nucleus extraction liquid DAPI-A (nuclei extraction solution is added in blood and mixed liquor of normal saline, Germany Partec Gmbh provides), processing time 10 ~ 15min; Sample filters through 20 μm of NFs (German Partec GmbH provides); DNA dyeing liquor (DAPI-B, German Partec GmbH provides) is statically placed in dark place stained specimens about 5 ~ 10min, then goes up machine testing.By fluidic cell DNA content measure can be clear and definite the ratio knowing dliploid Natural Gynogenesis carp in carp and megalobrama amblycephala filial generation, dliploid Natural Gynogenesis mirror carp, the liploid fish of class crucian and the DNA average content of this Four types filial generation of Tetraploid fish and the DNA average content with the common red crucian carp of object of reference.
The detection method of middle kidney tissue staining body ploidy of the present invention comprises: under 18 DEG C ~ 22 DEG C water temperatures, cultural aim fish is after 1 ~ 3 day, PHA is injected 1 ~ 3 time to experiment fish, each dosage is 2 ~ 8 μ g/g body weight, interval time is 12 ~ 24 hours, before dissection is drawn materials 2 ~ 6 hours, injection colchicine, dosage is 2 ~ 4 μ g/g body weight.Take out nephridial tissue, under physiological saline, shred nephridial tissue, 0.075mol/L KCL Hypotonic treatment, at 20 DEG C of temperature hypotonic 40 ~ 60 minutes; With glacial acetic acid: the fixing nephridial tissue of methyl alcohol (1: 3) 1 ~ 3 time; Freezing slide drips sheet; Giemsa dye liquor dyeing 45 ~ 60 minutes, thin current rinse the slide back side and remove dye liquor, air-dry rear microscopy, take pictures.Detected by nephridial tissue ploidy and also clearly can know dliploid Natural Gynogenesis carp in carp and megalobrama amblycephala filial generation, dliploid Natural Gynogenesis mirror carp, the liploid fish of class crucian and the chromosome number of this Four types of Tetraploid fish and ploidy thereof.
Compared with prior art, the invention has the advantages that: in the present invention, the F that the inter-subfamily distant hybridization of carp and megalobrama amblycephala is formed
1in define dliploid Natural Gynogenesis carp (2n=100), the liploid fish (2n=100) of dliploid Natural Gynogenesis mirror carp (2n=100) and class crucian and Tetraploid fish (4n=148), these filial generations have the advantages such as bodily form grace, fast growth, strong stress resistance, present the distinguishing feature of bio-diversity simultaneously, in novel excellent fish strain development and genetic breeding, there is important value.Filial generation in the present invention is obviously with the parent of existing Distant crossing combination with hybridize and produce offspring and there is marked difference, it is high that method of the present invention has fertilization rate, incubation rate is high, the advantages such as characters of progenies is excellent, the dliploid Natural Gynogenesis carp that Distant crossing combination of the present invention obtains, dliploid Natural Gynogenesis mirror carp, the liploid fish of class crucian and Tetraploid fish be expected to after hybridization generation in cultivate dliploid Natural Gynogenesis carp strain by hereditary and selection, dliploid Natural Gynogenesis mirror carp strain, the liploid fish strain of class crucian and Tetraploid fish products system, wherein by continuing seed selection to Tetraploid fish products system, be expected to develop novel autotetraploid fish products system.The present invention has important biological significance in organic evolution and genetics-breeding in fish.
Accompanying drawing explanation
In order to be illustrated more clearly in the embodiment of the present invention or technical scheme of the prior art, be briefly described to the accompanying drawing used required in embodiment or description of the prior art below, apparently, accompanying drawing in the following describes is some embodiments of the present invention, for those of ordinary skill in the art, under the prerequisite not paying creative work, other accompanying drawing can also be obtained according to these accompanying drawings.
Fig. 1 is the profile photo of carp (♀) in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) parent.
Fig. 2 is the profile photo of megalobrama amblycephala (♂) in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) parent.
Fig. 3 is the profile photo of dliploid Natural Gynogenesis carp in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) offspring.
Fig. 4 is the chromosome map (2n=100) of dliploid Natural Gynogenesis carp in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) offspring.
Fig. 5 is the profile photo of dliploid Natural Gynogenesis mirror carp in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) offspring.
Fig. 6 is the chromosome map (2n=100) of dliploid Natural Gynogenesis mirror carp in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) offspring.
Fig. 7 is the profile photo of the liploid fish of class crucian in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) offspring.
Fig. 8 is the chromosome map (2n=100) of the liploid fish of class crucian in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) offspring.
Fig. 9 is the profile photo of Tetraploid fish in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) offspring.
Figure 10 is the chromosome map (4n=148) of Tetraploid fish in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) offspring.
Figure 11 is the DNA content figure of the common red crucian carp of object of reference in the embodiment of the present invention.
Figure 12 is the DNA content figure of dliploid Natural Gynogenesis carp in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) offspring.
Figure 13 is the DNA content figure of dliploid Natural Gynogenesis mirror carp in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) offspring.
Figure 14 is the DNA content figure of the liploid fish of class crucian in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) offspring.
Figure 15 is the DNA content figure of Tetraploid fish in carp (♀) in the embodiment of the present invention × megalobrama amblycephala (♂) offspring.
Embodiment
For the ease of understanding the present invention, hereafter will do to describe more comprehensively, meticulously to the present invention in conjunction with Figure of description and preferred embodiment, but protection scope of the present invention is not limited to following specific embodiment.
Unless otherwise defined, hereinafter used all technical terms are identical with the implication that those skilled in the art understand usually.The object of technical term used herein just in order to describe specific embodiment is not be intended to limit the scope of the invention.
Unless otherwise specified, the various raw material, reagent, instrument and equipment etc. used in the present invention are all bought by market and are obtained or prepare by existing method.
Embodiment:
A kind of method of carp of the present invention and megalobrame amblycephala subfamily distant hydridization, comprise the following steps: first select sexual maturity feature obvious, the female carp that sign is good and male megalobrama amblycephala are cultivated as the special pond of parent population, then in mating season, artificial induced spawning is carried out to described parent population, select the good parent population of spawning effect and carry out artificial dry method insemination, embryo after having inseminated carries out Lotic hatching in water purification hatching apparatus, raise after fry hatching completes, again the fry after raising is detected, screening, measured by fluidic cell DNA content, nephridial tissue ploidy detects and obtains carp and megalobrama amblycephala filial generation, filial generation comprises dliploid Natural Gynogenesis carp, dliploid Natural Gynogenesis mirror carp, the liploid fish of class crucian and Tetraploid fish.
The carp of above-mentioned the present embodiment and the method for megalobrame amblycephala subfamily distant hydridization, specifically comprise the following steps:
The selection of (a) parent population and cultivation: in the present embodiment, the selection of Parent parent population is most important, concrete grammar is before breeding period 3 ~ 4 months, select that sexual maturity feature is obvious, build good, body colour is bright-coloured, physique is healthy and strong, the anosis female carp (see Fig. 1) without wound and male megalobrama amblycephala (see Fig. 2) carry out the cultivation of special pond respectively as the maternal parent population of distant hybridization and male parent parent population, maintenance water quality is good.Particularly rationally to throw something and feed about the breeding period previous moon feed, raise with smart bait, every 2 ~ 3 days stimulation by running water once, to promote parent population gonadal maturation.
(b) artificial induced spawning: in mid-May then to early June, when water temperature stability is more than 20 DEG C, for parent population injection Luteinizing hormone releasing hormone analog (LRH-A) is hastened parturition with the ocyodinic that mixes of human chorionic gonadtropin (HCG), the consumption of LRH-A is the consumption of 10 μ g/kg, HCG is 600IU/kg; First inject maternal parent population, inject male parent parent population again after 4 ~ 5h, the injected dose of male parent parent population reduces by half; By the number ratio of 1:6 ~ 8, maternal parent population, male parent parent population being put into same water surface area after injection is 80m
2in the spawning pond of left and right, then wash by water in pond in time depending on regimen, especially 3 ~ 4h before laying eggs, after injecting certain water yield, stop water filling, allow fish hasten parturition in hydrostatic, until it starts to lay eggs and produce essence smoothly; Adopt pectoral fin base portion without abdominal cavity one, squama place pin injection during injection, to improve the survival rate of parent population.
(c) insemination hatching: after discovery parent population mutually chases in spawning pond, start to draw in the net to salvage, parent population is detected, female carp wet towel good for spawning effect is wrapped up and fixes with canvas stretcher, then carry out artificial dry method insemination with male megalobrama amblycephala, fully stir sperm and ovum with clean chicken feather.Fertilized egg is placed in water purification hatching apparatus carries out Lotic hatching, and water temperature 20 DEG C ~ 21 DEG C, the fry of hatching is raised in pond.
D () raises: after fry all hatches, and prior to cultivating 2 ~ 3 days in incubator, occurring being proceeded to by fry after waist point in the pond of fostering and apply fertilizer in advance raise until fry, to splash after 1 ~ 2 day soya-bean milk, 2 ~ 3 times/day, make every effort to thin, even, can normally ingest until fry grows to.Between the incubation period, the fertility rate and hatchability of statistics embryo.The fertility rate and hatchability of carp and megalobrama amblycephala distant hybridization fish is respectively 85% and 65%.Fry grows up to the offspring chromosome number of somatic of rear nephridial tissue ploidy detection method to carp (♀) × megalobrama amblycephala (♂) and detects.The method that nephridial tissue ploidy detects is: under 18 DEG C ~ 22 DEG C water temperatures, cultural aim fish is after 1 ~ 3 day, PHA is injected 1 ~ 3 time to experiment fish, each dosage is 2 ~ 8 μ g/g body weight, interval time is 12 ~ 24 hours, before dissection is drawn materials 2 ~ 6 hours, injection colchicine, dosage is 2 ~ 4 μ g/g body weight.Take out nephridial tissue, under physiological saline, shred nephridial tissue, 0.075mol/L KCL Hypotonic treatment, at 20 DEG C of temperature hypotonic 40 ~ 60 minutes; With glacial acetic acid: the fixing nephridial tissue of methyl alcohol (1:3) 1 ~ 3 time; Freezing slide drips sheet; Giemsa dye liquor dyeing 45 ~ 60 minutes, thin current rinse the slide back side and remove dye liquor, air-dry rear microscopy, take pictures, and the chromosome map of Four types filial generation is respectively as shown in Fig. 4, Fig. 6, Fig. 8 and Figure 10.
The present embodiment finally obtains dliploid Natural Gynogenesis carp (see Fig. 3), dliploid Natural Gynogenesis mirror carp (see Fig. 5), the liploid fish (see Fig. 7) of class crucian and Tetraploid fish (see Fig. 9) this Four types.
With common red crucian carp for reference, with cells were tested by flow cytometry dliploid Natural Gynogenesis carp, dliploid Natural Gynogenesis mirror carp, erythrocytic DNA average content in the liploid fish of class crucian and this Four types of Tetraploid fish, result is respectively as Figure 12, Figure 13, Figure 14, shown in Figure 15, in analysis chart, data are known, dliploid Natural Gynogenesis carp, dliploid Natural Gynogenesis mirror carp, the liploid fish of class crucian and the DNA average content of Tetraploid fish are respectively 104.28, 93.25, 99.17 with 143.62, dliploid Natural Gynogenesis carp is 1.025 (see Figure 11) with the DNA average content ratio contrasting red crucian carp, without significant difference between the 1:1 theoretical ratio of this ratio and expectation, dliploid Natural Gynogenesis mirror carp is 0.917 with the DNA average content ratio contrasting red crucian carp, without significant difference between the 1:1 theoretical ratio of this ratio and expectation, the liploid fish of class crucian is 0.975 with the DNA average content ratio contrasting red crucian carp, without significant difference between the 1:1 theoretical ratio of this ratio and expectation, Tetraploid fish is 1.412 with the DNA average content ratio contrasting red crucian carp, without significant difference between the 1:1.5 theoretical ratio of this ratio and expectation.Dliploid Natural Gynogenesis carp, dliploid Natural Gynogenesis mirror carp, the liploid fish of class crucian and the DNA average content of Tetraploid fish and be shown in Table 1 with the DNA average content ratio of common red crucian carp:
Table 1: DNA average content and the DNA average content ratio with common red crucian carp in four kinds of filial generations
Fluidic cell DNA content determination method step is: to take a blood sample about 0.2mL from fish tail vein blood vessel with through the wetting disposable syringe of heparin, inject the Eppendorf pipe that 0.8% physiological saline is housed, 1mL nucleus extraction liquid DAPI-A (nuclei extraction solution is added in blood and mixed liquor of normal saline, Germany Partec Gmbh provides), processing time 10 ~ 15min; Sample filters through 20 μm of NFs (German Partec GmbH provides); DNA dyeing liquor (DAPI-B, German Partec GmbH provides) is statically placed in dark place stained specimens about 5 ~ 10min, then goes up machine testing.
The dliploid Natural Gynogenesis carp that the present invention obtains, dliploid Natural Gynogenesis mirror carp, and the liploid fish of similar crucian and this Four types of Tetraploid fish, not only incorporate the multinomial merit of Parent parent population, and, by many for seed selection, be expected to after hybridization generation in cultivate dliploid Natural Gynogenesis carp strain by hereditary and selection, dliploid Natural Gynogenesis mirror carp strain, and the liploid fish strain of similar crucian and Tetraploid fish products system, wherein by continuing seed selection to Tetraploid fish products system, be expected to develop novel autotetraploid fish products system.The present invention has important biological significance in organic evolution and genetics-breeding in fish.
Claims (2)
1. the method for a carp and megalobrame amblycephala subfamily distant hydridization, comprise the following steps: first select sexual maturity feature obvious, the female carp that sign is good and male megalobrama amblycephala are cultivated as the special pond of parent population, then in mating season, artificial induced spawning is carried out to described parent population, select the good parent population of spawning effect and carry out artificial dry method insemination, embryo after having inseminated carries out Lotic hatching in water purification hatching apparatus, raise after fry hatching completes, again the fry after raising is detected, screening, measured by fluidic cell DNA content, nephridial tissue ploidy detects and obtains carp and megalobrama amblycephala filial generation, filial generation comprises dliploid Natural Gynogenesis carp, dliploid Natural Gynogenesis mirror carp, the liploid fish of class crucian and Tetraploid fish.
2. the method for carp according to claim 1 and megalobrame amblycephala subfamily distant hydridization, is characterized in that, described method specifically comprises the following steps:
A the selection of () parent population and cultivation: before breeding period 3 ~ 4 months, select sexual maturity feature is obvious, sign is good female carp and male megalobrama amblycephala and carry out the cultivation of special pond respectively as the maternal parent population of distant hybridization and male parent parent population, keep water quality good; Raise with smart bait in the breeding period previous moon, every 2 ~ 3 days stimulation by running water once;
(b) artificial induced spawning: in mid-May then to early June, when water temperature stability is more than 20 DEG C, for described parent population injection Luteinizing hormone releasing hormone analog is hastened parturition with the ocyodinic that mixes of human chorionic gonadtropin, the consumption of described Luteinizing hormone releasing hormone analog is 6 ~ 10 μ g/kg, and the consumption of human chorionic gonadtropin is 500 ~ 800IU/kg; First inject maternal parent population, inject male parent parent population again after 4 ~ 5h, the injected dose of male parent parent population reduces by half; By the number ratio of 1: 6 ~ 8, maternal parent population, male parent parent population are put into same spawning pond after injection, then wash by water in pond in time depending on regimen, especially 3 ~ 4h before laying eggs, after injecting certain water yield, stop water filling, allow fish hasten parturition in hydrostatic, until it starts to lay eggs and produce essence smoothly;
C () insemination hatching: select the maternal parent population smoothly that lays eggs, fix with canvas stretcher, then measures large male parent parent population carry out artificial dry method insemination with product essence, it is that the water purification hatching apparatus of 20 DEG C ~ 21 DEG C carries out Lotic hatching that fertilized egg is placed in water temperature;
D () raises: after fry all hatches, continue cultivation 2 ~ 3 days in incubator, and occur being proceeded to by fry after waist point in the pond of fostering and apply fertilizer in advance until fry and raise, soya-bean milk of splashing after 1 ~ 2 day, makes every effort to thin, even by 2 ~ 3 times/day; Detect obtain filial generation by fluidic cell DNA content mensuration, nephridial tissue ploidy after fry grows up to.
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| CN105145434A (en) * | 2015-09-12 | 2015-12-16 | 上海海洋大学 | Screening and combining method for megalobrama amblycephala advantage combining |
| CN106135094A (en) * | 2016-06-30 | 2016-11-23 | 湖南文理学院 | The method being efficiently bred as Monopterus albus (Zuiew) male tetraploid pure lines |
| CN106135092A (en) * | 2016-06-30 | 2016-11-23 | 湖南文理学院 | The method improving silver xenocypris fish male tetraploid incubation rate |
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| CN106550909B (en) * | 2016-10-31 | 2019-07-26 | 湖南师范大学 | The method of fancy carp and megalobrame amblycephala subfamily distant hydridization |
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| CN110199918A (en) * | 2019-06-14 | 2019-09-06 | 湖南师范大学 | A kind of method of the method for building up and breeding autotriploid carp of autotetraploid carp strain |
| CN110915727A (en) * | 2019-12-10 | 2020-03-27 | 湖南师范大学 | Method for breeding multiple colorful double-tail goldfish through distant hybridization |
| CN110915727B (en) * | 2019-12-10 | 2021-02-12 | 湖南师范大学 | Method for breeding multiple colorful double-tail goldfish through distant hybridization |
| CN114467808A (en) * | 2021-12-31 | 2022-05-13 | 湖南岳麓山水产育种科技有限公司 | Large-scale preparation method of bighead carp grown from distant hybridization natural gynogenesis |
| CN114698575A (en) * | 2022-05-10 | 2022-07-05 | 湖南文理学院 | Breeding method for variegated carp heterogenous gynogenesis |
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