CN102090360B - High-efficient induction method of tetraploid cynoglossus semilaevis fish fries - Google Patents
High-efficient induction method of tetraploid cynoglossus semilaevis fish fries Download PDFInfo
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Abstract
The invention relates to a high-efficient induction method of tetraploid cynoglossus semilaevis fish fries, and the method comprises the steps of induction of the tetraploid cynoglossus semilaevis fish fries and identification of the tetraploid cynoglossus semilaevis fish fries, and is characterized by performing pressure shock treatment for 3.5-6 minutes under hydrostatic pressure of 38-44MPa within 19-28 minutes after fertilization; and transferring fertilized eggs into seawater at the temperature of 19-23 DEG C for culture and incubation after completing the shock treatment. According to the invention, a technical method which can be used for effectively inducing the fertilized eggs of cynoglossus semilaevis to produce tetraploid fish fries in a doubled manner is established, and a technical parameter for inhibiting the first cleavage of the eggs of the cynoglossus semilaevis is firstly determined. The method is simple to operate, strong in practicality, easy to implement, safe and reliable; and the induction efficiency of the tetraploid fish fries is high. By adopting the method, a new technical approach is provided for the sex control and polyploid breeding of the cynoglossus semilaevis.
Description
Technical field
The invention belongs to aquatic products genetic breeding technical field, be specifically related to a kind of tetraploid Cynoglossus semilaevis fry highly effective revulsion induction method, promptly adopt biological technique method to induce the Cynoglossus semilaevis tetraploid.
Background technology
Cynoglossus semilaevis (Cynoglossus semiliaevis) belongs to Pleuronectiformes (Pleuronectiformes), Cynoglossidae (Cynoglossidae), tongue sole genus (Cynoglossus); Popular name sole rice, dragon profit, tongue fish etc. are the distinctive famous and precious economic seawater fishs of China.Because its delicious flavour, fine and tender taste, nutritious are welcome by consumers in general deeply, its market value is high.Cynoglossus semilaevis cultivation is generally carried out in China's Coastal Areas, and its aquaculture annual value of production reaches hundred million yuans of 15-20.But; Because Cynoglossus semilaevis male and female difference in size great disparity; The male growth rate has slowly reduced cultured output, and big specification raun gonad development is excessive, and belly is obviously protruding; Influence marketable fish quality and attractive in appearance, thereby had a strong impact on the enthusiasm of fisherman's breed and the development of cynoglossus semilaevis cultivation industry.Triploid fish is formed unevenly because of chromosome, and sexual gland can not be reached maturity, and has avoided the protruding problem of raun belly, thus faster than the liploid fish growth, commodity value is higher, more get consumer reception.Therefore, adopt the modern biotechnology means, research Cynoglossus semilaevis multiploid induction technology; Develop tetraploid fish, with the sterile triploid fish of a large amount of productions of liploid fish mating, can reach the inhibition gonad development then; Gonad is grown the energy that consumes be used for growth; Degradation problems under the growth retardation that overcomes gonad development period and the flesh of fish quality, this improves cultured output and economic benefit for shortening the cynoglossus semilaevis cultivation cycle; Promote the development of cynoglossus semilaevis cultivation industry, have important practical significance and great application value.
In view of significance and the using value of fish multiploid induction technology in genetic breeding, fish production, water environment protection and the protection of aquatile genetic diversity, also pay much attention to the sterile Study on Technology of aquiculture animal such as fish in the world.And make the sterile main path of fish is to produce triploid.Because the triploid fish have the control excessive multiplication, suppress gonad development, promote advantages such as growth and raising flesh quality, thereby receive general attention.Most research work of fish polyploid are carried out on freshwater fish.Fish triploids such as grass carp, channel catfish, Tilapia mossambica, rainbow trout, coho, chinook, Atlantic salmon, silver carp and carp have successively been obtained so far; In addition, in Britain and Japan, the triploid rainbow trout has also got into the commercialization breed.Except that rainbow trout, Japan has also carried out the research and development of induced triploid sweetfish, finds that growth rate and the cold tolerance of triploid sweetfish all obviously is superior to dliploid sweetfish, receives very much consumer and the welcome of culturing house, has very big commercial value and potentiality to be exploited.But, above-mentioned research all is to adopt the cold shock method to make chromosome doubling through suppressing the second polar body discharge, directly induced triploid.Because triploid induction rate is lower, the triploid fry limited amount of production does not reach the purpose of commercially producing, and therefore, directly the method for induced triploid is difficult to reach the requirement of extensive industrialization.And through inducing tetraploid,, can produce sterile triploid fry in a large number, this point important that on Cynoglossus semilaevis, seems then with the dliploid mating.But, up to now, the mass of relevant Cynoglossus semilaevis tetraploid fry is induced, and produces the sterile fry of triploid etc. in a large number through tetraploid and dliploid mating, does not appear in the newspapers as yet both at home and abroad.Particularly how long carrying out hydrostatic pressing at the Cynoglossus semilaevis ovum after fertilization handles, adopts great pressure, handles on the key technologies such as how long could obtaining the tetraploid fry and all do not obtain breaking through all the time.
Summary of the invention
The purpose of this invention is to provide a kind of tetraploid Cynoglossus semilaevis fry highly effective revulsion induction method, promptly, chromosome doubling is induced in a large number produce the tetraploid fry through suppressing the spilting of an egg for the first time of Cynoglossus semilaevis fertilized egg.The present invention set up the employing hydrostatic pressing and induced the tetraploid technical method of Cynoglossus semilaevis, broken through the technical barrier that exists in the Cynoglossus semilaevis tetraploid breeding for a long time, thereby filled up the blank of Cynoglossus semilaevis tetraploid induction.
Content of the present invention comprises two aspects: the inducing of (one) tetraploid Cynoglossus semilaevis fry; (2) evaluation of tetraploid Cynoglossus semilaevis fry.
(1) tetraploid Cynoglossus semilaevis fry induce step following:
1) Cynoglossus semilaevis essence, ovum collection and insemination:
Select sexually matured Cynoglossus semilaevis parent population, ovum is adopted in the dry beaker, the fish-egg of gathering is placed 19-23 ℃ of preservation; Gather fresh Cynoglossus semilaevis seminal fluid; The seminal fluid of gathering is added in the fish-egg, and the volume ratio of seminal fluid and fish-egg is 1: 100-1000, shake mixing gently after; The seawater that adds 2 times of temperature to the fish-egg volume and be 19-23 ℃ is accomplished fertilization process, after the insemination ovum is placed in the 19-23 ℃ of seawater and hatches;
2) chromosome doubling operation:
After insemination in the 19-28min, fertilized egg is placed the hydraulic cylinder of hydrostatic press, under the 36-44MPa hydrostatic pressing, to carry out the pressure shock and handle, the shock processing time is 3.5-6min, after shock disposes fertilized egg is moved into to cultivate in 19-23 ℃ the seawater and hatches;
Wherein the chromosome doubling zero-time is definite:
After insemination 19.0; 21.5,23.5,25.5 and 28.0 minutes different time; The model that places Japan to produce fertilized egg is the hydraulic cylinder of 5506 hydrostatic press; Under the 36-44MPa hydrostatic pressing, carry out pressure shock and handle, the shock processing time is 3.5-6min, after shock disposes fertilized egg is moved into to cultivate in 19-23 ℃ the seawater and hatches; Calculate fertilization rate mid-term at primitive gut, behind the fry hatching membrane, calculate incubation rate and the tetraploid rate of fry.According to incubation rate and the tetraploid ratio of each group fry, confirmed that effective zero-time that Cynoglossus semilaevis tetraploid fry is induced is insemination back 21.5-25.5min.
Hydrostatic pressing is induced confirming of Cynoglossus semilaevis tetraploid shock pressure and processing time:
After confirming hydrostatic pressing shock zero-time, set the hydrostatic pressing barometric gradient and be 36,38,40,42 and 44MPa, continue to handle 3.5-4.5 minute, after shock disposes ovum moved in the 23-24 ℃ of seawater and cultivate.Calculate fertilization rate mid-term at primitive gut; Behind the fry hatching membrane, calculate the incubation rate of tetraploid fry, the experimental result surface; Fertilized egg is 38; 40 can both obtain a certain proportion of tetraploid fry with 42MPa pressure, and induce the ratio of tetraploid fry the highest at hydrostatic pressing pressure 40 during with 42MPa, and suitable hydrostatic pressing pressure is 40-42MPa therefore to confirm the Cynoglossus semilaevis tetraploid induction.
Handle the duration according to last two experimental results screening hydrostatic pressing at last, gradient scope was located at 4-6 minute, carry out the hydrostatic pressing shock and handle.The counting offspring in the tetraploid production, experimental result shows, 21.5-25.5min after insemination, with Cynoglossus semilaevis fertilized egg under 40MPa pressure, suffer a shock the processing 4-5.5 minute, fry can both incubation of membrane.And when shock was handled 4.5-5.5 minute, the ratio of tetraploid fry was the highest.Therefore the suitable hydrostatic pressing processing time is 4.5-5.5 minute to confirm the Cynoglossus semilaevis tetraploid induction.
(2) evaluation of tetraploid Cynoglossus semilaevis fry
1) identify Cynoglossus semilaevis tetraploid fry with the ploidy analyzer:
Adopt the ploidy analyzer and the one-step method staining reagent of PARTEC company, measure tetraploid fry dna content, compare with the normal diploid fry.If the cell DNA content of certain bar fry is about two times of dliploid contrast fry cell DNA content, show that then this tail fish is a tetraploid.
2) chromosome analysis is identified Cynoglossus semilaevis tetraploid fry:
Gather the tetraploid induction fry, carry out chromosome analysis according to conventional method, counting chromosome quantity, the chromosome number of Cynoglossus semilaevis diploid fish fry is 42, if the chromosome quantity of certain bar fry is about 84, proves that then this fish is the tetraploid fry.
The present invention has set up and can effectively induce the Cynoglossus semilaevis fertilized ovum chromosome to double to produce the technical method of tetraploid fry; Confirmed to suppress the technical parameter of the Cynoglossus semilaevis ovum spilting of an egg for the first time first.Multiploid induction method is in the past compared, and the tetraploid fry abductive approach that the present invention sets up can be induced the tetraploid fry in a large number.Method of the present invention row simple to operate, practical, easy, safe and reliable; The tetraploid fry induce efficient higher.The tetraploid fry inductivity that this method induces is high, ploidy is neat, inductivity is stable, and therefore, this method is very effectively with reliable; For Cynoglossus semilaevis sex controlling and polyploid breeding have been opened up new technological approaches.
Description of drawings
Fig. 1: diploid fish fry cell DNA content.
Fig. 2: tetraploid fry cell DNA content.
Fig. 3: normal diploid fry chromosome division phase.
Fig. 4: tetraploid fry chromosome division phase of the present invention.
Embodiment
The present invention adopts the modern biotechnology means, has set up Cynoglossus semilaevis tetraploid induction technology, has developed the tetraploid fry.
Adopt the fertilization of homology sperm and Cynoglossus semilaevis ovum, certain hour after insemination carries out the hydrostatic pressing shock to fertilized egg and handles, and suppresses the spilting of an egg for the first time, makes chromosome doubling, can successfully induce the generation of Cynoglossus semilaevis tetraploid fry.Set up Cynoglossus semilaevis tetraploid fish height of seedling and imitate inductive technology; Produce for Cynoglossus semilaevis sex controlling, sterile triploid seed; Carry out sterile seed and culture, improve cultured output, product quality and the economic benefit of this Cynoglossus semilaevis, have important practical significance and the huge potentiality of applying.
Be elaborated in the face of technology contents of the present invention down:
Its technology contents of the present invention comprises two aspects: one, Cynoglossus semilaevis tetraploid fry induces; Two, the evaluation of Cynoglossus semilaevis tetraploid fry.
One, inducing of Cynoglossus semilaevis tetraploid fry:
1) Cynoglossus semilaevis essence, ovum collection and insemination
Select sexually matured Cynoglossus semilaevis parent population, or adopt the artificial induced spawning method to induce the raun ovulation, in 30-38 hour, manual compression raun belly is adopted ovum behind artificial induced spawning, and ovum is adopted in the dry beaker, and the fish-egg of gathering is placed 19-23 ℃ of preservation; Gather fresh Cynoglossus semilaevis seminal fluid through extruding sexual maturity milter belly; The seminal fluid of gathering is added in the unfertilized egg; The dry method insemination; Shake mixing gently, the volume ratio of seminal fluid and ovum is 500 μ L:50-100ml, and then to add 2 times of temperature to ovum amount volume be that 19-23 ℃ seawater 100 or 200ml accomplishes fertilization process.Fertilization is carried out in the 1000ml beaker.After fertilization is placed on ovum in the 19-23 ℃ of seawater hatches;
2) chromosome doubling operation
At first be confirming of chromosome doubling zero-time:
After insemination 19.5; 21.5,23.5,25.5 and 27.5 minutes different time; The model that places Japan to produce on the insemination ovum is the hydraulic cylinder of 5506 hydrostatic press; Under the 38-40MPa hydrostatic pressing, carry out pressure shock and handle, the shock processing time is 3.5-4.5min, suffers a shock ovum to be moved in the 19-23 ℃ of seawater after disposing and cultivates.Calculate fertilization rate mid-term at primitive gut, behind the fry hatching membrane, calculate the incubation rate of tetraploid fry.Carried out hydrostatic pressing handled at after fertilization 19.5-27.5 minute; Each group all has the tetraploid fry of 50-68% to produce; Wherein in the time of 21.5-25.5 minute, the fry hatching rate is the highest, and the ratio of acquisition tetraploid fry is also the highest; Reach 60-68%, confirm that therefore effective zero-time that Cynoglossus semilaevis tetraploid fry is induced is insemination back 21.5-25.5min (seeing table 1).
Table 1: hydrostatic pressing is induced confirm (ocean temperature is about 23 ℃, hydrostatic pressure 40MPa, shock processing time 4.5min) of Cynoglossus semilaevis tetraploid zero-time
| The insemination back time (min) | Incubation rate (is calculated fertilized egg, %) behind the 12AF | Tetraploid rate (%) |
| 19.5 | 4.4±1.0 | 51.3±4.7 |
| 21.5 | 16.7±1.2 | 68.3±4.4 |
| 23.5 | 10.8±1.7 | 61.7±6.0 |
| 25.5 | 9.4±0.9 | 60.7±5.2 |
| 27.5 | 3.6±0.5 | 52.0±4.6 |
| Control group | 56.1±7.2 | 0.0±0.0 |
Hydrostatic pressing is induced confirming of Cynoglossus semilaevis tetraploid shock pressure and processing time:
After confirming hydrostatic pressing shock zero-time, set the hydrostatic pressing barometric gradient and be 36,38,40,42 and 44MPa, continue to handle 3.5-4.5 minute, after shock disposes ovum moved in the 23-24 ℃ of seawater and cultivate.Calculate fertilization rate mid-term at primitive gut; Behind the fry hatching membrane, calculate the incubation rate of tetraploid fry, the experimental result surface; Fertilized egg is 38; 40 can both obtain a certain proportion of tetraploid fry with 42MPa pressure, and only when hydrostatic pressing pressure 40-42MPa tetraploid fry ratio the highest, therefore suitable hydrostatic pressing pressure is 40-42MPa to confirm the Cynoglossus semilaevis tetraploid induction.(seeing table 2)
Handle the duration according to last two experimental results screening hydrostatic pressing at last, gradient scope was located at 4-6 minute, carry out the hydrostatic pressing shock and handle.The counting offspring in the tetraploid production, experimental result shows, 21.5-25.5min after insemination, with Cynoglossus semilaevis fertilized egg under 40-42MPa pressure, suffer a shock the processing 4-5.5 minute, fry can both incubation of membrane.And when shock is handled 4.5-5.5 minute, induce the ratio of tetraploid fry the highest.(seeing table 3)
Table 2: hydrostatic pressing is induced confirm (about 23 ℃ of ocean temperatures, pressure duration is 4.5min) of Cynoglossus semilaevis tetraploid shock pressure
Table 3: hydrostatic pressing is induced the confirming of Cynoglossus semilaevis tetraploid duration (about 23 ℃ of ocean temperatures, after fertilization 21.5min, hydrostatic pressure is 40MPa)
| The hydrostatic pressure duration (min) | Incubation rate (is calculated fertilization behind the 12AF | Tetraploid rate (%) |
| Ovum, %) | ||
| 4 | 16.1±1.0 | 28.2±2.9 |
| 4.5 | 13.4±1.8 | 64.2±3.0 |
| 5 | 7.4±1.3 | 68.1±4.3 |
| 5.5 | 1.1±0.2 | 74.3±3.1 |
| 6 | 0.0±0.0 | 0.0±0.0 |
| Control group | 48.9±4.9 | 0.0±0.0 |
4. hydrostatic pressing induces Cynoglossus semilaevis tetraploid fry result to gather
The method of setting up above adopting, we have carried out the mass of Cynoglossus semilaevis tetraploid fry and have induced, and in 3 tetraploid inductions experiments, obtain fry 5.3 ten thousand tails, existing Cynoglossus semilaevis tetraploid induction result are gathered as follows
Table 4: the Cynoglossus semilaevis tetraploid induction is the result gather
Two, the evaluation of Cynoglossus semilaevis tetraploid fry:
Its technology contents comprises: identify Cynoglossus semilaevis tetraploid fry through the ploidy analyzer 1.; 2. identify the tetraploid fry through chromosome analysis.
1. identify Cynoglossus semilaevis tetraploid fry with the ploidy analyzer:
Each tetraploid induction experiment, 2-5 days tetraploid experiment fry 20-30 tail behind the sampling observation incubation of membrane.Every tail fry is put into the centrifuge tube of 1 1.5ml, clean fry 1 time, add 0.2ml PBS to each centrifuge tube subsequently, fry is processed single cell suspension with distilled water.In each pipe, add the one-step method staining reagent that 1-2ml buys from PARTEC company; Behind membrane filtration, filtrating is changed in the supporting 5ml test tube of instrument, flick test tube; Cell suspension is mixed, place PARTEC ploidy analyzer (PA) to measure in test tube then.Specimen, the embryo compares with normal diploid, analyzes tetraploid induction group fry again.
Through the cell DNA content analysis; Learn that the dna content overwhelming majority in the diploid fish fry cell concentrates on the 26-28 place, and the dna content in the most cells of tetraploid fry is at 52-56 (table 5), in the 30 tail artificial inductions' that detect tetraploid fry; The dna content that 19 tail fries are arranged is 52-56; Be 2 times (seeing Fig. 1, Fig. 2) of diploid fish fry, therefore prove that the fry that we induce is the tetraploid fry, the tetraploid induction rate is 63%.Adopting the ploidy analyzer mainly is feasibility and the validity that proof the inventive method is induced Cynoglossus semilaevis tetraploid fry, and when adopting the inventive method to carry out the Cynoglossus semilaevis tetraploid induction from now on, then unnecessary each detection of tetraploid fry of all carrying out.
Table 5: cells were tested by flow cytometry tetraploid Cynoglossus semilaevis cell DNA relative amount gathers
2. chromosome analysis is identified the tetraploid fry:
The chromosome number of Cynoglossus semilaevis diploid fish fry is the 2n=42 bar; And the chromosome number of tetraploid fry should be 4n=84 in theory, therefore can identify through chromosome analysis whether the fry that induces is the tetraploid fry.If the chromosome number of the fry that induces is 84, prove that then these fries are tetraploids.The present invention adopts the fry chromosome flaking method, and Cynoglossus semilaevis artificial induction's tetraploid fry is carried out sampling Detection.Key step comprises: 2-8 days fries are placed 0.02% colchicine of seawater configuration, at room temperature handled 2 hours.Put into fry hypotonic 30 minutes of the KCL of 0.075mol/L then.Ka Nuoshi liquid (methyl alcohol: glacial acetic acid=3: 1) fix 3 times each 20 minutes continuously with new preparation.Get single fry again and put into 50% glacial acetic acid, process cell suspension with sharp tweezers.Heat is dripped the sheet method and is dripped sheet.10% Giemsa stain dyeing 20-30 minute, microscopy.The common fry of Cynoglossus semilaevis is a dliploid, and its chromosome number is 42 (see figure 3)s; If the chromosome number of selective examination fry is 84, show that this fish is a tetraploid, if chromosome quantity is 42, then be diploid fish fry.Detect tetraploid ratio in the tetraploid induction experiment fry thus.Each tetraploid induction experiment sampling observation 10-20 tail fry carries out chromosome analysis; Individuality in the fry that discovery produces under the hydrostatic pressing shock condition of optimizing more than 60% contains 84 chromosome (see figure 4)s; Prove that these fries are tetraploids, its inductivity is about 60%.Adopting chromosome analysis mainly is that proof the inventive method is induced tetraploid feasibility of Cynoglossus semilaevis and validity, and when adopting the inventive method to carry out the Cynoglossus semilaevis tetraploid induction from now on, the then unnecessary chromosome analysis that at every turn all carries out the tetraploid fry.But after the fry sexual maturity to be induced, determine whether to be tetraploid through detecting reproductive cell.
Claims (5)
1. a tetraploid Cynoglossus semilaevis fry highly effective revulsion induction method comprises the steps:
1) Cynoglossus semilaevis essence, ovum collection and insemination:
Select sexually matured Cynoglossus semilaevis parent population, ovum is adopted in the dry beaker, the fish-egg of gathering is placed 19-23 ℃ of preservation; Gather fresh Cynoglossus semilaevis seminal fluid; The seminal fluid of gathering is added in the fish-egg, and the volume ratio of seminal fluid and fish-egg is 1: 100-1000, shake mixing gently after; Add 2 times to the fish-egg volume, temperature is that 19-23 ℃ seawater is accomplished fertilization process, after the insemination ovum is placed in the 19-23 ℃ of seawater and hatches;
2) chromosome doubling operation:
After insemination in the 19-28min; The hydraulic cylinder that the 19-23 ℃ of fertilized egg in the seawater places the hydrostatic press that is placed on step 1); Under the 40-42MPa hydrostatic pressing, carrying out the pressure shock handles; The shock processing time is 4.5-5.5min, after shock disposes fertilized egg is moved into and cultivates hatching in 19-23 ℃ the seawater;
3) evaluation of Cynoglossus semilaevis tetraploid fry:
The fry that hatches is carried out Methods of Ploidy Identification, filter out tetraploid Cynoglossus semilaevis fry.
2. abductive approach as claimed in claim 1 is characterized in that above-mentioned pressure shock processing 21.5-25.5min after insemination carries out.
3. abductive approach as claimed in claim 1 is characterized in that above-mentioned Methods of Ploidy Identification is to adopt ploidy analyzer analysis of cells dna content or adopt chromosome analysis method counting chromosome quantity.
4. abductive approach as claimed in claim 3 is characterized in that the above-mentioned cell DNA content that uses the tetraploid Cynoglossus semilaevis fry that the ploidy analyzer analyzes two times as dliploid Cynoglossus semilaevis fry cell DNA content.
5. abductive approach as claimed in claim 3 is characterized in that using the chromosome quantity of the definite tetraploid Cynoglossus semilaevis fry of chromosome analysis method is 84.
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| CN104322408A (en) * | 2014-10-17 | 2015-02-04 | 中山大学 | Siniperca chuatsi polyploid induction method |
| CN106900610B (en) * | 2017-02-14 | 2019-10-01 | 中国水产科学研究院黑龙江水产研究所 | Pneumatic type fish spilting of an egg generating means and cleavage stage seedling incubating method |
| CN107494358B (en) * | 2017-09-28 | 2019-11-01 | 中国科学院南海海洋研究所 | A kind of preparation method of Hong Kong oyster tetraploid children shellfish |
Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1263704A (en) * | 1999-02-15 | 2000-08-23 | 湖南师范大学 | Method for breeding tetraploid fish |
| CN1709046A (en) * | 2005-06-10 | 2005-12-21 | 湖南师范大学 | Red crucian, Japanese white crucian and megalobrame amblycephala subfamily distant hydridization method |
| CN1739346A (en) * | 2004-08-25 | 2006-03-01 | 中国科学院海洋研究所 | The method that the spilting of an egg for the first time of a kind of lefteye flounder fertilized egg suppresses |
| CN1861017A (en) * | 2005-10-28 | 2006-11-15 | 白宝海 | Heat-shock and static water-pressure type method for prodn. of tetraploid of trout |
| CN1969616A (en) * | 2005-11-25 | 2007-05-30 | 李思发 | Method for constructing allotetraploid of bluntsnout bream |
| CN101077063A (en) * | 2006-05-26 | 2007-11-28 | 中国科学院海洋研究所 | Method for breeding triploid monomer oyster in scale |
| CN101103707A (en) * | 2007-08-16 | 2008-01-16 | 大连水产学院 | Method for inducing and cultivating tetraploid breeding of sea urchin |
| CN101595849A (en) * | 2009-06-23 | 2009-12-09 | 中国水产科学研究院黄海水产研究所 | Flounder tetraplont fry batch induction method |
| CN101112186B (en) * | 2007-09-14 | 2011-02-09 | 湖南师范大学 | Method for cultivating high-back type crucian |
-
2010
- 2010-12-17 CN CN2010105943511A patent/CN102090360B/en not_active Expired - Fee Related
Patent Citations (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1263704A (en) * | 1999-02-15 | 2000-08-23 | 湖南师范大学 | Method for breeding tetraploid fish |
| CN1739346A (en) * | 2004-08-25 | 2006-03-01 | 中国科学院海洋研究所 | The method that the spilting of an egg for the first time of a kind of lefteye flounder fertilized egg suppresses |
| CN1709046A (en) * | 2005-06-10 | 2005-12-21 | 湖南师范大学 | Red crucian, Japanese white crucian and megalobrame amblycephala subfamily distant hydridization method |
| CN1861017A (en) * | 2005-10-28 | 2006-11-15 | 白宝海 | Heat-shock and static water-pressure type method for prodn. of tetraploid of trout |
| CN1969616A (en) * | 2005-11-25 | 2007-05-30 | 李思发 | Method for constructing allotetraploid of bluntsnout bream |
| CN101077063A (en) * | 2006-05-26 | 2007-11-28 | 中国科学院海洋研究所 | Method for breeding triploid monomer oyster in scale |
| CN101103707A (en) * | 2007-08-16 | 2008-01-16 | 大连水产学院 | Method for inducing and cultivating tetraploid breeding of sea urchin |
| CN101112186B (en) * | 2007-09-14 | 2011-02-09 | 湖南师范大学 | Method for cultivating high-back type crucian |
| CN101595849A (en) * | 2009-06-23 | 2009-12-09 | 中国水产科学研究院黄海水产研究所 | Flounder tetraplont fry batch induction method |
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