CN101326905A - Method for preserving Porphyra haitanensis young seedling at low-temperature - Google Patents
Method for preserving Porphyra haitanensis young seedling at low-temperature Download PDFInfo
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- CN101326905A CN101326905A CNA2008100715244A CN200810071524A CN101326905A CN 101326905 A CN101326905 A CN 101326905A CN A2008100715244 A CNA2008100715244 A CN A2008100715244A CN 200810071524 A CN200810071524 A CN 200810071524A CN 101326905 A CN101326905 A CN 101326905A
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- porphyra haitanensis
- haihanensis
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Abstract
The invention discloses a method for deep-freezing of Porphyra haihanensis seedlings, relating to Porphyra haihanensis, particularly relating to a method for deep-freezing of Porphyra haihanensis seedlings in the aquiculture field. A proper amount of refrigeration protectant is prepared; the refrigeration protectant is separately filled in a freezing storage tube; Porphyra haihanensis seedlings are collected; the collected Porphyra haihanensis seedlings are placed in the freezing storage tube filled with refrigeration protectant; and the freezing storage tube filled with refrigeration protectant is kept under cold storage. When resuscitation is required, the freezing storage tube filled with refrigeration protectant is placed in water for thawing and then taken out. The resuscitated Porphyra haihanensis seedlings kept under deep-freezing 180d can still have a survival rate above 95 percent after recovery cultivation. Because refrigeration protectant is used to preserve fresh Porphyra haihanensis seedlings, the processes of thallus separation, drying, cooling, etc. are required by conventional low-temperature preservation; thereby, the labor is saved to large extent, and the survival rate of Porphyra haihanensis thallus preserved under lower temperature is improved.
Description
Technical field
The present invention relates to a kind of Porphyra haitanensis, especially relate to a kind of deepfreeze store method of porphyra haitanensis seedling of aquaculture field.
Background technology
Porphyra haitanensis (Porphyra haitanensis) is the distinctive warm temperate zone property kind of China, is the important economic red algae that Fujian, Zhejiang and Guangdong Coastal are propagated artificially, and its output accounts for 80% of national laver gross yield.
The low temperature of Porphyra haitanensis algae kind is preserved, existing a large amount of research, the general employing processing of drying in the shade earlier to Porphyra haitanensis thallus now, be placed on-20～-30 ℃ of refrigerator and cooled and freeze the method for preservation, but when carrying out the research of Porphyra haitanensis genetic breeding, often need young shoot or seedling after mass disposal is tested end at every turn.Because seedling is smaller and fragile, the separation before the freezing and work of drying in the shade is cumbersome, workload is big.And along with the development of laver cell engineering seedling growing process, also often need to preserve a large amount of porphyra haitanensis seedlings, grow seedlings to be used for follow-up thallophytic enzymolysis.Therefore can adopt seedling band water directly to refrigerate, and reach and the thallus identical effect of refrigeration of drying in the shade, cause algologist's concern.At present, the fresh and alive band water-cooled of relevant thallus of porphyra is hidden, and does not meet report as yet.
Notification number provides a kind of seed selection Porphyra haitanensis breedings such as induced mutations, somatic cell clone and unisexual reproduction that utilize for the patent of invention of CN1565168, by the mutant of chemistry or the artificial mutagenesis acquisition of physics Porphyra haitanensis, cultivate into complete thallus with the single separation of mutant and with it with the biological tool enzyme.Height and their mutual ratio according to thallophytic main photosynthetic pigment and chromoprotein matter content, the free aminoacid content height, growth rate, indexs such as the thickness of frond and color and luster, filter out good mutant thallus, and with they once more enzymolysis obtain the cultivation of regenerating of single cells in-vitro.The single separation of immature thallus with obtaining is cultured to big thallus, utilizes unisexual reproduction to obtain the pure lines filamentous.The F1 of the pure lines filamentous of excellent strain is for thallus, its merit is behind genetic stability analysis and synthesis quality determination, pure lines free conchocelis to excellent strain carries out amplification cultivation, and transplant and in shell, make it to become shell conchocelis, subsequently, carry out carrying out that collect seedling in the land for growing field crops and culture experiment with shell conchocelis.Prove the stable excellent strain of proterties through field experiment, carry out long preservation and apply in the mode of free conchocelis.
Summary of the invention
The deepfreeze store method that the purpose of this invention is to provide a kind of porphyra haitanensis seedling.
The present invention includes following steps:
1) preparation refrigeration protectant;
2) will refrigerate protectant is sub-packed in the frozen pipe;
3) collect porphyra haitanensis seedling;
4) porphyra haitanensis seedling of collecting is placed be equipped with refrigeration protectant frozen pipe;
5) the protectant frozen pipe refrigeration of refrigeration will be housed.
It is the preparation of 25 ‰ sterilization seawater that the refrigeration protectant of being prepared preferably adopts salinity, percentage by volume, and glycerine is 10%, dimethyl sulfoxide (DMSO) (DMSO) 5%, glucose 15% surplusly is the sterilization seawater.Frozen pipe is preferably the frozen pipe of 50ml.Preferably adorn refrigeration protectant 30ml in the frozen pipe of each 50ml.Porphyra haitanensis seedling is preferably the fresh and alive porphyra haitanensis seedling of 1～3cm.Described refrigeration preferably will be equipped with the protectant frozen seal of tube of refrigeration and be placed-20～-25 ℃ of refrigerators to preserve.
The porphyra haitanensis seedling of collecting directly place be equipped with refrigeration protectant frozen pipe get final product, processing need not to dry in the shade.
When the needs recovery, the protectant frozen pipe of refrigeration will be housed place water-bath to thaw, can take out.Take out the porphyra haitanensis seedling in the frozen pipe, behind the sterilization seawer washing, can recover to cultivate.The temperature of water-bath is preferably 20 ℃, and the time of thawing in the water-bath can be 15min, best 2～4 times of the number of times of sterilization seawer washing.Porphyra haitanensis seedling after the recovery is cultivated through recovery, the fresh and alive porphyra haitanensis seedling of deepfreeze 180d, and survival rate still can reach more than 95%.
Because the present invention uses the refrigeration protectant that porphyra haitanensis seedling is carried out fresh and alive preservation; when therefore having avoided conventional low temperature to preserve; workload has been saved in the work such as processing that need to separate, to dry in the shade thallus greatly, has also improved the survival rate of Porphyra haitanensis thallus deepfreeze.
Embodiment
Suppose when the Porphyra haitanensis genetic breeding, by the diffusing of Porphyra haitanensis conchospore, sprout and obtained a large amount of porphyra haitanensis seedlings, can't in time carry out the individual plant cultivation to each strain porphyra haitanensis seedling, when investigating the economic characters of each strain Porphyra haitanensis, can adopt this method that the fresh and alive seedling of Porphyra haitanensis is carried out deepfreeze.Concrete steps are as follows:
1) be 25 ‰ sterilization seawater preparation refrigeration protectant with salinity, by volume percentage contains 10% glycerine, 5%DMSO, 15% glucose, 70% sterilization seawater;
The refrigeration protectant that 2) will prepare is got 30ml, is sub-packed in the frozen pipe of 50ml;
3) the fresh and alive seedling of the Porphyra haitanensis of 1～3cm is directly placed be equipped with refrigeration protectant frozen pipe;
4), place-20～-25 ℃ of refrigerators to preserve with behind the frozen seal of tube.
After storage a period of time, when needing to use these seedling, frozen pipe can be taken out the cultivation of recovering from refrigerator: frozen pipe is taken out, directly place 20 ℃ of water-baths to thaw from refrigerator; Behind the 15min, frozen pipe is taken out from water-bath,, can place blake bottle to carry out individual plant and cultivate through sterilization seawer washing 3 times.Holding time, with interior porphyra haitanensis seedling, survival rate can reach more than 95% at 180d.
Claims (6)
1. the deepfreeze store method of porphyra haitanensis seedling is characterized in that may further comprise the steps:
1) preparation refrigeration protectant;
2) will refrigerate protectant is sub-packed in the frozen pipe;
3) collect porphyra haitanensis seedling;
4) porphyra haitanensis seedling of collecting is placed be equipped with refrigeration protectant frozen pipe;
5) the protectant frozen pipe refrigeration of refrigeration will be housed.
2. the deepfreeze store method of porphyra haitanensis seedling as claimed in claim 1 is characterized in that described refrigeration protectant employing salinity is 25% sterilization seawater preparation, by volume percentage; glycerine is 10%; dimethyl sulfoxide (DMSO) 5%, glucose 15%, surplus is the sterilization seawater.
3. the deepfreeze store method of porphyra haitanensis seedling as claimed in claim 1 is characterized in that frozen pipe is the frozen pipe of 50ml.
4. the deepfreeze store method of porphyra haitanensis seedling as claimed in claim 3 is characterized in that dress refrigeration protectant 30ml in the frozen pipe of each 50ml.
5. the deepfreeze store method of porphyra haitanensis seedling as claimed in claim 1 is characterized in that porphyra haitanensis seedling is the fresh and alive porphyra haitanensis seedling of 1～3cm.
6. the deepfreeze store method of porphyra haitanensis seedling as claimed in claim 1 is characterized in that described refrigeration is to place-20～-25 ℃ of refrigerators to preserve with the protectant frozen seal of tube of refrigeration is housed.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100715244A CN101326905A (en) | 2008-08-01 | 2008-08-01 | Method for preserving Porphyra haitanensis young seedling at low-temperature |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008100715244A CN101326905A (en) | 2008-08-01 | 2008-08-01 | Method for preserving Porphyra haitanensis young seedling at low-temperature |
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| CN101326905A true CN101326905A (en) | 2008-12-24 |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102187855A (en) * | 2011-03-08 | 2011-09-21 | 中国农业大学 | Seedling refrigeration method for strawberry delayed cultivation |
| CN109757356A (en) * | 2019-03-25 | 2019-05-17 | 宁波大学 | A kind of porphyra haitanensis group thallus germplasm base construction method saved based on living body |
| CN110100718A (en) * | 2019-05-10 | 2019-08-09 | 山东省海洋生物研究院 | A kind of porphyra haitanensis cold net cultural method suitable for northern sea area |
| CN112075415A (en) * | 2020-09-25 | 2020-12-15 | 中国科学院海洋研究所 | Cryoprotectant and ultra-low temperature cryopreservation method for stichopus japonicus sperms |
-
2008
- 2008-08-01 CN CNA2008100715244A patent/CN101326905A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102187855A (en) * | 2011-03-08 | 2011-09-21 | 中国农业大学 | Seedling refrigeration method for strawberry delayed cultivation |
| CN109757356A (en) * | 2019-03-25 | 2019-05-17 | 宁波大学 | A kind of porphyra haitanensis group thallus germplasm base construction method saved based on living body |
| CN110100718A (en) * | 2019-05-10 | 2019-08-09 | 山东省海洋生物研究院 | A kind of porphyra haitanensis cold net cultural method suitable for northern sea area |
| CN110100718B (en) * | 2019-05-10 | 2021-06-25 | 山东省海洋生物研究院 | Porphyra haitanensis cold storage net culture method suitable for northern sea area |
| CN112075415A (en) * | 2020-09-25 | 2020-12-15 | 中国科学院海洋研究所 | Cryoprotectant and ultra-low temperature cryopreservation method for stichopus japonicus sperms |
| CN112075415B (en) * | 2020-09-25 | 2021-08-31 | 中国科学院海洋研究所 | Cryoprotectant and ultra-low temperature cryopreservation method for stichopus japonicus sperms |
| NL2028946A (en) * | 2020-09-25 | 2022-05-24 | Inst Oceanology Cas | Cryoprotective solution and ultra-low temperature cryopreservation method of stichopus japonicus sperm |
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Open date: 20081224 |