CN116649355B

CN116649355B - Application of phenothiazine derivative in preparation of drugs or preparations for inhibiting blue algae growth

Info

Publication number: CN116649355B
Application number: CN202310656307.6A
Authority: CN
Inventors: 周治国; 王樱桥; 项大铖; 杨红
Original assignee: Shanghai Normal University
Current assignee: Shanghai Normal University
Priority date: 2023-06-05
Filing date: 2023-06-05
Publication date: 2025-04-25
Anticipated expiration: 2043-06-05
Also published as: CN116649355A

Abstract

The invention discloses application of a phenothiazine derivative in preparing a medicine or a preparation for inhibiting blue algae growth, wherein the phenothiazine derivative has a general structure shown as a formula (1), R ₁ is selected from F、Cl、Br、I、CF₃、CH₂F、CHF₂、C_mH_2m+1、OC_mH_2m+1,SC_mH_2m+1、N(C_mH_2m+1)₂,R₂, N (C _nH_2n+1)₂、OC_nH_2n+1、SC_nH_2n+1 and the like, m and N are integers from 0 to 12, the invention also provides a blue algae inhibitor which comprises one or more of the phenothiazine derivatives, the concentration of the phenothiazine derivative is 0.01-80 mug/mL, the phenothiazine derivative has remarkable inhibition effect on blue algae, the inhibition effect can reach more than 95%, the phenothiazine derivative has good selectivity on green algae under the same condition, and the phenothiazine derivative has remarkable inhibition effect on blue algae growth.

Description

Application of phenothiazine derivative in preparation of drugs or preparations for inhibiting blue algae growth

Technical Field

The invention belongs to the technical field of resources and environment, and particularly relates to application of a phenothiazine derivative in preparation of a medicine or a preparation for inhibiting blue algae growth.

Background

In recent years, harmful algae mass propagation (HABs) frequently occurs due to an increase in eutrophication of water bodies, and is exacerbated by global warming. Nutrient substances such as nitrogen, phosphorus and the like from domestic sewage, industrial and agricultural runoffs flow into a water body in a large quantity, so that algae cells overgrow, environmental factors (such as light intensity, temperature and pH value) can also influence harmful algal bloom, and the growth of algae can be further accelerated by temperature rise caused by climate change. The rapid growth of blue algae poses a great risk to the aquatic environment and to the health of aquatic organisms and humans.

Algae cells generally grow on the water surface, so that the light transmittance of the water body is reduced, the photosynthesis of aquatic plants is inhibited, the oxygen content of the water body is reduced, and the normal growth of the aquatic organisms is hindered. When harmful algal bloom explodes in raw water, the extremely high algae cell density greatly influences the efficiency of the water treatment process, and seriously influences the operation and water supply safety of a water works. In addition, the release of cyanobacteria anabolites into a body of water, such as malodorous substances and cyanobacteria toxins, particularly algal toxins, can have an irreversible adverse effect on aquatic organisms and humans. Microcystins are common, toxic and broad-spectrum algal toxins that, due to their stable and complex circulatory structure, cannot be ideally removed by waterworks. Lian et al have shown that microcystins can endanger the kidneys, liver and nervous system of the human body and, in severe cases, can lead to death if one is exposed to food or water contaminated with cyanobacterial toxins.

In the face of severe HABs pollution, the algae content of polluted water needs to be effectively reduced, and the existing algae inhibition method can be divided into three categories of physics, biology and chemistry according to the means, wherein the physical means is mainly to directly remove harmful algae in the water body by means of mechanical salvage, flocculation sedimentation, filtration and the like, and the flocculation sedimentation method is regarded as a promising strategy by using modified clay to flocculate and precipitate HAB cells. The biological means mainly inhibit excessive propagation of algae by means of biological manipulation, most of organisms such as fishes, shrimps and crabs eat algae in water, the introduction and the containment of the species can effectively prevent the formation of water bloom, and in addition, some heterotrophic bacteria have a certain inhibition effect on the growth of algae. The chemical means mainly comprises that algae cells in the water body are removed by adding chemical reagents with strong oxidizing property, chemical sensing effect or other functions into the water body containing a large amount of harmful algae or HABs, the chemical reagents have the advantages of quick effect, good selectivity and the like in algae removal, the chemical methods are more used for treating water bloom and red tide, and the common chemical algicides are divided into two main categories, namely inorganic algicides such as copper sulfate and organic algicides such as herbicides.

In summary, the physical algae inhibiting method has high cost and difficult popularization and use, the biological method has long-term effect, low cost and slow effect, improper use can affect the ecological system of the lake, and most of the currently used chemical algae killing agents have defects such as poor selectivity, easy resistance generation, low efficiency and large environmental pollution, and the invention is completed based on the fact.

Disclosure of Invention

The invention aims to provide application of a phenothiazine derivative in preparation of a medicine or a preparation for inhibiting blue algae growth, wherein the phenothiazine derivative has an inhibiting effect on blue algae, and solves the problems of poor selectivity and low efficiency of the existing chemical algicide.

In order to achieve the above purpose, the invention adopts the following technical scheme:

the invention provides application of a phenothiazine derivative in preparing a medicine or a preparation for inhibiting blue algae growth, wherein the phenothiazine derivative has a general structure shown in the following formula (1):

in formula (1), p is selected from integers from 0 to 12;

R ₁ is selected from F、Cl、Br、I、CF₃、CH₂F、CHF₂、C_mH_2m+1、OC_mH_2m+1、SC_mH_2m+1、N(C_mH_2m+1)₂,m and is selected from integers from 0-12;

R ₂ is selected from N (C _nH_2n+1)₂、OC_nH_2n+1、SC_nH_2n+1),

N is an integer from 0 to 12.

Preferably, in formula (1), p is selected from integers from 1 to 5.

Preferably, in formula (1), R ₁ is selected from one or more of Cl, CF ₃、CHF₂、SCH₃、OCH₃、OC₂H₅, or N (CH ₃)₂).

Preferably, in formula (1), R ₂ is selected from N (CH ₃)₂),

Preferably, the phenothiazine derivative is selected from any one of the following compounds:

preferably, the blue algae is synechocystis wc 6803.

Preferably, the medicament or preparation takes the phenothiazine derivative as an active ingredient, and the effective dose of the phenothiazine derivative is 0.01-80 mug/mL.

More preferably, the effective dose of the phenothiazine derivative is 5. Mu.g/mL.

The invention also provides a blue algae inhibitor which comprises one or more of the phenothiazine derivatives, and the concentration of the phenothiazine derivatives is 0.01-80 mug/mL, preferably 5 mug/mL.

Compared with the prior art, the phenothiazine derivative shown in the chemical formula (1) has a selective inhibition effect on blue algae, the inhibition effect can reach more than 95%, the phenothiazine derivative has good selectivity on green algae (taking chlorella pyrenoidosa as a model) under the same condition, particularly has no obvious killing effect on chlorella pyrenoidosa, and the phenothiazine derivative can be used for preparing medicines or preparations for inhibiting the growth of blue algae.

Drawings

FIG. 1 is a diagram showing the general structure of phenothiazine derivatives.

FIG. 2 shows (a) algicidal pictures of compound A1 at different concentrations and (b) inhibition rate in blue algae treated with compound A1 at different concentrations for 12h in the examples, wherein the illumination intensity is 100. Mu. Mol.m ^-2s^-1, the temperature is 27 ℃, and the initial cell density OD ₇₃₀ nm=1.5.

FIG. 3 shows (a) algicidal pictures of compound A2 at different concentrations and (b) inhibition rate in blue algae treated with compound A2 at different concentrations for 12h in the examples, wherein the illumination intensity is 100. Mu. Mol.m ^-2s^-1, the temperature is 27 ℃, and the initial cell density OD ₇₃₀ nm=1.5.

FIG. 4 shows (a) algicidal pictures of compound A3 at different concentrations and (b) inhibition rate in blue algae treated with compound A3 at different concentrations for 12h in the examples, wherein the illumination intensity is 100. Mu. Mol.m ^-2s^-1, the temperature is 27 ℃, and the initial cell density OD ₇₃₀ nm=1.5.

FIG. 5 shows (a) algicidal pictures of compound A4 at different concentrations and (b) inhibition rate in blue algae treated with compound A4 at different concentrations for 12h in the example, wherein the illumination intensity is 100 [ mu ] mol.m ^-2s^-1, the temperature is 27 ℃, and the initial cell density OD ₇₃₀ nm=1.5.

FIG. 6 shows (a) algicidal pictures of compound A5 at different concentrations and (b) inhibition rate in blue algae treated with compound A5 at different concentrations for 12h, wherein the illumination intensity is 100 [ mu ] mol.m ^-2s^-1, the temperature is 27-28 ℃, and the initial cell density OD ₇₃₀ nm=1.5.

FIG. 7 shows the inhibition rate of Chlorella pyrenoidosa 12h treated with compound A2 at different concentrations in the examples, wherein the light intensity is 100. Mu. Mol.m ^-2s^-1 and the temperature is 27-28deg.C.

FIG. 8 shows the inhibition rate of blue algae treated with compound A2 at different concentrations for 12h in the examples, wherein the illumination intensity is 100. Mu. Mol.m ^-2s^-1 and the temperature is 27-28 ℃.

FIG. 9 shows the inhibition of blue algae at an initial cell density OD ₇₃₀ nm=0.5 by compound A2 of 5. Mu.g/mL in the example, wherein the light intensity is 40. Mu. Mol.m ^-2s^-1 and the temperature is 27 ℃.

FIG. 10 shows the inhibition of blue algae at an initial cell density OD ₇₃₀ nm=1.0 by compound A2 of 5. Mu.g/mL in the example, wherein the light intensity is 40. Mu. Mol.m ^-2s^-1 and the temperature is 27 ℃.

FIG. 11 shows the inhibition of blue algae at an initial cell density OD ₇₃₀ nm=1.5 by compound A2 of 5. Mu.g/mL in the example, wherein the illumination intensity is 40. Mu. Mol.m ^-2s^-1 and the temperature is 27-28 ℃.

Detailed Description

In order to make the objects, technical solutions and advantages of the embodiments of the present invention more clear, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. It will be apparent that the described embodiments are some, but not all, embodiments of the invention. All other embodiments, which can be made by a person skilled in the art without creative efforts, based on the described embodiments of the present invention fall within the protection scope of the present invention.

The compounds A1 to A9 used in the examples below were derived from the references Chaudhari,A.,Pramanik,C.,Patil,P.,Vedpathak,H.,Bandishti,M.,Tripathy,N.K.,Gurjar,M.K.,J.Heterocycl.Chem.2022,59(6),1054.

Example 1

1. Algae cultivation

1.1 Method for culturing Chlorella pyrenoidosa

1.1.1 Preparation of mother liquor STOCKI:

1000mL of mother solution STOCKI (Hurner' S TRACE ELEMENTS) is prepared, wherein several components including NH ₄Cl(15.0g)、CaCl₂(1.5g)、MgSO₄ (3.2 g) and deionized water (with constant volume to 1000 mL) are added into a 1L beaker to be constant volume to 1000mL, and the mixture is stirred and mixed uniformly, then transferred into a glass bottle for sealing, sterilized at high temperature and high pressure and placed at normal temperature.

1.1.2 Preparation of mother liquor STOCKII:

STOCKII preparing the components of K ₂HPO₄(28.8g)、KH₂PO₄ (14.4 g) and deionized water (with constant volume to 100 mL), adding the components into a 500mL beaker, constant volume to 100mL, stirring, mixing uniformly, transferring into a glass bottle, sealing, sterilizing at high temperature and high pressure, and storing in a refrigerator with the temperature of 4 ℃.

1.1.3 Preparation of mother liquor STOCKIII:

STOCKIII preparation of ZnSO ₄·7H₂ O (15.0 g) dissolved in deionized water (50 mL), H ₃BO₃ (5.7 g) dissolved in deionized water (100 mL), mnCl ₂·4H₂ O (2.53 g) dissolved in deionized water (25 mL), co (NO ₃)₂·6H₂ O (0.9845 g) dissolved in deionized water (25 mL), cuSO ₄·5H₂ O (0.785 g) dissolved in deionized water (25 mL), na ₂MoO₄·2H₂ O (0.75375 g) dissolved in deionized water (25 mL), feSO ₄·7H₂ O (2.495 g) dissolved in deionized water (25 mL) and EDTA disodium salt (25 g) dissolved in deionized water (125 mL), STOCKIII preparation is sequentially dissolved in the above order, and then poured into a 500mL beaker in turn, the beaker is heated to boiling (heated uniformly) for multiple times, EDTA is poured into the beaker for multiple times to dissolve sufficiently, and the aqueous bath is carried out at 70 ℃ after slow pouring to prevent boiling, and the pH is adjusted to 6.7, the pH is greenish by KOH solution, the pH is adjusted to be 2.495, the solution is diluted to be in two weeks after the pH is adjusted to be 500 ℃ and finally filtered and stored in a refrigerator until the water is dark at 500 ℃ and the same time, the water is sterilized in a dark-black, and finally the solution is cooled in a refrigerator is sterilized.

Preparation of 1.1.4TAP culture medium:

After the three mother liquor formulations were completed, each of components STOCKI (125 mL), STOCKII (625. Mu.L), STOCKIII (5 mL), glacial acetic acid (5 mL), tris-base (12.1 g), deionized water (to volume of 5000 mL) was added to a 5L beaker. Adding secondary water to 5L, adjusting pH to 7.3, sterilizing at high temperature under high pressure, and storing at normal temperature. The mother liquor adding operation is carried out in an ultra clean bench.

1.1.5 Cultivation of Chlorella pyrenoidosa:

The study object is chlorella pyrenoidosa, all strains are grown in TAP culture medium with pH=7.3, the inoculation amount is 1%, and the culture is carried out in an illumination incubator with the temperature of 25 ℃ under continuous illumination with the illumination intensity of 40 mu E.m ^-2s^-1 for 36 hours, and after the culture is finished, the cell concentration is counted to be 2.0x10 ⁷-2.2×10⁷/mL by a cell counter.

1.2 Blue algae cultivation

1.2.1A5 preparation of mother solution:

a5 1L of formula, adding H₃BO₃(2.86g)、MnCI₂·4H₂O(1.81g)、ZnSO₄·7H₂O(0.222g)、Na₂MoO₄·2H₂O(0.39g)、CuSO₄·5H₂O(0.079g)、Co(NO₃)₂·6H₂O(0.0494g) components into a 1L beaker, fixing the volume to 1L, stirring, mixing uniformly, transferring into a glass bottle, sealing, sterilizing at high temperature and high pressure, and preserving at normal temperature.

Preparation of 1.2.3BGII culture medium:

BGII1L of formula is prepared by adding NaNO₃(1.5g)、K₂HPO₄·3H₂O(0.04g)、MgSO₄·7H₂O(0.075g)、CaCl₂·2H₂O(0.036g)、 citric acid (0.006 g), ferric citrate amine (0.006 g), EDTA-Na ₂(0.001g)、Na₂CO₃ (0.02 g), A5 (1 mL) and deionized water (1000 mL) into a 1L beaker, fixing the volume to 1L, adjusting pH to 8, stirring, mixing uniformly, transferring into a glass bottle, sealing, sterilizing at high temperature and high pressure, and preserving at normal temperature.

The cyanobacteria strain used in the following examples was Synechocystis pc 6803, all algal cells were cultured in BGII medium containing 5mM Tris-HCl at pH=8.0, the culture temperature was kept at about 30℃and the algal cells were cultured by continuous irradiation with an LED lamp having a light intensity of 40. Mu.E.m ^-2s^-1 and by introducing sterile air having a concentration of 2% (v/v) CO ₂ until OD ₇₃₀ nm=1.5.

2. Treatment of cyanobacteria cells

After blue algae or chlorella pyrenoidosa are cultured, 10mL of culture and phenothiazine derivatives with different concentrations are transferred into 60mL of hydrogen-producing glass tubes, the mouth of the hydrogen-producing tubes is sealed by a rubber plug and mixed by slight shaking, and the mixture is placed in an environment with the illumination intensity of 100 mu mol.m ^-2s^-1 and the temperature of 27-28 ℃ for a plurality of hours.

3. Inhibitory Effect of phenothiazine derivatives on Chlorella pyrenoidosa

Chlorella pyrenoidosa is cultured to different cell concentrations (5×10≡6/mL-2×10≡7/mL), 10mL of culture is transferred to each hydrogen-producing glass tube of 60mL, 0-800 μg of phenothiazine derivative is dissolved by 100 μl of DMSO (figure 1) respectively, 0-80 μg/mL of solution is obtained and transferred to the corresponding hydrogen-producing glass tube, the mouth of the hydrogen-producing tube is sealed by a rubber plug and is slightly shaken and mixed uniformly, and then the hydrogen-producing glass tube is placed in an environment with illumination intensity of 100 μmol.m ^-2s^-1 and temperature of 27-28 ℃ for 12 hours. After standing, extracting 5mL of algae liquid in each tube, centrifuging for 10min at 4000r/min, removing supernatant, vibrating on a vortex vibration instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuously vibrating on the vortex vibration instrument for 2 min, and then placing in a 4 ℃ refrigerator for standing for 12h. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating the inhibition rate of the chlorella pyrenoidosa.

4. Inhibition effect of phenothiazine derivative on blue algae

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10, 20 and 30mL of algae liquid are separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, the culture medium is transferred into a 60mL glass tube after shaking and suspending on a vortex oscillator, so as to obtain solutions with the cell density OD ₇₃₀ nm=0.5, 1.0 and 1.5, 0-800 mu g of phenothiazine derivative (figure 1) is respectively dissolved by 100 mu L of DMSO, the solutions with the concentration of 0-80 mu g/mL are transferred into corresponding hydrogen-producing glass tubes, the mouth of the hydrogen-producing tube is sealed by a rubber plug and mixed by slight shaking, and the solution is placed in an environment with the illumination intensity of 100 mu mol/m ^-2s^-1 and the temperature of 27-28 ℃ for 12 hours. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a 4 ℃ refrigerator for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating the inhibition rate of blue algae.

Example 2

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is removed, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the mixture is transferred into a 60mL glass tube, so as to obtain a solution with the cell density OD ₇₃₀ nm=0.5, and 0, 50, 100, 200, 400 and 800 mug of compound A1 are respectively dissolved by 100 mug of DMSOSolutions with concentrations of 0, 5, 10, 20, 40 and 80. Mu.g/mL were obtained and transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. Extracting 5mL of algae liquid in each tube after standing, centrifuging for 10min at 4000r/min, removing supernatant, vibrating on a vortex vibration instrument until no algae cells are attached to the wall of the centrifuge tube, adding 5mL of methanol, continuously vibrating on the vortex vibration instrument for 2 min, standing for 12h in a refrigerator at 4 ℃, centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating the inhibition rate of blue algae to be 0,20%,18%,30%,61% and 92% (figure 2).

Culturing Chlorella pyrenoidosa to cell concentration of 5×10ζ6/mL, transferring 10mL of culture to 60mL of hydrogen-producing glass tube, dissolving 0, 50, 100, 200, 400, 800 μg of compound A1 with 100 μl of DMSO to obtain 0, 5, 10, 20, 40, 80 μg/mL concentration solution, transferring into corresponding hydrogen-producing glass tube, sealing the mouth of hydrogen-producing tube with rubber plug, slightly shaking, mixing, standing at 27-28deg.C for 12 hr under light intensity of 100 μmol.m ^-2s^-1. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a 4 ℃ refrigerator for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,9%,8%,6%,6% and 3%.

Example 3

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the blue algae liquid is transferred into a 60mL glass tube, and the solution with the cell density OD ₇₃₀ nm=0.5 is obtained. Dissolving 0, 50, 100, 200, 400, 800 μg of Compound A2 with 100 μl of DMSO, respectivelyThe solution with the concentration of 0, 5, 10, 20, 40 and 80 mug/mL is obtained and transferred into a corresponding hydrogen producing glass tube, the mouth of the hydrogen producing tube is sealed by a rubber plug and is mixed by slight shaking, and then the mixture is placed in an environment with the illumination intensity of 100 mu mol-m ^-2s^-1 and the temperature of 27-28 ℃ for standing for 12 hours. After standing, extracting 5mL of algae liquid in each tube, centrifuging for 10min at 4000r/min, removing supernatant, vibrating on a vortex vibrating instrument until no algae cells are attached to the wall of the centrifuge tube, adding 5mL of methanol, continuously vibrating on the vortex vibrating instrument for 2min, and standing for 12h in a refrigerator at 4 ℃. Then centrifuging for 10min at 4000r/min, detecting absorbance value of supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating inhibition rate of blue algae to 0,23%,21%,30%,76% and 82% (figure 3).

Chlorella pyrenoidosa was cultured to a cell concentration of 5X 10≡6/mL, and 10mL of the culture was transferred to 60mL of each hydrogen-producing glass tube. Dissolving 0, 50, 100, 200, 400 and 800 mug of compound A2 with 100 mug of DMSO respectively to obtain 0,5, 10, 20, 40 and 80 mug/mL concentration solutions, transferring the solutions into corresponding hydrogen-producing glass tubes, sealing the mouths of the hydrogen-producing tubes with rubber plugs, slightly shaking and uniformly mixing the hydrogen-producing tubes, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,5%,2%,3%,3% and 1%.

Example 4

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the blue algae liquid is transferred into a 60mL glass tube, and the solution with the cell density OD ₇₃₀ nm=0.5 is obtained. Dissolving 0, 50, 100, 200, 400, 800 μg of Compound A3 with 100 μl of DMSO, respectivelyThe solution with the concentration of 0, 5, 10, 20, 40 and 80 mug/mL is obtained and transferred into a corresponding hydrogen producing glass tube, the mouth of the hydrogen producing tube is sealed by a rubber plug and is mixed by slight shaking, and then the mixture is placed in an environment with the illumination intensity of 100 mu mol.m ^-2s^-1 and the temperature of 27-28 ℃ for standing for 12 hours. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. Then centrifuging for 10min at 4000r/min, detecting absorbance value of supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating blue algae inhibition rate of 0,22%,28%,35%,70% and 80% (figure 4).

Culturing Chlorella pyrenoidosa to cell concentration of 5×10ζ6/mL, transferring 10mL of culture to 60mL of hydrogen-producing glass tube, dissolving 0, 50, 100, 200, 400, 800 μg of compound A3 with 100 μl of DMSO to obtain 0, 5, 10, 20, 40, 80 μg/mL concentration solution, transferring into corresponding hydrogen-producing glass tube, sealing the mouth of hydrogen-producing tube with rubber plug, slightly shaking, mixing, standing at 27-28deg.C for 12 hr under light intensity of 100 μmol.m ^-2s^-1. After standing, extracting 5mL of algae liquid in each tube, centrifuging for 10min at 4000r/min, removing supernatant, vibrating on a vortex vibrating instrument until no algae cells are attached to the wall of the centrifuge tube, adding 5mL of methanol, continuously vibrating on the vortex vibrating instrument for 2min, and standing for 12h in a refrigerator at 4 ℃. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,8%,5%,6%,7% and 4%.

Example 5

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the blue algae liquid is transferred into a 60mL glass tube, and the solution with the cell density OD ₇₃₀ nm=0.5 is obtained. Dissolving 0, 50, 100, 200, 400, 800 μg of Compound A4 with 100 μl of DMSO, respectivelyThe solution with the concentration of 0, 5, 10, 20, 40 and 80 mug/mL is obtained and transferred into a corresponding hydrogen producing glass tube, the mouth of the hydrogen producing tube is sealed by a rubber plug and is mixed by slight shaking, and then the mixture is placed in an environment with the illumination intensity of 100 mu mol-m ^-2s^-1 and the temperature of 27-28 ℃ for standing for 12 hours. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a 4 ℃ refrigerator for standing for 12 hours. Then centrifuging for 10min at 4000r/min, detecting absorbance value of supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating inhibition rate of blue algae to 0,18%,21%,30%,60% and 82% (figure 5).

Chlorella pyrenoidosa is cultured to a cell concentration of 5×10≡6/mL, 10mL of the culture is transferred to each hydrogen-producing glass tube of 60mL, 0, 50, 100, 200, 400 and 800 μg of compound A4 are dissolved by 100 μl of DMSO respectively to obtain solutions with concentrations of 0, 5, 10, 20, 40 and 80 μg/mL, the solutions are transferred to the corresponding hydrogen-producing glass tubes, the mouth of the hydrogen-producing tubes are sealed by rubber plugs and are slightly shaken and mixed uniformly, and the hydrogen-producing glass tubes are placed in an environment with illumination intensity of 100 μmol.m ^-2s^-1 and temperature of 27-28 ℃ for standing for 12 hours. After standing, extracting 5mL of algae liquid in each tube, centrifuging for 10min at 4000r/min, removing supernatant, vibrating on a vortex vibrating instrument until no algae cells are attached to the wall of the centrifuge tube, adding 5mL of methanol, continuously vibrating on the vortex vibrating instrument for 2 min, and standing for 12h in a refrigerator at 4 ℃. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,9%,11%,12%,8% and 7%.

Example 6

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is removed, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the mixture is transferred into a 60mL glass tube, so as to obtain a solution with the cell density OD ₇₃₀ nm=0.5, and 0, 50, 100, 200, 400 and 800 mug of compound A5 are respectively dissolved by 100 mug of DMSOThe solution with the concentration of 0, 5, 10, 20, 40 and 80 mug/mL is obtained and transferred into a corresponding hydrogen producing glass tube, the mouth of the hydrogen producing tube is sealed by a rubber plug and is mixed by slight shaking, and then the mixture is placed in an environment with the illumination intensity of 100 mug.m ^-2s^-1 and the temperature of 27 ℃ for standing for 12 hours. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. Then centrifuging for 10min at 4000r/min, detecting absorbance value of supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating inhibition rate of blue algae to 0,18%,21%,28%,78% and 83% (figure 6).

Chlorella pyrenoidosa is cultured to a cell concentration of 5×10≡6/mL, 10mL of the culture is transferred to each hydrogen-producing glass tube of 60mL, 0, 50, 100, 200, 400 and 800 μg of compound A6 are dissolved by 100 μl of DMSO respectively to obtain solutions with concentrations of 0, 5, 10, 20, 40 and 80 μg/mL, the solutions are transferred to the corresponding hydrogen-producing glass tubes, the mouth of the hydrogen-producing tubes are sealed by rubber plugs and are slightly shaken and mixed uniformly, and the hydrogen-producing glass tubes are placed in an environment with illumination intensity of 100 μmol.m ^-2s^-1 and temperature of 27-28 ℃ for standing for 12 hours. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,9%,11%,12%,8% and 7%.

Example 7

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is removed, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the mixture is transferred into a 60mL glass tube, so as to obtain a solution with the cell density OD ₇₃₀ nm=0.5, and 0, 50, 100, 200, 400 and 800 mug of compound A6 are respectively dissolved by 100 mug of DMSOSolutions with concentrations of 0, 5, 10, 20, 40 and 80. Mu.g/mL were obtained and transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. Extracting 5mL of algae liquid in each tube after standing, centrifuging for 10min at 4000r/min, removing supernatant, vibrating on a vortex vibration instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuously vibrating on the vortex vibration instrument for 2 min, placing into a4 ℃ refrigerator for standing for 12h, centrifuging for 10min at 4000r/min, detecting absorbance value of the supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating inhibition rate of 0,22%,23%,35%,62% and 85% on blue algae.

Culturing Chlorella pyrenoidosa to cell concentration of 5×10ζ6/mL, transferring 10mL of culture to 60mL of hydrogen-producing glass tube, dissolving 0, 50, 100, 200, 400, 800 μg of compound A6 with 100 μl of DMSO to obtain 0, 5, 10, 20, 40, 80 μg/mL concentration solution, transferring into corresponding hydrogen-producing glass tube, sealing the mouth of hydrogen-producing tube with rubber plug, slightly shaking, mixing, standing at 27-28deg.C for 12 hr under light intensity of 100 μmol.m ^-2s^-1. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a 4 ℃ refrigerator for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,10%,11%,10%,9% and 6%.

Example 8

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is removed, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the mixture is transferred into a 60mL glass tube, so as to obtain a solution with the cell density OD ₇₃₀ nm=0.5, and 0, 50, 100, 200, 400 and 800 mug of compound A7 are respectively dissolved by 100 mug of DMSOSolutions with concentrations of 0, 5, 10, 20, 40 and 80. Mu.g/mL were obtained and transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. Extracting 5mL of algae liquid in each tube after standing, centrifuging for 10min at 4000r/min, removing supernatant, vibrating on a vortex vibration instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuously vibrating on the vortex vibration instrument for 2 min, placing into a4 ℃ refrigerator for standing for 12h, centrifuging for 10min at 4000r/min, detecting absorbance value of the supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating inhibition rate of 0,18%,23%,45%,62% and 79% on blue algae.

Culturing Chlorella pyrenoidosa to cell concentration of 5×10ζ6/mL, transferring 10mL of culture to 60mL of hydrogen-producing glass tube, dissolving 0, 50, 100, 200, 400, 800 μg of compound A7 with 100 μl of DMSO to obtain 0, 5, 10, 20, 40, 80 μg/mL concentration solution, transferring into corresponding hydrogen-producing glass tube, sealing the mouth of hydrogen-producing tube with rubber plug, slightly shaking, mixing, standing at 27-28deg.C for 12 hr under light intensity of 100 μmol.m ^-2s^-1. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a 4 ℃ refrigerator for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,15%,13%,10%,9% and 6%.

Example 9

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is removed, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the mixture is transferred into a 60mL glass tube, so as to obtain a solution with the cell density OD ₇₃₀ nm=0.5, and 0, 50, 100, 200, 400 and 800 mug of compound A8 are respectively dissolved by 100 mug of DMSOSolutions with concentrations of 0, 5, 10, 20, 40 and 80. Mu.g/mL were obtained and transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. Extracting 5mL of algae liquid in each tube after standing, centrifuging for 10min at 4000r/min, removing supernatant, vibrating on a vortex vibration instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuously vibrating on the vortex vibration instrument for 2 min, placing into a4 ℃ refrigerator for standing for 12h, centrifuging for 10min at 4000r/min, detecting absorbance value of the supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating inhibition rate of 0,22%,23%,35%,62% and 85% on blue algae.

Culturing Chlorella pyrenoidosa to cell concentration of 5×10ζ6/mL, transferring 10mL of culture to 60mL of hydrogen-producing glass tube, dissolving 0, 50, 100, 200, 400, 800 μg of compound A8 with 100 μl of DMSO to obtain 0, 5, 10, 20, 40, 80 μg/mL concentration solution, transferring into corresponding hydrogen-producing glass tube, sealing the mouth of hydrogen-producing tube with rubber plug, slightly shaking, mixing, standing at 27-28deg.C for 12 hr under light intensity of 100 μmol.m ^-2s^-1. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a 4 ℃ refrigerator for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of the chlorella pyrenoidosa is 0,10%, 13%,9% and 7%.

Example 10

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is removed, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the mixture is transferred into a 60mL glass tube, so as to obtain a solution with the cell density OD ₇₃₀ nm=0.5, and 0, 50, 100, 200, 400 and 800 mug of compound A9 are respectively dissolved by 100 mug of DMSOSolutions with concentrations of 0, 5, 10, 20, 40 and 80. Mu.g/mL were obtained and transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. Extracting 5mL of algae liquid in each tube after standing, centrifuging for 10min at 4000r/min, removing supernatant, vibrating on a vortex vibration instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuously vibrating on the vortex vibration instrument for 2 min, placing into a4 ℃ refrigerator for standing for 12h, centrifuging for 10min at 4000r/min, detecting absorbance value of the supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating inhibition rate of 0,18%,23%,36%,62% and 85% on blue algae.

Culturing Chlorella pyrenoidosa to cell concentration of 5×10ζ6/mL, transferring 10mL of culture to 60mL of hydrogen-producing glass tube, dissolving 0, 50, 100, 200, 400, 800 μg of compound A9 with 100 μl of DMSO to obtain 0, 5, 10, 20, 40, 80 μg/mL concentration solution, transferring into corresponding hydrogen-producing glass tube, sealing the mouth of hydrogen-producing tube with rubber plug, slightly shaking, mixing, standing at 27-28deg.C for 12 hr under light intensity of 100 μmol.m ^-2s^-1. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a 4 ℃ refrigerator for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,8%,16%,13%,9% and 17%.

Example 11

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 20mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the blue algae liquid is transferred into a 60mL glass tube, and the solution with the cell density OD ₇₃₀ nm=1.0 is obtained. Dissolving 0, 50, 100, 200, 400 and 800 mug of compound A1 with 100 mug of DMSO respectively to obtain 0, 5, 10, 20, 40 and 80 mug/mL concentration solutions, transferring the solutions into corresponding hydrogen-producing glass tubes, sealing the mouths of the hydrogen-producing tubes with rubber plugs, slightly shaking and uniformly mixing the hydrogen-producing tubes, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a 4 ℃ refrigerator for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of blue algae is 0,18%,16%,32%,59% and 90%.

Chlorella pyrenoidosa was cultured to a cell concentration of 1X 10≡7/mL, and 10mL of the culture was transferred to 60mL of each hydrogen-producing glass tube. Dissolving 0, 50, 100, 200, 400 and 800 mug of compound A1 with 100 mug of DMSO respectively to obtain 0, 5, 10, 20, 40 and 80 mug/mL concentration solutions, transferring the solutions into corresponding hydrogen-producing glass tubes, sealing the mouths of the hydrogen-producing tubes with rubber plugs, slightly shaking and uniformly mixing the hydrogen-producing tubes, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,10%,7%,5%,6% and 4%.

Example 12

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 20mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, the culture medium is transferred into a 60mL glass tube after shaking and suspending on a vortex oscillator, so as to obtain the solution with the cell density OD ₇₃₀ nm=1.0, 0, 50, 100, 200, 400 and 800 mug of compound A2 are respectively dissolved by 100 mug of DMSO, 0, 5, 10, 20, 40 and 80 mug/mL of solution is obtained, the solution is transferred into a corresponding hydrogen-producing glass tube, the mouth of the hydrogen-producing tube is sealed by a rubber plug and is slightly shaken and mixed evenly, and the solution is placed in the environment with the illumination intensity of 100 mug.m ^-2s^-1 and the temperature of 27-28 ℃ for 12 hours. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of blue algae is calculated to be 0,20%,25%,34%,62% and 84%.

Chlorella pyrenoidosa was cultured to a cell concentration of 1X 10≡7/mL, and 10mL of the culture was transferred to 60mL of each hydrogen-producing glass tube. Dissolving 0, 50, 100, 200, 400 and 800 mug of compound A2 with 100 mug of DMSO respectively to obtain 0,5, 10, 20, 40 and 80 mug/mL concentration solutions, transferring the solutions into corresponding hydrogen-producing glass tubes, sealing the mouths of the hydrogen-producing tubes with rubber plugs, slightly shaking and uniformly mixing the hydrogen-producing tubes, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,6%,4%,5%,5% and 2%.

Example 13

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 20mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the blue algae liquid is transferred into a 60mL glass tube, and the solution with the cell density OD ₇₃₀ nm=1.0 is obtained. Dissolving 0, 50, 100, 200, 400 and 800 mug of compound A3 with 100 mug of DMSO respectively to obtain 0, 5, 10, 20, 40 and 80 mug/mL concentration solutions, transferring the solutions into corresponding hydrogen-producing glass tubes, sealing the mouths of the hydrogen-producing tubes with rubber plugs, slightly shaking and uniformly mixing the hydrogen-producing tubes, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a 4 ℃ refrigerator for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of blue algae is 0,23%,20%,33%,79% and 83%.

Chlorella pyrenoidosa was cultured to a cell concentration of 1X 10≡7/mL, and 10mL of the culture was transferred to 60mL of each hydrogen-producing glass tube. Dissolving 0, 50, 100, 200, 400 and 800 mug of compound A3 with 100 mug of DMSO respectively to obtain 0,5, 10, 20, 40 and 80 mug/mL concentration solutions, transferring the solutions into corresponding hydrogen-producing glass tubes, sealing the mouths of the hydrogen-producing tubes with rubber plugs, slightly shaking and uniformly mixing the hydrogen-producing tubes, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,9%,6%,5%,8% and 4%.

Example 14

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 20mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, the culture medium is transferred into a 60mL glass tube after shaking and suspending on a vortex oscillator, so as to obtain the solution with the cell density OD ₇₃₀ nm=1.0, 0, 50, 100, 200, 400 and 800 mug of compound A4 are respectively dissolved by 100 mug of DMSO, 0, 5, 10, 20, 40 and 80 mug/mL of solution is obtained, the solution is transferred into a corresponding hydrogen-producing glass tube, the mouth of the hydrogen-producing tube is sealed by a rubber plug and is slightly shaken and mixed evenly, and the solution is placed in the environment with the illumination intensity of 100 mug.m ^-2.s^-1 and the temperature of 27-28 ℃ for 12 hours. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of blue algae is 0,16%,23%,30%,78% and 81%.

Chlorella pyrenoidosa was cultured to a cell concentration of 5X 10≡6/mL, and 10mL of the culture was transferred to 60mL of each hydrogen-producing glass tube. Compound A4 was dissolved in 100. Mu.L of DMSO at 0, 50, 100, 200, 400, 800. Mu.g to give solutions at 0, 5, 10, 20, 40, 80. Mu.g/mL, and transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,13%,12%,10%,8% and 7%.

Example 15

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 20mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the blue algae liquid is transferred into a 60mL glass tube, and the solution with the cell density OD ₇₃₀ nm=1.0 is obtained. Dissolving 0, 50, 100, 200, 400 and 800 mug of compound A5 with 100 mug of DMSO respectively to obtain 0, 5, 10, 20, 40 and 80 mug/mL concentration solutions, transferring the solutions into corresponding hydrogen-producing glass tubes, sealing the mouths of the hydrogen-producing tubes with rubber plugs, slightly shaking and uniformly mixing the hydrogen-producing tubes, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of blue algae is calculated to be 0,20%,21%,28%,75% and 81%.

Chlorella pyrenoidosa was cultured to a cell concentration of 1X 10≡7/mL, and 10mL of the culture was transferred to 60mL of each hydrogen-producing glass tube. Compound A5 at 0, 50, 100, 200, 400, 800. Mu.g was dissolved in 100. Mu.L of DMSO, respectively, to give solutions at 0, 5, 10, 20, 40, 80. Mu.g/mL concentrations, which were transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,10%,11%,8%,8% and 7%.

Example 16

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, the culture medium is transferred into a 60mL glass tube after shaking and suspending on a vortex oscillator, so that a solution with the cell density OD ₇₃₀ nm=0.5 is obtained, and 0, 50, 100, 200, 400 and 800 mug of compound A6 are respectively dissolved by 100 mug of DMSO, so that solutions with the concentration of 0, 5, 10, 20, 40 and 80 mug/mL are obtained, and the solutions are transferred into corresponding hydrogen-producing glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. Extracting 5mL of algae liquid in each tube after standing, centrifuging for 10min at 4000r/min, removing supernatant, vibrating on a vortex vibration instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuously vibrating on the vortex vibration instrument for 2 min, placing into a 4 ℃ refrigerator for standing for 12h, centrifuging for 10min at 4000r/min, detecting absorbance value of the supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating inhibition rate of 0,22%,23%,35%,62% and 85% on blue algae.

Example 17

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, the culture medium is transferred into a 60mL glass tube after shaking and suspending on a vortex oscillator, so that a solution with the cell density OD ₇₃₀ nm=0.5 is obtained, and 0, 50, 100, 200, 400 and 800 mug of compound A7 are respectively dissolved by 100 mug of DMSO, so that solutions with the concentration of 0, 5, 10, 20, 40 and 80 mug/mL are obtained, and the solutions are transferred into corresponding hydrogen-producing glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. Extracting 5mL of algae liquid in each tube after standing, centrifuging for 10min at 4000r/min, removing supernatant, vibrating on a vortex vibration instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuously vibrating on the vortex vibration instrument for 2 min, placing into a 4 ℃ refrigerator for standing for 12h, centrifuging for 10min at 4000r/min, detecting absorbance value of the supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating inhibition rate of 0,18%,23%,45%,62% and 79% on blue algae.

Example 18

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, the culture medium is transferred into a 60mL glass tube after shaking and suspending on a vortex oscillator, so that a solution with the cell density OD ₇₃₀ nm=0.5 is obtained, and 0, 50, 100, 200, 400 and 800 mug of compound A8 are respectively dissolved by 100 mug of DMSO, so that solutions with the concentration of 0, 5, 10, 20, 40 and 80 mug/mL are obtained, and the solutions are transferred into corresponding hydrogen-producing glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. Extracting 5mL of algae liquid in each tube after standing, centrifuging for 10min at 4000r/min, removing supernatant, vibrating on a vortex vibration instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuously vibrating on the vortex vibration instrument for 2 min, placing into a 4 ℃ refrigerator for standing for 12h, centrifuging for 10min at 4000r/min, detecting absorbance value of the supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating inhibition rate of 0,22%,23%,35%,62% and 85% on blue algae.

Example 19

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, the culture medium is transferred into a 60mL glass tube after shaking and suspending on a vortex oscillator, so that a solution with the cell density OD ₇₃₀ nm=0.5 is obtained, and 0, 50, 100, 200, 400 and 800 mug of compound A9 are respectively dissolved by 100 mug of DMSO, so that solutions with the concentration of 0, 5, 10, 20, 40 and 80 mug/mL are obtained, and the solutions are transferred into corresponding hydrogen-producing glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. Extracting 5mL of algae liquid in each tube after standing, centrifuging for 10min at 4000r/min, removing supernatant, vibrating on a vortex vibration instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuously vibrating on the vortex vibration instrument for 2 min, placing into a 4 ℃ refrigerator for standing for 12h, centrifuging for 10min at 4000r/min, detecting absorbance value of the supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating inhibition rate of 0,18%,23%,36%,62% and 83% on blue algae.

Culturing Chlorella pyrenoidosa to cell concentration of 5×10ζ6/mL, transferring 10mL of culture to 60mL of hydrogen-producing glass tube, dissolving 0, 50, 100, 200, 400, 800 μg of compound A9 with 100 μl of DMSO to obtain 0, 5, 10, 20, 40, 80 μg/mL concentration solution, transferring into corresponding hydrogen-producing glass tube, sealing the mouth of hydrogen-producing tube with rubber plug, slightly shaking, mixing, standing at 27-28deg.C for 12 hr under light intensity of 100 μmol.m ^-2s^-1. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a 4 ℃ refrigerator for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,8%,16%,13%,9% and 16%.

Example 20

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 30mL of algae liquid is separated and centrifuged at the rotation speed of 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the solution is transferred into a 60mL glass tube, and the solution with the cell density OD ₇₃₀ nm=1.5 is obtained. Compound A1 was dissolved in 100. Mu.L of DMSO at 0, 50, 100, 200, 400, 800. Mu.g to give solutions at 0, 5, 10, 20, 40, 80. Mu.g/mL, and transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of blue algae is 0,21%,19%,31%,62% and 93%.

Chlorella pyrenoidosa was cultured to a cell concentration of 2X 10≡7/mL, and 10mL of the culture was transferred to 60mL of each hydrogen-producing glass tube. Compound A1 was dissolved in 100. Mu.L of DMSO at 0, 50, 100, 200, 400, 800. Mu.g to give solutions at 0,5, 10, 20, 40, 80. Mu.g/mL, and transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,8%,7%,5%,5% and 2%.

Example 21

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 30mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, the culture medium is transferred into a 60mL glass tube after shaking and suspending on a vortex oscillator, so that a solution with the cell density OD ₇₃₀ nm=1.5 is obtained, and 0, 50, 100, 200, 400 and 800 mug of compound A2 are respectively dissolved by 100 mug of DMSO, so that solutions with the concentration of 0, 5, 10, 20, 40 and 80 mug/mL are obtained, and transferred into corresponding hydrogen-producing glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of blue algae is 0,19%,25%,33%,62% and 83%.

Chlorella pyrenoidosa was cultured to a cell concentration of 2X 10≡7/mL, and 10mL of the culture was transferred to 60mL of each hydrogen-producing glass tube. Compound A2 was dissolved in 100. Mu.L of DMSO at 0, 50, 100, 200, 400, 800. Mu.g, to give solutions at 0,5, 10, 20, 40, 80. Mu.g/mL, and transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,6%,3%,5%,7% and 1%.

Example 22

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 30mL of algae liquid is separated and centrifuged at the rotation speed of 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the solution is transferred into a 60mL glass tube, and the solution with the cell density OD ₇₃₀ nm=1.5 is obtained. 0, 50, 100, 200, 400, 800. Mu.g of Compound A3 was dissolved in 100. Mu.L of DMSO, respectively, to give solutions at concentrations of 0, 5, 10, 20, 40, 80. Mu.g/mL, and transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of blue algae is 0,26%,21%,33%,79% and 81%.

Chlorella pyrenoidosa was cultured to a cell concentration of 2X 10≡7/mL, and 10mL of the culture was transferred to 60mL of each hydrogen-producing glass tube. 0, 50, 100, 200, 400, 800. Mu.g of Compound A3 was dissolved in 100. Mu.L of DMSO, respectively, to give solutions at concentrations of 0, 5, 10, 20, 40, 80. Mu.g/mL, and transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,7%,4%,5%,6% and 4%.

Example 23

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 30mL of algae liquid is separated and centrifuged at the rotation speed of 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the solution is transferred into a 60mL glass tube, and the solution with the cell density OD ₇₃₀ nm=1.5 is obtained. 0, 50, 100, 200, 400 and 800. Mu.g of phenothiazine compound A2 were dissolved in 100. Mu.L of DMSO, respectively, to give solutions at concentrations of 0, 5, 10, 20, 40 and 80. Mu.g/mL, and transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe by using a rubber plug, slightly shaking and uniformly mixing, and placing the pipe in an environment with the illumination intensity of 100 mu mol.m ^-2s^-1 and the temperature of 27-28 ℃. Standing for 12 hours. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. After 12h, centrifuging for 10min at 4000r/min, detecting absorbance value of supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating inhibition rate of blue algae to 0-2%, 3%,18%,39%,48% (figure 7).

Chlorella pyrenoidosa was cultured to a cell concentration of 5X 10≡7/mL, and 10mL of the culture was transferred to 60mL of each hydrogen-producing glass tube. 0, 50, 100, 200, 400 and 800. Mu.g of phenothiazine compound A2 were dissolved in 100. Mu.L of DMSO, respectively, to give solutions at concentrations of 0, 5, 10, 20, 40 and 80. Mu.g/mL, and transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe by using a rubber plug, slightly shaking and uniformly mixing, and placing the pipe in an environment with the illumination intensity of 100 mu mol.m ^-2s^-1 and the temperature of 27-28 ℃. Standing for 12 hours. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. After 12h, centrifuging for 10min at 4000r/min, detecting absorbance value of supernatant at 665nm, calculating to obtain chlorophyll a concentration, and calculating inhibition rate of blue algae to 0,20%,15%,50%,75% and 80% (figure 8).

Example 24

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 30mL of algae liquid is separated and centrifuged at the rotation speed of 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the solution is transferred into a 60mL glass tube, and the solution with the cell density OD ₇₃₀ nm=1.5 is obtained. Compound A5 at 0, 50, 100, 200, 400, 800. Mu.g was dissolved in 100. Mu.L of DMSO, respectively, to give solutions at 0, 5, 10, 20, 40, 80. Mu.g/mL concentrations, which were transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of blue algae is 0,19%,22%,29%,79% and 82%.

Chlorella pyrenoidosa was cultured to a cell concentration of 5X 10≡6/mL, and 10mL of the culture was transferred to 60mL of each hydrogen-producing glass tube. Compound A5 at 0, 50, 100, 200, 400, 800. Mu.g was dissolved in 100. Mu.L of DMSO, respectively, to give solutions at 0, 5, 10, 20, 40, 80. Mu.g/mL concentrations, which were transferred to corresponding hydrogen-generating glass tubes. Sealing the mouth of the hydrogen-producing pipe with a rubber plug, slightly shaking and uniformly mixing, and standing for 12 hours under the environment that the illumination intensity is 100 mu mol.m ^-2s^-1 and the temperature is 27-28 ℃. After standing, 5mL of algae solution in each tube is extracted from each tube, and the tubes are centrifuged for 10min at 4000 r/min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 5mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a refrigerator at 4 ℃ for standing for 12 hours. And centrifuging for 10min at 4000r/min, detecting the absorbance value of the supernatant at 665nm, and calculating to obtain chlorophyll a concentration, wherein the inhibition rate of chlorella pyrenoidosa is 0,8%,10%,11%,7% and 6%.

Example 25

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 10mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, the culture medium is transferred into a60 mL glass tube after shaking and suspending on a vortex oscillator, so as to obtain the solution with the cell density OD ₇₃₀ nm=0.5, 50 mug of compound A2 is respectively dissolved by 100 mug of DMSO, namely the concentration is 5 mug/mL, and the solution is transferred into the glass tube. Sealing the mouth of the hydrogen-producing pipe by using a rubber plug, slightly shaking and uniformly mixing, and then placing the pipe in an environment with the illumination intensity of 100 mu mol.m ^-2s^-1 and the temperature of 27-28 ℃. On days 0,1, 3, 5, 7, 3mL of algae liquid was photographed and extracted, and centrifuged at 4000r/min for 10min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 3mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a 4 ℃ refrigerator for standing for 12 hours. Then, the supernatant was centrifuged at 4000r/min for 10min, the absorbance at 665nm was measured, the chlorophyll a concentration was calculated, and the blue algae inhibition rate was calculated to be 0% on day 0, 13% on day 1, 48% on day 3, 48% on day 5, 37% on day 7, and the blue algae inhibition effect was found to be the best on the third day of compound A2 at a concentration of 5. Mu.g/mL (FIG. 9).

Example 26

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 20mL of algae liquid is separated and centrifuged at 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the blue algae liquid is transferred into a60 mL glass tube, and the solution with the cell density OD ₇₃₀ nm=1 is obtained. 50. Mu.g of Compound A2, i.e., 5. Mu.g/mL, was dissolved in 100. Mu.L of DMSO, respectively, and transferred into a glass tube. Sealing the mouth of the hydrogen-producing pipe by using a rubber plug, slightly shaking and uniformly mixing, and then placing the pipe in an environment with the illumination intensity of 100 mu mol.m ^-2s^-1 and the temperature of 27-28 ℃. On days 0,1, 3, 5, 7, 3mL of algae liquid was photographed and extracted, and centrifuged at 4000r/min for 10min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 3mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a 4 ℃ refrigerator for standing for 12 hours. Then, the supernatant was centrifuged at 4000r/min for 10min, the absorbance at 665nm was measured, the chlorophyll a concentration was calculated, and the blue algae inhibition rate was calculated to be 0% on day 0, 19% on day 1, 23% on day 3, 22% on day 5, and 22% on day 7, and the blue algae inhibition effect was found to be the best on the third day of compound A2 at a concentration of 5. Mu.g/mL (FIG. 10).

Example 27

After blue algae is cultured to the cell density OD ₇₃₀ nm=1.5, 30mL of algae liquid is separated and centrifuged at the rotation speed of 4000r/min, after supernatant is poured out, 30mL of fresh culture medium is added, and after shaking and suspending on a vortex oscillator, the solution is transferred into a 60mL glass tube, and the solution with the cell density OD ₇₃₀ nm=1.5 is obtained. 50. Mu.g of Compound A2, i.e., 5. Mu.g/mL, was dissolved in 100. Mu.L of DMSO, respectively, and transferred into a glass tube. Sealing the mouth of the hydrogen-producing pipe by using a rubber plug, slightly shaking and uniformly mixing, and then placing the pipe in an environment with the illumination intensity of 100 mu mol.m ^-2s^-1 and the temperature of 27-28 ℃. On days 0,1, 3, 5, 7, 3mL of algae liquid was photographed and extracted, and centrifuged at 4000r/min for 10min. After removing the supernatant, shaking on a vortex shaking instrument until no algae cells are attached to the wall of the centrifugal tube, adding 3mL of methanol, continuing shaking on the vortex shaking instrument for 2 minutes, and then placing in a 4 ℃ refrigerator for standing for 12 hours. Then, the supernatant was centrifuged at 4000r/min for 10min, the absorbance at 665nm was measured, the chlorophyll a concentration was calculated, and the blue algae inhibition rate was calculated to be 0% on day 0, 9% on day 1, 21% on day 3, 16% on day 5, and 13% on day 7, and the blue algae inhibition effect was found to be the best on the third day of compound A2 at a concentration of 5. Mu.g/mL (FIG. 11).

According to the embodiment, the phenothiazine derivative has a remarkable inhibition effect on blue algae, the inhibition effect can reach more than 95%, the phenothiazine derivative has no remarkable killing effect on chlorella pyrenoidosa under the same condition, and the phenothiazine derivative has good selectivity on green algae (taking chlorella pyrenoidosa as a model).

The previous description of the embodiments is provided to facilitate a person of ordinary skill in the art in order to make and use the present invention. It will be apparent to those skilled in the art that various modifications can be readily made to these embodiments and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above-described embodiments, and those skilled in the art, based on the present disclosure, should make improvements and modifications without departing from the scope of the present invention.

Claims

1. Use of a phenothiazine derivative selected from any one of the following compounds in the preparation of a medicament or formulation for inhibiting blue algae growth:

2. The use according to claim 1, wherein the cyanobacteria is synechocystis wc 6803.

3. The use according to claim 1, characterized in that the medicament or preparation contains the phenothiazine derivative as active ingredient and the effective dose is 0.01-80 μg/mL.

4. The use according to claim 1, characterized in that the effective dose of the phenothiazine derivative is 5 μg/mL.

5. A cyanobacterial inhibitor, characterized by comprising one or more of the phenothiazine derivatives described in claim 1, and the concentration of the phenothiazine derivatives is 0.01-80 μg/mL.

6. The cyanobacterial inhibitor according to claim 5, wherein the concentration of the phenothiazine derivative is 5 μg/mL.