Method for inducing ostrea rivularis triploid

Abstract

The invention discloses a method for inducing ostrea rivularis triploid, and relates to the technical field of shellfish genetic breeding. The specific preparation method comprises the following steps: firstly, obtaining egg liquid and seminal fluid from gonads of female and male ostrea rivularis individuals, cleaning and curing the egg liquid, and adding the seminal fluid into the egg liquid after cleaning to obtain a fertilized egg solution; when the proportion of the first polar bodies in the eggs of the fertilized egg solution is 30-50%, adding an inducer into the fertilized egg solution for induction and cleaning to obtain induced fertilized eggs; and finally, hatching and screening the induced fertilized eggs to obtain D-shaped larvae, and culturing to obtain the crassostrea rivularis triploid population. The method for inducing the crassostrea rivularis triploid is high in triploid induction rate, high in final hatching rate, capable of stably surviving and good in application prospect.

Description

Technical Field

[0001] The invention relates to the technical field of shellfish genetic breeding, and in particular to a method for inducing triploid Omi oysters. Background Art

[0002] Oysters are the most widely cultivated and productive marine shellfish in my country. The Omi oyster belongs to the phylum Mollusca, class Bivalvia, order Pectinatus, family Ostreidae. It is the most widely distributed oyster in my country. It is a euryhaline species with low-salinity tolerance and usually lives in estuaries with low salinity.

[0003] However, there are certain problems in the cultivation of Omi oysters, such as the poor taste and low meat yield of diploid Omi oysters, as well as the frequent large-scale death during the cultivation process, which causes economic losses. Therefore, a production method is urgently needed to solve this problem. Although the performance of Omi oyster triploids is poor, they can improve the meat yield of Omi oysters, have a high survival rate, and meet the needs of the market throughout the year, so they have broad market prospects. In order to obtain commercial Omi oyster triploids, it is necessary to first induce the production of Omi oyster triploids. Only by inducing the production of Omi oyster triploids can we induce the production of Omi oyster tetraploids through the induced triploids in the future and then cultivate commercial Omi oyster triploids. Summary of the invention

[0004] The purpose of the invention is to provide a method for inducing triploid Crassostrea gigas to solve the problems of low survival rate and poor taste of existing diploid Crassostrea gigas.

[0005] In order to solve the above technical problems, the present invention specifically provides the following technical solutions: The invention discloses a method for inducing triploidy of Crassostrea gigas, comprising: S1: The gonads of male and female individuals of Omi oysters are peeled and crushed in seawater to obtain egg liquid and semen, the egg liquid is washed and immersed in seawater to mature to obtain mature egg liquid, and the semen is washed to obtain pure semen; then, the pure semen is added to the mature egg liquid to obtain a fertilized egg solution; S2: When the proportion of the first polar body in the eggs in the fertilized egg solution is 30-50%, an induction agent is added to the fertilized egg solution for induction, and finally the induced fertilized eggs are obtained by washing; S3: The induced fertilized eggs are placed in seawater for hatching, and D-shaped larvae are screened after hatching; the D-shaped larvae are placed in seawater for cultivation, and finally transferred to an outdoor cement pool for growth.

[0006] Preferably, the temperature of the seawater in S1 is 20-30°C, and the salinity of the seawater is 15-20ppt.

[0007] Preferably, the ratio of the number of eggs in the matured egg fluid to the number of sperms in the pure semen in S1 is 1:10-15.

[0008] Preferably, the egg liquid in S1 is cleaned with a 200-550 mesh silk sieve; the semen is cleaned with a 250-350 mesh silk sieve.

[0009] Preferably, the inducer in S2 is at least one of cytochalasin B and a purine derivative, and the amount of the inducer in the fertilized egg solution is 0.4-8 mg / L; the purine derivative is prepared from 2-chloro-6-(4-morpholinyl)-9H-purine, ethyl 2-(bromomethyl)thiazole-5-carboxylate and 4-(ethylenesulfonyl)-aniline. The use of purine derivatives can more effectively inhibit microtubule growth and interfere with polar bodies, and the induction effect is stable, thereby improving the triploid induction rate of Omi oysters and obtaining a stable and high hatching rate.

[0010] Preferably, the cleaning reagent in S2 is a dimethyl sulfoxide solution, which is composed of dimethyl sulfoxide and water, and the volume ratio of dimethyl sulfoxide to water is 1:900-1100.

[0011] Preferably, the temperature of the seawater in S3 is 20-30°C, and the salinity of the seawater is 15-20ppt.

[0012] Preferably, the sieve used in S3 is a 250-350 mesh sieve.

[0013] Preferably, the immersion time in seawater in S1 is 25-40 min; and the induction time in S2 is 15-20 min.

[0014] Preferably, during the incubation process in S3, micro-aeration is continuously performed, stirring is performed every 1.5-2.5 hours, and the incubation time is 20-30 hours.

[0015] The invention discloses a method for inducing triploidy of Crassostrea gigas, comprising: S1: The gonads of male and female individuals of Omi oysters are peeled and crushed in seawater to obtain egg liquid and semen, the egg liquid is washed and immersed in seawater for 25-40 minutes to obtain mature egg liquid, and the semen is washed to obtain pure semen; then, the pure semen is added to the mature egg liquid to obtain a fertilized egg solution; S2: When the proportion of the first polar body in the eggs in the fertilized egg solution is 30-50%, an induction agent is added to the fertilized egg solution for induction for 15-20 minutes, and finally washed to obtain the induced fertilized eggs; S3: Place the induced fertilized eggs in seawater for incubation, continuously aerate during the incubation process, stir every 1.5-2.5 hours, and screen for D-shaped larvae after 20-30 hours of incubation. Place the D-shaped larvae in seawater, feed them every 3-5 hours, and change the seawater 1-3 times a day. When the proportion of eyed larvae in the D-shaped larvae reaches 25-35%, place the attachment substrate, and cultivate for another 18-23 days, and finally transfer them to an outdoor cement pool for growth.

[0016] Preferably, the temperature of the seawater in S1 is 20-30°C, and the salinity of the seawater is 15-20ppt.

[0017] Preferably, the ratio of the number of eggs in the matured egg fluid to the number of sperms in the pure semen in S1 is 1:10-15.

[0018] Preferably, the egg liquid in S1 is cleaned with a 200-550 mesh silk sieve; the semen is cleaned with a 250-350 mesh silk sieve.

[0019] Preferably, the inducer in S2 is at least one of cytochalasin B and a purine derivative, and the amount of the inducer in the fertilized egg solution is 0.4-8 mg / L.

[0020] Preferably, the cleaning reagent in S2 is a dimethyl sulfoxide solution, which is composed of dimethyl sulfoxide and water, and the volume ratio of dimethyl sulfoxide to water is 1:900-1100.

[0021] Preferably, the temperature of the seawater in S3 is 20-30°C, and the salinity of the seawater is 15-20ppt.

[0022] Preferably, the sieve used in S3 is a 250-350 mesh sieve.

[0023] Preferably, the density of D-larvae in S3 in seawater is 8-12 / ml.

[0024] Preferably, the bait used for feeding in S3 is Isochrysis galbana, and the daily feeding amount of the bait is 8000-30000 cells / mL.

[0025] Preferably, the amount of seawater replaced each time in S3 is 45-55% of the original seawater.

[0026] The present invention discloses a method for preparing a purine derivative, which specifically comprises: 2-Chloro-6-(4-morpholinyl)-9H-purine, ethyl 2-(bromomethyl)thiazole-5-carboxylate and potassium carbonate were added to N,N-dimethylformamide and reacted at 20-30°C for 4-7h. After the reaction, distillation and extraction were performed, and the organic phase was collected, and then washed and dried to obtain an intermediate. The intermediate and 4-(ethylenesulfonyl)-aniline were added to methanol and reacted at 65-80°C for 4-7h. After the reaction, distillation and extraction were performed, and the organic phase was collected, and then washed and dried to obtain a crude product. The crude product was purified by column chromatography to obtain a purine derivative.

[0027] Preferably, the mass ratio of 2-chloro-6-(4-morpholinyl)-9H-purine to ethyl 2-(bromomethyl)thiazole-5-carboxylate is 1:1.2-2.2.

[0028] Preferably, the mass ratio of 2-chloro-6-(4-morpholinyl)-9H-purine to potassium carbonate is 1:3-4.2.

[0029] Preferably, the usage ratio of 2-chloro-6-(4-morpholinyl)-9H-purine to N,N-dimethylformamide is 1 g:14-15 ml.

[0030] Preferably, the mass ratio of the intermediate to 4-(ethylenesulfonyl)-aniline is 1:0.6-1.2.

[0031] Preferably, the usage ratio of the intermediate and methanol is 1 g: 30-50 ml.

[0032] Preferably, the reagents used for extraction are water and ethyl acetate.

[0033] Preferably, the reagents used for washing are all saturated saline.

[0034] More preferably, in the method of inducing triploidy in Omi oysters, in addition to using purine derivatives, berberine hydrochloride hydrate can also be used. The synergistic use of berberine hydrochloride hydrate and purine derivatives can play a synergistic role, further promote the interference with polar bodies, and improve the induction stability, thereby further improving the triploid induction rate and hatching rate of Omi oysters.

[0035] Preferably, the amount of berberine hydrochloride hydrate in the fertilized egg solution is 0.5-1.2 mg / L.

[0036] Compared with the prior art, the present invention has the following beneficial effects: The present invention proposes a method for inducing triploid Omi oysters. First, egg liquid and semen are obtained from the gonads of male and female Omi oysters, the egg liquid is washed and matured, and the semen is washed and added to the egg liquid to obtain a fertilized egg solution; when the proportion of the first polar body in the eggs of the fertilized egg solution is 30-50%, an inducer is added to the fertilized egg solution to induce and wash to obtain induced fertilized eggs; finally, the induced fertilized eggs are hatched and screened to obtain D-shaped larvae, and a triploid population of Omi oysters is obtained through cultivation. The method for inducing triploid Omi oysters of the present invention has a high triploid induction rate and a high final hatching rate, and can survive stably, and has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0037] In order to more clearly illustrate the implementation methods of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for the implementation methods or the description of the prior art. Obviously, the drawings in the following description are only exemplary, and for ordinary technicians in this field, other implementation drawings can be derived from the provided drawings without creative work.

[0038] Figure 1 This is the result of the triploid induction rate determination; Figure 2 This is a graph showing the hatchability test results. DETAILED DESCRIPTION

[0039] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in 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.

[0040] The following first describes the concepts involved in the present application in conjunction with the accompanying drawings. It should be noted that the following description of each concept is only to make the content of the present application easier to understand, and does not limit the scope of protection of the present application; at the same time, the embodiments and features in the embodiments of the present application can be combined with each other in the absence of conflict. The present application will be described in detail with reference to the accompanying drawings and in conjunction with the embodiments.

[0041] Embodiment 1: Fertilization of Omi oysters: Select male and female individuals with full gonads from Omi oysters. Peel off the gonads of female individuals, crush them in seawater to obtain egg liquid, wash the egg liquid with 260 mesh silk sieve and 500 mesh silk sieve in turn, and then immerse it in seawater for 30 minutes to obtain mature egg liquid. Peel off the gonads of male individuals, crush them in seawater to obtain semen, and wash the semen with 300 mesh silk sieve to obtain pure semen. Add pure semen to the mature egg liquid to obtain fertilized egg solution. The temperature of seawater is 23℃, the salinity of seawater is 18ppt, and the ratio of the number of eggs in the mature egg liquid to the number of sperm in the pure semen is 1:12.

[0042] Induction of triploid in Omi oyster: When the proportion of the first polar body in the eggs in the fertilized egg solution is 40%, cytochalasin B is added to the fertilized egg solution for induction for 18 minutes. After the induction, the induced fertilized eggs are washed with dimethyl sulfoxide solution. The amount of cytochalasin B in the fertilized egg solution is 0.5 mg / L, and the dimethyl sulfoxide solution is composed of dimethyl sulfoxide and water, and the volume ratio of dimethyl sulfoxide to water is 1:1000.

[0043] Hatching and cultivation of Omi oysters: The induced fertilized eggs are placed in seawater for incubation. During the incubation process, they are continuously slightly aerated and stirred every 2 hours. After incubation for 24 hours. After the incubation, D-shaped larvae are screened with a 300-mesh silk sieve to obtain the D-shaped larvae. The D-shaped larvae are placed in seawater for cultivation. During the cultivation period, they are fed with Isochrysis globosum every 4 hours, and the seawater is changed twice a day. When the proportion of eye-spot larvae in the D-shaped larvae reaches 30%, the attachment substrate is placed, and the culture is continued for 20 days, and then transferred to an outdoor cement pool for cultivation. The seawater temperature is 23°C, the salinity of the seawater is 18ppt, the density of the D-shaped larvae in the seawater is 10 / ml, the daily feeding amount of Isochrysis globosum is 10,000 cells / mL, and the amount of seawater replaced each time is 50% of the original seawater volume.

[0044] Embodiment 2: The fertilization of Omi oysters was the same as in Example 1.

[0045] Preparation of purine derivatives: 2-chloro-6-(4-morpholinyl)-9H-purine, ethyl 2-(bromomethyl)thiazole-5-carboxylate and potassium carbonate were added to N,N-dimethylformamide and reacted at 25°C for 5 hours. After the reaction, distillation was performed, and water and ethyl acetate were added for extraction. The organic phase was collected, washed with saturated brine, and finally dried to obtain an intermediate. The intermediate and 4-(ethylenesulfonyl)-aniline were added to methanol, reacted at 70°C for 5 hours, distilled after the reaction, and then water and ethyl acetate were added for extraction. The organic phase was collected, washed with saturated brine, and finally dried to obtain a crude product. The crude product was purified by column chromatography to obtain a purine derivative. The mass ratio of 2-chloro-6-(4-morpholinyl)-9H-purine to ethyl 2-(bromomethyl)thiazole-5-carboxylate is 1:1.8, the mass ratio of 2-chloro-6-(4-morpholinyl)-9H-purine to potassium carbonate is 1:3.6, the mass ratio of 2-chloro-6-(4-morpholinyl)-9H-purine to N,N-dimethylformamide is 1g:14.67ml, the mass ratio of the intermediate to 4-(ethylenesulfonyl)-aniline is 1:0.98, and the mass ratio of the intermediate to methanol is 1g:40ml.

[0046] Induction of triploid in Omi oyster: When the proportion of the first polar body in the eggs in the fertilized egg solution is 40%, cytochalasin B and purine derivatives are added to the fertilized egg solution for induction for 18 minutes. After the induction, the induced fertilized eggs are washed with dimethyl sulfoxide solution. The amount of cytochalasin B in the fertilized egg solution is 0.5 mg / L, the amount of purine derivatives in the fertilized egg solution is 2 mg / L, the dimethyl sulfoxide solution is composed of dimethyl sulfoxide and water, and the volume ratio of dimethyl sulfoxide to water is 1:1000.

[0047] Hatching and cultivation of Omi oysters: The hatching and cultivation of Omi oysters in this embodiment is different from that in Example 1 in that the induced fertilized eggs are the induced fertilized eggs prepared in this embodiment, and the other conditions and parameters are the same as in Example 1.

[0048] Embodiment 3: The fertilization of Omi oysters was the same as in Example 1.

[0049] The preparation of purine derivatives is the same as in Example 2.

[0050] Induction of triploid Crassostrea gigas in this example: The induction of triploid Crassostrea gigas in this example is different from that in Example 2 in that the amount of purine derivatives in the fertilized egg solution is 5 mg / L, and other conditions and parameters are the same as in Example 2.

[0051] Hatching and cultivation of Omi oysters: The hatching and cultivation of Omi oysters in this embodiment is different from that in Example 2 in that the induced fertilized eggs are the induced fertilized eggs prepared in this embodiment, and the other conditions and parameters are the same as in Example 2.

[0052] Embodiment 4: The fertilization of Omi oysters was the same as in Example 1.

[0053] The preparation of purine derivatives is the same as in Example 2.

[0054] Induction of triploid Crassostrea gigas in this example: The induction of triploid Crassostrea gigas in this example is different from that in Example 2 in that the amount of purine derivatives in the fertilized egg solution is 1.2 mg / L, and other conditions and parameters are the same as in Example 2.

[0055] Hatching and cultivation of Omi oysters: The hatching and cultivation of Omi oysters in this embodiment is different from that in Example 2 in that the induced fertilized eggs are the induced fertilized eggs prepared in this embodiment, and the other conditions and parameters are the same as in Example 2.

[0056] Embodiment 5: The fertilization of Omi oysters was the same as in Example 1.

[0057] The preparation of purine derivatives is the same as in Example 2.

[0058] Induction of triploid in Omi oyster: When the proportion of the first polar body in the eggs of the fertilized egg solution is 40%, cytochalasin B, purine derivatives and berberine hydrochloride hydrate are added to the fertilized egg solution for induction for 18 minutes. After the induction, the induced fertilized eggs are washed with dimethyl sulfoxide solution to obtain the induced fertilized eggs. The amount of cytochalasin B in the fertilized egg solution is 0.5 mg / L, the amount of purine derivatives in the fertilized egg solution is 2 mg / L, the amount of berberine hydrochloride hydrate in the fertilized egg solution is 0.6 mg / L, and the dimethyl sulfoxide solution is composed of dimethyl sulfoxide and water, and the volume ratio of dimethyl sulfoxide to water is 1:1000.

[0059] Hatching and cultivation of Omi oysters: The hatching and cultivation of Omi oysters in this embodiment is different from that in Example 2 in that the induced fertilized eggs are the induced fertilized eggs prepared in this embodiment, and the other conditions and parameters are the same as in Example 2.

[0060] Embodiment 6: The fertilization of Omi oysters was the same as in Example 1.

[0061] The preparation of purine derivatives is the same as in Example 2.

[0062] Induction of triploid Crassostrea gigas in this example: The induction of triploid Crassostrea gigas in this example is compared with that in Example 5, except that the amount of berberine hydrochloride hydrate in the fertilized egg solution is 1 mg / L, and the other conditions and parameters are the same as in Example 5.

[0063] Hatching and cultivation of Omi oysters: The hatching and cultivation of Omi oysters in this embodiment is different from that in Example 5 in that the induced fertilized eggs are the induced fertilized eggs prepared in this embodiment, and the other conditions and parameters are the same as in Example 5.

[0064] Comparative Example 1: The fertilization of Omi oysters was the same as in Example 1.

[0065] The preparation of purine derivatives is the same as in Example 2.

[0066] Induction of triploid Crassostrea gigas in this comparative example was compared with that in Example 2, except that the amount of purine derivatives in the fertilized egg solution was 0.15 mg / L, and other conditions and parameters were the same as in Example 2.

[0067] Hatching and cultivation of Omi oysters: The hatching and cultivation of Omi oysters in this comparative example is compared with that in Example 2, except that the induced fertilized eggs are the induced fertilized eggs prepared in this example, and the other conditions and parameters are the same as in Example 2.

[0068] Experimental Example 1: Determination of the triploid induction rate: The D-shaped larvae obtained in Examples 1-6 and Comparative Example 1 were subjected to ploidy detection, and the triploid induction rate was calculated based on the ploidy. The triploid induction rate = triploid ratio / (triploid ratio + diploid ratio) × 100%.

[0069] The ploidy of the D-shaped larvae obtained in Examples 1-6 and Comparative Example 1 was detected, and the triploid induction rate was calculated as follows: Figure 1As shown. Compared with Example 2, Example 1 shows that the use of purine derivatives can improve the triploid induction rate of Omi oysters; compared with Example 3, Example 2 shows that the increase in the amount of purine derivatives used within a certain range can improve the triploid induction rate of Omi oysters; compared with Example 4, Example 2 shows that the reduction in the amount of purine derivatives used within a certain range will reduce the triploid induction rate of Omi oysters; compared with Example 5, Example 2 shows that on the basis of using purine derivatives, the use of berberine hydrochloride hydrate can further improve the triploid induction rate of Omi oysters; compared with Example 6, Example 5 shows that the increase in the amount of berberine hydrochloride hydrate used within a certain range can also improve the triploid induction rate of Omi oysters; compared with Comparative Example 1, Example 2 shows that the amount of purine derivatives used needs to be within an appropriate range. If the amount is too low, it will have no obvious effect on improving the triploid induction rate of Omi oysters.

[0070] Experimental Example 2: The hatching rate was determined by counting the number of fertilized eggs in the fertilized egg solutions obtained in Examples 1-6 and Comparative Example 1, and the number of D-shaped larvae finally hatched and cultured. The hatching rate = the number of D-shaped larvae / the number of fertilized eggs × 100%.

[0071] The hatching rate was calculated by counting the number of fertilized eggs and D-larvae obtained in Examples 1-6 and Comparative Example 1. Figure 2 As shown. Compared with Example 2, Example 1 shows that the use of purine derivatives can improve the hatching rate of D-shaped larvae; compared with Example 3, Example 2 shows that the increase in the amount of purine derivatives used within a certain range can improve the hatching rate of D-shaped larvae; compared with Example 4, Example 2 shows that the reduction in the amount of purine derivatives used within a certain range will reduce the hatching rate of D-shaped larvae; compared with Example 5, Example 2 shows that on the basis of using purine derivatives, the use of berberine hydrochloride hydrate can further improve the hatching rate of D-shaped larvae; compared with Example 6, Example 5 shows that the increase in the amount of berberine hydrochloride hydrate used within a certain range can also improve the hatching rate of D-shaped larvae; compared with Comparative Example 1, Example 2 shows that the amount of purine derivatives used needs to be within an appropriate range. If the amount is too low, it will have no obvious effect on improving the hatching rate of D-shaped larvae.

[0072] The embodiments and / or implementation methods described above are only used to illustrate the preferred embodiments and / or implementation methods for realizing the technology of the present invention, and are not intended to limit the implementation methods of the technology of the present invention in any form. Any technical personnel in this field may make slight changes or modifications to other equivalent embodiments without departing from the scope of the technical means disclosed in the content of the present invention, but they should still be regarded as technologies or embodiments that are essentially the same as the present invention.

[0073] This article uses specific examples to illustrate the principles and implementation methods of this application. The description of the above embodiments is only used to help understand the method and its core ideas of this application. The above is only the preferred implementation method of this application. It should be pointed out that due to the limitations of textual expression and the objective existence of infinite specific structures, ordinary technicians in this technical field can make several improvements, modifications or changes without departing from the principles of this application, and can also combine the above technical features in an appropriate manner; these improvements, modifications, changes or combinations, or the direct application of the concept and technical solution of the invention to other occasions without improvement, should be regarded as the scope of protection of this application.

Claims

1. A method for inducing triploidy in Crassostrea gigas, comprising: S1: The gonads of male and female individuals of Omi oysters are peeled and crushed in seawater to obtain egg liquid and semen, the egg liquid is washed and immersed in seawater to mature to obtain mature egg liquid, and the semen is washed to obtain pure semen; then, the pure semen is added to the mature egg liquid to obtain a fertilized egg solution; S2: When the proportion of the first polar body in the eggs in the fertilized egg solution is 30-50%, an induction agent is added to the fertilized egg solution for induction, and finally the induced fertilized eggs are obtained by washing; S3: The induced fertilized eggs are placed in seawater for hatching, and D-shaped larvae are screened after hatching; the D-shaped larvae are placed in seawater for cultivation, and finally transferred to an outdoor cement pool for growth.

2. The method for inducing triploidy of Crassostrea gigas according to claim 1, characterized in that: The temperature of the seawater in S1 is 20-30°C, and the salinity of the seawater is 15-20ppt.

3. The method for inducing triploidy of Crassostrea gigas according to claim 1, characterized in that: The ratio of the number of eggs in the matured egg fluid to the number of sperms in the pure semen in the S1 is 1:10-15.

4. The method for inducing triploidy of Crassostrea gigas according to claim 1, characterized in that: In the S1, the egg liquid is cleaned with a 200-550 mesh silk sieve; the semen is cleaned with a 250-350 mesh silk sieve.

5. The method for inducing triploidy of Crassostrea gigas according to claim 1, characterized in that: The inducer in S2 is at least one of cytochalasin B and a purine derivative, and the amount of the inducer in the fertilized egg solution is 0.4-8 mg / L; the purine derivative is prepared from 2-chloro-6-(4-morpholinyl)-9H-purine, 2-(bromomethyl)thiazole-5-carboxylic acid ethyl ester and 4-(ethylenesulfonyl)-aniline.

6. The method for inducing triploidy of Crassostrea gigas according to claim 1, characterized in that: The cleaning reagent in S2 is a dimethyl sulfoxide solution, which is composed of dimethyl sulfoxide and water. The volume ratio of dimethyl sulfoxide to water is 1:900-1100.

7. The method for inducing triploidy of Crassostrea gigas according to claim 1, characterized in that: The temperature of the seawater in S3 is 20-30°C, and the salinity of the seawater is 15-20ppt.

8. The method for inducing triploidy of Crassostrea gigas according to claim 1, characterized in that: The sieve used in S3 is a 250-350 mesh sieve.

9. The method for inducing triploidy of Crassostrea gigas according to claim 1, characterized in that: The immersion time of S1 in seawater is 25-40 minutes; the induction time of S2 is 15-20 minutes.

10. The method for inducing triploidy of Crassostrea gigas according to claim 1, characterized in that: During the incubation process in S3, micro-aeration is continuously performed, stirring is performed every 1.5-2.5 hours, and the incubation time is 20-30 hours.

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