A method for rapidly breeding long-stem grape fern seedlings

Abstract

The invention provides a method for rapidly breeding long-stem grape fern algae seedlings, comprising: (1) taking long-stem grape fern algae as parent algae and cultivating them to maturity; (2) cultivating them in a transition period when male and female gametes are released; (3) transferring them to a new cultivation pond, increasing the light intensity, allowing the parent algae to continue to release male and female gametes, and taking out the parent algae; (4) injecting nutrient water every day within one week of fertilized egg planting; after one week of planting, replacing 10-15% of the volume of pool water with a combination of nutrient water and filtered seawater every 3-4 days, increasing the light intensity of LED green light with a short-term irradiation of 1000-1500Lx every 1-2 days, continuously irradiating for 3-5 days, and irradiating for 2-4 hours every day; until the seedlings are harvested, the invention realizes efficient and stable artificial breeding of long-stem grape fern algae seedlings, shortens the seedling cultivation time to within 30 days, has a high breeding output rate, and the average output within 30 days can reach 10,000 plants / m 2 above.

Description

Technical Field

[0001] The invention relates to the technical field of seaweed cultivation, and in particular to a method for rapidly breeding long-stem cauline fern seedlings. Background Art

[0002] Caulerpa lentillifera, also known as sea grapes, is a large marine green algae that is widely distributed in tropical and subtropical waters. It is rich in polysaccharides, fiber, amino acids, protein and various trace elements, and has attracted much attention due to its high economic and ecological value.

[0003] With the continuous development of seaweed cultivation industry, the existing research on the breeding and propagation technology of long-stem grape fern algae has been continuously deepened. my country has achieved a certain scale of artificial breeding of long-stem grape fern algae in Fujian, Hainan, Qingdao and other regions. Due to the influence of environmental growth factors such as water temperature, salinity, and light during the growth and development of long-stem grape fern algae, the breeding requirements of long-stem grape fern algae are relatively high. The breeding of high-quality and high-yield long-stem grape fern algae seedlings is the basis for the artificial breeding of long-stem grape fern algae, but there are few studies on the breeding and cultivation technology of long-stem grape fern algae seedlings. In the process of cultivating long-stem grape fern algae seedlings, there are often problems such as slow expansion and cultivation of excellent seedlings, unstable yield, and low stress resistance, resulting in low seedling quality. Therefore, it is urgent to find a method for rapid breeding of long-stem grape fern algae seedlings to achieve efficient and stable artificial breeding of long-stem grape fern algae seedlings and promote the more extensive promotion and application of artificial breeding technology of long-stem grape fern algae. Summary of the invention

[0004] In view of this, the present invention proposes a method for rapidly breeding long-stem grape fern seedlings.

[0005] The technical solution of the present invention is achieved in this way:

[0006] The present invention provides a method for rapidly breeding long-stem grape fern algae seedlings, comprising the following steps:

[0007] Step 1: Take a long-stem grape fern algae with an upright stem length of ≥8 cm as the parent algae, place it in a cultivation pool with an attachment base, and carry out mature cultivation;

[0008] Step 2: When the parent algae begin to release male and female gametes, reduce the light intensity to 1200-1600Lx, photoperiod 10L:14D, water temperature 19-23℃, and carry out transition period cultivation for 2-3 days;

[0009] Step 3: Transfer the parent algae cultivated in the transition period to a new cultivation pond, increase the light intensity to 6000-8000Lx, the photoperiod is 12L:12D, the water temperature is 27-30℃, the parent algae continue to release male and female gametes, and then take out the parent algae;

[0010] Step 4: Fertilized egg cultivation: Within one week of fertilized egg implantation, cultivate the eggs under the conditions of light intensity of 1000-2000Lx and photoperiod of 8L:16D, and inject 0.2-0.3kg / m into the cultivation pool every day. 2 Nutrient water is injected 2-3 times a day; one week after the fertilized eggs are settled, 10-15% of the volume of the pool water is replaced with a combination of nutrient water and filtered seawater every 3-4 days, and under the conditions of white light intensity of 2000-3000Lx and photoperiod of 14L:10D, the light intensity of LED green light is increased to 1000-1500Lx for short-term irradiation every 1-2 days, and the irradiation is continuous for 3-5 days, 2-4 hours a day; the fertilized eggs are cultivated under the condition of maintaining the water salinity at 35-37;

[0011] Step 5: When the seedlings grow to more than 4.5cm, they can be harvested.

[0012] It is further explained that the conditions for the transition period cultivation are white light intensity of 1500Lx, photoperiod of 10L:14D, and water temperature of 20-22°C.

[0013] Further explanation, in step 4, the nutrient water is sterilized pool water added with 2-4 mg / L citric acid, 3-6 mg / L ammonium nitrate, 4-8 mg / L sodium chloride, 8-10 mg / L ammonium chloride, 0.5-0.6 mg / L indolebutyric acid, and 0.01-0.05 mg / L biotin.

[0014] Further, in step 4, when the nutrient water body and the filtered seawater are combined to replace the pool water, the volume ratio of the nutrient water body to the filtered seawater is: (1-2): (3-7).

[0015] To further illustrate, in step 4, when 10-15% of the volume of the pool water is replaced with a combination of nutrient water and filtered seawater, the water flow is controlled to be ≤0.2 m / s.

[0016] To further explain, in step 4, under the conditions of white light intensity of 2500Lx and light cycle of 14L:10D, short-term irradiation with LED green light of 1500Lx is added every 2 days, and irradiation is continued for 5 days, 3 hours per day.

[0017] Further, the conditions for the mature cultivation are: water temperature 27-30°C, white light intensity 3000-4000Lx, photoperiod 12L:12D, and the cultivation density of the parent algae is 1.2-1.5g / m 2 .

[0018] It is further explained that the attachment base is made of gravel with a particle size of 0.3-0.4 mm or vinylon net rope laid on the bottom of the pool.

[0019] Compared with the prior art, the present invention has the following beneficial effects:

[0020] (1) After the parent algae are cultivated to sexual maturity, the present invention facilitates increasing the amount of male and female gametes released by the parent algae by cultivating and acclimatizing them during a transition period under low light intensity and low temperature conditions, thereby achieving high yield. In addition, the present invention uses the male and female gametes released after the transition period cultivation for rapid reproduction, and is conducive to enhancing the tolerance of the seedlings to changes in the cultivation water temperature.

[0021] (2) The present invention optimizes the fertilized egg cultivation conditions, utilizes nutrient water and filtered seawater to perform different water fertilization in the fertilized egg colonization stage and the early growth stage, and at the same time, on the basis of regulating the cultivation water environment, uses a certain intensity of LED green light for short-term cycle irradiation to promote the physiological response of the rapid development and growth of the long-stem grape fern algae seedlings, greatly accelerating the breeding rate of the long-stem grape fern algae seedlings, shortening the seedling cultivation time to within 30 days, and achieving a high breeding output rate. The average yield within 30 days can reach 10,000 plants / m 2 At the same time, the light tolerance of the long-stem grape fern algae seedlings is enhanced, which is beneficial to improving the uniformity of upright stem growth and realizing large-scale high-yield and high-quality artificial rapid breeding of long-stem grape fern algae seedlings. DETAILED DESCRIPTION

[0022] In order to better understand the technical content of the present invention, specific embodiments are provided below to further illustrate the present invention.

[0023] Unless otherwise specified, the experimental methods used in the embodiments of the present invention are all conventional methods.

[0024] Unless otherwise specified, the materials, reagents, etc. used in the embodiments of the present invention can be obtained from commercial sources.

[0025] Example 1 - A method for breeding long-stem grape fern algae seedlings, comprising the following steps:

[0026] (1) taking a long-stem grape fern algae with an upright stem length of 8 to 9 cm as a parent algae, placing it in a cultivation pond with an attachment substrate, controlling the water temperature to 27-29°C, the white light intensity to 3000Lx, and the photoperiod to 12L:12D, and carrying out mature cultivation;

[0027] (2) When the parent algae begin to release male and female gametes, reduce the white light intensity to 1200 Lx, the photoperiod is 10 L:14 D, and the water temperature is 19-21 ° C, and the transition period is cultivated for 3 days;

[0028] (3) Transfer the parent algae cultivated in the transition period to a new cultivation pond, increase the light intensity to 6000Lx, the photoperiod to 12L:12D, the water temperature to 27-29°C, the parent algae continue to release male and female gametes, and remove the parent algae;

[0029] (4) Fertilized egg cultivation: Within one week of fertilized egg implantation, the fertilized eggs were cultivated under the conditions of a light intensity of 1000 Lx and a photoperiod of 8 L:16 D, and 0.2 kg / m was injected into the cultivation pool every day. 2 The nutrient water is injected twice a day; the nutrient water is sterilized pool water with 2mg / L citric acid, 3mg / L ammonium nitrate, 4mg / L sodium chloride, 8mg / L ammonium chloride, 0.5mg / L indolebutyric acid, and 0.01mg / L biotin added;

[0030] One week after the fertilized eggs were planted, 10% of the volume of the pool water was replaced every 3 days with a combination of nutrient water and filtered seawater, and the volume ratio of nutrient water to filtered seawater was 1:3; the water flow was controlled at 0.1-0.2 m / s; and under the conditions of white light intensity of 2000Lx and photoperiod of 14L:10D, the light intensity of LED green light (445-465nm) was increased by 1000Lx every 1 day for short-term irradiation, and irradiation was continued for 5 days, 4 hours a day; the fertilized eggs were cultivated under the condition of maintaining the water salinity at 35-36.

[0031] Example 2 - A method for breeding long-stem grape fern algae seedlings, comprising the following steps:

[0032] (1) taking a long-stem grape fern algae with an upright stem length of 8 to 9 cm as a parent algae, placing it in a cultivation pond with an attachment substrate, controlling the water temperature to 28-30°C, the white light intensity to 4000Lx, and the photoperiod to 12L:12D, and carrying out mature cultivation;

[0033] (2) When the parent algae begin to release male and female gametes, reduce the white light intensity to 1600 Lx, the photoperiod is 10 L:14 D, and the water temperature is 20-23 ° C, and the transition period is cultivated for 2 days;

[0034] (3) Transfer the parent algae cultivated in the transition period to a new cultivation pond, increase the light intensity to 8000Lx, the photoperiod to 12L:12D, the water temperature to 28-30°C, the parent algae continue to release male and female gametes, and then remove the parent algae;

[0035] (4) Fertilized egg cultivation: Within one week of fertilized egg implantation, the fertilized eggs were cultivated under the conditions of a light intensity of 2000 Lx and a photoperiod of 8 L:16 D, and 0.3 kg / m was injected into the cultivation pool every day. 2 The nutrient water is injected three times a day; the nutrient water is sterilized pool water with 4mg / L citric acid, 6mg / L ammonium nitrate, 8mg / L sodium chloride, 10mg / L ammonium chloride, 0.6mg / L indolebutyric acid, and 0.05mg / L biotin added;

[0036] One week after the fertilized eggs were planted, 15% of the volume of the pool water was replaced every 4 days with a combination of nutrient water and filtered seawater, and the volume ratio of nutrient water to filtered seawater was 2:7; the water flow was controlled at 0.1-0.2 m / s; and under the conditions of white light intensity of 3000Lx and photoperiod of 14L:10D, the light intensity of LED green light (445-465nm) was increased to 1500Lx for short-term irradiation every 2 days, and irradiation was continued for 3 days, 2 hours per day; the fertilized eggs were cultivated under the condition of maintaining the water salinity at 36-37.

[0037] Example 3 - A method for breeding long-stem grape fern algae seedlings, comprising the following steps:

[0038] (1) taking a long-stem grape fern algae with an upright stem length of 8 to 9 cm as a parent algae, placing it in a cultivation pond with an attachment substrate, controlling the water temperature to 27-29°C, the white light intensity to 3500Lx, and the photoperiod to 12L:12D, and carrying out mature cultivation;

[0039] (2) When the parent algae begin to release male and female gametes, reduce the white light intensity to 1500 Lx, the photoperiod is 10 L:14 D, and the water temperature is 20-22 ° C, and the transition period is cultivated for 2 days;

[0040] (3) Transfer the parent algae cultivated in the transition period to a new cultivation pond, increase the light intensity to 7000Lx, the photoperiod to 12L:12D, the water temperature to 27-29°C, the parent algae continue to release male and female gametes, and then remove the parent algae;

[0041] (4) Fertilized egg cultivation: Within one week of fertilized egg implantation, the fertilized eggs were cultivated under the conditions of a light intensity of 1500 Lx and a photoperiod of 8 L:16 D, and 0.25 kg / m was injected into the cultivation pool every day. 2 The nutrient water is injected three times a day; the nutrient water is sterilized pool water with 3mg / L citric acid, 5mg / L ammonium nitrate, 6mg / L sodium chloride, 9mg / L ammonium chloride, 0.55mg / L indolebutyric acid, and 0.03mg / L biotin added;

[0042] One week after the fertilized eggs were planted, 13% of the volume of the pool water was replaced every 3 days with a combination of nutrient water and filtered seawater, and the volume ratio of nutrient water to filtered seawater was 1.5:6; the water flow was controlled at 0.1-0.2 m / s; and under the conditions of white light intensity of 2500Lx and photoperiod of 14L:10D, the light intensity of LED green light (445-465nm) was increased to 1500Lx every 2 days for short-term irradiation, and irradiation was continued for 5 days, 3 hours a day; the fertilized eggs were cultivated under the condition of maintaining the water salinity at 35-36.

[0043] Comparative Example 1

[0044] As in the method for breeding long-stemmed grape fern algae seedlings of Example 3, the transitional period cultivation of the parent algae is eliminated. In step 1, the parent algae that have been cultivated to maturity are transferred to a new cultivation pond, and directly cultivated with an illumination intensity of 7000Lx, a photoperiod of 12L:12D, and a water temperature of 27-29°C, male and female gametes are released, the parent algae are taken out, and fertilized eggs are cultivated under the same conditions as in Example 3.

[0045] Comparative Example 2

[0046] As in the method for breeding long-stem grape fern algae seedlings of Example 3, during the entire cultivation process of the fertilized eggs, 0.25 kg / m 2 The nutrient water body is injected three times a day; 13% of the volume of the pool water is replaced with filtered seawater every 3 days, and the water flow is controlled at 0.1-0.2 m / s.

[0047] Comparative Example 3

[0048] As in the breeding method of long-stem grape fern seedlings of Example 3, one week after the fertilized eggs were planted, the white light intensity was controlled to 4000Lx, the photoperiod was 14L:10D, and LED green light was not used for short-term irradiation treatment; the other breeding conditions were the same as those in Example 3.

[0049] Comparative Example 4

[0050] As in the breeding method of the long-stem grape fern algae seedlings of Example 3, one week after the fertilized eggs are planted, a combination of LED green light with a light intensity of 1500Lx and white light with a light intensity of 2500Lx is used for irradiation, and the photoperiod is 14L:10D; the other breeding conditions are the same as those in Example 3.

[0051] Example 4 - Effects of different propagation methods of long-stem grape fern algae seedlings on the propagation rate of long-stem grape fern algae seedlings.

[0052] According to the breeding methods of the long-stem grape fern algae seedlings of the above-mentioned Example 3 and Comparative Examples 1-4, a breeding pond containing a pond bottom paved with gravel having a particle size of 0.3-0.4 mm was selected to carry out a long-stem grape fern algae seedling breeding experiment, and the long-stem grape fern algae with an upright stem length of 8-9 cm in the same batch were taken as parent algae, and the breeding density of the parent algae was 1.3 g / m 2 The inlet and outlet water conditions of the cultivation pool were the same, and the growth and development of the parent algae of the long-stem grape fern algae were regularly observed (sexual maturity was the formation of a mesh structure on the surface of the long-stem grape fern algae, and the base of the spherical branchlets grew bud tubes); and the average length of the upright stems on the 20th and 30th days after the fertilized eggs were planted and grown, and the long-stem grape fern algae seedling breeding yield after 30 days was counted. The results are as follows:

[0053]

[0054] As can be seen from the above table, the long-stem grape fern seedling breeding method of Example 3 of the present invention has a high seedling yield, and the breeding growth rate is significantly improved. The upright stems grow in a uniform length, and the seedling cultivation time can be shortened to within 30 days (reaching more than 4.5 cm). The average yield within 30 days can reach 10,200 plants / m 2 .

[0055] By comparing the growth rates of the long-stem grape fern algae seedlings of different experimental groups, it can be seen that the growth rates of the long-stem grape fern algae seedlings of Examples 2, 3, and 4 are reduced. Among them, in Example 2, the water body is fertilized by supplementing nutrient water alone, and the yield of its seedlings is reduced and the growth rate of the seedlings is affected. In Comparative Example 3, short-term cycle irradiation treatment with LED green light of a certain intensity was not used, and it was impossible to effectively obtain high-quality long-stem grape fern seedlings within 30 days, and the average length of its upright stems was less than 4.5 cm. Although LED green light combined with white light was used in Comparative Example 4, under the longer green light irradiation, although the seedlings could be effectively harvested within 30 days (more than 4.5 cm), the tolerance of the seedlings to growth light was low, and the uniformity of the length of the upright stems was poor, indicating that the use of LED green light of a certain intensity for short-term cycle irradiation promotes the physiological response of the rapid development and growth of the long-stem grape fern seedlings, which is beneficial to accelerate the breeding rate of the long-stem grape fern seedlings and enhance the light tolerance of the long-stem grape fern seedlings, and maintain good uniformity of upright stem growth.

[0056] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims

1. A method for rapidly breeding long-stem grape fern algae seedlings, characterized in that: The steps include: Step 1: Take a long-stem grape fern algae with an upright stem length of ≥8 cm as the parent algae, place it in a cultivation pool with an attachment base, and carry out mature cultivation; Step 2: When the parent algae begin to release male and female gametes, reduce the light intensity to 1200-1600Lx, photoperiod 10L:14D, water temperature 19-23℃, and carry out transition period cultivation for 2-3 days; Step 3: Transfer the parent algae cultivated in the transition period to a new cultivation pond, increase the light intensity to 6000-8000Lx, the photoperiod is 12L:12D, the water temperature is 27-30℃, the parent algae continue to release male and female gametes, and then take out the parent algae; Step 4: Fertilized egg cultivation: Within one week of fertilized egg implantation, cultivate the eggs under the conditions of light intensity of 1000-2000Lx and photoperiod of 8L:16D, and inject 0.2-0.3kg / m into the cultivation pool every day. 2 Nutrient water is injected 2-3 times a day; one week after the fertilized eggs are settled, 10-15% of the volume of the pool water is replaced with a combination of nutrient water and filtered seawater every 3-4 days, and under the conditions of white light intensity of 2000-3000Lx and photoperiod of 14L:10D, the light intensity of LED green light is increased to 1000-1500Lx for short-term irradiation every 1-2 days, and the irradiation is continuous for 3-5 days, 2-4 hours a day; the fertilized eggs are cultivated under the condition of maintaining the water salinity at 35-37; Step 5: When the seedlings grow to more than 4.5cm, they can be harvested.

2. A method for rapidly breeding long-stem grape fern algae seedlings as claimed in claim 1, characterized in that: The conditions for the transition period cultivation are white light intensity of 1500Lx, photoperiod of 10L:14D, and water temperature of 20-22°C.

3. A method for rapidly breeding long-stem grape fern algae seedlings as claimed in claim 1, characterized in that: In step 4, the nutrient water body is sterilized pool water added with 2-4 mg / L citric acid, 3-6 mg / L ammonium nitrate, 4-8 mg / L sodium chloride, 8-10 mg / L ammonium chloride, 0.5-0.6 mg / L indolebutyric acid, and 0.01-0.05 mg / L biotin.

4. A method for rapidly breeding long-stem grape fern algae seedlings as claimed in claim 3, characterized in that: In step 4, when the nutrient water body and filtered seawater are combined to replace the pool water, the volume ratio of the nutrient water body to the filtered seawater is: (1-2): (3-7).

5. A method for rapidly breeding long-stem grape fern algae seedlings as claimed in claim 4, characterized in that: In step 4, when replacing 10-15% of the volume of the pool water with a combination of nutrient water and filtered seawater, the water flow is controlled to be ≤0.2 m / s.

6. A method for rapidly breeding long-stem grape fern algae seedlings as claimed in claim 1, characterized in that: In step 4, under the conditions of white light intensity of 2500Lx and photoperiod of 14L:10D, short-term irradiation with LED green light of 1500Lx is added every 2 days for continuous irradiation for 5 days, 3 hours per day.

7. A method for rapidly breeding long-stem grape fern seedlings as claimed in claim 1, characterized in that: The conditions for the mature cultivation are: water temperature 27-30°C, white light intensity 3000-4000Lx, photoperiod 12L:12D, and the cultivation density of the parent algae is 1.2-1.5g / m 2 .

8. The method for rapidly breeding long-stem grape fern algae seedlings as claimed in claim 1, characterized in that: The attachment base is made of gravel with a particle size of 0.3-0.4 mm or vinylon net rope laid on the bottom of the pool.

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