CN114196547B - Application of DCMU (dendritic cell activator-terminator) in improving chlorophyll yield or strong light tolerance during microalgae polyculture fermentation - Google Patents

Application of DCMU (dendritic cell activator-terminator) in improving chlorophyll yield or strong light tolerance during microalgae polyculture fermentation Download PDF

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CN114196547B
CN114196547B CN202210139373.1A CN202210139373A CN114196547B CN 114196547 B CN114196547 B CN 114196547B CN 202210139373 A CN202210139373 A CN 202210139373A CN 114196547 B CN114196547 B CN 114196547B
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刘芳华
谢章彰
王欧美
汤佳
郝钦钦
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Institute of Eco Environmental and Soil Sciences of Guangdong Academy of Sciens
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Abstract

The invention discloses application of DCMU (dendritic cell activator-associated monomer) in improving chlorophyll yield or strong light tolerance during microalgae polyculture fermentation. The inventor of the invention discovers that DCMU has the functions of improving the strong light tolerance of microalgae and improving the chlorophyll yield of the microalgae polyculture fermentation in the microalgae polyculture fermentation for the first time, thereby providing a method for improving the strong light tolerance and the chlorophyll yield of the microalgae. Because DCMU is very cheap and has extremely low use concentration, almost no additional industrial cost is generated, the method is very suitable for the industrial culture of microalgae; the invention has the characteristics of simple and convenient operation and strong practicability, and is beneficial to large-scale industrial application.

Description

Application of DCMU (dendritic cell activator-terminator) in improving chlorophyll yield or strong light tolerance during microalgae polyculture fermentation
Technical Field
The invention belongs to the technical field of microalgae biology, and particularly relates to application of DCMU (dendritic cell activator-associated monomer) in improving chlorophyll yield or strong light tolerance during microalgae polyculture fermentation.
Background
Microalgae are a generic term for a class of unicellular photosynthetic microorganisms. Microalgae have become an attractive alternative to traditional terrestrial plants for the production of chlorophyll, biofuels and other high value products, while relieving pressure on arable land, carbon dioxide emissions and global warming. However, achieving high productivity of microalgae products remains a major challenge for commercial application of microalgae. The mixed culture fermentation of the microalgae refers to a culture mode in which the microalgae grows by utilizing organic matters and illumination at the same time, and has the advantages of photoautotrophic growth and heterotrophic growth, the mixed culture fermentation of the microalgae can become a method for improving the growth speed and the product yield of the microalgae, and the microalgae such as chlorella, chlamydomonas, nannochloropsis, scenedesmus obliquus and the like can be found to grow in a mixed culture mode. The advantages of the mixed culture fermentation technology in the aspects of producing biodiesel, biological products and the like by microalgae have been widely researched. However, when microalgae are grown by using natural light, the light intensity of the light is too strong (such as at noon) due to the non-adjustable natural light, and the light damage is generated to the microalgae, so that the growth speed of the microalgae is inhibited. Although the illumination control methods such as an artificial illumination control system and artificial shading can relieve the damage of over-strong illumination to the microalgae to a certain extent and improve the growth speed of the microalgae, the illumination control methods face the problems of high cost, complex operation and the like, and cannot be applied on an industrial level. Therefore, it is necessary to find a new method for improving the tolerance of microalgae to high light intensity with low cost and high efficiency.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provide the application of DCMU in improving the chlorophyll yield or strong light tolerance of microalgae polyculture fermentation.
The purpose of the invention is realized by the following technical scheme: the application of DCMU (dichlorophenyl dimethyl urea) in improving the yield of chlorophyll or the strong light tolerance of microalgae in mixed culture fermentation is based on the research result of finding that DCMU has the effect of improving the yield of chlorophyll or the strong light tolerance of microalgae in mixed culture fermentation of microalgae by the inventor of the invention.
The application of the DCMU in improving the chlorophyll yield or the strong light tolerance during the microalgae polyculture fermentation specifically comprises the following steps: inoculating the microalgae into a mixed culture medium containing DCMU for light culture.
The microalgae is capable of mixed culture and growth, and is preferably chlorella, chlamydomonas, nannochloropsis or scenedesmus obliquus.
The chlorella is preferably Chlorella keshii.
The microalgae is preferably microalgae obtained by enrichment culture of an autotrophic culture medium.
The autotrophic culture medium is preferably BG11 culture medium.
The inoculation amount of the microalgae is preferably calculated according to the volume percentage of 1-3% of the mixed culture medium.
The microalgae is preferably of a cell density of 1X 108～1×109one/mL of microalgae.
The concentration of the DCMU in the mixed culture medium is 20-100 mug/L; preferably 40-80 mug/L; more preferably 60 to 80. mu.g/L.
The mixed culture medium is a culture medium containing a carbon source.
The carbon source is preferably glucose.
The culture medium is preferably BG11 culture medium.
The conditions for the light culture are preferably as follows: the temperature is 25-35 ℃, and the illumination intensity is 2000-12000 Lux; more preferably as follows: the temperature is 30 ℃, and the illumination intensity is 4000-12000 Lux.
The period of the illumination culture is preferably 16-24 h of light/0-8 h of darkness; more preferably 16h light/8 h dark period.
The time of the light culture is preferably more than 24 hours; more preferably 24 to 72 hours.
Compared with the prior art, the invention has the following advantages and effects:
(1) DCMU (dichlorophenyl dimethylurea) is a common herbicide that inhibits the photosynthesis of green plants by inhibiting the electron transfer of PS ii in the photosynthetic system, thereby inhibiting the growth of microalgae. The inventor of the invention discovers that DCMU has the functions of improving the microalgae highlight tolerance and chlorophyll content of the microalgae for mixotrophic fermentation, so that a method is provided for improving the microalgae highlight tolerance and chlorophyll content of the microalgae.
(2) Because DCMU is very cheap and has extremely low use concentration, and hardly generates additional industrial cost, the method provided by the invention is very suitable for the industrial culture of microalgae.
(3) The method provided by the invention has the characteristics of simple and convenient operation and strong practicability, and is beneficial to large-scale industrial application.
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FIG. 1 is a graph of the results of the effect of DCMU at various concentrations on the autotrophic and mixotrophic growth of Chlorella Kelvin: p < 0.01.
FIG. 2 is a graph showing the effect of DCMU at various concentrations on the growth of Chlorella Kelvin by mixotrophic cultivation.
FIG. 3 is a graph showing the effect of DCMU and different chlorophyll contents on the growth of C.kichenensis; wherein A is the change result of chlorophyll content and biomass of different experimental groups cultured for 12 hours, and B is the detection result of microalgae glucose absorption and microalgae biomass of different polyculture experimental groups cultured for 1 hour; p < 0.01.
FIG. 4 is a graph showing the effect of DCMU on the mixotrophic growth of Chlorella Kelvin under different light intensities; CK is a control group, and DCMU is an experimental group added with DCMU. P < 0.05, P < 0.01.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
Example 1:
(1) chlorella vulgaris (C. Kelvin)Parachlorella kessleri) FACHB-4 (purchased from fresh water algae seed bank of Chinese academy of sciences) is used as microalgae seed, and is first subjected to enrichment culture by using BG11 culture solution until the cell density of the Chlorella Kelly reaches 1 × 108～1×109When the seeds are cultured per mL, the Chlorella keiskei seed liquid is obtained. BG11 culture solution comprises the following components: NaNO3 1.5 g/L、K2HPO4 40 mg/L、MgSO4·7H2O 70 mg/L，CaCl2·2H2O40 mg/L, citric acid 6 mg/L, ferric ammonium citrate 6 mg/L, EDTA-Na2 1 mg/L、Na2CO3 20 mg/L、H3BO3 2.86 mg/L、MnCl2·4H2O 1.86 mg/L、ZnSO4·7H2O 0.22 mg/L、Na2MoO4·2H2O 0.39 mg/L、CuSO4·5H2O 0.08 mg/L、Co(NO3)2·6H2O 0.05 mg/L，pH＝7.00。
(2) Preparing a microalgae glucose polyculture medium, wherein the culture medium is BG11 culture medium plus 2 g/L glucose and 2 g/L Tris base (Tris-base), and the pH value is 7.00.
(3) Inoculating the chlorella keiskei seed solution into a microalgae glucose mixed culture medium and an autotrophic BG11 culture medium according to the volume ratio of 2%, adding DCMU (0, 5, 10, 20, 40, 80, 120, 160, 200 and 300 mu g/L) with different final concentrations, and culturing the chlorella keiskei culture in a light incubator under the culture conditions that: the biomass (dry weight) was measured by culturing at 30 ℃ under 6000 Lux illumination for 16/8 hours with an illumination cycle for 3 days. The results are shown in FIG. 1:
A. DCMU inhibits autotrophic growth of Chlorella Kelvin, and the inhibition ability increases with the increase of the addition amount of DCMU, and when the addition amount of DCMU is above 160 μ g/L, the DCMU has lethal effect on Chlorella Kelvin.
B. When the chlorella vulgaris is kept in a mixed state for a long time, the growth rate of the chlorella vulgaris tends to increase firstly and then decrease with the increase of the addition amount of DCMU. When the addition amount of DCMU is 20-80 mug/L, the growth of the Chlorella Kelvin is remarkably promoted, and when the addition amount of DCMU is more than 80 mug/L, the mixotrophic growth of the Chlorella Kelvin is inhibited.
It can be seen that DCMU has different effects on the autotrophic and mixotrophic growth of Chlorella Kelvin.
Method for measuring dry weight: firstly, microalgae OD is established680And the standard curve between the dry weight of microalgae and the dry weight of microalgae is obtained by measuring the OD of microalgae culture solution680And (5) converting the dry weight of the microalgae. In the examples, Chlorella Kelvin: (Parachlorella kessleri) FACHB-4 dry weight standard: dry weight (g/L) =0.423 × OD680-0.005。
Example 2:
(1) chlorella vulgaris (C. Kelvin)Parachlorella kessleri) FACHB-4 (purchased from fresh water algae seed bank of Chinese academy of sciences) is used as microalgae seed, and is first subjected to enrichment culture by using BG11 culture solution until the cell density of the Chlorella Kelly reaches 1 × 108～1×109When the seeds are cultured per mL, the Chlorella keiskei seed liquid is obtained. BG11 culture solution comprises the following components: NaNO3 1.5 g/L、K2HPO4 40 mg/L、MgSO4·7H2O 70 mg/L，CaCl2·2H2O40 mg/L, citric acid 6 mg/L, ferric ammonium citrate 6 mg/L, EDTA-Na2 1 mg/L、Na2CO3 20 mg/L、H3BO3 2.86 mg/L、MnCl2·4H2O 1.86 mg/L、ZnSO4·7H2O 0.22 mg/L、Na2MoO4·2H2O 0.39 mg/L、CuSO4·5H2O 0.08 mg/L、Co(NO3)2·6H2O 0.05 mg/L，pH＝7.00。
(2) Preparing a microalgae glucose polyculture medium, wherein the culture medium is BG11 culture medium plus 2 g/L glucose and 2 g/L Tris base (Tris-base), and the pH value is 7.00.
(3) Inoculating the chlorella keiskei seed solution to a microalgae glucose mixed culture medium according to the volume ratio of 2%, adding DCMU (0, 40, 60 and 80 mu g/L) with different final concentrations, and culturing the chlorella keiskei culture in an illumination incubator under the culture conditions: 30 ℃ under 6000 Lux light, 16/8 hours light cycle.
(4) Samples were taken at 12h intervals and the glucose content of the broth, the dry weight of the C.kichenensis and the chlorophyll content were measured, and the results are shown in FIG. 2:
A. after 12 hours of culture, the chlorophyll content of the Chlorella Kelvin is reduced from the initial 12.5 mg/g to 4.97 mg/g, and the reduction range is over 50 percent in the blank control group without addition of DCMU, while the reduction of the chlorophyll content is obviously relieved after addition of DCMU, and the larger the addition amount of DCMU is, the less the chlorophyll content is reduced. When 80. mu.g/L DCMU was added, the content of Chlorella Kelvin was 10.63 mg/g, which was 2.14 times that of the case without DCMU, after 12 hours of cultivation. After 72 hours of culture, the biomass of the chlorella keiskei cultured and grown in the DCMU culture medium containing 60 mu g/L in a mixed way is 2.94 g/L, which is 18.55 percent higher than that of a blank control. And the chlorophyll yield of the chlorella keiskei cultured and grown in the DCMU culture medium containing 60 mu g/L is 49.19 mg/L, which is 81.36% higher than that of the blank control. This result also demonstrates that the addition of DCMU can significantly increase the chlorophyll production and growth rate of C.kichenensis.
B. As the culture medium time passed, the glucose content in the culture solution decreased, and it was found that the Chlorella Kerneyi grew in a polyculture manner using the glucose in the culture solution.
The dry weight was determined as above.
Determination of chlorophyll content: centrifuging 2 mL of microalgae culture solution at 10000 rpm for 2 min, and removing supernatant; adding 100 mu L of distilled water to resuspend the microalgae, adding 1.9 mL of absolute ethyl alcohol, carrying out water bath at 75 ℃ for 5min, centrifuging at 10000 rpm for 2 min, and collecting supernatant; the total supernatant volume was recorded and the absorbance was measured at 665 nm, 649 nm and 750 nm in 95% ethanol as a blank. Leaf based on formulaConversion of chlorophyll content: total chlorophyll (mg/L) Chl = Chla + Chlb =6.10 OD665+20.04 OD649。
The glucose content was determined using the 3, 5-dinitrosalicylic acid method: test solution composition (1L): 6.3 g of 3, 5-dinitrosalicylic acid, 21g of NaOH and KNaC4H12O10·4H2O182 g, phenol 5g, Na2SO35g of the total weight. Diluting the fermentation liquor by 20 times, and mixing the diluted fermentation liquor with detection liquor according to the volume ratio of 1: 3, mixing, carrying out water bath at 100 ℃ for 5min, detecting the absorbance at 540 nm, and calculating according to a standard curve to obtain the glucose content.
Example 3:
(1) chlorella vulgaris (C. Kelvin)Parachlorella kessleri) FACHB-4 is a microalgae species, firstly, using BG11 culture solution to make enrichment culture, when the cell density of the Chlorella Kelvin reaches 1X 108～1×109When the seeds are cultured per mL, the Chlorella keiskei seed liquid is obtained. BG11 culture solution comprises the following components: NaNO3 (1.5 g/L)，K2HPO4 (40 mg/L)，MgSO4·7H2O (70 mg/L)，CaCl2·2H2O (40 mg/L), citric acid (6 mg/L), ferric ammonium citrate (6 mg/L), EDTA-Na2 (1 mg/L)，Na2CO3 (20 mg/L)，H3BO3 (2.86 mg/L)，MnCl2·4H2O (1.86 mg/L)，ZnSO4·7H2O (0.22 mg/L)，Na2MoO4·2H2O (0.39 mg/L)，CuSO4·5H2O (0.08 mg/L)，Co(NO3)2·6H2O (0.05 mg/L)，pH＝7.00。
(2) Preparing microalgae glucose polyculture medium (B + G medium), wherein the culture medium is BG11 medium plus 2G/L glucose and 2G/L Tris base (Tris-base), and the pH value is 7.00.
(3) Inoculating the Kjeldahl quasi-chlorella seed liquid into a microalgae glucose mixed culture medium (B + G) and a microalgae glucose mixed culture medium (B + G + DCMU) containing 60 mu G/L DCMU respectively according to the inoculation amount of 5% (v/v), and culturing 6000 lux for 24 hours to obtain microalgae seeds with low chlorophyll content (microalgae obtained after culturing by the microalgae glucose mixed culture medium) and microalgae seeds with high chlorophyll content (microalgae obtained after culturing by the microalgae glucose mixed culture medium containing 60 mu G/L DCMU).
(4) Microalgae seeds with low chlorophyll content are mixed according to a volume ratio of 1: transferring the culture medium to a new BG11 culture medium according to the proportion of 1 to obtain a low chlorophyll-autotrophic experimental group; microalgae seeds with low chlorophyll content are mixed according to a volume ratio of 1: 1, transferring the microalgae culture medium to a new microalgae glucose polyculture medium in proportion to obtain a low chlorophyll-polyculture experimental group; microalgae seeds with high chlorophyll content are mixed according to a volume ratio of 1: transferring the culture medium to a new BG11 culture medium according to the proportion of 1 to obtain a high chlorophyll-autotrophic experimental group; microalgae seeds with high chlorophyll content are mixed according to a volume ratio of 1: 1 proportion is transferred to BG11 culture medium containing 60 mug/L DCMU to obtain a high chlorophyll-autotrophic-DCMU experimental group; microalgae seeds with high chlorophyll content are mixed according to a volume ratio of 1: 1, transferring the mixture to a microalgae glucose polyculture medium in proportion to obtain a high chlorophyll-polyculture experimental group; microalgae seeds with high chlorophyll content are mixed according to a volume ratio of 1: transferring the mixture to a microalgae glucose mixed culture medium containing 60 mu g/L of DCMU at a ratio of 1 to obtain a high chlorophyll-mixed culture-DCMU experimental group. The aforementioned experimental groups were incubated at 6000 lux at 30 ℃ for 12 hours.
(5) The change in glucose content in the culture system 1 hour after inoculation, the change in dry weight and chlorophyll content of the C.kichenensis before and after culture were measured, and the results are shown in FIG. 3:
A. compared with the Kjeldahl pseudochlorella with low chlorophyll content, the autotrophic growth speed and the mixotrophic growth speed of the Kjeldahl pseudochlorella with high chlorophyll content are obviously improved, which shows that the growth speed of the microalgae can be improved by increasing the chlorophyll content.
B. After 60 mu g/LDCMU is added into the C.kikuii with high chlorophyll content, the autotrophic and polyculture growth rate of the C.kikuii is inhibited to a certain extent. DCMU is a photosynthesis inhibitor, which can inhibit photosynthesis of microalgae, thereby inhibiting autotrophic growth of microalgae, and when the microalgae grow mixedly, the growth speed is a superposition state of autotrophic growth and heterotrophic growth, so that the DCMU can also influence the mixotrophic growth speed of the microalgae to a certain extent.
C. After 60 mu g/LDCMU is added again to the C.trivialis with high chlorophyll content, the autotrophic growth rate of the C.trivialis has no significant difference with the growth rate of the C.trivialis with low chlorophyll content. The result shows that the effect of improving the growth speed of the chlorella keiskei by increasing the chlorophyll content is offset with the effect of inhibiting the autotrophic growth of the chlorella keiskei by DCMU, so that the autotrophic growth speed of the chlorella keiskei with high chlorophyll content is not obviously different from the growth speed of the chlorella keiskei with low chlorophyll content when the chlorella keiskei with high chlorophyll content is grown in a culture medium containing 60 mu g/L of DCMU.
D. After 60 mu g/LDCMU is added again to the Chlorella Kelvin with high chlorophyll content, the mixed culture growth speed of the Chlorella Kelvin with high chlorophyll content is obviously higher than that of the Chlorella Kelvin with low chlorophyll content. Since it is previously known that the autotrophic growth rate of C.kichenii with a high chlorophyll content does not differ significantly from the growth rate of C.kichenii with a low chlorophyll content when it is grown in a medium containing 60. mu.g/LDCMU, the increase in the heterotrophic growth rate is certainly correlated with the heterotrophic growth of C.kichenii.
E. As can be seen by measuring the change of the glucose content of the culture medium after 1 h of inoculation, the absorption speed of the chlorella keiskei with high chlorophyll content on the glucose is obviously faster than that of the chlorella keiskei with low chlorophyll content, and the addition of 60 mu g/LDCMU does not influence the absorption speed of the chlorella keiskei on the glucose. According to the literature, the glucose absorption of microalgae is through a homotropic proton channel, the process needs to consume ATP, and the supply of ATP is provided by a cyclic photosynthetic phosphorylation pathway which is mainly dominated by the photosynthetic system I. DCMU inhibits photosynthesis and calvin cycle by inhibiting electron transport on photosystem II, but does not affect the cyclic photosynthetic phosphorylation pathway that photosystem I dominates. Therefore, the inventor believes that the high chlorophyll content can promote the ATP production pathway of cyclic photosynthetic phosphorylation to improve the glucose absorption of the microalgae, and finally improve the mixotrophic growth speed of the microalgae.
Summarizing the above experimental conclusions, the inventors speculate that the mechanism of DCMU for improving the light tolerance of microalgae in the mixed culture growth state is as follows: 1.DCMU reduces the photo damage to microalgae by inhibiting the photosynthetic system II of microalgae, and reduces the degree of chlorophyll photo damage, so that microalgae can keep higher chlorophyll content under high light intensity; 2. the high chlorophyll content promotes a microalgae cyclic photosynthetic phosphorylation pathway which is dominant by a microalgae photosynthetic system I, and more ATP is provided to improve the absorption speed of the microalgae on glucose; 3. the photosynthesis promotion caused by the high chlorophyll content offsets the inhibition of DCMU to photosynthesis, so that the autotrophic speed of the microalgae is not influenced by the DCMU, and the higher glucose absorption and utilization speed improves the heterotrophic growth speed of the microalgae, so that the final heterotrophic growth speed of the microalgae is promoted.
Example 4:
(1) chlorella vulgaris (C. chevalieri) ((C. chevalieri))Parachlorella kessleri) FACHB-4 is a microalgae species, firstly, using BG11 culture solution to make enrichment culture, when the cell density of the Chlorella Kelvin reaches 1X 108～1×109When the seeds are cultured per mL, the Chlorella keiskei seed liquid is obtained. BG11 culture solution comprises the following components: NaNO3 (1.5 g/L)，K2HPO4 (40 mg/L)，MgSO4·7H2O (70 mg/L)，CaCl2·2H2O (40 mg/L), citric acid (6 mg/L), ferric ammonium citrate (6 mg/L), EDTA-Na2 (1 mg/L)，Na2CO3 (20 mg/L)，H3BO3 (2.86 mg/L)，MnCl2·4H2O (1.86 mg/L)，ZnSO4·7H2O (0.22 mg/L)，Na2MoO4·2H2O (0.39 mg/L)，CuSO4·5H2O (0.08 mg/L)，Co(NO3)2·6H2O (0.05 mg/L)，pH＝7.00。
(2) Preparing microalgae polyculture culture media with different glucose contents, wherein the culture medium is BG11 culture medium plus 2 g/L glucose and 2 g/L Tris alkali, and the pH value is 7.00.
(3) Inoculating the chlorella keiskei seed solution into microalgae mixed culture mediums with different glucose contents according to the volume ratio of 2%, adding DCMU with the final concentration of 60 mu g/L, and culturing the chlorella keiskei culture in an illumination incubator for 72 hours under the culture conditions: 30 ℃, 2000, 4000, 8000, 12000Lux illumination intensity, 16/8 hours illumination period. A control group (CK) was also set, with no DCMU added.
(4) The light tolerance and chlorophyll yield of the C.trivialis was measured by measuring the dry weight and chlorophyll content of the C.trivialis, and the results are shown in FIG. 4: after 24 hours of culture, the chlorophyll content of the chlorella keiskei decreases along with the increase of the illumination intensity; under the illumination intensity of 2000, 4000, 8000 and 12000Lux, the chlorophyll content of the Chlorella Kelly of the DCMU experimental group is 1.35, 2.02, 2.3 and 2.6 times of that of the blank control. After 72 hours of culture, the biological yield of the blank control is reduced after the illumination intensity is more than 4000 Lux, which shows that the growth of the Chlorella Kelvinella is inhibited by the over-strong light intensity, and the biomass of the experimental group added with DCMU shows a slightly rising trend. Under 2000, 4000, 8000 and 12000Lux illumination intensity, the biomass of the Chlorella Kelvin added in the DCMU experimental group is increased by 6.03%, 13.93%, 22.64% and 40.12% compared with the blank control. The result shows that the addition of DCMU can obviously improve the tolerance of the chlorella keiskei to high light intensity, and the higher the light intensity is, the more obvious the protection effect of DCMU on the chlorophyll of the chlorella keiskei and the more obvious the promotion effect on the mixed culture growth of the chlorella keiskei.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.

Claims (5)

  1. The application of DCMU in improving chlorophyll yield or strong light tolerance during microalgae polyculture fermentation is characterized by comprising the following steps: inoculating microalgae into a mixed culture medium containing DCMU for illumination culture;
    the microalgae is chlorella;
    the concentration of DCMU in the mixed culture medium is 60 mug/L;
    the mixed culture medium is a culture medium containing a carbon source;
    of said microalgaeThe cell density was 1X 108～1×109Per mL;
    the conditions of the light culture are as follows: the illumination intensity is 2000-12000 Lux.
  2. 2. The use of DCMU according to claim 1 for increasing chlorophyll production or strong light tolerance in microalgae polyculture fermentation, wherein:
    the inoculation amount of the microalgae is 1-3% of the volume percentage of the mixed culture medium.
  3. 3. The use of DCMU according to claim 1 for increasing chlorophyll production or strong light tolerance in microalgae polyculture fermentation, wherein: the carbon source is glucose.
  4. 4. The DCMU according to claim 1, wherein the DCMU is used for increasing chlorophyll production or strong light tolerance during microalgae polyculture fermentation, and the DCMU is characterized in that: the culture medium is BG11 culture medium.
  5. 5. The use of DCMU according to claim 1 for increasing chlorophyll production or strong light tolerance in microalgae polyculture fermentation, wherein:
    the conditions of the light culture are as follows: the temperature is 25-35 ℃.
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