CN115530098A - Method for breeding cold-resistant fugu obscurus through low-temperature induction - Google Patents
Method for breeding cold-resistant fugu obscurus through low-temperature induction Download PDFInfo
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- 241000980706 Takifugu obscurus Species 0.000 title claims abstract description 80
- 238000009395 breeding Methods 0.000 title claims abstract description 50
- 230000001488 breeding effect Effects 0.000 title claims abstract description 44
- 238000000034 method Methods 0.000 title claims abstract description 22
- 230000006698 induction Effects 0.000 title claims abstract description 13
- 241000251468 Actinopterygii Species 0.000 claims abstract description 114
- 235000013601 eggs Nutrition 0.000 claims abstract description 97
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 38
- 230000004936 stimulating effect Effects 0.000 claims abstract description 23
- 230000000694 effects Effects 0.000 claims abstract description 20
- 238000011161 development Methods 0.000 claims abstract description 10
- 230000018109 developmental process Effects 0.000 claims abstract description 10
- 230000026109 gonad development Effects 0.000 claims abstract description 8
- 238000012163 sequencing technique Methods 0.000 claims abstract description 5
- 230000012447 hatching Effects 0.000 claims description 32
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 32
- 230000004720 fertilization Effects 0.000 claims description 18
- 238000002791 soaking Methods 0.000 claims description 18
- 238000001816 cooling Methods 0.000 claims description 16
- 230000000638 stimulation Effects 0.000 claims description 15
- 238000012216 screening Methods 0.000 claims description 8
- 230000002710 gonadal effect Effects 0.000 claims description 5
- 230000001939 inductive effect Effects 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 4
- 238000007796 conventional method Methods 0.000 claims description 3
- 238000011534 incubation Methods 0.000 claims description 3
- 230000001902 propagating effect Effects 0.000 claims description 3
- 238000012795 verification Methods 0.000 claims description 2
- 206010064571 Gene mutation Diseases 0.000 abstract description 2
- 210000004602 germ cell Anatomy 0.000 description 2
- 238000010438 heat treatment Methods 0.000 description 2
- 230000001976 improved effect Effects 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 244000144974 aquaculture Species 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 235000013305 food Nutrition 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 performing group breeding, and stimulating parent fish and fertilized eggs in gonad development at low temperature to improve the probability of gene mutation of egg cell sperms and fertilized eggs in the development and division process, thereby facilitating selection of excellent genes and characters; and (3) excavating cold-resistant major effect genes of the bred cold-resistant fugu obscurus, performing gene sequencing, comparing gene differences, finding and determining the cold-resistant major effect genes, and facilitating breeding of the cold-resistant fugu obscurus at the later stage.
Description
Technical Field
The invention relates to a method for breeding improved varieties of aquaculture species, in particular to a breeding method of cold-resistant fugu obscurus.
Background
Takifugu obscurus is the first three delicacies in Yangtze river, is a fish with a long culture, is deeply loved by people in middle and lower reaches of Yangtze river, is a fish migrating in the river and the sea, and migrates to the sea when the temperature is 25-28 ℃ most suitable for growth and about 18 ℃ in 11 months every year, and stops eating and even dies at 12 ℃. Based on this characteristic of Fugu obscurus, it is difficult for Fugu obscurus to survive in natural conditions in winter in the middle and downstream regions of the Yangtze river. Therefore, the cultivation of the takifugu obscurus in winter needs to adopt a greenhouse for heating and keep the temperature at about 18 ℃ so as to ensure that the takifugu obscurus can successfully pass the winter.
However, the method for cultivating the fugu obscurus by heating in the greenhouse can only maintain the vital signs of the fugu obscurus though the fugu obscurus can overwinter, so that the fugu obscurus cultivated in the artificial greenhouse grows slowly, and the cultivation cost of the fugu obscurus in winter is also increased seriously.
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 for breeding the fugu obscurus 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 of:
s01, breeding the parent fish of the takifugu obscurus, which comprises the following specific operations: selecting the target adult fish with strong physique, good color, good gonad development and maximum specification from the population as the parent fish for propagation.
S02, inducing the fugu obscurus by adopting a low-temperature induction method in a gonadal development stage, wherein the method comprises the following specific operations: and (3) performing low-temperature stimulation on the fugu obscurus in the second stage and the third stage of the gonad development of the fugu obscurus selected in the step (S01).
S03, stimulating fertilized eggs of the takifugu obscurus in low-temperature stimulation development, wherein low-temperature stimulation is performed 30-120 min after the fertilized eggs of the takifugu obscurus, the stimulation temperature is 0-4 ℃, and the stimulation time is 30-120 seconds.
S04, selecting adult fugu obscurus fish with high growth speed by groups, and specifically operating as follows: and (3) hatching the fertilized eggs into fish fries, culturing the fish fries for about 6 months, selecting a preset culture pond from the beginning of 11 months to the beginning of 12 months when the water temperature of the fish pond is 12-15 ℃, selecting a preset number of adult fishes with high growth speed and maximum specification in each culture pond, transferring the adult fishes temporarily cultured in the culture pond in the greenhouse, and numbering each culture pond.
S05, cooling and screening the cold-resistant takifugu obscurus adult fish, which comprises the following specific operations: adjusting the culture water temperature, performing gradient cooling, and selecting adult fishes selected in S04 as small groups resistant to 12 ℃, 10 ℃, 8 ℃, 7 ℃, 6 ℃ and 5 ℃ according to tolerance temperature.
S06, excavating cold-resistant main effect genes, and specifically operating as follows: dividing the takifugu obscurus selected in the S05 into 6 groups according to tolerance temperature, randomly extracting a predetermined number of adult fishes from each group, performing gene sequencing, comparing and analyzing 2 groups of gene differences at 12 ℃ and 5 ℃ to find and determine 3-5 cold-resistant major effect genes, comparing and analyzing 4 groups of genes at 10 ℃, 8 ℃, 7 ℃ and 6 ℃ to find and determine the gene purity of the cold-resistant major effect genes in each group, and determining the cold-resistant major effect genes.
S07, establishing a cold-resistant 5 ℃ parent fish population, and specifically operating as follows: selecting adult fish with the homozygous cold-resistant main effect gene content of more than 3 groups as reserve parent fish.
S08. Propagating cold-resistant F 1 And performing cold resistance verification, wherein the specific operations are as follows: breeding fry to form adult fish according to a conventional method after the backup parent fish is sexually mature, 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 takifugu obscurus according to the cooling, and verifying F 1 Cold resistance of the synthetic fish.
Further, in the step S02, a method of inducing fugu obscurus at a low temperature in a gonadal development stage is used for breeding parent fish, and the specific operations are as follows: selecting basic parent fish with each half of male and female in a preset quantity, controlling the temperature reduction from 18 ℃, reducing the temperature by 1-2 ℃ each time, keeping the temperature for 24h, circularly reducing the temperature to 12 ℃, keeping the temperature for 48h, eliminating the inferior parent fish, recovering to the temperature above 18 ℃, and continuously culturing.
Further, in the step S03, breeding fertilized eggs of takifugu obscurus under low-temperature stimulation development specifically includes:
s0301, stimulating the first fertilized eggs, namely 10 ten thousand fertilized eggs, after 30 minutes of fertilization, soaking the fertilized eggs in water at 0 ℃ for 30 seconds, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching the fertilized eggs at normal temperature, breeding the fries in 3 fish ponds, and numbering the fish ponds;
s0302, stimulating the fertilized eggs of the second batch, soaking the fertilized eggs in 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 at normal temperature, breeding the fry in 3 fish ponds, and numbering the fish ponds;
s0303, stimulating fertilized eggs of the third batch, soaking the fertilized eggs in water at 0 ℃ for 120 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs into an incubation barrel, incubating the fertilized eggs at a normal temperature, breeding the fries in 3 fish ponds, and numbering the fish ponds;
s0304, stimulating the fourth batch of fertilized eggs, soaking the fertilized eggs in water at 4 ℃ for 60 seconds after fertilization for 60 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching at normal temperature, breeding the fry in 3 fish ponds, and numbering the fish ponds;
s0305, stimulating the fifth group of fertilized eggs, soaking the fertilized eggs in water at 4 ℃ for 90 seconds after fertilization for 90 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching at normal temperature, breeding the fry in 3 fish ponds, and numbering the fish ponds;
s0306, stimulating the sixth group of fertilized eggs, soaking the fertilized eggs in water at 4 ℃ for 120 seconds after fertilization for 120 minutes, taking out the fertilized eggs, transferring the fertilized eggs to a hatching barrel, hatching at normal temperature, breeding the fry in 3 fish ponds, and numbering the fish ponds.
Further, in the step S05, the cold-resistant takifugu obscurus adult fish is subjected to cooling screening, and the method is specifically characterized in that:
each time, the temperature is reduced by 1-2 ℃, each temperature gradient is kept for 24h, and the temperature is reduced to 5 ℃, so that the takifugu obscurus with poor cold resistance is eliminated.
Has the advantages that: the breeding method for breeding cold-resistant fugu obscurus provided by the invention has the following advantages:
1. the invention adopts low temperature to stimulate parent fish and fertilized egg in gonad development, improves the probability of gene mutation of egg cell sperm and fertilized egg in the development and division process, and is convenient for selecting excellent gene and character.
2. The invention excavates the cold-resistant major effect gene of the bred cold-resistant fugu obscurus, performs gene sequencing, compares gene difference, finds and determines the cold-resistant major effect gene and the cold-resistant major effect gene, and facilitates the breeding of the cold-resistant fugu obscurus at the later stage.
3. The fugu obscurus bred by the breeding method has the cold-resistant characteristic, so that the winter breeding efficiency of the fugu obscurus under natural conditions is improved, and the cost for breeding the fugu obscurus in winter is further reduced.
Detailed Description
The present invention will be further described with reference to the following 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 of:
s01, selecting and breeding parent fishes of takifugu obscurus, which comprises selecting target adult fishes meeting the requirements of strong physique, good color, good gonad development and maximum specification from groups as the bred parent fishes.
S02, inducing the Fugu obscurus by adopting a gonad development stage low-temperature induction method, stimulating the Fugu obscurus at low temperature in the second stage and the third stage of the gonad development of the Fugu obscurus selected in S01, selecting 2000 basic parent fishes of each half of male and female, controlling and cooling from 18 ℃, cooling for 1 ℃ for the first time, keeping the temperature for 24 hours, cooling for 1.5 ℃ for the second time, keeping the temperature for 24 hours, cooling for 2 ℃ for the third time, keeping the temperature for 24 hours, cooling for 1.5 ℃ for the fourth time, namely cooling to 12 ℃, keeping the temperature for 48 hours, eliminating the Fugu obscurus with poor cold resistance, recovering to the temperature of more than 18 ℃, and continuing cultivation.
S03, stimulating the developing fish germ cells at low temperature, wherein low-temperature stimulation is carried out 30-120 min after the fish germ cells are fertilized, the stimulation temperature is 0-4 ℃, and the stimulation time is 30-120 seconds.
S0301, stimulating the first fertilized eggs, soaking the fertilized eggs in water at 0 ℃ for 30 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs into an incubation barrel, incubating according to normal temperature, breeding the fry in 3 fish ponds, and numbering the fish ponds.
S0302, stimulating the fertilized eggs of the second batch, soaking the fertilized eggs in water at 0 ℃ for 60 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs to a hatching barrel, hatching at normal temperature, breeding the fry in 3 fish ponds, and numbering the fish ponds.
S0303, stimulating the fertilized eggs of the third batch, namely 10 thousands of fertilized eggs, after 30 minutes of fertilization, soaking the fertilized eggs in water at 0 ℃ for 120 seconds, taking out the fertilized eggs, transferring the fertilized eggs to a hatching barrel, hatching the fertilized eggs according to normal temperature, and breeding the fry in 3 fish ponds, wherein the fish ponds are numbered.
S0304, stimulating the fourth batch of fertilized eggs, soaking the fertilized eggs in water of 4 ℃ for 60 seconds after fertilization for 60 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching at normal temperature, breeding the fry in 3 fish ponds, and numbering the fish ponds.
S0305, stimulating the fifth group of fertilized eggs, namely 10 thousands of fertilized eggs, after 90 minutes of fertilization, soaking the fertilized eggs in water at 4 ℃ for 90 seconds, taking out the fertilized eggs, transferring the fertilized eggs to a hatching barrel, hatching the fertilized eggs according to normal temperature, and breeding the fries in 3 fish ponds, wherein the fish ponds are numbered.
S0306, stimulating the sixth group of fertilized eggs, namely 10 thousands of fertilized eggs, after fertilization for 120 minutes, soaking the fertilized eggs in water at 4 ℃ for 120 seconds, taking out the fertilized eggs, transferring the fertilized eggs to a hatching barrel, hatching the fertilized eggs according to normal temperature, and breeding the fries in 3 fish ponds, wherein the fish ponds are numbered.
S04, selecting groups of adult fishes of the takifugu obscurus with high production speed, hatching fertilized eggs of the S03 into fish fries, culturing the fish fries for about 6 months, selecting 18 culture ponds of the S03 when the water temperature of the fish ponds is 12-15 ℃ at the beginning of 11-12 months, selecting 1000 fish with high production speed and maximum specification 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 and screening cold-resistant adult takifugu obscurus, adjusting the culture water temperature to 12 ℃, keeping the culture water temperature for 24 hours, selecting the adult takifugu obscurus which is resistant to 12 ℃, and numbering the adult takifugu obscurus; continuously adjusting the culture water temperature to 10 ℃, keeping the temperature for 24 hours, selecting adult fishes resistant to 10 ℃, and numbering the adult fishes; continuously adjusting the culture water temperature to 8 ℃, keeping the culture water temperature for 24 hours, selecting adult fishes resistant to 8 ℃, and numbering the adult fishes; continuously adjusting the culture water temperature to 7 ℃, keeping the temperature for 24h, selecting adult fishes resistant to 7 ℃, and numbering the adult fishes; continuously adjusting the culture water temperature to 6 ℃, keeping the temperature for 24 hours, selecting adult fishes resistant to 6 ℃, and numbering the adult fishes; continuously adjusting the culture water temperature to 5 ℃, keeping the culture water temperature for 24 hours, selecting adult fishes resistant to 5 ℃, and numbering the adult fishes.
S06, excavating cold-resistant major effect genes, randomly extracting 6 groups of groups selected in the S05, selecting 50 fish tails from each group of groups, carrying out gene sequencing, comparing and analyzing 2 groups of gene differences at 12 ℃ and 5 ℃, finding and determining 3-5 cold-resistant major effect genes, comparing and analyzing 4 groups of genes at 10 ℃, 8 ℃, 7 ℃ and 6 ℃, finding and determining the gene purity degree in each group of the cold-resistant major effect genes, and determining the cold-resistant major effect genes.
S07, establishing a cold-resistant 5 ℃ parent fish population, and selecting adult fishes with the content of homozygous cold-resistant main effect genes being more than 3 groups as backup parent fishes.
S08. Propagating cold-resistant F 1 And a step F of verifying the cold resistance, namely breeding fries to adult fish according to a conventional method after the backup parent fish is sexually mature, 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 1 Cold resistance of the synthetic fish.
The bred adult fugu obscurus fish lay eggs in 4-5 months, the fry is cultured to 12-5 ℃, and simultaneously, the bred fugu obscurus and the unseeded fugu obscurus are sampled, measured and analyzed, wherein the average weight of the bred fugu obscurus is 249.6 +/-50.12 g, and the bred fugu obscurus still has full vitality under the environment of 5 ℃; the average weight of the unseeded fugu obscurus is 110.3 +/-28.93 g, most of the fugu obscurus stops eating food at 18 ℃ in the early 11 months and gradually dies, and all the fugu obscurus dies at 5 ℃. The result shows that the bred takifugu obscurus has obvious cold resistance.
The embodiments described herein are only a part of the embodiments of the present invention, and not all embodiments, and all other embodiments obtained by those skilled in the art based on the embodiments of the present invention without any inventive work belong to the protection 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, selecting parent fishes of fugu obscurus, wherein the selection comprises the step of selecting target adult fishes which meet the requirements of strong physique, good color, good gonad development and maximum specification from groups as the bred parent fishes;
s02, inducing the fugu obscurus by adopting a low-temperature induction method in the gonadal development stage, wherein the low-temperature stimulation is carried out on the fugu obscurus in the second stage and the third stage of the gonadal development of the fugu obscurus selected in the S01;
s03, stimulating fertilized eggs of the takifugu obscurus in the low-temperature stimulation development, wherein the low-temperature stimulation is carried out 30-120 min after the fertilized eggs of the takifugu obscurus are fertilized, the stimulation temperature is 0-4 ℃, and the stimulation time is 30-120 seconds;
s04, selecting adult fugu obscurus fish with high growth speed by groups, and specifically operating as follows: the fertilized eggs are hatched into fish fries in the step S03, the fish fries are cultured for about 6 months, a preset culture pond is selected when the water temperature of the fish pond is 12-15 ℃ from the beginning of 11 months to the beginning of 12 months, a preset number of fish with high growth speed and maximum specification are selected in each culture pond, the culture ponds temporarily cultured in the greenhouse are transferred, and each culture pond is numbered;
s05, cooling and screening the cold-resistant takifugu obscurus adult fish, which comprises the following specific operations: adjusting the culture water temperature, performing gradient cooling, and selecting the adult fugu obscurus selected in S04 as a small population tolerant to 12 ℃, 10 ℃, 8 ℃, 7 ℃, 6 ℃ and 5 ℃ according to tolerance temperature;
s06, excavating cold-resistant main effect genes, and specifically operating as follows: dividing the takifugu obscurus adult fish selected in the S05 into 6 groups, randomly extracting a predetermined number of takifugu obscurus from each group, performing gene sequencing, comparing and analyzing 2 groups of gene differences at 12 ℃ and 5 ℃ to find and determine 3-5 cold-resistant major effect genes, comparing and analyzing 4 groups of genes at 10 ℃, 8 ℃, 7 ℃ and 6 ℃ to find and determine the gene purity degree in each group of the cold-resistant major effect genes, and determining the cold-resistant major effect genes;
s07, establishing a cold-resistant 5 ℃ parent fish population, and specifically operating as follows: selecting adult fishes of the takifugu obscurus with homozygous cold-resistant major effect gene content of more than 3 groups as backup parent fishes;
s08, propagating cold-resistant F 1 And instead, carrying out cold resistance verification, and specifically operating as follows: breeding fry to adult fish according to a conventional method after the backup parent fish is sexually mature, randomly selecting a predetermined number of fish ponds, selecting a predetermined number of adult fish in each pond, cooling and screening the breeding method of the cold-resistant adult fish according to the step S05, and verifying F 1 Cold resistance of the synthetic fish.
2. The method for breeding cold-resistant Fugu obscurus through low-temperature induction according to claim 1, wherein the step S02 of inducing Fugu obscurus through the gonadal development stage low-temperature induction method comprises the following specific operations: selecting basic parent fish with each half of male and female in a preset quantity, controlling the temperature reduction from 18 ℃, reducing the temperature by 1-2 ℃ each time, keeping the temperature for 24h, circularly reducing the temperature to 12 ℃, keeping the temperature for 48h, eliminating the inferior parent fish, recovering to the temperature above 18 ℃, and continuously culturing.
3. The method for breeding cold-resistant Fugu obscurus through low-temperature induction as claimed in claim 1, wherein the step S03, the breeding of fertilized eggs of Fugu obscurus through low-temperature stimulation comprises:
s0301, stimulating the first fertilized eggs, soaking the fertilized eggs in water at 0 ℃ for 30 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs into an incubation barrel, incubating according to a normal temperature, breeding the fry in 3 fish ponds, and numbering the fish ponds;
s0302, stimulating the fertilized eggs of the second batch, soaking the fertilized eggs in 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 at normal temperature, breeding the fry in 3 fish ponds, and numbering the fish ponds;
s0303, stimulating a third batch of fertilized eggs, soaking the fertilized eggs in water at 0 ℃ for 120 seconds after fertilization for 30 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching at normal temperature, breeding the fry in 3 fish ponds, and numbering the fish ponds;
s0304, stimulating the fourth batch of fertilized eggs, soaking the fertilized eggs in water at 4 ℃ for 60 seconds after fertilization for 60 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching at normal temperature, breeding the fry in 3 fish ponds, and numbering the fish ponds;
s0305, stimulating the fifth batch of fertilized eggs, soaking the fertilized eggs in water at 4 ℃ for 90 seconds after fertilization for 90 minutes, taking out the fertilized eggs, transferring the fertilized eggs into a hatching barrel, hatching at normal temperature, breeding the fry in 3 fish ponds, and numbering the fish ponds;
and S0306, stimulating the sixth group of fertilized eggs, soaking the fertilized eggs in water of 4 ℃ for 120 seconds after fertilization for 120 minutes, taking out the fertilized eggs, transferring the fertilized eggs to a hatching barrel, hatching at normal temperature, breeding the fries in 3 fish ponds, and numbering the fish ponds.
4. The method for low-temperature induced breeding of cold-resistant Fugu obscurus according to claim 1, wherein in the step S05, the step of cooling and screening the cold-resistant Fugu obscurus is characterized in that:
each time, the temperature is reduced by 1-2 ℃, each temperature gradient is kept for 24h, and the temperature is reduced to 5 ℃, so that the takifugu obscurus with poor cold resistance is eliminated.
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| 李文豪;冉皓宇;陈良标;: "低温胁迫导致斑马鱼尾部发育畸形的分子机制", 基因组学与应用生物学, no. 06, pages 55 - 60 * |
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