CN115530098B - Method for breeding cold-resistant fugu obscurus through low-temperature induction - Google Patents
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- 241000980706 Takifugu obscurus Species 0.000 title claims abstract description 75
- 238000009395 breeding Methods 0.000 title claims abstract description 48
- 230000001488 breeding effect Effects 0.000 title claims abstract description 43
- 230000006698 induction Effects 0.000 title claims abstract description 15
- 238000000034 method Methods 0.000 title claims abstract description 15
- 235000013601 eggs Nutrition 0.000 claims abstract description 125
- 241000251468 Actinopterygii Species 0.000 claims abstract description 111
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 39
- 230000000638 stimulation Effects 0.000 claims abstract description 33
- 230000026109 gonad development Effects 0.000 claims abstract description 11
- 238000011161 development Methods 0.000 claims abstract description 7
- 230000018109 developmental process Effects 0.000 claims abstract description 7
- 238000012163 sequencing technique Methods 0.000 claims abstract description 5
- 230000012447 hatching Effects 0.000 claims description 37
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 29
- 230000004720 fertilization Effects 0.000 claims description 17
- 238000002791 soaking Methods 0.000 claims description 16
- 238000001816 cooling Methods 0.000 claims description 15
- 239000012636 effector Substances 0.000 claims description 14
- 238000012216 screening Methods 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 4
- 238000007796 conventional method Methods 0.000 claims description 3
- 230000001568 sexual effect Effects 0.000 claims description 3
- 238000012795 verification Methods 0.000 claims description 3
- 239000002932 luster Substances 0.000 claims description 2
- 239000000284 extract Substances 0.000 claims 1
- 230000000694 effects Effects 0.000 abstract description 7
- 206010064571 Gene mutation Diseases 0.000 abstract description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 230000004936 stimulating effect Effects 0.000 description 2
- 238000009360 aquaculture Methods 0.000 description 1
- 244000144974 aquaculture Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000037406 food intake Effects 0.000 description 1
- 230000002710 gonadal effect Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000001617 migratory effect Effects 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
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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
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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
- A01K61/90—Sorting, grading, counting or marking live aquatic animals, e.g. sex determination
- A01K61/95—Sorting, grading, counting or marking live aquatic animals, e.g. sex determination specially adapted for 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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Abstract
The invention provides a method for breeding cold-resistant fugu obscurus by low-temperature induction, which comprises the steps of firstly carrying out group breeding, and then adopting parent fish and fertilized eggs in gonad development by low-temperature stimulation, so as to improve the probability of gene mutation of egg cell sperms and fertilized eggs in the process of development and division, and facilitate selection of excellent genes and characters; the cold-resistant main effect genes of the bred cold-resistant fugu obscurus are excavated, the gene sequencing is carried out, the gene differences are compared, the cold-resistant main effect genes are found and determined, and the cold-resistant main effect genes are determined, so that the breeding of the cold-resistant fugu obscurus can be carried out at a later stage.
Description
Technical Field
The invention relates to a method for breeding fine varieties of aquaculture varieties, in particular to a breeding method of cold-resistant fugu obscurus.
Background
The Fugu obscurus is the first of Sanxian in Yangtze river, is a fish with long culture, is deeply loved by people at the middle and lower reaches of Yangtze river, is a migratory fish in the sea, and has the most suitable growth temperature of 25-28 ℃, the temperature of 11 months per year is about 18 ℃, and can travel to the sea, and ingestion and death are stopped at 12 ℃. Based on this characteristic of Fugu obscurus, fugu obscurus is difficult to survive in natural conditions in winter in the area downstream of Yangtze river. Therefore, the fugu obscurus cultivated in winter must be heated in a greenhouse, and the temperature is kept at about 18 ℃ to enable the fugu obscurus to pass through winter smoothly.
However, the green house is adopted to heat and cultivate the fugu obscurus, and the fugu obscurus can live through winter, but only maintains the life characteristics of the fugu obscurus, so the fugu obscurus cultivated in the artificial green house grows slowly, and the winter cultivation cost of the fugu obscurus is also seriously increased.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a method for breeding cold-resistant fugu obscurus by low-temperature induction, which can scientifically and effectively breed the fugu obscurus with good cold resistance, thereby improving the yield and weight of the fugu obscurus in winter and further reducing the cost of the fugu obscurus breeding in winter.
The invention provides a method for breeding cold-resistant fugu obscurus by low-temperature induction, which is characterized by comprising the following steps:
s01, breeding parent fish of fugu obscurus, wherein the specific operation is as follows: and selecting target adult fish meeting the requirements of strong physique, good color and luster, good gonad development and maximum specification from the population to serve as the parent fish for reproduction.
S02, carrying out low-temperature induction on the fugu obscurus in the gonad development period, wherein the specific operation is as follows: and (3) performing low-temperature stimulation on the fugu obscurus in the II stage and the III stage of gonad development of the fugu obscurus selected in the S01.
S03, stimulating the fertilized eggs of the fugu obscurus in development at low temperature, wherein the specific operation is as follows: and (3) performing low-temperature stimulation for 30 to 120 minutes after fertilization of the fertilized eggs of the fugu obscurus, wherein the stimulation temperature is 0 to 4 ℃ and the stimulation time is 30 to 120 seconds.
S04, selecting adult fugu obscurus with high growth speed, wherein the specific operation is as follows: the S03 fertilized eggs are hatched into fries, the fries are cultured for about 6 months, a preset culture pond is selected at the beginning of 11 months to the beginning of 12 months, the water temperature of the fish pond is 12 ℃ to 15 ℃, the adult fishes with the largest specification are selected from each culture pond, the adult fishes with the preset growth speed are transferred to the culture pond temporarily cultured in the greenhouse, and each culture pond is numbered.
S05, screening cold-resistant adult fugu obscurus fish, wherein the specific operation is as follows: the temperature of the cultured water is reduced in a gradient way, and the adult fish selected in S04 is divided into small groups with the tolerance temperature of 12 ℃, 10 ℃, 8 ℃, 7 ℃, 6 ℃ and 5 ℃.
S06, excavating cold-resistant main effector genes, wherein the specific operation is as follows: dividing the Fugu obscurus selected in the S05 into 6 groups according to the tolerance temperature, randomly extracting a predetermined number of adult fish from each group, carrying out gene sequencing, comparing and analyzing 2 groups of gene differences of 12 ℃ and 5 ℃ to find out 3-5 cold-resistant main effector genes, comparing and analyzing 4 groups of genes of 10 ℃, 8 ℃, 7 ℃ and 6 ℃ to find out several groups of cold-resistant main effector genes in the 6 groups, namely determining the gene homozygosity of each group, and verifying the cold-resistant main effector genes again.
S07, establishing a cold-resistant parent fish population at 5 ℃, wherein the specific operation is as follows: and selecting adult fish with cold-resistant main effector gene content of more than 3 groups as backup parent fish.
S08, propagation cold-resistant F 1 Instead, cold resistance verification is carried out, and the specific operation is as follows: breeding fries of the backup parent fish to grow adult fish according to a conventional method after sexual maturity, randomly selecting a predetermined number of fish ponds, selecting a predetermined number of adult fish in each pond, screening the breeding method of cold-resistant takifugu obscurus according to the cooling, and verifying F 1 Cold-resistant property of adult fish.
Further, in the step S02, the fugu obscurus in gonadal development stage is induced at a low temperature, and the specific operation is as follows: selecting a preset number of basic parent fishes with male and female parts, controlling the temperature from 18 ℃ to 1-2 ℃ each time, keeping the temperature for 24h, circulating the temperature, cooling to 12 ℃ and keeping the temperature for 48h, eliminating the inferior parent fishes, recovering to above 18 ℃, and continuously culturing.
Further, in the step S03, the fertilized eggs of the fugu obscurus are stimulated at a low temperature, and the specific operation is as follows:
s0301, performing first fertilized egg stimulation, 10 thousands of fertilized eggs, putting the fertilized eggs into water at 0 ℃ for soaking for 30 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond.
S0302, performing second fertilized egg stimulation, namely 10 ten thousand fertilized eggs, putting the fertilized eggs into water at 0 ℃ for 60 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond.
S0303, performing third batch of fertilized egg stimulation, namely 10 ten thousand fertilized eggs, putting the fertilized eggs into water at 0 ℃ for soaking for 120 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond.
S0304, performing fourth fertilized egg stimulation, namely 10 ten thousand fertilized eggs, putting the fertilized eggs into water at 4 ℃ for soaking for 60 seconds after fertilization for 60 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering the fish ponds.
S0305, performing fifth fertilized egg stimulation, namely 10 ten thousand fertilized eggs, putting the fertilized eggs into water at 4 ℃ for soaking for 90 seconds after 90 minutes of fertilization, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond.
S0306, performing sixth batch of fertilized egg stimulation, wherein 10 thousand fertilized eggs are fertilized for 120 minutes, then soaking the fertilized eggs in water at 4 ℃ for 120 seconds, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond.
Further, in the step S05, the specific operation of the gradient cooling is as follows: each time, the temperature is reduced by 1-2 ℃ and each temperature gradient is kept for 24 hours, and the temperature is reduced to 5 ℃ to eliminate the fugu obscurus with poor cold resistance.
The beneficial effects are that: the breeding method for breeding cold-resistant fugu obscurus has the following advantages:
1. the invention adopts parent fish and fertilized eggs in gonad development stimulated at low temperature, so that sperm of egg cells and fertilized eggs are affected by physical, chemical and other factors in the development and division process, the probability of gene mutation is promoted, and the selection of excellent genes and characters is facilitated.
2. The invention discovers and determines the cold-resistant main effector gene by excavating the cold-resistant main effector gene of the bred cold-resistant fugu obscurus and carrying out gene sequencing and comparing the gene differences, thereby facilitating the breeding of the cold-resistant fugu obscurus in the later period.
3. The Fugu obscurus bred by the breeding method has the characteristic of cold resistance, improves the efficiency of Fugu obscurus breeding in winter under natural conditions, and further reduces the cost of Fugu obscurus breeding in winter.
Detailed Description
The invention will be further illustrated with reference to specific examples.
The invention provides a method for breeding cold-resistant fugu obscurus by low-temperature induction, which is characterized by comprising the following steps:
s01, breeding of the parent fish of the fugu obscurus, wherein the parent fish is selected from a group to meet the goals of strong physique, good color, good gonad development and maximum specification.
S02, inducing the fugu obscurus by adopting a gonad development period low-temperature induction method, carrying out low-temperature stimulation on the fugu obscurus selected in S01 in a stage II and a stage III of gonad development of the fugu obscurus, selecting 2000 basic parent fishes of each male and female half, starting to control cooling from 18 ℃, cooling 1 ℃ for the first time, maintaining the temperature for 24 hours, cooling 1.5 ℃ for the second time, maintaining the temperature for 24 hours, cooling 2 ℃ for the third time, maintaining the temperature for 24 hours, cooling 1.5 ℃ for the fourth time, namely cooling to 12 ℃, maintaining the temperature for 48 hours, eliminating the fugu obscurus with poor cold resistance, recovering to more than 18 ℃, and continuing to cultivate.
S03, stimulating the fertilized eggs of the fish seeds in development at a low temperature, wherein the low-temperature stimulation is carried out on the fertilized eggs of the fish seeds for 30-120 min after fertilization, the stimulation temperature is 0-4 ℃, and the stimulation time is 30-120 seconds.
S0301, performing first fertilized egg stimulation, 10 thousands of fertilized eggs, putting the fertilized eggs into water at 0 ℃ for soaking for 30 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond.
S0302, performing second fertilized egg stimulation, namely 10 ten thousand fertilized eggs, putting the fertilized eggs into water at 0 ℃ for 60 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond.
S0303, performing third batch of fertilized egg stimulation, namely 10 ten thousand fertilized eggs, putting the fertilized eggs into water at 0 ℃ for soaking for 120 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond.
S0304, performing fourth fertilized egg stimulation, namely 10 ten thousand fertilized eggs, putting the fertilized eggs into water at 4 ℃ for soaking for 60 seconds after fertilization for 60 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond.
S0305, performing fifth fertilized egg stimulation, namely 10 ten thousand fertilized eggs, putting the fertilized eggs into water at 4 ℃ for soaking for 90 seconds after 90 minutes of fertilization, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond.
S0306, performing sixth batch of fertilized egg stimulation, wherein 10 thousand fertilized eggs are fertilized for 120 minutes, then soaking the fertilized eggs in water at 4 ℃ for 120 seconds, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond.
S04, selecting adult fugu obscurus with high production speed, hatching the fertilized eggs in the S03 into fish fries, culturing the fish fries for about 6 months, selecting 18 culture ponds in the S03 at the beginning of 11 months to the beginning of 12 months when the water temperature of the fish ponds is 12-15 ℃, selecting 1000 fish with the largest production speed in each culture pond, transferring the culture ponds temporarily cultured in a greenhouse, wherein the area of each culture pond is 50-60 square meters, and numbering each culture pond.
S05, cooling, screening cold-resistant adult fish of takifugu obscurus, adjusting the water temperature of cultivation to 12 ℃, keeping for 24 hours, selecting adult fish resistant to 12 ℃, and numbering the adult fish; continuously adjusting the water temperature of the culture to 10 ℃, keeping for 24 hours, selecting adult fish resistant to 10 ℃, and numbering the adult fish; continuously adjusting the water temperature of the culture to 8 ℃, keeping for 24 hours, selecting adult fish resistant to 8 ℃, and numbering the adult fish; continuously adjusting the water temperature of the culture to 7 ℃, keeping for 24 hours, selecting adult fish resistant to 7 ℃, and numbering the adult fish; continuously adjusting the water temperature of the culture to 6 ℃, keeping for 24 hours, selecting adult fish resistant to the temperature of 6 ℃ and numbering the adult fish; continuously adjusting the water temperature of the culture to 5 ℃, keeping for 24 hours, selecting adult fish with the temperature of 5 ℃ resistance, and numbering the adult fish.
S06, excavating cold-resistant main effect genes, randomly extracting 6 groups of groups selected in the S05, selecting 50 fish from each group of groups, carrying out gene sequencing, comparing and analyzing 2 groups of gene differences of 12 ℃ and 5 ℃, finding and determining 3-5 cold-resistant main effect genes, comparing and analyzing 4 groups of genes of 10 ℃, 8 ℃, 7 ℃ and 6 ℃ to find and determine the gene homozygosity in each group of the cold-resistant main effect genes, and determining the cold-resistant main effect genes again.
S07, establishing a cold-resistant parent fish population at 5 ℃, and selecting adult fish with the cold-resistant main effector gene content of more than 3 groups as backup parent fish.
S08, propagation cold-resistant F 1 Performing cold resistance verification, breeding fries of the reserve parent fish to grow into adult fish according to a conventional method after sexual maturity, randomly selecting a predetermined number of fish ponds, selecting a predetermined number of adult fish in each pond, screening the breeding method of the cold-resistant adult fish according to the cooling, and verifying F 1 Cold resistance of adult fish.
The adult fish of the selected fugu obscurus spawns in 4 months to 5 months, fish fries are cultivated to 5 ℃ in 12 months, meanwhile, sampling measurement analysis is carried out on the selected fugu obscurus and the non-selected fugu obscurus, wherein the average weight of the selected fugu obscurus is 249.6+/-50.12 g, and the adult fish still has full vitality in the environment of 5 ℃; the average weight of the unoccupied Fugu obscurus is 110.3+/-28.93 g, and most of the Fugu obscurus stops feeding at 18 ℃ at the beginning of 11 months and dies gradually, and all of the Fugu obscurus dies at 5 ℃. The result shows that the bred fugu obscurus has obvious cold resistance.
The described embodiments of the present invention are intended to be only a few, but not all embodiments of the present invention, and all other embodiments that may be made by one of ordinary skill in the art without inventive faculty are intended to be within the scope of the present invention.
Claims (4)
1. A method for breeding cold-resistant Fugu obscurus by low-temperature induction is characterized in that: the method comprises the following specific steps:
s01, breeding parent fish of fugu obscurus, wherein the specific operation is to select target adult fish meeting the requirements of strong physique, good color and luster, good gonad development and maximum specification from a group to serve as the parent fish for breeding;
s02, carrying out low-temperature induction on the fugu obscurus in the gonad development stage, wherein the specific operation is that the fugu obscurus is subjected to low-temperature stimulation induction in the stages II and III of the gonad development of the fugu obscurus selected in the S01;
s03, low-temperature stimulation of fertilized eggs of the fugu obscurus in development, wherein the specific operation is that the fertilized eggs of the fugu obscurus are subjected to low-temperature stimulation for 30 to 120 minutes at the stimulation temperature of 0 to 4 ℃ and the stimulation time of 30 to 120 seconds;
s04, selecting adult fugu obscurus with high growth speed, wherein the specific operation is as follows: incubating fertilized eggs into fish fries in the step S03, culturing the fish fries for about 6 months, selecting a preset culture pond when the water temperature of the fish pond is 12-15 ℃ at the beginning of 11 months-12 months, selecting a preset number of fish with the largest specification in each culture pond, transferring the culture ponds temporarily cultured in a greenhouse, and numbering each culture pond;
s05, screening cold-resistant adult fugu obscurus fish, wherein the specific operation is as follows: gradient cooling is carried out on the cultivation water temperature, and the adult fugu obscurus selected in S04 is divided into small groups with the temperature resistance of 12 ℃, 10 ℃, 8 ℃, 7 ℃, 6 ℃ and 5 ℃ according to the tolerance temperature;
s06, excavating cold-resistant main effector genes, wherein the specific operation is as follows: the Fugu obscurus adult fish selected in the step S05 is divided into 6 groups of groups, each group randomly extracts a preset number of Fugu obscurus, and gene sequencing is carried out; then comparing and analyzing the difference between the 2 groups of genes at 12 ℃ and 5 ℃ to find and determine 3-5 cold-resistant main effector genes; after the cold-resistant main effector genes are determined, continuing to control and analyze 4 groups of genes at 10 ℃, 8 ℃, 7 ℃ and 6 ℃ to determine the gene homozygosity in each group of the cold-resistant main effector genes, and verifying the cold-resistant main effector genes again;
s07, establishing a cold-resistant parent fish population at 5 ℃, wherein the specific operation is as follows: selecting adult fugu obscurus with cold-resistant main effector gene content of more than 3 groups as backup parent fish;
s08, propagation cold-resistant F 1 Carrying out cold resistance verification on the generation; the specific operation is as follows: breeding fries of the reserve parent fish to grow adult fish according to a conventional method after sexual maturity, randomly selecting a predetermined number of fish ponds, selecting a predetermined number of adult fish in each pond, and gradient cooling and screening cold-resistant adult fish according to the step S05Verify F 1 Cold-resistant property of adult fish.
2. The method for breeding cold-resistant fugu obscurus by low-temperature induction according to claim 1, wherein the specific operation of low-temperature induction of fugu obscurus in the step S02 is as follows: selecting a preset number of basic parent fishes with male and female parts, controlling the temperature from 18 ℃ to 1-2 ℃ each time, keeping the temperature for 24h, circulating the temperature, cooling to 12 ℃ and keeping the temperature for 48h, eliminating the inferior parent fishes, recovering to above 18 ℃, and continuously culturing.
3. The method for breeding cold-resistant fugu obscurus by low-temperature induction according to claim 1, wherein in the step S03, the specific operation of low-temperature stimulation of the fertilized eggs of the fugu obscurus in development is as follows:
s0301, performing first fertilized egg stimulation, namely 10 thousands of fertilized eggs, putting the fertilized eggs into water at 0 ℃ for soaking for 30 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond;
s0302, performing second fertilized egg stimulation, namely 10 thousands of fertilized eggs, putting the fertilized eggs into water at 0 ℃ for soaking for 60 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond;
s0303, performing third batch of fertilized egg stimulation, namely 10 thousands of fertilized eggs, putting the fertilized eggs into water at 0 ℃ for soaking for 120 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond;
s0304, performing fourth fertilized egg stimulation, namely 10 thousands of fertilized eggs, putting the fertilized eggs into water at 4 ℃ for soaking for 60 seconds after fertilization for 60 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond;
s0305, performing fifth batch of fertilized egg stimulation, namely 10 ten thousand fertilized eggs, putting the fertilized eggs into water at 4 ℃ for soaking for 90 seconds after 90 minutes of fertilization, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond;
s0306, performing sixth batch of fertilized egg stimulation, wherein 10 thousand fertilized eggs are fertilized for 120 minutes, then soaking the fertilized eggs in water at 4 ℃ for 120 seconds, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs according to the normal temperature, breeding the fry in 3 fish ponds, and numbering each fish pond.
4. The method for breeding cold-resistant Fugu obscurus by low-temperature induction according to claim 1, wherein in the step S05, the specific operation of gradient cooling is as follows: each time, the temperature is reduced by 1-2 ℃ and each temperature gradient is kept for 24 hours, and the temperature is reduced to 5 ℃ to eliminate the fugu obscurus with poor cold resistance.
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