CN101699998A - Selection and culture method of excellent breed large-tooth flounder featuring fast growth and high survival rate - Google Patents
Selection and culture method of excellent breed large-tooth flounder featuring fast growth and high survival rate Download PDFInfo
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Abstract
The invention relates to a selection and culture method of excellent breed large-tooth flounder featuring fast growth and high survival rate. The technical method comprises: culture and reproduction regulation and control of large-tooth flounder breeding group, establishment of family by sperm freezing technology, culture and standardization of the family, determination and analysis of number and properties, estimation of genetic parameters, screening of excellent family, and establishment of excellent breed by inducing thelykaryon of large-tooth flounder to develop by using heterologous frozen sperms. The invention can select and culture new breed of cultured large-tooth flounder featuring fast growth and high survival rate, screen out four excellent large-tooth flounder families featuring fast growth and high survival rate (0703, 0719, 0750 and 0751), and establish seven excellent breeds of large-tooth flounder developed from thelykaryon (0920, 0921, 0927, 0938, 0939, 0946 and 0947). The technical method can be applied to new variety culture of large-tooth flounder cultivation, solve the problems of culturing large-tooth flounder fish in sea water such as slow growth and low survival rate, and enhances economic benefit of large-tooth flounder cultivation.
Description
Technical field:
Present technique belongs to the marine biotechnology field, is a kind of breeding method of selecting to cultivate fast growth, the high lefteye flounder new lines of survival rate.
Background technology:
Lefteye flounder (Paralichthys olivaceus) is to be distributed in the coastal main economic fish of China, be naturally fish for, the main object of batch production, pond and cage culture, in fishery production, occupy bigger proportion.But along with the increasing to native species resource catching intensity, the decline of lefteye flounder germ plasm resource manifests.Propagating artificially under the environment in addition, China does not obtain enough attention to the lefteye flounder good variety selection for a long time, inbreeding breeding, intensive culture etc. cause the degeneration of lefteye flounder breed variety germplasm, growth rate to reduce, the culture-cycle is long, breed efficient is low, have had a strong impact on the development of popularization of lefteye flounder seed cultivating large area and industrialization.Therefore utilize the disease-resistance population of different geographical populations of lefteye flounder and artificial screening to carry out the lefteye flounder breed of variety, cultivation fast growth, the lefteye flounder that resistance against disadvantage is strong are cultured new varieties, and be significant for the recovery of lefteye flounder germplasm, raising cultured product quality and economic benefit.Aspect the research of lefteye flounder breed new varieties, screenings such as Kanako Fuji cultivate one can disease-resistant lymphocyst disease cultured population (Faculty of Marine Science, Tokyo University ofMarine Science and Technology, Kanako Fuji et al, Aquaculture, 2006).This seminar utilizes microsatellite marker and AFLP technology that the genetic structure of lefteye flounder cultured population and wild population is estimated (Inst of Huanghai Sea Marine Products, Chinese Academy of Aquatic Product Science in recent years, Liu et al, Aquaculture, 2005), find that the genetic diversity of cultured population obviously reduces.The disease-resistant fish of lefteye flounder and not the MHC II B gene pleiomorphism of disease-resistant fish and the correlation of anti-vibrio anguillarum disease have been analyzed, (China Aquatic Science Research Institute's Huanghai Sea produces research institute to Preliminary screening to disease-resistant relevant MHC II B genotype, Zhang etal, Marine Biotechnology, 2006), (China Aquatic Science Research Institute's Huanghai Sea produces research institute to utilize 3 breeding populations to set up the lefteye flounder family simultaneously, Chen Songlin, field Yongsheng etc., aquatic product journal, 2008), from then on launched domestic lefteye flounder breeding work, but fast in growth, the selection cultivation aspect of the high lefteye flounder new lines of survival rate at home and abroad also belongs to first.
Summary of the invention:
The objective of the invention is at propagating under the environment the slow and low problem of survival rate of lefteye flounder seed growth rate artificially, cultivate fast growth, lefteye flounder new lines that survival rate is high, thus raising lefteye flounder culturing economic benefit.
The present invention realizes by following technical method:
Content of the present invention mainly comprises: 1, the lefteye flounder breeding population is cultivated and genital regulating, 2, the auxiliary family of sperm freezing technology is set up, 3, family is cultivated and standardization, 4, the measurement of quantitative character and analysis, 5, genetic parameter is estimated, 6, superior families screening, 7, utilize the allos frozen sperm to induce the lefteye flounder gynogenesis to make up excellent strain.
1, the lefteye flounder breeding population is cultivated and genital regulating:
Following 3 lefteye flounder breeding populations are utilized the electronic mark mark respectively, set up the breeding archives,, reach 2-3 age propagating concentrated raising cultivation under the condition artificially.The year of breeding population cultivates temperature and is controlled at 13-25 ℃, adopts normal sunshine, and water temperature mating season of the annual 2-5 month is controlled at 14-17 ℃, light application time 15h, and intensity of illumination 1500-2000lux, the Continuous irradiation 1-2 month, the current exchange capacity is 6-7m
3/ h, parent population utilizes commercially available artificial matched doped materials to feed at anestrous season, utilizes mating season the wild fishes that adds vitamin E to feed.Reach sexual maturity through above cultivation, but the output mature egg.4 breeding populations comprise: 1. breeding for disease resistance colony (RS): utilize the mode of artificial infection Vibrio anguillarum to screen individuality 100 tails of not falling ill for a long time behind the bacteria infection from cultured population and form.2. Japanese breeding population (JS): introduce the lefteye flounder fry from Japan and form through artificial culture, screening 30 tails.3. Huanghai Sea breeding population (YS): form from individual 50 tails of the natural wild lefteye flounder of fishing for of Yantai, the Shandong Province coastal collection in area.
2, the auxiliary family of sperm freezing technology is set up:
Utilize manual compression belly method, gather ripe lefteye flounder parent population seminal fluid, utilize MPRS+20%DMOS solution with 1: 1 dilution proportion of volume ratio, inject the 2.0ml cryovial, put into the freezing preservation of liquid nitrogen, each constituent concentration of MPRS solution is: NaCl 60.35mM, NaH
2PO
41.8mM; NaHCO
33mM, KCl 5.23mM, CaCl
2.2H
2O 1.13mM, MgCl
2.6H
2O 1.13mM, D-Glucose 55.55mM.DMSO is commercially available dimethyl sulfoxide (DMSO).
Utilize the sperm of above 3 breeding populations of the freezing preservation of above method and the ripe milter of screening superior families, and collect and the freezing lefteye flounder milter sperm of having preserved from other breed company, set up lefteye flounder breeding population sperm freezing storehouse in advance, utilize frozen sperm indirect labor fertilization.Between the parent, adopt 1 mating patterns male how female or 1 female many heros to set up the lefteye flounder family between 3 breeding populations or in the colony.
3, family is cultivated and standardization:
Utilize the controllable cycle water of water quality, discharge, temperature and illumination workshop to carry out family hatching, seed rearing, the cultivation water body of each family is 3m
3Water vat, discharge can be at 1-5m
3Regulate and control between/the h, water temperature can be 14-25 ℃ of regulation and control, and illumination can keep 10-15h.Utilize chlorella, wheel animalcule and halogen worm to carry out the cultivation of early stage fry, utilize Articial bait making to carry out the cultivation of advanced fry.Grow to 15,50,90,150 fry respectively, the breed quantity of each family carried out standardization about 210d, standardization fry quantity is respectively 10000,1000,500 and 200 tails, and 210d concentrates the standardization fry and raises at 1-2 30m
3In the pond, utilize red, orange, yellow 3 kinds of fluorescent pigments simultaneously, before the fry belly is selected the back of the body, in the back of the body, behind, 5 positions behind the abdomen and in the abdomen, utilize injection system that selection family individuality is carried out mark.
4, quantitative character is measured and is analyzed:
The family growth traits is measured: the Measuring Time of family quantitative character is: grow to several time periods in 90,210,510,720 (2 ages), 90-210 days is that the lefteye flounder family is set up first high temperature season of back, also is first fast growing period of lefteye flounder fry; 210-510 days is first survive the winter phase and second fast growing period of fry, and fry had been spent second winter in 720 days, and most of individuality has reached marketable fish size, and the fast individuality of partly growing has reached sexual maturity.Measure body long (holding to tail fin end), body weight and the breed survival quantity of fry in succession from kiss in each period.
Character analysis: long, the weight data of the body that utilizes lefteye flounder family different growing stage to measure, adopt one-way analysis of variance and Student-Newman-Keuls (SNK) method that different family growth traits are carried out multiple comparative analysis, calculate growth mean value and standard deviation simultaneously.Utilize absolute weightening finish formula AGR
W(g.d
-1)=(W
2-W
1)/(t
2-t
1) calculate each family absolute rate of body weight gain of growth regular period.Utilize (fry survive quantity/standardization after sum) * 100% computist to be tied to form motility rate.Analyze the growth differences between family, select the fastest and high family of survival rate of growth.
5, the estimation of genetic parameter:
Utilize lefteye flounder family body length, the weight data of different growing stage mensuration, adopt additivity and dominant variance component, components of covariance, genetic correlation and the heritability of linear mixed model (Linear mixed model), " additivity-dominance " genetic analysis model and constraint maximum-likelihood method (REML) estimated body length and weight character.Additivity and dominant variance reach utmost point significance level (P＜0.01), genetic correlation reaches extremely significantly positive correlation level (P＜0.01), heritability reaches extremely significantly (P＜0.01) and greater than 0.2, explanation selection effect long to body and body weight is obvious, detect with this and to select the effect that inherent cause and environmental factor take place in the breeding process, select breeding for lefteye flounder relevant parameter is provided.
6, superior families screening:
By mensuration and comparative analysis to family growth traits after the standardization, from the family of setting up, filter out body length and body weight and have significant difference (P＜0.05), the absolute rate of body weight gain of 90-524d is more than 0.25, survival rate is in the family more than 80%, and, survival rate high family fast as growth continues to cultivate.Utilize mark to separate with other family from concentrate to culture the pond family that filters out, specially the pond is cultivated to 2-3 and is reached sexual maturity age, and breeding method is as described in the content 1, and on each individuality the permanent electronics mark of injection, the recorded electronic sign is set up the breeding archives.
7, utilize the allos frozen sperm to induce the lefteye flounder gynogenesis to make up excellent strain:
Gather ripe perch sperm in advance, utilize MPRS+20%DMOS solution with 1: 1 dilution proportion perch sperm of volume ratio, adopt the freezing preservation in liquid nitrogen of 2.0ml cryovial, set up perch sperm freezing storehouse, each constituent concentration of MPRS solution is: NaCl 60.35mM, NaH
2PO
41.8mM; NaHCO
33mM, KCl 5.23mM, CaCl
2.2H
2O 1.13mM, MgCl
2.6H
2O 1.13mM, D-Glucose55.55mM.DMSO is commercially available dimethyl sulfoxide (DMSO).
The superior families of screening is cultivated to sexual maturity, reached the requirement of laying eggs, utilize manual compression belly method to gather unfertilized egg by artificial genital regulating.Utilizing 37 ℃ of water-baths to separate the frozen sea-bass sperm thereupon, utilize the MPRS dilution with the dilution proportion of sperm with 1: 10, is to add 1ml dilution seminal fluid in the culture dish of 10cm to pave at diameter, at 40000 μ J/cm
2The intensity ultraviolet ray is irradiation down, make its genetic material inactivation, add and carry out the dry method insemination in the lefteye flounder ovum, fertilized egg is 100: 0.4 with the seminal fluid ratio, immerse cold shock processing 45min in 2～4 ℃ of seawater behind the insemination 3min, make its chromosome doubling, then ovum is put into 17 ℃ of seawater hatchings, so far finish good lefteye flounder family gynogenesis, set up excellent strain.
The technical characterstic of technical solution of the present invention: 1, utilized lefteye flounder breeding population Japan's lefteye flounder and the Huanghai Sea lefteye flounder that has typical geographical features in the lefteye flounder distribution of west bank, the Pacific Ocean, and artificial infection's method made up disease-resistant lefteye flounder colony, enriched the genetic background of lefteye flounder good variety selection.2, the freezing preservation technology of fish sperm is fused in the foundation of lefteye flounder family, for the hybridization assembly provides abundant halibut spermatozoon source and more group mode, improved family and set up efficient.3, utilize the freezing perch sperm of allos to induce the individual gynogenesis of lefteye flounder superior families, improved the speed of isozygotying of superior families gene, 1 generation gynogenesis suitable 4 generation full sibs brother and sister selfings, simultaneously the allos sperm induces the homozygosity that has guaranteed gynogenesis offspring gene.4, the measurement of foundation by a large amount of familys and screening, quantitative character and multiple ratio, the estimation and the gynogenesis method screening of genetic parameter and set up fast growth lefteye flounder new lines 0719,0750 and 0751.5, modern biotechnologies such as sperm freezing technology, gynogenesis technology, genetics of quantitative characters parametric estimate and traditional selection breeding are combined, improved the efficient of breeding of new variety, set up the new way of a breeding.
Embodiment:
The principles of science of technical solution of the present invention institute foundation: modern biotechnology combined with traditional selection breeding technique carry out rearing new variety.Selecting breeding (selective breding) is according to breeding objective, in the natural variation type that occurs in existing kind or breeding material, through relatively identifying, by the multiple choices method, select excellent roguing, select good variation individuality, cultivate the method for new varieties.Family selective breeding is one of method of selecting breeding, by foundation, breeding, the comparison of family, eliminates bad family, selects standard compliant family, and the hereditary homozygosity by generation raising kind can select the new varieties with stable merit.The technology of the present invention is preserved sperm freezing modern biotechnology such as technology, gynogenesis technology, genetic parameter estimation and traditional selection breeding technique combines, and improves breeding level, has significantly accelerated fish rearing new variety process.
Be described in detail technology contents of the present invention below by embodiment:
Content of the present invention mainly comprises: 1, the lefteye flounder breeding population is cultivated and genital regulating, 2, the auxiliary family of sperm freezing technology is set up, 3, family is cultivated and standardization, 4, the measurement of quantitative character and analysis, 5, genetic parameter is estimated, 6, superior families screening, 7, utilize the allos frozen sperm to induce the lefteye flounder gynogenesis to make up excellent strain.
1, the lefteye flounder breeding population is cultivated and genital regulating:
Utilize the electronic particle injection to carry out mark 3 lefteye flounder breeding populations, and write down its electronic mark, simultaneously to its body is long, body weight is measured and record, sets up the breeding population archives.3 breeding populations comprise: 1. breeding for disease resistance colony (RS): utilize the mode of artificial infection Vibrio anguillarum to screen individuality 100 tails of not falling ill for a long time behind the bacteria infection from cultured population and form.2. Japanese breeding population (JS): introduce the lefteye flounder fry from Japan and form through artificial culture, screening 30 tails.3. Huanghai Sea breeding population (YS): form from individual 50 tails of the natural wild lefteye flounder of fishing for of Yantai, the Shandong Province coastal collection in area.
Reach 2-3 age through artificial culture, the year of breeding population cultivates temperature and is controlled at 13-25 ℃, adopts normal sunshine, water temperature mating season of the annual 2-5 month is controlled at 14-17 ℃, light application time 15h, intensity of illumination 1500-2000lux, the Continuous irradiation 1-2 month, the current exchange capacity is 6-7m
3/ h, parent population the season of growth with the artificial mixed feed of throwing something and feeding, feed breeding period and add the wild fishes of vitamin E, through above growth and genital regulating, reach sexual maturity and artificial propagation requirement, manual compression fish body belly can flow out ripe ovum and sperm.
2, the auxiliary family of sperm freezing technology is set up:
Halibut spermatozoon freezing and storing method: utilize manual compression belly method to gather ripe halibut spermatozoon, the microscopic examination sperm viability, vigor utilizes MPRS+20%DMOS solution with 1: 1 dilution proportion of volume ratio 70% above person, adopts the freezing preservation in liquid nitrogen of 2.0ml cryovial.Surplus the halibut spermatozoon 500 of freezing each breeding population of preservation part (about 900ml), set up the halibut spermatozoon freezer altogether.When artificial propagation, the halibut spermatozoon of freezing preservation is thawed in 37 ℃ of water-baths, microscopy freezes the back sperm viability, remain on more than 70% as sperm viability, can with the lefteye flounder ovum dry method fertilization of gathering, the smart ratio of the time of fertilization ovum is: 100ml: 1ml.
Lefteye flounder method for establishing stemma: utilize above 3 breeding populations, select sexual maturity individuality in the colony, artificial ovum and the sperm (or in the freezer sperm) gathered adopts following mating pattern to set up half sibs or family full-sibs: RS ♂ * JS ♀, RS ♂ * RS ♀, RS ♂ * YS ♀, JS ♂ * RS ♀, JS ♂ * JS ♀, JS ♂ * YS ♀, YS ♂ * RS ♀, YS ♂ * JS ♀ in 3 breeding populations or between colony.(table 1).
Table 1 2007 lefteye flounder family pedigrees
Annotate: RS: disease-resistant lefteye flounder, JS: Japanese lefteye flounder, YS: Huanghai Sea lefteye flounder
3, the cultivation of family and standardization:
Family is cultivated: lefteye flounder family fry is cultivated at 3m
3In the water vat, adopt miniflow water in early days, every day exchange capacity 1-2m
3, keep water quality pure and fresh, every day, each utilized the algae of precipitating at the bottom of the siphonage service sink, superfluous feed for 1 time sooner or later.3m
3Put newly hatched larvae quantity in the water vat in a suitable place to breed about 10000 tails, volt ground fish seedling (the long 1cm of body) 2000 tails, 5-7cm fry 1000 tails, 10-15cm fry 500 tails.The concentration of chlorella remains on 8～100,000/ml in the water body, and the wheel animalcule feeding volume is advisable with 5-10 in the water/ml, and halogen worm feeding volume is advisable with 1-2/ml.Throwing something and feeding of artificial feed should in time be regulated the feed granules size with fry specification size, granular mixed feed began to throw something and feed in 12-15 days, seed pellet particle diameter 250-400 μ m before 25 days, 0.1-0.15g the young postlarva pellet particle diameter 400-600 μ m of body weight is about 0.5g body weight pellet particle diameter 800 μ m.Along with the growth of fish body, the manufactured feed particle diameter increases gradually.It is the 5-15% of fish body weight that manufactured feed day is thrown the amount of raising, and throws something and feeds in time, should lack and throw diligent the throwing.Prevent that phenomenons such as feed surplus, variations in temperature, water pollution, flow instability from taking place.
The family standardization: the family difference that the standardized main purpose of family causes breeding environment is reduced to minimum.Family is cultured and to about 15 days fry density tentatively to be adjusted, and every nautilus seedling quantity is remained on about 10000 tails, cultures and all familys is carried out the standardization first time in 50 days, each family 2000 tail fishes of selecting and remain at random.(5-7cm) 1000 tails of selecting and remain about 90 days, (10-15cm) 500 tails of selecting and remain continue to culture about 150 days.Culture about 210 days selection 200 tail individualities at random from each family, utilize red, yellow, 3 kind fluorescent pigments of orange, before the fry belly is selected the back of the body, in the back of the body, behind, mark is carried out in 5 positions behind the abdomen and in the abdomen, concentrates raising at 1-2 30m behind the mark
3In the pond, long to the body of selecting the family individuality simultaneously, body weight is measured one by one, finishes the standardization of family quantity and breeding environment this moment.
4, the measurement of quantitative character and analysis:
Setting up the lefteye flounder family in 2007 is growing into about average age in days to 90,524d, body length, body weight, survival rate to each family are measured, utilize one-way analysis of variance and Student-Newman-Keuls (SNK) method that different family growth traits are carried out multiple ratio, analyze the growth differences between family.Utilize absolute rate of body weight gain (AGR
W, g.d
-1)=(W
2-W
1)/(t
2-t
1), the absolute rate of body weight gain of each family growth 90d to 524d is calculated average age in days (t
2-t
1) press 286d and calculate, survival rate=(524d family survival quantity/200 tails) * 100%, filter out the family that growth is fast, survival rate is high according to absolute rate of body weight gain and breed survival rate, 0719,0750,0703, No. 0751 family growth the fastest survival rate the highest (table 2).
Table 2 lefteye flounder family in 2007 90d and 524d growth traits are relatively
Annotate: * has significant difference (P＜0.05) aspect growth
5, the estimation of genetic parameter:
Utilize 2007 and set up 63 familys, collected the long and weight data of body of all family 5328 tail fries. by the genetic correlation and the heritability of linear mixed model (Linear mixed model), " additivity-dominance " model, constraint maximum-likelihood method (REML) difference estimated body length and body weight.The result shows that body additivity, dominance, phenotype and genotype correlated components long and body weight two proterties all reach utmost point significance level (P＜0.01), the additive variance ratio of body length and body weight is all greater than the dominant variance ratio, and the hereditary effect of visible two proterties depends primarily on the additive effect of gene.Body length and body weight reach extremely significantly positive correlation (P＜0.01).The long total hereditary effect rate of body is 31.20%, total hereditary effect rate of body weight is 33.58%, the body ratio of variance at random long and body weight reaches 68.80% and 66.41% respectively, illustrate that the influence long to the lefteye flounder body and body weight of factor such as breeding environment is bigger, illustrate simultaneously at lefteye flounder and select in the breeding elimination environmental influence very important.Body narrow heribatility long and body weight is respectively 0.20 and 0.26 (P＜0.01), and broad heritability is respectively 0.31 and 0.34 (P＜0.01), and lefteye flounder belongs to medium heritability on above two proterties, can obtain bigger genetic progress (table 3) by selecting breeding.
Table 3: utilize " additivity-dominance " model to lefteye flounder family body length and body weight component of variance, component of variance ratio and heritability estimated result
Annotate: 0** represents significance P＜0.01
6, the screening of superior families:
Reaching genetic parameter through mensuration, multiple ratio to family cultivation in 2007, growth, survival rate proterties estimates, filtered out had that growth is fast in 2007 in the family, totally 4 of the lefteye flounder familys of the high proterties of survival rate: be respectively 0719,0703,0750 and 0751 (table 2, table 4), absolute rate of body weight gain is 0.27-0.372, and survival rate is 82.5-100%.Through artificial culture, genital regulating to the screening family, cultivate and regulate and control method as described in the specific implementation method 1, spring in 2009, part family individuality reached sexual maturity, induced by the allos frozen sperm and had set up gynogenesis strain (table 5).
Table 4 family The selection result in 2007
Annotate: before the 1-back of the body, the 2-behind, behind the 3-abdomen, the Y-yellow, O-is orange, the R-redness
7, utilize the allos frozen sperm to induce the lefteye flounder gynogenesis to make up excellent strain:
Artificial culture, the genital regulating of, survival rate high family 0719,0750,0751 fast by growth that 2007 are filtered out reach sexual maturity in April, 2009, utilize extruding belly method can collect unfertilized egg.Simultaneously, set up its sperm bank, utilized 37 ℃ of water-baths sperm that thaws, utilized the MPRS dilution with the dilution proportion of sperm, at 40000 μ J/cm with 1: 10 at the time of fertilization in freezing preservation of in November, 2008 perch sperm 200ml
2The intensity ultraviolet ray is irradiation down, makes its genetic material inactivation.Gather the individual ovum of above ripe family, the perch sperm of inactivation is added in the ovum, carry out the dry method insemination, the smart ratio of the time of fertilization ovum is 100: 0.4, put into 2-4 ℃ of seawater cold shock behind the insemination 3min and handle 45min, put into 17 ℃ of seawater hatchings and fry rearing then.Setting up above family gynogenesis in succession is 7: be respectively 0921,0927,0920,0938,0939,0946 and 0947 (table 5).The growth of gynogenesis system is measured, and gynogenesis is the long 10.54-12.28cm of being of body at present, and body weight is 13.19-20.62g.
Table 5 lefteye flounder gynogenesis excellent strain pedigree and growing state
Annotate: the disease-resistant lefteye flounder of RS-, JS-Japan lefteye flounder, the perch frozen sperm of S.P-deactivation
Claims (1)
1. the selection of a large-tooth flounder featuring fast growth and high survival rate excellent strain, it is characterized in that its method, comprising: mensuration and analysis, the genetic parameter of the cultivation of the cultivation of lefteye flounder breeding population and genital regulating, the auxiliary family foundation of sperm freezing technology, family and standardization, quantitative character estimated, superior families screens, utilize the allos frozen sperm to induce the lefteye flounder gynogenesis to make up excellent strain; The disease-resistant lefteye flounder, Japanese lefteye flounder, the Huanghai Sea lefteye flounder that make up are in advance utilized the electronic particle injection of labelled, and concentrated cultivation reaches 2-3 age, and a year cultivation temperature is controlled at 13-25 ℃, the artifical compound feed of throwing something and feeding; The annual 2-5 water temperature mating season month is controlled at 14-17 ℃, light application time 15h, and intensity of illumination 1500-2000lux, the Continuous irradiation 1-2 month, the current exchange capacity is 6-7m
3/ h, the wild fishes that adds vitamin E of feeding is cultivated, and when reaching sexual maturity, manually gathers ripe lefteye flounder ovum and sperm, and the microscopy vigor is kept in the liquid nitrogen at the sperm freezing more than 70%, sets up the halibut spermatozoon freezer; During artificial propagation frozen sperm is thawed in 37 ℃ of water-baths, the microscopy vigor is in 70% above person and the fertilization of ovum dry method, and the smart ratio of its ovum is 100ml: 1ml; With the mating patterns of 1 male how female or 1 female many heros, set up full sibs or family half sibs between 3 breeding populations or in the colony; Family before the 210d is cultivated at 3m
3In the water vat, discharge is regulated and control at 1-5m
3Between/the h, water temperature 14-25 ℃; Grow to 15,50,90,150 fry respectively, the breed quantity of each family carried out standardization about 210d, standardization fry quantity is respectively 10000,1000,500 and 200 tails, that 210d utilizes respectively is red, yellow, 3 kinds of fluorescent pigments of orange, before the fry belly is selected the back of the body, in the back of the body, behind, 5 positions select individuality to carry out mark to family behind the abdomen and in the abdomen, and concentrate breed at 1-2 30m
3In the pond; Family grows to 90,210,510, several time periods of 720d are long to its body, body weight and survival rate are measured, utilize variance analysis, multiple ratio, definitely weightening finish and survival rate comparative analysis family growing state; Adopt linear mixed model to estimate measuring genetic parameters, body narrow heribatility long and body weight is respectively 0.20 and 0.26 (P＜0.01), and broad heritability is respectively 0.31 and 0.34 (P＜0.01); By above method filter out that growth is fast, 4 of high lefteye flounder familys of survival rate: be respectively family 0703,0719,0750, No. 0751.To screen family and cultivate maturation, but the output mature egg; The perch sperm of freezing preservation is thawed in 37 ℃ of water-baths, and with the dilution proportion perch seminal fluid of MPRS with 1: 10, the back is at 40000 μ J/cm
2Ultraviolet ray irradiation down, make its genetic material inactivation, add and carry out the dry method insemination in the lefteye flounder ovum, fertilized egg and seminal fluid volume ratio are 100ml: 0.4ml, immerse cold shock processing 45min in 2～4 ℃ of seawater behind the insemination 3min, induce lefteye flounder ovum gynogenesis, then ovum is put into 17 ℃ of seawater hatchings, set up grow fast, the high lefteye flounder strain of survival rate, upright 7 excellent strains of building together are respectively strain 0920,0921,0927,0938,0939,0946, No. 0947.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104920255A (en) * | 2015-06-01 | 2015-09-23 | 中国水产科学研究院黄海水产研究所 | F3-generation rapid-growth flounder pure line construction method |
| CN105638538A (en) * | 2016-03-30 | 2016-06-08 | 全国水产技术推广总站 | Bastard halibut ecological marking method for release counting |
| CN106035144A (en) * | 2016-05-30 | 2016-10-26 | 中国水产科学研究院黄海水产研究所 | Disease resistance evaluation method of cultured shrimps and crabs |
| CN106577382A (en) * | 2016-11-29 | 2017-04-26 | 浙江省海洋水产研究所 | Pseudosciaenapolyactis family establishment and superior family selection method |
| CN112931312A (en) * | 2021-02-26 | 2021-06-11 | 海南晨海水产有限公司 | Artificial breeding method of seriolala quinqueradiata |
| CN113317274A (en) * | 2021-06-17 | 2021-08-31 | 河北科技大学 | Method and equipment for accurately screening growth genetic characteristics of breeding pigs under colony cultivation |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107211927A (en) * | 2017-06-06 | 2017-09-29 | 舟山市普陀兴海养殖优质种苗选育研究所 | A kind of selective breeding method for disease-resistant superior strains of paralichthys olivaceus |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101305701B (en) * | 2008-07-08 | 2011-01-26 | 中国水产科学研究院黄海水产研究所 | Fish family establishing and disease-resistant high yield fine breeding method |
-
2009
- 2009-11-03 CN CN2009102300122A patent/CN101699998B/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104920255A (en) * | 2015-06-01 | 2015-09-23 | 中国水产科学研究院黄海水产研究所 | F3-generation rapid-growth flounder pure line construction method |
| CN105638538A (en) * | 2016-03-30 | 2016-06-08 | 全国水产技术推广总站 | Bastard halibut ecological marking method for release counting |
| CN106035144A (en) * | 2016-05-30 | 2016-10-26 | 中国水产科学研究院黄海水产研究所 | Disease resistance evaluation method of cultured shrimps and crabs |
| CN106035144B (en) * | 2016-05-30 | 2019-04-02 | 中国水产科学研究院黄海水产研究所 | A kind of disease resistance trait appraisal procedure cultivating shrimp crab |
| CN106577382A (en) * | 2016-11-29 | 2017-04-26 | 浙江省海洋水产研究所 | Pseudosciaenapolyactis family establishment and superior family selection method |
| CN106577382B (en) * | 2016-11-29 | 2019-08-20 | 浙江省海洋水产研究所 | A kind of building of little yellow croaker family and breeding superior families method |
| CN112931312A (en) * | 2021-02-26 | 2021-06-11 | 海南晨海水产有限公司 | Artificial breeding method of seriolala quinqueradiata |
| CN113317274A (en) * | 2021-06-17 | 2021-08-31 | 河北科技大学 | Method and equipment for accurately screening growth genetic characteristics of breeding pigs under colony cultivation |
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| CN101699998B (en) | 2011-10-12 |
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