Method for evaluating oxidation level of coral lipid

Abstract

The invention relates to the technical field of biological health condition evaluation, in particular to a method for evaluating the oxidation level of coral lipid, which comprises the following steps: A) selecting a standard reference point: selecting hermatypic coral samples in an ideal growth state and under different stress conditions as reference samples, and detecting the malondialdehyde concentration in different reference samples; b) establishing a coral lipid oxidation level according to malondialdehyde concentration data of different reference samples and a coral growth state; and C) selecting coral to be evaluated as an experimental sample, detecting the concentration of malondialdehyde, and comparing the detection result of the experimental sample with the evaluation standard in the step B) to obtain the lipid oxidation level of the experimental sample. According to the method for evaluating the lipid oxidation level of the coral reef provided by the invention, the key technical problem of how to quickly evaluate the lipid oxidation level of hermatypic coral is solved, so that the coral reef is better protected and managed.

Description

Technical Field

[0001] The invention relates to the technical field of biological health status assessment, and in particular to a method for assessing coral lipid oxidation levels. Background Art

[0002] Coral reefs live in nutrient-poor waters and are known as the "tropical rainforests in the ocean". They have attracted much attention for their remarkable biodiversity and important economic value. Coral reefs have extraordinary primary productivity and have always been regarded as an indispensable key area in the ecosystem. Coral reefs not only have the ecological function of maintaining marine biodiversity, but also have ecological functions such as wave protection and bank protection and environmental regulation. At the same time, they contain rich resources and have important economic value. Its complex and diverse structure provides a unique habitat for various marine organisms, causing the entire ecosystem to show strong productivity. The main feature of the coral reef ecosystem is that its complex and varied reef structure provides a unique marine habitat for a wide variety of marine organisms, making the coral reef ecosystem highly productive.

[0003] Despite their many important values, coral reefs remain one of the most fragile ecosystems on Earth. In recent years, overfishing, land development and seawater pollution have been recognized as the main factors causing widespread changes in coral reef ecosystems. Large-scale coral bleaching and death not only cause devastating blows to the coral ecosystem itself, but also have a profound impact on the surrounding marine ecosystem and the marine economic development of human society. Therefore, effective protection and restoration measures must be taken as soon as possible to prevent irreversible damage to the ecosystem.

[0004] The marine environment is extremely complex and changeable. Slight fluctuations in factors such as temperature, salinity, and light may have an impact on corals. In the seabed environment, all types of corals will inevitably have to deal with environmental disturbances. The traditional method of assessing the health of corals is mainly to observe with the naked eye whether there are symptoms such as bleaching and tissue shedding on the surface of the corals to judge the health of the corals. However, once these phenomena occur, it often means that the corals are already in an extremely dangerous state and have missed the best time to save them. Therefore, there is an urgent need for a fast and easy-to-operate method to assess the level of coral lipid oxidation, so as to timely evaluate the recovery effect and stability of the coral reef ecosystem. Then, more scientific protection, restoration and management plans can be formulated to achieve sustainable use of coral reef resources. Summary of the invention

[0005] In view of this, the present invention provides a method for evaluating the lipid oxidation level of corals, which solves the key technical problem of how to quickly evaluate the lipid oxidation level of reef-building corals, so as to better protect and manage coral reefs.

[0006] The present invention provides a method for evaluating the lipid oxidation level of corals, comprising the following steps:

[0007] A) Select reef-building coral samples under ideal growth conditions and different stress conditions as reference samples, and detect the concentration of malondialdehyde in different reference samples;

[0008] B) Establish an evaluation standard for coral lipid oxidation level based on the malondialdehyde concentration data of different reference samples and the coral growth status;

[0009] C) Selecting the coral to be evaluated as the experimental sample, performing malondialdehyde concentration detection, comparing the detection result of the experimental sample with the evaluation standard in step B), and obtaining the evaluation result of the lipid oxidation level of the experimental sample.

[0010] Preferably, in step A), the reef-building coral sample is a reef-building coral, which is Acropora hyacinthoides or Acropora fasciata.

[0011] Preferably, in step A), the reef-building coral samples under different stress conditions are reef-building coral samples in a high temperature stress environment; the high temperature is 30-34°C.

[0012] Preferably, in step A), a malondialdehyde kit is used to detect the malondialdehyde concentrations in different reference samples.

[0013] Preferably, in step A), the method for detecting the concentration of malondialdehyde in different reference samples comprises the following steps:

[0014] Different samples were mixed with the extract respectively, homogenized in an ice bath, and centrifuged at 4°C. The obtained supernatant was used to measure the malondialdehyde concentration using a malondialdehyde kit.

[0015] Preferably, the extract is a phosphate buffered saline solution with a concentration of 0.08 to 0.12 mol / L.

[0016] Preferably, the dosage ratio of the sample to the extract is 0.08-0.12 g: 0.8-1.2 mL.

[0017] Preferably, step B) specifically comprises:

[0018] B1) Dividing the malondialdehyde concentration of each reference sample by the average malondialdehyde concentration of the reference samples to obtain a ratio of the reference sample value to the reference average value, as shown in formula (1):

[0019]

[0020] In formula (1), Y is the ratio of sample value to mean value;

[0021] X – MDA concentration of coral samples;

[0022] ——The average value of MDA concentration in each group of corals;

[0023] B2) Classifying the ratios in step B1) according to the growth conditions of the corals, thereby establishing a standard for evaluating the lipid oxidation level of the corals.

[0024] Preferably, step C) specifically comprises:

[0025] C1) Select the corals to be evaluated as experimental samples and test the concentration of malondialdehyde;

[0026] C2) according to formula (1), the malondialdehyde concentration of each experimental sample is divided by the average malondialdehyde concentration of the experimental samples to obtain the ratio of the experimental sample value to the experimental average value;

[0027] C3) comparing the ratio with the evaluation standard to obtain an evaluation result of the lipid oxidation level of the experimental sample.

[0028] Preferably, in step C1), the coral to be evaluated is a four-season clustered helmet coral, or the coral to be evaluated includes a natural Acropora hyacinthus and an artificial Acropora hyacinthus.

[0029] The present invention provides a method for evaluating the lipid oxidation level of corals, comprising the following steps: A) selecting a standard reference point: selecting reef-building coral samples under ideal growth conditions and under different stress conditions as reference samples, and detecting the malondialdehyde concentrations in different reference samples; B) establishing an evaluation standard for the lipid oxidation level of corals according to the malondialdehyde concentration data of different reference samples and the growth state of corals; C) selecting the corals to be evaluated as experimental samples, performing malondialdehyde concentration detection, and comparing the test results of the experimental samples with the evaluation standards in step B) to obtain the evaluation results of the lipid oxidation level of the experimental samples. The method for evaluating the lipid oxidation level of coral reefs provided by the present invention solves the key technical problem of how to quickly evaluate the health status of reef-building corals, thereby better protecting and managing coral reefs. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] Figure 1 A 3D view of a healthy cluster of helmet corals;

[0031] Figure 2 A 3D view of a sub-healthy cluster of helmet corals;

[0032] Figure 3 A 3D view of a bleached cluster of helmet corals;

[0033] Figure 4A 3D view of a healthy Acropora hyacinthus coral;

[0034] Figure 5 A three-dimensional view of a sub-healthy Acropora hyacinthus coral;

[0035] Figure 6 A three-dimensional view of a bleached Acropora hyacinthus coral;

[0036] Figure 7 A 3D view of a cluster of helmet corals in spring;

[0037] Figure 8 A three-dimensional view of a cluster of helmet corals in summer;

[0038] Fig. 9 A 3D view of a cluster of helmet corals in autumn;

[0039] Fig.10 A 3D view of a winter cluster of helmet corals. DETAILED DESCRIPTION

[0040] The technical solution of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0041] In the ocean, once environmental factors such as temperature, salinity, light and sediment change, corals will respond to relevant stresses and produce reactive oxygen species (ROS). The effects of ROS mainly include cell membrane lipid peroxidation, loss of organelle function, reduced metabolic efficiency, reduced carbon fixation and reduced photosynthesis, etc. Excessive ROS can cause oxidative damage and bleaching of coral tissues. Coral bleaching refers to the whitening of corals due to the loss of coral algae. Among them, malondialdehyde is one of the final products of lipid peroxidation, so malondialdehyde concentration is usually used as a marker of lipid peroxidation. Therefore, lipid peroxidation is believed to play a role in coral bleaching. Changes at the cellular and biochemical levels are detectable responses to environmental disturbances and can be observed before signs of physical stress such as bleaching are obvious. Therefore, malondialdehyde concentration is used as an indicator to judge whether corals are healthy or not.

[0042] The present invention provides a method for evaluating the lipid oxidation level of corals, comprising the following steps:

[0043] A) Select standard reference points: Select reef-building coral samples under ideal growth conditions and different stress conditions as reference samples, and detect the concentration of malondialdehyde in different reference samples;

[0044] B) Establish an evaluation standard for coral lipid oxidation level based on the malondialdehyde concentration data of different reference samples and the coral growth status;

[0045] C) Selecting the coral to be evaluated as the experimental sample, performing malondialdehyde concentration detection, comparing the detection result of the experimental sample with the evaluation standard in step B), and obtaining the evaluation result of the lipid oxidation level of the experimental sample.

[0046] Regarding step A):

[0047] Select standard reference points: Select reef-building coral samples under ideal growth conditions and under different stress conditions as reference samples, and detect the MDA concentration in different reference samples.

[0048] Malondialdehyde is produced when the lipids in the membranes of tissues or organs undergo oxidation reactions due to the aging of organisms or damage under adverse conditions. Its content is closely related to the aging of organisms and damage under adverse conditions. Therefore, the concentration of malondialdehyde is used as a biological indicator to judge the level of lipid oxidation in corals.

[0049] The reef-building coral samples under the ideal growth state refer to reef-building corals in the water temperature between 23℃ and 29℃, salinity between 32‰ and 35‰, and pH value between 8.1 and 8.5. Reef-building corals under such environmental conditions are closer to their ideal growth state, providing an important reference for studying their growth characteristics and ecological functions.

[0050] The reef-building coral sample is selected from a reef-building coral, which is Acropora hyacinthoides or Acropora fasciata. The number of coral samples can be 10.

[0051] The reef-building coral samples under different stress conditions can be reef-building coral samples in a high temperature stress environment. The high temperature can be 30-34°C, such as 32°C.

[0052] In certain embodiments of the present application, the malondialdehyde concentration in different reference samples is detected using a malondialdehyde (MDA) kit (TBA method).

[0053] Taking one of the samples as an example, the specific detection method includes the following steps:

[0054] The sample and the extract were mixed, homogenized in an ice bath, and centrifuged at 4°C. The supernatant was used to measure the malondialdehyde concentration using a malondialdehyde (MDA) kit (TBA method).

[0055] The extract is a phosphate buffered saline solution (PBS) with a concentration of 0.08-0.12 mol / L, such as 0.1 mol / L, obtained by mixing and diluting a 1 mol / L dipotassium hydrogen phosphate solution and a 1 mol / L potassium dihydrogen phosphate solution.

[0056] The dosage ratio of the sample to the extract is 0.08-0.12 g:0.8-1.2 mL, for example 0.1 g:1 mL.

[0057] The extracting solution used in the present invention can optimize the extraction process of malondialdehyde by maintaining a stable pH value, reducing the degradation of malondialdehyde, improving its solubility and reducing interference, thereby ensuring the reliability and repeatability of the experimental results.

[0058] The centrifugal speed is 12500-13000 rpm, such as 12000 rpm; the time is 8-12 min, such as 10 min.

[0059] Regarding step B):

[0060] Establishment of lipid oxidation level assessment criteria: Establishment of coral lipid oxidation level assessment criteria based on the malondialdehyde concentration data of different reference samples and the coral growth status.

[0061] Specifically, they include:

[0062] B1) Dividing the malondialdehyde concentration of each reference sample by the average malondialdehyde concentration of the reference samples to obtain a ratio of the reference sample value to the reference average value, as shown in formula (1):

[0063]

[0064] In formula (1), Y is the ratio of sample value to mean value;

[0065] X – MDA concentration of coral samples;

[0066] ——The average value of MDA concentration in each group of corals;

[0067] B2) Classifying the ratios in step B1) according to the growth conditions of the corals, thereby establishing a standard for evaluating the lipid oxidation level of the corals.

[0068] For example: the ratio corresponding to healthy corals is the first category, the ratio corresponding to sub-healthy corals is the second category, and the ratio corresponding to bleached corals is the third category.

[0069] Regarding step C):

[0070] C) Selecting the coral to be evaluated as the experimental sample, performing malondialdehyde concentration detection, comparing the detection result of the experimental sample with the evaluation standard in step B), and obtaining the evaluation result of the lipid oxidation level of the experimental sample.

[0071] Specifically, they include:

[0072] C1) Select the corals to be evaluated as experimental samples and test the concentration of malondialdehyde;

[0073] C2) according to formula (1), the malondialdehyde concentration of each experimental sample is divided by the average malondialdehyde concentration of the experimental samples to obtain the ratio of the experimental sample value to the experimental average value;

[0074] C3) comparing the ratio with the evaluation standard to obtain an evaluation result of the lipid oxidation level of the experimental sample.

[0075] Regarding step C1):

[0076] In certain embodiments of the present invention, the coral to be evaluated is a four-season clustered helmet coral, or the coral to be evaluated includes a natural Acropora hyacinthus and an artificial Acropora hyacinthus.

[0077] In certain embodiments of the present application, the malondialdehyde concentration in the experimental sample is detected using a malondialdehyde (MDA) kit (TBA method) in the same manner as the reference sample described above.

[0078] Taking one of the samples as an example, the specific detection method includes the following steps:

[0079] The sample and the extract were mixed, homogenized in an ice bath, and centrifuged at 4°C. The supernatant was used to measure the malondialdehyde concentration using a malondialdehyde (MDA) kit (TBA method).

[0080] In some embodiments, before mixing the sample with the extract, the process further includes: washing the sample with seawater to remove microorganisms, algae, sediments and other organic matter attached to the surface of the coral.

[0081] The extract is a phosphate buffered saline solution (PBS) with a concentration of 0.08-0.12 mol / L, such as 0.1 mol / L, obtained by mixing and diluting a 1 mol / L dipotassium hydrogen phosphate solution and a 1 mol / L potassium dihydrogen phosphate solution.

[0082] The dosage ratio of the sample to the extract is 0.08-0.12 g:0.8-1.2 mL, for example 0.1 g:1 mL.

[0083] The centrifugal speed is 12500-13000 rpm, such as 12000 rpm; the time is 8-12 min, such as 10 min.

[0084] Regarding step C3):

[0085] After comparing the ratio with the evaluation standard to obtain the evaluation result of the lipid oxidation level of the experimental sample, the growth condition of the coral surface is observed through a stereo microscope to judge the health status of the coral, thereby verifying the reliability of the evaluation standard.

[0086] Technical advantages and beneficial effects:

[0087] 1) Fast speed, simple experimental operation, can test about 100 samples in about 50 minutes;

[0088] 2) Malondialdehyde (MDA) assay kit (TBA method) The kit is easily available and affordable;

[0089] 3) Malondialdehyde (MDA) assay kit (TBA method) The kit has good stability and is valid for 12 months at 2-8°C;

[0090] 4) The color of the reagent is stable after reaction, and the absorbance value remains unchanged within two hours of color development;

[0091] 5) Low degree of external influence and high repeatability;

[0092] 6) It can more quickly assess the health of corals at a physiological level before the corals show visible bleaching, tissue shedding, etc., so as to carry out protection and restoration of coral reefs in a timely manner.

[0093] The present invention has no particular limitation on the sources of the raw materials used above, and they can be generally commercially available.

[0094] In order to further illustrate the present invention, a method for evaluating the lipid oxidation level of coral provided by the present invention is described in detail below in conjunction with examples, but it should not be construed as limiting the scope of protection of the present invention.

[0095] Figure 1 A 3D view of a healthy cluster of helmet corals. Figure 2 A 3D view of a sub-healthy cluster of helmet corals. Figure 3 A 3D view of a bleached cluster of helmet corals.

[0096] Figure 4 3D view of a healthy Acropora hyacinthus coral. Figure 5 A 3D view of a sub-healthy Acropora hyacinthus. Figure 6 A 3D view of a bleached Acropora hyacinthus coral.

[0097] Example 1

[0098] Taking the health status assessment of Acropora hyacinthus and Galaxea fascicularis as examples, the assessment criteria for coral lipid oxidation levels were established:

[0099] 1) Select 10 samples of Acropora hyacinthus and Acropora fasciata in ideal growth conditions, and 10 samples of Acropora hyacinthus and Acropora fasciata in 32°C high temperature stress environment, for a total of 40 samples. Detect the concentration of malondialdehyde in different samples, specifically, including:

[0100] Take 0.1g tissue sample from each sample, add 1mL 0.1mol / L phosphate buffered saline (PBS) extract (the extract is obtained by mixing and diluting 1mol / L dipotassium hydrogen phosphate solution and 1mol / L potassium dihydrogen phosphate solution), homogenize in an ice bath environment, centrifuge at 12000rpm for 10min at 4℃, and use the malondialdehyde (MDA) kit (TBA method) produced by Grease Biotechnology to measure the malondialdehyde concentration in the supernatant. The concentration unit is expressed in μmol / gprot. As shown in Table 1.

[0101] Table 1 Malondialdehyde concentrations of Acropora hyacinthoides and Galaxea fasciata in different health conditions

[0102]

[0103] 2) Establish evaluation criteria for coral lipid oxidation levels based on the malondialdehyde concentration data of different reference samples and the health status of corals:

[0104] 2-1) Group different types of corals, with each type of coral as one group;

[0105] 2-2) Divide each sample data of each group by the average value of the group to obtain the ratio of the reference sample value to the reference average value, as shown in formula (1):

[0106]

[0107] In formula (1), Y is the ratio of sample value to mean value;

[0108] X——malondialdehyde content of coral samples;

[0109] ——Average value of MDA content in corals of each group.

[0110] 2-3) According to the growth status of the coral, the ratios in step 2-2) are classified to establish the evaluation criteria for the lipid oxidation level of the coral:

[0111] The ratios of corals with low lipid oxidation levels were classified as the first category, the ratios of corals with intermediate lipid oxidation levels were classified as the second category, and the ratios of corals with high lipid oxidation levels were classified as the third category, as shown in Table 2.

[0112] Table 2 Health assessment criteria for Acropora hyacinthoides and Galaxea fasciata based on MDA concentration

[0113]

[0114] From Table 1 to 2, Figures 1 to 6 It can be seen that different stress environments have a significant impact on the health status and bleaching degree of different types of corals, and further lead to differences in their malondialdehyde concentrations. Therefore, the above data were used to establish an evaluation system for the lipid oxidation level of Acropora hyacinthoides and Galaxea fasciata based on malondialdehyde concentration.

[0115] Example 2

[0116] Galaxea fascicularis in four seasons were selected as experimental samples:

[0117] 1) Collected clumps of helmet corals from the wild, divided into 4 groups according to different seasons, 5 samples in each group, a total of 20 samples. After the coral samples were rinsed with seawater, 0.1g of tissue samples were cut, and 1mL of 0.1mol / L phosphate buffered saline (PBS) extract (the extract was obtained by mixing and diluting 1mol / L dipotassium hydrogen phosphate solution and 1mol / L potassium dihydrogen phosphate solution) was added. After homogenization in an ice bath environment, it was placed at 4°C and centrifuged at 12000rpm for 10min. The supernatant was used to measure the malondialdehyde concentration using the malondialdehyde (MDA) kit (TBA method) produced by Grace Biotechnology. The concentration unit is expressed in μmol / g prot. The test results are shown in Table 3.

[0118] 2) Calculate the ratio:

[0119] 2-1) Group different types of corals, with each type of coral as one group;

[0120] 2-2) According to formula (1), each experimental sample data of each group is divided by the experimental average value of the group to obtain the ratio of the experimental sample value to the experimental average value.

[0121] The ratio was compared with the evaluation standard of Example 1 to obtain the evaluation results of the lipid oxidation level of the experimental samples, as shown in Table 3.

[0122] Table 3 Evaluation results of lipid oxidation levels of experimental samples

[0123]

[0124] At the same time, the growth of coral surface was observed through stereo microscope to judge the lipid oxidation level of coral. The results are as follows: Figure 7 to Figure 10 shown. Figure 7 A 3D view of a cluster of helmet corals in spring. Figure 8 A 3D view of a cluster of helmet corals in summer. Fig. 9 A 3D view of a cluster of helmet corals in autumn. Fig.10 A 3D view of a winter cluster of helmet corals.

[0125] The experimental results show that the lipid oxidation levels presented by the stereomicroscope detection results are consistent with Table 3, thus verifying the reliability of the evaluation criteria in Table 2.

[0126] Example 3

[0127] The Acropora hyacinthoides in natural reef areas and Acropora hyacinthoides in artificial reef areas were selected as experimental samples:

[0128] 1) Acropora hyacinthoides samples were collected from natural reefs and artificial reefs, and divided into two groups according to environmental conditions. Each group contained 5 biological replicates, and a total of 10 samples were collected. After the coral samples were rinsed with seawater, 0.1g of tissue samples were cut and 1mL of 0.1mol / L phosphate buffered saline (PBS) extract (the extract was obtained by mixing and diluting 1mol / L dipotassium hydrogen phosphate solution and 1mol / L potassium dihydrogen phosphate solution) was added. After homogenization in an ice bath environment, it was placed at 4°C and centrifuged at 12000rpm for 10min. The supernatant was used to determine the malondialdehyde concentration using the malondialdehyde (MDA) kit (TBA method) produced by Grace Biotechnology. The concentration unit is expressed in μmol / g prot. As shown in Table 4.

[0129] 2) Calculate the ratio:

[0130] 2-1) Group different types of corals, with each type of coral as one group;

[0131] 2-2) According to formula (1), each experimental sample data of each group is divided by the experimental average value of the group to obtain the ratio of the experimental sample value to the experimental average value.

[0132] The ratio was compared with the evaluation standard of Example 1 to obtain the evaluation results of the lipid oxidation level of the experimental samples, as shown in Table 4.

[0133] Table 4 Evaluation results of lipid oxidation levels of experimental samples

[0134] area Natural reef area Artificial reef area MDA concentration 23.18±4.91 23.92±4.58 ratio 0.98 1.02 Lipid oxidation level Low high

[0135] At the same time, the growth conditions of the coral surface were observed through a stereo microscope to judge the health of the corals. The results showed that the health level presented by the stereo microscope test results was consistent with Table 4, thus verifying the reliability of the evaluation criteria in Table 2.

[0136] From Table 3, Figures 7-10 It can be seen that there are significant differences in the concentration of MDA in the clump of helmet corals in different seasons, especially in summer, which is significantly higher than the other three groups, slightly lower in autumn, and no significant difference between spring and winter. The results in Table 4 show that there is no significant difference in the concentration of MDA in the healthy state of Acropora hyacinthoides in different environments, with high consistency. Therefore, MDA concentration can be used as a new method to assess the health status of corals.

[0137] The above results show that the coral lipid oxidation level assessment method using MDA as an evaluation indicator is highly scientific and reliable. This method can monitor the health of corals in a timely manner, has important guiding significance for the protection and restoration of coral reefs, and helps promote the long-term development of my country's coral reef ecosystem protection work.

[0138] The description of the above embodiments is only used to help understand the method of the present invention and its core idea. Various modifications to these embodiments will be apparent to those skilled in the art, and the general principles defined herein can be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but will conform to the widest range consistent with the principles and novel features disclosed herein.

Claims

1. A method for assessing the level of lipid oxidation in corals, comprising the following steps: A) Select reef-building coral samples under ideal growth conditions and different stress conditions as reference samples, and detect the concentration of malondialdehyde in different reference samples; B) Establish an evaluation standard for coral lipid oxidation level based on the malondialdehyde concentration data of different reference samples and the coral growth status; C) Selecting the coral to be evaluated as the experimental sample, performing malondialdehyde concentration detection, comparing the detection result of the experimental sample with the evaluation standard in step B), and obtaining the evaluation result of the lipid oxidation level of the experimental sample.

2. The method for evaluating lipid oxidation level according to claim 1, characterized in that: In step A), the reef-building coral sample is selected from a reef-building coral, which is Acropora hyacinthoides or Acropora fasciata.

3. The method for evaluating lipid oxidation level according to claim 1, characterized in that: In step A), the reef-building coral samples under different stress conditions are reef-building coral samples in a high temperature stress environment; the high temperature is 30-34°C.

4. The method for evaluating lipid oxidation level according to claim 1, characterized in that: In step A), the malondialdehyde concentration in different reference samples is detected using a malondialdehyde kit.

5. The method for evaluating lipid oxidation level according to claim 1, characterized in that: In step A), the method for detecting the concentration of malondialdehyde in different reference samples comprises the following steps: Different samples were mixed with the extract respectively, homogenized in an ice bath, and centrifuged at 4°C. The obtained supernatant was used to measure the malondialdehyde concentration using a malondialdehyde kit.

6. The method for evaluating lipid oxidation level according to claim 5, characterized in that: The extract is a phosphate buffered saline solution with a concentration of 0.08-0.12 mol / L.

7. The method for evaluating lipid oxidation level according to claim 5, characterized in that: The usage ratio of the sample to the extract is 0.08-0.12 g: 0.8-1.2 mL.

8. The method for evaluating lipid oxidation level according to claim 1, characterized in that: Step B) specifically includes: B1) Dividing the malondialdehyde concentration of each reference sample by the average malondialdehyde concentration of the reference samples to obtain a ratio of the reference sample value to the reference average value, as shown in formula (1): In formula (1), Y is the ratio of sample value to mean value; X – MDA concentration of coral samples; ——The average value of MDA concentration in each group of corals; B2) Classifying the ratios in step B1) according to the growth conditions of the corals, thereby establishing a standard for evaluating the lipid oxidation level of the corals.

9. The method for evaluating lipid oxidation level according to claim 1, characterized in that: Step C) specifically comprises: C1) Select the corals to be evaluated as experimental samples and test the concentration of malondialdehyde; C2) according to formula (1), the malondialdehyde concentration of each experimental sample is divided by the average malondialdehyde concentration of the experimental samples to obtain the ratio of the experimental sample value to the experimental average value; C3) comparing the ratio with the evaluation standard to obtain an evaluation result of the lipid oxidation level of the experimental sample.

10. The method for evaluating lipid oxidation level according to claim 9, characterized in that: In step C1), the coral to be evaluated is a four-season clustered helmet coral, or the coral to be evaluated includes a natural Acropora hyacinthus and an artificial Acropora hyacinthus.

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