Method for efficiently obtaining porphyra yezoensis fruitspore seedlings
By fragmenting laver with a wall-breaking machine and culturing it under specific aeration conditions, combined with shell culture, the problems of low efficiency and seasonal limitations in carpospore collection have been solved, achieving efficient acquisition of carpospore seedlings.
Patent Information
- Application Number
- CN202511636203.4
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2026-01-30
- Publication Date
- 2026-02-27
AI Technical Summary
In existing technologies, the collection efficiency of Porphyra stripes fruit spores is low and subject to seasonal limitations, resulting in a cumbersome seedling collection process and making it difficult to achieve rapid and efficient acquisition of fruit spore seedlings.
The laver is fragmented using a high-speed blender, collected by sieve, and cultured under specific conditions with aeration. The fragments are then mixed with seashells for further culture, which promotes the release of carpospores in large quantities and their penetration into the shells within a few days.
It enables rapid acquisition of caryopsis seedlings, breaks seasonal limitations, improves seedling collection efficiency, and shortens the time for caryopsis formation.
Smart Images
Figure CN121569740A_ABST
Abstract
Description
TECHNICAL FIELD
[0001] The present application relates to the sexual reproduction and breeding technology of Porphyra, and particularly relates to a method for efficiently obtaining fruiting spore seedlings of Porphyra yezoensis. BACKGROUND
[0002] The life history of Porphyra yezoensis presents the phenomenon of alternate generations, including the sporophyte generation and the gametophyte generation. The sporophyte of Porphyra yezoensis is diploid, which is filamentous, small in size, branched, and grows in the shell in nature; the gametophyte generation of Porphyra yezoensis is haploid, which is manifested as the leaf-shaped body for sea area cultivation and the Porphyra sold in the market.
[0003] Compared with free filamentous bodies, the shell filamentous bodies cultivated by the fruiting spore seedling method have the advantages of low technical threshold and simple equipment requirement, and the mature shell spores are easier to disperse. Therefore, the shell filamentous body seedling of Porphyra yezoensis generally adopts this method. The process is as follows: first, high-quality Porphyra is selected as seed Porphyra, fruiting spores are collected, the fruiting spores are uniformly sprayed on the washed and classified clam shells and other shell substrates after being diluted with water, and the fruiting spores will germinate and drill into the shells to form filamentous bodies under suitable conditions.
[0004] At present, the bottleneck of fruiting spore collection of Porphyra yezoensis is the low efficiency of fruiting spore collection. First, the amount of fruiting spores released at a time is small: after the leaf-shaped body of Porphyra yezoensis matures, sperm sacs and carpospheres will form on its surface, and the combination of the two forms fruiting spore sacs, which in turn produce fruiting spores. This process is gradually completed, and the amount of fruiting spores released at a time is not much. Second, the collection period of fruiting spores is long and is limited by seasons, and the seed Porphyra must be mature and release fruiting spores in a specific season to carry out seedling collection. Therefore, how to realize the rapid acquisition of fruiting spore seedlings and break through the seasonal limitation is of great significance to the sustainable development of Porphyra yezoensis cultivation industry. SUMMARY
[0005] The present application provides a method for efficiently obtaining fruiting spore seedlings of Porphyra yezoensis to make up for the shortcomings of the prior art.
[0006] Technical scheme: A method for efficiently obtaining fruiting spore seedlings of Porphyra yezoensis, comprising the following steps: (1) Preparation of Porphyra fragments: freeze-thawed or fresh Porphyra is crushed by a cell disruptor to obtain Porphyra fragments; (2) Collection of fragments: the Porphyra fragments are filtered by a filter cloth, the filtrate is discarded, and the Porphyra fragments are collected; (3) Fragment culture: the collected Porphyra fragments are aerated and cultured for several days to make the fragment cells develop fruiting spores; (4) Addition of shells: after the fragment cells produce fruiting spores, shells are added and mixed for several days to obtain fruiting spore seedlings.
[0007] The method for efficiently obtaining Porphyra fruiting spore seedlings, in step (1), the Porphyra is crushed by a cell wall breaking machine for 1-3 minutes, and ice is added every 0.3-0.7 minutes during the crushing to prevent the water from overheating.
[0008] The method for efficiently obtaining Porphyra fruiting spore seedlings, in step (1), the Porphyra is crushed by a cell wall breaking machine for 1-3 minutes, and ice is added every 0.3-0.7 minutes during the crushing to prevent the water from overheating.
[0009] The method for efficiently obtaining Porphyra fruiting spore seedlings, in step (2), the screen silk used for collecting the Porphyra fragments is 200-500 meshes, and the collected fragments are washed with clean seawater.
[0010] The method for efficiently obtaining Porphyra fruiting spore seedlings, in step (3), the light intensity for culturing the fragments is 2000-4000 lux, the temperature is 5-15 ℃, and the light cycle is light:dark=10-15 h:10-15 h.
[0011] The method for efficiently obtaining Porphyra fruiting spore seedlings, in step (4), after the fragments produce fruiting spores, shells are added for mixed culturing for several days to obtain fruiting spore seedlings with the fruiting spores drilled into the shells.
[0012] The method for efficiently obtaining Porphyra fruiting spore seedlings, in step (3), the fragments are aerated for 5-7 days.
[0013] The method for efficiently obtaining Porphyra fruiting spore seedlings, in step (4), the mixed culturing is performed for 7-12 days.
[0014] The method for efficiently obtaining Porphyra fruiting spore seedlings, the Porphyra is selected from Porphyra yezoensis.
[0015] The method for efficiently obtaining Porphyra fruiting spore seedlings, the aeration is air.
[0016] The method for efficiently obtaining Porphyra fruiting spore seedlings, the power of the cell wall breaking machine is 2000-2500 W.
[0017] Preferably, in (2), the fragments are collected by using a 500-mesh screen silk, and the fragments are washed with sterile seawater for at least 3 times.
[0018] Preferably, in (3), the culturing conditions of the fragments are as follows: the culture solution is PES, the culturing temperature is 10 ℃, the light intensity is 3000 lux, and the fragments are aerated.
[0019] Preferably, in (4), the fragments are co-cultured with the shells for 7-12 days, and the culturing conditions are as follows: the culture solution is PES, the culturing temperature is 10 ℃, the light intensity is 3000 lux, and the fragments are aerated or static cultured.
[0020] Beneficial effects: Compared with the prior art, the present invention has the following advantages: (1) Compared with the traditional method of collecting fruit spores from mature seaweed and spraying them onto seashells for seedling collection, the present invention directly cultivates mechanically crushed seaweed for several days and then co-cultivates it with seashells, so that the fruit spores are concentrated and released in large quantities within a few days and penetrate the shell, thus shortening the time for seaweed to form fruit spores. (2) Compared with the traditional method of collecting seedlings in a specific season, the present invention can use frozen striped seaweed to recover and collect seedlings, breaking the seasonal restrictions on seaweed shell spore seedling collection. (3) Compared with the traditional method of waiting for seaweed to fully mature, the seaweed used in the present invention is not limited to mature seaweed, but can also be used to collect fruit spore seedlings after crushing immature seaweed. Attached Figure Description
[0021] Figure 1 The spore penetration of shells was observed in mature laver group 2 after 7 days of adding shells.
[0022] Figure 2 The spore penetration of shells after adding shells to mature laver group 1 for 12 days;
[0023] Figure 3 The spore penetration of shells was observed in mature laver group 2 after 12 days of adding shells.
[0024] Figure 4 The spore penetration of shells was observed in mature laver group 3 after 12 days of adding shells.
[0025] Figure 5 The spore penetration of shells after 7 days of adding immature laver fragments to shells;
[0026] Figure 6 The spore penetration of shells after adding immature laver fragments to shells for 12 days;
[0027] Figure 7 The spore penetration of shells was observed in mature laver group 4 after 12 days of adding shells.
[0028] Figure 8 The spore penetration of immature laver complete thallus after 12 days of adding shells. Detailed Implementation
[0029] The technical solution of the present invention will be further described and illustrated below with reference to the embodiments.
[0030] The blender is a Kormes WK-05, 2200 W. It uses a 35 W air pump and is equipped with a 6-hole distributor.
[0031] Unless otherwise specified, all materials and reagents used in the following examples are commercially available.
[0032] Example 1 Disintegration and culture steps: (1) Take the frozen frond of Porphyra yezoensis (mature) with an age of about 72 days to recover. The recovery method is as follows: first, put the sterilized natural seawater into a 4℃ refrigerator and stand for one day; then, transfer the seawater to a 10℃ incubator, and put the frozen Porphyra yezoensis to be recovered into the incubator, and stand in the dark for one day; on the third day, start aeration culture, with a light intensity of 3000 lux and a light cycle of 12 h / 12 h (day / night); (2) Divide the recovered Porphyra yezoensis into three groups, and put them into a broken wall machine containing 500 mL of sterilized seawater to break for 2 minutes. Add ice every half minute during the breaking to prevent overheating damage to the Porphyra yezoensis; (3) Collect the broken pieces of each group using a 500-mesh screen gauze, and rinse them several times (2-6 times) with clean seawater. Weigh 0.2 g of the broken pieces of Porphyra yezoensis for each group; (4) Put the weighed broken pieces of group 1 and group 2 into aeration bottles containing culture solution for aeration culture, and put the weighed broken pieces of group 3 into aeration bottles containing culture solution for static culture, so that they produce carpospores; (5) When the culture reaches the 7th day, the broken pieces start to release carpospores. At this time, add sterilized shell pieces with a thickness of about 1 mm, 4 pieces for each group. After adding the shell pieces, group 1 continues aeration culture, and groups 2 and 3 are static culture, so that the released carpospores drill into the shell; (6) When the culture reaches the 12th day, observe the carpospore drilling under a microscope.
[0033] The culture conditions of steps (4)-(5) are as follows: the culture solution is sterilized seawater added with PES culture medium, the temperature is 10℃, the light intensity is 3000 lux, and the light cycle is 12 h / 12 h (day / night).
[0034] Experimental results: 1. Microscopic observation of carpospore drilling of Porphyra yezoensis In order to explore whether aeration culture can accelerate the time of releasing carpospores of mechanically broken Porphyra yezoensis and whether aeration affects the carpospore drilling of Porphyra yezoensis, the present application sets three different culture conditions for the Porphyra yezoensis (group 1: aeration culture of Porphyra yezoensis fragments, aeration culture after adding shell; group 2: aeration culture of Porphyra yezoensis fragments, static culture after adding shell; group 3: static culture of Porphyra yezoensis fragments, static culture after adding shell); Compared with other groups, on the 7th day after adding the shell, group 2 found that a large number of carpospores germinated into filamentous bodies and drilled into the shell (as shown in Figure 1 ); on the 12th day, it was found that carpospores germinated into filamentous bodies and drilled into the shell in the four groups, among which the carpospore seedlings of group 2 drilled into the shell the most and were the densest (as shown in Figure 3 ); the carpospore seedlings of group 1 grew the fastest and the filamentous bodies were evenly distributed (as shown in Figure 2 ); the carpospore seedlings of group 3 were less than those of group 2, and grew slower than those of group 2 (as shown in Figure 4).
[0035] Example 2 Crushing culture step: (1) Take fresh thallus of Porphyra yezoensis (immature) with age of 45 days, put it into a cell disruptor containing 500 mL sterilized seawater, crush for 2 minutes, and add ice every half minute to prevent overheating and damage to the Porphyra; (2) Collect the crushed pieces using a 500-mesh silk screen, and rinse several times (2-6 times) with clean seawater; (3) Put the weighed crushed pieces into a gas culture bottle containing culture solution, and culture under aeration; (4) When the culture reaches the 7th day, the crushed pieces start to release tetraspores, at this time, add 4 pieces of sterilized shell with a thickness of about 1 mm, and after adding the shell, stand for culture, so that the released tetraspores drill into the shell; (5) When the culture reaches the 12th day, observe the drilling of tetraspores under a microscope.
[0036] The culture conditions of steps (3)-(4) are as follows: the culture solution is sterilized seawater added with PES medium, the temperature is 10°C, the light intensity is 3000 lux, and the light cycle is 12 h / 12 h (day / night).
[0037] Experimental results: immature Porphyra can also be observed to drill into the shell by this method after crushing, Figure 5 the drilling of tetraspore seedlings cultured with the crushed pieces and the shell for 7 days, Figure 6 and the drilling of tetraspore seedlings cultured with the crushed pieces and the shell for 12 days.
[0038] Comparative Example 1 A method similar to Example 1, except that in step (1), the Porphyra is not crushed, and the whole Porphyra is cultured under aeration, and after adding the shell, standing for culture (Group 4).
[0039] Experimental results: 1. Microscopic observation of the drilling of tetraspores of Porphyra yezoensis At the 12th day of culture, only a small amount of tetraspore seedlings were found to drill into the shell (such as Figure 7 ).
[0040] Comparative Example 2
[0041] A method similar to Example 2, except that in step (1), the Porphyra is not crushed, and the whole Porphyra is cultured under aeration, and after adding the shell, standing for culture.
[0042] Experimental results: 1. Microscopic observation of the drilling of tetraspores of Porphyra yezoensis At the 12th day of culture, no boring of the conchocelis by the gametophyte was found (e.g. Figure 8 ).
[0043] After comprehensive analysis of the results, it was found that the broken culture of Examples 1 and 2 significantly increased the number of boring of the conchocelis by the gametophyte compared with the intact culture of Comparative Examples 1 and 2. Therefore, the method for efficiently obtaining the gametophyte of P. yezoensis is as follows: after crushing the P. yezoensis thallus for 2 minutes by the cell wall breaker, the fragments are collected and washed several times, then the fragments are subjected to aeration culture, and after 7 days of aeration culture, the conchae are added and allowed to stand or subjected to aeration culture for several days, and abundant gametophytes (i.e. conchocelis filaments) can be obtained.
Claims
1. A method for efficiently obtaining *Porphyra tenuifolia* spore seedlings, characterized in that, Includes the following steps: (1) Preparation of seaweed fragments: Frozen or fresh seaweed is crushed using a high-speed blender to obtain seaweed fragments; (2) Collection of fragments: Use a sieve to filter the seaweed fragments, discard the filtrate, and collect the seaweed fragments; (3) Fragment culture: The collected seaweed fragments are cultured under aeration for several days to allow the fragment cells to develop into carpospores; (4) Add shells: After the fragmented cells produce carbospores, add shells and mix and culture for several days to obtain carbospore seedlings.
2. The method for efficiently obtaining *Porphyra yezoensis* spore seedlings as described in claim 1, characterized in that, In step (1), use a high-speed blender to crush the seaweed for 1-3 minutes. Add ice every 0.3-0.7 minutes during the crushing process to prevent the water temperature from getting too hot.
3. The method for efficiently obtaining *Porphyra yezoensis* spore seedlings as described in claim 1, characterized in that, Step (1) Select the laver leaves.
4. The method for efficiently obtaining *Porphyra yezoensis* spore seedlings as described in claim 1, characterized in that, Step (2) The sieve used for collecting seaweed fragments is 200-500 mesh. The collected fragments are rinsed with clean seawater.
5. The method for efficiently obtaining *Porphyra yezoensis* spore seedlings as described in claim 1, characterized in that, Step (3) The culture of the fragments is carried out under a light of 2000-4000 lux and a temperature of 5-15 ℃. The photoperiod is light:dark = 10-15 h: 10-15 h.
6. The method for efficiently obtaining *Porphyra yezoensis* spore seedlings as described in claim 1, characterized in that, Step (4) involves adding shells after the fragmented cells produce carpospores, mixing and culturing for several days to obtain carpospore seedlings with carpospores penetrating the shells.
7. The method for efficiently obtaining *Porphyra yezoensis* spore seedlings as described in claim 1, characterized in that, The aerated culture period for the seaweed fragments in step (3) is 5-7 days.
8. The method for efficiently obtaining *Porphyra yezoensis* spore seedlings as described in claim 1 or 6, characterized in that, The mixed culture period in step (4) is 7-12 days.
9. The method for efficiently obtaining *Porphyra yezoensis* spore seedlings as described in claim 1, characterized in that, The laver is selected from Laveria bassiana.
10. The method for efficiently obtaining *Porphyra yezoensis* spore seedlings as described in claim 1, characterized in that, The ventilation refers to the passage of air.