CN1476751A - Induction method of gynogenesis of multicalor abalone - Google Patents
Induction method of gynogenesis of multicalor abalone Download PDFInfo
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- CN1476751A CN1476751A CNA031329365A CN03132936A CN1476751A CN 1476751 A CN1476751 A CN 1476751A CN A031329365 A CNA031329365 A CN A031329365A CN 03132936 A CN03132936 A CN 03132936A CN 1476751 A CN1476751 A CN 1476751A
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- ovum
- haliotis diversicolor
- gynogenesis
- gynogenetic
- sperm
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Abstract
The method for inducing variegated abalone gynogenesis includes the following steps: firstly, using UV ray to irradiate sperm of variegated abalone to inactivate the genetic material of sperm of variegated abalone, then regulating the concentration of genetic inactivated sperm, making it mix with normal variegated abalone ovum, promoting gynogenesis of variegated abalone ovum, then using cytochalasin B or 6-dimethylaminopurine to inhibit second meiotic division of activated ovum to make genome be reduplicated to produce gynogenesis diploid, then transferring the treated variegated abalone gynogenesis diploid ovum into nursery container, making management according to conventional breeding method so as to obtain the invented variegated abalone gynogenesis diploid larva.
Description
(1) technical field
The present invention relates to the gynogenetic abductive approach of a kind of shellfish seedling gynogenetic abductive approach, especially Haliotis diversicolor.
(2) background technology
Mariculture shellfish seedling carries out genetic breeding by the Bao gynogenesis is induced in producing, and the offspring of its generation is only contained maternal instinct parent's gene, but the fast purifying germplasm is accelerated breeding speed.(Japanese aquatic products annual meeting will such as Arai, 1984,50:2019-2023) reported application ultraviolet irradiation inactivation haliotis discus hannai Ino sperm genetic substance, and with the gynogenesis of genetic inactivation activation of spermatozoa haliotis discus hannai Ino ovum, cultivate the haliotis discus hannai Ino gynogenetic haploid young, but, can't further grow because the thelykaryon monoploid young is highly lopsided at all.(Japanese aquatic products annual meeting will such as Fujino, 1990,56:1755-1763) reported the abductive approach of haliotis discus hannai Ino gynogenesis diploid:, induce gynogenesis ovum chromosome set to double with the method for cold shock more earlier with ultraviolet inactivation haliotis discus hannai Ino staining of sperm body.Use this method Fujino etc. and induced the haliotis discus hannai Ino gynogenesis diploid, but induce offspring's survival rate extremely low, 3.3%～9.0%, 6 monthly age survival rates only 0.005% are just only arranged in the veliger stage; Inductivity also only has 50%～60% simultaneously.
Haliotis diversicolor is the important abalone culture kind of China, though this kind belongs to Bao Ke together with haliotis discus hannai Ino, two kinds have than big difference: 1) 2 kinds have in heredity than big-difference, and the haliotis discus hannai Ino dliploid has 36 chromosomes, and Haliotis diversicolor only has 32; 2) 2 kind gamete forms have than big-difference, and the haliotis discus hannai Ino sperm belongs to transiens, and the Haliotis diversicolor sperm belongs to modified.At present, the gynogenesis of Haliotis diversicolor research still belongs to blank.
(3) summary of the invention
The object of the present invention is to provide the high gynogenetic abductive approach of Haliotis diversicolor of a kind of offspring's of inducing survival rate.
The gynogenetic abductive approach of the said Haliotis diversicolor of the present invention is as follows:
1) genetic inactivation of sperm: with ultraviolet radiation Haliotis diversicolor seminal fluid, ultraviolet irradiating dose is 60000～120000 μ W/cm
2, seminal fluid thickness 1～2mm, sperm concentration 10
6～10
8Individual/ml;
2) the gynogenetic startup of ovum: the Haliotis diversicolor sperm of Haliotis diversicolor mature egg and process ultraviolet irradiation is mixed, and the control temperature is at 23～30 ℃, and the sperm ultimate density is no less than 10
5Individual/ml, the gynogenesis of startup Haliotis diversicolor ovum;
3) the gynogenetic haploid chromosome set doubles;
4) the Haliotis diversicolor gynogenesis diploid ovum after the above-mentioned processing is changed in the container for plant growth,, produce Haliotis diversicolor gynogenesis diploid seedling by common Haliotis diversicolor seedling production method management.
Said gynogenetic haploid chromosome set doubles available cytochalasin B (Cytochalasin B in step 3), letter is CB) handle or with 6-dimethylaminopurine (6-dimethylaminopurine, letter is 6-DMAP) handle, suppress to activate the 2nd reduction division of ovum;
Said CB handles: ovum is transferred in the solution that contains 0.4～1.0mg/L cytochalasin B after starting 7～20 minutes, makes Haliotis diversicolor gynogenetic haploid chromosome set double to become gynogenesis diploid; Gynogenesis Haliotis diversicolor ovum after will doubling again to handle is transferred to and is embathed in 0.033%～0.05% dimethyl sulphoxide solution 2 times, each 10～15 minutes, removes residual cytochalasin B;
Said 6-DMAP handles: after ovum starts 7～20 minutes, transfer in the solution of 250～350 μ mol/L 6-dimethylaminopurines (6-dimethylaminopurine, letter is 6-DMAP) 10～20 minutes;
The present invention at first uses ultraviolet radiation Haliotis diversicolor seminal fluid, and the genetic material of inactivation Haliotis diversicolor sperm suppresses to activate the 2nd reduction division of ovum again with cytochalasin B or 6-dimethylaminopurine, chromosome set is doubled, and produces gynogenesis diploid.This method is induced offspring's survival rate height, and veliger stage survival rate reaches 99.5%, is used for the survival rate (3.3%～9.0%) of the abductive approach of haliotis discus hannai Ino with the stage young far above Fujino etc.; And the dliploid inductivity is up to 95%～100%, and the ovum starting rate is higher than 36%～49% ovum starting rate in the haliotis discus hannai Ino gynogenesis of reports such as Arai up to 96%～100%.
(4) embodiment
Embodiment 1
1. the genetic inactivation of sperm: get the Haliotis diversicolor seminal fluid through ultraviolet irradiation, ultraviolet irradiating dose is 90000 μ W/cm
2, seminal fluid thickness 1.5mm, sperm concentration 10
8Individual/ml.
2. the gynogenetic startup of ovum: the seawater water temperature is adjusted to 25 ± 1 ℃, mix the Haliotis diversicolor sperm of Haliotis diversicolor mature egg and process ultraviolet irradiation in this seawater, the sperm ultimate density is at least 10
5Individual/ml, the gynogenesis of startup Haliotis diversicolor ovum.Sediments microscope inspection, the ovum starting rate reaches 96%～100%.
3. the gynogenetic haploid chromosome set doubles: after ovum starts 20 minutes, transfer in the seawater that contains 0.7 ± 0.1mg/L cytochalasin B 20 minutes, make Haliotis diversicolor gynogenetic haploid chromosome set double to become gynogenesis diploid; Gynogenesis Haliotis diversicolor ovum after will doubling again to handle is transferred to and is embathed in 0.01% dimethyl sulphoxide solution 2 times, each 15 minutes, removes residual cytochalasin B.
4. the Haliotis diversicolor gynogenesis diploid ovum after the above-mentioned processing is changed in the seawater,, produce Haliotis diversicolor gynogenesis diploid seedling by common Haliotis diversicolor seedling production method management.The chromosome counting result shows that staining of sperm body inactivation rate reaches 100%, and the dliploid inductivity reaches 95%～100%.The veliger survival rate reaches 96%～100%.
Embodiment 2
The Haliotis diversicolor gynogenetic haploid ovum that obtains by embodiment 1 method, after ovum starts 15 minutes, transfer in 300 μ mol/L 6-dimethylaminopurine (6-dimethylaminopurine) sea water solutions and handled 15 minutes, transfer in the seawater again, all the other get Haliotis diversicolor gynogenesis diploid seedling with the method for embodiment 1.
Claims (4)
1. the gynogenetic abductive approach of Haliotis diversicolor is characterized in that abductive approach is as follows:
1) genetic inactivation of sperm: with ultraviolet radiation Haliotis diversicolor seminal fluid, ultraviolet irradiating dose is 60000～120000 μ W/cm
2, seminal fluid thickness 1～2mm, sperm concentration 10
6～10
8Individual/ml;
2) the gynogenetic startup of ovum: the Haliotis diversicolor sperm of Haliotis diversicolor mature egg and process ultraviolet irradiation is mixed, and the control temperature is at 23～30 ℃, and the sperm ultimate density is no less than 10
5Individual/ml, the gynogenesis of startup Haliotis diversicolor ovum;
3) the gynogenetic haploid chromosome set doubles;
4) the Haliotis diversicolor gynogenesis diploid ovum after the above-mentioned processing is changed in the container for plant growth,, produce Haliotis diversicolor gynogenesis diploid seedling by common Haliotis diversicolor seedling production method management.
2, the gynogenetic abductive approach of Haliotis diversicolor as claimed in claim 1, it is characterized in that said gynogenetic haploid chromosome set doubles available cytochalasin B processing or handles with the 6-dimethylaminopurine in step 3), suppress to activate the 2nd reduction division of ovum.
3, the gynogenetic abductive approach of Haliotis diversicolor as claimed in claim 2, it is characterized in that in step 3) said gynogenetic haploid chromosome set doubles available cytochalasin B and handles and be: after ovum starts 7～20 minutes, transfer in the solution that contains 0.4～1.0mg/L cytochalasin B, make Haliotis diversicolor gynogenetic haploid chromosome set double to become gynogenesis diploid; Gynogenesis Haliotis diversicolor ovum after will doubling again to handle is transferred to and is embathed in 0.033%～0.05% dimethyl sulphoxide solution 2 times, each 10～15 minutes, removes residual cytochalasin B.
4, the gynogenetic abductive approach of Haliotis diversicolor as claimed in claim 1, it is characterized in that said the processing with the 6-dimethylaminopurine is in step 3): after ovum starts 7～20 minutes, transfer in the solution of 250～350 μ mol/L 6-dimethylaminopurines 10～20 minutes.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031329365A CN1201653C (en) | 2003-07-22 | 2003-07-22 | Induction method of gynogenesis of multicalor abalone |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB031329365A CN1201653C (en) | 2003-07-22 | 2003-07-22 | Induction method of gynogenesis of multicalor abalone |
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| Publication Number | Publication Date |
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| CN1476751A true CN1476751A (en) | 2004-02-25 |
| CN1201653C CN1201653C (en) | 2005-05-18 |
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| CNB031329365A Expired - Fee Related CN1201653C (en) | 2003-07-22 | 2003-07-22 | Induction method of gynogenesis of multicalor abalone |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100353923C (en) * | 2005-09-01 | 2007-12-12 | 厦门大学 | Gynogenesis inducing method for multicolor abalone |
| CN100387120C (en) * | 2006-06-08 | 2008-05-14 | 湖南师范大学 | A fish androgenesis method |
| CN101940183A (en) * | 2010-09-26 | 2011-01-12 | 浙江山下湖珍珠集团股份有限公司 | Method for cultivating pearl by inducing triploid hyriopsis cumingii |
| CN101569293B (en) * | 2009-05-31 | 2011-08-31 | 中国水产科学研究院东海水产研究所 | Method for inducing gynogenesis of Siberian sturgeon by using amur sturgeon sperm |
| CN103749362A (en) * | 2014-01-14 | 2014-04-30 | 扬州市嘉丰罗氏沼虾良种繁殖有限公司 | Macrobrachium rosenbergii breeding method |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100539832C (en) * | 2007-08-28 | 2009-09-16 | 中国水产科学研究院黄海水产研究所 | The method of inducing female nucleus growth with heterologous frozen sperm |
-
2003
- 2003-07-22 CN CNB031329365A patent/CN1201653C/en not_active Expired - Fee Related
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100353923C (en) * | 2005-09-01 | 2007-12-12 | 厦门大学 | Gynogenesis inducing method for multicolor abalone |
| CN100387120C (en) * | 2006-06-08 | 2008-05-14 | 湖南师范大学 | A fish androgenesis method |
| CN101569293B (en) * | 2009-05-31 | 2011-08-31 | 中国水产科学研究院东海水产研究所 | Method for inducing gynogenesis of Siberian sturgeon by using amur sturgeon sperm |
| CN101940183A (en) * | 2010-09-26 | 2011-01-12 | 浙江山下湖珍珠集团股份有限公司 | Method for cultivating pearl by inducing triploid hyriopsis cumingii |
| CN101940183B (en) * | 2010-09-26 | 2012-03-21 | 浙江山下湖珍珠集团股份有限公司 | Method for cultivating pearl by inducing triploid hyriopsis cumingii |
| CN103749362A (en) * | 2014-01-14 | 2014-04-30 | 扬州市嘉丰罗氏沼虾良种繁殖有限公司 | Macrobrachium rosenbergii breeding method |
| CN103749362B (en) * | 2014-01-14 | 2015-10-07 | 扬州市嘉丰罗氏沼虾良种繁殖有限公司 | A kind of Macrobrachium rosenbergii breeding method |
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| CN1201653C (en) | 2005-05-18 |
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