CN113817611A - Culture medium and autotrophic culture method for desert Chlorella foenum-graecum - Google Patents
Culture medium and autotrophic culture method for desert Chlorella foenum-graecum Download PDFInfo
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- 239000001963 growth medium Substances 0.000 title claims abstract description 32
- 241000195649 Chlorella <Chlorellales> Species 0.000 title claims abstract description 13
- 230000001651 autotrophic effect Effects 0.000 title claims abstract description 12
- 238000012136 culture method Methods 0.000 title claims abstract description 11
- 241000195654 Chlorella sorokiniana Species 0.000 claims abstract description 66
- NWBJYWHLCVSVIJ-UHFFFAOYSA-N N-benzyladenine Chemical compound N=1C=NC=2NC=NC=2C=1NCC1=CC=CC=C1 NWBJYWHLCVSVIJ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 239000005631 2,4-Dichlorophenoxyacetic acid Substances 0.000 claims abstract description 8
- 238000012258 culturing Methods 0.000 claims abstract description 5
- 229930195732 phytohormone Natural products 0.000 claims abstract description 4
- KRKNYBCHXYNGOX-UHFFFAOYSA-N citric acid Natural products OC(=O)CC(O)(C(O)=O)CC(O)=O KRKNYBCHXYNGOX-UHFFFAOYSA-N 0.000 claims description 25
- VWDWKYIASSYTQR-UHFFFAOYSA-N sodium nitrate Chemical compound [Na+].[O-][N+]([O-])=O VWDWKYIASSYTQR-UHFFFAOYSA-N 0.000 claims description 18
- 238000005286 illumination Methods 0.000 claims description 10
- CDBYLPFSWZWCQE-UHFFFAOYSA-L sodium carbonate Substances [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 10
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 9
- 239000011573 trace mineral Substances 0.000 claims description 9
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 8
- 239000001110 calcium chloride Substances 0.000 claims description 8
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 8
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 8
- 229910000396 dipotassium phosphate Inorganic materials 0.000 claims description 8
- 229910052564 epsomite Inorganic materials 0.000 claims description 8
- XLYOFNOQVPJJNP-ZSJDYOACSA-N heavy water Substances [2H]O[2H] XLYOFNOQVPJJNP-ZSJDYOACSA-N 0.000 claims description 8
- 235000013619 trace mineral Nutrition 0.000 claims description 8
- 239000002609 medium Substances 0.000 claims description 3
- 239000002994 raw material Substances 0.000 claims description 2
- PRPINYUDVPFIRX-UHFFFAOYSA-N 1-naphthaleneacetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CC=CC2=C1 PRPINYUDVPFIRX-UHFFFAOYSA-N 0.000 claims 3
- 239000005972 6-Benzyladenine Substances 0.000 claims 3
- JTEDVYBZBROSJT-UHFFFAOYSA-N indole-3-butyric acid Chemical group C1=CC=C2C(CCCC(=O)O)=CNC2=C1 JTEDVYBZBROSJT-UHFFFAOYSA-N 0.000 claims 3
- PXRKCOCTEMYUEG-UHFFFAOYSA-N 5-aminoisoindole-1,3-dione Chemical compound NC1=CC=C2C(=O)NC(=O)C2=C1 PXRKCOCTEMYUEG-UHFFFAOYSA-N 0.000 claims 2
- FRHBOQMZUOWXQL-UHFFFAOYSA-L ammonium ferric citrate Chemical compound [NH4+].[Fe+3].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O FRHBOQMZUOWXQL-UHFFFAOYSA-L 0.000 claims 2
- 229960004642 ferric ammonium citrate Drugs 0.000 claims 2
- 239000004313 iron ammonium citrate Substances 0.000 claims 2
- 235000000011 iron ammonium citrate Nutrition 0.000 claims 2
- 239000002054 inoculum Substances 0.000 claims 1
- 239000003375 plant hormone Substances 0.000 claims 1
- 238000000034 method Methods 0.000 abstract description 6
- 241000195493 Cryptophyta Species 0.000 abstract description 2
- 230000005526 G1 to G0 transition Effects 0.000 abstract description 2
- 230000009286 beneficial effect Effects 0.000 abstract description 2
- 239000000843 powder Substances 0.000 abstract 1
- 238000001035 drying Methods 0.000 description 10
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 6
- 238000005273 aeration Methods 0.000 description 5
- 238000001816 cooling Methods 0.000 description 5
- 238000010438 heat treatment Methods 0.000 description 5
- 238000005259 measurement Methods 0.000 description 5
- 239000012528 membrane Substances 0.000 description 5
- 230000001954 sterilising effect Effects 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 238000005303 weighing Methods 0.000 description 5
- 239000008280 blood Substances 0.000 description 4
- 210000004369 blood Anatomy 0.000 description 4
- 230000003203 everyday effect Effects 0.000 description 4
- 230000003287 optical effect Effects 0.000 description 4
- 240000009108 Chlorella vulgaris Species 0.000 description 2
- 235000007089 Chlorella vulgaris Nutrition 0.000 description 2
- 244000077995 Coix lacryma jobi Species 0.000 description 2
- 244000062793 Sorghum vulgare Species 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 235000019713 millet Nutrition 0.000 description 2
- 241000894007 species Species 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 1
- 241000721153 Chloranthus japonicus Species 0.000 description 1
- -1 EDTANa2 Chemical compound 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 238000004891 communication Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 150000002484 inorganic compounds Chemical class 0.000 description 1
- 229910010272 inorganic material Inorganic materials 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 238000003825 pressing Methods 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 235000017550 sodium carbonate Nutrition 0.000 description 1
- 235000010344 sodium nitrate Nutrition 0.000 description 1
- YWYZEGXAUVWDED-UHFFFAOYSA-N triammonium citrate Chemical compound [NH4+].[NH4+].[NH4+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O YWYZEGXAUVWDED-UHFFFAOYSA-N 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- 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
- C12N1/12—Unicellular algae; Culture media therefor
-
- C—CHEMISTRY; METALLURGY
- 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
- C12N1/38—Chemical stimulation of growth or activity by addition of chemical compounds which are not essential growth factors; Stimulation of growth by removal of a chemical compound
Abstract
The invention discloses a desert Chlorella foenum-graecum culture medium and an autotrophic culture method. The culture medium for the rapid growth of the desert Chlorella sorokiniana is prepared by adding four different phytohormones (IBA, 2,4-D, NAA, 6-BA) and the like into a conventional culture solution for the growth of the desert Chlorella sorokiniana, and then culturing the desert Chlorella sorokiniana. Compared with the prior art, the invention has the beneficial effects that: when the cultured desert Chlorella sorokiniana cells reach a stationary phase, 1.5-3.5 g of cell dry powder can be obtained by each liter of culture solution, which is 100% higher than that of the conventional culture method for culturing desert Chlorella sorokiniana, so that the yield of desert algae is effectively improved, the culture cost is reduced, and the method has wide application prospect.
Description
Technical Field
The invention relates to the technical field of Saint Wu, in particular to a culture medium and an autotrophic culture method for Chlorella foenum graecum in desert.
Background
The desert Chlorella sorokiniana (Chlorella sorokiniana) is a species published in 1965, and compared with the Chlorella vulgaris (Chlorella vulgaris) published in 1890, the Chlorella sorokiniana has the characteristics of faster growth, high temperature resistance and pollution resistance, and is particularly easy to be dominant in outdoor large-scale culture. The species can be widely applied to some recent energy microalgae or microalgae biotechnology related documents at home and abroad.
Chlorella is the first to be applied to commercial scale production, is one of the more algae species developed at present, has easy culture, is rich in oil and fat such as oil containing unsaturated fatty acid and protein in cells, and has good application prospect in the fields of biological materials, nutritional foods and health care products
At present, the culture mode of the chlorella sorokiniana in the desert can be divided into autotrophic culture and heterotrophic culture. The autotrophic culture of the desert Chlorella sorokiniana can utilize a cold light source LED illuminating lamp, and a culture medium mainly comprises inorganic compounds. At present, the autotrophic culture technology of the chlorella sorokiniana in desert is basically mature, but the culture method of rapidly dividing cells in a short period to increase the cell density limits the further increase of the biomass.
Disclosure of Invention
The invention aims to provide a desert Chlorella foenum-graecum culture medium and an autotrophic culture method, so as to solve the problems in the background technology.
In order to achieve the aim, the invention provides the following technical scheme that the raw materials of the culture medium for the chloranthus japonicus comprise NaNO3, K2HPO 4.3H 2O, MgSO 4.7H 2O, CaCl 2.2H 2O, citric acid, ferri ammonium citrate, EDTANa2, Na2CO3 and trace elements; at a concentration of NaNO3 1.5g/L、K2HPO4·3H2O 40mg/L、MgSO4·7H2O 75mg/L、CaCl2·2H2O 36mg/L、citric acid 6mg/L、ferric ammonium citrate6mg/L、EDTANa2 1mg/L、Na2CO320mg/L, 1m of trace element solution and 5-20mg/L of phytohormone.
An autotrophic culture method of Chlorella foenum graecum in desert comprises the following culture steps: placing Chlorella sorokiniana into the culture medium of claim 1, culturing at 27-31 deg.C and illumination intensity of 2500-.
As a preferable technical scheme of the invention, in the step one, a mixture of the coix seed and the millet can be selected, and the proportion of the coix seed to the millet is not lower than 3: 1.
As a preferred technical scheme of the invention, the light-dark ratio of each day during culture is 12 h: 12h to 20h: and 4 h.
As a preferred embodiment of the present invention, the initial inoculation amount in the medium during the cultivation is 1X 104-1×105cell/mL。
Compared with the prior art, the invention has the beneficial effects that: when the cultured Chlorella sorokiniana cells reach the stationary phase, 1.5-3.5 g of dry cell matter can be obtained by each liter of culture solution, which is 100% higher than that of the conventional culture solution of the Chlorella sorokiniana without phytohormones (IBA, 2,4-D, NAA, 6-BA), thereby effectively improving the yield of the Chlorella sorokiniana cells, reducing the culture cost and having wider application prospect.
Drawings
FIG. 1 is a graph showing the growth of Chlorella sorokiniana in a desert after culturing according to the present invention;
Detailed Description
Example 1
Taking the concentration as NaNO3 1.5g/L、K2HPO4·3H2O 40mg/L、MgSO4·7H2O 75mg/L、CaCl2·2H2O 36mg/L、citric acid 6mg/L、ferric ammonium citrate6mg/L、EDTANa2 1mg/L、Na2CO320mg/L, 1mL of trace element solution, adding pure water to a constant volume of 1L, heating to 121 ℃ and sterilizing for 20min to obtain the conventional culture solution.
As a preferable embodiment of the present invention, IBA, 2,4-D, NAA and 6-BA each in an amount of 5mg/L are added to a conventional culture medium to obtain a fast-growing culture medium.
As a preferred technical scheme of the invention, under the aseptic condition, the activated Chlorella sorokiniana cells in desert are inoculated into a culture medium of a culture solution for the rapid growth of the Chlorella sorokiniana in desert, and the initial concentration of the cells is 1 x 104cell/mL。
As a preferred technical scheme of the invention, the culture medium is placed in an illumination culture frame for aeration culture, the culture temperature is 27 ℃, the illumination intensity is 2500lux, and the light-dark ratio is 20h:4 h.
As a preferred technical scheme of the invention, the growth condition of the cells of the chlorella sorokiniana in the desert is determined as follows: during the culture period, 2mL of culture solution is taken from the culture bottle every day, the cells of the Chlorella sorokiniana in desert are counted by a blood counting plate or the optical density value of the culture solution at the wavelength of 680nm is measured by a spectrophotometer, and the growth stage of the cells of the Chlorella sorokiniana in desert is determinedWhen the cell number or OD of Chlorella sorokiniana in desert680The culture is completed when the culture is stable.
As a preferred technical scheme of the invention, the method comprises the following steps of collecting desert Chlorella sorokiniana cells: 50mL of the culture solution of Chlorella sorokiniana cells in which the culture was completed was taken, and the cells were collected through a vacuum filter using a 0.22um filter for measurement of the dry weight of the cells.
Collecting dry weight of cells of Chlorella sorokiniana in desert: and (3) drying the obtained filter membrane collected with the desert Chlorella sorokiniana cells in an oven at 105 ℃ for 4 hours, weighing and calculating after drying and cooling to obtain the cell dry matter content of 1.5g/L in the culture solution in unit volume.
Example 2
Taking the concentration as NaNO3 1.5g/L、K2HPO4·3H2O 40mg/L、MgSO4·7H2O 75mg/L、CaCl2·2H2O 36mg/L、citric acid 6mg/L、ferric ammonium citrate6mg/L、EDTANa2 1mg/L、Na2CO320mg/L, 1mL of trace element solution, adding pure water to a constant volume of 1L, heating to 121 ℃ and sterilizing for 20min to obtain the conventional culture solution.
As a preferable embodiment of the present invention, IBA, 2,4-D, NAA and 6-BA each in an amount of 9mg/L are added to a conventional culture medium to obtain a fast-growing culture medium.
As a preferred technical scheme of the invention, under the aseptic condition, the activated Chlorella sorokiniana cells in desert are inoculated into a culture medium of a culture solution for the rapid growth of the Chlorella sorokiniana in desert, and the initial concentration of the cells is 1 x 105cell/mL。
As a preferred technical scheme of the invention, the culture medium is placed in an illumination culture frame for aeration culture, the culture temperature is 28 ℃, the illumination intensity is 3000lux, and the light-dark ratio is 14h:10 h.
As a preferred technical scheme of the invention, the growth condition of the cells of the chlorella sorokiniana in the desert is determined as follows: during the culture period, 2mL of culture solution is taken from the culture flask every day, and cells of Chlorella sorokiniana in desert are counted by using a blood counting plate or the wavelength of the culture solution at 680nm is measured by using a spectrophotometerDetermining the growth stage of Chlorella sorokiniana cells in desert, when the number or OD of the Chlorella sorokiniana cells in desert680The culture is completed when the culture is stable.
As a preferred technical scheme of the invention, the method comprises the following steps of collecting desert Chlorella sorokiniana cells: 50mL of the culture solution of Chlorella sorokiniana cells in which the culture was completed was taken, and the cells were collected through a vacuum filter using a 0.22um filter for measurement of the dry weight of the cells.
Collecting dry weight of cells of Chlorella sorokiniana in desert: and (3) drying the obtained filter membrane collected with the desert Chlorella sorokiniana cells in an oven at 105 ℃ for 4 hours, weighing and calculating after drying and cooling to obtain the cell dry matter content of 1.9g/L in the culture solution in unit volume.
Example 3
Taking the concentration as NaNO3 1.5g/L、K2HPO4·3H2O 40mg/L、MgSO4·7H2O 75mg/L、CaCl2·2H2O 36mg/L、citric acid 6mg/L、ferric ammonium citrate6mg/L、EDTANa2 1mg/L、Na2CO320mg/L, 1mL of trace element solution, adding pure water to a constant volume of 1L, heating to 121 ℃ and sterilizing for 20min to obtain the conventional culture solution.
As a preferable embodiment of the present invention, a fast-growing culture solution is obtained by adding 13mg/L each of IBA, 2,4-D, NAA, and 6-BA to a conventional culture solution.
As a preferred technical scheme of the invention, under the aseptic condition, the activated Chlorella sorokiniana cells in desert are inoculated into a culture medium of a culture solution for the rapid growth of the Chlorella sorokiniana in desert, and the initial concentration of the cells is 1 x 104cell/mL。
As a preferred technical scheme of the invention, the culture medium is placed in an illumination culture frame for aeration culture, the culture temperature is 29 ℃, the illumination intensity is 3500lux, and the light-dark ratio is 16h:8 h.
As a preferred technical scheme of the invention, the growth condition of the cells of the chlorella sorokiniana in the desert is determined as follows: during the culture period, 2mL of culture medium was taken from the culture flask every day and applied to Chlorella sorokiniana in desert with a hemocytometerCounting cells or measuring optical density value of the culture solution at 680nm with spectrophotometer to determine the growth stage of Chlorella sorokiniana cells in desert, when the number or OD of Chlorella sorokiniana cells in desert680The culture is completed when the culture is stable.
As a preferred technical scheme of the invention, the method comprises the following steps of collecting desert Chlorella sorokiniana cells: 50mL of the culture solution of Chlorella sorokiniana cells in which the culture was completed was taken, and the cells were collected through a vacuum filter using a 0.22um filter for measurement of the dry weight of the cells.
Collecting dry weight of cells of Chlorella sorokiniana in desert: and (3) drying the obtained filter membrane collected with the desert Chlorella sorokiniana cells in an oven at 105 ℃ for 4 hours, weighing and calculating after drying and cooling to obtain the cell dry matter content of 2.5g/L in the culture solution in unit volume.
Example 4
Taking the concentration as NaNO3 1.5g/L、K2HPO4·3H2O 40mg/L、MgSO4·7H2O 75mg/L、CaCl2·2H2O 36mg/L、citric acid 6mg/L、ferric ammonium citrate6mg/L、EDTANa2 1mg/L、Na2CO320mg/L, 1mL of trace element solution, adding pure water to a constant volume of 1L, heating to 121 ℃ and sterilizing for 20min to obtain the conventional culture solution.
As a preferable embodiment of the present invention, IBA, 2,4-D, NAA and 6-BA each in an amount of 17mg/L is added to a conventional culture medium to obtain a fast-growing culture medium.
As a preferred technical scheme of the invention, under the aseptic condition, the activated Chlorella sorokiniana cells in desert are inoculated into a culture medium of a culture solution for the rapid growth of the Chlorella sorokiniana in desert, and the initial concentration of the cells is 1 x 105cell/mL。
As a preferred technical scheme of the invention, the culture medium is placed in an illumination culture frame for aeration culture, the culture temperature is 29 ℃, the illumination intensity is 4000lux, and the light-dark ratio is 18h:6 h.
As a preferred technical scheme of the invention, the growth condition of the cells of the chlorella sorokiniana in the desert is determined as follows: during the culture period, the culture medium is cultured every dayTaking 2mL of culture solution from the bottle, counting cells of Chlorella sorokiniana in desert by using a blood counting plate or measuring optical density value of the culture solution at 680nm by using a spectrophotometer to determine the growth stage of the cells of Chlorella sorokiniana in desert, and determining the number or OD of the cells of the Chlorella sorokiniana in desert680The culture is completed when the culture is stable.
As a preferred technical scheme of the invention, the method comprises the following steps of collecting desert Chlorella sorokiniana cells: 50mL of the culture solution of Chlorella sorokiniana cells in which the culture was completed was taken, and the cells were collected through a vacuum filter using a 0.22um filter for measurement of the dry weight of the cells.
Collecting dry weight of cells of Chlorella sorokiniana in desert: and (3) drying the obtained filter membrane collected with the desert Chlorella sorokiniana cells in an oven at 105 ℃ for 4h, weighing after drying and cooling, and calculating to obtain the cell dry matter content in the culture solution of unit volume of 3 g/L.
Example 5
Taking the concentration as NaNO3 1.5g/L、K2HPO4·3H2O 40mg/L、MgSO4·7H2O 75mg/L、CaCl2·2H2O 36mg/L、citric acid 6mg/L、ferric ammonium citrate6mg/L、EDTANa2 1mg/L、Na2CO320mg/L, 1mL of trace element solution, adding pure water to a constant volume of 1L, heating to 121 ℃ and sterilizing for 20min to obtain the conventional culture solution.
As a preferable embodiment of the present invention, IBA, 2,4-D, NAA and 6-BA each in an amount of 20mg/L are added to a conventional culture medium to obtain a fast-growing culture medium.
As a preferred technical scheme of the invention, under the aseptic condition, the activated Chlorella sorokiniana cells in desert are inoculated into a culture medium of a culture solution for the rapid growth of the Chlorella sorokiniana in desert, and the initial concentration of the cells is 1 x 105cell/mL。
As a preferred technical scheme of the invention, the culture medium is placed in a light culture frame for aeration culture, the culture temperature is 30 ℃, the light intensity is 4500lux, and the light-dark ratio is 20h:4 h.
As a preferred technical scheme of the invention, the desert Solomon is smallDetermination of growth of chlorella cells: during the culture period, 2mL of culture solution is taken from the culture bottle every day, the cells of the Chlorella sorokiniana in desert are counted by a blood counting plate or the optical density value of the culture solution at the wavelength of 680nm is measured by a spectrophotometer, the growth stage of the cells of the Chlorella sorokiniana in desert is determined, and when the number or OD of the cells of the Chlorella sorokiniana in desert is reached680The culture is completed when the culture is stable.
As a preferred technical scheme of the invention, the method comprises the following steps of collecting desert Chlorella sorokiniana cells: 50mL of the culture solution of Chlorella sorokiniana cells in which the culture was completed was taken, and the cells were collected through a vacuum filter using a 0.22um filter for measurement of the dry weight of the cells.
Collecting dry weight of cells of Chlorella sorokiniana in desert: and (3) drying the obtained filter membrane collected with the desert Chlorella sorokiniana cells in an oven at 105 ℃ for 4h, weighing after drying and cooling, and calculating to obtain the cell dry matter content in the culture solution of unit volume of 3.5 g/L.
As a preferable technical scheme of the invention, the pressing forming in the fourth step can be square
In the description of the present invention, it is to be understood that the terms "center", "longitudinal", "lateral", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc., indicate orientations or positional relationships based on those shown in the drawings, and are used only for convenience in describing and simplifying the description, but do not indicate or imply that the devices or elements referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore are not to be construed as limiting the invention, and further, the terms "first", "second", etc., are used only for descriptive purposes and are not intended to indicate or imply relative importance or to implicitly indicate the number of technical features indicated, whereby the features defined as "first", "second", etc., may explicitly or implicitly include one or more of such features, in the description of the present invention, "a plurality" means two or more unless otherwise specified.
In the description of the present invention, it should be noted that, unless otherwise explicitly specified or limited, the terms "mounted," "connected," and "connected" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements, and the specific meaning of the above terms in the present invention can be understood by those skilled in the art through specific situations
Although the present invention has been described in detail with reference to the specific embodiments, the present invention is not limited to the above embodiments, and various changes and modifications without inventive changes may be made within the knowledge of those skilled in the art without departing from the spirit of the present invention.
Claims (5)
1. A desert Chlorella foeniculiformis culture medium is characterized in that: the raw material comprises NaNO3、K2HPO4·3H2O、MgSO4·7H2O、CaCl2·2H2O、citric acid、ferric ammonium citrate、EDTANa2、Na2CO3Trace elements; at a concentration of NaNO3 1.5g/L、K2HPO4·3H2O 40mg/L、MgSO4·7H2O 75mg/L、CaCl2·2H2O 36mg/L、citric acid 6mg/L、ferric ammonium citrate 6mg/L、EDTANa2 1mg/L、Na2CO320mg/L, 1mL of trace element solution and 5-20mg/L of phytohormone.
2. The culture medium for Chlorella foeniculiformis according to claim 1, which comprises: the plant hormone is indolebutyric acid (IBA), 2,4-D, naphthylacetic acid (NAA) and 6-benzyladenine (6-BA); the concentration ratio IBA: 2, 4-D: NAA: 6-BA is 1:1:1: 1.
3. An autotrophic culture method of Chlorella foenum graecum in desert is characterized by comprising the following culture steps: placing the chlorella sorokiniana into the culture medium of claim 1, culturing for 7-14 days under the conditions that the temperature is controlled to be 27-31 ℃ and the illumination intensity is 2500-4500 Lux.
4. The autotrophic culture method of Chlorella foeniculiformis in desert according to claim 3, characterized in that: the daily light-dark ratio in the culture is 12 h: 12h to 20h: and 4 h.
5. The autotrophic culture method of Chlorella foeniculiformis in desert according to claim 4, wherein: the initial inoculum size in the medium during culture was 1X 104-1×105cell/mL。
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