CN103202214A - Breeding method of sargassum vachellianum artificial seeds - Google Patents
Breeding method of sargassum vachellianum artificial seeds Download PDFInfo
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- CN103202214A CN103202214A CN2013101487759A CN201310148775A CN103202214A CN 103202214 A CN103202214 A CN 103202214A CN 2013101487759 A CN2013101487759 A CN 2013101487759A CN 201310148775 A CN201310148775 A CN 201310148775A CN 103202214 A CN103202214 A CN 103202214A
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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
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Abstract
The invention relates to a breeding method of algae and particularly relates to a breeding method of sargassum vachellianum artificial seeds. The breeding method comprises the following steps: temporarily culturing adult algae of sargassum vachellianum in a culture solution for 18-22 days under the condition that the temperature is 16-20 DEG C, and replacing one half of the culture solution every 5-8 days till the appearance of female and male reproductive organs; charging gas for culturing for 1-5 days according to the proportion that the quantity of the female reproductive organs to the quantity of the male reproductive organs is 1: 1, and replacing one half of the culture solution every 1-3 days till fertilized eggs begin to fall off and settle; and collecting the fertilized eggs which begin to fall off and settle, standing and culturing for 18-25 days under the conditions that the temperature is 21-24 DEG C, the light intensity is 1800-2200lx and the lighting cycle is 14L: 10D. According to the method, the sexual reproduction time can be effectively controlled and prolonged, the higher fertilization rate and the survival rate of seedlings can be ensured, and the fertilization rate can be above 95% and are not affected by natural conditions and climate changes.
Description
Technical field
The present invention relates to the mating system of a kind of algae, particularly a kind of Wa Shi sargassum artificial seed's mating system.
Background technology
Sargassum Phaeophyta, Sargassaceae.China is one of main place of production of sargassum, and kind more than 60 is arranged, and common have decomposite leaf sargassum, half leaf sargassum, flagellum sargassum, Heng Shi sargassum, Sargassum, a Wa Shi sargassum etc.Wherein, the Wa Shi sargassum, Latin is called Sargassum vachellianum Greville, be that a kind of concentrated ground such as China Zhejiang, Fujian, Guangdong and Hong Kong that are distributed in are coastal, Wa Shi sargassum itself is widely applied to medicine, food, feed and organic manure field because being rich in protein, carbohydrate, mineral salt, vitamin etc.Especially the organic manure field quite has been subjected to people's attention.Wa Shi sargassum fertilizer is compared chemical fertilizer, and natural advantage is arranged, and does not have secondary pollution, and for crop and soil without any harm, the growth of plant is played significant adjusting and facilitation.
Wa Shi sargassum plant is tall and big, and is with luxuriant foliage and spreading branches in leafy profusion, is grown on the cay of subtidal zone shallow sea more, floats on the water surface in flakes, is one of important member of algae field, east China seabed.The fragile marine eco-environment just as " pasture, seabed ", can be repaired in algae field, seabed.In recent years, marine environment worsens, and the eutrophication degree of marine site seawater is than higher, and the content height of nitrogen, P elements causes harm such as red tide easily, halobiontic growth has been caused have a strong impact on.And the sea jungle of being formed by algae such as Wa Shi sargassums, energy purifying water body environment, prevent the destruction of marine environment, also be the halobiontic life ground of perching of multiple class, the desirable place of having kept away enemy, forage, having laid eggs for they provide, life is flourishing, the ecosystem of steady change thereby form.
The quick growth of Wa Shi sargassum and huge biomass cause ocean worker's concern and interest, are classified as by the marine ecology scholar and rebuild algae field, seabed and implement one of important species of marine ecology reparation.But almost do not study about reproductive biology and the offspring breeding method of such marine alga at present, so, carry out the indoors artificial accelerating and become algae, finish sexual reproduction, it is significant that acquisition fertilized egg is carried out the artificial seedling rearing.
Summary of the invention
The mating system that the purpose of this invention is to provide a kind of Wa Shi sargassum artificial seed can realize that female, male receptacle is ripe synchronously, improves survival rate and emergence rate.
Purpose of the present invention can be achieved through the following technical solutions:
A kind of Wa Shi sargassum artificial seed's mating system, its step comprises:
(1) the Wa Shi sargassum that will grow panicle shape receptacle blank is as becoming algae, in temperature is under 16-20 ℃ the condition, to support 18-22 days temporarily in culture fluid, changes half culture fluid in every 5-8 days, till female, male receptacle occurs;
(2) get the one-tenth algae side shoot that has receptacle in the step (1), according to the ratio of female, male receptacle quantity 1:1, inflation was cultivated 1-5 days, changed half culture fluid in every 1-3 days, till fertilized egg beginning comes off sedimentation; The inflation cultivation temperature is 21-24 ℃, intensity of illumination 1500-2300lx, periodicity of illumination 14L:10D;
(3) collecting the fertilized egg of sedimentation that comes off that begins in the step (2), is 21-24 ℃ in temperature, and intensity of illumination is 1800-2200lx, under the condition of periodicity of illumination 14L:10D, leaves standstill and cultivates 18-25 days, can breed into seedling;
The condition of supporting temporarily in the described step (1) is intensity of illumination 1500-2000lx, periodicity of illumination 10L:14D.
In the described step (2), becoming the length of algae side shoot is 2-6cm, a tool 15-20 receptacle on this side shoot.
In the described step (3), the fertilized egg when collection begins to come off sedimentation 10-14h from becoming the algae side shoot is cultivated, and the culture density of this fertilized egg is 20-80/cm
2Preferably, the fertilized egg when collection begins to come off sedimentation 12h from becoming the algae side shoot is cultivated, and the culture density of this fertilized egg is 40-50/cm
2
In the described step (3), the length of seedling is 3-6mm, has 2-4 algae leaf.
The seedling inflation that obtains in the step (3) was cultivated 10-30 days, changed half culture fluid in every 3-6 days, cultivation temperature is 20-26 ℃, and intensity of illumination is 3000-3500lx, and periodicity of illumination is 14L:10D.Purpose is in order to accelerate growth of seedlings, to carry out sea propagation and artificial cultivation better.
Described culture fluid is the MES medium with the sterilization seawater is the mixed liquor that the ratio of 1:0.01-0.03 mixes according to volume ratio; Also contain the KI that concentration is 0.5-1.0mg/L in the described culture fluid.
The component of described MES medium is EDTA2Na, H
3BO
3, MnCl
24H
2O, FeCl
36H
2O, ZnCl
2, CoCl
26H
2O, ironic citrate, NaNO
3, sodium glycero-phosphate, trishydroxymethylaminomethane, the respective concentration of each component is 0.3-0.8g/L, 0.4-0.6g/L, 0.05-0.09g/L, 0.01-0.03g/L, 0.002-0.007g/L, 0.001-0.003g/L, 0.01-0.03g/L, 1-4g/L, 0.2-0.6g/L, 2-7g/L, and the pH value of this MES medium is 7-8.
Compared with prior art, beneficial effect of the present invention is:
1, the inventive method is under the manual control condition, support the also male and female parent algae of accelerating Wa Shi sargassum temporarily, ovum and sperm in the sexual reproduction process are diffused synchronously, finishing gamete engages, not only can control effectively and prolong the sexual propagation time, and can guarantee higher fertilization rate and seedling percent, more than the fertilization rate reached to 95%.
2, the inventive method is not subjected to the influence of natural conditions, climatic variation, in the relatively long time period, can collect seedling and grow seedlings as required by being controlled to the maturity of algae, obtains artificial seed in batch, is used for marine propagation or artificial cultivation.
3, the inventive method adopts inflation to cultivate to obtain to transfer to after the fertilized egg and leaves standstill cultivation, has avoided the mutual adhesion of Wa Shi sargassum juvenile sporophyte rhizoid, has guaranteed the normal growth of seedling.
Embodiment
Below in conjunction with embodiment, the invention will be further described:
The preparation of culture fluid:
With 3.0g EDTA2Na, 2.5g H
3BO
3, 0.35g MnCl
24H
2O, 2mL FeCl
36H
2O solution (62.5mg/mL), 2mL ZnCl
2Solution (12.5mg/mL), 2mL CoCl
26H
2O solution (5mg/mL), 2mL ironic citrate solution (30mg/mL) mix, and are settled to 1000mL with distilled water, are made into MPII solution and then get 200mL MP II solution and 2.8g NaNO
3, 0.4g sodium glycero-phosphate, 4.0g trishydroxymethylaminomethane and 700mL distilled water mixes, and is settled to 1000mL, is made into the MES medium, the pH value of this MES medium is 7.6, will be stored in refrigerator behind the MES medium autoclaving then;
The MES medium that adds 20mL in 1L sterilization seawater is formed the culture fluid of Wa Shi sargassum, in order to provide the Wa Shi sargassum required nutritive element iodine better, adds KI again in culture fluid, makes that the KI concentration in the culture fluid is 1mg/L.
Embodiment
(1) selection of one-tenth algae:
Collection grows in the natural waters subtidal zone, has grown panicle shape receptacle blank, still can't distinguish the male and female individuality, and epizoite is less, and the Wa Shi sargassum that is in a good state of health is as becoming algae.
(2) become supporting temporarily of algae:
Become algae to support temporarily in the cultivation bucket that is placed with culture fluid with the sterilization seawater, changed half seawater, and supported 20 days temporarily in per 7 days.Support condition temporarily: 18 ℃ of temperature, intensity of illumination 1500-2000lx, periodicity of illumination 10L:14D.
(3) receptacle accelerating:
The one-tenth algae of supporting temporarily in the step (2) is cut the side shoot with receptacle, the long 3-5cm of each side shoot, have 15-20 receptacle on it, ratio according to female, male receptacle 1:1, inflation is incubated in the same gas filling bottle (filling air with air pump), inflation was cultivated 2-3 days, changed half culture fluid every day.Condition of culture: temperature 21-24 ℃, intensity of illumination 2000lx, periodicity of illumination 14L:10D.
(4) cultivation of seedling:
Collect fertilized egg when fertilized egg beginning comes off after the sedimentation 12h, this moment, fertilized egg was just grown to the juvenile sporophyte that grows 4-8 rhizoid bud, can adhesion not become piece each other, easily realized evenly disperseing, leave standstill and cultivated 20 days, the culture density of fertilized egg be 40-50 individual/cm
2, change half culture fluid every day, condition of culture, temperature 21-24 ℃, intensity of illumination 2000lx, periodicity of illumination 14L:10D can finish the cultivation of seedling.
In order to accelerate growth of seedlings, can carry out sea propagation and artificial cultivation better, above-mentioned seedling is further inflated cultivation, concrete steps are, with 20 days seedling of static cultivation, the about 5mm of length, have 2-4 algae leaf, inflation was cultivated 20 days, changed half culture fluid in per 5 days.Condition of culture: temperature 21-24 ℃, intensity of illumination 3000-3500lx, periodicity of illumination 14L:10D.
The fertilization rate of the fertilized egg of the Wa Shi sargassum in the present embodiment has improved the breeding rate of Wa Shi sargassum greatly more than 95%.
Because the rhizoid viscosity that Wa Shi sargassum juvenile sporophyte grows is very big, so in the present embodiment, adopt the inflation best cultivation to cultivate the fertilized egg of obtaining earlier, and then transfer to and leave standstill cultivation, the juvenile sporophyte that obtains has been avoided the mutual adhesion of rhizoid, guarantee the normal growth of seedling, improved emergence rate.In the prior art, adopt the inflation best cultivation to breed into seedling fertilized egg, without the step that leaves standstill cultivation, cause the agglomerating phenomenon of the mutual adhesion of rhizoid of juvenile sporophyte to take place, emergence rate obviously lowers.
The above is preferred embodiment of the present invention, but the present invention should not be confined to the disclosed content of this embodiment.So everyly do not break away from the equivalence of finishing under the spirit disclosed in this invention or revise, all fall into the scope of protection of the invention.
Claims (9)
1. a Wa Shi sargassum artificial seed mating system, its step comprises:
(1) the Wa Shi sargassum that will grow panicle shape receptacle blank is as becoming algae, in temperature is under 16-20 ℃ the condition, to support 18-22 days temporarily in culture fluid, changes half culture fluid in every 5-8 days, till female, male receptacle occurs;
(2) get the one-tenth algae side shoot that has receptacle in the step (1), according to the ratio of female, male receptacle quantity 1:1, inflation was cultivated 1-5 days, changed half culture fluid in every 1-3 days, till fertilized egg beginning comes off sedimentation; The inflation cultivation temperature is 21-24 ℃, intensity of illumination 1500-2300lx, periodicity of illumination 14L:10D;
(3) collecting the fertilized egg of sedimentation that comes off that begins in the step (2), is 21-24 ℃ in temperature, and intensity of illumination is 1800-2200lx, under the condition of periodicity of illumination 14L:10D, leaves standstill and cultivates 18-25 days, can breed into seedling.
2. Wa Shi sargassum artificial seed's according to claim 1 mating system is characterized in that: the condition of supporting temporarily in the described step (1) is intensity of illumination 1500-2000lx, periodicity of illumination 10L:14D.
3. Wa Shi sargassum artificial seed's according to claim 1 mating system is characterized in that: in the described step (2), becoming the length of algae side shoot is 2-6cm, a tool 15-20 receptacle on this side shoot.
4. Wa Shi sargassum artificial seed's according to claim 1 mating system, it is characterized in that: in the described step (3), fertilized egg when collection begins to come off sedimentation 10-14h from becoming the algae side shoot is cultivated, and the culture density of this fertilized egg is 20-80/cm
2
5. Wa Shi sargassum artificial seed's according to claim 4 mating system is characterized in that: in the described step (3), the fertilized egg of collecting when becoming the algae side shoot to begin to come off sedimentation 12h is cultivated, and the culture density of this fertilized egg is 40-50/cm
2
6. Wa Shi sargassum artificial seed's according to claim 1 mating system, it is characterized in that: in the described step (3), the length of seedling is 3-6mm, has 2-4 algae leaf.
7. Wa Shi sargassum artificial seed's according to claim 1 mating system, it is characterized in that: the seedling inflation that step (3) obtains was cultivated 10-30 days, changed half culture fluid in every 3-6 days, cultivation temperature is 20-26 ℃, intensity of illumination is 3000-3500lx, and periodicity of illumination is 14L:10D.
8. according to claim 1 or 7 described Wa Shi sargassum artificial seeds' mating system, it is characterized in that: described culture fluid is the MES medium with the sterilization seawater is the mixed liquor that the ratio of 1:0.01-0.03 mixes according to volume ratio; Also contain the KI that concentration is 0.5-1.0mg/L in the described culture fluid.
9. Wa Shi sargassum artificial seed's according to claim 8 mating system, it is characterized in that: the component of described MES medium is EDTA2Na, H
3BO
3, MnCl
24H
2O, FeCl
36H
2O, ZnCl
2, CoCl
26H
2O, ironic citrate, NaNO
3, sodium glycero-phosphate, trishydroxymethylaminomethane, the respective concentration of each component is 0.3-0.8g/L, 0.4-0.6g/L, 0.05-0.09g/L, 0.01-0.03g/L, 0.002-0.007g/L, 0.001-0.003g/L, 0.01-0.03g/L, 1-4g/L, 0.2-0.6g/L, 2-7g/L, and the pH value of this MES medium is 7-8.
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Cited By (9)
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| CN103651118A (en) * | 2013-11-11 | 2014-03-26 | 青岛文创科技有限公司 | Method for sterilizing turbinaria conoides rhizoid explant |
| CN104115738A (en) * | 2014-07-29 | 2014-10-29 | 广东海洋大学 | Seedling method for inducing synchronous maturity of Leizhou gulfweed in vitro receptacles |
| CN105393909A (en) * | 2015-11-25 | 2016-03-16 | 上海海洋大学 | Sargassumvachellianum industrialized seedling production method |
| CN106069712A (en) * | 2016-07-04 | 2016-11-09 | 广东海洋大学 | Suspend based on the Alga Sgrgassi Enerves seedling scale of amphigenetic Leizhou and aestivate breeding method |
| CN106857226A (en) * | 2017-03-03 | 2017-06-20 | 山东省海洋生物研究院 | A kind of method of Sargassum horneri in northern productivity artificial breeding |
| CN109105248A (en) * | 2018-08-29 | 2019-01-01 | 湖北工业大学 | A kind of indoor germination method of stonewort brood body |
| CN109197593A (en) * | 2018-10-29 | 2019-01-15 | 杭州园泰生物科技有限公司 | The fast breeding method of Wa Shi sargassum |
| CN110432138A (en) * | 2019-09-06 | 2019-11-12 | 广东海洋大学 | A method of accelerate reef film category seaweed frond mature |
| CN117136833A (en) * | 2023-08-18 | 2023-12-01 | 生态环境部华南环境科学研究所(生态环境部生态环境应急研究所) | Sargassum periwinkle germplasm preservation method |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103651118A (en) * | 2013-11-11 | 2014-03-26 | 青岛文创科技有限公司 | Method for sterilizing turbinaria conoides rhizoid explant |
| CN104115738A (en) * | 2014-07-29 | 2014-10-29 | 广东海洋大学 | Seedling method for inducing synchronous maturity of Leizhou gulfweed in vitro receptacles |
| CN105393909A (en) * | 2015-11-25 | 2016-03-16 | 上海海洋大学 | Sargassumvachellianum industrialized seedling production method |
| CN106069712A (en) * | 2016-07-04 | 2016-11-09 | 广东海洋大学 | Suspend based on the Alga Sgrgassi Enerves seedling scale of amphigenetic Leizhou and aestivate breeding method |
| CN106857226A (en) * | 2017-03-03 | 2017-06-20 | 山东省海洋生物研究院 | A kind of method of Sargassum horneri in northern productivity artificial breeding |
| CN109105248A (en) * | 2018-08-29 | 2019-01-01 | 湖北工业大学 | A kind of indoor germination method of stonewort brood body |
| CN109197593A (en) * | 2018-10-29 | 2019-01-15 | 杭州园泰生物科技有限公司 | The fast breeding method of Wa Shi sargassum |
| CN109197593B (en) * | 2018-10-29 | 2021-11-09 | 杭州园泰生物科技有限公司 | Rapid breeding method of gulfweed |
| CN110432138A (en) * | 2019-09-06 | 2019-11-12 | 广东海洋大学 | A method of accelerate reef film category seaweed frond mature |
| CN117136833A (en) * | 2023-08-18 | 2023-12-01 | 生态环境部华南环境科学研究所(生态环境部生态环境应急研究所) | Sargassum periwinkle germplasm preservation method |
| CN117136833B (en) * | 2023-08-18 | 2024-05-24 | 生态环境部华南环境科学研究所(生态环境部生态环境应急研究所) | Sargassum periwinkle germplasm preservation method |
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