Microelement supplement as well as preparation method, application and use method thereof

Abstract

The invention provides a trace element supplement as well as a preparation method, application and a use method thereof, and particularly relates to the technical field of coral culture. The microelement supplement consists of a cation microelement supplement and an anion microelement supplement, the cation trace element supplement is prepared from SrCl2, Fe-EDTA (Ethylene Diamine Tetraacetic Acid), Mn-EDTA, Zn-EDTA, Ni-EDTA, BaCl2, CuCl2, CoCl2, LiCl and distilled water; the anion trace element supplement is prepared from NaBr, H3BO3, Na2MoO4, KI, KF, Na2CrO4 and distilled water. According to the microelement supplement, the microelement pedigree in the natural growth environment of the coral is reduced through the collaborative design of an anion-cation system. The supplement mainly supplements key elements which are easy to lack in an artificial breeding system, so that effective coverage of microelement requirements of coral in the whole growth cycle is realized.

Description

Technical Field

[0001] The present invention relates to the technical field of coral cultivation, and particularly to a trace element supplement, a preparation method thereof, an application, and a usage method. Background Art

[0002] As an important part of the marine ecosystem, corals not only provide diverse habitats for marine organisms but also greatly enrich the biodiversity of the marine ecosystem. The growth of corals depends on the mutualistic symbiotic relationship with symbiotic algae - zooxanthellae. The algae provide essential organic matter for corals through photosynthesis, while corals provide a habitat and carbon dioxide for the algae. The color manifestation of corals mainly stems from the pigment differences within the symbiotic algae. Different species of zooxanthellae affect the color display effect of corals due to their unique pigment components. In addition, the fluorescent proteins and pigment proteins synthesized by corals themselves also participate in their color presentation, and the expression levels of these proteins are often closely related to the environmental conditions and environmental pressures during the growth process of corals.

[0003] With the increasing prominence of the ornamental value of corals and the continuous improvement of cultivation techniques, coral cultivation, as a technology for artificially breeding and cultivating corals in a controlled environment, has seen a significant increase in its proportion in the commercial aquarium market. Under the ideal environmental conditions of natural waters, corals usually exhibit good growth states and rich and plump colors. However, it is often difficult to fully replicate the complexity of natural ecosystems in artificial cultivation environments, resulting in many restrictions on coral growth, among which the lack of trace elements in the water body is particularly prominent. Especially in closed aquariums or cultivation systems, the concentrations of minerals and trace elements often cannot fully meet the physiological needs of corals and their symbiotic algae.

[0004] Currently, the coral cultivation field in China faces a shortage of trace element supplement products. Existing products mainly consist of single-element supplements or solid powder formulations. Single-element supplementation easily leads to the problem of unbalanced trace element intake by corals during cultivation, while simply using multiple elements in combination may cause adverse reactions such as precipitation and complexation due to improper ratios. In addition, solid powder supplements generally have technical bottlenecks in poor solubility in actual cultivation environments, and these problems urgently need to be solved through product research and development and technological innovation.

[0005] In view of this, the present invention is specifically proposed. Summary of the Invention

[0006] One of the purposes of the present invention is to provide a trace element supplement, aiming to solve at least one of the above technical problems in the prior art.

[0007] Another purpose of the present invention is to provide a preparation method of the trace element supplement.

[0008] A third object of the present invention is to provide an application of a trace element supplement.

[0009] A fourth object of the present invention is to provide a method for using a trace element supplement.

[0010] In order to achieve the above objects of the present invention, the following technical solutions are specifically adopted:

[0011] The first aspect of the present invention provides a trace element supplement, which is composed of a cationic trace element supplement and an anionic trace element supplement.

[0012] The cationic trace element supplement includes SrCl 2 , Fe-EDTA, Mn-EDTA, Zn-EDTA, Ni-EDTA, BaCl 2 , CuCl 2 , CoCl 2 , LiCl and distilled water.

[0013] The anionic trace element supplement includes NaBr, H 3 BO 3 , Na 2 MoO 4 , KI, KF, Na 2 CrO 4 and distilled water.

[0014] Further, by weight, the cationic trace element supplement includes SrCl 2 50-160 parts, Fe-EDTA 3-18 parts, Mn-EDTA 1-3 parts, Zn-EDTA 0.3-1.5 parts, Ni-EDTA 0.01-0.08 parts, BaCl 2 0.3-1.5 parts, CuCl 2 0.1-0.8 parts, CoCl 2 0.001-0.02 parts, LiCl 0.5-2 parts and distilled water 1000 parts.

[0015] Further, by weight, the anionic trace element supplement includes NaBr 90-160 parts, H 3 BO 3 15-35 parts, Na 2 MoO 4 0.2-1.2 parts, KI 0.8-1.8 parts, KF 5-25 parts, Na 2 CrO 4 0.001-0.007 parts and distilled water 1000 parts.

[0016] Further, by weight parts, the cationic trace element supplement includes SrCl 2 70 - 150 parts, Fe-EDTA 5 - 15 parts, Mn-EDTA 1 - 2 parts, Zn-EDTA 0.5 - 1 part, Ni-EDTA 0.01 - 0.05 part, BaCl 2 0.5 - 1 part, CuCl 2 0.1 - 0.5 part, CoCl 2 0.001 - 0.01 part and LiCl 1 - 1.5 parts.

[0017] Further, by weight parts, the anionic trace element supplement includes NaBr 100 - 150 parts, H 3 BO 3 20 - 30 parts, Na 2 MoO 4 0.4 - 1 part, KI 1 - 1.5 parts, KF 6 - 20 parts and Na 2 CrO 4 0.001 - 0.005 part.

[0018] The second aspect of the present invention provides a preparation method of the trace element supplement, including the following steps:

[0019] A. Add SrCl 2 , Fe-EDTA, Mn-EDTA, Zn-EDTA, Ni-EDTA, BaCl 2 , CuCl 2 , CoCl 2 and LiCl into distilled water and mix evenly until a clear solution is obtained to get the cationic trace element supplement;

[0020] B. Add NaBr, H 3 BO 3 , Na 2 MoO 4 , KI, KF and Na 2 CrO 4 into distilled water and mix evenly until a clear solution is obtained to get the anionic trace element supplement.

[0021] The third aspect of the present invention provides an application of the trace element supplement in coral cultivation.

[0022] Further, the coral includes Acropora.

[0023] The fourth aspect of the present invention provides a usage method of the trace element supplement. During the coral cultivation process, the addition amount of the trace element supplement is 0.1 - 0.2 g / kg of water.

[0024] Furthermore, the time interval between the addition of the cationic trace element supplement and the anionic trace element supplement is greater than 1 hour.

[0025] Compared with the prior art, the present invention has at least the following beneficial effects:

[0026] The trace element supplement provided by the present invention, through the collaborative design of the cation-anion system, successfully restores the trace element spectrum in the natural growth environment of corals. This supplement specifically focuses on supplementing the key elements that are easily lacking in artificial aquaculture systems, thereby effectively covering the trace element requirements of corals throughout the growth cycle. All trace elements exist in a safe form, without introducing harmful impurity ions, and are stable in the product, with high adaptability to different water qualities and strong biological availability. This trace element supplement can significantly increase the growth rates of the wet weight, volume, and density of corals, while promoting the expression level of common fluorescent protein genes in corals and enhancing the pigment synthesis ability of corals. In addition, this supplement can also effectively promote the color development of corals themselves and their symbiotic zooxanthellae, further enhancing the expression level of coral fluorescent protein genes and accelerating pigment synthesis, so that the artificially cultured corals are more brightly colored and the ornamental value is significantly improved.

[0027] The preparation method of the trace element supplement provided by the present invention has a simple process and can be prepared without special equipment, making it suitable for large-scale industrial production.

[0028] Regarding the application of the trace element supplement provided by the present invention, due to the advantages possessed by the above trace element supplement, it provides an efficient, safe, and environmentally friendly solution for the coral aquaculture industry. At the same time, this supplement has broad application prospects in the fields of coral reef restoration, marine ecological restoration, and high-end ornamental coral aquaculture.

[0029] The usage method of the trace element supplement provided by the present invention can accurately meet the trace element requirements of corals at different growth stages by controlling the addition amount of the trace element supplement within the range of 0.1 - 0.2 g / kg of water, avoiding growth restrictions caused by insufficient addition or element concentration fluctuations caused by excessive addition, thereby ensuring the healthy growth of corals. This usage method is simple and easy to implement, without the need for complex equipment or frequent water quality monitoring, reducing the technical threshold of coral aquaculture and being suitable for large-scale promotion and application. BRIEF DESCRIPTION OF THE DRAWINGS

[0030] In order to more clearly illustrate the specific embodiments of the present invention or the technical solutions in the prior art, the following will briefly introduce the drawings required for the description of the specific embodiments or the prior art. Obviously, the drawings in the following description are some embodiments of the present invention. For those of ordinary skill in the art, without creative efforts, other drawings can also be obtained based on these drawings.

[0031] Figure 1 It is a graph of the detection result of coral growth state;

[0032] Figure 2 It is a graph of the detection result of coral pigment content;

[0033] Figure 3 It is a graph of the qPCR detection result of coral. Specific implementation manners

[0034] To make the objectives, technical solutions and advantages of the present invention clearer, the technical solutions in the embodiments 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 a part rather than all of the embodiments of the present invention.

[0035] In the following text, the terms "including", "having" and their cognates that can be used in various embodiments of the present invention are only intended to represent specific features, numbers, steps, operations, elements, components or combinations of the foregoing items, and should not be construed as first excluding the existence of one or more other features, numbers, steps, operations, elements, components or combinations of the foregoing items or increasing the possibility of one or more features, numbers, steps, operations, elements, components or combinations of the foregoing items.

[0036] The first aspect of the present invention provides a trace element supplement, which is composed of a cationic trace element supplement and an anionic trace element supplement.

[0037] The cationic trace element supplement includes SrCl 2 , Fe-EDTA, Mn-EDTA, Zn-EDTA, Ni-EDTA, BaCl 2 , CuCl 2 , CoCl 2 , LiCl and distilled water.

[0038] The anionic trace element supplement includes NaBr, H 3 BO 3 , Na 2 MoO 4 , KI, KF, Na 2 CrO 4 and distilled water.

[0039] The trace element supplement provided by the present invention successfully restores the trace element spectrum in the natural growth environment of corals through the collaborative design of the cation-anion system. This supplement focuses on replenishing the key elements that are easily lacking in artificial breeding systems, thereby effectively covering the trace element requirements of corals throughout their growth cycle. All trace elements exist in a safe form, without introducing harmful impurity ions, and are stable in the product, with high adaptability to different water qualities and strong biological availability. This trace element supplement can significantly increase the growth rates of the wet weight, volume, and density of corals, while promoting the expression levels of common fluorescent protein genes in corals and enhancing the pigment synthesis ability of corals. In addition, this supplement can effectively promote the color development of corals themselves and their symbiotic zooxanthellae, further enhancing the expression levels of coral fluorescent protein genes and accelerating pigment synthesis, thereby making the artificially cultured corals more colorful and significantly improving their ornamental value.

[0040] Further, by weight parts, the cationic trace element supplement includes SrCl 2 50 - 160 parts, Fe-EDTA 3 - 18 parts, Mn-EDTA 1 - 3 parts, Zn-EDTA 0.3 - 1.5 parts, Ni-EDTA 0.01 - 0.08 parts, BaCl 2 0.3 - 1.5 parts, CuCl 2 0.1 - 0.8 parts, CoCl 2 0.001 - 0.02 parts, LiCl 0.5 - 2 parts and 1000 parts of distilled water.

[0041] Typical but non-limiting, in the cationic trace element supplement, the weight parts of SrCl 2 can be 50 parts, 60 parts, 70 parts, 80 parts, 90 parts, 100 parts, 110 parts, 120 parts, 130 parts, 140 parts, 150 parts or 160 parts, or any value within the range of 50 - 160 parts; the weight parts of Fe-EDTA can be 3 parts, 6 parts, 9 parts, 12 parts, 15 parts or 18 parts, or any value within the range of 3 - 18 parts; the weight parts of Mn-EDTA can be 1 part, 1.5 parts or 3 parts, or any value within the range of 1 - 3 parts; the weight parts of Zn-EDTA can be 0.3 parts, 0.6 parts, 0.9 parts, 1.2 parts or 1.5 parts, or any value within the range of 0.3 - 1.5 parts; the weight parts of Ni-EDTA can be 0.01 parts, 0.02 parts, 0.04 parts, 0.06 parts or 0.08 parts, or any value within the range of 0.01 - 0.08 parts; the weight parts of BaCl 2 can be 0.3 parts, 0.6 parts, 0.9 parts, 1.2 parts or 1.5 parts, or any value within the range of 0.3 - 1.5 parts; CuCl2 The weight parts thereof can be 0.1 part, 0.3 part, 0.5 part, 0.6 part, 0.7 part or 0.8 part, or can also be any value within the range of 0.1 to 0.8 parts; CoCl 2 The weight parts thereof can be 0.001 part, 0.005 part, 0.01 part, 0.015 part or 0.02 part, or can also be any value within the range of 0.001 to 0.02 parts; the weight parts of LiCl can be 0.5 part, 1 part, 1.5 parts or 2 parts, or can also be any value within the range of 0.5 to 2 parts.

[0042] Further, by weight parts, the anionic trace element supplement includes 90 to 160 parts of NaBr, H 3 BO 3 15 to 35 parts, Na 2 MoO 4 0.2 to 1.2 parts, KI 0.8 to 1.8 parts, KF 5 to 25 parts, Na 2 CrO 4 0.001 to 0.007 parts and 1000 parts of distilled water.

[0043] Typical but non-limiting, in the anionic trace element supplement, the weight parts of NaBr can be 90 parts, 100 parts, 110 parts, 120 parts, 130 parts, 140 parts, 150 parts or 160 parts, or can also be any value within the range of 90 to 160 parts; H 3 BO 3 The weight parts thereof can be 15 parts, 20 parts, 25 parts, 30 parts or 35 parts, or can also be any value within the range of 15 to 35 parts; Na 2 MoO 4 The weight parts thereof can be 0.2 part, 0.4 part, 0.6 part, 0.8 part, 1.0 part, 1.2 parts, or can also be any value within the range of 0.2 to 1.2 parts; the weight parts of KI can be 0.8 part, 1.0 part, 1.2 parts, 1.4 parts, 1.6 parts or 1.8 parts, or can also be any value within the range of 0.8 to 1.8 parts; the weight parts of KF can be 5 parts, 10 parts, 15 parts, 20 parts, 25 parts, or can also be any value within the range of 5 to 25 parts; Na 2 CrO 4 The weight parts thereof can be 0.001 part, 0.002 part, 0.003 part, 0.004 part, 0.005 part, 0.006 part or 0.007 part, or can also be any value within the range of 0.001 to 0.007 parts.

[0044] Further, by weight parts, the cationic trace element supplement includes SrCl 270 to 150 parts, Fe-EDTA 5 to 15 parts, Mn-EDTA 1 to 2 parts, Zn-EDTA 0.5 to 1 part, Ni-EDTA 0.01 to 0.05 parts, BaCl 2 0.5 to 1 part, CuCl 2 0.1 to 0.5 part, CoCl 2 0.001 to 0.01 part and LiCl 1 to 1.5 parts.

[0045] Further, by weight parts, the anion trace element supplement includes NaBr 100 to 150 parts, H 3 BO 3 20 to 30 parts, Na 2 MoO 4 0.4 to 1 part, KI 1 to 1.5 parts, KF 6 to 20 parts and Na 2 CrO 4 0.001 to 0.005 part.

[0046] The second aspect of the present invention provides a preparation method of the trace element supplement described above, including the following steps:

[0047] A. Add SrCl 2 , Fe-EDTA, Mn-EDTA, Zn-EDTA, Ni-EDTA, BaCl 2 , CuCl 2 , CoCl 2 and LiCl into distilled water and mix evenly until a clear solution is obtained to get the cation trace element supplement;

[0048] B. Add NaBr, H 3 BO 3 , Na 2 MoO 4 , KI, KF and Na 2 CrO 4 into distilled water and mix evenly until a clear solution is obtained to get the anion trace element supplement.

[0049] The preparation method of the trace element supplement provided by the present invention has a simple process and can be prepared without special equipment, which is suitable for large-scale industrial production.

[0050] The third aspect of the present invention provides an application of the trace element supplement described above in coral cultivation.

[0051] The application of the trace element supplement provided by the present invention, in view of the advantages of the above-mentioned trace element supplement, provides an efficient, safe and environmentally friendly solution for the coral aquaculture industry. At the same time, the supplement has broad application prospects in the fields of coral reef restoration, marine ecological restoration and high-end ornamental coral aquaculture.

[0052] Further, the coral includes Acropora.

[0053] The fourth aspect of the present invention provides a method for using the trace element supplement. During the coral aquaculture process, the addition amount of the trace element supplement is 0.1-0.2 g / kg of water.

[0054] The method for using the trace element supplement provided by the present invention can accurately meet the trace element requirements of corals at different growth stages by controlling the addition amount of the trace element supplement within the range of 0.1-0.2 g / kg of water, avoiding growth limitation caused by insufficient addition amount or element concentration fluctuations caused by excessive addition, thereby ensuring the healthy growth of corals. This method is simple and easy to implement, without the need for complex equipment or frequent water quality monitoring, reducing the technical threshold of coral aquaculture and being suitable for large-scale promotion and application.

[0055] In the specific use process, the addition amount of the trace element supplement per kilogram of water can be, for example, 0.1 g, 0.12 g, 0.14 g, 0.16 g, 0.18 g or 0.2 g, or any value within the range of 0.1-0.2 g.

[0056] Further, the time interval between the addition of the cationic trace element supplement and the anionic trace element supplement is greater than 1 hour.

[0057] The present invention requires a strategy of adding the cationic and anionic trace element supplements at an interval of more than 1 hour, which significantly improves the efficiency of coral aquaculture through multi-dimensional synergistic effects. This operation first avoids the precipitation reaction caused by the direct contact of cations (such as Sr 2+ , Ba 2+ ) and anions (such as MoO 4 2- , CrO 4 2- ), and optimizes the bioavailability of elements by improving the stability of EDTA chelates; secondly, it matches the absorption timing of the coral calcification layer and symbiotic algae to improve the ion transmembrane transport efficiency; at the same time, it maintains the stability of the water quality, reduces the pH fluctuation range, and avoids I - oxidation loss.

[0058] The present invention will be further described below through specific examples and comparative examples. However, it should be understood that these examples are only for more detailed illustration and should not be construed as limiting the present invention in any way. For the raw materials used in the examples and comparative examples of the present invention, unless otherwise specified, they are carried out under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used without indicating the manufacturer are all conventional products that can be obtained through commercial purchase.

[0059] Example 1

[0060] This example provides a trace element supplement, and its raw materials include the following components in parts by weight: Cationic trace element supplement: SrCl 2 ·6H 2 O 120 parts, Fe-EDTA (chemical formula: C 10 H 12 FeN 2 NaO 8 ) 10 parts, Mn-EDTA (chemical formula: C 10 H 12 MnN 2 NaO 8 ) 1.5 parts, Zn-EDTA (chemical formula: C 10 H 12 ZnN 2 NaO 8 ) 0.7 part, BaCl 2 ·2H 2 O 1 part, CuCl 2 ·2H 2 O 0.1 part, Ni-EDTA (chemical formula: C 10 H 12 NiN 2 NaO 8 ·xH 2 O) 0.03 part, CoCl 2 ·6H 2 O 0.007 part, LiCl 1.2 parts and 1000 parts of distilled water.

[0061] Anionic trace element supplement: NaBr 130 parts, H 3 BO 3 29 parts, Na 2 MoO 4 ·2H 2 O 0.5 part, KI 1.2 parts, KF·2H 2 O 10 parts, Na 2 CrO 4 ·4H 2 O 0.004 part and 1000 parts of distilled water.

[0062] The preparation method of the trace element supplement in this embodiment includes the following steps:

[0063] Weigh each component of the cationic trace element supplement according to the formula, dissolve it in distilled water, and stir well to form a clear solution without precipitates, thus obtaining the cationic trace element supplement. Weigh each component of the anionic trace element supplement according to the formula, dissolve it in distilled water, and stir well to form a clear solution without precipitates, thus obtaining the anionic trace element supplement.

[0064] Example 2

[0065] This example provides a trace element supplement, and its raw materials include the following components in parts by weight:

[0066] Cationic trace element supplement: SrCl 2 ·6H 2 O 110 parts, Fe-EDTA (chemical formula: C 10 H 12 FeN 2 NaO 8 ) 5 parts, Mn-EDTA (chemical formula: C 10 H 12 MnN 2 NaO 8 ) 1 part, Zn-EDTA (chemical formula: C 10 H 12 ZnN 2 NaO 8 ) 0.5 part, BaCl 2 ·2H 2 O 0.5 part, CuCl 2 ·2H 2 O 0.1 part, Ni-EDTA (chemical formula: C 10 H 12 NiN 2 NaO 8 ·xH 2 O) 0.01 part, CoCl 2 ·6H 2 O 0.001 part, LiCl 1 part and 1000 parts of distilled water.

[0067] Anionic trace element supplement: NaBr 100 parts, H 3 BO 3 20 parts, Na 2 MoO 4 ·2H 2 O 0.5 part, KI 1 part, KF·2H 2 O 10 parts, Na 2 CrO4 ·4H 2 0.001 parts and 1000 parts of distilled water.

[0068] The preparation method is the same as that of Example 1 and will not be repeated here.

[0069] Example 3

[0070] This example provides a trace element supplement, and its raw materials include the following components in parts by weight:

[0071] Cationic trace element supplement: SrCl 2 ·6H 2 O 150 parts, Fe-EDTA (chemical formula: C 10 H 12 FeN 2 NaO 8 ) 15 parts, Mn-EDTA (chemical formula: C 10 H 12 MnN 2 NaO 8 ) 2 parts, Zn-EDTA (chemical formula: C 10 H 12 ZnN 2 NaO 8 ) 1 part, BaCl 2 ·2H 2 O 1 part, CuCl 2 ·2H 2 O 0.5 part, Ni-EDTA (chemical formula: C 10 H 12 NiN 2 NaO 8 ·xH 2 O) 0.05 part, CoCl 2 ·6H 2 O 0.01 part, LiCl 1.5 parts and 1000 parts of distilled water.

[0072] Anionic trace element supplement: NaBr 150 parts, H 3 BO 3 30 parts, Na 2 MoO 4 ·2H 2 O 1 part, KI 1.5 parts, KF·2H 2 O 20 parts, Na 2 CrO 4 ·4H 2 O 0.005 part and 1000 parts of distilled water.

[0073] The preparation method is the same as that of Example 1 and will not be repeated here.

[0074] Comparative Example 1

[0075] This comparative example provides a trace element supplement, which is different from Example 1 in that it only contains cationic trace element supplements and does not include anionic trace element supplements. The remaining raw materials and preparation methods are the same as those in Example 1 and will not be elaborated here.

[0076] Comparative Example 2

[0077] This comparative example provides a trace element supplement, which is different from Example 1 in that it only contains anionic trace element supplements and does not include cationic trace element supplements. The remaining raw materials and preparation methods are the same as those in Example 1 and will not be elaborated here.

[0078] Comparative Example 3

[0079] In this comparative example, coral cultivation is carried out without adding trace element supplements.

[0080] Experimental Example

[0081] The trace element supplements provided in the examples and comparative examples are used for coral cultivation.

[0082] I. Experimental Process

[0083] 1. Experimental object: The experimental coral is Acropora (Acropora, Acroporidae, Scleractinia, Hexacorallia, Cnidaria).

[0084] 2. Experimental environment: The experimental corals are placed in an aquarium for cultivation. The specifications of the aquarium are 120 cm × 60 cm × 40 cm. Six aquariums are selected, and 3 corals are placed in each aquarium. Before the formal experiment, in order to enable the corals to adapt to the cultivation environment, the corals are first domestically raised for 14 days, and then healthy and similar corals are selected as experimental strains for grouping experiments according to the experimental requirements.

[0085] 3. Experimental grouping: The trace element supplements described in the present invention are used for artificial cultivation of the corals in the examples (EG) and comparative examples (CG) for 49 days.

[0086] 4. Experimental period: The experimental period is 49 days. During this process, according to a mass ratio of 1:10,000, 0.2 g of trace element supplements (the mass ratio of anionic trace element supplements to cationic trace element supplements is 1:1) are added to the aquarium every 7 days, and the time interval between adding cationic trace element supplements and anionic trace element supplements is greater than 1 h. Moreover, 20% of the water in the aquarium is changed every 7 days.

[0087] 5. Growth state detection: On the 0th day and the 49th day of the experiment, the total wet weight of corals in each group was weighed using an analytical balance, the volume of corals in each group was measured by the drainage method, and the average density of corals in each group was calculated. The results are shown in Table 1 and Figure 1 as follows.

[0088] 6. Pigment content detection: The contents of chlorophyll and carotenoids in the corals of the experimental examples and comparative examples were detected by colorimetry. On a clean workbench, an appropriate amount of coral tissue samples were taken and placed in a mortar, and a small amount of quartz sand, calcium carbonate powder and 95% ethanol were added and ground into a homogenate. After standing for 5 min, the grinding liquid was filtered, and all the carotenoids on the filter paper were washed into ethanol and made up to a constant volume. The extract was poured into a 1 cm optical path cuvette, and 95% ethanol was used as the blank, and the absorbance was measured at wavelengths of 665 nm, 649 nm and 470 nm. The detection results are shown in Table 2 and Figure 2 as follows.

[0089] 7. Detection of the expression level of fluorescent protein gene: The expression level of fluorescent protein gene in the corals of the experimental examples and comparative examples was detected by real-time fluorescence quantitative PCR (qPCR). On a clean workbench, an appropriate amount of coral tissue samples were taken, RNA was extracted using Trizol reagent, and the concentration and purity of RNA were measured using NanoDrop 2000 for RNA quality detection. The qualified and quantified total RNA was reverse transcribed into cDNA using a reverse transcription kit (PrimeScriptTM 1st stand cDNA Synthesis Kit). The PCR reaction system was configured and the reaction was carried out on a Realtime PCR instrument. According to the obtained fluorescence signal curve, parameters such as Ct value were analyzed to detect the expression level of fluorescent protein gene in the corals. The results are shown in Table 5 and Figure 3 as follows.

[0090] II. Analysis of detection results

[0091] 1. Data analysis of growth state detection

[0092] Table 1 Detection results of coral growth state

[0093]

[0094] As shown in Table 1, after 49 days of cultivation with the trace element supplement, the three growth state detection indexes of average wet weight, average volume and average density of the corals in the examples were significantly improved. Among them, the average wet weight of the corals in Example 1 increased by 302%, the average volume increased by 25%, and the average density increased by 221.6%. Figure 1It shows that for the corals cultured with trace element supplements for 49 days, the average wet weight, average volume and average density growth rates of the corals in the examples are significantly higher than those in the comparative examples (p≤0.05). Among them, the effect of Example 1 is the best. In summary, the trace element supplement can promote the growth of corals and improve the growth state of corals.

[0095] 2. Data analysis of pigment content detection

[0096] Table 2 Detection results of coral pigment content

[0097]

[0098] As shown in Table 2, after the corals in the examples were cultured with trace element supplements for 49 days, the average contents of chlorophyll a, chlorophyll b and carotenoids increased significantly compared with those in the comparative examples. Among them, the average content of chlorophyll a in the corals of Example 1 was 0.029 mg / g, the average content of chlorophyll b was 0.022 mg / g, and the average content of carotenoids was 0.006 mg / g. Figure 2 It shows that for the corals cultured with trace element supplements for 49 days, the chlorophyll a content and carotenoid content of the corals in the examples are significantly higher than those in the comparative examples (p≤0.05), and the content of chlorophyll b is generally higher than that of the corals in the comparative examples but not significantly (p>0.05). In summary, the trace element supplement can improve the pigment content synthesized by corals.

[0099] 3. Data analysis of qPCR detection

[0100] Table 3 Selected coral fluorescent protein genes and internal reference genes

[0101]

[0102]

[0103] Table 4 Primer information for qPCR detection

[0104]

[0105] Table 5 qPCR reaction system

[0106] Reagent System 2×SYBR real-time PCR premixture 10μl PCR specific primer F at 10uM 0.4μl PCR specific primer R at 10uM 0.4μl cDNA 1μl <![CDATA[RNase free dH 2 O]]> Up to 20μl

[0107] To quantitatively explore the effect of trace element supplements on the color development of corals, the expression levels of 7 common fluorescent protein genes in corals were detected, and the internal reference gene actin was selected for normalization. The gene sequences of the fluorescent protein genes and the internal reference gene can be found on the website of the National Center for Biotechnology Information (NCBI), as shown in Table 3. Specific primers were designed (as shown in Table 4), and a qPCR reaction system was constructed (as shown in Table 5) for real-time fluorescence quantitative PCR detection. The 2 -ΔΔCT -method was used to calculate the gene expression levels, and the detection results are shown in Table 6 and Figure 3 .

[0108] Table 6 Coral qPCR Detection Results

[0109]

[0110] As shown in Table 6, after 49 days of cultivation with trace element supplements, the expression levels of the 7 common fluorescent protein genes in the corals of the examples all increased significantly. Figure 3 This indicates that the trace element supplements have a significant promoting effect on the color development of corals. Among them, the expression levels of 6 fluorescent protein genes, namely Adi-Fluorescent protein-1, amilCFP, amilGFP, Adi-Fluorescent protein-10, amilRFP, and Adi-Fluorescent protein-7, in the corals of the examples were significantly higher than those of the comparative examples (p≤0.05). Only the detection result of the Adi-Fluorescent protein-2 gene was not significant (p>0.05), but the expression level of this gene was still generally higher than that of the comparative examples in the examples. In summary, trace element supplements have obvious benefits for the color development of corals.

[0111] Finally, it should be noted that the above-described embodiments are only specific embodiments of the present invention, used to illustrate the technical solutions of the present invention, rather than limiting them. The protection scope of the present invention is not limited thereto. Although the present invention has been described in detail with reference to the foregoing embodiments, those of ordinary skill in the art should understand that: any person skilled in the art within the technical scope disclosed by the present invention can still modify the technical solutions recorded in the foregoing embodiments or can easily think of changes, or perform equivalent replacements on some of the technical features; and these modifications, changes or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present invention, and should all be covered by the protection scope of the present invention. Therefore, the protection scope of the present invention should be subject to the protection scope of the claims.

Claims

1. A trace element supplement, characterized in that: It is composed of cationic trace element supplement and anionic trace element supplement; The cationic trace element supplement includes SrCl2, Fe-EDTA, Mn-EDTA, Zn-EDTA, Ni-EDTA, BaCl2, CuCl2, CoCl2, LiCl and distilled water; The anion trace element supplement includes NaBr, H3BO3, Na2MoO4, KI, KF, Na2CrO4 and distilled water.

2. The trace element supplement according to claim 1, characterized in that: Calculated by weight, the cationic trace element supplement includes 50-160 parts of SrCl2, 3-18 parts of Fe-EDTAA, 1-3 parts of Mn-EDTAA, 0.3-1.5 parts of Zn-EDTAA, 0.01-0.08 parts of Ni-EDTAA, 0.3-1.5 parts of BaCl2, 0.1-0.8 parts of CuCl2, 0.001-0.02 parts of CoCl2, 0.5-2 parts of LiCl and 1000 parts of distilled water.

3. The trace element supplement according to claim 1, characterized in that: Calculated by weight, the anionic trace element supplement includes 90-160 parts of NaBr, 15-35 parts of H3BO3, 0.2-1.2 parts of Na2MoO4, 0.8-1.8 parts of KI, 5-25 parts of KF, 0.001-0.007 parts of Na2CrO4 and 1000 parts of distilled water.

4. The trace element supplement according to claim 1, characterized in that: Calculated by weight, the cationic trace element supplement includes 70-150 parts of SrCl2, 5-15 parts of Fe-EDTA, 1-2 parts of Mn-EDTA, 0.5-1 part of Zn-EDTA, 0.01-0.05 part of Ni-EDTAA, 0.5-1 part of BaCl2, 0.1-0.5 part of CuCl2, 0.001-0.01 part of CoCl2 and 1-1.5 parts of LiCl.

5. The trace element supplement according to claim 1, characterized in that: Calculated by weight, the anionic trace element supplement includes 100-150 parts of NaBr, 20-30 parts of H3BO3, 0.4-1 part of Na2MoO4, 1-1.5 parts of KI, 6-20 parts of KF and 0.001-0.005 parts of Na2CrO4.

6. A method for preparing the trace element supplement according to any one of claims 1 to 5, characterized in that: The following steps are involved: A. Add SrCl2, Fe-EDTA, Mn-EDTA, Zn-EDTA, Ni-EDTA, BaCl2, CuCl2, CoCl2 and LiCl into distilled water and mix well until a clear solution is obtained to obtain a cationic trace element supplement; B. Add NaBr, H3BO3, Na2MoO4, KI, KF and Na2CrO4 into distilled water and mix well until a clear solution is obtained to obtain an anionic trace element supplement.

7. Use of the trace element supplement according to any one of claims 1 to 5 in coral cultivation.

8. The use according to claim 7, characterized in that: The corals include Acropora.

9. A method for using the trace element supplement according to any one of claims 1 to 5, characterized in that: During the coral cultivation process, the trace element supplement is added in an amount of 0.1 to 0.2 g / kg water.

10. The method of use according to claim 9, characterized in that: The time interval between the addition of cationic trace element supplement and anionic trace element supplement is greater than 1 hour.

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