CN106614142A - Method for producing sterile hybrid nibea albiflora - Google Patents
Method for producing sterile hybrid nibea albiflora Download PDFInfo
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- CN106614142A CN106614142A CN201611233683.0A CN201611233683A CN106614142A CN 106614142 A CN106614142 A CN 106614142A CN 201611233683 A CN201611233683 A CN 201611233683A CN 106614142 A CN106614142 A CN 106614142A
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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
- A01K61/00—Culture of aquatic 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
- 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
Abstract
The invention discloses a method for producing sterile hybrid nibea albiflora. The method includes the following steps: using nibea mitsukurii as a female parent and pennahia argentatus as a male parent, and promoting the sexual glands of the nibea mitsukurii and the pennahia argentatus to be mature synchronously; selecting the nibea mitsukurii with well-developed sexual glands and the pennahia argentatus with well-developed sexual glands, and injecting lutein release promoting hormones for artificially induced spawning of the nibea mitsukurii; collecting an ovum of the nibea mitsukurii and a sperm of the pennahia argentatus, and conducting dry fertilization; breeding hybrid larva fish, hybrid young fish and hybrid juvenile fish respectively, and observing the sexual gland. The hybridization method is used for culturing the nibea albiflora with the sterile sexual gland, and the bred nibea albiflora has the advantages of being high in survival rate, strong in stress resistance, totally sterile and the like. The method has the advantages of being simple and easy to operate, and the sterile hybrid nibea albiflora has a good application prospect in aquaculture production, can be used as an artificial 'surrogate' receptor, and has very important application value.
Description
Technical field
The present invention relates to technical field of fishery, the method for specifically producing sterile spotted maigre using the method for artificial hybridization,
More particularly, to a kind of production method of cross-sterility spotted maigre.
Background technology
The fish of sexual gland infertility have higher scientific research and using value in aquaculture.First, the fish of sexual gland infertility
Class often shows the advantages such as fast, the dressing percentage height of growth；Secondly, the fish of sexual gland infertility can be reduced because fish escape into nature
The ecological risk that boundary brings；Furthermore, the fish of sexual gland infertility have special application in scientific research, such as in Technique in Fishes " replace-conceive "
In research, the fish of sexual gland infertility produce the sperm or ovum of other fingerlings as acceptor.Therefore, explore and produce sexual gland infertility fish
The method of class is significant in aquaculture.
Generally, artificial hybridization, triploid induction, gene knockout can be passed through（Drop）Etc. the fish that method produces sexual gland infertility
Class.Wherein, artificial hybridization is used as one of classical breeding method, due to easy to operate and most attention that be subject to researcher.People
Work hybridization is to carry out hybridization using the fingerling between different inter-species, category or between section to produce offspring.Artificial hybridization success or not, Hou Daishi
No to survive or infertility, depending on the compatibility of the Parent inhereditary material for hybridization, the result of hybridization is often difficult pre-
Survey.Therefore, obtaining the filial generation of infertility completely needs substantial amounts of experiment sieving, checking.We make spotted maigre by dustpan
(Nibea mitsukurii), spotted maigre (Nibea albiflora), Yin Guyu (Pennahia argentata), Japan it is yellow
Aunt fish (Argyrosomus japonicus), large yellow croaker (Larimichthys crocea), little yellow croaker (Larimichthys polyactis) etc. carry out the cross experiment of multiple combinations, the cross combination that dustpan makees spotted maigre and silver-colored aunt fish is filtered out, after hybridization
In generation, can be completely sterile, has preferable application prospect in breeding production, and can be used as the acceptor of artificial " replace-conceive ", tool
There is higher research application value.
The content of the invention
It is an object of the invention to provide it is a kind of have be easy to execute-in-place, safe and reliable, simple, survival rate is high, anti-
Inverse property is strong, the method that infertility completely utilization artificial hybridization method produces sexual gland infertility spotted maigre.
The technical scheme adopted by the invention for realizing the object of the invention is：A kind of production method of cross-sterility spotted maigre,
The method is comprised the following steps：
（1）The cultivation of parent population
Select dustpan to make spotted maigre raun and silver-colored aunt fish milter as parent population, make spotted maigre to dustpan respectively and silver-colored aunt fish to carry out nutrition strong
Change and the regulation and control of light temperature, promote its sexual gland synchronous ripe；Cultivated using heating up during cultivation, water temperature is gradually raised to 24 DEG C by 18 DEG C；
The preferable parent of gonad development is selected in parent fish rearing pond, the parent after selecting is placed in black cloth and covers in bucket, keep cultivating water temperature
For 22-24 DEG C；
（2）Female dustpan makees the artificial induced spawning of spotted maigre
Female dustpan is hastened parturition as spotted maigre, from injection Luteinizing hormone releasing-hormone A2 as ocyodinic, injection dosage
1.5-2 μ g oxytocins are injected for 1kg parents, the ecbolic effect time is 28-32h when the cultivation temperature of parent is 24 DEG C；To dustpan
Make spotted maigre and ovum is adopted using manual compression belly method；
（3）Artificial collection silver aunt's milt liquid
Silver-colored aunt's milt liquid is gathered using manual compression belly method, with sterilized dry glass dropper clean seminal fluid is drawn, use Jing
The dry glass dropper of sterilization draws clean seminal fluid, in being stored in 1.5 mL centrifuge tubes, is placed on ice；
（4）Dustpan makees spotted maigre ♀ × silver aunt's fish ♂ artificial hybridization
Dustpan is made into fish-egg of the spotted maigre using the collection of manual compression belly method, is put into and is dried in beaker；By the silver-colored aunt fish of collection
Fresh semen is instilled in unfertilized fish-egg, dry method fertilization, it is fully mixed with appropriate sea-water activated rear agitation, is subsequently adding
The seawater of 2-5 times of ovum amount volumes completes fertilization process；
（5）Incubating oosperm
Embryonated egg is put into 500L glass flumes and is hatched, water temperature is 24 DEG C, and salinity 28～32 ‰, pH8.0～8.2, hydrostatic is micro-
Inflation；
（6）The cultivation of cross hybrid seedling
Carry out the cultivation of cross hybrid seedling after incubating oosperm, cultivation is divided into alevin stage, postlarva phase, young stage, cultivate water temperature be 20～
24 DEG C, salinity 28～32 ‰, pH8.0～8.2, quantity of exchanged water and aeration quantity gradually increase with fry growth；Seed rearing is adopted
Bait be followed successively by wheel animalcule, artemia nauplii, mixed feed；
（7）The success rate of inspection cross hybrid seedling infertility
When cross hybrid seedling was cultivated to 3 monthly age, solution takes sexual gland, is fixed using Bonn formula liquid, and using histotomy sterile feelings are observed
Condition.
The present invention cultivates the spotted maigre of infertility using the method for hybridization, and the spotted maigre cultivated has survival rate high, degeneration-resistant
Property it is strong, completely infertility the features such as, the method has the advantages that simple, and the sterile spotted maigre cultivated has in breeding production
There is preferable popularizing application prospect, and can be as the acceptor of artificial " replace-conceive ", with very important using value.
Preferably, step（1）In the cultivation of parent population, dustpan makees spotted maigre and silver-colored aunt fish cultivates respectively in 1 ton of tank, often
Individual tank cultivates 30-40 tails, and using sand filter seawater miniflow water is continued, and 1 time is fed daily；Tank plastic covering canopy is incubated, water temperature
More than 18 DEG C are maintained at, when water temperature is less than 18 DEG C, using heating rod heating maintenance water temperature more than 18 DEG C；The 1m above tank
The fluorescent lamp of 1 60W is installed at place, makes light be evenly distributed in the water surface of culture pond, and intensity of illumination is 50～100Lx；Parent population is urged
Antenatal 4 weeks, water temperature raised 1 DEG C every 5 days, gradually water temperature was risen into 24 DEG C by 18 DEG C；Light intensity increased by 1 hour every 5 days.
Preferably, step（2）Female dustpan is made in the artificial induced spawning of spotted maigre, and concrete operations are：Make yellow aunt in female dustpan
Fish carries out hastening parturition after 28h, spotted maigre is made to dustpan and adopts ovum using manual compression belly method, and both hands are used fish belly portion both sides are simultaneously uniform
Power, from rear and front extruding belly slowly, flows out ovum, and using 500mL glass beakers ovum is accessed, and ovum is put into dry
In beaker.
Preferably, step（6）The cultivation of cross hybrid seedling：The cultivation of alevin stage, 1～15 age in days adopts the cultivation that adds water, daily
Add water as the 10%～20% of water, cultivate density 2～30,000 tails/m3, faint inflation, water temperature is maintained at 24 DEG C；
The cultivation of postlarva phase, 15～25 ages in days add water daily twice, and each quantity of exchanged water is the 50%～100% of water, cultivate density
0.5～10,000 tails/m3, per two days soil pick-ups 1 time, 24 DEG C of culture pond water temperature；
The cultivation of young stage, after 25 ages in days, changes daily water once, quantity of exchanged water 100%, cultivates density depending on fish body size, cultivates
Temperature is nature seawater temperature.
Preferably, step（4）In the cultivation of cross hybrid seedling, the bait that fry rearing is adopted is followed successively by wheel animalcule, artemia without section
The young, mixed feed；Feeding method is：The age in days not bait throwing in of fry 1～2,3～14 ages in days feed wheel animalcule, and the age in days of newly hatched larvae 3 is opened
Mouth is ingested, and open-mouthed bait is brachionus plicatilis, is strengthened more than 12 hours with high concentration chlorella before feeding；15～20 ages in days are thrown
Wheel animalcule and artemia nauplii are fed, 20～25 ages in days feed artemia nauplii and mixed feed, the wheel animalcule feeds density
For 20～40/mL, the density that feeds of the artemia nauplii is 5～15/mL；26 ages in days start domestication and feed cooperation
Feed, 31 ages in days feed completely mixed feed, and domestication tames mixed feed after success using a small amount of multiple feeding method
Feeding volume regards the alimentary canal turgor of fry and observation bottom of pond residual bait situation is determined.
Preferably, step（7）In the success rate of inspection cross hybrid seedling infertility, when cross hybrid seedling was cultivated to 3 monthly age, randomly select
30 tails, take sexual gland after dissection, sexual gland presents thin-line-shaped；Sexual gland is fixed to be changed after 12h using Bonn formula liquid and is protected using 70% alcohol
Deposit；Will the dehydration of sexual gland sample, carry out histotomy after FFPE, the Gonadal Morphology after section without spermary lobule or ovarian structure,
Therefore it is accredited as sexual gland infertility.
Advantages of the present invention and good effect are：The present invention on the basis of repetition test, filter out dustpan make spotted maigre and
The cross combination of silver-colored aunt fish, filial generation can be completely sterile, so as to set up a kind of method of extensive cultivation infertility spotted maigre,
The method has the advantages that to be easy to execute-in-place, safe and reliable, simple.The sterile spotted maigre cultivated is in breeding production
With preferable popularizing application prospect, and can be as the acceptor of artificial " replace-conceive ", with very important using value.
Description of the drawings
Fig. 1 is a kind of structural representation that three monthly ages hybridized spotted maigre in the present invention;
Fig. 2 is a kind of structural representation that three monthly ages hybridized spotted maigre sexual gland in the present invention;
Fig. 3 is to hybridize the section of spotted maigre gonadal tissue in the present invention.
Specific embodiment
Below by specific embodiment and combine accompanying drawing technical scheme is described in further detail.
Embodiment 1：
A kind of breeding method of sexual gland infertility spotted maigre, the method is comprised the following steps：
（1）The cultivation of parent population
Select dustpan to make spotted maigre raun and silver-colored aunt fish milter as parent population, make spotted maigre to dustpan respectively and silver-colored aunt fish to carry out nutrition strong
Change and the regulation and control of light temperature, promote its sexual gland synchronous ripe；Cultivated using heating up during cultivation, water temperature is gradually raised to 24 DEG C by 18 DEG C；
The preferable parent of gonad development is selected in parent fish rearing pond, the parent after selecting is placed in black cloth and covers in bucket, keep cultivating water temperature
For 22-24 DEG C；
（2）Female dustpan makees the artificial induced spawning of spotted maigre
Female dustpan is hastened parturition as spotted maigre, from injection Luteinizing hormone releasing-hormone A2 as ocyodinic, injection dosage
1.5-2 μ g oxytocins are injected for 1kg parents, the ecbolic effect time is 28-32h when the cultivation temperature of parent is 24 DEG C；To dustpan
Make spotted maigre and ovum is adopted using manual compression belly method；
（3）Artificial collection silver aunt's milt liquid
Silver-colored aunt's milt liquid is gathered using manual compression belly method, with sterilized dry glass dropper clean seminal fluid is drawn, use Jing
The dry glass dropper of sterilization draws clean seminal fluid, in being stored in 1.5 mL centrifuge tubes, is placed on ice；
（4）Dustpan makees spotted maigre ♀ × silver aunt's fish ♂ artificial hybridization
Dustpan is made into fish-egg of the spotted maigre using the collection of manual compression belly method, is put into and is dried in beaker；By the silver-colored aunt fish of collection
Fresh semen is instilled in unfertilized fish-egg, dry method fertilization, it is fully mixed with appropriate sea-water activated rear agitation, is subsequently adding
The seawater of 5 times of ovum amount volumes completes fertilization process；
（5）Incubating oosperm
Embryonated egg is put into 100L glass flumes and is hatched, water temperature is 24 DEG C, salinity 28～32 ‰, pH 8.0～8.2, and hydrostatic is micro-
Inflation；
（6）The cultivation of cross hybrid seedling
Carry out the cultivation of cross hybrid seedling after incubating oosperm, cultivation is divided into alevin stage, postlarva phase, young stage, cultivate water temperature be 20～
24 DEG C, salinity 28～32 ‰, pH8.0～8.2, quantity of exchanged water and aeration quantity gradually increase with fry growth；Seed rearing is adopted
Bait be followed successively by wheel animalcule, artemia nauplii, mixed feed；
（7）The success rate of inspection cross hybrid seedling infertility
When cross hybrid seedling was cultivated to 3 monthly age, solution takes sexual gland, is fixed using Bonn formula liquid, and using histotomy sterile feelings are observed
Condition.
Concrete operations are as follows：
（1）Dustpan makees spotted maigre and silver-colored aunt fish is cultivated respectively in 1 ton of tank, and each tank cultivates 30-40 tails, using husky filter sea
Water continues miniflow water, and 1 time is fed daily.Tank plastic covering canopy is incubated, and water temperature is maintained at more than 18 DEG C, when water temperature is less than 18 DEG C
When, using heating rod heating maintenance water temperature more than 18 DEG C.The fluorescent lamp of 1 60W is installed at 1m above tank, lamp is made as far as possible
Light is evenly distributed in the water surface of culture pond, and intensity of illumination is 50～100Lx.Parent population hasten parturition before 4 weeks, water temperature every 5d raise 1 DEG C,
Gradually water temperature is risen into 24 DEG C by 18 DEG C；Light intensity increases 1h every 5d.
（2）The preferable parent of gonad development is selected in parent fish rearing pond carries out artificial induced spawning, and the parent after selecting is placed in
In the tank that black cloth is covered.Choose 10 tails female dustpan to be hastened parturition as spotted maigre, what ocyodinic was selected is Ningbo the second hormone factory
The injection Luteinizing hormone releasing-hormone A2 of production, dilution is prepared with compound amino acid solution, and injection dosage is 1kg parents note
Penetrate 1.5-2 μ g oxytocins.
（3）Hasten parturition after 28h, spotted maigre is made to dustpan ovum is adopted using manual compression belly method, concrete operations are：With wet from sea water
Towel parcel fish body, expose abdominal reproductive hole position and with absorbent cotton by gonopore and surrounding wiped clean, fix parent population
Afterwards, both hands are simultaneously uniform firmly in fish belly portion both sides, from rear and front extruding belly slowly, that is, have ovum to flow out, using 500mL
Glass beaker accesses ovum, and ovum is put into dry beaker.10 tail dustpans have 8 acquisition ovums, success rate of hastening parturition as spotted maigre
80%.Using the same silver-colored aunt's milt liquid of manual compression belly method collection, with sterilized dry glass dropper clean essence is drawn
Liquid, is stored in 1.5 mL centrifuge tubes and is placed on ice, seminal fluid being added in unfertilized egg（Seminal fluid about 0.5～1mL）, dry method receives
Both are gently shaken mixing by essence, and the seawater for being subsequently adding 5 times of ovum amount volumes completes fertilization process.
（4）Incubating oosperm
Embryonated egg is put into 500L glass flumes and is hatched, water temperature is 24 DEG C, and salinity 28～32 ‰, pH8.0～8.2, hydrostatic is micro-
Inflation.
（5）The cultivation of cross hybrid seedling
The cultivating process of cross hybrid seedling is divided into alevin stage (1～15 age in days), postlarva phase (15～25 age in days) and young stage（25 ages in days with
Afterwards）.Alevin stage, using adding water cultivations, adds water daily as the 10% ~ 20% of water, cultivation density 2 ~ 30,000 tails/m3, faint inflation, water
Temperature is maintained at 24 DEG C.
The postlarva phase（15～25 ages in days）, it is that twice, each quantity of exchanged water is the 50% ~ 100% of water to add daily, cultivates density
0.5 ~ 10,000 tails/m3, per two days soil pick-ups 1 time, 24 DEG C of culture pond water temperature.
Young stage（After 25 ages in days）, water is changed daily once, quantity of exchanged water 100% cultivates density depending on fish body size, cultivates
Temperature is nature seawater temperature.
The salinity that alevin stage and postlarva phase cultivate is 28～32 ‰, pH8.0～8.2.
The bait that fry rearing is adopted is followed successively by wheel animalcule, artemia nauplii, mixed feed.Feeding method is：Fry 1 ~ 2
Age in days not bait throwing in, 3 ~ 14 ages in days feed wheel animalcule, the age in days initial feeding of newly hatched larvae 3, and open-mouthed bait is brachionus plicatilis, is fed
Front use high concentration chlorella is strengthened more than 12 hours；15 ~ 20 ages in days feed wheel animalcule and artemia nauplii, and 20 ~ 25 ages in days feed
Artemia nauplii and mixed feed, the wheel animalcule feeds density for 20 ~ 40/mL, and the artemia nauplii feeds
Density is 5 ~ 15/mL；26 ages in days start domestication and feed mixed feed, and 31 ages in days feed completely mixed feed, and domestication is adopted
With a small amount of multiple feeding method, alimentary canal turgor and observation pond of the feeding volume of mixed feed after success regarding fry is tamed
Bottom residual bait situation is determined.
（6）When cross hybrid seedling was cultivated to 3 monthly age, 30 tails are randomly selected, sexual gland is taken after dissection, sexual gland presents thin-line-shaped（See
Fig. 1, Fig. 2）.Sexual gland is fixed to be changed after 12h using Bonn formula liquid and is preserved using 70% alcohol.By the dehydration of sexual gland sample, FFPE
After carry out histotomy, the Gonadal Morphology after section without spermary lobule or ovarian structure, therefore be accredited as sexual gland infertility（See Fig. 2）.
The production method of the cross-sterility spotted maigre, makees spotted maigre as female parent with dustpan respectively, and silver-colored aunt fish is male parent, promotes it
Sexual gland is synchronous ripe；Select the good dustpan of gonad development and make spotted maigre and silver-colored aunt's fish parent fish, injection luteinizing hormone releasing hormone
A2 carries out artificial induced spawning to dustpan as spotted maigre；Collection dustpan makees spotted maigre ovum and silver-colored aunt's milt, carries out dry method fertilization；Hybridization
Larval and juvenile fish is cultivated and sexual gland observation.The spotted maigre of sexual gland infertility is cultivated using the method for hybridization, the spotted maigre tool cultivated
The features such as having survival rate height, strong stress resistance, complete infertility, the method has the advantages that simple, to cultivate sexual gland infertility
There is preferable popularizing application prospect in breeding production, and can be as the acceptor of artificial " replace-conceive ", with weighing very much
The using value wanted.
Claims (6)
1. a kind of production method of cross-sterility spotted maigre, it is characterised in that the method is comprised the following steps：
（1）The cultivation of parent population
Select dustpan to make spotted maigre raun and silver-colored aunt fish milter as parent population, make spotted maigre to dustpan respectively and silver-colored aunt fish to carry out nutrition strong
Change and the regulation and control of light temperature, promote its sexual gland synchronous ripe；Cultivated using heating up during cultivation, water temperature is gradually raised to 24 DEG C by 18 DEG C；
The preferable parent of gonad development is selected in parent fish rearing pond, the parent after selecting is placed in black cloth and covers in bucket, keep cultivating water temperature
For 22-24 DEG C；
（2）Female dustpan makees the artificial induced spawning of spotted maigre
Female dustpan is hastened parturition as spotted maigre, from injection Luteinizing hormone releasing-hormone A2 as ocyodinic, injection dosage
1.5-2 μ g oxytocins is injected for 1kg parents, the ecbolic effect time is 28-32h when the cultivation temperature of parent is 24 DEG C；Dustpan is made
Spotted maigre adopts ovum using manual compression belly method；
（3）Artificial collection silver aunt's milt liquid
Silver-colored aunt's milt liquid is gathered using manual compression belly method, with sterilized dry glass dropper clean seminal fluid is drawn, preserved
In 1.5 mL centrifuge tubes, it is placed on ice；
（4）Dustpan makees spotted maigre ♀ × silver aunt's fish ♂ artificial hybridization
Dustpan is made into fish-egg of the spotted maigre using the collection of manual compression belly method, is put into and is dried in beaker；By the silver-colored aunt fish of collection
Fresh semen is instilled in unfertilized fish-egg, dry method fertilization, it is fully mixed with appropriate sea-water activated rear agitation, is subsequently adding
The seawater of 2-5 times of ovum amount volume completes fertilization process；
（5）Incubating oosperm
Embryonated egg is put into 500L glass flumes and is hatched, water temperature is 24 DEG C, and salinity 28～32 ‰, pH8.0～8.2, hydrostatic is micro-
Inflation；
（6）The cultivation of cross hybrid seedling
Carry out the cultivation of cross hybrid seedling after incubating oosperm, cultivation is divided into alevin stage, postlarva phase, young stage, cultivate water temperature be 20～
24 DEG C, salinity 28～32 ‰, pH8.0～8.2, quantity of exchanged water and aeration quantity gradually increase with fry growth；Seed rearing is adopted
Bait be followed successively by wheel animalcule, artemia nauplii, mixed feed；
（7）The success rate of inspection cross hybrid seedling infertility
When cross hybrid seedling was cultivated to 3 monthly age, solution takes sexual gland, is fixed using Bonn formula liquid, and using histotomy sterile feelings are observed
Condition.
2. the production method of a kind of cross-sterility spotted maigre according to claim 1, it is characterised in that：Step（1）Parent population
Cultivation in, dustpan makees spotted maigre and silver-colored aunt fish is cultivated respectively in 1 ton of tank, each tank cultivation 30-40 tail, using husky filter sea
Water continues miniflow water, and 1 time is fed daily；Tank plastic covering canopy is incubated, and water temperature is maintained at more than 18 DEG C, when water temperature is less than 18 DEG C
When, using heating rod heating maintenance water temperature more than 18 DEG C；The fluorescent lamp of 1 60W is installed at 1m above tank, makes light exist
The water surface of culture pond is evenly distributed, and intensity of illumination is 50～100Lx；Parent population hasten parturition before 4 weeks, water temperature every 5 days raise 1 DEG C, by
Gradually water temperature is risen into 24 DEG C by 18 DEG C；Light intensity increased by 1 hour every 5 days.
3. the production method of a kind of cross-sterility spotted maigre according to claim 1, it is characterised in that：Step（2）Female
Dustpan is made in the artificial induced spawning of spotted maigre, and concrete operations are：After female dustpan carries out hastening parturition 28h as spotted maigre, spotted maigre is made to dustpan
Ovum is adopted using manual compression belly method, both hands are simultaneously uniform firmly in fish belly portion both sides, from rear and front extruding belly slowly, make ovum
Subflow goes out, and using 500mL glass beakers ovum is accessed, and ovum is put into dry beaker.
4. the production method of a kind of cross-sterility spotted maigre according to claim 1, it is characterised in that：Step（6）Hybridization
The cultivation of seedling：The cultivation of alevin stage, 1～15 age in days, using adding water cultivations, adds water daily as the 10%～20% of water, and cultivation is close
Degree 2～30,000 tails/m3, faint inflation, water temperature is maintained at 24 DEG C；
The cultivation of postlarva phase, 15～25 ages in days add water daily twice, and each quantity of exchanged water is the 50%～100% of water, cultivate density
0.5～10,000 tails/m3, per two days soil pick-ups 1 time, 24 DEG C of culture pond water temperature；
The cultivation of young stage, after 25 ages in days, changes daily water once, quantity of exchanged water 100%, cultivates density depending on fish body size, cultivates
Temperature is nature seawater temperature.
5. the production method of a kind of cross-sterility spotted maigre according to claim 1, it is characterised in that：Step（4）Hybridization
In the cultivation of seedling, the bait that fry rearing is adopted is followed successively by wheel animalcule, artemia nauplii, mixed feed；Feeding method is：Fry
1～2 age in days not bait throwing in, 3～14 ages in days feed wheel animalcule, the age in days initial feeding of newly hatched larvae 3, and open-mouthed bait is fold arm tail wheel
Worm, is strengthened more than 12 hours before feeding with high concentration chlorella；15～20 ages in days feed wheel animalcule and artemia nauplii, and 20～25
Age in days feeds artemia nauplii and mixed feed, and the density that feeds of the wheel animalcule is 20～40/mL, and the artemia is without section children
The density that feeds of body is 5～15/mL；26 ages in days start domestication and feed mixed feed, and 31 ages in days feed completely mixed feed,
Domestication regards the alimentary canal turgor of fry using a small amount of multiple feeding method, the feeding volume for taming mixed feed after success
Determine with observation bottom of pond residual bait situation.
6. the production method of a kind of cross-sterility spotted maigre according to claim 1, it is characterised in that：Step（7）Inspection
In the success rate of cross hybrid seedling infertility, when cross hybrid seedling was cultivated to 3 monthly age, 30 tails are randomly selected, sexual gland is taken after dissection, sexual gland is presented
It is thin-line-shaped；Sexual gland is fixed to be changed after 12h using Bonn formula liquid and is preserved using 70% alcohol；After the dehydration of sexual gland sample, FFPE
Histotomy is carried out, the Gonadal Morphology after section is without spermary lobule or ovarian structure, therefore it is completely sterile to be accredited as sexual gland.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107821244A (en) * | 2017-11-16 | 2018-03-23 | 浙江省海洋水产研究所 | A kind of ratio of female high little yellow croaker breeding method and feedstuff feeding device |
| CN107821244B (en) * | 2017-11-16 | 2019-09-24 | 浙江省海洋水产研究所 | A kind of little yellow croaker breeding method and feedstuff feeding device that ratio of female is high |
| CN108812447A (en) * | 2018-05-09 | 2018-11-16 | 浙江省海洋水产研究所 | It is a kind of to hybridize reproduction cell transplantation method of the spotted maigre adult fish as receptor |
| WO2020033940A1 (en) * | 2018-08-10 | 2020-02-13 | Center For Aquaculture Technologies, Inc. | A method of generating sterile and monosex progeny |
| CN113163740A (en) * | 2018-08-10 | 2021-07-23 | 水产养殖技术中心有限公司 | Method for producing sterile and parthenogenetic progeny |
| CN113163740B (en) * | 2018-08-10 | 2023-09-08 | 水产养殖技术中心有限公司 | Method for producing sterile and unisexual offspring |
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| CN106614142B (en) | 2019-12-03 |
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