CN105684879A - Method for suspension culture of chondrus ocellatus - Google Patents
Method for suspension culture of chondrus ocellatus Download PDFInfo
- Publication number
- CN105684879A CN105684879A CN201610257663.0A CN201610257663A CN105684879A CN 105684879 A CN105684879 A CN 105684879A CN 201610257663 A CN201610257663 A CN 201610257663A CN 105684879 A CN105684879 A CN 105684879A
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- China
- Prior art keywords
- frond
- spore
- suspension culture
- chondrus ocellatus
- ocellatus holmes
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G33/00—Cultivation of seaweed or algae
-
- 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
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- Life Sciences & Earth Sciences (AREA)
- Marine Sciences & Fisheries (AREA)
- Environmental Sciences (AREA)
- Cultivation Of Seaweed (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a method for suspension culture of chondrus ocellatus. The method comprises the following steps: (1) sampling in a wild place; (2) treating frond; (3) releasing spores; (4) collecting the spores; (5) performing suspension culture of chondrus ocellatus seedlings. By adopting the method, the purpose of storing chondrus ocellatus over summer can be achieved, high stability and good controllability can be achieved, seed sources for large-scale planting popularization can be effectively obtained, relatively good vegetative reproduction can be achieved, the problem of germplasm degeneration can be alleviated due to generative propagation, and compared with conventional propagation, the method is easy in suspension culture harvesting, and the labor and the material can be saved.
Description
Technical field
The present invention relates to the suspension culture method of marine algae, specifically a kind of method of chondrus ocellatus Holmes suspension culture。
Background technology
At present, the chondrus ocellatus Holmes of China is also introduced into large-scale commercialization breeding production, still in semi and the state relying on collection resource, this yield that not only limit chondrus ocellatus Holmes and quality, and excessively gather the natural resources that also can threaten chondrus ocellatus Holmes, this problem to be solved and must rely on development artificial cultivation technique。Being faced with some problems in long-term cultivation: 1, along with the deterioration of sea environment, the kind matter of chondrus ocellatus Holmes is degenerated serious；2, spread needs to solve kind of a source problem 3；Chondrus ocellatus Holmes is faced with the problem crossing summer difficulty, if employing indoor cross the summer or freezer crosses the summer, huge labour cost, allows people hang back；4, the traditional cultivation pattern of marine nursery, nursery is unstable, poor controllability。
For promoting that chondrus ocellatus Holmes cultivation can be healthy, stable, lasting development, solve the practical problem in cultivation, it is necessary to solve two problems: one, set up the strain of inheritance stability；Two, realize semi-artificialization and arrive complete manual aquaculture model。
At present, Thallus Laminariae (Thallus Eckloniae), Thallus Laminariae and Thallus Porphyrae three are cultivated greatly Sargassum and all use the means adopting seedling from spore to carry out breeding of seed, technology is more ripe, but whole aquaculture model is suitable only for cultivating in sea area, recovery process is complicated, and running down along with sea environment, the quality of whole product receives very big impact。
Summary of the invention
A kind of method that it is an object of the invention to provide chondrus ocellatus Holmes suspension culture, with the problem solving to propose in above-mentioned background technology。
For achieving the above object, the present invention provides following technical scheme:
A kind of method of chondrus ocellatus Holmes suspension culture, comprises the following steps:
(1) field sampling
Chondrus ocellatus Holmes mainly dwells and is born in climax and brings on the reef of low tide band, during sampling, first select the frond that healthy color is bright-coloured, then frond is placed under light, determine the basic modes of reproduction of frond by observing the morphological characteristic of adult algae, then anhydrous low temperature takes back laboratory；
(2) frond processes
Take back the frond of laboratory, should process as early as possible, under inverted microscope, sterile razor blade is used to take the maturing part of its frond, then rinsing for several times with antiseptic sea water, surface epizoite is removed in aseptic cotton balls wiping, is finally placed in by frond in sterile petri dish and 1%VSE antiseptic sea water, at 10-15 DEG C, 10-100umol/m2Cultivate under s environment；
(3) spores release
After 50-70 minute, under inverted microscope, observe frond spore release situation, observe red circular bead in culture fluid and characterize diffusing of frond spore；
(4) spore is collected
Collect immediately after spore release, use the pipettor after wire drawing, under inverted microscope, draw spore, it is placed in aseptic culture plate and 1%VSE antiseptic sea water, repeatedly above-mentioned steps, until it reaches desired spore concentration at 10-15 DEG C, 10-200 μm of ol/m2Cultivate under s environment, change weekly one time of nutrition liquid；
(5) algae Seedling suspension culture
Treating that frond grows at least 1mm, scrape the algae Seedling of attachment with sterile scalpel gently, this process should be quick, it is to avoid frond pollutes, and uses the screen filtration of 125-250um/m order number, is gone to by algae Seedling in aseptic triangular flask, be 1%VSE antiseptic sea water in triangular flask；Triangular flask sealing ventilation is cultivated, and air-flow size ensures that frond can slowly roll, and is unlikely to again to be blown to by frond on bottle wall, changes weekly a culture fluid。
As the further scheme of the present invention: frond is placed under light in (1) by step, by observing the morphological characteristic of adult algae, determine that frond belongs to tetrasporaphite or egagametophyte, there is the vesicle of circle in described tetrasporaphite and frond, described egagametophyte and frond have the utricule grown nonparasitically upon another plant。
As the present invention further scheme: observing red circular bead in culture fluid in step (3) and characterize diffusing of frond spore, if occurring that frond breaks, a large amount of red circular beads are gushed out from frond, illustrate that frond has begun to release spore；If this phenomenon does not occur, often detect every other hour。
As the present invention further scheme: pipettor described in step (4) needs through high temperature wire drawing, make aperture reach single spore and pass through。
As the present invention further scheme: step (4) miospore culture vessel is the culture plate in 12 or 24 holes。
As the present invention further scheme: step (5) intermediate cam bottle uses air filter sealing ventilation to cultivate。
As the present invention further scheme: the condition of culture in step (5) is as follows: 20 DEG C, 1%VSE sea water is aseptic, 28ppt, 200-250 μm of ol/m2s。
Compared with prior art, the invention has the beneficial effects as follows: solve the preservation problem that chondrus ocellatus Holmes crosses in summer process；Stability is high, and controllability is strong；Effectively solving spread required kind source problem, relatively nourish and generate, syngenesis can alleviate kind of a matter degenerate problem；More traditional cultivation, suspension culture is gathered easily, can use manpower and material resources sparingly。
Detailed description of the invention
Below in conjunction with the embodiment of the present invention, the technical scheme in the embodiment of the present invention is clearly and completely described, it is clear that described embodiment is only a part of embodiment of the present invention, rather than whole embodiments。Based on the embodiment in the present invention, the every other embodiment that those of ordinary skill in the art obtain under not making creative work premise, broadly fall into the scope of protection of the invention。
Embodiment 1
(1) field sampling
First selecting the bright-coloured frond of healthy color (8-10g), be then placed under light by frond, it is determined that frond belongs to tetrasporaphite or egagametophyte, then anhydrous low temperature takes back laboratory；
(2) frond processes
Under inverted microscope, use sterile razor blade to take the maturing part of its frond, then rinse 10 times with antiseptic sea water, surface epizoite is removed in aseptic cotton balls wiping, finally frond is placed in sterile petri dish (1%VSE antiseptic sea water), at 15 DEG C, 100 μm of ol/m2Cultivate under s environment；
(3) spore release
After 50-70 minute, observing frond spore release situation under inverted microscope, if existing frond breaks, a large amount of red circular beads are gushed out from frond, illustrating that frond has begun to release spore, if there is not this phenomenon, often detecting every other hour；
(4) spore is collected
Collect immediately after spore release, use the pipettor after wire drawing (pin hole allows spore to pass through), spore is drawn under inverted microscope, it is placed in the culture plate (1%VSE antiseptic sea water) in aseptic 12 holes, repeatedly above-mentioned steps, until reaching desired spore concentration, 15 DEG C, 200 μm of ol/m2Cultivate under s environment, change weekly one time of nutrition liquid。
(5) algae Seedling suspension culture
Treat frond length extremely visible size (at least 1mm), the algae Seedling of attachment is scraped gently with sterile scalpel, this process should be quick, frond is avoided to pollute, use the screen filtration of specific 125um/m order number, algae Seedling is gone to (1%VSE antiseptic sea water) in aseptic triangular flask, triangular flask sealing ventilation (air filter) is cultivated, air-flow size ensures that frond can slowly roll, and is unlikely to again to be blown to by frond on bottle wall, changes weekly a culture fluid, condition of culture is as follows: 20 DEG C, 1%VSE sea water (aseptic, 28ppt), 250 μm of ol/m2s。
Embodiment 2
(1) field sampling
First selecting the bright-coloured frond of healthy color (8-10g), be then placed under light by frond, it is determined that frond belongs to tetrasporaphite or egagametophyte, then anhydrous low temperature takes back laboratory；
(2) frond processes
Under inverted microscope, use sterile razor blade to take the maturing part of its frond, then rinse 10 times with antiseptic sea water, surface epizoite is removed in aseptic cotton balls wiping, finally frond is placed in sterile petri dish (1%VSE antiseptic sea water), at 10 DEG C, 100 μm of ol/m2Cultivate under s environment；
(3) spore release
After 50-60 minute, observing frond spore release situation under inverted microscope, if existing frond breaks, a large amount of red circular beads are gushed out from frond, illustrating that frond has begun to release spore, if there is not this phenomenon, often detecting every other hour；
(4) spore is collected
Collect immediately after spore release, use the pipettor after wire drawing (pin hole allows spore to pass through), spore is drawn under inverted microscope, it is placed in the culture plate (1%VSE antiseptic sea water) in aseptic 12 holes, repeatedly above-mentioned steps, until reaching desired spore concentration, at 10 DEG C, 200umol/m2Cultivate under s environment, change weekly one time of nutrition liquid。
(5) algae Seedling suspension culture
Treat frond length extremely visible size (at least 1mm), the algae Seedling of attachment is scraped gently with sterile scalpel, this process should be quick, frond is avoided to pollute, use the screen filtration of specific 125um/m order number, algae Seedling is gone to (1%VSE antiseptic sea water) in aseptic triangular flask, triangular flask sealing ventilation (air filter) is cultivated, air-flow size ensures that frond can slowly roll, and is unlikely to again to be blown to by frond on bottle wall, changes weekly a culture fluid, condition of culture is as follows: 20 DEG C, 1%VSE sea water (aseptic, 28ppt), 250umol/m2s。
Embodiment 3
(1) field sampling
First selecting the bright-coloured frond of healthy color (8-10g), be then placed under light by frond, it is determined that frond belongs to tetrasporaphite or egagametophyte, then anhydrous low temperature takes back laboratory；
(2) frond processes
Under inverted microscope, use sterile razor blade to take the maturing part of its frond, then rinse 10 times with antiseptic sea water, surface epizoite is removed in aseptic cotton balls wiping, finally frond is placed in sterile petri dish (1%VSE antiseptic sea water), at 10 DEG C, 100umol/m2Cultivate under s environment；
(3) spore release
After 50-70 minute, observing frond spore release situation under inverted microscope, if existing frond breaks, a large amount of red circular beads are gushed out from frond, illustrating that frond has begun to release spore, if there is not this phenomenon, often detecting every other hour；
(4) spore is collected
Collect immediately after spore release, use the pipettor after wire drawing (pin hole allows spore to pass through), spore is drawn under inverted microscope, it is placed in the culture plate (1%VSE antiseptic sea water) in aseptic 12 holes, repeatedly above-mentioned steps, until reaching desired spore concentration, at 10 DEG C, 200umol/m2Cultivate under s environment, weekly more with changing one time of nutrition liquid。
(5) algae Seedling suspension culture
Treat frond length extremely visible size (at least 1mm), scrape the algae Seedling of attachment gently with sterile scalpel, this process should be quick, it is to avoid frond pollutes, use the screen filtration of specific 250um/m order number, algae Seedling is gone to (1%VSE antiseptic sea water) in aseptic triangular flask。Triangular flask sealing ventilation (air filter) is cultivated, and air-flow size ensures that frond can slowly roll, and is unlikely to again to be blown to by frond on bottle wall, changing weekly a culture fluid, condition of culture is as follows: 20 DEG C, and 1%VSE sea water is (aseptic, 28ppt), 250umol/m2s。
It is obvious to a person skilled in the art that the invention is not restricted to the details of above-mentioned one exemplary embodiment, and when without departing substantially from the spirit of the present invention or basic feature, it is possible to realize the present invention in other specific forms。Therefore, no matter from which point, embodiment all should be regarded as exemplary, and be nonrestrictive, the scope of the invention rather than described above limits, it is intended that all changes in the implication of the equivalency dropping on claim and scope included in the present invention。
In addition, it is to be understood that, although this specification is been described by according to embodiment, but not each embodiment only comprises an independent technical scheme, this narrating mode of description is only for clarity sake, description should be made as a whole by those skilled in the art, and the technical scheme in each embodiment through appropriately combined, can also form other embodiments that it will be appreciated by those skilled in the art that。
Claims (7)
1. the method for a chondrus ocellatus Holmes suspension culture, it is characterised in that comprise the following steps:
(1) field sampling
Chondrus ocellatus Holmes mainly dwells and is born in climax and brings on the reef of low tide band, during sampling, first select the frond that healthy color is bright-coloured, then frond is placed under light, determine the basic modes of reproduction of frond by observing the morphological characteristic of adult algae, then anhydrous low temperature takes back laboratory；
(2) frond processes
Take back the frond of laboratory, should process as early as possible, under inverted microscope, sterile razor blade is used to take the maturing part of its frond, then rinsing for several times with antiseptic sea water, surface epizoite is removed in aseptic cotton balls wiping, is finally placed in by frond in sterile petri dish and 1%VSE antiseptic sea water, at 10-15 DEG C, 10-100umol/m2Cultivate under s environment；
(3) spores release
After 50-70 minute, under inverted microscope, observe frond spore release situation, observe red circular bead in culture fluid and characterize diffusing of frond spore；
(4) spore is collected
Collect immediately after spore release, use the pipettor after wire drawing, under inverted microscope, draw spore, it is placed in aseptic culture plate and 1%VSE antiseptic sea water, repeatedly above-mentioned steps, until it reaches desired spore concentration at 10-15 DEG C, 10-200 μm of ol/m2Cultivate under s environment, change weekly one time of nutrition liquid；
(5) algae Seedling suspension culture
Treating that frond grows at least 1mm, scrape the algae Seedling of attachment with sterile scalpel gently, this process should be quick, it is to avoid frond pollutes, and uses the screen filtration of 125-250um/m order number, is gone to by algae Seedling in aseptic triangular flask, be 1%VSE antiseptic sea water in triangular flask；Triangular flask sealing ventilation is cultivated, and air-flow size ensures that frond can slowly roll, and is unlikely to again to be blown to by frond on bottle wall, changes weekly a culture fluid。
2. the method for chondrus ocellatus Holmes suspension culture according to claim 1, it is characterized in that, frond is placed under light in (1) by step, by observing the morphological characteristic of adult algae, determine that frond belongs to tetrasporaphite or egagametophyte, there is the vesicle of circle in described tetrasporaphite and frond, described egagametophyte and frond have the utricule grown nonparasitically upon another plant。
3. the method for chondrus ocellatus Holmes suspension culture according to claim 1, it is characterized in that, step (3) is observed red circular bead in culture fluid and characterizes diffusing of frond spore, if occurring that frond breaks, a large amount of red circular beads are gushed out from frond, illustrate that frond has begun to release spore；If this phenomenon does not occur, often detect every other hour。
4. the method for chondrus ocellatus Holmes suspension culture according to claim 1, it is characterised in that pipettor described in step (4) needs through high temperature wire drawing, makes aperture reach single spore and passes through。
5. the method for chondrus ocellatus Holmes suspension culture according to claim 1, it is characterised in that step (4) miospore culture vessel is the culture plate in 12 or 24 holes。
6. the method for chondrus ocellatus Holmes suspension culture according to claim 1, it is characterised in that step (5) intermediate cam bottle uses air filter sealing ventilation to cultivate。
7. the method for chondrus ocellatus Holmes suspension culture according to claim 1, it is characterised in that the condition of culture in step (5) is as follows: 20 DEG C, 1%VSE sea water is aseptic, 28ppt, 200-250 μm of ol/m2s。
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| CN201610257663.0A CN105684879A (en) | 2016-04-21 | 2016-04-21 | Method for suspension culture of chondrus ocellatus |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106106119A (en) * | 2016-07-11 | 2016-11-16 | 象山旭文海藻开发有限公司 | A kind of cultural method of Monostroma nitidum Wittr. spore collection massing |
| CN110122313A (en) * | 2019-06-13 | 2019-08-16 | 国家海洋环境监测中心 | Irish moss discoid body clone method for culturing seedlings |
| CN115443063A (en) * | 2020-04-01 | 2022-12-06 | 嘉吉公司 | Seaweed cultivation method and system |
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| US3879890A (en) * | 1974-02-06 | 1975-04-29 | Canadian Patents Dev | Algal polysaccharide production |
| CN1341349A (en) * | 2000-09-05 | 2002-03-27 | 上海众伟生化有限公司 | Artificial cultivation production method of chondrus crispus and production process of liquid carragheenan |
| CN103392584A (en) * | 2013-06-08 | 2013-11-20 | 中国海洋大学 | Seedling breeding method using asparagus spores |
| CN103858746A (en) * | 2014-03-27 | 2014-06-18 | 青岛崂好人海洋生物技术有限公司 | Seaweed cultivation method |
| CN104488690A (en) * | 2014-12-26 | 2015-04-08 | 中国科学院海洋研究所 | Gracilaria seaweed algal turf repairing method |
| CN104542237A (en) * | 2014-12-26 | 2015-04-29 | 中国科学院海洋研究所 | Seedling-raising method for gracilaria algae |
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2016
- 2016-04-21 CN CN201610257663.0A patent/CN105684879A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US3879890A (en) * | 1974-02-06 | 1975-04-29 | Canadian Patents Dev | Algal polysaccharide production |
| CN1341349A (en) * | 2000-09-05 | 2002-03-27 | 上海众伟生化有限公司 | Artificial cultivation production method of chondrus crispus and production process of liquid carragheenan |
| CN103392584A (en) * | 2013-06-08 | 2013-11-20 | 中国海洋大学 | Seedling breeding method using asparagus spores |
| CN103858746A (en) * | 2014-03-27 | 2014-06-18 | 青岛崂好人海洋生物技术有限公司 | Seaweed cultivation method |
| CN104488690A (en) * | 2014-12-26 | 2015-04-08 | 中国科学院海洋研究所 | Gracilaria seaweed algal turf repairing method |
| CN104542237A (en) * | 2014-12-26 | 2015-04-29 | 中国科学院海洋研究所 | Seedling-raising method for gracilaria algae |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106106119A (en) * | 2016-07-11 | 2016-11-16 | 象山旭文海藻开发有限公司 | A kind of cultural method of Monostroma nitidum Wittr. spore collection massing |
| CN110122313A (en) * | 2019-06-13 | 2019-08-16 | 国家海洋环境监测中心 | Irish moss discoid body clone method for culturing seedlings |
| CN115443063A (en) * | 2020-04-01 | 2022-12-06 | 嘉吉公司 | Seaweed cultivation method and system |
| CN115443063B (en) * | 2020-04-01 | 2024-05-31 | 嘉吉公司 | Seaweed cultivation method and system |
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Application publication date: 20160622 |