A method of delaying spawning of reef fish gametes
By cleaning, air-drying, and low-temperature dark preservation of reef gametophytes, combined with soaking treatment under specific conditions, the problem of delayed gametophyte release in reef cultivation was solved, achieving effective control of gametophyte viability and quantity, thus meeting production needs.
Patent Information
- Application Number
- CN202411744263.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- Filing Date
- 2024-11-30
- Publication Date
- 2026-01-06
- Estimated Expiration
- 2044-11-30
AI Technical Summary
During reef membrane cultivation, existing technologies cannot effectively delay the release of gametophytes, leading to production stagnation or hindering collection, and no relevant research has been reported in the current field.
The gamete release was delayed by cleaning and air-drying the collected robust, mature, and undischarged reef gametes at room temperature, storing them in sealed bags at 4°C in the dark, and then immersing them in disinfected seawater under specific temperature, humidity, and light intensity conditions.
This method preserves gametophyte viability and delays gamete emission, meeting production needs, shortening the summering time of sporophytes, and reducing labor intensity and costs.
Abstract
Description
Technical Field
[0001] This invention belongs to the field of reproductive regulation technology in the cultivation of large-scale economic seaweed, specifically relating to a method for delaying the release of gametes from reef gametophytes. Background Technology
[0002] Monostroma nitidum is a large, economically important green algae with large, nutrient-rich gametophytes, possessing high economic value for both food and medicinal purposes. When the gametophyte matures, it forms gametocysts. After the gametes are released, they attach to the substrate, germinate, and grow into small sporophytes. Once mature, the sporophytes release spores, which then germinate and grow into gametophytes.
[0003] Reef gametophytes are the seed algae used for reef seedling cultivation. In the production practice of reef cultivation, the timing, speed and quantity of gamete release are key factors affecting production. Gamete release is affected by ecological factors such as water temperature, salinity and light intensity. Under suitable conditions, gamete release is concentrated and large in quantity; conversely, release is dispersed, small in quantity or even non-existent.
[0004] In the practice of reef membrane seedling cultivation, due to unfavorable weather or human factors, although the reef membrane gametophytes are mature, the conditions for gamete collection are not met, and gamete collection needs to be delayed. At this time, the mature gametophytes need to be treated to prevent the gametophytes from releasing gametes too early, otherwise the release of gametes will lead to production stagnation. After the unfavorable factors are removed, the gametes of the reef membrane need to be collected normally. Whether the treated mature gametophytes can release gametes normally is related to whether production can proceed smoothly.
[0005] In addition, in production, reef gametes are usually collected, attached to a substrate, and cultured into sporophytes, which then undergo summer cultivation. Delaying the release of reef gametes, thereby postponing their collection, can effectively shorten the summer cultivation time of reef sporophytes, reduce the labor intensity of summer management, and lower the cost of cultivation.
[0006] Currently, no research has been reported on the delayed reef gametophyte emission of gametes. Summary of the Invention
[0007] In view of this, the main objective of the present invention is to provide a method for delaying the release of gametes from reef gametophytes.
[0008] To achieve the objective, the technical solution of this invention is implemented as follows:
[0009] This invention provides a method for delaying reef membrane gamete discharge, the method comprising:
[0010] Step (1): At room temperature, place the collected robust, mature, and unreleased reef gametophytes on a clean workbench for cleaning and air drying;
[0011] Step (2): The algae from step (1) are placed in a sealed bag and stored at 4°C in the dark. Then, they are soaked in disinfected seawater at 90 to 900 times the mass of the algae and allowed to stand under the conditions of temperature, humidity and light intensity before the gametes are released.
[0012] In the above scheme, step (2) is as follows: the algae from step (1) are placed in a sealed bag and stored at 4°C in the dark for 3 to 6 days. Then, they are soaked in disinfected seawater at 90 to 900 times the mass of the algae. The gametes are released after standing for 28 to 69 minutes under the conditions of temperature 26.5±0.1°C to 27.3±0.2°C, humidity 60.4±1.3% to 63.4±0.1%, and light intensity 2500±50Lx to 3250±50Lx.
[0013] In the above scheme, the gamete emission in step (2) specifically refers to: gametes being emitted synchronously in the mature algal region; the gametes are actively swimming, phototactic, and have normal activity; the gamete emission amount is (2.10±0.20)×10 11 ~(2.22±0.03)×10 11 1 / g algae.
[0014] In the above scheme, step (2) is as follows: the algae from step (1) are placed in a sealed bag and stored at 4°C in the dark for 9 to 15 days. Then, they are soaked in disinfected seawater at 90 to 900 times the mass of the algae and cultured at 18°C, light intensity of 3500±50Lx to 4000±50Lx, and photoperiod of 12L:12D for 23 to 50 hours before gametes are released.
[0015] In the above scheme, the gamete emission in step (2) specifically refers to: gametes being emitted synchronously in the mature algal region; the gametes are actively swimming, phototactic, and have normal activity; the gamete emission amount is (2.08±0.36)×10 11 ~(2.28±0.58)×10 11 1 / g algae.
[0016] In the above scheme, the air drying in step (1) is as follows: the algae are air-dried for 5 hours under the following conditions: temperature 24.9±0℃~28±0℃, humidity 39.3±0.3%~63.4±0.1%, light intensity 74.3±7.0Lx~124.6±9.6Lx, so that the surface of the algae is wrinkled and the water content is 37.61±1.36%~56.16±5.14%.
[0017] In the above scheme, the salinity of the disinfected seawater is 25.7 to 30.9‰, and it is obtained by filtration through a sieve cloth with a pore size of 48μm, sterilization in a high-pressure steam sterilizer, and cooling to room temperature.
[0018] In the above scheme, the mass of the algae in step (2) is the mass of the algae after it has been air-dried and sealed and stored at 4°C in the dark.
[0019] Compared with the prior art, the present invention air-dries the mature gametophytes of the reef membrane, causing their surface to wrinkle and lose a certain amount of water, and then seals and preserves them under low temperature and dark conditions at 4°C. This achieves the purpose of not damaging the gametophytes of the reef membrane, delaying the release of gametophytes by 3 to 15 days, and ensuring that the amount of gametophytes released is not significantly different from that before preservation at low temperature and dark conditions at 4°C, thus meeting the needs of production practice. Detailed Implementation
[0020] To make the objectives, technical solutions, and advantages of this invention clearer, the invention will be further described in detail below with reference to embodiments. It should be understood that the specific embodiments described herein are merely illustrative and not intended to limit the invention.
[0021] This invention provides a method for delaying the release of gametes from reef gametophytes, the method comprising:
[0022] Step (1): At room temperature, place the collected, robust, mature, and unreleased reef gametophytes on a clean workbench for cleaning and air drying;
[0023] Specifically, the mature gametophyte refers to an algal body whose top is dark yellow or brownish-yellow, with the yellow area occupying 1 / 3 to 2 / 3 of the entire algal body.
[0024] Specifically, cleaning the algae means unfolding the algae, gently brushing the upper and lower surfaces of the algae several times with a clean, dry soft-bristled brush, and then gently brushing the immature parts of the middle and lower parts of the upper and lower surfaces of the algae several times with a moist soft-bristled brush dipped in disinfected seawater to remove dirt, including mud, sand and large miscellaneous algae, from the upper and lower surfaces of the algae.
[0025] Specifically, the air-drying process involves air-drying the algae for 5 hours at a temperature of 24.9±0℃~28±0℃, a humidity of 39.3±0.3%~63.4±0.1%, and a light intensity of 74.3±7.0Lx~124.6±9.6Lx, resulting in a wrinkled surface and a water content of 37.61±1.36%~56.16±5.14%.
[0026] Step (2): Place the algae from step (1) in a sealed bag and store it at 4°C in the dark for 3 to 6 days. Then soak it in 90 to 900 times the mass of the algae in disinfected seawater. After standing for 28 to 69 minutes under the conditions of temperature 26.5±0.1°C to 27.3±0.2°C, humidity 60.4±1.3% to 63.4±0.1%, and light intensity 2500±50Lx to 3250±50Lx, release the gametes.
[0027] Specifically, gametes are released synchronously in the maturation region of the thallus. These gametes are actively swimming, phototactic, and exhibit normal viability; the gamete release rate is (2.10±0.20)×10 11 ~(2.22±0.03)×10 11 1 / g algae.
[0028] The disinfected seawater has a salinity of 25.7–30.9‰ and is obtained by filtration through a sieve with a pore size of 48 μm, followed by sterilization in a high-pressure steam sterilizer and cooling to room temperature.
[0029] The thallus mass refers to the mass of the thallus after it has been air-dried and sealed and stored at 4°C in the dark for 3–6 days.
[0030] This invention also provides a method for delaying the release of gametes from reef gametophytes, the method comprising:
[0031] Step (1): At room temperature, place the collected, robust, mature, and unreleased reef gametophytes on a clean workbench for cleaning and air drying;
[0032] Specifically, the mature gametophyte refers to an algal body whose top is dark yellow or brownish-yellow, with the yellow area occupying 1 / 3 to 2 / 3 of the entire algal body.
[0033] Specifically, cleaning the algae means unfolding the algae, gently brushing the upper and lower surfaces of the algae several times with a clean, dry soft-bristled brush, and then gently brushing the immature parts of the middle and lower parts of the upper and lower surfaces of the algae several times with a moist soft-bristled brush dipped in disinfected seawater to remove dirt, including mud, sand and large miscellaneous algae, from the upper and lower surfaces of the algae.
[0034] Specifically, the air-drying process involves air-drying the algae for 5 hours at a temperature of 24.9±0℃~28±0℃, a humidity of 39.3±0.3%~63.4±0.1%, and a light intensity of 74.3±7.0Lx~124.6±9.6Lx, resulting in a wrinkled surface and a water content of 37.61±1.36%~56.16±5.14%.
[0035] Step (2): Place the algae from step (1) in a sealed bag and store it at 4°C in the dark for 9 to 15 days. Then soak it in 90 to 900 times the mass of the algae in disinfected seawater. After static culture for 23 to 50 hours at a temperature of 18°C, a light intensity of 3500±50Lx to 4000±50Lx, and a photoperiod of 12L:12D, the gametes are released.
[0036] Specifically, gametes are released synchronously in the maturation region of the thallus. These gametes are actively swimming, phototactic, and exhibit normal viability; the gamete release rate is (2.08±0.36)×10 11~(2.28±0.58)×10 11 1 / g algae.
[0037] The disinfected seawater has a salinity of 25.7–30.9‰ and is obtained by filtration through a sieve with a pore size of 48 μm, followed by sterilization in a high-pressure steam sterilizer and cooling to room temperature.
[0038] The thallus mass refers to the mass of the thallus after it has been air-dried and sealed and stored at 4°C in the dark for 9–15 days.
[0039] Example 1:
[0040] Preparation of disinfected seawater: Take a certain amount of seawater with a salinity of 30‰, filter it through a sieve cloth with a pore size of 48μm, disinfect it in a high-pressure steam sterilizer, and cool it to room temperature for later use.
[0041] At room temperature, place the collected, robust, mature reef gametophytes that have not yet released gametes on a clean workbench, spread the algae out, and gently brush the upper and lower surfaces of the algae several times with a clean, dry soft-bristled brush. Then, use a moist soft-bristled brush dipped in disinfected seawater to gently brush the immature parts of the middle and lower parts of the upper and lower surfaces of the algae several times to remove dirt, including mud, sand, and large miscellaneous algae, from the upper and lower surfaces of the algae.
[0042] Then, it is dried for 5 hours under the conditions of 25℃, 45% humidity and 85Lx light intensity, so that the surface of the algae is wrinkled and the water content is 40%.
[0043] The dried algae were sealed in an airtight bag and stored at 4°C in darkness for 3 days. Then, they were immersed in 100 times their weight of sterilized seawater and allowed to stand for 38 minutes at 26.8°C, 62% humidity, and 2800 Lx light intensity. During this time, the mature algal regions simultaneously released gametes, which were active, phototactic, and of normal viability. The gamete release rate was (2.22 ± 0.03) × 10⁻⁶. 11 1 / g algae.
[0044] Example 2:
[0045] Preparation of disinfected seawater: Take a certain amount of seawater with a salinity of 29‰, filter it through a sieve with a pore size of 48μm, disinfect it in a high-pressure steam sterilizer, and cool it to room temperature for later use.
[0046] At room temperature, place the collected, robust, mature reef gametophytes that have not yet released gametes on a clean workbench, spread the algae out, and gently brush the upper and lower surfaces of the algae several times with a clean, dry soft-bristled brush. Then, use a moist soft-bristled brush dipped in disinfected seawater to gently brush the immature parts of the middle and lower parts of the upper and lower surfaces of the algae several times to remove dirt, including mud, sand, and large miscellaneous algae, from the upper and lower surfaces of the algae.
[0047] Then, it is dried for 5 hours under the conditions of 26℃, 48% humidity and 100Lx light intensity, so that the surface of the algae is wrinkled and the water content is 45%.
[0048] The dried algae were sealed in an airtight bag and stored at 4°C in darkness for 6 days. Then, they were immersed in 600 times their weight of sterilized seawater and allowed to stand for 65 minutes at 26.8°C, 62.5% humidity, and 3000 Lx light intensity. During this time, the mature algal regions simultaneously released gametes, which were active, phototactic, and of normal viability. The gamete release rate was (2.10 ± 0.20) × 10⁻⁶. 11 1 / g algae.
[0049] Example 3:
[0050] Preparation of disinfected seawater: Take a certain amount of seawater with a salinity of 28‰, filter it through a sieve cloth with a pore size of 48μm, disinfect it in a high-pressure steam sterilizer, and cool it to room temperature for later use.
[0051] At room temperature, place the collected, robust, mature reef gametophytes that have not yet released gametes on a clean workbench, spread the algae out, and gently brush the upper and lower surfaces of the algae several times with a clean, dry soft-bristled brush. Then, use a moist soft-bristled brush dipped in disinfected seawater to gently brush the immature parts of the middle and lower parts of the upper and lower surfaces of the algae several times to remove dirt, including mud, sand, and large miscellaneous algae, from the upper and lower surfaces of the algae.
[0052] Then, it was dried for 5 hours at a temperature of 27℃, a humidity of 55%, and a light intensity of 110Lx, so that the surface of the algae was wrinkled and the water content was 52%.
[0053] The dried algae were sealed in airtight bags and stored at 4°C in darkness for 9 days. Then, they were immersed in 900 times their weight of sterilized seawater and allowed to stand for 24 hours at 18°C, with a light intensity of 3500 Lx and a photoperiod of 12 L:12 D. During this time, the mature algal regions synchronously released gametes, which were active, phototactic, and of normal viability. The gamete release rate was (2.08 ± 0.36) × 10⁻⁶. 11 1 / g algae.
[0054] Example 4:
[0055] Preparation of disinfected seawater: Take a certain amount of seawater with a salinity of 27‰, filter it through a sieve cloth with a pore size of 48μm, disinfect it in a high-pressure steam sterilizer, and cool it to room temperature for later use.
[0056] At room temperature, place the collected, robust, mature reef gametophytes that have not yet released gametes on a clean workbench, spread the algae out, and gently brush the upper and lower surfaces of the algae several times with a clean, dry soft-bristled brush. Then, use a moist soft-bristled brush dipped in disinfected seawater to gently brush the immature parts of the middle and lower parts of the upper and lower surfaces of the algae several times to remove dirt, including mud, sand, and large miscellaneous algae, from the upper and lower surfaces of the algae.
[0057] Then, it was dried for 5 hours at a temperature of 26.8℃, a humidity of 53%, and a light intensity of 115Lx, so that the surface of the algae was wrinkled and the water content was 50%.
[0058] The dried algae were sealed in airtight bags and stored at 4°C in darkness for 12 days. They were then immersed in 850 times their mass of sterilized seawater and allowed to stand for 47 hours at 18°C, with a light intensity of 3800 Lx and a photoperiod of 12 L:12 D. The mature algal regions synchronously released gametes, which were active, phototactic, and of normal viability; the gamete release rate was 2.28 × 10⁻⁶. 11 1 / g algae.
[0059] Example 5:
[0060] Preparation of disinfected seawater: Take a certain amount of seawater with a salinity of 27‰, filter it through a sieve cloth with a pore size of 48μm, disinfect it in a high-pressure steam sterilizer, and cool it to room temperature for later use.
[0061] At room temperature, place the collected, robust, mature reef gametophytes that have not yet released gametes on a clean workbench, spread the algae out, and gently brush the upper and lower surfaces of the algae several times with a clean, dry soft-bristled brush. Then, use a moist soft-bristled brush dipped in disinfected seawater to gently brush the immature parts of the middle and lower parts of the upper and lower surfaces of the algae several times to remove dirt, including mud, sand, and large miscellaneous algae, from the upper and lower surfaces of the algae.
[0062] Then, it was dried for 5 hours at a temperature of 26.5℃, a humidity of 50%, and a light intensity of 120Lx, so that the surface of the algae was wrinkled and the water content was 48%.
[0063] The dried algae were sealed in airtight bags and stored at 4°C in darkness for 15 days. Then, they were immersed in 900 times their weight of sterilized seawater and allowed to stand for 50 hours at 18°C, with a light intensity of 4000 Lx and a photoperiod of 12 L:12 D. During this time, the mature algal regions synchronously released gametes, which were active, phototactic, and of normal viability; the gamete release rate was 2.26 × 10⁻⁶. 11 1 / g algae.
[0064] This invention is based on the adaptability of reef gametophytes to environmental factors during reproduction. It utilizes methods such as altering the water content within the algae and low-temperature sealing for preservation to maintain the viability of the gametophytes while delaying gamete release and collection. Depending on the specific circumstances, mature gametophytes can be appropriately dried and stored at 4°C in darkness for an appropriate time to inhibit and delay gamete release. This allows for precise timing of gamete collection, achieving 100% gamete release and obtaining a normal release rate to meet production needs.
[0065] This invention is simple to operate and has the advantages of delaying the collection of reef gametes, effectively shortening the summering time of reef sporophytes, reducing the labor intensity of summering management, and reducing cultivation and production costs.
Claims
1. A method of delaying spawning of a reef fish gamete, characterized by, The method comprises: Step (1): at room temperature, the collected appearance robust, mature, not to discharge the gametophyte of reef membrane gametes is placed on clean bench for cleaning, dry; the step (1) in dry is: under the condition of temperature 24.9±0 ℃-28±0 ℃, humidity 39.3±0.3 %-63.4±0.1 %, illumination intensity 74.3±7.0 Lx-124.6±9.6 Lx, air dry 5h, make the surface of algal body shrink, water content is 37.61±1.36 %-56.16±5.14 %; Step (2): the algal body of step (1) is placed in a dense bag and sealed, stored at 4℃ in the dark, then soaked in 90-900 times the weight of the algal body in sterilized seawater, and then left to stand under the conditions of temperature, humidity and illumination intensity, and then the gametes are discharged.
2. The method of delaying spawning of the reef fish gametes according to claim 1, wherein, The step (2) is: the algal body of step (1) is placed in a dense bag and sealed, stored at 4℃ in the dark for 3-6 days, then soaked in 90-900 times the weight of the algal body in sterilized seawater, and then left to stand under the conditions of temperature 26.5±0.1 ℃-27.3±0.2 ℃, humidity 60.4±1.3 %-63.4±0.1 %, illumination intensity 2500±50 Lx-3250±50 Lx, and then the gametes are discharged after 28-69 minutes.
3. The method of delaying spawning of the reef fish gametes according to claim 2, wherein, In step (2), gamete release specifically involves the synchronous release of gametes in the mature thallus region. These gametes are actively swimming, phototactic, and exhibit normal viability. The gamete release amount is (2.10 ± 0.20) × 10⁻⁶. 11 ~(2.22±0.03)×10 11 1 / g algae.
4. The method of delaying spawning of the reef fish gametes according to claim 1, wherein, The step (2) is: the algal body of step (1) is placed in a dense bag and sealed, stored at 4℃ in the dark for 9-15 days, then soaked in 90-900 times the weight of the algal body in sterilized seawater, and then left to stand under the conditions of temperature 18℃, illumination intensity 3500±50 Lx-4000±50 Lx, and light cycle 12L:12D, and then the gametes are discharged after 23-50h of incubation.
5. The method of delaying spawning of the reef fish gametes according to claim 4, wherein, In step (2), gamete release specifically involves the synchronous release of gametes in the mature thallus region. These gametes are actively swimming, phototactic, and exhibit normal viability. The gamete release amount is (2.08 ± 0.36) × 10⁻⁶. 11 ~(2.28±0.58)×10 11 1 / g algae.
6. The method of delaying spawning of the reef fish gametes according to claim 1, wherein, The salinity of the sterilized seawater is 25.7-30.9 ‰, and the sterilized seawater is filtered through a 48 μm mesh silk cloth and sterilized in a high-pressure steam sterilization pot, and then cooled to room temperature.
7. The method of delaying spawning of the reef fish gametes according to claim 1, wherein, The weight of the algal body in step (2) is the weight of the algal body after being air dried and stored at 4℃ in the dark in a sealed bag.