CN1201653C - Induction method of gynogenesis of multicalor abalone - Google Patents
Induction method of gynogenesis of multicalor abalone Download PDFInfo
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- CN1201653C CN1201653C CNB031329365A CN03132936A CN1201653C CN 1201653 C CN1201653 C CN 1201653C CN B031329365 A CNB031329365 A CN B031329365A CN 03132936 A CN03132936 A CN 03132936A CN 1201653 C CN1201653 C CN 1201653C
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- gynogenesis
- ovum
- haliotis diversicolor
- abalone
- multicolor
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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
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Abstract
The present invention relates to an induction method of gynogenesis of a multicolor abalone, which comprises the following steps: firstly, multicolor abalone semen is radiated by ultraviolet rays, and the genetic materials of the multicolor abalone sperms are deactivated; subsequently, the concentration of the genetic deactivated sperms is adjusted, and then, the genetic deactivated sperms are mixed with the ova of the multicolor abalone; the gynogenesis of the ova of the multicolor abalone is carried out, and then, the cytochalasin B or 6-dimethylaminopurine is used for inhibiting and activating the maiosis of the ova of the second time to cause the genome to be doubled to generate gynogenesis diploid; the diploid ova of the gynogenesis of the multicolor abalone, which are treated, are transferred into a cultivating container to be managed according to a common multicolor abalone cultivating and producing method, and then, the diploid culture of the gynogenesis of the multicolor abalone is produced. The survival rate of the filial generation which is induced by the method of the present invention is high, the survival rate reaches 99.5% in the veliger stage, which is farther greater than the survival rate (3.3% to 9.0%) of a larva in the same stage by using the Fujino induction method, etc. for haliotis discus hannai ino; in addition, the diploid inductivity reaches 95% to 100%, and the ovum starting rate reaches 96% to 100%, which is larger than the ovum starting rate (36% to 49%) in the gynogenesis of the haliotis discus hannai ino in the report of the Arai, etc.
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 (3)
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, and said gynogenetic haploid chromosome set doubles to handle or handle with the 6-dimethylaminopurine with cytochalasin B, suppresses to activate the 2nd reduction division of ovum.
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 the management of Haliotis diversicolor seedling production method.
2, the gynogenetic abductive approach of Haliotis diversicolor as claimed in claim 1, it is characterized in that in step 3) that said gynogenetic haploid chromosome set doubles to handle with cytochalasin B is: 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.
3, 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 |
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| CNB031329365A CN1201653C (en) | 2003-07-22 | 2003-07-22 | Induction method of gynogenesis of multicalor abalone |
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| CNB031329365A CN1201653C (en) | 2003-07-22 | 2003-07-22 | Induction method of gynogenesis of multicalor abalone |
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| CN1476751A CN1476751A (en) | 2004-02-25 |
| CN1201653C true CN1201653C (en) | 2005-05-18 |
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Cited By (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 |
Families Citing this family (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 |
| 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 |
| 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 |
| 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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2003
- 2003-07-22 CN CNB031329365A patent/CN1201653C/en not_active Expired - Fee Related
Cited By (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 |
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