Method for delaying discharge of monostroma gametes

Abstract

The invention discloses a method for delaying discharge of monostroma gametophytes, which comprises the following steps: at room temperature, cleaning and drying collected monostroma gametophytes which are robust in appearance, mature and not discharged with gametophytes in the shade, sealing the monostroma gametophytes in a zip-lock bag, storing the monostroma gametophytes at 4 DEG C in darkness, soaking the monostroma gametophytes in disinfected seawater which is 90-900 times of the mass of the monostroma gametophytes in the mass of the monostroma gametophytes, and culturing the monostroma gametophytes under the conditions of temperature, humidity and discharging gametes after standing. According to the method, the mature monostroma gametophyte is treated, the discharge time of the monostroma gametophyte is delayed by 3-15 days compared with normal discharge time, the gametophyte can discharge 100% of gametophyte, and the discharge amount of the gametophyte is not obviously different from that of the gametophyte which is dried in the shade, 4 DEG C and treated in the dark. The method is simple and convenient to operate, and has the advantages that the collection of monostroma gametes is delayed, the summer time of monostroma sporophytes is effectively shortened, the labor intensity of summer management is reduced, and the cultivation production cost is reduced.

Description

Technical Field

[0001] The invention belongs to the technical field of reproduction regulation in the process of large-scale economic seaweed cultivation, and in particular relates to a method for delaying the discharge of gametes by reef membrane gametophytes. Background Art

[0002] Monostroma nitidum is a large economic green algae with large gametophytes, rich in nutrients, and high edible and medicinal economic value. When the gametophytes mature, they form gametangia. After the gametes are released, they attach to the substrate and germinate and grow into small sporophytes. When the sporophytes mature, they release spores, which germinate and grow into gametophytes.

[0003] Reef film gametophytes are the algae used for reef film seedling cultivation and planting. In the production practice of reef film cultivation, the emission time, speed and quantity of its gametes are the key factors affecting production, and the emission of gametes is affected by ecological factors such as water temperature, salinity and light intensity. Under suitable conditions, the emission of gametes is concentrated and large in quantity. On the contrary, the emission is dispersed, small in quantity or even non-existent.

[0004] In the practice of reef film seedling cultivation and cultivation, due to adverse weather or human factors, although the reef film gametes are mature, the conditions for collecting gametes are not met and the collection of gametes needs to be delayed. At this time, the mature gametes need to be processed to prevent the gametes from discharging gametes too early, otherwise the gamete discharge will lead to production stagnation. When the adverse factors are removed, the gametes of the reef film need to be collected normally, and whether the treated mature gametes can discharge gametes normally is related to whether the production can proceed smoothly.

[0005] In addition, in production, after collecting reef membrane gametes, they are usually attached to the substrate and cultivated to sporophyte, and the sporophyte is cultivated in summer. Delaying the discharge of reef membrane gametes and thus delaying the collection of reef membrane gametes can effectively shorten the summer cultivation time of reef membrane sporophyte, reduce the labor intensity of its summer management, and reduce the cost of cultivation production.

[0006] At present, there are no reports on the research on the delayed gamete emission from reef membrane gametophytes. Summary of the invention

[0007] In view of this, the main object of the present invention is to provide a method for delaying the discharge of gametes from reef membrane gametophytes.

[0008] To achieve the purpose, the technical solution of the present invention is achieved as follows:

[0009] The present invention provides a method for delaying the discharge of reef membrane gametes, the method comprising:

[0010] Step (1): at room temperature, place the collected reef membrane gametocytes that are robust, mature, and have not released gametes on a clean workbench for cleaning and shade drying;

[0011] Step (2): The algae of step (1) are sealed in a sealed bag and stored at 4°C in the dark. The algae are then immersed in sterilized seawater at a volume of 90 to 900 times the mass of the algae and allowed to stand under the conditions of temperature, humidity and light intensity to allow gametes to be released.

[0012] In the above scheme, step (2) is: place the algae of step (1) in a sealed bag, store it at 4°C in the dark for 3 to 6 days, and then soak it in sterilized seawater 90 to 900 times the mass of the algae, and let it stand 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 before gamete discharge.

[0013] In the above scheme, the gamete discharge in step (2) is specifically as follows: the mature area of ​​the algae discharges gametes synchronously, the gametes swim actively, have phototropism, and have normal vitality; the gamete discharge amount is (2.10±0.20)×10 11 ~(2.22±0.03)×10 11 / g algae.

[0014] In the above scheme, step (2) is: place the algae of step (1) in a sealed bag, store it at 4°C in the dark for 9 to 15 days, then immerse it in sterilized seawater 90 to 900 times the mass of the algae, and culture it at a temperature of 18°C, a light intensity of 3500±50Lx to 4000±50Lx, and a photoperiod of 12L:12D for 23 to 50 hours before gametes are released.

[0015] In the above scheme, the gamete discharge in step (2) is specifically as follows: the mature area of ​​the algae discharges gametes synchronously, the gametes swim actively, have phototropism, and have normal vitality; the gamete discharge amount is (2.08±0.36)×10 11 ~(2.28±0.58)×10 11 / g algae.

[0016] In the above scheme, the shade drying in step (1) is: leaving it in the air for 5 hours under the conditions of temperature 24.9±0℃~28±0℃, humidity 39.3±0.3%~63.4±0.1%, and light intensity 74.3±7.0Lx~124.6±9.6Lx, so that the surface of the algae shrinks and the water content is 37.61±1.36%~56.16±5.14%.

[0017] In the above scheme, the salinity of the sterilized seawater is 25.7-30.9‰, and it is obtained by filtering through a mesh silk cloth with a pore size of 48 μm, sterilizing 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 being shade-dried and sealed and stored at 4° C. in the dark.

[0019] Compared with the prior art, the present invention shade-dries the mature gametes of the reef membrane to make their surface wrinkled and lose water to a certain extent, and then seals and preserves them at a low temperature of 4°C in the dark, so as to achieve the purpose of not damaging the reef membrane gametes, delaying the discharge of reef membrane gametes by 3 to 15 days, and ensuring that the amount of gametes discharged is not significantly different from that before storage at a low temperature of 4°C in the dark, so as to meet the needs of production practice. DETAILED DESCRIPTION

[0020] In order to make the purpose, technical solution and advantages of the present invention more clearly understood, the present invention is further described in detail below in conjunction with the embodiments. It should be understood that the specific embodiments described herein are only used to explain the present invention and are not used to limit the present invention.

[0021] An embodiment of the present invention provides a method for delaying the discharge of gametes by reef membrane gametocytes, the method comprising:

[0022] Step (1): at room temperature, the collected reef membrane gametocytes that are robust, mature, and have not released gametes are placed on a clean workbench for cleaning and shade drying;

[0023] Specifically, the mature gametophyte refers to an algae whose top is dark yellow or brownish yellow, and the yellow area occupies 1 / 3 to 2 / 3 of the area of ​​the entire algae.

[0024] Specifically, the cleaning of the algae refers to spreading the algae, lightly brushing the upper and lower surfaces of the algae with a clean, dry soft-bristle brush for several times, and then lightly brushing the middle and lower immature parts of the upper and lower surfaces of the algae with a moistened soft-bristle brush dipped in sterilized seawater for several times to remove dirt on the upper and lower surfaces of the algae, including mud, sand and large algae.

[0025] Specifically, the shade drying is to expose the algae to the air 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, so that the surface of the algae shrinks and the water content is 37.61±1.36%~56.16±5.14%.

[0026] Step (2): The algae of step (1) are placed in a sealed bag and stored at 4°C in the dark for 3 to 6 days, then immersed in sterilized seawater 90 to 900 times the mass of the algae, and allowed to stand 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, before gamete discharge.

[0027] Specifically, the mature areas of the algae synchronously released gametes, which were active, phototactic, and had normal vitality. The amount of gamete release was (2.10±0.20)×10 11 ~(2.22±0.03)×10 11 / g algae.

[0028] The salinity of the sterilized seawater is 25.7-30.9‰, and the seawater is filtered through a silk cloth with a pore size of 48 μm, sterilized in a high-pressure steam sterilizer, and cooled to room temperature.

[0029] The algae mass refers to the mass of the algae after being shade-dried and sealed and stored at 4° C. in the dark for 3 to 6 days.

[0030] An embodiment of the present invention also provides a method for delaying the discharge of gametes by reef membrane gametocytes, the method comprising:

[0031] Step (1): at room temperature, the collected reef membrane gametocytes that are robust, mature, and have not released gametes are placed on a clean workbench for cleaning and shade drying;

[0032] Specifically, the mature gametophyte refers to an algae whose top is dark yellow or brownish yellow, and the yellow area occupies 1 / 3 to 2 / 3 of the area of ​​the entire algae.

[0033] Specifically, the cleaning of the algae refers to spreading the algae, lightly brushing the upper and lower surfaces of the algae with a clean, dry soft-bristle brush for several times, and then lightly brushing the middle and lower immature parts of the upper and lower surfaces of the algae with a moistened soft-bristle brush dipped in sterilized seawater for several times to remove dirt on the upper and lower surfaces of the algae, including mud, sand and large algae.

[0034] Specifically, the shade drying is to expose the algae to the air 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, so that the surface of the algae shrinks and the water content is 37.61±1.36%~56.16±5.14%.

[0035] Step (2): The algae of step (1) are sealed in a sealed bag and stored at 4°C in the dark for 9 to 15 days, then immersed in sterilized seawater 90 to 900 times the mass of the algae, and cultured at a temperature of 18°C, a light intensity of 3500±50Lx to 4000±50Lx, and a photoperiod of 12L:12D for 23 to 50 hours before gamete discharge.

[0036] Specifically, the mature areas of the algae synchronously released gametes, which were active, phototactic, and had normal vitality. The amount of gamete release was (2.08±0.36)×10 11~(2.28±0.58)×10 11 / g algae.

[0037] The salinity of the sterilized seawater is 25.7-30.9‰, and the seawater is filtered through a silk cloth with a pore size of 48 μm, sterilized in a high-pressure steam sterilizer, and cooled to room temperature.

[0038] The algae mass refers to the mass of the algae after being shade-dried and sealed and stored at 4° C. in the dark for 9 to 15 days.

[0039] Embodiment 1:

[0040] Preparation of sterilized seawater: Take some seawater with a salinity of 30‰, filter it through a silk cloth with a pore size of 48 μm, sterilize it in a high-pressure steam sterilizer, cool it to room temperature, and set it aside.

[0041] At room temperature, place the collected reef membrane gametophytes that are healthy, mature, and have not released gametes on a clean workbench, unfold the algae, and gently brush the upper and lower surfaces of the algae several times with a clean, dry soft-bristle brush. Then use a moistened soft-bristle brush dipped in sterilized 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 on the upper and lower surfaces of the algae, including mud, sand, and large algae.

[0042] Then, it was placed in the air for 5 hours to dry under the conditions of temperature 25°C, humidity 45%, and light intensity 85Lx, so that the surface of the algae was wrinkled and the water content was 40%.

[0043] The shade-dried algae were sealed in a tight bag and stored at 4°C in the dark for 3 days. Then, they were immersed in sterilized seawater 100 times the mass of the algae. After standing for 38 minutes at a temperature of 26.8°C, a humidity of 62%, and a light intensity of 2800Lx, the mature areas of the algae synchronously released gametes. The gametes swam actively, were phototactic, and had normal vitality. The amount of gametes released was (2.22±0.03)×10 11 / g algae.

[0044] Embodiment 2:

[0045] Preparation of sterilized seawater: Take some seawater with a salinity of 29‰, filter it through a silk cloth with a pore size of 48 μm, sterilize it in a high-pressure steam sterilizer, cool it to room temperature, and set it aside.

[0046] At room temperature, place the collected reef membrane gametophytes that are healthy, mature, and have not released gametes on a clean workbench, unfold the algae, and gently brush the upper and lower surfaces of the algae several times with a clean, dry soft-bristle brush. Then use a moistened soft-bristle brush dipped in sterilized 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 on the upper and lower surfaces of the algae, including mud, sand, and large algae.

[0047] Then, it was placed in the air for 5 hours to dry at a temperature of 26°C, a humidity of 48%, and a light intensity of 100 Lx, so that the surface of the algae was wrinkled and the water content was 45%.

[0048] The shade-dried algae were sealed in a tight bag and stored at 4°C in the dark for 6 days. Then, they were immersed in sterilized seawater 600 times the mass of the algae. After standing for 65 minutes at a temperature of 26.8°C, a humidity of 62.5%, and a light intensity of 3000Lx, the mature areas of the algae synchronously released gametes. The gametes swam actively, were phototactic, and had normal vitality. The amount of gametes released was (2.10±0.20)×10 11 / g algae.

[0049] Embodiment 3:

[0050] Preparation of sterilized seawater: Take some seawater with a salinity of 28‰, filter it through a silk cloth with a pore size of 48 μm, sterilize it in a high-pressure steam sterilizer, cool it to room temperature, and set it aside.

[0051] At room temperature, place the collected reef membrane gametophytes that are healthy, mature, and have not released gametes on a clean workbench, unfold the algae, and gently brush the upper and lower surfaces of the algae several times with a clean, dry soft-bristle brush. Then use a moistened soft-bristle brush dipped in sterilized 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 on the upper and lower surfaces of the algae, including mud, sand, and large algae.

[0052] Then, it was left to dry for 5 hours at a temperature of 27°C, a humidity of 55%, and a light intensity of 110 Lx, so that the surface of the algae was wrinkled and the water content was 52%.

[0053] The shade-dried algae were sealed in a tight bag and stored at 4°C in the dark for 9 days. Then, they were immersed in sterilized seawater 900 times the mass of the algae. After standing for 24 hours at 18°C, light intensity 3500Lx, and photoperiod 12L:12D, the mature areas of the algae synchronously released gametes. The gametes swam actively, were phototactic, and had normal vitality. The gamete discharge was (2.08±0.36)×10 11 / g algae.

[0054] Embodiment 4:

[0055] Preparation of sterilized seawater: Take some seawater with a salinity of 27‰, filter it through a silk cloth with a pore size of 48 μm, sterilize it in a high-pressure steam sterilizer, cool it to room temperature, and set it aside.

[0056] At room temperature, place the collected reef membrane gametophytes that are healthy, mature, and have not released gametes on a clean workbench, unfold the algae, and gently brush the upper and lower surfaces of the algae several times with a clean, dry soft-bristle brush. Then use a moistened soft-bristle brush dipped in sterilized 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 on the upper and lower surfaces of the algae, including mud, sand, and large algae.

[0057] Then, it was placed in the air for 5 hours to dry at a temperature of 26.8°C, 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 shade-dried algae were sealed in a tight bag and stored at 4°C in the dark for 12 days. Then, they were immersed in sterilized seawater 850 times the mass of the algae. After standing for 47 hours at 18°C, 3800Lx light intensity, and 12L:12D photoperiod, the mature areas of the algae synchronously released gametes. The gametes swam actively, were phototropic, and had normal vitality. The gamete discharge was 2.28×10 11 / g algae.

[0059] Embodiment 5:

[0060] Preparation of sterilized seawater: Take some seawater with a salinity of 27‰, filter it through a silk cloth with a pore size of 48 μm, sterilize it in a high-pressure steam sterilizer, cool it to room temperature, and set it aside.

[0061] At room temperature, place the collected reef membrane gametophytes that are healthy, mature, and have not released gametes on a clean workbench, unfold the algae, and gently brush the upper and lower surfaces of the algae several times with a clean, dry soft-bristle brush. Then use a moistened soft-bristle brush dipped in sterilized 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 on the upper and lower surfaces of the algae, including mud, sand, and large algae.

[0062] Then, it was placed in the air for 5 hours to dry under the conditions of temperature 26.5℃, humidity 50% and light intensity 120Lx, so that the surface of the algae was wrinkled and the water content was 48%.

[0063] The shade-dried algae were sealed in a tight bag and stored at 4°C in the dark for 15 days. Then, they were immersed in sterilized seawater 900 times the mass of the algae. After standing for 50 hours at 18°C, 4000Lx light intensity, and 12L:12D photoperiod, the mature areas of the algae released gametes synchronously. The gametes swam actively, were phototropic, and had normal vitality. The gamete discharge was 2.26×10 11 / g algae.

[0064] The present invention is based on the adaptation range of the reproduction of reef membrane gametophytes to environmental factors, and changes the water content in the algae and seals and preserves them at low temperature, so as to achieve the purpose of preserving the vitality of the reef membrane gametophytes and delaying the emission of gametes and the collection of gametes. According to the actual situation, the present invention can perform appropriate drying treatment on the mature gametophytes, and store them at a low temperature of 4°C and in the dark for an appropriate time to inhibit the emission of gametes and delay the emission of gametes, so as to grasp the good time of collecting reef membrane gametes, achieve 100% emission of gametes by the gametophytes, and obtain a normal amount of gamete emission to meet the production needs.

[0065] The invention is easy to operate and has the advantages of delaying the collection of reef membrane gametes, effectively shortening the summer time of reef membrane spores, reducing the labor intensity of summer management thereof, and reducing the cultivation production cost.

Claims

1. A method for delaying the discharge of reef membrane gametes, characterized in that: The method includes: Step (1): at room temperature, the collected reef membrane gametocytes that are robust, mature, and have not released gametes are placed on a clean workbench for cleaning and shade drying; Step (2): The algae of step (1) are sealed in a sealed bag and stored at 4°C in the dark. The algae are then immersed in sterilized seawater at a volume of 90 to 900 times the mass of the algae and allowed to stand under the conditions of temperature, humidity and light intensity to allow gametes to be released.

2. The method for delaying the discharge of reef membrane gametes according to claim 1, characterized in that: The step (2) is as follows: the algae of the step (1) are sealed in a sealed bag, stored at 4°C in the dark for 3 to 6 days, and then immersed in sterilized seawater 90 to 900 times the mass of the algae, and placed at a temperature of 26.5±0.1°C to 27.3±0.2°C, a humidity of 60.4±1.3% to 63.4±0.1%, and a light intensity of 2500±50Lx to 3250±50Lx for 28 to 69 minutes before gamete discharge.

3. The method for delaying the discharge of reef membrane gametes according to claim 2, characterized in that: The specific gamete discharge in step (2) is as follows: the mature area of ​​the algae discharges gametes synchronously, the gametes swim actively, have phototropism, and have normal vitality; the gamete discharge amount is (2.10±0.20)×10 11 ~(2.22±0.03)×10 11 / g algae.

4. The method for delaying the discharge of reef membrane gametes according to claim 1, characterized in that: The step (2) is as follows: the algae of the step (1) are sealed in a sealed bag, stored at 4°C in darkness for 9 to 15 days, then immersed in sterilized seawater 90 to 900 times the mass of the algae, and statically cultured 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, before gamete discharge.

5. The method for delaying the discharge of reef membrane gametes according to claim 4, characterized in that: The specific gamete discharge in step (2) is as follows: the mature area of ​​the algae discharges gametes synchronously, the gametes swim actively, have phototropism, and have normal vitality; the gamete discharge amount is (2.08±0.36)×10 11 ~(2.28±0.58)×10 11 / g algae.

6. The method for delaying the discharge of reef membrane gametes according to claim 1, characterized in that: The shade drying in step (1) is: leaving the algae in the air for 5 hours at a temperature of 24.9±0°C to 28±0°C, a humidity of 39.3±0.3% to 63.4±0.1%, and a light intensity of 74.3±7.0Lx to 124.6±9.6Lx, so that the algae surface shrinks and the water content is 37.61±1.36% to 56.16±5.14%.

7. The method for delaying the discharge of reef membrane gametes according to claim 1, characterized in that: The salinity of the sterilized seawater is 25.7-30.9‰, and the seawater is filtered through a silk cloth with a pore size of 48 μm, sterilized in a high-pressure steam sterilizer, and cooled to room temperature.

8. The method for delaying the discharge of reef membrane gametes according to claim 1, characterized in that: The mass of the algae in step (2) is the mass of the algae after being shade-dried and sealed and stored at 4° C. in the dark.