CN110463664A - A kind of preparation method of Hucho taimen triploid seed - Google Patents
A kind of preparation method of Hucho taimen triploid seed Download PDFInfo
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- CN110463664A CN110463664A CN201910832172.8A CN201910832172A CN110463664A CN 110463664 A CN110463664 A CN 110463664A CN 201910832172 A CN201910832172 A CN 201910832172A CN 110463664 A CN110463664 A CN 110463664A
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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
- 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; CARE OF BIRDS, FISHES, INSECTS; 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
- A01K61/17—Hatching, e.g. incubators
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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
- A01K67/00—Rearing or breeding animals, not otherwise provided for; New breeds of animals
- A01K67/027—New breeds of vertebrates
- A01K67/0275—Genetically modified vertebrates, e.g. transgenic
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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
- 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
Abstract
The present invention provides a kind of preparation method of Hucho taimen triploid seed, including triploid heat shock induction, and in diploid fertilized eggs after fertilization 15min, which is put into transition 1min in 17 DEG C of water-baths；It is then placed in 26-27 DEG C of water-bath and is heat-treated 15min；Place into transition 1min in 17 DEG C of water-baths；Fertilized eggs through the triploid heat shock induction are placed in the container of 7-9 DEG C of hatching water temperature and hatch by incubating oosperm.Hucho taimen triploid prelarva can be obtained within this method 30-40 days, fertilized eggs hair eye rate is 80% or more during hatching, and 70% or more hatching rate, triploid rate reaches 90% or more.The method of the present invention was effectively relieved due to the problems such as speed of growth reduces, coefficient of condition reduces, the death rate is high after parent population breeding after sexal maturity, premunition is improved, suitable for manufacturing the big specification fish of raw fish, is had at low cost, the advantages that easy to operate, has preferable market value.
Description
Technical field
The present invention relates to Hucho taimen breeding technical fields, and in particular to a kind of preparation method of Hucho taimen triploid seed.
Background technique
Hucho taimen (Hucho taimen) also known as taimen belong to salmon shape mesh, and salmonidae, taimen category is that the world is rare in imminent danger
Cold water fishes is mainly distributed on Northern Asia area, is distributed mainly on the Heilungkiang, the Yalu River, Tumen of northeast in China
The waters such as the Eerqisihe River in river and Xinjiang, Wild population quantity is extremely rare at present, has important protection and economic value.
Hucho taimen parent population has the habit buried ovum and protect nest, and parent population mortality after oviposition is especially more with milter.Since disposition is violent,
Male and female sex character is unobvious, the more difficult grasp of artificial propagation techniques.Big specification triploid Hucho taimen is cultivated, flesh of fish ratio can be improved
Rate is suitable for processing raw fish, and alleviates the problem of parent population postpartum death after gonadal maturation.Hucho taimen triploid induction skill
There is not been reported both at home and abroad for art.
Summary of the invention
Since the death rate of Hucho taimen fertilized eggs for a long time is higher, and it is difficult to improve hair eye rate, hatching rate and triploid
Rate.On the basis of lot of experiments, the present inventor is found surprisingly that, in optimization Hucho taimen diploid fertilized eggs
In the case where heat treatment time and temperature, the fertilized eggs after thermally treated 15min are further placed into transition in 17 DEG C of water-baths
1min can reduce significantly the death rate of fertilized eggs, and also improve hair eye rate, hatching rate and triploid rate.
Specifically, the present invention provides a kind of preparation method of Hucho taimen triploid seed, comprising:
The step of triploid heat shock induction, which is included in diploid fertilized eggs after fertilization 15min, by the fertilized eggs
It is put into transition 1min in 17 DEG C of water-baths；It is then placed in 26-27 DEG C of water-bath and is heat-treated 15min；Place into transition in 17 DEG C of water-baths
1min；And
The step of incubating oosperm, the step include that the fertilized eggs through the triploid heat shock induction are placed in 7-9 DEG C
Hatch and is hatched in the container of water temperature.
Preferably, the step of triploid heat shock induction, is included in diploid fertilized eggs after fertilization 15min, by this by
Smart ovum is put into transition 1min in 17 DEG C of water-baths；It is then placed in 26.5 DEG C of water-baths and is heat-treated 15min；Place into mistake in 17 DEG C of water-baths
Cross 1min.Experiment discovery, using the abductive approach can make the fertilized eggs 2h death rate drop to 3.6% hereinafter, hair eye rate improve to
81%, hatching rate is improved to 72%, and triploid rate is improved to 91%, and comprehensive score is up to 85 points.
Specifically, details are as follows for the step of triploid heat shock induction:
After fertilization 15min hangs the stainless steel that fertilized eggs are put into diameter 15cm in leakage, first in 17 DEG C of water temperature of water-bath
Transition 1min is placed into the electronic thermostatic sink of 26-27 DEG C (preferably 26.5 DEG C), is gently teetertottered and slip through the net, make fertilized eggs
It is consistent with water temperature around as early as possible.It is heat-treated the first transition 1min in 17 DEG C of water temperature of water-bath of fertilized eggs after 15min, then puts
Enter and stood in the container of 7-9 DEG C of hatching water temperature, 2h detects dead ovum, counts the death rate, places into the container of 7-9 DEG C of hatching water temperature
In, into normal incubation.It is that the closed kitchen utensils brine of upper cover is blue that stainless steel, which hangs leakage, and surrounding is uniformly distributed filling for diameter 2mm
Hole, it is desirable that inner wall smooth.Water-bath is laboratory conventional heating container, 1-99 DEG C of temperature control range, Cha≤0.5 DEG C Wu, sensitivity
0.2℃.Electronic thermostatic sink is equipped with circulating pump, makes water body flow, controls 5-99 DEG C of temperature range, accidentally poor≤0.5 DEG C, sensitivity
0.1℃.With high-precision temperature meter calibrating when temperature is set.
When water temperature is lower than 26.5 DEG C, the processing time less than in the case where 20min, fertilized eggs heat shock before and after the processing, 17
Transition 1min in DEG C water, can reduce stimulation of the excessive temperature differentials to ovum, the 2h death rate can control below 5%.After fertilization
15min is heat-treated 15min in 26.5 DEG C of water-baths, and triploid rate is up to 91%.
Further, the method for incubating oosperm of hatching barrel+place side by side slot combination can be used in the present invention, and fertilized eggs hair is used at the moment
Hatching barrel is hatched, and is moved into after hair eye and is placed side by side slot.
Specifically, details are as follows for method for incubating oosperm:
Rainbow trout conventional hatching methods can be used.Fertilized eggs hair is hatched with hatching barrel at the moment, is moved into after hair eye and is placed side by side slot.It incubates
Change bucket is commercially available non-toxic plastic bucket, and interior arrangement is referring to rainbow trout hatching barrel, 4-5 ten thousand, specification 10L barreled ovum.Slot is placed side by side as food
The production of grade resin material, outside large trough specification are 300cm × 45cm × 27cm, internal mating 5 sulculuses, small trench bottom and are gone out
Water side is aperture 2mm microwell plate, and specific structure is hatched referring to rainbow trout.Incubation period controls 7-10 DEG C of water temperature, dissolved oxygen >
6mg/L.Hair sterilizes 1h with 50ppm formaldehyde flowing water weekly at the moment.Entire incubation period accumulated temperature needs 260-280 to subsist, the incubation period
About 30-40 days.
Incubation period adjusts water temperature at 7-10 DEG C with water storage box, avoids water temperature from floating excessive.Periodically use formaldehyde drug disinfection, control
Water mo(u)ld processed breeds, and hatching rate is up to 70% or more.
Further, the preparation method of Hucho taimen triploid seed provided by the invention further includes rejecting in after fertilization 2h
Dead ovum, is then hatched again.Since heat treatment has certain injury to ovum, the 2h death rate is to examine the important finger of inducing temperature
Mark.In addition, can influence the hatching of normal ovum if not graning except dead ovum is directly hatched because dead ovum breeds water mold, reduce hatching rate.
Further, the preparation method of Hucho taimen triploid seed provided by the invention, further includes the steps that parent fish rearing.
The step can promote parent population property under artificial condition by establishing the genital regulating system based on fortification and thermal stimulus
Gland is reached maturity.
Specifically, details are as follows for the step of parent fish rearing:
The parent fish live through the winter previous season of growth just carries out parent fish selection and cultivation.It cultivates water temperature to require at 14~17 DEG C, water
Middle dissolved oxygen is required in 6mg/L or more, feeds salmon trout parent population special feed (protein in feed by 1.5% feeding rate daily
Content should be higher than that 40%, and 10 kinds of essential amino acids should keep balancing), a fresh and alive fish is fed by 5%~8% feeding rate within every 10 days
Bait.Winter, every two days of overwintering period fed parent population by 0.3% feeding rate overwintering at least 60 days or more under 2-4 DEG C of water temperature condition
Feed feeds primary fresh and alive bait by 5% feeding rate in every 10 days.Spring water temperature is raised to 6~7 DEG C, and oxygen in water requires
8mg/L or more feeds parent population feed by 0.5% feeding rate daily, feeds primary fresh and alive bait by 3%~5% feeding rate daily,
Reinforced cultivating before being laid eggs can be carried out artificial propagation in 20 days or so.To promote yield in gonad development and domination set,
It is blent with the underground water of different temperatures or mountain spring water, adjusts water temperature.Parent fish rearing need to carry out under discharge condition.Parent population year
Age: raun 6+It is more than age, milter 5+It is more than age；Parent population specification: male parent population 3000g or more, female parent population 3500g or more.Female milter number
Amount proportion 1:1.
Further, the preparation method of Hucho taimen triploid seed provided by the invention further includes obtaining diploid fertilization
The step of ovum.The present invention hastens parturition the parent population of gonadal maturation through injecting artificial hormone, and dry method fertilization obtains common diploid
Fertilized eggs.
Since Hucho taimen parent population is there may be the nonsynchronous phenomenon of gonad development, the single drug that conventional method is taken is hastened parturition
Effect is undesirable.To solve this problem, the present invention adjusts induced spawning method, is changed to hasten parturition using drug twice, by low for the first time
Accelerating the ripening for dosage makes male and female parent population gonad development tend to be synchronous.
In addition, should not be controlled during artificial induced spawning since Hucho taimen parent population figure is larger.The present invention is used when adopting ovum
0.03% ethylene glycol phenyl ether anaesthetizes parent population.Compared with conventional method, the method for the present invention reduces parent population in crowded ovum process
In stress reaction, not only increase parent population postnatal survival rate, and reduce manually crowded damage of the ovum to ovum.
Unlike conventional method essence ovum hybrid manipulation, the present invention filter adopt ovum generation coelomic fluid (such as with leak
Net), and further control sex ration is 1:3 fertilization, can improve rate of fertilization significantly in this way.
Continue the vigor time to further extend sperm, the present invention has also prepared 0.6% NaCl dilution, is inseminating
Hucho taimen sperm is placed in the dilution at room temperature temporarily preservation in the process, sperm energy can be reduced in such short time
Loss, will not influence its vigor, this is very necessary for improving rate of fertilization.
Specifically, details are as follows for the step of acquisition diploid fertilized eggs:
Artificial induced spawning, ocyodinic are No. 2 (LHRH-A of fish ovulation induction element2) and maleic acid dioxone (DOM).Using pectoral fin
2 injection methods of base portion, the 1st injection time are antenatal 1 month, 6 DEG C of water temperature, raun LH injection RH-A2, dosage 1ug/kg,
To dematuration；2nd injection, raun LHRH-A25ug/kg+DOM 5mg/kg；Milter dosage halves.Hormone dosage view property
Gland development degree and water temperature appropriate adjustment.Effector phase 7-10 days.
With 0.03% ethylene glycol phenyl ehterization when adopting ovum, parent population stress reaction is reduced.It adopts and filters coelomic fluid with slipping through the net after ovum,
The female fish-egg of 1 tail of the time of fertilization is fertilized with 3 tail milter sperm dry method；The NaCl dilution that 0.6% is prepared in Process of Insemination, by wise sieve
Salmon sperm temporarily saves at room temperature, to extend sperm life.Adopt 7-9 DEG C of ovum water temperature.Smart ovum feather stirring 15-20s makes
Smart ovum is uniformly mixed, and a small amount of clear water is added and is slowly stirred 1min, then is rinsed 2-3 times with clear water, washes away excessive sperm, ovum skin, blood
Block is added clear water and places, to ovum water swelling.
It adopts ovum and fertilization process avoids direct sunlight.Through stimulation by running water, fortification, drug is hastened parturition twice, adopts ovum water temperature 6
DEG C or more, spawning rate is up to 90% or more.
The present invention adopts ovum opportunity by grasping, and prevents ovum overdone, and male and female 1:3 fertilization slips through the net and filters coelomic fluid, prepares
0.6% NaCl dilution is used as Semen routine, and rate of fertilization is up to 80% or more.With 0.03% ethylene glycol phenyl ehterization when adopting ovum
Parent population stress reaction, parent population postpartum survival rate 100% can be reduced.
Pactec CyFlow Cube8 type flow cytomery fry blood DNA relative amount can be used in the present invention
Method determines triploid rate, to carry out Ploidy detection.
Specifically, details are as follows for the ploidy detection method:
It is determined using the method for Pactec CyFlow Cube8 type flow cytomery fry blood DNA relative amount
Triploid fish.Fry rearing to 5-8cm, every group of 50 tail of stochastical sampling of test group, tail vein living body blood drawing detects 3 tails two every time
Times body fry is as control.
Hucho taimen triploid prelarva can be obtained within the method for the present invention 30-40 days, fertilized eggs hair eye rate is reachable during hatching
80% or more, hatching rate is up to 70% or more, and for obtained prelarva through flow cytomery, triploid rate reaches 90% or more.
After the method for the present invention can be effectively relieved since the speed of growth reduces, coefficient of condition reduces, parent population breeds after sexal maturity
The problems such as death rate is high improves premunition, suitable for manufacturing the big specification fish of raw fish, have it is at low cost, it is easy to operate etc.
Advantage has preferable market value.
Specific embodiment
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..It is not specified in embodiment specific
Technology or conditions person, described technology or conditions according to the literature in the art, or carried out according to product description.It is used
Production firm person is not specified in reagent or instrument, is the conventional products that can be commercially available by regular distributor.
Embodiment 1
A kind of preparation method of Hucho taimen triploid seed, specifically comprises the following steps:
1,6 parent fish rearing: are selected+More than, physical strong, well-developed parent population artificial culture, male parent population specification 3000g
More than, female parent population specification 3500g or more.Female milter proportion is 1:1.Feed is salmon trout parent population special compound feed, and albumen contains
40% or more amount, periodically feeds fresh and alive bait.It need to strengthen cultivation within antenatal 1 month.
2, the acquisition of diploid fertilized eggs: 6 DEG C of minimum water temperature of hastening parturition, ocyodinic are the LHRH- of Ningbo hormone head factory production
A2And DOM.Antenatal every kilogram of fish LHRH-A of 1 month raun21ug accelerates the ripening.Production the last week hastens parturition dosage as every kilogram of fish 5ug of raun
LHRH-A2+5mg DOM；Milter halves.Effector phase 7-10 days.With 0.03% ethylene glycol phenyl ehterization parent population when adopting ovum.After adopting ovum
Coelomic fluid is filtered with slipping through the net, male and female 1:3 dry method fertilization, the NaCl dilution for preparing 0.6% is used as Semen routine.Adopt ovum water temperature 7-
9℃.Smart ovum feather stirring 15-20s is uniformly mixed smart ovum, and a small amount of clear water is added and is slowly stirred 1min, then is rinsed with clear water
2-3 times, excessive sperm, ovum skin, clot are washed away, clear water is added and places, to ovum water swelling.
3, triploid heat shock induction: fertilized eggs are put into diameter 15cm surrounding envelope by diploid fertilized eggs after fertilization 15min
The stainless steel closed is hung in leakage, and first the transition 1min in 17 DEG C of water temperature of water-bath, places into 26.5 DEG C of electronic thermostatic sink,
It gently teetertotters and slips through the net, keep fertilized eggs consistent with water temperature around as early as possible.Fertilized eggs are put into water temperature again after heat treatment 15min
Then transition 1min in 17 DEG C of water-bath is put into 7-9 DEG C of hatching water and stands, after fertilization 2h detects dead ovum and counts, and directly puts
In the container for entering 7-9 DEG C of hatching water temperature, into normal incubation.
This link operational motion is light.
3, it incubating oosperm: carries out indoors, it is desirable that it is rather dark, it avoids shaking.Fertilized eggs hair is incubated with hatching barrel at the moment
Change, general 10L barrels puts 4~50,000 fertilized eggs, and for every barrel of water injection rate in 3~6L/min, accumulated temperature 170, which is subsisted, sends out eye, and hair eye moves back
Enter to placing side by side slot.100 ovum of grab sample after eye are sent out, it is eyed eggs that idiosome, which has obvious black eyespot, detects non-eyed eggs, white
Ovum calculates hair eye rate.Membrane phase, statistics detect dead ovum number, calculate hatching rate.7-10 DEG C of incubation period water temperature.Dissolved oxygen > 5mg/
L.Entire incubation period accumulated temperature needs 260-280 to subsist；From rupture of membranes to all about 7-10 days time of hatching.The result shows that 2h
The death rate 3.6%, hair eye rate 81%, hatching rate 72%, triploid rate 91%, comprehensive score 85 are divided.
Ploidy detection method used by the present embodiment: Pactec CyFlow Cube8 type flow cytomery is used
The method of fry blood DNA relative amount determines triploid rate.Fry rearing is to 5-8cm, every group of 50 tail of stochastical sampling of test group,
The blood drawing of tail vein living body detects 3 tail diploid fish fries as control every time.
Experiment 1
Difference with embodiment 1 is only that, changes the step 3 fertilized eggs heat shock induction initiated process times, heat treatment temperature
The time is spent and is persistently handled, 11 groups of experiments are set, each group step 3 processing method and related experiment result are as follows:
1) after fertilization 10min, 26 DEG C of processing 10min, the 2h death rate 0.5%, hair eye rate 84%, hatching rate 79%, three times
Body rate 19%, comprehensive score 38 are divided；
2) after fertilization 15min, 26 DEG C of processing 15min, the 2h death rate 0.13%, hair eye rate 82%, hatching rate 76%, three times
Body rate 75%, comprehensive score 79.2 are divided；
3) after fertilization 20min, 26 DEG C of processing 20min, the 2h death rate 1.24%, hair eye rate 58%, hatching rate 59%, three times
Body rate 66%, comprehensive score 61.2 are divided；
4) after fertilization 10min, 26.5 DEG C of processing 10min, the 2h death rate 0.89%, hair eye rate 86%, hatching rate 77%, three
Times body rate 42%, comprehensive score 68.4 are divided；
5) after fertilization 20min, 26.5 DEG C of processing 20min, the 2h death rate 14.68%, hair eye rate 68%, hatching rate 61%,
Triploid rate 60%, comprehensive score 64.8 are divided；
6) after fertilization 10min, 27 DEG C of processing 10min, the 2h death rate 5.7%, hair eye rate 66%, hatching rate 65%, three times
Body rate 47%, comprehensive score 50.4 are divided；
7) after fertilization 15min, 27 DEG C of processing 15min, the 2h death rate 8.8%, hair eye rate 68%, hatching rate 44%, three times
Body rate 88%, comprehensive score 76 are divided；
8) after fertilization 20min, 27 DEG C of processing 20min, the 2h death rate 13.39%, hair eye rate 37%, hatching rate 48%, three
Times body rate 67%, comprehensive score 49 are divided；
9) after fertilization 10min, 28 DEG C of processing 10min, the 2h death rate 16.41% send out eye rate 48.4%, hatching rate
28.6%, triploid rate 49%, comprehensive score 48.6 is divided；
10) after fertilization 15min, 28 DEG C of processing 15min, the 2h death rate 21.95% send out eye rate 46.4%, hatching rate
25.9%, triploid rate 85%, comprehensive score 61.8 is divided；
11) after fertilization 20min, 28 DEG C of processing 20min, the 2h death rate 24.39%, hair eye rate 0.65%, hatching rate 9%,
Triploid rate 52%, comprehensive score 21.2 are divided.
Experiment 2
Difference with embodiment 1 is only that omitting in step 3 " will be put into water temperature 17 for fertilized eggs again after heat treatment 15min
DEG C water-bath in transition 1min " the step of, i.e., by 26.5 DEG C of electronic thermostatic sink be heat-treated 15min after fertilized eggs
It being placed directly in 7-9 DEG C of hatching water and stands, after fertilization 2h detects dead ovum and counts, it is directly placed into the container of 7-9 DEG C of hatching water temperature,
Into normal incubation.
The fertilized eggs heat shock induction initiated process time of three groups of processing, heat treatment temperature persistently handle the time below, and
Related experiment result is as follows:
1) after fertilization 15min, 26 DEG C of processing 15min, (transitional link after lacking the heat treatment), the 2h death rate 6.9%,
Hair eye rate 62%, hatching rate 66%, triploid rate 77%, comprehensive score 67 are divided；
2) after fertilization 15min, 26.5 DEG C of processing 15min, (transitional link after lacking the heat treatment), the 2h death rate
9.6%, eye rate 59% is sent out, hatching rate 62%, triploid rate 73%, comprehensive score 64.6 divides；
3) after fertilization 15min, 27 DEG C of processing 15min, (transitional link after lacking the heat treatment), the 2h death rate
13.4%, eye rate 48% is sent out, hatching rate 54%, triploid rate 68%, comprehensive score 56 divides.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, fall within the scope of the claimed invention without departing from theon the basis of the spirit of the present invention.
Claims (8)
1. a kind of preparation method of Hucho taimen triploid seed characterized by comprising
The step of triploid heat shock induction, it is included in diploid fertilized eggs after fertilization 15min, which is put into 17 DEG C of water
Transition 1min in bath；It is then placed in 26-27 DEG C of water-bath and is heat-treated 15min；Place into transition 1min in 17 DEG C of water-baths；And
The step of incubating oosperm, including the fertilized eggs through the triploid heat shock induction are placed in 7-9 DEG C of hatching water temperature
Hatched in container.
2. preparation method according to claim 1, which is characterized in that the step of triploid heat shock induction is included in
The fertilized eggs are put into transition 1min in 17 DEG C of water-baths by diploid fertilized eggs after fertilization 15min；It is then placed in 26.5 DEG C of water-baths
It is heat-treated 15min；Place into transition 1min in 17 DEG C of water-baths；Alternatively,
The step of triploid heat shock induction, is included in diploid fertilized eggs after fertilization 15min, which is put into 17
Transition 1min in DEG C water-bath；It is then placed in 26 DEG C of water-baths and is heat-treated 15min；Place into transition 1min in 17 DEG C of water-baths；Alternatively,
The step of triploid heat shock induction, is included in diploid fertilized eggs after fertilization 15min, which is put into 17
Transition 1min in DEG C water-bath；It is then placed in 27 DEG C of water-baths and is heat-treated 15min；Place into transition 1min in 17 DEG C of water-baths.
3. preparation method according to claim 1 or 2, which is characterized in that further include rejecting dead ovum in after fertilization 2h, then
Hatched again.
4. preparation method according to claim 1-3, which is characterized in that further include the steps that parent fish rearing；Its
In, the water temperature of the cultivation is required at 14~17 DEG C, and oxygen in water requires to throw by 1.5% feeding rate daily in 6mg/L or more
Salmon trout parent population special feed is fed, feeds primary fresh and alive bait by 5%~8% feeding rate within every 10 days；Winter is in 2-4 DEG C of water temperature item
Overwintering at least 60 days or more under part, every two days of overwintering period fed parent population feed by 0.3% feeding rate, pressed 5% bait throwing within every 10 days
Rate feeds primary fresh and alive bait；Spring water temperature is raised to 6~7 DEG C, and oxygen in water requires to throw by 0.5% daily in 8mg/L or more
Bait rate feeds parent population feed, feeds primary fresh and alive bait by 3%~5% feeding rate daily, the reinforced cultivating before being laid eggs；
And/or
The parent population age: raun 6+It is more than age, milter 5+It is more than age；The parent population specification: male parent population 3000g or more, female parent population
3500g or more；Female milter quantitative proportion 1:1.
5. preparation method according to claim 1-4, which is characterized in that further include obtaining diploid fertilized eggs
Step；
Wherein, ocyodinic used in artificial induced spawning is No. 2 (LHRH-A of fish ovulation induction element2) and maleic acid dioxone (DOM), it uses
2 injection methods of pectoral fin base portion, the 1st injection time are antenatal 1 month, 6 DEG C of water temperature, raun LH injection RH-A2, dosage 1ug/
kg；2nd injection, raun LHRH-A25ug/kg+DOM 5mg/kg；Milter dosage halves；And/or
With 0.03% ethylene glycol phenyl ehterization parent population to reduce stress reaction when adopting ovum；And/or
It adopts and filters coelomic fluid with slipping through the net after ovum, the female fish-egg of 1 tail of the time of fertilization is fertilized with 3 tail milter sperm dry method.
6. preparation method according to claim 5, which is characterized in that in the acquisition diploid fertilized eggs the step of also
Including Hucho taimen sperm is placed in 0.6%NaCl dilution at room temperature temporarily preservation.
7. preparation method according to claim 1-6, which is characterized in that the step of incubating oosperm uses
Hatching barrel+place side by side the method for incubating oosperm that slot combines, fertilized eggs hair is hatched with hatching barrel at the moment, is moved into and is placed side by side after hair eye
Slot.
8. preparation method according to claim 1-7, which is characterized in that incubated in the step of the incubating oosperm
The change phase controls 7-10 DEG C of water temperature, dissolved oxygen > 6mg/L；And/or
Hair sterilizes 1h with 50ppm formaldehyde flowing water weekly at the moment；And/or
Entire incubation period accumulated temperature needs 260-280 to subsist, and the incubation period about 30-40 days.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112877445A (en) * | 2021-03-08 | 2021-06-01 | 河南牧业经济学院 | Microsatellite marker and primer for identifying genetic sex of hucho taimen, and identification method and application |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4697546A (en) * | 1986-05-30 | 1987-10-06 | Purdue Research Foundation | Method for production of tetraploid channel catfish |
| JPH10150883A (en) * | 1996-11-21 | 1998-06-09 | Kanagawa Pref Gov | Method for creating total male triploid fish by using sex reversal male fish of ayu |
| CN102696515A (en) * | 2012-06-01 | 2012-10-03 | 北京市水产科学研究所 | Preparation method of game fish triploid fries |
| CN105123566A (en) * | 2015-07-23 | 2015-12-09 | 新疆农业大学 | Hucho taimen sperm activation solution and preparation method thereof |
| CN107810888A (en) * | 2017-12-01 | 2018-03-20 | 北京市水生野生动植物救护中心 | A kind of method for culturing seedlings of fine-scaled graphite triploid seed |
-
2019
- 2019-09-04 CN CN201910832172.8A patent/CN110463664A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4697546A (en) * | 1986-05-30 | 1987-10-06 | Purdue Research Foundation | Method for production of tetraploid channel catfish |
| JPH10150883A (en) * | 1996-11-21 | 1998-06-09 | Kanagawa Pref Gov | Method for creating total male triploid fish by using sex reversal male fish of ayu |
| CN102696515A (en) * | 2012-06-01 | 2012-10-03 | 北京市水产科学研究所 | Preparation method of game fish triploid fries |
| CN105123566A (en) * | 2015-07-23 | 2015-12-09 | 新疆农业大学 | Hucho taimen sperm activation solution and preparation method thereof |
| CN107810888A (en) * | 2017-12-01 | 2018-03-20 | 北京市水生野生动植物救护中心 | A kind of method for culturing seedlings of fine-scaled graphite triploid seed |
Non-Patent Citations (3)
| Title |
|---|
| 上海水产学院: "《组织胚胎学》", 31 May 1981, 农业出版社 * |
| 李道友: "《名优水产品高校养殖技术》", 31 December 2009, 贵州科学技术出版社 * |
| 陈春山等: "图们江细鳞鲑人工繁殖技术研究", 《水生态学杂质》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112877445A (en) * | 2021-03-08 | 2021-06-01 | 河南牧业经济学院 | Microsatellite marker and primer for identifying genetic sex of hucho taimen, and identification method and application |
| CN112877445B (en) * | 2021-03-08 | 2023-10-24 | 河南牧业经济学院 | Microsatellite marker and primer for genetic sex identification of hucho taimen as well as identification method and application |
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