CN103865793A - Nostoc alga superficial layer culture system and culture method - Google Patents
Nostoc alga superficial layer culture system and culture method Download PDFInfo
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/02—Photobioreactors
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/34—Internal compartments or partitions
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
- C12M41/16—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature by recirculation of culture medium at controlled temperature
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- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/44—Means for regulation, monitoring, measurement or control, e.g. flow regulation of volume or liquid level
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention provides a nostoc alga superficial layer culture system and culture method. The system comprises a culture solution recovery tank (7), a multi-layered culture stand (1), a plurality of lighting devices (3), a recovered liquid filtering system (6) and at least one liquid dropping opening (12). The method comprises the following steps: placing a superficial layer culture disc on the multi-layered culture stand, wherein the alga species for inoculation is an alga species obtained by axenic cultivation with the diameter of 1-2mm; uniformly paving the alga species on the superficial layer culture disc during inoculation, wherein the inoculating density is controlled at 0.5g/cm<2> to 1g/cm<2>, and the inoculating thickness is 1cm-2cm; and spraying the culture solution to the culture and controlling the light and the temperature of the culture solution. The system does not need a special air pump to increase oxygen and carbon dioxide. The culture method is simple in step and cancels the processes such as ventilating, cleaning, changing the culture solution and the like. Over 60% of water can be saved, and over 50% of electricity can be saved. Pollution caused by impure algae can be effectively eliminated, and common nostoc is not easy to crush.
Description
Technical field
The invention belongs to biological technical field, specifically belong to a kind of Nostoc algae shallow-layer culture systems and cultural method.
Background technology
Nostoc formal name used at school " intend spherical nostoc " (
nostoc sphaeroids kuz), Nostoc is a kind of many cells colonies, and shape is take spherical as main, and there is a common colloid clothing sheath outside of its filament, is blackish green, and colony's diameter can reach 5 centimetres.The medical books such as " Mingyi Bielu ", Compendium of Material Medica, " property of medicine is examined ", " national herbal medicine compilation " think that Nostoc has improving eyesight benefit gas, control nyctalopia, antipyretic clearly every, the effects such as beauty treatment are scalded and are had concurrently in Ginseng Extract, treatment, because of its property of medicine gentleness, long-term edible effect with health care, beauty treatment.
Nostoc nutritive value is high, containing 18 seed amino acids, wherein eight seed amino acid rich contents of needed by human; In Nostoc dry-matter, total protein content is up to 56% left and right, and wherein 10 % are the Phycocyanins, C-that tool can anti-cancer and cancer-preventing function; The active polysaccharide content with raising body immunity reaches 12%; Many unsaturated thick resin acids are more than 10%; Chlorophyll 30.98 mg/g; Xitix 5.5mg/g, Determination of Vitamin C approaches fresh jujube, higher more than 5 times than hawthorn; VITMAIN B1, B2 are higher than general fungus algae; Containing 15 kinds, mineral substance, its calcium content, higher than general vegetables, is a kind of fabulous Natural Rich Calcium nutritive food.Because Nostoc has effect of dietotherapy, health care, nourishing, its nutrition proportion of composing meets human body needs very much, is " best protective foods " by world health organisation recommendations.
According to the Li Shanghao academician of aquatic institute of Chinese Academy of Sciences textual criticism, Nostoc is very harsh to the requirement of growing environment, and growth scope is very narrow, except the town of galloping along on horseback, Hefeng County, Hubei Province still has more than 10,000 mu, only has the distribution of minute quantity in Africa.Because its natural distributed is narrow, yield poorly, enjoyed for imperial family and high-level officials mainly as tribute, the food for the emperor in the past.In recent years along with a large amount of uses of agricultural chemicals, chemical fertilizer, add environmental pollution day by day serious, the growing environment of Nostoc is subject to very big destruction, the field of suitable Nostoc growth is fewer and feweri, output is more and more lower, five tons of its natural annual production less thaies, and resource has been on the verge of exhaustion.But thering is the special efficacies such as delicious food, dietotherapy, health care, nourishing due to Nostoc, market demand is increasing, causes the contradiction between supply and demand.
Relevant Nostoc artificial culture method studies have reported that: in Chinese patent literature CN1294190 (applying date: 1999.10.22) disclosed cultural method comprise prepared by nutrient solution formula and preparation, algae kind, aerated culture, sieving and grading, enhance mechanical strength, results and the link such as dry.In Chinese patent literature CN1528127 (applying date: 2003.10.10), disclosed method comprises the links such as prepared by nutrient solution preparation, algae kind, bio-reactor aerated culture, hardening treatment, screening results.Chinese patent literature CN101792716 (applying date: 2010.03.10) adopts semi-enclosed indoor pond to accomplish scale production, and discloses the structural principle in pond and the optimal conditions of cultivating 7 fundamentals of Nostoc.Chinese patent literature CN201424471 (applying date: 2009.06.09) discloses a kind of air lift unit and Nostoc artificial culture device of cultivating for Nostoc, the high dispersing that produces in culturing process is stingy, and bag is spread in nutrient solution, makes Nostoc form regular rise and fall campaign in process of growth.
No matter the culture vessel that above patent documentation uses in culturing process is glass jar, bio-reactor, pond or raceway pond, all need Nostoc to mix with nutrient solution, in whole culturing process, must ventilate or impeller stirs Nostoc is suspended in nutrient solution by air pump, and provide CO for Nostoc
2and O
2, in the culture cycle of nearly 90 days, needed clean frond and change nutrient solution one time every 7～10 days; And changing in nutrient solution process, it is a very common problem that Nostoc spheroid breaks, general percentage of damage is between 5%-10%, broken serious more than 10%, especially large at air-flow or stir violent in the situation that and even can make Nostoc spheroid all disintegrate.In addition, general cultural method water power expends greatly, cost is high.
Summary of the invention
For water power in Nostoc culturing process in prior art expend greatly, easily broken, be subject to assorted algae pollution, the high problem of human cost, the application aims to provide a kind of Nostoc algae shallow-layer culture systems and cultural method.This system is measured body design according to the fostering requirement of Nostoc, simple in structure, can directly carry out temperature control to nutrient solution, and recycle, and does not need special air pump to carry out oxygenation and carbonic acid gas, is suitable for Nostoc algae culture.This cultural method can be economized on water more than 60%, and brownout more than 50%, can effectively be removed assorted algae pollution, and Nostoc is difficult for broken in culturing process.
For achieving the above object, technical scheme of the present invention is:
Nostoc algae shallow-layer culture systems, comprise nutrient solution pond for recovering, also comprise by multiple passive vertical support bars and multiple Multilayer culture shelf, multiple illumination apparatus, recovery liquid filtering system, nutrient solution delivery controller, nutrient solution transport pipe, at least one liquid droping port forming for placing the horizontal support bar of shallow-layer culture plate; Described illumination apparatus is arranged on the below of any one horizontal support bar of Multilayer culture shelf, and described Multilayer culture shelf bottom is provided with nutrient solution recovering tray, and described nutrient solution recovering tray is communicated with nutrient solution pond for recovering by recovery channel, recovery liquid filtering system; Described nutrient solution pond for recovering is provided with nutrient solution transferpump and temperature control device in being communicated with; Described nutrient solution transferpump is communicated with liquid droping port by nutrient solution transport pipe, and described liquid droping port is positioned at the top of Multilayer culture shelf top layer; Described shallow-layer culture plate bottom is provided with multiple holes, in described hole, is equipped with sealing plug, is provided with a Height Adjustable liquid level control minim pipette in the middle of described sealing plug.
On described nutrient solution transport pipe, be preferably provided with nutrient solution delivery controller.
The aperture of described shallow-layer culture plate bottom is preferably 0.8-1.2 cm.
The caliber of described liquid level control minim pipette is preferably 1mm-2 mm, and pipe range is preferably 4cm-8cm, and the caliber of described liquid level control minim pipette is less than the diameter of cultivated algae kind.
The interlamellar spacing of described Multilayer culture shelf is preferably 30 cm-40cm, and interlamellar spacing is adjustable.
The distance of described liquid droping port and Multilayer culture shelf top layer is preferably 40cm-50cm.
Nostoc algae shallow-layer cultural method, uses described culture systems, and concrete steps are:
(1) take glasshouse or plastic greenhouse greenhouse as cultivating place, described culture systems is placed in and is cultivated in place, shallow-layer culture plate is placed on Multilayer culture shelf, and the every interlayer of Multilayer culture shelf is divided into 30 cm～40cm, and the distance that liquid droping port and Multilayer culture shelf top layer are set is 40cm-50cm; Described shallow-layer culture plate bottom is provided with multiple holes, and shallow-layer culture plate aperture is 0.8-1.2 cm, in described hole, is equipped with sealing plug, is provided with a Height Adjustable liquid level control minim pipette in the middle of described sealing plug;
(2) inoculation is the algae kind that the diameter that obtains by sterile culture is 1mm～2mm by algae kind, when inoculation, algae kind is evenly laid in shallow-layer culture plate, and the density of inoculation is controlled at 0.5 g/cm
2～1g/cm
2, inoculation thickness is 1 cm-2cm;
(3) select diameter to be less than the liquid level control minim pipette of Nostoc particle diameter according to the diameter of culture, regulate liquid level control minim pipette height on culture plate, make the nutrient solution liquid level drenching into culture plate just flood culture, unnecessary nutrient solution flows into lower floor's culture plate by liquid level control minim pipette, automatically dissolves in fresh air when nutrient solution splashes into culture plate; Meanwhile, controlled light remains between 1000lx～20000lx, controls culture-liquid temp and remains between 20 ℃～30 ℃; Described nutrient solution composition and concentration are:
MgSO
4. 7H
2O 0.05-0. 15g/L
K
2HPO
4. 3H
2O 0.005-0.08g/L
CaCl
2. 2H
2O 0.004-0.06g/L
EDTA-Na
2 0.001-0.004g/ L
Ferric ammonium citrate 0.001-0.005 g/ L
H
3BO
4 0.002-0.008mg/L
Na
2MoO
4.2H
2O 0.001-0.030mg/L
ZnSO
4.7H
2O 0.001-0.02mg/L
CuSO
4.5H
2O 0.001-0.005mg/L
MnCl
2.4H
2O 0.020-0.060mg/L。
Described Nostoc algae is Nostoc or Nostoc commune.
The present invention will be further explained below:
Nostoc is cultivated and is comprised following step: nutrient solution component and preparation, and light and temperature condition control, algal species cultivation, inoculates, and cultivates, and nutrient solution reclaims and processes, results and storage.
A. take glasshouse or plastic greenhouse greenhouse as cultivating place, make full use of natural light temperature resource; Simultaneously top, greenhouse arrange can auto-switch sunshade net, greenhouse in the certain artificial lighting equipment of interpolation, make indoor illumination remain on 1000～20000lx; Directly control culture-liquid temp by temperature controlling instrumentss such as geothermal source air-conditionings, and by dripping the speed of nutrient solution, culture temperature is remained on to 20～30 ℃, effectively reduce energy consumption.
B. culturing rack framework is made by aluminium alloy, and every interlayer is divided into 30～40cm, and according to the illumination particularity of Various Seasonal, spacing distance between regulating course, makes intensity of illumination remain on optimum range.Nutrient solution catch tray is put in culturing rack bottom, and is connected with nutrient solution recovery channel, to reclaim filtration nutrient solution.Shallow-layer culture plate by nontoxic transparent material make, diameter glass tube 1-4mm, nostoc sphaeroides is put into shallow-layer culture plate after hierarchal arrangement on culturing rack, regulate as required nutrient solution flow velocity, make culture keep optimal temperature.
C. the basic composition of nutrient solution and concentration are the same.Nutrient solution formula need adjust accordingly at the different growth phases of Nostoc, and the nutrient solution of recovery need be supplemented according to the loss of nutrition in the time recycling.
D. inoculate and should be the good algae kind that the diameter that obtains by sterile culture is 1～2mm by algae kind, when inoculation, algae kind is evenly laid in shallow-layer culture plate, the density of inoculation is controlled at 0.2～1g/cm
2, carry out certain adjustment with season and illumination power.
E. the water dropper of being installed by two-position controller control culturing rack top continues to drip nutrient solution to culture, and nutrient solution flow rate regulation is to keep culture-liquid temp in culture plate to be advisable at 20～25 ℃.Nutrient solution is dripped to lower floor by upper strata, finally flow to the nutrient solution catch tray of the lowest layer, then is recovered to nutrient solution recovery system.Select diameter to be slightly less than the Glass tubing of Nostoc particle diameter according to the diameter of culture, regulate culture plate upper glass pipe height, make the nutrient solution liquid level drenching into culture plate just flood culture, unnecessary nutrient solution flows into lower floor's culture plate by Glass tubing, when nutrient solution splashes into culture plate, can automatically dissolve in fresh air.
F. the nutrient solution reclaiming need filter out impurity and flow into pond for recovering, and then by temperature control system, culture-liquid temp is controlled to 20～25 ℃ in pond for recovering, then re-uses by sprinkling equipment.
G. when the Nostoc stand density of cultivation is excessive, to divide dish.
Shallow-layer cultural method and the culture systems of described Nostoc, be also applicable to the scale operation of other algae of Nostoc such as Nostoc commune.
Nostoc ecological simulation culture systems provided by the present invention, in the time carrying out the production of mass-producing cultivation Nostoc, only need be put into nostoc sphaeroides shallow-layer culture plate, is placed on culturing rack and cultivates.
Many researchists drop into a large amount of manpower and materials the artificial culture method of Nostoc are studied in recent years, at present, all artificial culture methods about Nostoc all comprise prepared by nutrient solution preparation, algae kind, aerated culture, hardening treatment, screening results and the link such as dry, and culture vessel is glass jar or raceway pond.These methods all exist production link many, and water loss is large, and energy consumption is high, and spheroid easily breaks and the problem such as production cost height.Therefore we have designed the raw shallow-layer culture systems of brand-new Nostoc and method, can effectively address these problems.
No matter the culture vessel using is glass jar, bio-reactor, pond or raceway pond, all need Nostoc to mix with nutrient solution, in whole culturing process, must Nostoc be suspended in nutrient solution by ventilation or impeller, in the culture cycle of nearly 90 days, needed to change a nutrient solution every 7～10 days.In new culture systems, we simplify step, removed ventilation, have cleaned, change the processes such as nutrient solution, can save a large amount of manpowers, effectively increase work efficiency, and reduce production costs.
In new culture systems, we rest on Nostoc in culture plate and do not need Nostoc to carry out suspension culture, thereby can economize on water more than 70%; Temperature control neither be undertaken by controlling whole culture space, just can reach and only need carry out temperature control to nutrient solution the temperature condition that makes culture remain on suitable growth, thereby can energy efficient more than 50%.
In new culture systems, can effectively suppress assorted algae pollution, can reduce compared with traditional method and pollute more than 90%.
In culturing process, to break be an insoluble problem to Nostoc spheroid always.This is mainly owing to needing to pass into large quantity of air in culturing process, Nostoc to be suspended in nutrient solution, and the violent surging force of air-flow and make Nostoc spheroid subject to damage with the friction of culture vessel, broken serious, general percentage of damage is between 5%-10%, broken serious more than 10%, in the time changing nutrient solution, even can all break.And in the process that use present method is cultivated, Nostoc spheroid is seldom damaged, damaged lower than 3%.
Compared with prior art, advantage of the present invention is:
1, the production system of Nostoc is cultivated in mass-producing provided by the present invention, does not need whole culture space to be incubated, and only need carry out temperature control to nutrient solution just can reach the required culture temperature requirement of Nostoc, thereby energy consumption is little; Do not rely on pneumatic pump suspension frond, dripped in the journey of lower floor's culture plate because nutrient solution and air fully contact and dissolved in large quantity of air the Nostoc required CO that grows is enough provided at nutrient solution
2and O
2; Because the aperture of the posticum in culture plate design is less than the diameter of Nostoc, thereby in the time that nutrient solution goes out to underflow, can make various other pollutents that are less than Nostoc along with current flow away, thereby can play the effect that reduces assorted algae pollution; Be that shallow-layer is cultivated due to what adopt, culture density is large, and required nutrient solution is few, thereby can save more than 60% nutrient solution than common cultural method.It is simple that Nostoc shallow-layer culture systems is produced the required technology and equipment of Nostoc, invests littlely, and production cost is low and can annually produce continuously.
2, cultural method of the present invention, is to leave standstill culture method because of what adopt, and the surging force that spheroid is subject to is little, and spheroid is difficult for breaking.Light, the warm subject range of Nostoc are wider, and temperature is at 4-35 ℃, and illumination all can be grown at 600-12000lx, and photosynthetic efficiency is high, this cultural method algae kind fast growth, and in culturing process, fresh weight Average weight increasing a day can reach more than 6%.The assorted algae pollution rate of Nostoc is low, good stress resistance.
3, cultural method of the present invention, owing to only culture-liquid temp being controlled, can effectively reduce energy consumption, and save energy reaches more than 50%; In culturing process, can economize on water more than 70%; Nutrient solution composition and potential of hydrogen, close to natural water, can be recycled, and directly discharge does not pollute the environment yet, and the method is simplified step, has removed ventilation, has cleaned, changes the processes such as nutrient solution, effectively increases work efficiency, and reduces production costs.
Accompanying drawing explanation
Fig. 1 culture systems structural representation of the present invention;
Fig. 2 shallow-layer culture plate of the present invention structure for amplifying schematic diagram;
Wherein, the 1st, Multilayer culture shelf, the 2nd, shallow-layer culture plate, the 3rd, illumination apparatus, the 4th, nutrient solution recovering tray, the 5th, recovery channel, the 6th, reclaim liquid filtering system, the 7th, nutrient solution pond for recovering, the 8th, nutrient solution transferpump, the 9th, temperature control device, the 10th, nutrient solution delivery controller, the 11st, nutrient solution transport pipe, the 12nd, liquid droping port, the 13rd, vertical support bar, the 14th, horizontal support bar, the 15th, sealing plug, the 16th, liquid level control minim pipette, the 17th, algae kind, the 18th, nutrient solution.
Embodiment
Below in conjunction with embodiment, the present invention is described further.
Nostoc algae shallow-layer culture systems, comprise nutrient solution pond for recovering 7, also comprise by multiple passive vertical support bars 13 and multiple Multilayer culture shelf 1, multiple illumination apparatus 3, recovery liquid filtering system 6, nutrient solution delivery controller 10, nutrient solution transport pipe 11, at least one liquid droping port 12 forming for placing the horizontal support bar 14 of shallow-layer culture plate 2; Described illumination apparatus 3 is arranged on the below of any one horizontal support bar 14 of Multilayer culture shelf 1, described Multilayer culture shelf 1 bottom is provided with nutrient solution recovering tray 4, and described nutrient solution recovering tray 4 is communicated with nutrient solution pond for recovering 7 by recovery channel 5, recovery liquid filtering system 6; In being communicated with, described nutrient solution pond for recovering 7 is provided with nutrient solution transferpump 8 and temperature control device 9; Described nutrient solution transferpump 8 is communicated with liquid droping port 12 by nutrient solution transport pipe 11, and described liquid droping port 12 is positioned at the top of Multilayer culture shelf 1 top layer; Described shallow-layer culture plate 2 bottoms are provided with multiple holes, are equipped with sealing plug 15 in described hole, in the middle of described sealing plug 15, are provided with a Height Adjustable liquid level control minim pipette 16.
Described nutrient solution transport pipe 11 is provided with nutrient solution delivery controller 10.The aperture of described shallow-layer culture plate 2 bottoms is 0.8-1.2 cm.The caliber of described liquid level control minim pipette 16 is 1mm-2 mm, pipe range 4cm-8cm, and the caliber of described liquid level control minim pipette 16 is less than the diameter of cultivated algae kind.
The interlamellar spacing of described Multilayer culture shelf 1 is 30 cm-40cm, and interlamellar spacing is adjustable.Described liquid droping port 12 is 40cm-50cm with the distance of Multilayer culture shelf 1 top layer.
Take Nostoc as example, Nostoc ecological simulation cultural method, cultivates by culture systems described in embodiment 1, specifically comprises:
Cultivate place and equipment: take glasshouse or plastic greenhouse greenhouse as cultivating place, described culture systems is placed in and is cultivated in place, shallow-layer culture plate is placed on Multilayer culture shelf, the every interlayer of Multilayer culture shelf is divided into 30 cm～40cm, shallow-layer culture plate aperture 1-4mm, the distance that liquid droping port and Multilayer culture shelf top layer are set is 40cm-50cm;
Light source: the Nostoc required light intensity of growing is 1000-8000lx, can, without source of artificial light, in the time that light intensity is too high, need reduce light intensity by sunshade net in the time that natural light light intensity meets this condition.Ordinary Light Sources Have is water-proof daylight lamp pipe or LED lamp, and these lamps are arranged under the horizontal frame of culturing rack, can obtain the irradiation of 1000-8000lx light intensity to maintain frond.
Temperature: the growth temperature range of Nostoc is 10-35 ℃, optimum temps is 20-25 ℃ of left and right.In the time that natural temperature is in this temperature range, only need open greenhouse side form and teleblem carries out ventilation; When temperature is too low or too high, need be by regulating culture-liquid temp and flow velocity to make culture keep suitable temperature.
Nutrient solution composition and concentration are:
MgSO
4. 7H
2O 0.05-0. 15g/L
K
2HPO
4. 3H
2O 0.005-0.08g/L
CaCl
2. 2H
2O 0.004-0.06g/L
EDTA-Na
2 0.001-0.004g/ L
Ferric ammonium citrate 0.001-0.005 g/ L
H
3BO
4 0.002-0.008mg/L
Na
2MoO
4.2H
2O 0.001-0.030mg/L
ZnSO
4.7H
2O 0.001-0.02mg/L
CuSO
4.5H
2O 0.001-0.005mg/L
MnCl
2.4H
2O 0.020-0.060mg/L。
Prepared by algae kind: nostoc sphaeroides divides two kinds of firsts and seconds.When wherein prepared by First stage algae seeds, first prepare nutrient solution with distilled water, then divide and install to 50-l000ml Erlenmeyer flask or reagent bottle, through 121 ℃ of sterilizings of high pressure 20 minutes, cooling for subsequent use after sealing; Then select vigorous, the shapeliness of growth, good springiness and free of contamination Nostoc spheroid, in laboratory, under strict aseptic condition, carry out disinfection, be inoculated into the substratum preparing after homogenate, pass into filtrated air and carry out aerated culture; When cultivation, temperature is controlled at 20-25 ℃, and illumination is controlled at 3000-5000lx, and adds appropriate ruddiness.In the time that frond is cultivated diameter 0.5-1mm, be transferred in 10-20L carboy, carry out enlarged culturing, culture condition is the same; In the time that frond diameter grows to 2mm left and right, check good, the pollution-free secondary algae kind production kind that can be used as of growth conditions).
Inoculation and cultivation: the fine quality production kind of preparing by above method is put into shallow-layer culture plate, and the density of inoculation is controlled at 0.3 g/cm
2～1g/cm
2, inoculation thickness is 0.5-2cm, then the layering of shallow-layer culture plate is placed on culturing rack, adds nutrient solution by certain flow rate, and culture-liquid temp is controlled to 20-25 ℃; Cross and should or supplement illumination by source of artificial light by sunshade when strong or not enough and make it meet culture condition in illumination.Select diameter to be less than the liquid level control minim pipette of Nostoc particle diameter according to the diameter of culture, regulate liquid level control minim pipette height on culture plate, make the nutrient solution liquid level drenching into culture plate just flood culture, unnecessary nutrient solution flows into lower floor's culture plate by liquid level control minim pipette, automatically dissolves in fresh air when nutrient solution splashes into culture plate; The nutrient solution dripping is successively leaked down to lower floor by upper strata, finally fall into undermost nutrient solution catch tray, export and flow into collection tube by collector tray bottom again, equipment enters nutrient solution collecting tank after filtering after filtration, by temperature controlling instruments, culture-liquid temp is adjusted to 20-25 ℃ of winter temperature more slightly high, summer temp is slightly low, then re-uses by drip irrigation system.In collecting tank, the concentration of nutrient solution composition must be monitored, and loss part must be supplied in time.In culturing process, in the time that Nostoc increment is double, assign in two culture plates and to continue to cultivate, by that analogy, until Nostoc grows into 6-8mm, cross after screen sizing direct marketing or dry in the shade after sell.
This cultural method algae kind fast growth, in culturing process, fresh weight Average weight increasing a day can reach more than 6%.
Product detects: by being observed visually the Nostoc spheroid situation of not breaking, effectively suppressed assorted algae pollution, reduced and pollute more than 70% compared with traditional method.
Claims (8)
1. Nostoc algae shallow-layer culture systems, comprise nutrient solution pond for recovering (7), it is characterized in that, also comprise by multiple passive vertical support bars (13) and multiple Multilayer culture shelf (1), multiple illumination apparatus (3), recovery liquid filtering system (6), nutrient solution delivery controller (10), nutrient solution transport pipe (11), at least one liquid droping port (12) forming for placing the horizontal support bar (14) of shallow-layer culture plate (2); Described illumination apparatus (3) is arranged on the below of any one horizontal support bar (14) of Multilayer culture shelf (1), described Multilayer culture shelf (1) bottom is provided with nutrient solution recovering tray (4), and described nutrient solution recovering tray (4) is communicated with nutrient solution pond for recovering (7) by recovery channel (5), recovery liquid filtering system (6); Described nutrient solution pond for recovering (7) is provided with nutrient solution transferpump (8) and temperature control device (9) in being communicated with; Described nutrient solution transferpump (8) is communicated with liquid droping port (12) by nutrient solution transport pipe (11), and described liquid droping port (12) is positioned at the top of Multilayer culture shelf (1) top layer; Described shallow-layer culture plate (2) bottom is provided with multiple holes, is equipped with sealing plug (15) in described hole, is provided with a Height Adjustable liquid level control minim pipette (16) in the middle of described sealing plug (15).
2. Nostoc algae shallow-layer culture systems according to claim 1, is characterized in that, described nutrient solution transport pipe (11) is provided with nutrient solution delivery controller (10).
3. Nostoc algae shallow-layer culture systems according to claim 1, is characterized in that, the aperture of described shallow-layer culture plate (2) bottom is 0.8-1.2 cm.
4. Nostoc algae shallow-layer culture systems according to claim 1, it is characterized in that, the caliber of described liquid level control minim pipette (16) is 1mm-2 mm, pipe range 4cm-8cm, and the caliber of described liquid level control minim pipette (16) is less than the diameter of cultivated algae kind.
5. Nostoc algae shallow-layer culture systems according to claim 1, is characterized in that, the interlamellar spacing of described Multilayer culture shelf (1) is 30 cm-40cm, and interlamellar spacing is adjustable.
6. Nostoc algae shallow-layer culture systems according to claim 1, is characterized in that, described liquid droping port (12) is 40cm-50cm with the distance of Multilayer culture shelf (1) top layer.
7. Nostoc algae shallow-layer cultural method, is characterized in that, right to use requires one of 1-6 described culture systems, and concrete steps are:
(1) take glasshouse or plastic greenhouse greenhouse as cultivating place, described one of claim 1-6 culture systems is placed in and is cultivated in place, shallow-layer culture plate is placed on Multilayer culture shelf, the every interlayer of Multilayer culture shelf is divided into 30 cm～40cm, and the distance that liquid droping port and Multilayer culture shelf top layer are set is 40cm-50cm; Described shallow-layer culture plate bottom is provided with multiple holes, and shallow-layer culture plate aperture is 0.8-1.2 cm, in described hole, is equipped with sealing plug, is provided with a Height Adjustable liquid level control minim pipette in the middle of described sealing plug;
(2) inoculation is the algae kind that the diameter that obtains by sterile culture is 1mm～2mm by algae kind, when inoculation, algae kind is evenly laid in shallow-layer culture plate, and the density of inoculation is controlled at 0.5 g/cm
2～1g/cm
2, inoculation thickness is 1 cm-2cm;
(3) select diameter to be less than the liquid level control minim pipette of Nostoc particle diameter according to the diameter of culture, regulate liquid level control minim pipette height on culture plate, make the nutrient solution liquid level drenching into culture plate just flood culture, unnecessary nutrient solution flows into lower floor's culture plate by liquid level control minim pipette, automatically dissolves in fresh air when nutrient solution splashes into culture plate; Meanwhile, controlled light remains between 1000lx～20000lx, controls culture-liquid temp and remains between 20 ℃～30 ℃;
Described nutrient solution composition and concentration are:
MgSO
4. 7H
2O 0.05-0.15g/L
K
2HPO
4. 3H
2O 0.005-0.08g/L
CaCl
2. 2H
2O 0.004-0.06g/L
EDTA-Na
2 0.001-0.004g/ L
Ferric ammonium citrate 0.001-0.005 g/ L
H
3BO
4 0.002-0.008mg/L
Na
2MoO
4.2H
2O 0.001-0.030mg/L
ZnSO
4.7H
2O 0.001-0.02mg/L
CuSO
4.5H
2O 0.001-0.005mg/L
MnCl
2.4H
2O 0.020-0.060mg/L。
8. Nostoc algae shallow-layer cultural method according to claim 7, is characterized in that, described Nostoc algae is Nostoc or Nostoc commune.
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