CN1317945C - Method for inducing stichopus japonicus triploid by static water pressure - Google Patents
Method for inducing stichopus japonicus triploid by static water pressure Download PDFInfo
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- CN1317945C CN1317945C CNB200410050418XA CN200410050418A CN1317945C CN 1317945 C CN1317945 C CN 1317945C CN B200410050418X A CNB200410050418X A CN B200410050418XA CN 200410050418 A CN200410050418 A CN 200410050418A CN 1317945 C CN1317945 C CN 1317945C
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Abstract
The present invention discloses a static water pressure method for inducing stichopus japonicus triploid, which is characterized in that the fertilized egg of stichopus japonicus is processed with the static water pressure method, the static water pressure is from 45 to 65Mpa, the starting time is from 35 to 65 min, and the duration time is from 3 to 6 min. The triploid stichopus japonicus is sterile normally or has low fecundity, the energy for breeding is converted into growing power, so the triploid stichopus japonicus has little aestivation time (even no aestivation time), high growth speed and good meat quality; simultaneously, the excellent variety variation and degradation caused by the miscellaneous variety of the stichopus japonicus in the process of breeding can be avoided, and the nutritive value of the excellent stichopus japonicus variety can be ensured. The static water pressure, the starting time and the duration time of the present invention can make the doubling rate, the hatching rate and the survival rate of the obtained fetuses or larvae enhanced, and the present invention provides a high-grade variety for the extensive breeding of the stichopus japonicus.
Description
Technical field:
The present invention relates to a kind of breeding method of stichopus japonicus new varieties, especially a kind of hydrostatic pressing is induced the triploid method of stichopus japonicus.
Background technology:
Stichopus japonicus belongs to Echinodermata, and it is coastal mainly to originate in ground such as China Liaoning, Shandong, and higher nutrition and and medical value are arranged.But in the 7～August in every year, stichopus japonicus is smart row, just stop to ingest after laying eggs, health shrinks hardening, thisly at high temperature do not eat motionless phenomenon and is " aestivation ", and aestivation is one of stichopus japonicus important physical activity.Between estivation period, internal organs such as stichopus japonicus digestive tract, respiratory trees are degenerated, and body weight obviously reduces, and the nutritive value loss is serious.Though at present people all have research to the utilization of stichopus japonicus nutritive value and cultural technique etc., on the public publication up to now also less than about how controlling the sterile report of stichopus japonicus.
Ploidy operation is an effective way of aquatile cell engineering breeding, since 1956 induce three quil fishes to obtain triploid first, has carried out the research of triploid induction on surplus surplus in the of the 20 kinds of fish, 30 kinds of shellfishes and 3 kinds of shell-fish.Up to the present, carrying out the ploidy method of operating mainly contains following several: a kind of is to handle fertilized egg with physical methods such as hot and cold shock and hydrostatic platen presses; Another kind is to handle the embryo who grows different times with chemical agent; Since stichopus japonicus be and " fish " class echinoderm far away of being separated by, the operation of existing ploidy directly can not be applied on the stichopus japonicus, to such an extent as to people can't be for aestivation time of reducing stichopus japonicus, improving its growth rate, to control stichopus japonicus artificially sterile.
Summary of the invention:
The present invention can not artificially control the sterile technical problem of stichopus japonicus in order to solve existing in prior technology, provides a kind of hydrostatic pressing to induce the triploid method of stichopus japonicus, can reduce effectively stichopus japonicus the aestivation time, improve its growth rate.
Technical solution of the present invention is: a kind of hydrostatic pressing is induced the triploid method of stichopus japonicus, it is characterized in that it being to handle stichopus japonicus fertilized egg with the hydrostatic platen press, and hydrostatic pressure is 45～65MPa, and zero-time is insemination back 35～65min, and the duration is 3～6min.
Described zero-time is insemination back 45～65min.
Described hydrostatic pressure is 55MPa, and zero-time is insemination back 55min, duration 5min.
Described fertilized egg can obtain as follows:
A. choose the stichopus japonicus in Chinese marine site or the stichopus japonicus in Russian Far East Japan marine site, artificial culture makes its sexual gland ripe simultaneously;
B. stimulate the stichopus japonicus parent participation with induced spawning method, induce the stichopus japonicus parent participation to lay eggs or arrange smart and collect the sperm or the ovum of the ovum of Chinese stichopus japonicus parent participation or sperm, the Japanese marine site stichopus japonicus parent participation in the Russian Far East respectively respectively;
C. carry out mating, generation fertilized egg with the sperm of the ovum of Chinese stichopus japonicus parent participation and the marine site stichopus japonicus parent participation of Russian Far East Japan or with the ovum of the sperm of Chinese stichopus japonicus parent participation and the Japanese marine site stichopus japonicus parent participation in the Russian Far East.
Described a step is for making Chinese stichopus japonicus parent participation gonad maturity hysteresis or the stichopus japonicus parent participation gonad maturity in Japanese marine site, the Russian Far East being shifted to an earlier date with artificial type of rearing.
Described Chinese stichopus japonicus parent participation gonad maturity is lagged behind be before Chinese stichopus japonicus parent participation gonad maturity 1～2 month and adopts catch parent participation and go into the pond, be no more than under 13～17 ℃ the condition constant temperature in temperature and cultivate to gonad maturity.
Described Russian Far East Japan marine site stichopus japonicus parent participation gonad maturity that makes is to adopt in 1～March of then 11～December or next year to catch parent participation and go into the pond in advance, heats up 1 ℃ every day, and until being warmed up to 15～16 ℃, constant temperature is cultivated, and accumulated temperature reaches 1200～1600 ℃ of skies.
Described b step is for drying in the shade the stimulation by running water method, and concrete grammar is with the stichopus japonicus parent participation 0.5～2h that dries in the shade, and uses stimulation by running water 0.5～1h again, then the pond that is added with fresh filtering sea is put in the stichopus japonicus parent participation.
Described b step is for the temperature difference method of hastening parturition, and concrete grammar is that the seawater that will store up the stichopus japonicus parent participation heats up or cooling heats up again or cooling, heat up to intersect and carry out, and the temperature difference is 2～5 ℃, until laying eggs or arrange essence.
Described b step is the method for hastening parturition with the sea water immersion of process ultraviolet irradiation, concrete grammar is that elder generation is 30～100 watts quartz burner irradiation seawater with power, exposure dose is 100～200mwh/l, afterwards the stichopus japonicus parent participation is inserted wherein and soaks, until laying eggs or arranging smart.
The present invention handles stichopus japonicus fertilized egg with the hydrostatic platen press, makes stichopus japonicus fertilized ovum chromosome polyploidization.The triploid stichopus japonicus is generally sterile or fertility is low, the energy that is used to breed is converted into the growing power of self, triploid stichopus japonicus aestivation time few (even not having the aestivation time) for this reason, fast growth and meat are good, the improved seeds variation that can avoid simultaneously stichopus japonicus germplasm in the breeding process to mix and cause and degenerating has guaranteed the nutritive value of improved seeds stichopus japonicus.Hydrostatic pressure of the present invention, initial and duration can make the embryo of gained and doubly change rate of larva, incubation rate and survival rate higher, and the kind of high-quality is provided for the large-scale farming of stichopus japonicus.Especially use the stichopus japonicus in Chinese marine site or the stichopus japonicus in Russian Far East Japan marine site, artificial culture makes its sexual gland ripe simultaneously, stimulate the stichopus japonicus parent participation with induced spawning method again, induce the stichopus japonicus parent participation to lay eggs or arrange smart and collect the ovum or the sperm of Chinese stichopus japonicus parent participation respectively respectively, the sperm or the ovum of the marine site stichopus japonicus parent participation of Russia Far East Japan, carry out fertilized egg that mating produces with the sperm of the ovum of Chinese stichopus japonicus parent participation and the marine site stichopus japonicus parent participation of Russian Far East Japan or with the ovum of the sperm of Chinese stichopus japonicus parent participation and the Japanese marine site stichopus japonicus parent participation in the Russian Far East then, its fertilized egg survival rate height and can improve the quality of stichopus japonicus.
Embodiment:
Embodiment 1:
A. use the sperm ovum combination of Chinese marine site stichopus japonicus, generate fertilized egg;
B. the fertilized egg with zero-time (insemination back) 35～65min places the pressure storehouse, water temperature is for being fit to the temperature of fertilized egg survival, be preferably 18～23 ℃, handle fertilized egg with the hydrostatic platen press, hydrostatic pressure 45～65MPa, duration 3～6min then makes the diplodization of stichopus japonicus fertilized ovum chromosome, tripling, four times of changes.
Embodiment 2:
A. use the stichopus japonicus in Chinese marine site or the stichopus japonicus in Russian Far East Japan marine site, artificial culture makes its sexual gland ripe simultaneously;
B. stimulate the stichopus japonicus parent participation with induced spawning method, induce the stichopus japonicus parent participation to lay eggs or arrange smart and collect the sperm or the ovum of the ovum of Chinese stichopus japonicus parent participation or sperm, the Japanese marine site stichopus japonicus parent participation in the Russian Far East respectively respectively;
C. carry out mating, generation fertilized egg with the sperm of the ovum of Chinese stichopus japonicus parent participation and the marine site stichopus japonicus parent participation of Russian Far East Japan or with the ovum of the sperm of Chinese stichopus japonicus parent participation and the Japanese marine site stichopus japonicus parent participation in the Russian Far East;
D. the fertilized egg with zero-time (insemination back) 45～65min places the pressure storehouse, water temperature is for being fit to the temperature of fertilized egg survival, be preferably 18～23 ℃, handle fertilized egg with the hydrostatic platen press, hydrostatic pressure 45～65MPa, duration 3～6min then makes the diplodization of stichopus japonicus fertilized ovum chromosome, tripling, four times of changes.
Embodiment 3:
A. be used in preceding 1～2 month of Chinese stichopus japonicus parent participation sexual gland, be i.e. in 3～May then, adopt the thorn of catching Chinese marine site and participate in the pond, be no more than constant temperature cultivation under 13～17 ℃ of conditions in temperature.The constant temperature nurturing period can be mixed the mud liquid feeding that organic silt forms by seawood meal, also can drop into and contain mussel powder, shrimp med, soya-bean milk etc. at interior synthetic diet, until gonad maturity.At this moment, the gonad maturity phase of the Chinese stichopus japonicus of artificial culture lags behind than the Chinese stichopus japonicus in the nature marine site, generally is 8～September in every year, and is ripe simultaneously with the sexual gland of marine site, the Russian Far East (Korea peninsula) stichopus japonicus of normal growth.
B. will store up seawater in the holding pond of Chinese stichopus japonicus parent participation, Russian Far East Japan marine site stichopus japonicus parent participation and put driedly, generally be to carry out after between the lights, allows the stichopus japonicus parent participation dry in the shade 45 minutes to 1 hour.Use water slug then,,, then needed to impact 40～45 minutes as impacting with sluggish flow as then impacting 10～15 minutes with water under high pressure.Afterwards the pond that is added with fresh filtering sea is put in Chinese stichopus japonicus parent participation, the marine site stichopus japonicus parent participation of Russian Far East Japan respectively.Generally speaking about 2 hours, the stichopus japonicus parent participation begins to creep to the top layer pool wall, movable frequent, head lifted swing, the male row's essence that begins, the female beginning lays eggs after half an hour, collects the ovum or the sperm of Chinese stichopus japonicus parent participation respectively, the sperm or the ovum of the Japanese marine site stichopus japonicus parent participation in the Russian Far East.Promptly hasten parturition with the flowing water method of drying in the shade.
C. hybridize generation fertilized egg with the ovum of the Japanese marine site stichopus japonicus parent participation in the Russian Far East with the sperm of the ovum of Chinese stichopus japonicus parent participation and the marine site stichopus japonicus parent participation of Russian Far East Japan or with the sperm of Chinese stichopus japonicus parent participation;
D. the fertilized egg with zero-time (insemination back) 55min places the pressure storehouse, water temperature is for being fit to the temperature of fertilized egg survival, be preferably 18～23 ℃, handle fertilized egg with the hydrostatic platen press, hydrostatic pressure 55MPa, duration 5min then makes the diplodization of stichopus japonicus fertilized ovum chromosome, tripling, four times of changes.
Embodiment 4:
A. adopt in the January of then December or next year and catch the thorn of choosing marine site, the Russian Far East (Korea peninsula) and participate in the pond, heat up 1 ℃ every day, until being warmed up to 15～16 ℃, constant temperature is cultivated, the constant temperature nurturing period can be mixed the mud liquid feeding that organic silt forms by seawood meal, also can drop into and contain mussel powder, shrimp med, soya-bean milk etc. at interior synthetic diet.Accumulated temperature reaches 1200～1600 ℃ of skies, gonad maturity, at this moment, the gonad maturity phase of the Japanese marine site stichopus japonicus parent participation in the Russian Far East of artificial culture than Japanese marine site, the Russian Far East stichopus japonicus in the nature marine site in advance, generally be 5～July in every year, ripe simultaneously with the sexual gland of the Chinese stichopus japonicus of normal growth.Can select the stichopus japonicus parent participation of zones of different in the Chinese marine site according to the Russian Far East Japanese marine site stichopus japonicus parent participation gonad maturity degree, adapt with gonad maturity degree with the Japanese marine site stichopus japonicus parent participation in the Russian Far East.
The cultivation ocean temperature of storing up the stichopus japonicus parent participation is changed, and the temperature difference is 2～5 ℃.The stichopus japonicus parent directly can be inserted in the high water temperature (directly heating up), or after the stichopus japonicus parent participation handled with low water temperature earlier, handle (lower the temperature earlier and afterwards heat up) with high water temperature again; Or with the stichopus japonicus parent participation with height water temperature repeated treatments (high low temperature interactive operation), the processing time that once rises (falling) water temperature usually is 1～2 hour, need decide on stichopus japonicus parent participation situation.If find to have (row the is smart) sign of laying eggs, should shift out individually in normal water temperature environment, wait to lay eggs or arrange smartly, collect the ovum or the sperm of Chinese stichopus japonicus parent participation respectively, the sperm or the ovum of the Japanese marine site stichopus japonicus parent participation in the Russian Far East.Promptly hasten parturition with temperature differential method.
C. hybridize generation fertilized egg with the ovum of the Japanese marine site stichopus japonicus parent participation in the Russian Far East with the sperm of the ovum of Chinese stichopus japonicus parent participation and the marine site stichopus japonicus parent participation of Russian Far East Japan or with the sperm of Chinese stichopus japonicus parent participation.The quantity of sperm generally is controlled at 100～10000 around each ovum.
C. the fertilized egg with zero-time (insemination back) 45～65min places the pressure storehouse, water temperature is for being fit to the temperature of fertilized egg survival, be preferably 18-23 ℃, handle fertilized egg with the hydrostatic platen press, hydrostatic pressure 45～65MPa, duration 3～6min then makes the diplodization of stichopus japonicus fertilized ovum chromosome, tripling, four times of changes.
Embodiment 5:
A. the method with embodiment 3 or embodiment 4 makes its sexual gland ripe simultaneously;
B. hasten parturition with the ultraviolet sea water immersion method of hastening parturition.Concrete grammar is earlier to be 30～100 watts quartz burner irradiation seawater with power, and exposure dose is 100～200mwh/l, afterwards the stichopus japonicus parent participation is inserted wherein and soaks, until laying eggs or arranging essence; Seawater can be static or mobile, if circulating water, flow rate can be 10 liters/minute, and circulation irradiation 1 hour is soaked the stichopus japonicus parent participation afterwards to insert wherein, until laying eggs or arranging essence, move in the normal seawater.Preferably in ultraviolet ray circulation irradiation seawater, do abundant aeration, make gas meltage abundance in the water.After waiting to lay eggs or arranging essence, collect the ovum or the sperm of Chinese stichopus japonicus parent participation respectively, the sperm or the ovum of the marine site stichopus japonicus parent participation of Russian Far East Japan.
Other step is also with embodiment 3 or 4.
Induced spawning method can use the disclosed method of above embodiment separately, also it can be merged and use.If the stichopus japonicus parent participation can be told male and female, then can be respectively in groups female, male sea cucumber be carried out artificial induced spawning, otherwise, then should carry out artificial induced spawning to the single individuality of stichopus japonicus parent participation respectively.
Experimental result sees Table 1, table 2.
Table 1 different pressures, duration are handled stichopus japonicus embryo's times change rate down
| Experiment condition | Fertilization rate (%) | Incubation rate (%) | Times change rate | ||||
| Pressure (MPa) | Zero-time (min) | Duration (min) | 2n(％) | 3n(％) | 4n(％) | ||
| Contrast | - | - | 99.0 | 88.0 | 100 | - | - |
| 45 | 55 | 3 | 98.0 | 30.0 | 54.8 | 16.6 | 6.9 |
| 45 | 55 | 6 | 98.0 | 12.0 | 7.2 | 8.4 | 41.3 |
| 55 | 55 | 3 | 99.0 | 15.0 | 11.1 | 6.3 | 34.3 |
| 55 | 55 | 6 | 98.0 | 7.0 | 6.3 | 8.4 | 75.0 |
| 65 | 55 | 3 | 99.0 | 4.0 | 9.7 | 5.3 | 38.4 |
| 65 | 55 | 6 | 98.0 | 1.0 | 7.0 | 6.8 | 46.9 |
The different zero-times of table 2 are handled stichopus japonicus embryo's times change rate down
| Experiment condition | Fertilization rate (%) | Incubation rate (%) | Times change rate | ||||
| Pressure (MPa) | Zero-time (min) | Duration (min) | 2n(％) | 3n(％) | 4n(％) | ||
| Contrast | - | - | 99.0 | 85.0 | 100 | - | - |
| 55 | 40 | 5 | 94.0 | 50.0 | 87.4 | 9.5 | 4.2 |
| 55 | 45 | 5 | 99.0 | 45.0 | 75.7 | 13.4 | 10.9 |
| 55 | 50 | 5 | 95.0 | 50.0 | 67.3 | 17.3 | 15.4 |
| 55 | 55 | 5 | 99.0 | 10.0 | 49.1 | 15.1 | 35.8 |
| 55 | 60 | 5 | 95.0 | 4.0 | 34.0 | 11.8 | 39.1 |
| 55 | 65 | 5 | 99.0 | 1.0 | 62.6 | 8.9 | 14.3 |
Annotate: 2n, 3n, 4n represent two, three respectively, tetraploid
Claims (10)
1. a hydrostatic pressing is induced the triploid method of stichopus japonicus, it is characterized in that it being to handle stichopus japonicus fertilized egg with the hydrostatic platen press, and hydrostatic pressure is 45～65MPa, and zero-time is insemination back 35～65min, and the duration is 3～6min.
2. hydrostatic pressing according to claim 1 is induced the triploid method of stichopus japonicus, it is characterized in that: described zero-time is insemination back 45～65min.
3. hydrostatic pressing according to claim 1 and 2 is induced the triploid method of stichopus japonicus, it is characterized in that: described hydrostatic pressure is 55MPa, and zero-time is insemination back 55min, duration 5min.
4. hydrostatic pressing according to claim 1 and 2 is induced the triploid method of stichopus japonicus, it is characterized in that described fertilized egg obtains as follows:
A. choose the stichopus japonicus in Chinese marine site or the stichopus japonicus in Russian Far East Japan marine site, artificial culture makes its sexual gland ripe simultaneously;
B. stimulate the stichopus japonicus parent participation with induced spawning method, induce the stichopus japonicus parent participation to lay eggs or arrange smart and collect the sperm or the ovum of the ovum of Chinese stichopus japonicus parent participation or sperm, the Japanese marine site stichopus japonicus parent participation in the Russian Far East respectively respectively;
C. carry out mating, generation fertilized egg with the sperm of the ovum of Chinese stichopus japonicus parent participation and the marine site stichopus japonicus parent participation of Russian Far East Japan or with the ovum of the sperm of Chinese stichopus japonicus parent participation and the Japanese marine site stichopus japonicus parent participation in the Russian Far East.
5. hydrostatic pressing according to claim 4 is induced the triploid method of stichopus japonicus, it is characterized in that: described a step is for making Chinese stichopus japonicus parent participation gonad maturity hysteresis or the stichopus japonicus parent participation gonad maturity in Japanese marine site, the Russian Far East being shifted to an earlier date with artificial type of rearing.
6. hydrostatic pressing according to claim 5 is induced the triploid method of stichopus japonicus, it is characterized in that: described Chinese stichopus japonicus parent participation gonad maturity is lagged behind be before Chinese stichopus japonicus parent participation gonad maturity 1～2 month and adopts catch parent participation and go into the pond, be no more than under 13～17 ℃ the condition constant temperature in temperature and cultivate to gonad maturity.
7. hydrostatic pressing according to claim 5 is induced the triploid method of stichopus japonicus, it is characterized in that: described Russian Far East Japan marine site stichopus japonicus parent participation gonad maturity that makes is to adopt in 1～March of then 11～December or next year to catch parent participation and go into the pond in advance, heat up 1 ℃ every day, until being warmed up to 15～16 ℃, constant temperature is cultivated, and accumulated temperature reaches 1200～1600 ℃ of skies.
8. hydrostatic pressing according to claim 4 is induced the triploid method of stichopus japonicus, it is characterized in that: described b step is with drying in the shade the stimulation by running water method, concrete grammar is with the stichopus japonicus parent participation 0.5～2h that dries in the shade, use stimulation by running water 0.5～1h again, then the pond that is added with fresh filtering sea is put in the stichopus japonicus parent participation.
9. hydrostatic pressing according to claim 4 is induced the triploid method of stichopus japonicus, it is characterized in that: described b step is the method for hastening parturition with temperature difference, concrete grammar be will store up the seawater of stichopus japonicus parent participation heat up or cooling heats up again or cooling, heat up to intersect and carry out, the temperature difference is 2～5 ℃, until laying eggs or arranging smart.
10. hydrostatic pressing according to claim 4 is induced the triploid method of stichopus japonicus, it is characterized in that: described b step is the method for hastening parturition with the sea water immersion of process ultraviolet irradiation, concrete grammar is that elder generation is 30～100 watts quartz burner irradiation seawater with power, exposure dose is 100～200mwh/l, afterwards the stichopus japonicus parent participation is inserted wherein and soak, until laying eggs or arranging smart.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101940178A (en) * | 2010-08-31 | 2011-01-12 | 厦门大学 | Method for improving survival rate of oysters by high static water pressure treatment |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102100198A (en) * | 2010-12-20 | 2011-06-22 | 山东海益宝水产股份有限公司 | Method for producing triploid stichopus japonicus seedling |
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| US4834024A (en) * | 1984-09-06 | 1989-05-30 | Board Of Regents Of The University Of Washington | Inducing polyploidy in oysters |
| CN1340297A (en) * | 2000-08-25 | 2002-03-20 | 中国水产科学研究院黄海水产研究所 | Method for producing triploid of young Jiekong scallop |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4834024A (en) * | 1984-09-06 | 1989-05-30 | Board Of Regents Of The University Of Washington | Inducing polyploidy in oysters |
| CN1340297A (en) * | 2000-08-25 | 2002-03-20 | 中国水产科学研究院黄海水产研究所 | Method for producing triploid of young Jiekong scallop |
Non-Patent Citations (3)
| Title |
|---|
| 仿刺参(Apostichopus japonicus)多倍体诱导的初步研究 常亚青等，大连水产学院学报，第17卷第1期 2002 * |
| 仿刺参(Apostichopus japonicus)多倍体诱导的初步研究 常亚青等，大连水产学院学报，第17卷第1期 2002;静水压休克诱导近江牡蛎三倍体的发生 陈晓汉等，广西科学，第1卷第1期 1994 * |
| 静水压休克诱导近江牡蛎三倍体的发生 陈晓汉等，广西科学，第1卷第1期 1994 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101940178A (en) * | 2010-08-31 | 2011-01-12 | 厦门大学 | Method for improving survival rate of oysters by high static water pressure treatment |
| CN101940178B (en) * | 2010-08-31 | 2012-05-09 | 厦门大学 | Method for improving survival rate of oysters by high static water pressure treatment |
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| CN1586190A (en) | 2005-03-02 |
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