CN109362616A - A kind of breeding method of triploid carp - Google Patents
A kind of breeding method of triploid carp Download PDFInfo
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- CN109362616A CN109362616A CN201811541945.9A CN201811541945A CN109362616A CN 109362616 A CN109362616 A CN 109362616A CN 201811541945 A CN201811541945 A CN 201811541945A CN 109362616 A CN109362616 A CN 109362616A
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- 208000026487 Triploidy Diseases 0.000 title claims abstract description 61
- 241000252233 Cyprinus carpio Species 0.000 title claims abstract description 51
- 238000009395 breeding Methods 0.000 title claims abstract description 19
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 62
- 230000002706 hydrostatic effect Effects 0.000 claims abstract description 50
- 235000013601 eggs Nutrition 0.000 claims abstract description 47
- 238000012545 processing Methods 0.000 claims abstract description 31
- 238000000034 method Methods 0.000 claims abstract description 27
- 102000002322 Egg Proteins Human genes 0.000 claims abstract description 25
- 108010000912 Egg Proteins Proteins 0.000 claims abstract description 25
- 210000004681 ovum Anatomy 0.000 claims abstract description 25
- 230000009027 insemination Effects 0.000 claims abstract description 17
- 230000035800 maturation Effects 0.000 claims abstract description 9
- 238000002156 mixing Methods 0.000 claims abstract description 9
- IAKHMKGGTNLKSZ-INIZCTEOSA-N (S)-colchicine Chemical compound C1([C@@H](NC(C)=O)CC2)=CC(=O)C(OC)=CC=C1C1=C2C=C(OC)C(OC)=C1OC IAKHMKGGTNLKSZ-INIZCTEOSA-N 0.000 claims description 40
- 229960001338 colchicine Drugs 0.000 claims description 20
- 241000251468 Actinopterygii Species 0.000 abstract description 28
- 230000006698 induction Effects 0.000 abstract description 8
- 230000001488 breeding effect Effects 0.000 abstract description 2
- 238000006243 chemical reaction Methods 0.000 abstract description 2
- 241000790704 Cyprinus carpio 'Songpu mirror carp' Species 0.000 description 19
- 210000003743 erythrocyte Anatomy 0.000 description 18
- 239000000243 solution Substances 0.000 description 16
- 210000000349 chromosome Anatomy 0.000 description 8
- 238000003825 pressing Methods 0.000 description 8
- 108020004414 DNA Proteins 0.000 description 7
- 230000026109 gonad development Effects 0.000 description 6
- 210000004940 nucleus Anatomy 0.000 description 6
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 238000004043 dyeing Methods 0.000 description 5
- 210000004907 gland Anatomy 0.000 description 5
- 238000002386 leaching Methods 0.000 description 5
- 230000001568 sexual effect Effects 0.000 description 5
- 230000009466 transformation Effects 0.000 description 5
- 239000002504 physiological saline solution Substances 0.000 description 4
- 210000001550 testis Anatomy 0.000 description 4
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- BVIAOQMSVZHOJM-UHFFFAOYSA-N N(6),N(6)-dimethyladenine Chemical compound CN(C)C1=NC=NC2=C1N=CN2 BVIAOQMSVZHOJM-UHFFFAOYSA-N 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 2
- 244000144974 aquaculture Species 0.000 description 2
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- 229960000549 4-dimethylaminophenol Drugs 0.000 description 1
- VHYFNPMBLIVWCW-UHFFFAOYSA-N 4-dimethylaminopyridine Substances CN(C)C1=CC=NC=C1 VHYFNPMBLIVWCW-UHFFFAOYSA-N 0.000 description 1
- 241000252210 Cyprinidae Species 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
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- OOYGSFOGFJDDHP-KMCOLRRFSA-N kanamycin A sulfate Chemical group OS(O)(=O)=O.O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CN)O[C@@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](N)[C@H](O)[C@@H](CO)O2)O)[C@H](N)C[C@@H]1N OOYGSFOGFJDDHP-KMCOLRRFSA-N 0.000 description 1
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01K—ANIMAL HUSBANDRY; AVICULTURE; APICULTURE; PISCICULTURE; FISHING; REARING OR BREEDING ANIMALS, NOT OTHERWISE PROVIDED FOR; NEW BREEDS OF ANIMALS
- A01K61/00—Culture of aquatic animals
- A01K61/10—Culture of aquatic animals of fish
-
- 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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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
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- Farming Of Fish And Shellfish (AREA)
Abstract
The invention discloses a kind of breeding methods of triploid carp, are related to carp breeding technical field, the technical scheme comprises the following steps: taking the ovum and sperm of carp maturation, are sprinkling upon on sieve after mixing, are immersed in the water completion insemination, obtain fertilized eggs；It immerses in cold water and handles after progress hydrostatic pressure processing；It is transferred in water and hatches, obtain triploid carp.Method provided by the invention is convenient for execute-in-place, safe and reliable, simple and easy, triploid high conversion rate, has significant application value in cyprinid fish triploid induction and large-scale cultivation.
Description
Technical field
The present invention relates to carp breeding technical fields, and in particular to a kind of breeding method of triploid carp.
Background technique
Nature overwhelming majority fish are raised up seed as existing for diploid form, and in a manner of amphimictic,
The sexal maturity stage of diploid fish, gonad development consume big energy, and due to gonad development stage and spawning season fish
Meat quality declines and extends Time To Market, meanwhile, gonad development stage individual growth is stagnated and the death rate increases, and individual body
Type is very limited.Natural diploid fish need artificial induction that could generate triploid.The side of artificial induction's triploid fish
Method mainly has the physical methods such as cold shock, heat shock, hydrostatic pressing, using chemical substance (such as 6- dimethylaminopurine (6-
DMAP), colchicine etc.) processing chemical method.Inhibit fertilization with methods such as temperature shock, hydrostatic pressing, drug-treateds at present
Ovum second polar body outlet makes genome tripling to produce triploid fish and at home and abroad also have many reports, but it is most of still
So in exploration and experimental stage, still there is a certain distance from practical application, many results are optimal still without finding out completely
Phenomena such as induction parameters, fertilized eggs necrosis rate is high, emergence rate is low, juvenile fish abnormal rate is high and tripling ratio is relatively low, affect three
The process of times body industrialization.
It is complete if advanced optimizing the treatment conditions of triploid induction in terms of the current situation of current aquaculture industry
The abductive approach and techniqueflow of kind triploid, it will there are more triploid fish to apply in culture fishery, promote fish
The sound development of class aquaculture.
Summary of the invention
To solve the above problems, the present invention provides a kind of breeding method of triploid carp, technical solution includes following step
It is rapid:
1) ovum and sperm for taking carp maturation, are sprinkling upon on sieve after mixing, are immersed in the water completion insemination, are fertilized
Ovum；
2) fertilized eggs for obtaining step 1) are put into hydrostatic pressure device storehouse, carry out hydrostatic pressure processing；
3) step 2) hydrostatic pressure treated fertilized eggs are taken out, immerses in cold water and handles；
4) fertilized eggs for obtaining step 3), which are transferred in water, hatches.
The preferred mirror carp of the step 1) carp, more preferable Songpu mirror carp.
The volume ratio of the step 1) sperm and ovum is 500 μ L:(50-100) mL.
Water temperature in step 1) is 23-25 DEG C.
After the completion of the step 1) Process of Insemination, fertilized eggs continue to develop 3-4.5min in water.
There are the colchicine solution of 500 μ g/ml, the leaching of fertilized eggs obtained by step 1) in step 2) hydrostatic pressure device storehouse
Enter in colchicine solution.
Step 2) the hydrostatic pressure processing, pressure 600kg/cm2, the processing time is 3min.
In step 2) the hydrostatic pressure treatment process, colchicine solution temperature is 23-25 DEG C.
Step 3) the cold water temperature is 0-2 DEG C, and the processing time is 10-15min.
Water temperature in step 4) is 23-25 DEG C.
Beneficial effect
Method provided by the invention utilizes method provided by the invention training convenient for execute-in-place, safe and reliable, simple and easy
Triploid carp is educated, being examined three times transformation rate is 91%, the triploid carp cultivated, compared with common diploid,
Erythrocyte volume ratio be 1.63:1, erythrocyte nucleus volume ratio be 1.56:1, triploid fish erythrocyte nucleus major diameter, red blood cell area,
Compared with diploid, difference is extremely significant for long term voyage and Assessment of Nuclear Volume；Triploid carp sexual gland agensis, spermary sections observation are presented
Large area vacuolation；The comparison of 3 age fishes, triploid carp and diploid carp weight ratio are 1.7 ± 0.21.It is cultivated using this method
Triploid carp, there is no the sexal maturity stage gonad developments of diploid fish to consume big energy, due to gonad development rank
Section and spawning season flesh of fish quality decline and extend Time To Market, gonad development stage individual growth is stagnated and the death rate increases,
The problems such as, there is significant application value in cyprinid fish triploid induction and large-scale cultivation.
Detailed description of the invention
Fig. 1 diploid (3 age group) compared with triploid carp size；
3 age of Fig. 2 diploid and triploid sexual gland contrast difference；Wherein 2n indicates diploid, and 3n indicates triploid；
Fig. 33 age diploid carp testis histotomy, 40 times of photos of optical microscopy；
Fig. 43 age triploid carp testis histotomy, 40 times of photos of optical microscopy；
Fig. 5 diploid Songpu mirror carp red blood cell (HE dyeing)；
Fig. 6 triploid Songpu mirror carp red blood cell (HE dyeing)；
Fig. 7 diploid Songpu mirror carp division phases；
Fig. 8 triploid Songpu mirror carp division phases；
Fig. 9 diploid Songpu mirror carp DNA content；
Figure 10 triploid Songpu mirror carp DNA content.
Specific embodiment
Embodiment 1
A kind of breeding method of triploid carp, comprising the following steps:
1) ovum and sperm for taking carp maturation, are sprinkling upon on sieve after mixing, are immersed in the water completion insemination, are fertilized
Ovum；
2) sieve with fertilized eggs that step 1) obtains is folded rapidly and is put into hydrostatic pressure device storehouse, carry out hydrostatic pressing
Power processing；
3) step 2) hydrostatic pressure treated fertilized eggs are taken out, immerses in cold water and handles；
4) fertilized eggs for obtaining step 3), which are transferred in water, hatches.
Step 1) the carp is Songpu mirror carp.
The volume ratio of the step 1) sperm and ovum is 500 μ L:100mL.
Water temperature in step 1) is 23 DEG C.
After the completion of the step 1) Process of Insemination, fertilized eggs continue to develop 3min in water.
There are the colchicine solution of 500 μ g/ml, the leaching of fertilized eggs obtained by step 1) in step 2) hydrostatic pressure device storehouse
Enter in colchicine solution.
Step 2) the hydrostatic pressure processing, pressure 600kg/cm2, the processing time is 3min.
In step 2) the hydrostatic pressure treatment process, colchicine solution temperature is 23 DEG C.
Step 3) the cold water temperature is 0 DEG C, and the processing time is 10min.
Water temperature in step 4) is 23 DEG C.
Step 1) -3) fertilized eggs sieve carrying under operated.
Embodiment 2
A kind of breeding method of triploid carp, comprising the following steps:
1) ovum and sperm for taking carp maturation, are sprinkling upon on sieve after mixing, are immersed in the water completion insemination, are fertilized
Ovum；
2) sieve with fertilized eggs that step 1) obtains is folded rapidly and is put into hydrostatic pressure device storehouse, carry out hydrostatic pressing
Power processing；
3) step 2) hydrostatic pressure treated fertilized eggs are taken out, immerses in cold water and handles；
4) fertilized eggs for obtaining step 3), which are transferred in water, hatches.
Step 1) the carp is Songpu mirror carp.
The volume ratio of the step 1) sperm and ovum is 500 μ L:50mL.
Water temperature in step 1) is 25 DEG C.
After the completion of the step 1) Process of Insemination, fertilized eggs continue to develop 4.5min in water.
There are the colchicine solution of 500 μ g/ml, the leaching of fertilized eggs obtained by step 1) in step 2) hydrostatic pressure device storehouse
Enter in colchicine solution.
Step 2) the hydrostatic pressure processing, pressure 600kg/cm2, the processing time is 3min.
In step 2) the hydrostatic pressure treatment process, colchicine solution temperature is 25 DEG C.
Step 3) the cold water temperature is 2 DEG C, and the processing time is 15min.
Water temperature in step 4) is 25 DEG C.
Step 1) -3) fertilized eggs sieve carrying under operated.
Embodiment 3
A kind of breeding method of triploid carp, comprising the following steps:
1) ovum and sperm for taking carp maturation, are sprinkling upon on sieve after mixing, are immersed in the water completion insemination, are fertilized
Ovum；
2) sieve with fertilized eggs that step 1) obtains is folded rapidly and is put into hydrostatic pressure device storehouse, carry out hydrostatic pressing
Power processing；
3) step 2) hydrostatic pressure treated fertilized eggs are taken out, immerses in cold water and handles；
4) fertilized eggs for obtaining step 3), which are transferred in water, hatches.
Step 1) the carp is Songpu mirror carp.
The volume ratio of the step 1) sperm and ovum is 500 μ L:80mL.
Water temperature in step 1) is 24 DEG C.
After the completion of the step 1) Process of Insemination, fertilized eggs continue to develop 4min in water.
There are the colchicine solution of 500 μ g/ml, the leaching of fertilized eggs obtained by step 1) in step 2) hydrostatic pressure device storehouse
Enter in colchicine solution.
Step 2) the hydrostatic pressure processing, pressure 600kg/cm2, the processing time is 3min.
In step 2) the hydrostatic pressure treatment process, colchicine solution temperature is 24 DEG C.
Step 3) the cold water temperature is 1 DEG C, and the processing time is 13min.
Water temperature in step 4) is 24 DEG C.
Step 1) -3) fertilized eggs sieve carrying under operated.
Comparative example
Experimental group 1
1) ovum and sperm for taking carp maturation, are sprinkling upon on sieve after mixing, are immersed in the water completion insemination, are fertilized
Ovum；
2) sieve with fertilized eggs that step 1) obtains is folded rapidly and is put into hydrostatic pressure device storehouse, carry out hydrostatic pressing
Power processing；
3) step 2) hydrostatic pressure treated fertilized eggs are taken out, is transferred in water and hatches；
Step 1) the carp is Songpu mirror carp.
The volume ratio of the step 1) sperm and ovum is 500 μ L:80mL.
Water temperature in step 1) is 24 DEG C.
After the completion of the step 1) Process of Insemination, fertilized eggs continue to develop 4min in water.
There is water in step 2) hydrostatic pressure device storehouse, fertilized eggs obtained by step 1) are immersed in the water.
Step 2) the hydrostatic pressure processing, pressure 600kg/cm2, the processing time is 3min.
Water temperature in step 3) is 24 DEG C.
Step 1) -2) fertilized eggs sieve carrying under operated.
Experimental group 2
1) ovum and sperm for taking carp maturation, are sprinkling upon on sieve after mixing, are immersed in the water completion insemination, are fertilized
Ovum；
2) sieve with fertilized eggs that step 1) obtains is folded rapidly and is put into hydrostatic pressure device storehouse, carry out hydrostatic pressing
Power processing；
3) step 2) hydrostatic pressure treated fertilized eggs are taken out, immerses in cold water and handles；
4) fertilized eggs for obtaining step 3), which are transferred in water, hatches.
Step 1) the carp is Songpu mirror carp.
The volume ratio of the step 1) sperm and ovum is 500 μ L:80mL.
Water temperature in step 1) is 24 DEG C.
After the completion of the step 1) Process of Insemination, fertilized eggs continue to develop 4min in water.
There is water in step 2) hydrostatic pressure device storehouse, fertilized eggs obtained by step 1) are immersed in the water.
Step 2) the hydrostatic pressure processing, pressure 600kg/cm2, the processing time is 3min.
In step 2) the hydrostatic pressure treatment process, coolant-temperature gage is 24 DEG C.
Step 3) the cold water temperature is 1 DEG C, and the processing time is 13min.
Water temperature in step 4) is 24 DEG C.
Step 1) -3) fertilized eggs sieve carrying under operated.
Experimental group 3
1) ovum and sperm for taking carp maturation, are sprinkling upon on sieve after mixing, are immersed in the water completion insemination, are fertilized
Ovum；
2) sieve with fertilized eggs that step 1) obtains is folded rapidly and is put into hydrostatic pressure device storehouse, carry out hydrostatic pressing
Power processing；
3) step 2) hydrostatic pressure treated fertilized eggs are taken out, is transferred in water and hatches；
Step 1) the carp is Songpu mirror carp.
The volume ratio of the step 1) sperm and ovum is 500 μ L:80mL.
Water temperature in step 1) is 24 DEG C.
After the completion of the step 1) Process of Insemination, fertilized eggs continue to develop 4min in water.
There are the colchicine solution of 500 μ g/ml, the leaching of fertilized eggs obtained by step 1) in step 2) hydrostatic pressure device storehouse
Enter in colchicine solution.
Step 2) the hydrostatic pressure processing, pressure 600kg/cm2, the processing time is 3min.
In step 2) the hydrostatic pressure treatment process, colchicine solution temperature is 24 DEG C.
Water temperature in step 3 is 24 DEG C.
Step 1) -2) fertilized eggs sieve carrying under operated.
Result verification:
Triploid identification method
1) erythrocyte volume identification method
From test fish tail vein blood, blood film, HE dyeing are prepared, neutral optical natural gum mounting measures red under oil mirror
Cell and the long and short diameter of core, 10 red blood cells of every blood film random measurement and core size, according to formula ab2/ 1.91 wherein a be
Major diameter, b is the volume that minor axis calculates red blood cell and core, and determines identification ploidy by two, triploid red blood cell major diameter range
Shi Suoyong major diameter standard.
1) chromosome karyotype analysis
Every tail injects PHA (calf serum dissolution) according to 5 μ l/g pectoral fins, 12~colchicine presses 1.5 μ g/g weight afterwards for 24 hours
Ratio is injected into fish body, is docked after two hours, is cut gill bloodletting, and head-kidney is taken to clean 3~5 times in 0.75% physiological saline,
1000r/min is centrifuged 5~7min after grinding, takes supernatant, and 0.5%KCl room temperature handles 40min, cell is collected by centrifugation, solid with Kano
Determine liquid to fix 3 times, no less than 15min, ice slide drip piece every time, and standing is dried, and Giemsa dyes 20min, after rinsing with ruinning water
It dries, microscopically observation.
3) DNA content detects
The complete 900 μ l physiological saline (0.75%) of fish blood fish of 300~400 μ l is sufficiently mixed, by mixed liquor by fish venous blood sampling
It is slowly added dropwise on 6ml human lymphocyte separating liquid liquid level, 2500~3000rpm of room temperature is centrifuged 20min, supernatant is abandoned, with life
It manages and 1ml physiological saline (0.75%) resuspension is added after salt water (0.75%) rinses 2~3 times, take 100 μ l cell suspensions pre- in 6ml
Cold Kano fixer is fixed (glacial acetic acid: methanol=1:3), 4 DEG C of 2h, and supernatant is abandoned in 800rpm, 5min centrifugation, and 6ml physiology is added
Salt water (0.75%), 600rpm, 3min rinsing, is repeated 3 times, and final 1ml physiological saline is resuspended, and PI is protected from light dyeing, 4 DEG C of dyeing
30min, upper machine, chicken blood are compareed with standard diploid after the filtering of 400 mesh.
(in following result, triploid is the triploid carp obtained of embodiment 3 to experimental result, and diploid is same item
The common diploid carp raised under part)
1, Apparent character is observed
Fig. 1 is diploid compared with triploid carp (embodiment 3) size, in 3 ages and 2 age groups, 3 times of bodies and 2 times of body carps,
There is significant difference in weight and metric characters, in 3 age groups, weight 3n/2n=1.7 ± 0.21.
2, sexual gland phenotype
Fig. 2 is 3 age diploids and triploid (embodiment 3) sexual gland contrast difference, it can be seen that 3 times of body sexual gland abortions；It is logical
Cross spermary slice comparison (Fig. 3,4), it is seen that triploid carp testis slice is sliced compared to diploid carp testis, and large area is presented
Vacuolation.
3, the observation of erythrocyte nucleus size and statistics
Table 1 two, triploid Erythrocyte measure value and ratio
**It indicates to be in extremely significant difference
The size of the red blood cell and nucleus of liploid fish and triploid fish (embodiment 3) and significant difference between the two
Property inspection result is shown in Table 1.Triploid fish items numerical value is all higher than liploid fish it can be seen from table, shows pole therebetween
Significant difference.The volume ratio of liploid fish and triploid fish is 1:1.63, and erythrocyte nucleus volume ratio is 1:1.56, and triploid fish is red
Compared with diploid, difference is extremely significant (P ﹤ 0.001) for nucleus major diameter, red blood cell area, long term voyage and Assessment of Nuclear Volume.
4, chromosome detects
Diploid Songpu mirror carp chromosome number is 2n=100, and triploid Songpu mirror carp (embodiment 3) chromosome number is
3n=150, is shown in Fig. 7, Fig. 8, and diploid Songpu mirror carp and triploid Songpu mirror carp chromosome number are consistent with theory expectation.
5, DNA content detects
Songpu mirror carp 2n and Songpu mirror carp 3n (embodiment 3) DNA relative amount are shown in Fig. 9, Figure 10, triploid sample chromosomes
Number and DNA content are about 1:1.58 in the ratio of standard diploid, are consistent with erythrocyte size measurement result.Chromosome number
Mesh and DNA content meet proportionate relationship, are consistent with erythrocyte size measurement result.
The nuclear DNA content ratio of triploid and diploid is about 1:1.58, with red blood cell diameter, volume and chromosome
Number is positively correlated.
6, the tripling ratio (comparative example) of different methods for inducing
By the method for above-mentioned identification triploid, each experimental group identifies 100 tails, determines each experimental group tripling ratio.See
Table 2.
2 different experiments group induced triploid ratio of table
1 three times transformation rate of experimental group is 53%, and 2 three times transformation rate of experimental group is 67%, the conversion of 3 triploid of experimental group
Rate is 62%, and 3 three times transformation rate of the embodiment of the present invention is 91%, in addition the embodiment of the present invention 1,2 three times transformation rates difference
For 83% and 87%, illustrate that the present invention greatly improves triploid induction success rate, has and be convenient for execute-in-place, safety can
The advantages that leaning on, is simple and easy has significant application value in cyprinid fish triploid induction and large-scale cultivation.
Claims (9)
1. a kind of breeding method of triploid carp, it is characterised in that: the following steps are included:
1) ovum and sperm for taking carp maturation, are sprinkling upon on sieve after mixing, are immersed in the water completion insemination, obtain fertilized eggs；
2) fertilized eggs for obtaining step 1) are put into hydrostatic pressure device storehouse, carry out hydrostatic pressure processing；
3) step 2) hydrostatic pressure treated fertilized eggs are taken out, immerses in cold water and handles；
4) fertilized eggs for obtaining step 3), which are transferred in water, hatches.
2. the breeding method of triploid carp according to claim 1, it is characterised in that: the step 1) sperm and ovum
Volume ratio is 500 μ L:(50-100) mL.
3. the breeding method of triploid carp according to claim 1, it is characterised in that: the water temperature in step 1) is 23-
25℃。
4. the breeding method of triploid carp according to claim 1, it is characterised in that: the step 1) Process of Insemination is complete
Cheng Hou, fertilized eggs continue to develop 3-4.5min in water.
5. the breeding method of triploid carp according to claim 1, it is characterised in that: step 2) the hydrostatic pressure device
There is the colchicine solution of 500 μ g/ml in storehouse, fertilized eggs obtained by step 1) immerse in colchicine solution.
6. the breeding method of triploid carp according to claim 1, it is characterised in that: at the step 2) hydrostatic pressure
Reason, pressure 600kg/cm2, the processing time is 3min.
7. the breeding method of triploid carp according to claim 1, it is characterised in that: at the step 2) hydrostatic pressure
During reason, colchicine solution temperature is 23-25 DEG C.
8. the breeding method of triploid carp according to claim 1, it is characterised in that: the step 3) cold water temperature is
0-2 DEG C, the processing time is 10-15min.
9. the breeding method of triploid carp according to claim 1, it is characterised in that: the water temperature in step 4) is 23-
25℃。
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| CN115067277A (en) * | 2022-06-09 | 2022-09-20 | 东北农业大学 | Method for cultivating triploid culter alburnus |
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| 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 |
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