Application of psoralidine in inhibition of algae growth and algae inhibition preparation

Abstract

The invention discloses an application of psoralidine in inhibition of algae growth and an algae inhibition preparation, and belongs to the technical field of water body environment ecological restoration, the inventor screens out a natural plant allelopathic substance-psoralidine through a large number of experiments, and the psoralidine is wide in raw material source, environmentally friendly, easy to degrade, free of secondary pollution and good in ecological safety. The psoralidine has the advantages that the psoralidine is green and environment-friendly, the effect of inhibiting the growth of algae is good, the broad-spectrum algae inhibition performance is realized, and the algae inhibited by the psoralidine comprise various algae such as microcystis aeruginosa, heteroflexus aeruginosa, skeletonema costatum and phaeocystis globosa; the method can be applied to inland lake algae bloom water bodies, can also be applied to ocean red tide algae bloom water bodies, can be applied to fresh water and ocean water bodies, and has a wide application prospect in the field of harmful algae treatment.

Description

Technical Field

[0001] The invention relates to the technical field of water environment ecological restoration, and in particular to the application of psoralen in inhibiting algae growth and an algae-inhibiting preparation. Background Art

[0002] Algae are important primary producers in aquatic ecosystems and are located at the beginning of the food chain. However, their excessive growth may cause a series of environmental problems, such as eutrophication of water bodies, algal blooms and accumulation of toxic substances, which seriously affect water quality and ecological balance and even threaten human health. In inland lakes or rivers, the most common harmful algae is Microcystis aeruginosa; in coastal waters of my country, harmful algae such as Heterosigma akashiwo, Skeletonema costatum, Phaeocystis globosa and Prorocentrum donghaiense are the dominant species that mainly cause outbreaks. In recent years, ecological problems caused by algae have become increasingly serious, especially in still or slowly flowing waters such as oceans, lakes and rivers, where algal blooms caused by excessive reproduction frequently occur.

[0003] Traditional algae control methods include physical, chemical and biological means. Although they can alleviate the problem of algae outbreaks to a certain extent, these methods have many limitations. For example, physical removal is costly and cumbersome, chemical agents may cause secondary pollution, and biological control effects are difficult to predict and may bring ecological risks such as the invasion of alien species. Therefore, finding new, safe and efficient algae suppression methods has become a key scientific issue that needs to be solved urgently.

[0004] In the prior art, researchers have also conducted some scientific research based on this. For example, the Chinese patent document with publication number CN105217801A discloses the application of environmental enzymes in preventing and controlling the harm of algae blooms in lakes, wherein the environmental enzymes are prepared by fermenting fruits and / or vegetables, as well as water and sugars in a closed container in proportion. The Chinese patent document with publication number CN102010072A discloses a method for inhibiting the growth of algae blooms by the allelopathic effect of aquatic plants, wherein a certain amount of water web algae is put into a eutrophic water body containing algae blooms, and the water web algae releases allelopathic substances under the induction of the algae blooms, and the allelopathic substances can effectively inhibit the growth of algae blooms. The Chinese patent document with publication number CN103395892A discloses the application of the dipeptide compound bacilysin in inhibiting the growth and reproduction of algae blooms, which has an inhibitory effect on Microcystis aeruginosa, but the above inventions are only for prevention and control of freshwater bodies.

[0005] As a potential green algae inhibitor, natural allelopathic substances have received extensive attention in recent years. For example, the Chinese patent document with publication number CN109264836A discloses the application of flavonoid allelopathic substances in Spartina alterniflora in inhibiting algae growth. Allelopathic effect refers to the process in which plants, microorganisms, etc. affect the growth of other organisms by releasing specific secondary metabolites. These natural allelopathic substances are widely available, low in toxicity, have little impact on the environment, and are biodegradable, so they have important research and application value in algae control. There are many kinds of plant secondary metabolites, including phenols, flavonoids, quinones and their derivatives, so it is promising to dig out new natural algae inhibitors from plant secondary metabolites. Summary of the invention

[0006] The present invention provides an application of psoralidin in inhibiting algae growth and an algae-inhibiting preparation. Psoralidin can be used to prevent and control blue algae blooms caused by Microcystis aeruginosa in freshwater, and red tides caused by Heterosigma akashiwo, Skeletonema costatum or Phaeocystis sphericalensis in the ocean, and can exert excellent effects at relatively low concentrations.

[0007] The specific technical solutions adopted are as follows:

[0008] The present invention provides an application of psoralen in inhibiting the growth of algae. The structural formula of psoralen is shown below:

[0009]

[0010] The algae inhibited by the psoralen include Microcystis aeruginosa, Heterosigma akashiwo, Skeletonema costatum or Phaeocystis sphericalensis; and the psoralen has a broad-spectrum algae inhibition effect.

[0011] The psoralen is derived from natural plants, including psoralen, Solidago canadensis, etc. It can be extracted and separated from natural plants according to the records of the prior art or directly purchased.

[0012] The inventors conducted a large number of experimental screenings and obtained a natural plant allelopathic substance - psoralen, which is environmentally friendly, easily degradable, will not produce secondary pollution, has good ecological safety, and has a good effect in inhibiting the growth of algae. Specifically, the functions of psoralen include but are not limited to inhibiting the growth of microalgae or killing microalgae. It can be applied to algal blooms in inland lakes and red tide algal blooms in the ocean. It can be used directly or made into an anti-algae preparation. It has broad application prospects in the field of harmful algae control.

[0013] The present invention also provides an algae-inhibiting preparation, which includes psoralen. When the algae-inhibiting preparation is used, the dosage of psoralen is greater than 0.125 mg / L, preferably greater than 0.25 mg / L, and most preferably 1.0 to 16 mg / L. Within the above most preferred range, the cost is low and a highly efficient algae-inhibiting effect can be achieved.

[0014] Specifically, the algae-inhibiting preparation includes but is not limited to powder, granule or liquid preparation, and its ingredients may also contain auxiliary agents or carriers, which play a protective role on psoralen, or are used to slowly release psoralen to enable it to exert a long-lasting algae-inhibiting effect.

[0015] The present invention also provides a method for controlling the growth of harmful algae in a water body, using the psoralen or the algae inhibitor; specifically, the psoralen or the algae inhibitor is added into the water body where harmful algae have erupted.

[0016] Water bodies with harmful algal blooms include blue-green algae blooms caused by Microcystis aeruginosa in freshwater, and red tides caused by Heterosigma akashiwo, Skeletonium costatum or Phaeocystis spherical in the ocean.

[0017] Preferably, when Microcystis aeruginosa is the dominant species in a water body where harmful algae blooms, the dosage of psoralen is maintained at greater than 0.34 mg / L.

[0018] Preferably, when Heterosigma akashiwo is the dominant species in a water body where harmful algae blooms, the dosage of psoralen is maintained at greater than 1.01 mg / L.

[0019] Preferably, when Skeletonema costatum is the dominant species in a water body where harmful algae bloom, the dosage of psoralen is maintained at greater than 0.41 mg / L.

[0020] Preferably, when Pseudocystis sphericalensis is the dominant species in a water body where harmful algae have bloomed, the dosage of psoralen is maintained at greater than 0.48 mg / L.

[0021] Experiments have shown that within the above preferred concentration range, the algae-inhibiting effect of the psoralen or the algae-inhibiting preparation is better.

[0022] Compared with the prior art, the present invention has the following beneficial effects:

[0023] (1) The present invention discovered that the natural allelopathic substance psoralen has excellent algae inhibition properties against harmful algae including Microcystis aeruginosa, Heterosigma akashiwo, Skeletonema costatum, and Phaeocystis sphericalensis.

[0024] (2) Psoralea corylifolia is derived from natural plants and can be extracted from Psoralea corylifolia and Solidago canadensis. It is environmentally friendly, easily degradable, does not cause secondary pollution, has good ecological safety, and is green and environmentally friendly.

[0025] (3) Psoralen has a highly effective inhibitory effect on a variety of algae. It can not only effectively inhibit the dominant cyanobacteria in freshwater algal blooms, but also effectively inhibit the growth of a variety of marine red tide algae. It has a broad spectrum and can be used to prevent and control harmful algae outbreaks in freshwater and marine waters. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 This is a growth curve of Microcystis aeruginosa inhibited by different concentrations of psoralen.

[0027] Figure 2 This is the fitting curve diagram of the effective inhibitory concentration of Psoralea corylifolia on Microcystis aeruginosa at 72 hours.

[0028] Figure 3 This is a growth curve of different concentrations of psoralen inhibiting the growth of Heterosigma akashiwo.

[0029] Figure 4 This is the fitting curve diagram of the effective inhibitory concentration of psoralen on Heterosigma akashiwo at 72 hours.

[0030] Figure 5 This is a growth curve of Skeletonema costatum inhibited by different concentrations of psoralen.

[0031] Figure 6 This is the fitting curve diagram of the effective inhibitory concentration of psoralen on Skeletonema costatum at 72 hours.

[0032] Figure 7 This is a growth curve of Psoralea corylifolia inhibited by different concentrations of Psoralea corylifolia.

[0033] Figure 8 This is the fitting curve of the effective inhibitory concentration of Psoralea corylifolia on Phaeocystis sphericalensis at 72 hours. DETAILED DESCRIPTION

[0034] In order to make the purpose, features and advantages of the present invention more obvious and easy to understand, the following is a detailed description through specific embodiments. In the following description, many specific details are set forth in order to fully understand the present invention. However, the present invention can be implemented in many other ways different from those described herein, and those skilled in the art can make similar improvements without violating the connotation of the present invention, so the present invention is not limited by the specific embodiments disclosed below. The technical features in each embodiment of the present invention can be combined accordingly without conflicting with each other.

[0035] The operating methods of the unrecorded specific conditions in the following examples are usually according to conventional conditions, or according to the conditions recommended by the manufacturer. The content not described in detail in this specification belongs to the prior art known to professionals in the field. The experimental materials adopted in the following examples, if not otherwise specified, can be purchased by conventional biochemical reagent companies. Psoralea corylifolia can be extracted and separated from natural plants (Psoralea corylifolia or Solidago canadensis) according to the records of the prior art or directly purchased.

[0036] In the following examples, the inhibitory effect of psoralen on Microcystis aeruginosa, Heterosigma akashiwo, Skeletonema costatum, and Phaeocystis sphericalensis was studied.

[0037] Example 1 Test of the anti-algae activity of psoralen against Microcystis aeruginosa

[0038] 16 mg of aladdin (>98%) was dissolved in 1 mL of dimethyl sulfoxide solvent to prepare a 16 mg / mL psoralen solution. Further, it was diluted to 0.125, 0.25, 1.0, 2.0, 4.0, 8.0, and 16.0 mg / mL using dimethyl sulfoxide as a working solution. Culture medium, logarithmic growth phase Microcystis aeruginosa, and 10 μL of working solution were added to a 6-well plate with a culture system of 10 mL so that the final concentrations of each experimental group were 0.125, 0.25, 1.0, 2.0, 4.0, 8.0, and 16.0 mg / L, and the experimental group with 10 μL of dimethyl sulfoxide was used as the control group.

[0039] The initial algae density was counted using a microscope and a blood cell counting plate, and the initial density was controlled to be 5×10 5 The tested algae species was Microcystis aeruginosa, the culture medium used was BG11 culture medium, the constant temperature and light incubator, the culture conditions were 25℃, the light intensity was 3000Lux, and the light-dark ratio was 12:12. After activation inoculation twice, the algae growth cycle was adjusted to the logarithmic phase, counted, and set aside.

[0040] After culturing the algae cells for 120 h, the cultured algae cells were sampled and counted regularly, and the inhibition rate was calculated. The calculation formula of the inhibition rate is as follows:

[0041]

[0042] Among them C sample is the algae density of the experimental group, C control is the algae density of the control group.

[0043] The experimental results are as follows Figure 1 As shown in the figure, it can be seen that psoralen can significantly inhibit the growth of Microcystis aeruginosa, and the inhibitory effect increases with the increase of concentration. Logistic regression analysis was further used to fit the inhibition rate, and the results are shown in the figure. Figure 2As shown, the results showed that the half-inhibitory concentration of psoralen to Microcystis aeruginosa was 0.34 mg / L (p<0.05), which was the optimal concentration for controlling freshwater algal blooms with Microcystis aeruginosa as the dominant species.

[0044] Example 2 Test of the anti-algae activity of psoralen against Heterosigma akashiwo

[0045] The algae species tested were Heterosigma akashiwo, and the culture medium used was f / 2 culture medium. The culture was placed in a constant temperature and light incubator, and the culture conditions were 25°C, light intensity of 3000 Lux, and light-dark ratio of 12:12. After activation inoculation twice, the algae growth cycle was adjusted to the logarithmic phase, counted, and set aside. The initial density of the experiment was 5×10 4 The other experimental steps and parameters were the same as those in Example 1.

[0046] The experimental results are as follows Figure 3 As shown in the figure, it can be seen that psoralen can significantly inhibit the growth of Heterosigma akashiwo, and the inhibitory effect increases with the increase of concentration. Logistic regression analysis was further used to fit the inhibition rate, and the results are shown in the figure. Figure 4 As shown, the results showed that the half-inhibitory concentration of psoralen to Heterosigma akashiwo was 1.01 mg / L (p<0.05), which is the optimal concentration for controlling marine red tides with Heterosigma akashiwo as the dominant species.

[0047] Example 3 Test of the anti-algae activity of psoralea corylifolia against Skeletonema costatum

[0048] The algae species was Skeletonema costatum, and the culture medium used was f / 2+Si culture medium. The algae were placed in a constant temperature and light incubator with the culture conditions of 25°C, 3000 Lux of light intensity, and 12:12 light-dark ratio. After activation inoculation twice, the algae growth cycle was adjusted to the logarithmic phase, counted, and set aside. The initial density of the experiment was 5×10 4 The other experimental steps and parameters were the same as those in Example 1.

[0049] The experimental results are as follows Figure 5 As shown in the figure, it can be seen that psoralen can significantly inhibit the growth of Skeletonema costatum, and the inhibitory effect increases with the increase of concentration. Logistic regression analysis was further used to fit the inhibition rate, and the results are shown in the figure. Figure 6 As shown, the results showed that the half-inhibitory concentration of psoralen to Skeletonema costatum was 0.41 mg / L (p<0.05), which is the optimal concentration for controlling marine red tides with Skeletonema costatum as the dominant species.

[0050] Example 4 Test of the anti-algae activity of psoralen against Psoralea sphericalensis

[0051] The spherical brown cyst was used as the test algae species, and the culture medium used was f / 2 culture medium. The culture was placed in a constant temperature and light incubator, and the culture conditions were 25°C, light intensity of 3000Lux, and light-dark ratio of 12:12. After activation inoculation twice, the algae growth cycle was adjusted to the logarithmic phase, counted, and set aside. The initial density of the experiment was 5×10 5 The other experimental steps and parameters were the same as those in Example 1.

[0052] The experimental results are as follows Figure 7 As shown in the figure, it can be seen that psoralen can significantly inhibit the growth of Psora sphericalensis, and the inhibitory effect increases with the increase of concentration. Logistic regression analysis was further used to fit the inhibition rate, and the results are shown in the figure. Figure 8 As shown, the results showed that the half-inhibitory concentration of psoralen to Skeletonema costatum was 0.48 mg / L (p<0.05), which is the optimal concentration for controlling marine red tides with Phaeocystis sphericalensis as the dominant species.

[0053] The embodiments described above provide a detailed description of the technical solutions of the present invention. It should be understood that the above are only specific embodiments of the present invention and are not intended to limit the present invention. Any modifications, supplements or similar substitutions made within the scope of the principles of the present invention should be included in the protection scope of the present invention.

Claims

1. The use of psoralen in inhibiting the growth of algae, characterized in that: The structural formula of Psoralea corylifolia is shown below: The algae inhibited by psoralen include Microcystis aeruginosa, Heterosigma akashiwo, Skeletonema costatum or Phaeocystis sphericalensis.

2. The use of psoralen in inhibiting algae growth according to claim 1, characterized in that: The psoralen is derived from natural plants, and the natural plants include psoralen or Solidago canadensis.

3. An algae-inhibiting preparation, characterized in that: The algae-inhibiting preparation includes psoralen.

4. The algae-inhibiting preparation according to claim 3, characterized in that: When the algae-inhibiting preparation is used, the dosage of psoralen is greater than 0.125 mg / L.

5. The algae-inhibiting preparation according to claim 3, characterized in that: The algae-inhibiting preparation is in the form of powder, granule or liquid preparation.

6. A method for controlling the growth of harmful algae in a water body, characterized in that: Psoralen or the algae-inhibiting preparation according to any one of claims 3 to 5 is used, and the psoralen or the algae-inhibiting preparation is put into a water body where harmful algae have bloomed.

7. The method for controlling the growth of harmful algae in water according to claim 6, characterized in that: When Microcystis aeruginosa is the dominant species in a water body where harmful algae blooms, the dosage of psoralen should be maintained at greater than 0.34 mg / L.

8. The method for controlling the growth of harmful algae in water according to claim 6, characterized in that: When harmful algae blooms and Heterosigma akashiwo is the dominant species in the water body, the dosage of psoralen should be kept greater than 1.01 mg / L.

9. The method for controlling the growth of harmful algae in water according to claim 6, characterized in that: When Skeletonegrum costatum is the dominant species in a water body where harmful algae bloom, the dosage of Psoralea corylifolia should be maintained at greater than 0.41 mg / L.

10. The method for controlling the growth of harmful algae in water according to claim 6, characterized in that: When harmful algae bloom in water bodies and Psoralea corylifolia is the dominant species, the dosage of Psoralea corylifolia should be maintained at greater than 0.48 mg / L.

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