CN109392775A - A kind of selection being sheerly all-male hybridization Pelteobagrus fulvidraco and its parent - Google Patents
A kind of selection being sheerly all-male hybridization Pelteobagrus fulvidraco and its parent Download PDFInfo
- Publication number
- CN109392775A CN109392775A CN201811410043.1A CN201811410043A CN109392775A CN 109392775 A CN109392775 A CN 109392775A CN 201811410043 A CN201811410043 A CN 201811410043A CN 109392775 A CN109392775 A CN 109392775A
- Authority
- CN
- China
- Prior art keywords
- pelteobagrus
- raun
- male
- fish
- pure lines
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Withdrawn
Links
Classifications
-
- 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
-
- 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 kind of pure lines all-male hybridization Pelteobagrus fulvidraco and its selections of parent, the wild original excellent from the selected character of Dongting Lake System is Pelteobagrus fulvidraco and Pelteobagrus vachelli, synchronous applications cell engineering Biotechnology in Genetic Breeding, it is variety rejuvenation to wild original, selects pure lines Pelteobagrus fulvidraco and the pure lines full raun of Pelteobagrus vachelli；To be sheerly the full raun of Wa Shi as raw material fish, high-quality Pelteobagrus vachelli supermale fish is researched and developed；And establish the full raun of pure lines and supermale fish Germplasm Bank.It constantly purificates and rejuvenates to original seed, it is ensured that seedling is pure, gene are stably and controllable.High-quality Pelteobagrus vachelli supermale fish combines with the pure lines full raun of Pelteobagrus fulvidraco, and extensive artificial propagation pure lines all-male hybridizes advanced fry of yellow catfish.The present invention thoroughly solves the problems such as hybridization Pelteobagrus fulvidraco germplasm is degenerated from generation to generation, growth vigor declines, disease-resistant resistance against disadvantage is weak from parent source and cultivates overall process not medication substantially, it is ensured that food safety since seed.
Description
Technical field
The invention belongs to fish cell engineering Biotechnology in Genetic Breeding fields, and in particular to a kind of pure lines all-male hybridization Pelteobagrus fulvidraco
And its selection of parent.
Background technique
Crossbreeding technology is widely adopted because hybrid seedlings have heterosis, hybrid vigor.But hybridize the choosing of Pelteobagrus fulvidraco Parent
Educate with purification and rejuvenation work, ignored by people always, sieve bottom " mao mao fish " plus commodity in use fish year after year and make parent's inbreeding,
Germplasm decreases sharply, and fish disease takes place frequently, and medication aggravates, and endangers food safety.Though there are many traditional select index methods, effect is best
Person is continuous 14 generation Sibling's inbreeding breeding variety, but this method needs time-consuming 14-20 years, and intermediate link slightly has
Mistake, then all that has been achieved is spoiled, it is necessary to which accent is done.And use high-tech means --- apply fish cell engineering genetic breeding skill
Art is that parent carries out two generation gynogenesis to wild original, enables its gene loci completely homozygous, only last 2 years and just reach continuous ten
The purity of the Sibling's inbreeding of four generations (and period operates none fault of necessary overall process) breeding, that is, reach pure lines original seed
Criterion.In addition, since hybridization Pelteobagrus fulvidraco male and female individual and long speed difference are different up to 2-3 times, it is necessary to holandric inheritance breeding technique is applied,
Producing genetic pure lines all-male hybridization advanced fry of yellow catfish replaces standard hybridization seedling to cultivate for the whole nation.
Summary of the invention
For overcome the deficiencies in the prior art, the present invention provides a kind of breeding for being sheerly all-male hybridization Pelteobagrus fulvidraco and its parent
Method.
The technical solution adopted in the present invention is mainly: a kind of breeding side being sheerly all-male hybridization Pelteobagrus fulvidraco and its parent
Method, comprising the following steps:
(1) the pure lines full raun Germplasm Bank of Pelteobagrus fulvidraco is established, is Pelteobagrus fulvidraco raun by two generation gynogenesis, choosing by wild original
The complete female Pelteobagrus fulvidraco of pure lines is brought out, and establishes the pure lines full raun Germplasm Bank of Pelteobagrus fulvidraco, wherein first generation gynogenesis includes following step
It is rapid:
A, the preparation of the Heterologous Sperm of inhereditary material inactivation, the southern catfish milter for being derived from Pearl River system is infused by kg body weight
It penetrates LHRH-A2 5-10 μ g to accelerate the ripening, smart 2g is manually taken after 10 hours, dilute 3 times with spermatozoa preservative fluid, stir evenly and be laid in glass
On vessel, thickness is no more than 0.5mm, is placed on microoscillator, starts microoscillator, with 2537 angstroms of ultraviolet radiator of wavelength
Make radiation source, 25min is irradiated, to obtain the Heterologous Sperm of genetic inactivation；
B, the collection and insemination of fish-egg will be Pelteobagrus fulvidraco from " 800 inner Dongting Lake " lake region wild original chosen on fishing boat of fishing
Raun presses per kilogram of body weight LH injection RH-A2 12 μ g, DOM 5mg, HCG 600IU, physiological saline 5ml；Parent population temporarily supports water temperature 28
DEG C, fish belly is squeezed after 12-15 hours, collects fish-egg；It extracts above-mentioned Heterologous Sperm to be added in fish-egg, mixes, add appropriate essence
Sub- activating fluid, stirs evenly, and stands insemination work in 2 minutes and completes；
C, the induction of gynogenetic diploid, when above-mentioned artificial insemination, the Heterologous Sperm of genetic inactivation is not involved in the hair of fertilized eggs
It educates, only raun fish-egg is developed and Haploid production, and monoploid seedling can not survive, for this purpose, by cold shock method, resistance
Disconnected meiosis second polar body is discharged and induces gynogenetic diploid, since the sexual control mechanism of Pelteobagrus fulvidraco is that male gamete is different
Type, female gamete homotype, so ♀ XX Gynogenetic Offspring is complete female filial generation；
Second generation gynogenesis: first generation Gynogenetic Offspring is cultivated to sexually matured raun, is carried out according to the above method
Second generation gynogenesis can obtain the completely homozygous pure lines Pelteobagrus fulvidraco female filial generation entirely of gene loci；
It is sheerly the foundation of the full raun Germplasm Bank of Pelteobagrus fulvidraco: the full raun ♀ of pure lines Pelteobagrus fulvidraco that second generation gynogenesis obtains
XX, opening take out wherein 500 tails and carry out artificial turning property, obtain the physiology milter ♂ XX with normal reproduction ability after 6 days, will be pure
It is that full raun ♀ XX and physiology milter ♂ XX are cultivated to sexal maturity, hybridizes group expanding, establishes the pure lines full raun Germplasm Bank of Pelteobagrus fulvidraco,
Mass production is sheerly the full raun of Pelteobagrus fulvidraco and makees female parent for large-scale breeding pure lines all-male hybridization Pelteobagrus fulvidraco；
(2) the pure lines full raun Germplasm Bank of Pelteobagrus vachelli is established, breeding is sheerly the full raun ♀ XX of Pelteobagrus vachelli, and establishes
It is sheerly the full raun Germplasm Bank of Wa Shi, operating process is identical as the above-mentioned full raun of pure lines Pelteobagrus fulvidraco, is only parent population by wild original
It is that be changed to wild original be Pelteobagrus vachelli raun to Pelteobagrus fulvidraco raun；
(3) research and development of high-quality Pelteobagrus vachelli supermale fish and its foundation of Germplasm Bank, comprising the following steps:
A, the well-developed full raun ♀ XX of pure lines Pelteobagrus vachelli is chosen in the production of hybrid seeds of high-quality Pelteobagrus vachelli physiology raun
The wild original particularly excellent with germplasm is that Pelteobagrus vachelli milter ♂ XY mates breeding, hatches high-quality Pelteobagrus vachelli filial generation ♀
XX and ♂ XY, fry are open the 6th day, take out 1000 tail Pelteobagrus vachelli sprays and independently cultivate, with 17-that concentration is 150 μ g/L
α ethinyl estradiol impregnate after fairy shrimp live bait carry out feeding, one day 3 eat, even feed 50 days, later with conventional feed feed to
Sexal maturity, this write instructions and transfer property Pelteobagrus vachelli in, physiology raun and common raun respectively account for 50%；
B, test cross selects high-quality Pelteobagrus vachelli supermale fish ♂ YY and supermale physiology raun ♀ YY, from above-mentioned direct cultivation
In mature high-quality Pelteobagrus vachelli, 2-3 tails individual maximum, the milter that external physical characteristic is most excellent, gonad development is most mature are chosen
♂ XY, dissection are taken out spermary, are ground, and spermatozoa preservative fluid is added to be made into the dilution sperm of 1:3, sealing be put into 4 DEG C of refrigerators save it is standby
With；In addition from being cultivated in mature high-quality Pelteobagrus vachelli raun after above-mentioned turning property, 20 tails individual maximum, resemblance are chosen most
The most mature pseudo- raun of outstanding, gonad development, squeezes out fish-egg respectively after hastening parturition, independently carry out with above-mentioned ready sperm artificial
Insemination, fish-egg is uniformly layered in mesh sheet and is independently hatched；After fry hatching membrane, the fry that each tail mother fish produces all is cooked easy to remember
Record, it is independent to cultivate, it cannot mix.It is open after the 6th day, the spray that this 20 tail parent population produces respectively is selected and remain into 200 tails, and be divided into A, B two
Group, every group of 100 tails, A group are directly fed with conventional feed, and group two continues to raise according to a conventional method after being feminized with pharmaceutical chemistry；3 months
Afterwards, the gender of A group is determined, if half male and half female, the maternal sex chromosomal gene of the group is ♀ XX, then all by two groups of A, B
It eliminates；If in A group filial generation, male accounts for 75%, and female accounts for 25%, then the maternal sex chromosomal gene of this group of seedling is ♀ XY, then A, B
Group all retains；It is respectively put into 1m2, depth 0.6m 30 mesh small-sized net cages cultivate, net cage be arranged in the preferable fish pond of water quality；Cultivation is extremely
After sexal maturity, fish female in A group is eliminated, remaining gives over to the male parent of experiment, and a tail tail is surveyed with Pelteobagrus vachelli mother fish ♀ XX respectively
It hands over, if filial generation male and female is fifty-fifty, male parent sex chromosome gene is ♂ XY；If filial generation is entirely milter, male parent gene is ♂ YY；It is female
Property filial generation cultivate the female parent tested to sexal maturity as test cross, respectively with Pelteobagrus vachelli milter ♂ XY carry out test cross, if sub
For half male and half female, then testing maternal sex chromosome gene is ♀ XX, must be eliminated；If filial generation female-male proportion is 1:3, mother is tested
This sex chromosome gene is ♀ XY, must also be eliminated；If all males of filial generation, testing maternal sex chromosome gene is
♀YY；The ♂ YY that test cross is selected mates breeding after cultivating with ♀ YY to sexal maturity, and filial generation is entirely Pelteobagrus vachelli supermale fish,
Spray is open the 6th day, takes out part seedling and carries out feminized treatment, and supporting to sexal maturity respectively without feminized treatment,
Mate group expanding, and group builds up high-quality Pelteobagrus vachelli supermale fish Germplasm Bank.By high-quality Pelteobagrus vachelli supermale fish ♂ YY and step (1)
The full raun ♀ XX mating breeding of the pure lines Pelteobagrus fulvidraco of acquisition can be obtained pure lines all-male hybridization Pelteobagrus fulvidraco ♂ XY.
As an improvement of the above technical solution, the spermatozoa preservative fluid the preparation method comprises the following steps: will analyze pure potassium chloride 0.4g,
Sodium chloride 7.8g, glucose 1.6g and 1,600,000 unit of penicillin are substantially soluble in 1000ml distilled water, are subsequently placed in 4 DEG C of refrigerators
It saves backup.
Further, the activation of spermatozoa liquid the preparation method comprises the following steps: sodium chloride 3g is dissolved in 1000ml tap water, shake up standby
With.
Further, the cold shock method are as follows: after artificial insemination 2min, fertilized eggs are moved into 4 DEG C of ice water and make cold shock
18min is handled, oxygenation hatching emergence in the normal incubation water for be put into 28 DEG C is then taken out.
Further, in the step (1) physiology milter ♂ XX selection are as follows: use the 17 Alpha-Methyl testis containing male sex hormone
After fairy shrimp 1h living is impregnated in the salt water inflation that ketone concentration is 100 μ g/L, the net worm of leaching feeds the full raun ♀ XX spray of pure lines, feeds day
Breakfast, lunch and dinner continuously feed 45 days, then continue to raise fry to sexal maturity with feed hormone-free, can be obtained physiology milter
♂XX。
Further, it is described containing 17 Alpha-Methyl testosterone concentration of male sex hormone be 100 μ g/L salt water the preparation method comprises the following steps: taking
17 Alpha-Methyl stosterone 100mg are dissolved in 98 ° of alcohol of 100ml, mix well, and obtain turning property stoste, take above-mentioned turning property stoste
10ml is added in 90ml98 ° of alcohol, mixes well, be made into dilution；It takes 1ml dilution to be added in 1 liter of salt water, can be obtained
The medical fluid for being 100 μ g/L containing 17 Alpha-Methyl testosterone concentration of male sex hormone.
The beneficial effects of the present invention are:
The invention belongs to a kind of pure lines all-male hybridization Pelteobagrus fulvidraco and its selections of parent.Using fish cell engineering heredity
Breeding technique is that Pelteobagrus fulvidraco and Pelteobagrus vachelli carry out continuous two generations' work gynogenesis to wild original, germplasm is purified
Rejuvenation, selects the full raun of pure lines Pelteobagrus fulvidraco and the pure lines full raun of Pelteobagrus vachelli, and to be sheerly Pelteobagrus vachelli as raw material fish,
Develop high-quality Pelteobagrus vachelli supermale fish, high-quality Pelteobagrus vachelli supermale fish hybridize with the pure lines full raun of Pelteobagrus fulvidraco produce it is pure
It is all-male cross hybrid seedling, thoroughly solves the various difficult and complicated cases that germplasm degeneration mixes and causes from parent source, eliminate sieve
Bottom " mao mao fish " parent significantly improves disease-resistant anti-adversity ability, the life of hybridization Pelteobagrus fulvidraco far from the probability of inbreeding seed
Long speed and bait conversion ratio, it is ensured that culture efficiency and food safety are Pelteobagrus fulvidraco industry along green, health, efficient, increasing
The Road Development of receipts opens a technical strategies, while providing sufficient pure lines Wa Shi for national Pelteobagrus fulvidraco hatchery
The parents such as supermale Pelteobagrus fulvidraco and the complete female Pelteobagrus fulvidraco of pure lines, large-scale production pure lines all-male hybridize advanced fry of yellow catfish, meet Pelteobagrus fulvidraco
Demand of the industry to pure lines all-male hybrid seedlings.
Detailed description of the invention
Fig. 1 is the schematic diagram of the technology of the present invention route.
Specific embodiment
The present invention will be further described in detail below with reference to the accompanying drawings and specific embodiments.
Referring to Fig.1, one embodiment of the present of invention: a kind of selection being sheerly all-male hybridization Pelteobagrus fulvidraco and its parent,
Concrete operation method with steps are as follows:
One: key of the invention is that wild original is that Pelteobagrus fulvidraco and Pelteobagrus vachelli apply cell engineering genetic breeding skill
Art repeats to purificate and rejuvenate to respective parent, and establishes the pure lines full raun of Pelteobagrus fulvidraco and the pure lines full raun of Pelteobagrus vachelli respectively
Germplasm Bank.
Wild original is Pelteobagrus fulvidraco and wild original is Pelteobagrus vachelli, fishes from " 800 inner Dongting Lake " lake region and chooses on fishing boat.
Pelteobagrus fulvidraco male and female individual reaches 150g and 350g or more respectively；Pelteobagrus vachelli male and female individual reaches 300g and 1500g or more respectively.This
The purification and rejuvenation of two kinds of fish parents select the completely homozygous pure lines of gene loci by independent continuous two generations gynogenesis
Full raun obtains physiology milter by hormone sex reversal, and physiology milter combines with full raun, hybridizes group expanding, establishes respectively pure
It is full raun Germplasm Bank.
1. being sheerly the foundation of the full raun Germplasm Bank of Pelteobagrus fulvidraco
(1) first generation gynogenesis
1. material and medicament
A. oxytocic hormone: chorionic gonadotrophin (HCG), luteinizing hormone releasing hormone, dioxone (DOM)；
B. male sex hormone: 17 α-Methyltestosterone (MT)；
C. pure potassium chloride 0.4g, sodium chloride 7.8g, glucose 1.6g, 1,600,000 unit of penicillin will spermatozoa preservative fluid: be analyzed
It is substantially soluble in 1000ml distilled water, is placed in 4 DEG C of refrigerators and saves；
D. activation of spermatozoa liquid: sodium chloride 3g is dissolved in 1000ml tap water, is shaken up spare；
E.95-98% alcohol and water for injection；
F corollary equipment: conventional hasten parturition tool, 2537 angstroms of wavelength of ultraviolet radiator, microoscillator；
G. different smart parent population: Pearl River system male Silurus Soldatovi Meridionalis fish
2. operating procedure
A. the preparation of the Heterologous Sperm of inhereditary material inactivation
Southern catfish milter is accelerated the ripening by the per kilogram of body weight LH injection μ of RH-A25-10 g, and smart 2g is manually taken after 10 hours, with guarantor
Liquid storage dilutes 3 times, stirs evenly and tiles on vessel, and thickness is no more than 0.5mm, is placed on microoscillator, starts oscillator, uses wave
Long 2537 angstroms of ultraviolet radiator makees radiation source, irradiates 25min；
B. the collection and insemination of fish-egg
Wild original is Pelteobagrus fulvidraco raun by per kilogram of body weight LH injection RH-A2 12 μ g, DOM 3mg, HCG 600IU, raw
Manage salt water 5ml；Parent population temporarily supports 28 DEG C of water temperature, and 12-15 hours extruding fish bellies collect fish-egg；Extract the sperm of appropriate genetic inactivation
It is added in fish-egg, mixes, add activation of spermatozoa liquid, stir evenly, insemination work is completed；
C. the induction of gynogenetic diploid
When artificial insemination, the Heterologous Sperm Y chromosome of genetic inactivation is not involved in the development of fertilized eggs, and only X chromosome is sent out
It educates and Haploid production development, and monoploid seedling can not survive, therefore, we use cold shock method, block subtrahend point
It splits second polar body discharge and induces gynogenetic diploid.Specific practice: fertilized eggs are moved into 4 DEG C of ice water by 2min after artificial insemination
In make cold shock treatment 18min, then take out in the normal incubation water for be put into 28 DEG C oxygenation hatching emergence.
Since the sexual control mechanism of Pelteobagrus fulvidraco is that male gamete is special-shaped, female gamete homotype, so ♀ XX gynogenesis
Filial generation is complete female filial generation.
(2) second generation gynogenesis
Since the high-quality wild original that first generation gynogenesis parent is the big water body in lake region is Pelteobagrus fulvidraco, purified through gynogenesis
Afterwards, gene loci is substantially homozygous, belongs to high-quality complete female filial generation.But in order to enable germplasm purer more excellent, we are by first generation thelykaryon
Development filial generation is cultivated to sexal maturity, carries out second generation gynogenesis according to the above method, selects gene loci homozygosis completely
It is sheerly Pelteobagrus fulvidraco female filial generation entirely.
(3) it is sheerly the foundation of the full raun Germplasm Bank of Pelteobagrus fulvidraco
The full raun ♀ XX of pure lines Pelteobagrus fulvidraco that second generation gynogenesis obtains, opening are taken out wherein 500 tails and are carried out after 6 days
Artificial turning property, obtains the physiology milter ♂ XX with normal reproduction ability.Full raun ♀ XX and physiology milter ♂ XX will be sheerly to cultivate
To sexal maturity, hybridize group expanding, establish the pure lines full raun Germplasm Bank of Pelteobagrus fulvidraco, mass production is sheerly the full raun of Pelteobagrus fulvidraco for big rule
Mould breeding pure lines all-male hybridization Pelteobagrus fulvidraco is used as female parent.
Specific practice is as follows:
A. the preparation of turning property stoste:
17 α-Methyltestosterone (MT), 100 μ g is taken, is dissolved in 98 ° of alcohol of 100ml, mixes well, male sex hormone stoste
1ml 1mg containing male sex hormone (i.e. 1000 μ g)；
B. the preparation of turning property dilution:
Above-mentioned stoste 10ml is taken, is added in 98 ° of alcohol of 90ml, mixes well, is made into dilution, 1ml dilution is containing male
100 μ g of hormone；
C. pharmaceutical chemistry is brewed:
Fairy shrimp 1h living is impregnated with containing the salt water inflation that 17 α of male sex hormone-Methyltestosterone (MT) concentration is 100 μ g/L
Afterwards, the net worm of leaching feeds fry, day feeds breakfast, lunch and dinner, continuously feeds 45 days.Then continue to raise fry with feed hormone-free
To sexal maturity, it is spare all to stamp electronic tag conservation.
D. it is sheerly full raun group expanding
Pure lines physiology milter ♂ XX mates with physiology raun ♀ XX, and mass propagation is sheerly full raun filial generation out, the special pool cultivate to
It is spare after sexal maturity.
2. being sheerly the foundation of the full raun Germplasm Bank of Pelteobagrus vachelli
Breeding be sheerly the full raun ♀ XX of Pelteobagrus vachelli, and establish pure lines the full raun Germplasm Bank of Wa Shi, technology path and
Operating process is identical as the technology and methods of the above-mentioned full raun of pure lines Pelteobagrus fulvidraco.
Two, the research and development of high-quality Pelteobagrus vachelli supermale fish and its foundation of Germplasm Bank
To be sheerly the full raun ♀ XX of Pelteobagrus vachelli and high-quality wild original is Pelteobagrus vachelli milter ♂ XY as raw material fish, answer
With genetic method and test cross, turning property technology, high-quality Pelteobagrus vachelli supermale fish is developed, and establish high-quality Pelteobagrus vachelli
Supermale fish Germplasm Bank is purified, the high-quality Pelteobagrus vachelli supermale fish ♂ YY of mass propagation by group expanding and germplasm, for large-scale production
It is sheerly all-male hybridization Pelteobagrus fulvidraco and lays in enough male parents.
The technology path that high-quality Pelteobagrus vachelli supermale fish and its Germplasm Bank are established, substantially by the applicant in 2016
Applied November 16, " it is complete that breeding is sheerly Pelteobagrus fulvidraco to the patent of invention of announcement on March 15th, 2017 (publication No. CN106489799A)
Raun and supermale fish large-scale production all-male fish method " disclosed in technology path breeding.The specific operation method is as follows:
1. the production of hybrid seeds of high-quality Pelteobagrus vachelli physiology raun
It chooses the well-developed pure lines full raun ♀ XX of Pelteobagrus vachelli and the especially excellent wild original of germplasm is Wa Shi Huang
Forehead fish milter ♂ XY mating breeding hatches high-quality Pelteobagrus vachelli filial generation ♀ XX and ♂ XY.Fry is open the 6th day, takes out 1000
Tail Pelteobagrus vachelli spray is independently cultivated, the good year after being impregnated with the 17-α ethinyl estradiols (EE2) that concentration is 150 μ g/L
Worm live bait carries out feeding, eats within one day 3, even feeds 50 days, fed later with conventional feed to sexal maturity.The Wa Shi Huang forehead of this approving and forwarding property
In fish, physiology raun and common raun respectively account for 50%.
2. test cross selects high-quality Pelteobagrus vachelli supermale fish ♂ YY and supermale physiology raun ♀ YY
It is directly cultivated in mature high-quality Pelteobagrus vachelli from above-mentioned, chooses 2-3 tails individual maximum, external physical characteristic is optimal
The most mature milter ♂ XY of good, gonad development, dissection are taken out spermary, are ground, and add and save the dilution sperm that liquid is made into 1:3, and sealing is put
It is saved backup into 4 DEG C of refrigerators.
From being cultivated in mature high-quality Pelteobagrus vachelli raun after above-mentioned turning property, 20 tails individual maximum, resemblance are chosen
The most mature pseudo- raun of fitst water, gonad development, squeezes out fish-egg respectively after hastening parturition, independently carry out people with preprepared sperm
Work insemination, fish-egg is uniformly layered in mesh sheet and is independently hatched.
After fry hatching membrane, the fry that each tail mother fish produces all is made a record, independent to cultivate, and cannot be mixed.Opening
After 6th day, the spray that this 20 tail parent population produces respectively is selected and remain into 200 tails, and is divided into two groups, every group of 100 tails, A group conventional feed is straight
Nursing is connect, B group pharmaceutical chemistry continues to raise after feminizing.
After raising 3 months, the gender of A group can be determined.If half male and half female, the maternal sex chromosomal gene of the group is
♀ XX eliminates two groups of A, B；If in A group filial generation, male accounts for 75%, and female accounts for 25%, then the maternal sex chromosomal gene of this group of seedling
For ♀ XY, retain two groups of A, B.
The 60 mesh small-sized net cages cultivation that filial generation is respectively put into 1m2, depth 0.6m will be retained, net cage is arranged in the preferable fish pond of water quality
In.
Cultivation is eliminated fish female in the filial generation of not turning property, remaining male parent tested as test cross to after sexal maturity, respectively with
Pelteobagrus vachelli mother's fish ♀ XX test cross, if filial generation male and female is fifty-fifty, male parent sex chromosome gene is ♂ XY；If filial generation is entirely hero
Fish, then male parent gene is ♂ YY.The filial generation feminized cultivates the female parent tested to sexal maturity as test cross, respectively with Wa Shi Huang forehead
Fish milter ♂ XY carries out test cross, if filial generation half male and half female, testing maternal sex chromosome gene is ♀ XX；If filial generation sex ration
Example is 1:3, then testing maternal sex chromosome gene is ♀ XY；If all males of filial generation, maternal sex chromosome is tested
Gene is ♀ YY.
The ♂ YY that test cross is selected mates breeding with ♀ YY, and filial generation is entirely Pelteobagrus vachelli supermale fish.When spray cultivate to
After 6th day, takes out part seedling and carry out feminized treatment, support to sexal maturity, mate group expanding with ♂ YY, and group builds up high-quality Wa Shi
Pelteobagrus fulvidraco supermale fish Germplasm Bank, the pure lines Pelteobagrus fulvidraco raun ♀ YY that high-quality Pelteobagrus vachelli supermale fish ♂ YY and step (1) are obtained
Mating breeding can be obtained pure lines all-male hybridization Pelteobagrus fulvidraco ♂ XY.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, as long as with essentially identical
Means realize that the technical solution of the object of the invention belongs within protection scope of the present invention.
Claims (6)
1. the selection of a kind of pure lines all-male hybridization Pelteobagrus fulvidraco and its parent, which comprises the following steps:
(1) the pure lines full raun Germplasm Bank of Pelteobagrus fulvidraco is established, is Pelteobagrus fulvidraco raun by continuous two generations gynogenesis by wild original, enables
Its gene loci is completely combined, and is generated and is sheerly complete female Pelteobagrus fulvidraco, and establishes the pure lines full raun Germplasm Bank of Pelteobagrus fulvidraco, wherein first
For gynogenesis the following steps are included:
A, the preparation of the Heterologous Sperm of inhereditary material inactivation, the southern catfish milter for being derived from Pearl River system is injected by per kilogram of body weight
LHRH-A2 5-10 μ g accelerates the ripening, and smart 2g is manually taken after 10 hours, dilutes 5 times with spermatozoa preservative fluid, stirs evenly and be laid in vierics
On ware, thickness is no more than 0.5mm, is placed on microoscillator, starts microoscillator, is made with 2537 angstroms of wavelength of ultraviolet radiator
Radiation source irradiates 25min, to obtain the Heterologous Sperm of genetic inactivation；
B, the collection and insemination of fish-egg will be Pelteobagrus fulvidraco raun from " 800 inner Dongting Lake " lake region wild original chosen on fishing boat of fishing
By per kilogram of body weight LH injection RH-A2 12 μ g, DOM 3mg, HCG 600IU, physiological saline 2ml；It is Jin Jiang River water that parent population, which temporarily supports water,
Library mid-water, water temperature are controlled to 28 DEG C, and fish belly is squeezed after 12-15 hours, collect fish-egg；Extract the heterologous of above-mentioned genetic inactivation
Sperm is added in fish-egg, mixes, adds activation of spermatozoa liquid, stir evenly, and insemination work is completed；
C, the induction of gynogenetic diploid, when above-mentioned artificial insemination, Heterologous Sperm is not involved in the development of fertilized eggs, only raun fish-egg
It develops and Haploid production, and monoploid seedling can not survive, for this purpose, blocking meiosis second by cold shock method
Polar body is discharged and induces gynogenetic diploid, and since the sexual control mechanism of Pelteobagrus fulvidraco is that male gamete is special-shaped, female gamete is same
Type, so ♀ XX Gynogenetic Offspring is complete female filial generation；
Second generation gynogenesis: first generation Gynogenetic Offspring is cultivated to sexal maturity, is carried out according to the above method female for the second time
Caryogenesis selects the completely homozygous pure lines Pelteobagrus fulvidraco female filial generation entirely of gene loci；
Be sheerly the foundation of the full raun Germplasm Bank of Pelteobagrus fulvidraco: the full raun ♀ XX of pure lines Pelteobagrus fulvidraco that second of gynogenesis obtains is opened
Mouth takes out wherein 500 tails and carries out artificial turning property, obtain the physiology milter ♂ XX with normal reproduction ability after 6 days；Pure lines are complete female
Fish ♀ XX and physiology milter ♂ XX are cultivated to sexal maturity, hybridize group expanding, establish the pure lines full raun Germplasm Bank of Pelteobagrus fulvidraco, great Liang Sheng
It is maternal for large-scale breeding pure lines all-male hybridization Pelteobagrus fulvidraco work to produce the pure lines full raun of Pelteobagrus fulvidraco；
(2) the pure lines full raun Germplasm Bank of Pelteobagrus vachelli is established, breeding is sheerly the full raun ♀ XX of Pelteobagrus vachelli, and establishes pure lines
The full raun Germplasm Bank of Wa Shi, operating process is identical as the above-mentioned full raun of pure lines Pelteobagrus fulvidraco, is only parent population by the wild former yellow forehead of system
It is Pelteobagrus vachelli raun that fish raun, which is changed to wild original,；
(3) research and development of high-quality Pelteobagrus vachelli supermale fish and its foundation of Germplasm Bank, comprising the following steps:
A, the well-developed full raun ♀ XX of pure lines Pelteobagrus vachelli and kind are chosen in the production of hybrid seeds of high-quality Pelteobagrus vachelli physiology raun
The especially outstanding wild original of matter is Pelteobagrus vachelli milter ♂ XY mating breeding, hatches high-quality Pelteobagrus vachelli filial generation ♀ XX and ♂
XY, fry are open the 6th day, take out 1000 tail Pelteobagrus vachelli sprays and independently cultivate, the 17-α acetylene for being 150 μ g/L with concentration
Fairy shrimp living bait after base estradiol impregnates 1 hour carries out feeding, eats within one day 3, even feeds 50 days, fed later with conventional feed
To sexal maturity, in the Pelteobagrus vachelli of this approving and forwarding property, physiology raun and common raun respectively account for 50%；
B, test cross selects high-quality Pelteobagrus vachelli supermale fish ♂ YY and supermale physiology raun ♀ YY, directly cultivates maturation from above-mentioned
In high-quality Pelteobagrus vachelli, 2-3 tails individual maximum, the milter ♂ XY that external physical characteristic is fitst water, gonad development is most mature, solution are chosen
Spermary is taken out, is ground, spermatozoa preservative fluid is added to be made into the dilution sperm of 1:3, sealing, which is put into 4 DEG C of refrigerators, to be saved backup；In addition
From being cultivated in mature high-quality Pelteobagrus vachelli raun after above-mentioned turning property, selection 20 tails individual is maximum, resemblance is fitst water, property
Gland develops most mature pseudo- raun, squeezes out fish-egg after hastening parturition respectively, independently carries out artificial insemination with above-mentioned ready sperm, will
Fish-egg, which is uniformly layered in mesh sheet, independently hatches；After fry hatching membrane, the fry that each tail mother fish produces all is made a record, independent
It cultivates, cannot mix；It is open after the 6th day, the spray that this 20 tail parent population produces arbitrarily is taken out into 200 tails, is divided into two groups of A, B, every group
100 tails, A group are directly fed with conventional feed, and B group pharmaceutical chemistry continues to raise after feminizing；After raising 3 months, statistics is each respectively
The male and female quantity of group, if half male and half female, the maternal sex chromosomal gene of the group is ♀ XX, eliminates A group and B group；If organizing a filial generation
In, male accounts for 75%, and female accounts for 25%, then the maternal sex chromosomal gene of this group of seedling is ♀ XY, retains two groups of A, B；It will retain
Filial generation is respectively put into 1m2, depth 0.6m 30 mesh small-sized net cages cultivate, net cage be arranged in the preferable fish pond of water quality；Cultivation to property at
After ripe, fish female in the filial generation of not turning property is eliminated, remaining is surveyed with Pelteobagrus vachelli mother fish ♀ XX respectively as the male parent tested
It hands over, if filial generation male and female is fifty-fifty, male parent sex chromosome gene is ♂ XY；If filial generation is entirely milter, male parent gene is ♂ YY；It is female
Property filial generation cultivate to sexal maturity as experiment female parent, respectively with Pelteobagrus vachelli milter ♂ XY carry out test cross, if filial generation is female
Male fifty-fifty, then testing maternal sex chromosome gene is ♀ XX；If filial generation female-male proportion is 1:3, maternal property dyeing is tested
Body gene is ♀ XY；If all males of filial generation, testing maternal sex chromosome gene is ♀ YY；♂ YY that test cross is selected with
♀ YY mating breeding, filial generation is entirely Pelteobagrus vachelli supermale fish, after cultivating to the 6th day, take out part seedling carry out it is female
Propertyization processing, is supported to sexal maturity, mate group expanding with ♂ YY, and group builds up high-quality Pelteobagrus vachelli supermale fish Germplasm Bank, by high-quality watt
Family name Pelteobagrus fulvidraco supermale fish ♂ YY mates breeding with the pure lines Pelteobagrus fulvidraco raun ♀ XX that step (1) obtains, and it is miscellaneous to can be obtained pure lines all-male
Hand over Pelteobagrus fulvidraco ♂ XY.
2. the selection of a kind of pure lines all-male hybridization Pelteobagrus fulvidraco according to claim 1 and its parent, it is characterised in that:
The spermatozoa preservative fluid the preparation method comprises the following steps: pure potassium chloride 0.4g, sodium chloride 7.8g, glucose 1.6g and penicillin will be analyzed
1600000 units are substantially soluble in 1000ml distilled water, are subsequently placed in 4 DEG C of refrigerators and are saved backup.
3. the selection of a kind of pure lines all-male hybridization Pelteobagrus fulvidraco according to claim 1 and its parent, it is characterised in that:
The activation of spermatozoa liquid the preparation method comprises the following steps: sodium chloride 2.5-3.5g is dissolved in 800-1200ml tap water, shake up spare.
4. the selection of a kind of pure lines all-male hybridization Pelteobagrus fulvidraco according to claim 1 and its parent, it is characterised in that:
The cold shock method are as follows: after artificial insemination 2min, fertilized eggs are moved into 4 DEG C of ice water and make cold shock treatment 18min, are then taken
It is put into oxygenation hatching emergence in 28 DEG C of normal incubation water out.
5. the selection of a kind of pure lines all-male hybridization Pelteobagrus fulvidraco according to any one of claims 1 to 4 and its parent, special
Sign is: the selection of physiology milter ♂ XX in the step (1) are as follows: is with the 17 Alpha-Methyl testosterone concentration containing male sex hormone
After fairy shrimp 1h living is impregnated in the salt water inflation of 100 μ g/L, the net worm of leaching feeds the full raun ♀ XX spray of pure lines, feeds breakfast, lunch and dinner day, even
It is continuous to feed 45 days, then continue to raise fry to sexal maturity with feed hormone-free, can be obtained physiology milter ♂ XX.
6. the selection of a kind of pure lines all-male hybridization Pelteobagrus fulvidraco according to claim 5 and its parent, it is characterised in that:
It is described containing 17 Alpha-Methyl testosterone concentration of male sex hormone be 100 μ g/L salt water the preparation method comprises the following steps: taking 17 Alpha-Methyl stosterones
500 μ g are dissolved in 98 ° of alcohol of 500ml, mix well, and obtain turning property stoste, take above-mentioned turning property stoste 10ml, and 90ml is added
It in 98 ° of alcohol, mixes well, is made into dilution, above-mentioned dilution is added in 900ml salt water, can be obtained containing male sex hormone
17 Alpha-Methyl testosterone concentrations are the salt water of 100 μ g/L.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811410043.1A CN109392775A (en) | 2018-11-23 | 2018-11-23 | A kind of selection being sheerly all-male hybridization Pelteobagrus fulvidraco and its parent |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811410043.1A CN109392775A (en) | 2018-11-23 | 2018-11-23 | A kind of selection being sheerly all-male hybridization Pelteobagrus fulvidraco and its parent |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109392775A true CN109392775A (en) | 2019-03-01 |
Family
ID=65455372
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811410043.1A Withdrawn CN109392775A (en) | 2018-11-23 | 2018-11-23 | A kind of selection being sheerly all-male hybridization Pelteobagrus fulvidraco and its parent |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109392775A (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111165401A (en) * | 2020-02-27 | 2020-05-19 | 广西壮族自治区水产引育种中心 | Rapid and efficient breeding method for tilapia |
| CN111328740A (en) * | 2020-03-17 | 2020-06-26 | 中国水产科学研究院长江水产研究所 | Breeding method for rapidly growing silver carps and application thereof |
| CN112790125A (en) * | 2021-01-22 | 2021-05-14 | 中国科学院水生生物研究所 | Creation method of rabbit hundred-pleated skirt Taishi goldfish |
| CN113661948A (en) * | 2021-09-06 | 2021-11-19 | 华中农业大学 | Method for obtaining full-female homozygous pelteobagrus fulvidraco clone line through self fertilization |
| CN116569885A (en) * | 2023-05-10 | 2023-08-11 | 华中农业大学 | Method for cultivating hybrid pelteobagrus fulvidraco yellow you No. 2 with fast growth, low protein, low fish meal feed utilization rate and high yield |
-
2018
- 2018-11-23 CN CN201811410043.1A patent/CN109392775A/en not_active Withdrawn
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111165401A (en) * | 2020-02-27 | 2020-05-19 | 广西壮族自治区水产引育种中心 | Rapid and efficient breeding method for tilapia |
| CN111165401B (en) * | 2020-02-27 | 2023-04-25 | 广西壮族自治区水产引育种中心 | Quick and efficient breeding method for tilapia mossambica |
| CN111328740A (en) * | 2020-03-17 | 2020-06-26 | 中国水产科学研究院长江水产研究所 | Breeding method for rapidly growing silver carps and application thereof |
| CN112790125A (en) * | 2021-01-22 | 2021-05-14 | 中国科学院水生生物研究所 | Creation method of rabbit hundred-pleated skirt Taishi goldfish |
| CN112790125B (en) * | 2021-01-22 | 2022-09-06 | 中国科学院水生生物研究所 | Method for creating rabbit-shaped goldfish with hundred-pleated skirt and Taishi |
| CN113661948A (en) * | 2021-09-06 | 2021-11-19 | 华中农业大学 | Method for obtaining full-female homozygous pelteobagrus fulvidraco clone line through self fertilization |
| CN116569885A (en) * | 2023-05-10 | 2023-08-11 | 华中农业大学 | Method for cultivating hybrid pelteobagrus fulvidraco yellow you No. 2 with fast growth, low protein, low fish meal feed utilization rate and high yield |
| CN116569885B (en) * | 2023-05-10 | 2024-01-23 | 华中农业大学 | Method for cultivating hybrid pelteobagrus fulvidraco yellow you No. 2 with fast growth and high feed utilization rate |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109392775A (en) | A kind of selection being sheerly all-male hybridization Pelteobagrus fulvidraco and its parent | |
| CN102047851B (en) | Construction and stock breeding method for cultured grass carp families | |
| CN101940184B (en) | Breeding and culturing method of improved crucian carp | |
| CN103960184B (en) | A kind of loach industrial artificial breeding improves the seedling-cultivating method of survival rate | |
| CN102349462A (en) | Breeding method for improved variety of fishes | |
| CN104642212A (en) | Artificial breeding method for onychostoma simus | |
| CN108377936B (en) | Large-scale production method of all-female mandarin fish | |
| CN103798173B (en) | For the progeny testing method of yellow cartfish and snakehead supermale fish and full milter exploitation | |
| CN101416613A (en) | The method of feminizing of WZ ♀-ZZ ♂ type Oreochromis aureus ZZ zygote | |
| CN107094671A (en) | A kind of all-male hybridizes the breeding method of Pelteobagrus fulvidraco | |
| CN105409856A (en) | Snakeheaded fish breeding method | |
| CN106879509A (en) | A kind of bar house oil fish artificial fecundation method | |
| CN106489799B (en) | The method that breeding is sheerly the full raun of Pelteobagrus fulvidraco and supermale fish large-scale production all-male fish | |
| CN1327763C (en) | Method of establishing bred prawn pedigree and selectively breeding improved prawn variety | |
| CN105265362A (en) | Cross breeding method for improving growth traits of slender mandarinfish | |
| CN100387120C (en) | A fish androgenesis method | |
| CN102812913B (en) | Cultivation method for hybridized all-female golden trout fries | |
| CN112262794A (en) | Method for sustainable large-scale production of all-female mandarin fish | |
| CN107155959A (en) | A kind of compound breeding method for improveing yellow catfish growing character | |
| CN107155971A (en) | Yi Zhong Gan fishes and black carp quasi aromatic hybrid rice breeding method | |
| CN101449664A (en) | Seed production method of WZ(male)-ZZ(female) all-male blue tilapia | |
| CN106614116A (en) | Method for inducing gynogenesis of Chinese sturgeons through homologous sperms | |
| CN109673545A (en) | A kind of red goldfish breeding cultural method of gooseneck boom | |
| CN108541632A (en) | A kind of sturgeon mating system | |
| CN104488759B (en) | A kind of method of molecule assist-breeding grass carp excellent strain and Breeding Effect checking |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20190301 |