CN113897309B

CN113897309B - Marine microbubble with broad-spectrum algicidal activity and application thereof

Info

Publication number: CN113897309B
Application number: CN202111107928.6A
Authority: CN
Inventors: 徐虹; 朱晓莹; 陈双双; 罗桂英; 谢婉馨
Original assignee: Xiamen University
Current assignee: Xiamen University
Priority date: 2021-09-22
Filing date: 2021-09-22
Publication date: 2023-08-18
Anticipated expiration: 2041-09-22
Also published as: CN113897309A

Abstract

The invention discloses a marine microbubble with broad-spectrum algae killing activity and application thereof, and a culture of the marine microbubble has obvious algae killing effect on various harmful red tide algae such as spherical brown algae, red tide heterowan algae, eastern sea protodinoflagellate and the like. The algae killing activity of the supernatant is kept stable under the environment of-80-100 ℃ and pH 4-12, has great potential for developing an algae killing bactericide, and has wide application prospect in the aspect of treating harmful algal bloom by a biological method.

Description

Marine microbubble with broad-spectrum algicidal activity and application thereof

Technical Field

The invention relates to a marine microbubble strain, in particular to a marine microbubble strain with broad-spectrum algicidal activity and application thereof. Belongs to the technical field of algae killing bacteria.

Background

Along with the rapid development of economy, the industrialization and the acceleration of the urban process, the water pollution and the eutrophication degree become serious, the algal bloom bursts are frequent, and the scale is also increased. Spherical brown algae (Phaeocystis globose) is a wide-temperature and wide-salt dinoflagellate widely distributed in a plurality of sea areas of the world, has high propagation speed, is a main algae species for forming harmful algal bloom, and is one of dominant algae species for frequently inducing algal bloom in south China sea areas [1-3]. The explosion can lead to death of a large amount of cultivated fishes, seriously endangers the marine environment and the sustainable development of the cultivation industry, and simultaneously the algae can generate hemolytic toxin to endanger human health [3-4]. Under the specific water environment, the spherical brown algae can also form a large colloidal capsule body to block the water intake of a cold water source of a nuclear power station, thereby threatening the safety of the nuclear power. Currently, methods for red tide management mainly include physical, chemical and biological methods. Compared with the physical method and the chemical method, which have the problems of high cost and secondary pollution, the biological method is widely focused by people because of the advantages of high efficiency, safety and the like, so that the screening, separation and identification of algae killing bacteria and the research for controlling the algae bloom become hot spots for controlling the algae bloom at present [5-6].

Algicidal bacteria are a generic term for bacteria that are capable of killing algae and lysing algae cells by contacting the algae or by itself producing a substance from metabolic activity. The present screening and isolation of algicidal bacteria has been carried out for various algal dominant species such as Microcystis aeruginosa, protoformazan east Asian algae, alexander taalexandrium, isodon akabane, skeletonema costatum, etc., and the group mainly includes Pseudoalteromonas (Pseudoalteromonas), vibrio (Vibrio) in the phylum gamma-proteobacteria, cellulophaga (Cellulophaga), flavobacterium (Flavobacterium) in the CFB group, and Bacillus (Bacillus) with low GC content [7-8]. But the research on algae killing microorganisms related to the spherical brown algae at home and abroad is relatively few, and the reports are Bacillus [9-14], streptomyces [15-17], hahella sp.KA22[18], and Microbacterium sp.CBA01[19].

In general, the currently reported algicides are essentially limited to algicidal activity against one or a few algae, with significant limitations in practical use.

Disclosure of Invention

The invention aims to overcome the defects of the prior art, and provides the marine microbubble strain with broad-spectrum algae killing activity and the application thereof, and the marine microbubble strain has good algae killing activity on various common algae and has wide application range.

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

1. marine microbubble bacteria (Microbulbifer maritimus) YX04 with broad-spectrum algicidal activity is deposited in the Guangdong province microorganism strain collection center for 2021, 3 months and 18 days, and deposited at the building 5 of No. 59 of the national institute of Mitrex 100 in Guangzhou city under the accession number GDMCC No.61568.

2. The marine microbubble strain is applied to the preparation of algicides or algicides.

Preferably, the algicidal species of the algicide include: diatom, xanthomonas, dinoflagellate, and Jinzheimers.

Further preferably, the algicidal species of the algicide include: pseudoalternaria, skeletonema wisii, skeletonema costatum, heterobay red tide algae, prototheca eastern sea, sargassum horneri, and phaeocystis globosa.

Still more preferably, the algicidal species of the algicide include: hachain alga of Wei's sea, skeletonema costatum, etc the anterior sulcus is strengthened and the coccygeum globosum is strengthened.

3. The application of the marine microbubble bacteria in the prevention and treatment of harmful algal bloom.

4. An algicide contains culture of above marine microbubble bacteria as effective component.

The invention has the beneficial effects that:

the invention spreads and streaks the marine microbubble strain YX04 flat plate, picks up single colony for culture, then switches on, and cultures for 48-72 hours to obtain strain culture solution. The culture solution is centrifuged to obtain supernatant, and the supernatant is filtered to obtain sterile supernatant, and the supernatant has a wide algae killing spectrum and has remarkable algae killing effect on various harmful red tide algae such as spherical brown algae, red tide iso-bay algae, prorocentrum donghaiense and the like. The algae killing activity of the supernatant is kept stable under the environment of-80-100 ℃ and pH 4-12. The strain fermentation broth supernatant provided by the invention can be used for efficiently killing various harmful algae, has great potential for developing an algicidal agent, and has wide application prospect in the aspect of treating harmful algal bloom by a biological method.

Drawings

FIG. 1 shows the growth curve of algicidal YX04 and the algicidal activity at different growth stages.

FIG. 2 is a morphological diagram of algicidal YX04. Wherein, (A) is the form of a flat colony, and (B) is a strain scanning electron microscope.

FIG. 3 is a phylogenetic tree constructed by strain YX04 based on the 16S rRNA gene sequence.

FIG. 4 is a graph showing algicidal activity of different components of the culture of strain YX04.

FIG. 5 shows algicidal activity of culture supernatants of different concentrations of bacteria.

FIG. 6 shows the structural changes of algal cells after treatment of the culture supernatant of 5% fungus YX04 with transmission electron microscope. Wherein a is a normal algal cell; b. c and d are algal cells treated with the culture supernatant for 24, 48 and 72 hours, respectively.

FIG. 7 shows the algicidal activity after treatment of the culture supernatant of the bacteria at different temperatures.

FIG. 8 shows the algicidal activity after culture supernatants of different pH-treated bacteria.

Preservation information

Classification naming: marine microbubble bacteria

Latin Wen Xueming: microbulbifer maritimus

Preservation unit name: guangdong province microorganism strain collection center (GDMCC)

Deposit unit address: guangzhou city first-violent middle road No. 100 college No. 59 building 5

Preservation date: 2021, 3, 18

Detailed Description

The invention is further illustrated in the following figures and examples, which are provided for the purpose of illustration only and are not intended to be limiting.

1. Isolation screening and identification of strains

(1) Taking a mangrove wetland soil sample in a national-level natural protection area of the campsis grandiflora, diluting with sterile seawater in a 10-time proportion gradient, uniformly mixing by shaking, respectively taking 0.2mL of the mixture to be coated on a Zobell2216E solid flat plate (5 g/L of bactopeptone, 1g/L of yeast extract, 0.1g/L of ferric phosphate, 15g/L of agar powder, and setting the volume of mature seawater to 1L, wherein agar powder is not added in a liquid culture medium, and culturing for 7d at 30 ℃ in a dark place;

(2) According to the differences of colony morphology, size, color and the like, different colonies are selected and inoculated into 20mL Zobell2216E liquid culture medium, placed in a shaking table (30 ℃ C., 150 rpm) and cultivated for 3d in a dark place;

(3) Re-streaking the culture on a Zobell2216E solid plate, and culturing for 3d at 30 ℃ in a dark place;

(4) Repeating the step (3) for a plurality of times until a pure culture is obtained;

(5) The spherical brown algae is provided by a algae seed library of an offshore marine environment science national emphasis laboratory of Xiamen university, and the culture medium used by the algae seed is f/2 culture medium (NaNO) ₃ 75mg/L,NaH ₂ PO ₄ .H ₂ O,Thiamine.HCl 0.1mg/L,Biotin 0.5μg/L,B ₁₂ 0.5μg/L,Na ₂ EDTA.2H ₂ O 4.36mg/L,FeCl ₃ .6H ₂ O 3.15mg/L,CuSO ₄ .5H ₂ O 0.01mg/L,ZnSO ₄ .7H ₂ O 0.022mg/L,CoCl ₂ .6H ₂ O 0.01mg/L,MnCl ₂ .4H ₂ O0.18mg/L,NaMoO ₄ .2H ₂ O0.006 mg/L, dissolved in aged seawater), the culture conditions were as follows: the illumination period is 12h/12h (illumination/darkness), and the illumination intensity is 50-100 mu mol of photons m ^-2 s ^-1 The temperature is 22-25 ℃. The spherical brown algae cultivated to the exponential growth phase are subpackaged in 50mL triangular flasks, 20mL of algae liquid is used for each flask, and the culture is adapted for 24 hours in an illumination incubator.

(6) Inoculating the purified single colony into 20mL Zobell2216E liquid culture medium, and placing into a shaking table at 30 ℃ for shake culture at 150rpm for 96 hours; the absorbance at 600nm was measured for each of the cultures of 0 hours, 2 hours, 4 hours, 6 hours, 8 hours, 10 hours, 12 hours, 15 hours, 18 hours, 20 hours, 22 hours, 26 hours, 30 hours, 36 hours, 48 hours, 54 hours, 74 hours, and fresh 2216E without inoculation was used as a blank baseline. And (5) drawing a growth curve. And respectively taking bacterial culture solutions for culturing for 18 hours, 24 hours, 42 hours, 48 hours, 66 hours, 72 hours and 90 hours, adding the bacterial culture solutions into the spherical brown algae liquid in the step (5) according to the proportion of 5% (v/v), and then placing the bacterial culture solutions in an illumination incubator for culturing. The control group was prepared by adding 5% (v/v) Zobell2216E medium to the algae solution, and 3 replicates were set in each group. Treating for 24h, measuring chlorophyll autofluorescence value with enzyme-labeled instrument or measuring cell number with cell counter, and calculating algae killing rate according to formula 1, thereby screening out strain with algae killing activity.

Algae killing rate (%) = (Nt-N0)/n0×100% (formula 1)

Wherein N0 is the chlorophyll autofluorescence value or the cell number of the algae cells for 0 h; nt is the chlorophyll autofluorescence value or the cell number of the algae cells corresponding to the treatment time.

(7) As shown in FIG. 1, the isolated strain YX04 has good algae killing effect, the strain grows for 24 hours to reach the stage growth period, and 43% of algae cells can be killed by treating the spherical brown algae with the bacterial liquid. The algae killing activity of the bacterial liquid is also enhanced along with the extension of the culture time, the algae killing rate of the spherical brown algae treated by the 48h bacterial culture liquid can reach 81 percent, and the algae killing rate of the bacterial liquid is stabilized at 92 to 95 percent when the culture time is prolonged to more than 66 h.

(8) The morphology of the flat plate and the scanning electron microscope of the strain YX04 is shown in figure 2, and the colony can form yellow smooth colony on the flat plate, which is bright yellow with metallic luster, and the scanning electron microscope observation shows that the bacterial cells are in a long rod shape, have a length of about 2-3 mu m and a width of about 0.2 mu m, have no flagella, and have smooth surfaces and no wrinkles. The strain YX04 is subjected to genome extraction and 16S rRNA gene amplification, and PCR products are connected through a T vector and then sent to Xiaomen platinum Rayleigh biotechnology sequencing. Sequences were removed with DNAstar and uploaded to the Korea EzBioCloud website (https:// www.ezbiocloud.net /) for alignment, and phylogenetic trees were constructed using the Neighbor-Joing method of MEGA7.0 software, the results of which are shown in FIG. 3. The strains YX04 and Microbulbifer maritimus TF-17 were found by comparison ^T The similarity of (2) was as high as 99.36%, and it was judged to be a genus of Microbacterium, and it was designated as Microbulbifer maritimus YX04. Its 16S rRNA sequence was uploaded to NCBI database under GenBank accession number MW425990.

2. Research on algae killing mode of algae killing strain

(1) The spherical brown algae cultivated to the exponential growth phase are subpackaged in 50mL triangular flasks, 20mL of algae liquid is used for each flask, and the culture is adapted for 24 hours in an illumination incubator.

(2) Single colonies of YX04 were picked and inoculated into 50mL of liquid 2216E medium, shake-cultured at 30 ℃ for 24 hours, then transferred into 150mL of fresh 2216E medium at a ratio (volume) of 1%, and placed in shake-culture at 30 ℃. Respectively taking bacterial culture solutions which are cultured for 24 hours, 48 hours and 72 hours, centrifuging at 8,000rpm for 10 minutes, separating thalli and supernatant, filtering the supernatant by a sterile 0.22 mu m filter membrane to obtain sterile supernatant, and re-suspending the thalli by f/2 sterile culture medium with the same volume as the bacterial culture solution;

(3) Sequentially treating spherical brown algae in the step (1) with bacterial culture supernatant, re-suspending bacterial cells and bacterial culture according to the volume ratio of 5%, measuring chlorophyll autofluorescence for 0h, 24h, 48h and 72h, setting three parallel experimental groups and calculating the algae killing rate. As a result, as shown in FIG. 4, 5% of the bacterial culture solution and 5% of the culture supernatant had strong algicidal effect, the algicidal rate for 24 hours was 64% and 81%, respectively, while the bacterial cells had no algicidal effect, which suggests that Microbulbifer maritimus YX04 was indirectly algicidal by secretion of algicidal active substances. Algae killing experiments were performed with 1%, 3%, 5% and 10% (v/v) of the supernatant of strain YX04 broth, respectively, with 2216E medium as a control, three replicates were used for each treatment group, and the algae killing activity was measured for 0h, 24h, 48h and 72 h. As shown in FIG. 5, as the concentration of the bacterial supernatant increased, the algae killing rate increased significantly, 1% by volume of the bacterial supernatant had only the effect of inhibiting the growth of algae without killing algae, while the 3% by volume of the bacterial supernatant treatment had an algae killing rate of 50% and the 5% by volume and 10% by volume of the bacterial supernatant treatment had an algae killing rate of 81% and 86% respectively, and as the treatment time increased, the algae killing rates of the 3% by volume, 5% by volume and 10% by volume treatment groups after 48 hours had algae killing rates of 55%,87% and 93%, and 72h algae killing rates of 63%,91% and 97%, respectively. Indicating that the algicidal activity of YX04 culture supernatant was concentration-dependent.

3. Effect of Strain YX04 culture supernatant on algal cell Structure

1) 5% by volume of the culture supernatant was treated with 15mL of algae solution for 24, 48 and 72 hours, centrifuged at 3000 to 4000rpm for 10min, and buffered with phosphate buffer pH 7.4,0.1M (8g NaCl,0.2g KCl,1.44g Na was weighed ₂ HPO ₄ ，0.24g KH ₂ PO ₄ Dissolving in 1L distilled water, adjusting pH to 7.4 with 1M HCl) 2 times, adding 500 μl mass concentrationGlutaraldehyde with a degree of 2.5%, and fixing for 2 hours at 4 ℃;

2) Centrifuging the fixed cells, removing supernatant, washing with phosphate buffer solution (pH 7.4,0.1M) for 3 times, each for 15min, and placing on a shaking table to accelerate washing;

3) Adding 1.2mL of melted low-melting-point agar with mass concentration of 2% into a 1.5mL centrifuge tube, inserting a 10 mu L gun head in the middle of the centrifuge tube, forming a small hole after the agar is cooled, suspending the algae cells in the small hole, sealing the small hole with agar, placing the agar on ice to solidify the agar, cutting the agar block packed with the algae cells into 1mm pieces with a blade ³ A plurality of small blocks of size;

4) Washing the pre-embedded sample with PBS for 3 times, 15min each time, and fixing in a post-fixing solution with the mass concentration of 1% osmium acid for 2h;

5) The phosphate buffer was washed three times for 15min each. The first two changes are carried out in a fume hood, and the waste washing liquid is put into a waste osmium acid collecting bottle;

6) First using H ₂ O: phosphate buffer = 1:1 (volume ratio) for 10min, then H ₂ O: phosphate buffer = 2:1 (volume ratio) for 5min, and finally H is used for ₂ O rinsing for one time;

7) Dehydrating for 20min by using 30% ethanol water solution by volume concentration, dehydrating for 20min by using 50% ethanol by volume concentration, and dyeing for overnight by uranium acetate;

8) Rinsing with 70% ethanol bucket for three times to remove uranium acetate, and dewatering with 90% ethanol and 100% ethanol for 3 times each for 15min;

9) Add 500. Mu.L ethanol: resin=1: 1 (volume ratio) for 1h, then adding 250 mu L of resin to permeate for 1h, adding resin for 2h, then adding resin for 1.5h, and standing overnight at 4 ℃; fresh resin is replaced, and the resin used in the experiment is LR White;

10 The resin is injected into the embedding holes of the embedding plate, and the resin occupies about half of the volume of the die. Slightly picking up a sample by using a toothpick, dipping the sample on dry and clean filter paper to absorb redundant raw resin, then placing the sample into an embedding hole, placing the sample in the embedding hole for 12 hours, straightening the sample under a dissecting mirror, slowly filling the sample with the resin, and placing the sample in a 60 ℃ oven for 2 days, wherein the liquid level is preferably equal to the upper edge of the mould;

11 Block and slice

The resin block was removed from the embedding mold and trimmed under a dissecting scope. The embedding block is first trimmed to a tetrahedron with a blade at an angle of about 45 deg.. The resin around the material is completely repaired as much as possible. The top surface is generally rectangular or trapezoidal in shape. Roughly cutting into 70nm thick ultrathin slices by using a Leica Mltracut R-type slicing machine, then clamping a copper net with a film by using tweezers, fishing out the slices, naturally airing the slices, and then placing the slices into a copper net box;

12 2% uranyl acetate-lead citrate double staining:

uranium dyeing: the plastic dyeing piece is bent and placed on a dyeing frame by adopting a small liquid drop dyeing method, a copper net is placed in a gap of the dyeing piece by using tweezers, the dyeing piece is carefully taken down, the flat plate shape is restored, and the copper net is clamped by the plastic gap. And (3) dropwise adding a newly configured water solution of 2% of uranyl acetate with mass concentration to one surface of the copper mesh covered by the film, and dyeing for 30 minutes in a dark place. Washed 5 times with double distilled water in a beaker, taken out and placed on filter paper. Cutting filter paper into strips, and sucking off excessive water;

lead dyeing: similar to uranium dyeing, a small droplet dyeing method is also adopted, and in order to prevent the sample from being polluted by lead carbonate generated by the reaction of carbon dioxide and dyeing, sodium hydroxide particles are required to be arranged around the droplets to adsorb carbon dioxide in the air. The dyeing is also protected from light, the cover is used for protection, in addition, the direct expiration of the sample being dyed is avoided as much as possible during the operation, and the cleaning operation is fast. After dyeing, washing and naturally airing, re-bending the dyeing piece on a dyeing frame, taking down a copper net and putting the copper net into a copper net box;

13 Using JEM-2100HC transmission electron microscope, photographed from low power to high power. As can be seen from fig. 6 a, the control coccoid cells are intact and nearly circular, the nuclei have obvious double membranes, and the nucleolus is darker; mitochondria are in a complete oval shape, and obvious ridges can be observed; the chloroplast lamellar structure is closely and orderly arranged. Adding YX04 sterile supernatant, treating for 24h (b in FIG. 6), wherein the cell membrane is incomplete, the cytoplasm is lost, the chloroplast lamellae are sparse and broken, and the mitochondrial ridge is not obvious; after 48 and 72 hours of treatment (c, d in FIG. 6), no apparent organelle structure was observed inside the cells, and the cells were completely vacuolated to form a large number of small vesicle structures.

4. Influence of temperature and pH on algicidal effect of YX04 bacteria culture supernatant

(2) The YX04 fermentation broth cultured for 72 hours was centrifuged at 10,000rpm for 10 minutes, and the supernatant was filtered through a sterile 0.22 μm filter membrane to obtain a sterile supernatant. The sterile supernatant obtained was divided into several 5mL portions. And (3) respectively placing each part of sterile supernatant at-80 ℃, at-20 ℃,4 ℃,20 ℃, 40 ℃, 60 ℃, 80 ℃ and 100 ℃ for 2 hours, taking out, placing at room temperature, adding the mixture into the algae liquid in the step (1) in a proportion (volume) of 5 percent after the temperature is stabilized at the room temperature, and measuring the algae killing rate after the treatment for 48 hours. The algae killing rate calculating method refers to formula 1. As shown in FIG. 7, the strain YX04 has strong algicidal effect at different temperatures (-80-100 ℃) and the algicidal effect of the supernatant is not significantly reduced even after being treated for 2 hours at a temperature above 100 ℃. This indicates that the algicidal activity of the YX04 algicidal substance has temperature stability.

(3) The YX04 fermentation broth was collected and cultured for 72 hours, centrifuged at 10,000rpm for 10 minutes, and the supernatant was filtered through a sterile 0.22 μm filter membrane to obtain a sterile supernatant. The obtained sterile supernatant was dispensed at a concentration of 5 mL/serving. Then, the pH value of each sterile supernatant was adjusted to 3, 4, 5, 6, 7, 8, 9, 10, 11 and 12 by using 6mol/L NaOH or HCl, respectively, and the mixture was allowed to stand for 2 hours, and then the pH of the different supernatants was adjusted to the initial pH of 8.5. The culture solution was added to the algae liquid cultured in step 1 at a ratio of 5%, and after 48 hours of treatment, the algae killing rate was measured. The algae killing rate calculating method refers to formula 1. As shown in FIG. 8, the strain YX04 has obvious algae killing effect under different pH values (3-12), and the algae killing effect of the bacterial culture supernatant is not affected by other pH values (4-12) except that the algae killing activity of the supernatant is reduced due to the treatment for 2 hours under the strong acid environment of the pH value 3. This indicates that the algicidal activity of the YX04 algicidal substance has acid-base stability.

5. Determination of the algicidal Spectrum of Strain YX04

(1) Testing15 kinds of algae (Cellulomycetes: phaeodactylum tricornutum (Phaeodactylum tricornutum), haematococcus pseudominifolia (Thalassiosira pseudonana), haematococcus wishlani (Thalassiosira weissflogii), plasmodium pterycinum (Amphibera alta), skeletonema costatum (Skeletonema costatum), chlorophyceae: dunaliella salina (Dunaliella salina), phaeophyllum stenocarum (Platymonas subcordiformis), chlorella vulgaris (Chlorella vulgaris), phaeophyllum qinghaosum (Platymonas helgolandica), chlorella viridis (Nannochloriopsis), xanthomonas: isodon aka (Heterosigma akashiwo), phaeophyceae: protorochaete eastern (Prorocentrum donghaiense), alexanthema minum (Alexandrium minutum), alexanthema robustum (Scrippsiella trochoidea), and Pleurospora sphaera (Phaeocystis globose)) were all supplied from the national laboratory of the offshore environmental sciences of the university of Euonymus, and the culture medium used as f/2 is as follows: the illumination period is 12/12h (illumination/darkness), and the illumination intensity is 50 mu mol photons m ^-2 s ^-1 The temperature was 25 ℃.

(2) After the algae cells are cultured to the logarithmic growth phase, sub-packaging the algae cells into 24-hole cell culture plates, wherein each hole is 2mL, each strain of algae is provided with 3 holes of a control group, the 3 holes of a treatment group are adapted to 24 hours in an illumination culture room;

(3) The bacteria YX04 sterile supernatant obtained by centrifugal filtration is added into an orifice plate at a concentration of 5% (volume), an equal amount of 2216E culture medium is added into a control group, chlorophyll autofluorescence values of algae cells are measured by an enzyme labeling instrument at regular intervals, and the algae killing activity is calculated according to a formula (1).

(4) As shown in Table 1, the strain YX04 mainly has good algae killing effect on various algae of diatoms such as Pseudostellaria, hachain Weissella, skeletonema costatum, isodon akabane, protopanaxaceae, phaeophyta and Phaeophyta, and has no algae killing effect on algae cells of Chlorophyta, indicating that the strain has broad-spectrum algae killing activity.

TABLE 1

While the foregoing description of the embodiments of the present invention has been presented with reference to the drawings, it is not intended to limit the scope of the invention, but rather, various modifications or variations can be made by those skilled in the art without the need of inventive effort on the basis of the technical solutions of the present invention.

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Claims

1. Marine microbubble strain with broad-spectrum algae killing activityMicrobulbifermaritimus) YX04, characterized in that it is deposited in the microorganism strain collection of Guangdong province for 2021, 3 months and 18 days, with a deposit address of Guangzhou Mitsui 100 th university, 59 th building 5, and a deposit number of GDMCC No.61568.

2. Use of marine microbubble bacteria according to claim 1 for the preparation of algicides or algicides, characterized in that the algicidal species of the algicides comprise: pseudoalternaria, skeletonema wisii, skeletonema costatum, heterobay red tide algae, prorocentrum donghaiense, sargassum horneri, and ascophyta globosum; the algae inhibiting type of the algae inhibiting agent comprises: pseudoalternaria, skeletonema wisii, skeletonema costatum, heterobay red tide algae, prototheca eastern sea, sargassum horneri, and phaeocystis globosa.

3. The use according to claim 2, wherein the algicidal species of algicide comprises: hachain alga of Wei's sea, skeletonema costatum, etc the anterior sulcus is strengthened and the coccygeum globosum is strengthened.

4. Use of marine microbubble bacteria according to claim 1 for controlling harmful algae, characterized in that the species of harmful algae comprise: pseudoalternaria, skeletonema wisii, skeletonema costatum, heterobay red tide algae, prototheca eastern sea, sargassum horneri, and phaeocystis globosa.

5. An algistat characterized in that the active ingredient is the culture of the marine microbubble according to claim 1.