CN105950500A - Algae-lysing aeromonas sp. and application thereof in controlling cyanobacterial blooms - Google Patents

Abstract

The invention discloses a strain of Aeromonas alginolyticus and its application in controlling cyanobacteria bloom. A strain of Aeromonas sp. (Aeromonas sp.) GLY‑2107 with remarkable algicidal activity was isolated from the water body of Taihu Lake, and its preservation number was CGMCC No.8979. Algal constituents 3‑benzyl‑piperazine‑2,5‑dione and 3‑methylindole, of which 3‑benzyl‑piperazine‑2,5‑dione was semi-lethal to Microcystis aeruginosa 9110 The dose LD50 is 4.72 μg/mL, and the semi-lethal dose LD50 of 3-methylindole to Microcystis aeruginosa 9110 is 1.10 μg/mL. It can be used in the research and development and production of new biological algicides, and finally applied to the control of cyanobacteria blooms in lakes.

Description

A strain of Aeromonas alginolyticus and its application in controlling cyanobacterial blooms

technical field

The invention relates to the field of environmental microbes, in particular to a strain of Aeromonas sp. GLY-2107 with algae-dissolving activity and its application in the control of cyanobacteria blooms.

Background technique

In recent years, Taihu Lake, Chaohu Lake, Dianchi Lake and other freshwater waters have been seriously affected by cyanobacteria blooms, and the cyanobacteria blooms caused by lake eutrophication have increasingly polluted lake water systems. Therefore, it is very necessary to explore effective ways to control the cyanobacterial biomass and inhibit the occurrence of cyanobacterial blooms.

The control technology of cyanobacteria bloom can be summarized as physical method, chemical method and biological method. Physical methods, such as mechanical algae removal, electromagnetic field algae removal, etc., can be used as auxiliary control measures for algae blooms, but the disadvantages are that they need a lot of manpower and material resources, and they can treat the symptoms but not the root cause, and the processing capacity is limited; chemical methods such as herbicides and other chemical algicides Algae can be directly killed, but the specificity of these chemicals is poor, and they are easy to accumulate in the food chain and cause secondary pollution.

The biological treatment method has attracted more and more attention because of its advantages of environmental protection and economy. Algae-lysing bacteria against cyanobacteria are a general term for bacteria that can directly (bacteria and algae cell contact) or indirectly (secretion of extracellular substances) inhibit and kill cyanobacteria. They are an important part of the freshwater ecological environment system. It plays an important role in reducing the biomass of cyanobacteria and maintaining ecological balance.

Therefore, those skilled in the art are committed to screening high-efficiency algicidal bacteria or isolating and enriching high-efficiency algicidal active substances produced by the metabolism of algicidal bacteria to develop microbial algicides for safe and efficient control of cyanobacteria blooms.

Contents of the invention

In view of the limited processing capacity of physical and chemical methods in the prior art, and the defects of easily causing secondary pollution, the technical problem to be solved in the present invention is to find efficient algae-dissolving bacteria and to separate and enrich the highly efficient algae-dissolving active substances produced by the metabolism of algae-dissolving bacteria , for safe and efficient control of cyanobacteria blooms.

In order to achieve the above object, the present invention provides the application of a strain of Aeromonas with alginolytic activity and its metabolites in the control of cyanobacteria blooms.

One aspect of the present invention provides a strain of Aeromonas (Aeromonas sp.) GLY-2107 with algicidal activity, which is Gram-negative, straight and short rod-shaped, polar single flagella, and the size is 0.6 μm to 1.2 μm. μm×2.0μm～2.5μm. After being cultured on the nutrient agar solid medium plate for 24 hours, the colony shape is round, the surface is smooth and flat, the edges are neat, white, and the colony size is 2mm to 3mm. After 16S rRNA gene sequence analysis and homology comparison, it was known that it had 99% homology with a certain Aeromonas strain in GenBank, so it was identified as Aeromonas bacteria and named Aeromonas GLY- 2107.

The bacterium was preserved in the General Microbiology Center of the China Committee for the Collection of Microbial Cultures, with the preservation number CGMCC No.8979, and the preservation date was March 28, 2014. Address of depository institution: Institute of Microbiology, Chinese Academy of Sciences, No. 1 Beichen West Road, Chaoyang District, Beijing, Zip Code: 100101, Tel: 86-10-64807355. The Aeromonas GLY-2107 can ferment to produce 3-benzyl-piperazine-2,5-dione and 3-methylindole.

Further, the present invention provides the application of the above-mentioned Aeromonas GLY-2107 in controlling cyanobacteria bloom.

Further, the present invention provides the application of the above-mentioned fermentation product of Aeromonas GLY-2107 in controlling cyanobacterial bloom, wherein the fermentation product can be fermentation broth, fermentation broth concentrate, fermentation broth crude extract or fermentation broth extract .

Another aspect of the present invention provides 3-benzyl-piperazine-2,5-dione and 3-methylindole, two active ingredients with algicidal activity in the fermentation product of Aeromonas GLY-2107. In the application of indole in controlling cyanobacteria bloom, the active ingredients in these two weeks have structures shown in structural formulas (I) and (II) respectively.

Another aspect of the present invention provides an algae-dissolving agent, which is characterized in that the algae-dissolving agent comprises the Aeromonas GLY-2107 described in claim 1.

Further, another aspect of the present invention provides an algicidal agent, characterized in that, the algicidal agent comprises the fermentation product of Aeromonas GLY-2107 according to claim 1, and the fermentation product is Fermentation broth, fermentation broth concentrate, fermentation broth crude extract or fermentation broth extract.

Yet another aspect of the present invention provides a kind of method of controlling cyanobacteria bloom, comprises the steps:

1), Aeromonas fermentum GLY-2107 strain;

2), extract the fermentation liquid;

3), the extract was evaporated to dryness to obtain a crude extract;

4) The crude extract is dissolved in water and used to control cyanobacteria blooms.

Preferably, in step 1), the fermentation condition is that Aeromonas GLY-2107 is inoculated in sterilized beef extract peptone medium with pH 7.0, and fermented at 28°C and 200rpm for 48 hours to obtain the Aeromonas GLY-2107 fermentation broth.

Preferably, in step 2), the extractant is ethyl acetate, and the volume ratio of ethyl acetate in the extraction system to the fermentation broth is 1:1. After mixing, put it in a shaker and vibrate for 24h, and the separated upper layer ethyl acetate The solution is the extract of Aeromonas GLY-2107 fermentation broth.

Preferably, the crude extract in step 3) is further purified to obtain effective algicidal substances for controlling cyanobacterial blooms. Wherein, further purification can adopt the method of column chromatography. Further preferably, the further purification is purified using HPLC technology, and the crude extract in step 3) is dissolved in water and filtered with a 0.22 μm pore size filter membrane, and the filtrate is passed through a Supersil ^TM C18-EP semi-preparative column of DIKMA Company, water and methanol The mobile phase was purified by HPLC to obtain effective algae-dissolving substances 2107-A and 2107-B.

The chemical structure of effective algicidal components in the metabolites of Aeromonas GLY-2107 is obtained through LC-MS, GC-MG and HMR analysis.

The quasi-molecular ion peak of the effective algae-dissolving component 2107-A is 205.0981m/z, the molecular weight of the sample is ^204.0899 , and the molecular formula is C ₁₁ H ₁₂ N ₂ O ₂ . ₆ ) δ8.15(1H,s,NH),7.89(1H,s,NH),7.35–7.11(5H,m,arom),4.06(1H,dd,J＝7.3,4.6Hz,H-6) ,3.37(1H,d,J=2.8Hz,H-3a),3.09(1H,dd,J=13.5,4.5Hz,H-7a),2.88(1H,dd,J=13.5,4.9Hz,H- 7b), 2.76 (1H, d, J=17.3Hz, H-3b); the quasi-molecular ion peak of the effective algae-dissolving component 2107-B is 132.0807m/z, the molecular weight of the sample is 131.0735, and the molecular formula is C ₉ H ₉ N , the result of H NMR spectrum is ¹ H NMR (600MHz, CDCl ₃ ) δ7.78 (1H, s, NH), 7.51 (1H, d, J=7.9Hz, H-4), 7.26 (1H, d, J ＝8.1Hz, H-7),7.14–7.09(1H,m,H-6),7.07–7.03(1H,m,H-5),6.88(1H,d,J＝1.0Hz,H-2) , 2.26 (3H, d, J = 1.1 Hz, Me).

The effective algicidal component 2107-A is the metabolite 3-benzyl-piperazine-2,5-dione of the Aeromonas GLY-2107, and the effective algicidal component 2107-B is the aeromonas 3-methylindole, a metabolite of Bacillus sp. GLY-2107.

The algalytic effect of the Aeromonas GLY-2107 of the present invention has a broad spectrum. The preparation method of the fermented liquid is simple and the preparation cycle is short. The fermentation broth of Aeromonas GLY-2107 has a good algalytic effect on Microcystis aeruginosa 9110, Synechococcus BN60, Chlorella B1, Oscillating algae BN35, etc. in Taihu Lake, and Microcystis aeruginosa is the cyanobacteria water A dominant cyanobacterium in Central China. Aeromonas GLY-2107 fermentation broth and ethyl acetate extract can be used to control cyanobacteria blooms or other microalgae, and the algae-dissolving effect is better. Among them, the 6-day algae-lysis rate of Aeromonas GLY-2107 fermentation broth to Microcystis aeruginosa 9110 reached 96.5±1.1%, the 6-day algae-lysis rate to Chlorella B1 reached 93.7±0.8%, and the 6-day algae-lysis rate to Microcystis aeruginosa The 6-day algae lysis rate of FACHB-979 reached 91.3±2.1%. Metabolites 3-benzyl-piperazine-2,5-dione and 3-methylindole of Aeromonas GLY-2107 have better lethality against dominant cyanobacteria Microcystis aeruginosa 9110 in cyanobacterial blooms in Taihu Lake 3-benzyl-piperazine-2,5-dione has a semi-lethal dose of LD50 of 4.72 μg/mL to Microcystis aeruginosa 9110; the semi-lethal dose of 3-methylindole to Microcystis aeruginosa 9110 LD50 is 1.10μg/mL, therefore, it can be used in the development and production of new algicides, and finally applied to the control of cyanobacteria blooms in lakes.

The effect of the Aeromonas GLY-2107 bacterial strain of the present invention and its metabolite 3-benzyl-piperazine-2,5-dione and 3-methylindole in the control of cyanobacteria blooms will be described below in conjunction with the accompanying drawings Further description is given to fully understand the purpose, features and effects of the present invention.

Description of drawings

Fig. 1 is the algalysis effect of alginolytic bacteria GLY-2107 to Microcystis aeruginosa 9110;

Fig. 2 is the algicidal effect of adding the ethyl acetate extract of the fermentation broth of alginolytic bacteria GLY-2107 to the Microcystis aeruginosa 9110 algae plate (the right side is the blank control);

Figure 3 is the algae-dissolving effect of the algae-dissolving substances 3-benzyl-piperazine-2,5-dione and 3-methylindole respectively added to the Microcystis aeruginosa 9110 plate (the far right is the blank control) ;

Figure 4 shows the algae-dissolving effects of the algae-dissolving substances 3-benzyl-piperazine-2,5-dione and 3-methylindole respectively added into the algae liquid on Microcystis aeruginosa 9110.

detailed description

The screening of embodiment 1 algae-lytic bacteria

Add 5mL of natural water samples collected from the Meiliang Bay waters of Taihu Lake to 95mL of algae liquid of Microcystis aeruginosa 9110 in the logarithmic phase. After the algae liquid is yellowed, it is serially diluted and coated with beef extract peptone medium agar plate, cultured at 28°C for 24 hours, and a plate with a moderate colony density is taken, and different strains are selected according to the different colony forms.

The screened strains were inoculated into 8 mL of beef extract peptone medium, cultured at 28°C and 220 rpm for 24 hours, and 1 mL of cultured bacterial liquid was added to 99 mL of algae liquid of Microcystis aeruginosa in logarithmic phase. In addition, 1 mL of sterilized beef extract peptone medium was also added to 99 mL of algae liquid as a control. All the algae fluids of the experimental group and the control group were cultured in the light incubator for 6 days to calculate their algicidal efficiency, and a strain of bacteria with a higher algicidal efficiency was selected for further research, and the strain code-named GLY-2107.

Algae liquid culture conditions: Microcystis aeruginosa 9110 was cultured with BG11 liquid medium, placed in a light incubator, 25°C, light intensity 40μmol photons m ^-2 s ^-1 , light-dark cycle ratio 12h:12h (light: dark ).

Calculation method of algae-dissolving efficiency: algae-dissolving efficiency (%)=(control group algae cell density-experimental group algae cell density)/control group algae cell density×100. The algae cell density of Microcystis aeruginosa was determined by hemocytometer.

Figure 1 is the curve of the concentration of Microcystis aeruginosa 9110 cells over time when algophilic bacteria GLY-2107 was added (algicidal bacteria GLY-2107 (2×10 ⁴ cells/mL) (▲) and blank control (■)).

Embodiment 2 Identification of Aeromonas GLY-2107 bacterial strain

GLY-2107, which has a strong algicidal effect, was identified by morphological observation, staining, physiological and biochemical reactions, and 16srRNA gene sequence analysis. μm～1.2μm×2.0μm～2.5μm. After being cultured on the nutrient agar solid medium plate for 24 hours, the colony shape is round, the surface is smooth and flat, the edges are neat, white, and the colony size is 2mm to 3mm. After 16S rRNA gene sequence analysis and homology comparison, it was known that it had 99% homology with a certain Aeromonas strain in GenBank, so it was identified as Aeromonas bacteria and named Aeromonas GLY- 2107. The strain has been preserved in the General Microbiology Center of the National Microbiological Culture Collection Management Committee, the preservation number is CGMCC No.8979, and the preservation date is March 28, 2014. Address of depository institution: Institute of Microbiology, Chinese Academy of Sciences, No. 1 Beichen West Road, Chaoyang District, Beijing, Zip Code: 100101, Tel: 86-10-64807355. The accession number of the 16srRNA gene of this strain in GenBank is KP717443.

The preparation method of embodiment 3 Aeromonas GLY-2107 fermented liquid and ethyl acetate extract thereof

Aeromonas GLY-2107 was inoculated in sterilized beef extract peptone medium according to 1% inoculation amount, and cultured on a shaker at 220 rpm at 28° C. for 48 hours to obtain Aeromonas GLY-2107 fermentation broth. Add ethyl acetate into the fermentation broth at a ratio of 1:1, put it into a shaker and vibrate for 24 hours, and separate the upper layer solution, that is, the ethyl acetate extract. The ethyl acetate was evaporated to dryness, dissolved in water, and filtered through a microporous membrane with a pore size of 0.22 μm for further purification of metabolites.

Embodiment 4 Aeromonas GLY-2107 is to the algicidal effect of different cyanobacteria and eukaryotic algae

Add 1 mL of Aeromonas GLY-2107 bacteria liquid of 8 cultured (culture conditions: beef extract peptone medium, 28°C, 220 rpm for 24 hours) to the algal liquid of 8 strains of cyanobacteria and eukaryotic algae (Table 1) (99mL, all algae liquids were cultured to the logarithmic phase), and 1 mL of sterile beef extract peptone medium was added to 8 kinds of algae liquids (99mL, all algae liquids were cultured to the logarithmic phase) as a control (1:99 ), after cultivating in the light incubator for 6 days, measure the chlorophyll concentrations of the algae fluids of all experimental groups and control groups respectively, and calculate the algae-dissolving efficiency. Three parallel samples were measured for each sample, expressed in the form of mean ± standard deviation.

Three of the algal species used in the experiment were purchased from the Algae Species Bank of Wuhan Institute of Hydrobiology, and the other five were screened from the water body of Taihu Lake. The experimental results show that the fermentation broth of Aeromonas GLY-2107 has a good algicidal effect on most prokaryotic algae species, but does not show a good algicidal effect on Chromococcus FACHB-191 (Table 1). The results indicated that Aeromonas GLY-2107 has a broad-spectrum algicidal ability and is a very potential algicidal bacterium.

Table 1. Algicidal effect of Aeromonas GLY-2107 on 8 strains of algae

Note: The algae strains marked with an asterisk (*) in the table were screened from the water body of Meiliang Bay in Wuxi, Taihu Lake, and the rest of the algae strains were purchased from the China Freshwater Algae Species Bank.

The algicidal effect of Aeromonas GLY-2107 on the dominant cyanobacteria Microcystis aeruginosa 9110 in the Taihu Lake cyanobacteria bloom is shown in Figure 1. Figure 1 shows that after adding Aeromonas GLY-2107, the concentration of Microcystis aeruginosa 9110 cells decreased with time, and the algae lysis rate was 96.5% on the sixth day. In the blank control, the concentration of Microcystis aeruginosa 9110 cells increased with time. The initial concentration of Aeromonas GLY-2107 was 2×10 ⁴ cells/mL.

Example 5 Research on algae-dissolving mode of GLY-2107 strain

The Aeromonas GLY-2107 fermentation broth was made into ethyl acetate extract according to the method described in step 3, evaporated to dryness with a centrifugal dryer, dissolved in sterile water and set aside. Finally, the sterilized beef extract peptone medium was also centrifugally dried and concentrated as a control. The extract prepared according to the above method and the concentrated beef extract peptone control were respectively added to a disc (5 mm in diameter) and placed on a BG11 solid plate made of Microcystis aeruginosa. Put the algae plate into the light incubator and cultivate it for 2 days, then observe the formation of the algae inhibitory circle around the disc to judge its algicidal effect.

The preparation method of the Microcystis aeruginosa agar plate: Pour an appropriate amount (about 20 mL) of BG11 agar medium (agar ratio 1.5%) onto each plate, and wait for it to solidify for later use. Centrifuge 300mL of the cultured Microcystis aeruginosa liquid at 4000g for 20min, discard the supernatant, collect the algal cell pellet and add it to 100mL of sterilized BG11 soft agar solid medium (1% agar, placed in a water bath at 53°C to prevent solidified), shake well and pour into the upper layer of prepared BG11 solid plate (approximately 20mL per plate), put it into the light incubator after solidification and cultivate it for later use.

The experimental results are shown in Figure 2, showing that the fermentation liquid extract of Aeromonas GLY-2107 has a good algicidal effect, which indicates that the strain has the ability to secrete extracellular algicidal substances for algicidal performance.

Example 6 Extraction and Purification of Algicidal Active Components in Aeromonas GLY-2107 Fermentation Product

The algicidal metabolites of the algicidal bacteria GLY-2107 were purified by HPLC technology to obtain two purified substances 2107-A and 2107-B with algicidal functions. The specific steps are as follows:

The extract of Aeromonas GLY-2107 fermentation broth was preliminarily purified through Supersil ^TM C18-EP semi-preparative column of DIKMA Company, using water and methanol as the mobile phase HPLC technical means to obtain algae-dissolving active ingredients. Then it was further purified by HPLC with DIKMA Supersil ^TM C18-EP analytical column and water and methanol as the mobile phase to obtain two effective algae-dissolving components 2107-A and 2107-B for structural identification.

Example 7 Identification of the Chemical Structure of the Algicidal Active Components in the Metabolites of Aeromonas GLY-2107

Using LC-MS﹑GC-MG and HMR analysis (hydrogen nuclear magnetic resonance spectrum analysis) the chemical structure of the metabolites with algicidal function.

The purified sample 2107-A was analyzed by LC-MS, and the quasi-molecular ion peak of sample 2107-A was 205.0981 m/z, the sample molecular weight was 204.0899, and the molecular formula was C ₁₁ H ₁₂ N ₂ O ₂ .

The purified sample 2107-A was analyzed by GC-MS, and it was similar to the structure of 3-benzyl-piperazine-2,5-dione in the GC-MS database, and the similarity index was >900.

Sample 2107A was subjected to proton nuclear magnetic resonance analysis, and the following results were obtained:

¹ H NMR (600MHz, DMSO-d ₆ ) δ8.15 (1H, s, NH), 7.89 (1H, s, NH), 7.35–7.11 (5H, m, aroma), 4.06 (1H, dd, J= 7.3,4.6Hz,H-6),3.37(1H,d,J＝2.8Hz,H-3a),3.09(1H,dd,J＝13.5,4.5Hz,H-7a),2.88(1H,dd, J=13.5, 4.9Hz, H-7b), 2.76 (1H,d, J=17.3Hz, H-3b).

Analyzing the LC-MS, GC-MS and NMR results of sample 2107-A, it can be determined that the metabolite 2107-A with algicidal effect is 3-benzyl-piperazine-2,5-dione, and its chemical The structure is shown in the following formula.

The purified sample 2107-B was analyzed by LC-MS, and the quasi-molecular ion peak of sample 2107-B was 132.0807m/z, the molecular weight of the sample was 131.0735, and the molecular formula was C ₉ H ₉ N.

The purified sample 2107-B was analyzed by GC-MS, and the structure of 2107-B was similar to that of 3-methylindole in the GC-MS database, and the similarity index was >900.

Sample 2107-B was subjected to proton nuclear magnetic resonance analysis, and the following results were obtained:

¹ H NMR (600MHz, CDCl ₃ ) δ7.78 (1H, s, NH), 7.51 (1H, d, J = 7.9Hz, H-4), 7.26 (1H, d, J = 8.1Hz, H-7 ),7.14–7.09(1H,m,H-6),7.07–7.03(1H,m,H-5),6.88(1H,d,J＝1.0Hz,H-2),2.26(3H,d, J = 1.1 Hz, Me).

Analyzing the LC-MS, GC-MS and NMR results of sample 2107-B, it can be confirmed that the metabolite 2107-B with algicidal effect is 3-methylindole, and its chemical structure is shown in the following formula.

Example 8 Algae-dissolving Efficiency of Aeromonas GLY-2107 Algae-dissolving Metabolites 3-Benzyl-piperazine-2,5-dione and 3-Methylindole

Using the above-mentioned method in the study of the algae-dissolving mode of the GLY-2107 strain, it was found through experiments that 3-benzyl-piperazine-2,5-dione and 3-methylindole were in the BG11 solid produced by Microcystis aeruginosa On the flat plate, obvious algae inhibition circles can be formed, while the blank control does not form algae inhibition circles (Fig. 3).

3-Benzyl-piperazine-2,5-dione and 3-methylindole were respectively added to the dominant cyanobacterium Microcystis aeruginosa 9110 in Taihu Lake to make 3-benzyl-piperazine-2,5-dione The concentration of ketone and 3-methylindole form a series respectively: 50μg/mL﹑40μg/mL﹑30μg/mL﹑20μg/mL﹑10μg/mL, 5.0μg/mL, 4.0μg/mL, 3.0μg/mL, 2.5 μg/mL, 2.0 μg/mL, 1.5 μg/mL, 1.0 μg/mL, 0.8 μg/mL, 0.6 μg/mL, 0.4 μg/mL, 0.2 μg/mL, and 0.1 μg/mL. Microcystis aeruginosa 9110 added with 3-benzyl-piperazine-2,5-dione or 3-methylindole was placed in a light incubator for 24 hours and then the algae dissolution rate was determined. The experimental results showed that the semi-lethal dose LD50 of 3-benzyl-piperazine-2,5-dione to Microcystis aeruginosa 9110 was 4.72 μg/mL; The lethal dose LD50 was 1.10 μg/mL (Figure 4). The results indicated that the metabolites 3-benzyl-piperazine-2,5-dione and 3-methylindole of Aeromonas GLY-2107 were very effective substances for controlling cyanobacteria.

Figure 4 shows the change curve of the survival rate of Microcystis aeruginosa 9110 when adding different concentrations of algicidal substance 3-benzyl-piperazine-2,5-dione and different concentrations of algicidal substance 3-methylindole.

The algalytic effect of the Aeromonas GLY-2107 of the present invention has a broad spectrum. The preparation method of the fermented liquid is simple and the preparation period is short. The fermentation broth of Aeromonas GLY-2107 has a good algalytic effect on Microcystis aeruginosa 9110, Synechococcus BN60, Chlorella B1, Oscillating algae BN35, etc. in Taihu Lake, and Microcystis aeruginosa is the cyanobacteria water A dominant cyanobacterium in Central China. Aeromonas GLY-2107 fermentation broth and ethyl acetate extract can be used to control cyanobacteria blooms or other microalgae, and the algae-dissolving effect is better. Among them, the 6-day algalysis rate of Aeromonas GLY-2107 fermentation broth to Microcystis aeruginosa 9110 reached 96.5±1.1%. Metabolites 3-benzyl-piperazine-2,5-dione and 3-methylindole of Aeromonas GLY-2107 have better lethality to the dominant cyanobacterium Microcystis aeruginosa 9110 in the cyanobacterial bloom of Taihu Lake It can be used in the research and development and production of new algicides, and finally applied to the control of cyanobacteria blooms in lakes.

The preferred specific embodiments of the present invention have been described in detail above. It should be understood that those skilled in the art can make many modifications and changes according to the concept of the present invention without creative effort. Therefore, all technical solutions that can be obtained by those skilled in the art based on the concept of the present invention through logical analysis, reasoning or limited experiments on the basis of the prior art shall be within the scope of protection defined by the claims.

Claims (10)

1. a strain has Aeromonas (Aeromonas sp.) GLY-2107 of alga-lysing activity, is preserved in Chinese micro-life Thing culture presevation administration committee's common micro-organisms center, preserving number CGMCC No.8979, preservation date is On March 28th, 2014, described Aeromonas GLY-2107 can ferment and produce 3-benzyl-piperazin-2,5-diketone With 3-methylindole.

2. the Aeromonas GLY-2107 as claimed in claim 1 application in controlling blue-green alga bloom.

3. the tunning of Aeromonas GLY-2107 as claimed in claim 1 is in controlling blue-green alga bloom Application.

What Aeromonas GLY-2107 the most according to claim 1 produced has shown in structural formula I and II The compound 3-benzyl-piperazin-2,5-diketone of structure and the application in controlling blue-green alga bloom of the 3-methylindole.

5. a molten phycomycete agent, it is characterised in that comprise the gas list described in claim 1 in described molten phycomycete agent Born of the same parents bacterium GLY-2107.

6. a molten algae medicament, it is characterised in that comprise the gas list described in claim 1 in described molten algae medicament The tunning of born of the same parents bacterium GLY-2107, described tunning is that fermentation liquid, concentrate of fermentation liquid, fermentation liquid slightly carry Thing or fermentation broth extract.

7. the method controlling blue-green alga bloom, comprises the steps:

1), fermentation Aeromonas GLY-2107 described in claim 1；

2), extractive fermentation liquid；

3), extract is evaporated, it is thus achieved that crude extract；

4) it is used for after, crude extract is water-soluble controlling blue-green alga bloom.

8. method as claimed in claim 7, wherein step 1) in, fermentation condition is, Aeromonas GLY-2107 It is inoculated in the sterilizing beef-protein medium of pH 7.0,28 DEG C, 220rpm environmental condition bottom fermentation 48h, Obtain described Aeromonas GLY-2107 fermentation liquid.

9. method as claimed in claim 7, wherein step 2) in, extractant is ethyl acetate, acetic acid second Ester is 1:1 with the volume ratio of fermentation liquid.

10. method as claimed in claim 7, wherein, step 3) in crude extract by purifying further Obtain effective algicidal substances, be used for controlling blue-green alga bloom.