CN100427581C - Process for solid culture of hair weeds cells - Google Patents
Process for solid culture of hair weeds cells Download PDFInfo
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- CN100427581C CN100427581C CNB2005101220599A CN200510122059A CN100427581C CN 100427581 C CN100427581 C CN 100427581C CN B2005101220599 A CNB2005101220599 A CN B2005101220599A CN 200510122059 A CN200510122059 A CN 200510122059A CN 100427581 C CN100427581 C CN 100427581C
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- hair weeds
- weeds cells
- culture
- dry
- cells
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Abstract
The present invention discloses a method for solid culture of nostoc flagelliform cells, which realizes the purpose that nostoc flagelliform can quickly grow relatively under the manual control condition or the natural condition. The present invention has the technical scheme that an activated nostoc flagelliform cell suspension is inoculated to the surface layer of a solid culture medium, a certain amount of culture liquid prepared in advance is sprayed on the solid culture medium, nostoc flagelliform cells are cultivated under the manual control condition or the natural condition, and are cultivated according to drying and wetting rhythm; the fully artificial culture or field amplification culture can be realized by controlling proper culture conditions. The technology of the present invention improves a traditional culture mode, and accelerates proliferation. The nostoc flagelliform cell cultures obtained by the present invention have strong drought resisting capability and can be preserved for a long time; in the process of growth, carbon dioxide in the air can be sucked, and nitrogen can be fixed by the nostoc flagelliform cell cultures which release oxygen to the atmosphere in order to perform the function of purifying the air; pesticides are not needed, and thus, toxic and harmful substances can not be generated. The amplification culture of the nostoc flagelliform cells is favorable for maintaining the ecological balance of arid regions and semi-arid regions and improving desertification soil, and thus, the present invention has high economic value and environmental protection value.
Description
Technical field
The present invention relates to a kind of cell culture technology, particularly a kind of hair weeds cells culture technique, but be specially the hair weeds cells process for solid culture of direct inoculation in the grains of sand or desert soil nature sandy soil top layer.
Background technology
(Nostoc flagelliforme) delivers vegetables, be a kind of prokaryotic organism, be under the jurisdiction of Prokaryota (Procaryotae), Cyanophyta (Cyanophyta), Cyanophyceae (Cyanophyceae), hormogon[ium order (Hormogonales), beads (Nostocaceae), Nostoc (Nostoc).It is nutritious, and has certain pharmaceutical use, has economic development value and potentiality, and because of the similar favor that is subjected to people for a long time to " making a good deal of money " of pronunciation.Deliver vegetables and mainly be distributed in the desert half-desert area of northwest and part provinces and regions, North China in China, deliver vegetables and to carry out photosynthesis, again can fixed nitrogen, be a kind of Lu Sheng blue-green algae that under the special ecological condition, lives, arid, high temperature, uv-radiation, severe environment such as barren are had very strong adaptability.Be fixed nitrogen resource main in the desert ecotope, have important ecological recovery effect.Since in the open air deliver vegetables the growth extremely slow, extensive in addition immoderate destructiveness is gathered its stock number is fallen sharply, and havoc grassland ecosystem.In order to protect hair-like seaweed resource and ecotope; for a long time; people attempt to adopt the method for artificial culture always; increase its biomass and stock number, satisfying the growing market requirement, and carried out a large amount of Study on Ecophysiological Characteristics of Narrow; for example; application number is: 02138828.8 and 03119101.0 Chinese patent application, carried out the work of hair weeds cells fluid suspension culture, and obtained some progress.But artificial culture never realizes fully.Present training is a material with the frond of delivering vegetables of field acquisition substantially, does not still have the report that utilizes hair weeds cells to carry out solid-state cultivation.
Summary of the invention
The present invention be directed to the research that the problems referred to above are carried out; its objective is that a kind of method is provided is simple; easy to operate; the protection of this endangered species and manually production help delivering vegetables; under the situation of making slow progress in the frond artificial culture of delivering vegetables at present; realize complete artificial culture, and can be under near the condition of nature or direct inoculation enlarge in the grains of sand or desert soil nature sandy soil top layer and produce, make the process for solid culture of its hair weeds cells that can grow relatively apace.
For achieving the above object, the technical scheme that the present invention solves the process for solid culture technical problem of described hair weeds cells is to have taked following technical measures:
1, nutrient solution: filling a prescription is NaNO
30.5-1.5g/L, MgSO
47H
2O 75mg/L, CaCl
22H
2O 30-40mg/L, NaSiO
3.9H
2O 55-65mg/L, K
2HPO
435-40mg/L, pH7.5-8.0.Nutrient solution nutritive salt uses chemical pure, is dissolved in the tap water to get final product.During preparation, avoid precipitation to generate as far as possible, little have precipitation also separate out can, nutrient solution need not to carry out high-temperature sterilization.
2, the preparation of hair weeds cells liquid: with the centrifugal or filtration of hair weeds cells liquid 4000rpm of liquid culture, abandoning supernatant is added fresh medium, make it to suspend again, perhaps dry hair weeds cells is soaked with nutrient solution, make it recover active, use as solid-state cultivation and plant.
3, grains of sand pre-treatment: 40 eye mesh screens sieve the grains of sand, wash, and 100 ℃ of oven dry are tiled in the tray, thick about 0.5-1.5cm.
4. hair weeds cells is cultivated: hair weeds cells liquid is pressed 400ml/m
2Inoculum size be inoculated in grains of sand top layer, press 1-1.5L/m
2Total consumption divide between photoperiod, spray nutrient solution or moisture 3 every days, cover transparent plastic film on the sand table and keep grains of sand top layer moistening.When unglazed the photograph, throw off film, make grains of sand superficial drying, do the wet rhythm and pace of moving things and cultivate.During open-air the cultivation, spray nutrient solution or water daytime when surface temperature is lower than 40 ℃, keep the soil layer top layer moistening, then keep dry state when night or surface temperature are higher than 40 ℃.
5, culture condition: temperature 24-26 ℃, illumination 60-180 μ molm
-2s
-1, light dark period 12h:12h; Or open-air natural condition are cultivated.
6, results: when the agglomerating or open-air cultivation of hair weeds cells growth arrives the harvest season on the sand table, go sand to gather.
7, drying: in dry below 45 ℃, air-dry or the oven dry.
8, preservation: drying at room temperature is preserved.
9, continue to cultivate: dry hair weeds cells is rehydration in new nutrient solution, can continue to cultivate.
Advantage of the present invention and positively effect are: owing to adopt the cell plate method, changed traditional frond training method, method is simple, and is easy to operate; Nutrient solution need not to carry out high-temperature sterilization, cultivates and to compare with the tradition frond of delivering vegetables, and the rate of propagation of delivering vegetables is faster, the protection and artificial production of this endangered species that helps delivering vegetables.Because having carried out doing the wet rhythm and pace of moving things cultivates, therefore, the present invention cultivates the hair weeds cells culture of acquisition, have and the wild similar stronger drought-resistant ability of delivering vegetables, and can be under drying conditions long-term preservation, can be directly used in production through enlarged culturing, be inoculated in the grains of sand or desert soil nature sandy soil top layer; Use this technology and cultivate hair weeds cells, but can absorb carbon dioxide in air and fixed nitrogen, in atmosphere, discharge oxygen, the effect that purifies air is arranged, and be different from modern agriculture production and will use agricultural chemicals and weedicide, need not any agricultural chemicals and weedicide in the production, can not produce any harmful toxic matter in its culturing process; This technological breakthrough traditional frond training method of delivering vegetables, its enlarged culturing helps keeping the improvement of the geographic eubiosis of arid and semi-arid and desertification soil, balance to grassland ecosystem has certain realistic meaning, and has the very high economic worth and the value of environmental protection.
Embodiment
Embodiment 1: the solid-state cultivation of hair weeds cells sand table, and its concrete grammar is:
A. prepare nutrient solution: NaNO
30.5g/L, MgSO
47H
2O 75mg/L, CaCl
22H
2O 30mg/L, NaSiO
3.9H
2O55mg/L, K
2HPO
435mg/L, pH7.5, mixing is standby.
B. prepare hair weeds cells liquid: hair weeds cells medium centrifugal or filtration, collect hair weeds cells.Add the ratio of 1L nutrient solution in 10g (weight in wet base) hair weeds cells, make it to suspend again, perhaps adopt dried hair weeds cells, the ratio that adds the 1L nutrient solution in 2.5g cell powder is soaked 24h, makes it to recover active, as solid-state cultivation with kind.
C. grains of sand pre-treatment: 40 eye mesh screens sieve the grains of sand, wash, and dry to constant weight for 100 ℃.Be tiled on glass, plastics, metal or the wooden tray thickness 0.5cm.
D. cell cultures: hair weeds cells liquid is pressed 400ml/m
2Inoculum size be inoculated in grains of sand top layer, press 1L/m
2Total consumption divide between photoperiod, spray nutrient solution or moisture 3 every days, cover transparent plastic film on the sand table and keep grains of sand top layer moistening.When unglazed the photograph, throw off film, make grains of sand superficial drying, do the wet rhythm and pace of moving things and cultivate.
E. culture condition: 24 ℃ of temperature, illumination 60 μ molm
-2s
-1, light dark period 12h:12h.
F. gather in the crops: when hair weeds cells is grown agglomerating or covered with the sand table top layer, go sand to gather.
G. dry: in dry below 45 ℃, air-dry or oven dry.
H. preservation: the hair weeds cells culture of collecting is preserved down in the drying at room temperature ventilated environment.
I. continue to cultivate:, can continue to cultivate with exsiccant hair weeds cells rehydration in new nutrient solution.
Embodiment 2:
A. prepare nutrient solution: NaNO
31.0g/L, MgSO
47H
2O 75mg/L, CaCl
22H
2O 35mg/L, NaSiO
39H
2O60mg/L, K
2HPO
437.5mg/L, pH7.25 (pH7.8), mixing is standby.
C. grains of sand pre-treatment: 40 eye mesh screens sieve the grains of sand, wash, and dry to constant weight for 100 ℃.Be tiled on glass, plastics, metal or the wooden tray thickness 1.0cm.
D. cell cultures: hair weeds cells liquid is pressed 600ml/m
2Inoculum size be inoculated in grains of sand top layer, press 1.2L/m
2Total consumption divide between photoperiod, spray nutrient solution or moisture 3 every days, cover transparent plastic film on the sand table and keep grains of sand top layer moistening.
E. culture condition: 25 ℃ of temperature, illumination 120 μ molm
-2s
-1, light dark period 12h:12h.
All the other are with embodiment 1.
Embodiment 3:
A. prepare nutrient solution: NaNO
31.5g/L, MgSO
47H
2O 75mg/L, CaCl
22H
2O 40mg/L, NaSiO
3.9H
2O65mg/L, K
2HPO
440mg/L, pH8.0, mixing is standby.
C. grains of sand pre-treatment: 40 eye mesh screens sieve the grains of sand, wash, and dry to constant weight for 100 ℃.Be tiled on glass, plastics, metal or the wooden tray thickness 1.5cm.
D. cell cultures: hair weeds cells liquid is pressed 800ml/m
2Inoculum size be inoculated in grains of sand top layer, press 1.5L/m
2Total consumption divide between photoperiod, spray nutrient solution or moisture 3 every days, cover transparent plastic film on the sand table and keep grains of sand top layer moistening.
E. culture condition: 26 ℃ of temperature, illumination 180 μ molm
-2s
-1, light dark period 12h:12h;
All the other are with embodiment 1.
Embodiment 4: hair weeds cells is open-air to be cultivated, and its concrete grammar is:
Hair weeds cells liquid is pressed 500ml/m
2Inoculum size be sprayed on the sandy soil top layer of desert-semi-desert grassland or hills bare area, cultivate under the natural condition.Sow every year by the end of April or at the beginning of 5 months, gathers by the end of November.Summer high temperature cultivation in season covers the sunshade net on the ground, to cover overgenerous illumination and to reduce surface temperature.Spray nutrient solution or water daytime when surface temperature is lower than 40 ℃, keep the soil layer top layer moistening, then keep dry state when night or surface temperature are higher than 40 ℃.All the other are with embodiment 1 (not comprising contents such as manually operated culture condition, grains of sand pre-treatment).
Embodiment 5: hair weeds cells is open-air to be cultivated, and its concrete grammar is:
Hair weeds cells liquid is pressed 750ml/m
2Inoculum size be sprayed on the sandy soil top layer of desert-semi-desert grassland or hills bare area, all the other are with embodiment 4.
Embodiment 6: hair weeds cells is open-air to be cultivated, and its concrete grammar is:
Hair weeds cells liquid is pressed 1000ml/m
2Inoculum size be sprayed on the sandy soil top layer of desert-semi-desert grassland or hills bare area, all the other are with embodiment 4.
Claims (2)
1. the process for solid culture of a hair weeds cells is characterized in that the hair weeds cells culture that fluid suspension culture obtains is inoculated in sandy soil matrix top layer, may further comprise the steps:
A. nutrient solution preparation: NaNO
30.5-1.5g/L, MgSO
47H
2O75mg/L, CaCl
22H
2O30-40mg/L, NaSiO
3.9H
2O 55-65mg/L, K
2HPO
435-40mg/L, pH7.5-8.0, mixing is standby;
B. hair weeds cells liquid prepares: hair weeds cells medium centrifugal or filtration, and the collection hair weeds cells adds fresh medium, makes it to suspend again, perhaps dry hair weeds cells is soaked with nutrient solution, makes it recover active, uses as solid-state cultivation and plants;
C. grains of sand pre-treatment: 40 eye mesh screens sieve the grains of sand, wash, and dry to constant weight for 100 ℃, are tiled on glass, plastics, metal or the wooden tray thickness 0.5-1.5cm;
D. hair weeds cells is cultivated: hair weeds cells liquid is inoculated in sandy soil matrix top layer; Press 1-1.5L/m photostage every day
2Total consumption divides and sprays nutrient solution or water 3 times, and sand table top layer coating film makes the matrix top layer keep moistening, unglazed according to the time then remove film, make the matrix superficial drying, do wet rhythm and pace of moving things cultivation; Culture temperature 24-26 ℃, illumination 60-180 μ molm
-2s
-1, light dark period 12h: 12h;
E. results: grow when hair weeds cells and to go sand to gather when agglomerating;
F. dry: in dry below 45 ℃, air-dry or oven dry;
G. preservation: the hair weeds cells drying at room temperature ventilated environment of collecting is preserved down.
2. the process for solid culture of a hair weeds cells is characterized in that the hair weeds cells culture that fluid suspension culture obtains is inoculated in sandy soil matrix top layer, may further comprise the steps:
A. nutrient solution preparation: NaNO
30.5-1.5g/L, MgSO
47H
2O75mg/L, CaCl
22H
2O30-40mg/L, NaSiO
3.9H
2O 55-65mg/L, K
2HPO
435-40mg/L, pH7.5-8.0, mixing is standby;
B. hair weeds cells liquid prepares: hair weeds cells medium centrifugal or filtration, and the collection hair weeds cells adds fresh medium, makes it to suspend again, perhaps dry hair weeds cells is soaked with nutrient solution, makes it recover active, uses as solid-state cultivation and plants;
C. hair weeds cells is cultivated: the sandy soil top layer that hair weeds cells liquid is sprayed on desert-semi-desert grassland or hills bare area, when being lower than 40 ℃, surface temperature sprays nutrient solution or water daytime, keep the soil layer top layer moistening, then keep dry state when night or surface temperature are higher than 40 ℃;
D. gather in the crops: sow every year by the end of April or at the beginning of 5 months, gathers by the end of November;
E. dry: in dry below 45 ℃, air-dry or oven dry;
F. preservation: the hair weeds cells drying at room temperature ventilated environment of collecting is preserved down.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005101220599A CN100427581C (en) | 2005-12-01 | 2005-12-01 | Process for solid culture of hair weeds cells |
| JP2006087903A JP4937620B2 (en) | 2005-12-01 | 2006-03-28 | Solid culture method of hair rape cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNB2005101220599A CN100427581C (en) | 2005-12-01 | 2005-12-01 | Process for solid culture of hair weeds cells |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1793314A CN1793314A (en) | 2006-06-28 |
| CN100427581C true CN100427581C (en) | 2008-10-22 |
Family
ID=36804964
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB2005101220599A Expired - Fee Related CN100427581C (en) | 2005-12-01 | 2005-12-01 | Process for solid culture of hair weeds cells |
Country Status (2)
| Country | Link |
|---|---|
| JP (1) | JP4937620B2 (en) |
| CN (1) | CN100427581C (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103976411B (en) * | 2014-04-30 | 2015-10-14 | 河南科技大学 | A kind of manufacture method of product of delivering vegetables |
| CN104593438A (en) * | 2015-01-19 | 2015-05-06 | 河南科技大学 | Extracting method of suspension-culture hair-like seaweed amino acid |
| CN107629984B (en) * | 2017-11-01 | 2020-10-30 | 山东省农业科学院试验基地服务中心 | Culture method of nostoc flagelliforme |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1096499C (en) * | 1999-10-22 | 2002-12-18 | 中国科学院水生生物研究所 | Method for culturing nostoc |
| CN1530439A (en) * | 2003-03-14 | 2004-09-22 | 天津科技大学 | Hair weeds bacterial culture and culturing product |
| CN1174090C (en) * | 2002-07-26 | 2004-11-03 | 中国科学院水生生物研究所 | Method for cultivating hair weeds cell |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0813715B2 (en) * | 1991-05-23 | 1996-02-14 | 株式会社荏原総合研究所 | Soil improvement method by soil algae |
| JP3573941B2 (en) * | 1998-02-13 | 2004-10-06 | マイクロアルジェコーポレーション株式会社 | Composition having hyaluronidase inhibitory activity and cosmetic containing the composition |
| JP2002068943A (en) * | 2000-08-30 | 2002-03-08 | Microalgae Corporation | TESTOSTERONE-5alpha-REDUCTASE INHIBITOR AND HAIR-GROWING AGENT CONTAINING THE SAME |
| JP2002069443A (en) * | 2000-08-30 | 2002-03-08 | Microalgae Corporation | Antioxidant and cosmetic containing the antioxidant |
| JP4339533B2 (en) * | 2001-08-17 | 2009-10-07 | マイクロアルジェコーポレーション株式会社 | Method for producing health food and health food obtained by the method |
| JP4532855B2 (en) * | 2003-06-26 | 2010-08-25 | 有限会社日本エコロノミックス | Method for preventing fine soil particles from flowing out |
-
2005
- 2005-12-01 CN CNB2005101220599A patent/CN100427581C/en not_active Expired - Fee Related
-
2006
- 2006-03-28 JP JP2006087903A patent/JP4937620B2/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1096499C (en) * | 1999-10-22 | 2002-12-18 | 中国科学院水生生物研究所 | Method for culturing nostoc |
| CN1174090C (en) * | 2002-07-26 | 2004-11-03 | 中国科学院水生生物研究所 | Method for cultivating hair weeds cell |
| CN1530439A (en) * | 2003-03-14 | 2004-09-22 | 天津科技大学 | Hair weeds bacterial culture and culturing product |
Non-Patent Citations (4)
| Title |
|---|
| culture of the terestrial cyanobacterium, nostocflagelliforme(cyanophyceae), under aquatic condition. Kunshan Gao and Changpeng Ye.J. Phycol,Vol.39 . 2003 |
| culture of the terestrial cyanobacterium, nostocflagelliforme(cyanophyceae), under aquatic condition. Kunshan Gao and Changpeng Ye.J. Phycol,Vol.39 . 2003 * |
| 发菜人工培养条件探索. 刘明志.湖南农业科学，第1期. 2005 |
| 发菜人工培养条件探索. 刘明志.湖南农业科学，第1期. 2005 * |
Also Published As
| Publication number | Publication date |
|---|---|
| JP2007151534A (en) | 2007-06-21 |
| CN1793314A (en) | 2006-06-28 |
| JP4937620B2 (en) | 2012-05-23 |
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