Use of alpha-lindenic acid in inhibiting metamorphosis development of cyanea nozakii

Abstract

The application belongs to the technical field of jellyfish prevention and control, and discloses application of alpha-Lindenic acid in inhibiting metamorphosis development process of Aurelia coerulea, and establishes an experimental model for intervention of alpha-Lindenic acid in metamorphosis development of A.coerulea. When alpha-Lindenic acid is mixed with 5-methoxy-2-methylindole to induce polyp metamorphosis development of Aurelia coerulea, the mixed solution with a final concentration of 1 μM obviously inhibits the process of metamorphosis development of Aurelia coerulea. In addition, alpha-Lindenic acid reduces the number of discoid body release, and provides a new theoretical basis for preventing jellyfish outbreak.

Description

Technical Field

[0001] This invention relates to the field of jellyfish control technology, and more specifically, to the application of α-Lindenic acid in inhibiting the metamorphosis process of moon jellyfish. Background Technology

[0002] The moon jellyfish (Aurelia coerulea) is an ancient organism that lives near the coast. Moon jellyfish exhibit a generational cycle, with clear morphological transitions observed between polyps and medulla. Typically, the trumpet-shaped polyp has a basal disc and tentacles. When conditions for fission reproduction are met, it reproduces by fission and gradually develops into a fissile body. Once the fissile body grows, it detaches from the parent body, flips over in the sea to become an independent disc-shaped body, and begins a free-floating life, eventually developing into a medulla larva. This process of the polyp transforming into a disc-shaped body is called "metamorphosis." Under suitable conditions for metamorphosis, a single polyp can evolve into several or even a dozen medulla larvae, causing a multiplicative increase in jellyfish populations. Consequently, moon jellyfish outbreaks occur frequently in many sea areas worldwide, placing enormous pressure on marine ecosystems and negatively impacting human activities and the economy.

[0003] The transverse slit stage is the most important part of the entire metamorphosis process, especially the early transverse slit, which is the key to metamorphosis. Inhibiting the occurrence of early transverse slits, shortening the transverse slit cycle, and reducing the number of segmental discs are important strategies for controlling jellyfish populations and reducing jellyfish outbreaks.

[0004] Currently, there is limited research on the prevention and control of moon jellyfish, and effective prevention and control measures are lacking. Summary of the Invention

[0005] To overcome the aforementioned problems in existing technologies, the application of α-Lindenic acid in inhibiting the metamorphosis process of moon jellyfish is first proposed.

[0006] The objective of this invention is achieved through the following technical solution:

[0007] Application of α-Lindenic acid in inhibiting the metamorphosis process of moon jellyfish.

[0008] Preferably, α-Lindenic acid has the following effects:

[0009] (1) Delay the process of hydromorphic development;

[0010] (2) Reduce the number of disc-shaped objects released;

[0011] (3) Delay the time for discoid body release to reach its peak;

[0012] (4) Delay the time when early transverse fractures begin to appear.

[0013] Preferably, in the above application, α-Lindenic acid and the polyp of the moon jellyfish are incubated together, wherein the concentration of α-Lindenic acid is 1 μM.

[0014] This invention also provides the application of α-Lindenic acid in the preparation of formulations that inhibit the metamorphosis process of moon jellyfish.

[0015] Preferably, in the application of the above-mentioned preparation, α-Lindenic acid has the following effects:

[0016] (1) Delay the process of hydromorphic development;

[0017] (2) Reduce the number of disc-shaped objects released;

[0018] (3) Delay the time for discoid body release to reach its peak;

[0019] (4) Delay the time when early transverse fractures begin to appear.

[0020] As one specific implementation method, the above-mentioned application of the present invention under laboratory conditions includes the following steps:

[0021] Step 1: Prepare a 50 μM 5-methoxy-2-methylindole solution using artificial seawater to induce metamorphosis in A. coerulea;

[0022] Step 2: Prepare a 12-well plate, set up a control group and an experimental group, and randomly select 30 uniformly sized and vigorous hydra bodies for each group. Set up three replicates for each group, that is, place 10 hydra bodies in each well.

[0023] Step 3: Weigh 1 mg of α-Lindenic acid and add 359.16 μL of DMSO to obtain a 10 mM stock solution;

[0024] Step 4: Take 10 μL of the mother liquor and add 9.99 mL of seawater mixture (seawater containing 50 μmol / L 5-methoxy-2-methylindole) to obtain 10 μM dilution 1;

[0025] Step 5: Take 10 mL of dilution solution 1 and 90 mL of seawater mixture to obtain 1 μM α-Lindenic acid treatment solution, which has a volume of 100 mL. Dispense the solution into new 50 mL centrifuge tubes for later use.

[0026] Step 6: Add 2 mL of 5-methoxy-2-methylindole induction solution to each well in the control group, and add an equal volume of 1 μM α-Lindenic acid treatment solution to the experimental group. Incubate in a constant temperature incubator at 20 °C.

[0027] Step 7: Starting from the time the treatment solution is added, observe and record the time and number of different transverse splitting stages (early and late stages), as well as the time and number of released discoid bodies and completed transverse splitting individuals.

[0028] Preferably, the salinity of the artificial seawater in step one is 28‰–32‰. Too high or too low salinity will affect the health of the polyps.

[0029] Compared with the prior art, the present invention has the following beneficial effects:

[0030] This invention established an experimental model of α-lindenic acid intervention in the metamorphosis of *Aurelia coerulea*. When α-lindenic acid was mixed with 5-methoxy-2-methylindole to induce polyp metamorphosis in moon jellyfish (*Aurelia coerulea*), a final concentration of 1 μM significantly inhibited the metamorphosis process. Furthermore, α-lindenic acid reduced the number of discoid bodies released, providing a new theoretical basis for preventing jellyfish blooms. Attached Figure Description

[0031] Figure 1 The results showed the appearance of early, intermediate, and late transverse splits in the hydroids of the control group and the 1 μM α-Lindenic acid intervention group, as well as the time and proportion of transverse split completion.

[0032] Figure 2 This study compares the number of discoids released daily by A. coerulea hydroids between the control and intervention groups during the experimental period. Detailed Implementation

[0033] The technical solution of the present invention will be clearly and completely described below with reference to the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of the present invention.

[0034] I. Experimental Materials

[0035] 5-Methoxy-2-methylindole: Chemically induced metamorphosis of A. coerulea using a 50 μmol / L induction solution prepared with artificial seawater at 28‰-32‰.

[0036] α-Lindenic acid: Prepare a mixed treatment solution with a final concentration of 1 μM according to the instructions for use.

[0037] The apparatus for culturing hydra was a constant temperature incubator provided by Shanghai Lichen Instrument Technology Co., Ltd.

[0038] II. Experimental Methods

[0039] 1. Preparation before the experiment

[0040] Two weeks prior to the experiment, healthy and uniformly sized polyps were randomly selected from the hydroid tank and inoculated into a new 12-well plate. The experiment consisted of two groups: a control group and an inhibitor intervention group (1 μM). Each group had three replicates (n=3), with 10 polyps per well, for a total of 60 polyps. The health of the polyps was observed one week after inoculation. Feeding began after complete attachment, occurring every two days, with fresh seawater replaced 2-3 hours after feeding. The polyps were fasted for three days prior to the experiment to maintain peak vitality.

[0041] 2. Preparation of experimental reagents

[0042] 50 μM 5-methoxy-2-methylindole solution: Weigh 12.2 mg of 5-methoxy-2-methylindole and place it in a 50 mL beaker. Add 30 mL of prepared artificial seawater and stir with a glass rod to dissolve. Transfer the solution to a reagent bottle and bring the volume to 1.5 L. Cap the bottle and mix thoroughly by inverting the container. Mark the reagent name and preparation time with a marker.

[0043] 1 μM α-Lindenic acid treatment solution: Weigh 1 mg of α-Lindenic acid and dissolve it completely in 359.16 μL of DMSO to obtain a 10 mM stock solution; take 10 μL of the stock solution and add 9.99 mL of 5-methoxy-2-methylindole solution to obtain a final concentration of 10 μM dilution 1; take 10 mL of dilution 1 and add 90 mL of 5-methoxy-2-methylindole solution to obtain a 1 μM α-Lindenic acid treatment solution, and aliquot it into new 50 mL centrifuge tubes for later use.

[0044] Observe and record every 24 hours from the time different treatment solutions are added. Observe the time of appearance of transverse split bodies (early and late stages) and discoid bodies, the number of transverse split bodies, and the number of discoid bodies released per day. Throughout the experiment, pay close attention to the condition of the experimental animals, and the experiment ends when the last discoid body dies.

[0045] 3. Results Analysis

[0046] α-Lindenic acid inhibited the polyp metamorphosis of *A. coerulea* and reduced the number of discoid bodies released; the higher the concentration of α-Lindenic acid, the more significant the inhibitory effect. Experiments showed that, compared to the control group, 23.3% of individuals remained in the polyp stage four days after intervention with 1 μM α-Lindenic acid; and by days 7-8, most early transverse splits had not yet progressed to the next stage, and no discoid bodies were released. Figure 1 From the perspective of discoid body release, the number of discoid bodies released in the intervention group per day was significantly less than that in the control group. Furthermore, the intervention group released the most discoid bodies on days 9-10, while the control group showed the same effect on days 8-9, indicating a significantly delayed peak in discoid body release. Figure 2 In summary, α-Lindenic acid can inhibit the metamorphosis of A. coerulea hydroids and reduce the number of discoid bodies released; the higher the concentration of the inhibitor, the more significant the inhibitory effect.

[0047] Unless otherwise specified, the experimental methods used in the embodiments of this invention are conventional methods; unless otherwise specified, the materials and reagents used are commercially available.

Claims

1. Application of α-Lindenicacid in inhibiting the metamorphosis process of moon jellyfish.

2. The application according to claim 1, characterized in that, α-Lindenicacid has the following effects: (1) Delay the process of hydromorphic development; (2) Reduce the number of disc-shaped objects released; (3) Delay the time for discoid body release to reach its peak; (4) Delay the time when early transverse fractures begin to appear.

3. The application according to claim 2, characterized in that, The α-Lindenicacid was incubated together with the polyps of the moon jellyfish, and the concentration of α-Lindenicacid was 1 μM.

4. Application of α-Lindenicacid in the preparation of formulations that inhibit the metamorphosis of moon jellyfish.

5. The application according to claim 4, characterized in that, α-Lindenicacid has the following effects: (1) Delay the process of hydromorphic development; (2) Reduce the number of disc-shaped objects released; (3) Delay the time for discoid body release to reach its peak; (4) Delay the time when early transverse fractures begin to appear.

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