CN102041254A - Space breeding method of spirulina - Google Patents
Space breeding method of spirulina Download PDFInfo
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- CN102041254A CN102041254A CN2009101811455A CN200910181145A CN102041254A CN 102041254 A CN102041254 A CN 102041254A CN 2009101811455 A CN2009101811455 A CN 2009101811455A CN 200910181145 A CN200910181145 A CN 200910181145A CN 102041254 A CN102041254 A CN 102041254A
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- 240000002900 Arthrospira platensis Species 0.000 title claims abstract description 87
- 235000016425 Arthrospira platensis Nutrition 0.000 title claims abstract description 87
- 229940082787 spirulina Drugs 0.000 title claims abstract description 83
- 238000009395 breeding Methods 0.000 title abstract description 11
- 238000000034 method Methods 0.000 claims abstract description 15
- 238000000926 separation method Methods 0.000 claims abstract description 5
- 238000012216 screening Methods 0.000 claims abstract 3
- 241000195493 Cryptophyta Species 0.000 claims description 44
- 239000007788 liquid Substances 0.000 claims description 7
- 239000002609 medium Substances 0.000 claims description 5
- 239000007787 solid Substances 0.000 claims description 5
- 241000620196 Arthrospira maxima Species 0.000 claims description 4
- 238000012258 culturing Methods 0.000 claims description 4
- 239000001963 growth medium Substances 0.000 claims description 3
- 244000005700 microbiome Species 0.000 claims description 3
- 231100000350 mutagenesis Toxicity 0.000 claims description 3
- 230000000050 nutritive effect Effects 0.000 claims description 3
- 239000002775 capsule Substances 0.000 claims description 2
- 230000005486 microgravity Effects 0.000 claims description 2
- 238000002703 mutagenesis Methods 0.000 claims description 2
- 238000011084 recovery Methods 0.000 claims description 2
- 230000001488 breeding effect Effects 0.000 abstract description 7
- 235000016709 nutrition Nutrition 0.000 abstract description 2
- 230000035764 nutrition Effects 0.000 abstract description 2
- 230000035772 mutation Effects 0.000 abstract 3
- 238000004519 manufacturing process Methods 0.000 description 5
- 238000002360 preparation method Methods 0.000 description 5
- 230000000877 morphologic effect Effects 0.000 description 4
- 235000015097 nutrients Nutrition 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- 229930002875 chlorophyll Natural products 0.000 description 3
- 235000019804 chlorophyll Nutrition 0.000 description 3
- ATNHDLDRLWWWCB-AENOIHSZSA-M chlorophyll a Chemical compound C1([C@@H](C(=O)OC)C(=O)C2=C3C)=C2N2C3=CC(C(CC)=C3C)=[N+]4C3=CC3=C(C=C)C(C)=C5N3[Mg-2]42[N+]2=C1[C@@H](CCC(=O)OC\C=C(/C)CCC[C@H](C)CCC[C@H](C)CCCC(C)C)[C@H](C)C2=C5 ATNHDLDRLWWWCB-AENOIHSZSA-M 0.000 description 3
- 238000005516 engineering process Methods 0.000 description 3
- 238000004321 preservation Methods 0.000 description 3
- 102000004169 proteins and genes Human genes 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- VZCCETWTMQHEPK-UHFFFAOYSA-N gamma-Linolensaeure Natural products CCCCCC=CCC=CCC=CCCCCC(O)=O VZCCETWTMQHEPK-UHFFFAOYSA-N 0.000 description 2
- 235000020664 gamma-linolenic acid Nutrition 0.000 description 2
- 229960002733 gamolenic acid Drugs 0.000 description 2
- 239000004615 ingredient Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 235000021049 nutrient content Nutrition 0.000 description 2
- 238000005192 partition Methods 0.000 description 2
- 108010001515 Galectin 4 Proteins 0.000 description 1
- 102100039556 Galectin-4 Human genes 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000006835 compression Effects 0.000 description 1
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- 238000011156 evaluation Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000002360 explosive Substances 0.000 description 1
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- 238000013467 fragmentation Methods 0.000 description 1
- 238000006062 fragmentation reaction Methods 0.000 description 1
- VZCCETWTMQHEPK-QNEBEIHSSA-N gamma-linolenic acid Chemical compound CCCCC\C=C/C\C=C/C\C=C/CCCCC(O)=O VZCCETWTMQHEPK-QNEBEIHSSA-N 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
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- 230000004083 survival effect Effects 0.000 description 1
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- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The invention relates to a space mutation breeding method of spirulina and the spirulina obtained with the same. The method comprises the steps of space mutation, separation, screening, repeated breeding and the like, and the spirulina bred with the method has superior length, width, spiral distance, spiral width, spiral quantity and nutrition components in comparison with that of the mutation spirulina variety of a general spirulina variety.
Description
Technical field
The invention belongs to the biological induced-mutation field, be specifically related to spirulina space flight breeding method, and the spirulina that obtains of space flight breeding.
Background technology
Spirulina is subject to the influence of external environment factor and morphs.Find that in production process in the past under the culture condition of open type, some minus variants usually appear in spirulina, show as mainly that algal filament becomes straight, undesired spiral or algal filament diminishes, spiral is tight etc.In traditional physics and chemomorphosis, the mutagenesis odds is very little, and is that the probability that occurs of minus variant is much larger than plus variant generally speaking.Therefore, the present invention is intended to utilize the high and big characteristics of variation amplitude of space flight breeding aberration rate, the variation spirulina algae kind that cultivation makes new advances, and promptly wide, the spiral number of length, width, pitch, spiral shell, nutritive ingredient all are better than the variation algae kind of common algae kind.
Summary of the invention
The present invention has carried out the space breeding of spirulina algae kind, obtain having the variation spirulina algae kind of good character, the variation algae kind that obtains has been carried out biological preservation at China Committee for Culture Collection of Microorganisms common micro-organisms center, wherein spirulina plalensis spirulina (Spirulinaplatensis) deposit number after the variation is 3279, and preservation date is: on September 14th, 2009; Spirulina maxim after the variation (Spirulina maxima) deposit number is 3280, and preservation date is: on September 14th, 2009.Concrete breeding and selection are as follows:
1, from the laboratory of agricultural section spirulina production base, chooses spirulina maxim (Spirulinamaxima) and spirulina plalensis (Spirulina platensis) algae kind, the technical requirements of carrying the article preparation in strict accordance with recoverable spacecraft is sealed in spirulina maxim and spirulina plalensis algae kind in the centrifuge tube with solid Zarrouk substratum, carry China's the 20th st recoverable science and technology test satellite emission and go up space, after 18 days space flight, reclaim.
2, return agricultural section laboratory through the spirulina maxim and the spirulina plalensis algae kind of carrying, in sterilisable chamber, use liquid Zarrouk culture medium culturing.Take a sample and observe, the spirulina maxim after recovering to cultivate is separated respectively with spirulina plalensis and selects, select the variation algae kind that frond length, spiral number all are better than common algae kind, carry out individual plant and cultivate in microscopically.
3, the algae kind OD that cultivates when individual plant
560Reach 1.2 when above, adopt above partition method to carry out individual plant again and separate, repeat seed selection, form stable until each strain system.
Description of drawings
Fig. 1 is the microstructural photo of common blunt top filament, and magnification is respectively 16,40,160.
Fig. 2 is the microstructural photo of blunt top filament (PNK-2) behind the space flight, and magnification is respectively 16,40,160.
Fig. 3 is the microstructural photo of common very big filament, and magnification is respectively 16,40,160.
Fig. 4 is the microstructural photo of very big filament (MNK-7) behind the space flight, and magnification is respectively 16,40,160.
Embodiment
On August 27th, 2004, from the laboratory of agricultural section spirulina production base, extract spirulina maxim (Spirulina maxima) and spirulina plalensis (Spirulina platensis) algae kind, preserve with solid Zarrouk culture medium culturing.Before the algae kind is carried, press the technical requirements repetition test that recoverable spacecraft carries the article preparation, guarantee to carry safety.On September 27th, 2004, the technical requirements of carrying the article preparation in strict accordance with recoverable spacecraft is sealed in spirulina maxim and spirulina plalensis algae kind in the centrifuge tube with solid medium, carry China the 20th st recoverable science and technology test satellite (perigee 205km, higher apsis 350km, airborne period is 89.6min, 63 ° of orbital inclinations, the horizontal 10-4g of track microgravity; 10～30 ℃ of the interior temperature of recovery capsule) space in the emission, the space flight through 18 days was successfully reclaimed on October 15th, 2004.Described space treatment requires to be specially:
Safety inspection: carry nontoxic, no inflammable and explosive, the no nuclear radiation of article, no strong electromagnetic does not have harmful gas, leak of liquid.
Registration and check and accept: carry article by administrative authority's unitizing of the recoverable spacecraft side of manufacturing, numbering after, pack into and formulate the position.
Sample loads: pack into the preparation of satellite of sample should be finished at launching base, according to the aircraft launching procedure, requires load sample in the shortest time that emission allows.Requirement per sample, the surface detention time will be kept sample at non-growth conditions.Be used to cultivate transparent, the resistance to compression, corrosion-resistant of sample hose quality of algae kind.
Lift-launch is with the preparation of algae kind material: solid inclined-plane Zarrouk carries, and aseptic technique is inoculated in the 0.5ml centrifuge tube of the good substratum of prepared beforehand, and in the algae kind sample box of packing into, 0～5 ℃ of refrigerator is preserved.Pack in satellite specified location or carry in the general biological incubator of 40h before the emission.Satellite reclaims the inherent aseptic experiment chamber opening of back 30h general biological incubator, takes out sample box and cultivates centrifuge tube, sends lift-launch unit as early as possible back to.
On November 3rd, 2004, return agricultural section laboratory through the spirulina maxim and the spirulina plalensis algae kind of carrying.Take a sample and observe in microscopically: spirulina maxim and spirulina plalensis all have filament survival, and comparatively small amt mostly is the little algae section of 1～3 spiral.The algae kind that lift-launch is returned is cultivated with liquid nutrient medium in sterilisable chamber.
November 13, the algae liquid that can observe blunt top algae kind slightly is light green, and naked eyes can clearly be observed filament; Greatly the algae kind can obviously be seen the green algal filament of thick length.
November 23, low-power microscope is observed down: violent multipolarization variation has all taken place in spirulina maxim and spirulina plalensis through space treatment.
December 3, the spirulina maxim after recover cultivating is separated respectively with spirulina plalensis and selects.
The individual plant of spirulina separates employing microscopically single tube pipettor and selects separation method, promptly from spirulina medium, draw the algae liquid that has filament on a small quantity, in the Zarrouk nutrient solution, dilute, the algae liquid that takes a morsel again from diluent places on recessed of the list of microscopically, under low power lens, observe filament, choose individual plant target filament and change on another single recessed, be defined as behind single algal filament moving into and carry out the individual plant cultivation in the test tube of encoding, be equipped with 5ml Zarrouk nutrient solution in advance with 10 μ l pipettors.
Spirulina maxim is isolated 50 strains, and spirulina plalensis is isolated 30 strains.
When the algae kind being carried out the individual plant separation, mainly consider to select to have than big-difference with original algae kind on the form: the individuality that filament is long, sturdy.The individual increase, the probability that output improves are just big.The spirulina maxim spiral number of original algae kind is at 3～6, and spirulina plalensis spiral number is at 5～8.So select 10 of space spirulina plalensiss more than the spiral, 6 individualities more than the spiral of space spirulina maxim target in the material after recover cultivating as the seed selection of high yield and high quality algae kind.
The algae kind of coming out through seed selection is 12～18 of spirulina plalensis PNK-2 space strain spiral numbers wherein; Length 629.82～852.58 μ m, the common spirulina plalensis of comparison photograph increases by 166.52%; Pitch 50.93～59.00 μ m increase by 5.88%; Wide 17.46～20.44 μ m of spiral shell increase by 8.19%; Algal filament diameter 5.83～6.42 μ m increase by 9.48%.7～9 of spirulina maxim MNK-7 space strain spiral numbers; Length 613.65～814.82 μ m, the common spirulina maxim of comparison photograph increases by 77.18%; Pitch 98.60～108.13 μ m increase by 13.91%; Wide 29.29～34.49 μ m of spiral shell increase by 3.16%.The outdoor scale operation productivity of spirulina plalensis PNK-2 space strain reaches 10g/dm
2More than, comparison is according to kind of a growth by 21.7%.The common spirulina maxim of the productivity ratio contrast of spirulina maxim MNK-7 space strain increases by 27.12%.Spirulina plalensis PNK-2 space strain protein total content, Phycocyanins, C-, chlorophyll, β-Hu Luobusu etc. mainly include the common spirulina plalensis height that nutrition is all compared photograph, and wherein protein content is 69.57%.Protein total content, Phycocyanins, C-, chlorophyll, β-Hu Luobusu and the gamma-linolenic acid etc. of spirulina maxim MNK-7 space strain mainly include the common spirulina maxim height that nutrient inventory is all compared photograph, and wherein protein content is 71.56%.The gal4 amino acid evaluation of institute's seed selection space strain reaches 0.90, and security of products has reached national food hygienic standard.Present method has also obtained a collection of spirulina mutant strain with application potential, wherein has filament length to reach 3000～5000um, the spiral number reaches 50～80 spirulina maxim new lines.
The morphological specificity of the spirulina plalensis (PNK-2) after spirulina plalensis before the variation and the variation contrasts as depicted in figs. 1 and 2, and concrete data are referring to table 1.
Table 1 spirulina plalensis sets out the main morphological specificity of strain and PNK-2 relatively
μm
The morphological specificity of the spirulina maxim (MNK-7) after spirulina maxim before the variation and the variation contrasts as shown in Figure 3 and Figure 4, and concrete data are referring to table 2.
Table 2 spirulina maxim sets out the main morphological specificity of strain and MNK-7 relatively
μm
Main nutrient content before and after the variation is referring to table 3.
Table 3 main nutrient content relatively
| Kind | Protein | Chlorophyll | β-Hu Luobusu | Phycocyanins, C- | Gamma-linolenic acid |
| Common very big | 64.56％ | 0.92％ | ?0.13％ | 13.66％ | ?0.61％ |
| MNK-7 | 71.56％ | 1.18％ | ?0.18％ | 14.32％ | ?0.71％ |
| Common blunt top | 64.23％ | 0.93％ | ?0.15％ | 13.78％ | ?0.65％ |
| PNK-2 | 69.57％ | 1.01％ | ?0.16％ | 14.70％ | ?0.63％ |
As seen, the variation algae kind that through behind the space breeding, frond length, width, pitch, the spiral shell of spirulina maxim and spirulina plalensis algae kind is wide, spiral number, nutritive ingredient all are better than common algae kind.
Spirulina growth and breeding are mainly carried out in the fragmentation mode.OD when spirulina medium
560Value reaches 1.2 when above, and spirulina growth reaches logarithmic phase latter stage, and growth is in halted state substantially, and at this moment the form of frond is more stable.Therefore work as the algae kind OD that individual plant is cultivated
560Reach 1.2 when above, adopt above partition method to carry out individual plant again and separate, repeat seed selection, form stable until each strain system.
Find that in culturing process afterwards the very big strain of the space of 25 above spirals easily produces clustering phenomena, be difficult to disperse; And the algal filament ascent rate is too fast, is not suitable for present spirulina production technique.Therefore when the seed selection of high yield algae kind, do not consider the very big strain of space of 25 above spirals.
Claims (6)
1. the space flight method of a spirulina comprises space flight, separation screening, repetition seed selection step, it is characterized in that:
(1) space flight: choose colory spirulina algae kind and be sealed in the centrifuge tube, carry space on the satellites transmits, after 18 days space flight, reclaim with solid Zarrouk substratum;
(2) separation is selected: use liquid Zarrouk culture medium culturing through the spirulina algae kind after the mutagenesis in sterilisable chamber, take a sample and observe in microscopically, spirulina after recover cultivating is separated and screens, select the variation algae kind of frond length, in the Zarrouk nutritive medium, carry out individual plant and cultivate greater than common frond length;
(3) repeat seed selection: as the algae kind OD of individual plant cultivation
560Reaching 1.2 when above, carry out individual plant again according to above-mentioned screening criteria and separate, repeat seed selection, is form stable until each strain of algae kind.
2. the method for claim 1, it is characterized in that: the flying condition of described satellite is: perigee 205km, higher apsis 350km, airborne period are 89.6min, 63 ° of orbital inclinations, the horizontal 10-4g of track microgravity; Temperature is 10～30 ℃ in the recovery capsule.
3. method as claimed in claim 1 or 2 is characterized in that: described algae kind is spirulina plalensis algae kind or spirulina maxim algae kind.
4. method as claimed in claim 3 is characterized in that: screen the above spirulina plalensis individuality of 10 spirals, or 6 spirulina maxim individualities that spiral is above.
5. the spirulina plalensis spirulina (Spirulina platensis) that obtains by any described method of claim 1-4, depositary institution is China Committee for Culture Collection of Microorganisms common micro-organisms center, deposit number is 3279.
6. the spirulina maxim (Spirulina maxima) that obtains by any described method of claim 1-4, depositary institution is China Committee for Culture Collection of Microorganisms common micro-organisms center, deposit number is 3280.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910181145A CN102041254B (en) | 2009-10-12 | 2009-10-12 | Spirulina |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN200910181145A CN102041254B (en) | 2009-10-12 | 2009-10-12 | Spirulina |
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| CN102041254B CN102041254B (en) | 2012-09-05 |
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| CN200910181145A Expired - Fee Related CN102041254B (en) | 2009-10-12 | 2009-10-12 | Spirulina |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104498580A (en) * | 2014-12-05 | 2015-04-08 | 云南绿A生物产业园有限公司 | Method for measuring mechanical damage resistance and recovery ability of spirulina seeds and application |
| CN108265014A (en) * | 2017-12-28 | 2018-07-10 | 中国科学院南海海洋研究所 | One plant passes through high-quality seawater spirulina of space breeding acquisition and application thereof |
| CN108342344A (en) * | 2018-03-29 | 2018-07-31 | 浙江大学 | A method of increasing algal filament screw pitch and length improves growth of spirulina platensis carbon sequestration rate |
-
2009
- 2009-10-12 CN CN200910181145A patent/CN102041254B/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104498580A (en) * | 2014-12-05 | 2015-04-08 | 云南绿A生物产业园有限公司 | Method for measuring mechanical damage resistance and recovery ability of spirulina seeds and application |
| CN104498580B (en) * | 2014-12-05 | 2016-07-06 | 云南绿A生物产业园有限公司 | The assay method of a kind of spirulina algae kind resistant to mechanical damage and recovery capability and application |
| CN108265014A (en) * | 2017-12-28 | 2018-07-10 | 中国科学院南海海洋研究所 | One plant passes through high-quality seawater spirulina of space breeding acquisition and application thereof |
| CN108265014B (en) * | 2017-12-28 | 2019-06-25 | 中国科学院南海海洋研究所 | One plant of high-quality seawater spirulina and application thereof obtained by space breeding |
| WO2019129021A1 (en) * | 2017-12-28 | 2019-07-04 | 中国科学院南海海洋研究所 | High-quality seawater spirulina obtained by means of space-breeding and use thereof |
| US11021684B2 (en) | 2017-12-28 | 2021-06-01 | South China Sea Institute Of Oceanology, Chinese Academy Of Sciences | High-quality seawater Spirulina strain obtained by space-breeding and use thereof |
| CN108342344A (en) * | 2018-03-29 | 2018-07-31 | 浙江大学 | A method of increasing algal filament screw pitch and length improves growth of spirulina platensis carbon sequestration rate |
| CN108342344B (en) * | 2018-03-29 | 2020-10-20 | 浙江大学 | Method for increasing spiral pitch and length of algae filaments and improving growth carbon fixation rate of spirulina |
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