A method for mutation breeding based on Porphyra haitanensis

Abstract

This application belongs to the field of breeding technology and specifically provides a method for mutagenesis breeding of Porphyra haitanensis, comprising the following steps: 1) irradiating the coniferous bodies of Porphyra haitanensis with ultraviolet light; 2) treating the irradiated coniferous bodies with a mutagen comprising nitrosoguanidine and a slow-release solution comprising bleomycin and bleomycin; and 3) screening the mutagenized coniferous bodies to obtain mutants, which are then cultured to obtain thallus. This breeding method has the advantages of high efficiency, high mutation rate, and good specificity.

Description

Technical Field

[0001] The present application belongs to the field of breeding technology, and in particular relates to a method for induction breeding based on Pyropia haitanensis. Background Art

[0002] Porphyra belongs to the taxonomic classes Protoflorideae, Bangiales, and Bangiaceae of the phylum Rhodophyta and is distributed across cold, temperate, subtropical, and tropical waters. Porphyra is highly nutritious, rich in protein, polysaccharides, lipids, vitamins, and minerals. Due to its high polysaccharide and phycoerythrin content and unique biological functions, it has broad application prospects in medicine, food, cosmetics, and animal nutrition.

[0003] Due to the impacts of climate change, fluctuating sea conditions, overharvesting, and grazing by algae-eating fish and shellfish, natural resources are dwindling. To date, large-scale cultivation of Porphyra haitanensis still primarily relies on self-cultivated seeds, resulting in significant degradation of germplasm, reduced yield, and decreased quality, impacting the health and sustainability of the laver industry. Therefore, screening for high-quality, stress-resistant new germplasm of Porphyra haitanensis remains a crucial requirement for industrial development.

[0004] Acquiring genetic variants of laver is a crucial step in laver breeding. Using physical and chemical mutagenesis methods, phycologists have generated a large number of laver color mutants, providing extensive research material for advancing scientific progress in laver genetics, physiology, and other research areas. Artificial color mutants of laver are primarily obtained through physical and chemical mutagenesis. Mutagenesis breeding is the most widely used artificial mutagenesis method. Unlike traditional breeding methods, which suffer from long breeding cycles and unstable traits, it effectively improves breeding efficiency and allows for the targeted development of new varieties based on breeding goals.

[0005] Currently, radiation can induce mutations quickly, but it is easily absorbed by the chromosomes, causing significant damage to the chromosome structure and resulting in relatively random mutation sites. Chemical mutagenesis has a high mutation rate and is site-specific, but it also has a wide mutation range and is less efficient. Therefore, it is necessary to find a highly efficient and superior breeding method for jar cabbage. Summary of the Invention

[0006] In view of the above problems, in order to further improve the effect of induced breeding of Pyrrophyta haitanensis, the present application provides a method for induced breeding of Pyrrophyta haitanensis.

[0007] The present application provides a method for mutagenesis breeding based on Porphyra haitanensis, comprising the following steps:

[0008] 1) UV irradiation of the conchoceles of Porphyra haitanensis;

[0009] 2) treating the filaments after ultraviolet irradiation in step 1) with a mutagen, wherein the mutagen comprises nitrosoguanidine and a slow-release solution, wherein the slow-release solution comprises bleomycin and bleomycin;

[0010] 3) Screen the filaments after mutagenesis to obtain mutant strains, and continue to culture to obtain thallus.

[0011] Furthermore, in step 1), the filaments are statically cultured in MES culture medium before ultraviolet irradiation, wherein the MES culture medium contains 5-15 μg / L copper sulfate.

[0012] Furthermore, in step 1), the irradiation dose of ultraviolet radiation is 20-50 J / m 2 .

[0013] Furthermore, in step 2), the mutagenesis time is 20-60 min.

[0014] Furthermore, in step 2), the concentration of nitrosoguanidine in the mutagen is 15-60 μg / mL.

[0015] Furthermore, in step 2), the concentration of bleomycin in the sustained-release solution is 300-500 ppm.

[0016] Furthermore, in step 2), the concentration of bleomycin in the sustained-release solution is 100-200 ppm.

[0017] Furthermore, in step 2), the sustained-release solution is prepared by a method comprising the following steps:

[0018] S1: Mix cyclodextrin, polyethylene glycol, and deionized water to prepare a base solution;

[0019] S2: placing amino polystyrene particles in domestic sewage, culturing them statically, then washing them with ultrapure water, sterilizing them, and drying them to obtain biofilm@amino polystyrene particles;

[0020] S3: Add biofilm@amino polystyrene particles, bleomycin and bleomycin to the base liquid and mix well.

[0021] Furthermore, in step S2, the average particle size of the amino polystyrene particles is 15-80 μm.

[0022] Furthermore, in step S3, the content of biofilm@amino polystyrene particles in the sustained-release solution is 0.5-0.85 wt%.

[0023] Compared with the prior art, this application has the following beneficial effects:

[0024] 1. This application first subjects the filaments to a certain degree of low-dose ultraviolet irradiation, so that the filamentous cells are fully activated, and then treats them with mutagens, which can increase the mutation efficiency and number of filamentous cells.

[0025] 2. This application introduces a sustained-release system of bleomycin and bleomycin, based on nitrosoguanidine induction, to further enhance mutation efficiency and specificity. Furthermore, the biofilm@aminopolystyrene particles adsorb and control the release of bleomycin and bleomycin, increasing mutant survival. Furthermore, the biofilm@aminopolystyrene particles themselves provide a certain degree of induction, thereby enhancing effective induction and promoting the subsequent formation of conchosporangium branches from mutant filaments. BRIEF DESCRIPTION OF THE DRAWINGS

[0026] Figure 1 Schematic diagram of the development of mutant conidia in the induced breeding method of Porphyra haitanensis according to Examples 1-2 and Control Groups 1-2 of the present application. DETAILED DESCRIPTION

[0027] The following will be combined with the drawings in the embodiments of this application to clearly and completely describe the technical solutions in the embodiments of this application. Obviously, the embodiments described are only part of the embodiments of this application, not all of the embodiments. Based on the embodiments in this application, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of this application.

[0028] After a large number of experimental studies and explorations of various induction breeding methods, this application found that low-dose ultraviolet irradiation treatment can provide a mutation basis for chemical induction, and establish bleomycin and bleomycin auxiliary induction in a slow-release solution environment to further enhance the induction effect.

[0029] Specifically, the present application provides a method for induction breeding of Porphyra haitanensis, comprising the following steps:

[0030] 1) UV irradiation of the conchoceles of Porphyra haitanensis;

[0031] 2) treating the filaments after ultraviolet irradiation in step 1) with a mutagen, wherein the mutagen comprises nitrosoguanidine and a slow-release solution, wherein the slow-release solution comprises bleomycin and bleomycin;

[0032] 3) Screen the filaments after mutagenesis to obtain mutant strains, and continue to culture to obtain thallus.

[0033] Furthermore, in step 1), the filaments are statically cultured in MES culture medium before ultraviolet irradiation, wherein the MES culture medium contains 5-15 μg / L copper sulfate.

[0034] In some specific embodiments, in step 1), the filaments are statically cultured in MES culture medium before UV irradiation. The MES culture medium may contain 5 μg / L, 6 μg / L, 7 μg / L, 8 μg / L, 9 μg / L, 10 μg / L, 11 μg / L, 12 μg / L, 13 μg / L, 14 μg / L, or 15 μg / L of copper sulfate. More preferably, in step 1), when the MES culture medium contains 10 μg / L of copper sulfate, better experimental results are generally achieved.

[0035] Furthermore, in step 1), the irradiation dose of ultraviolet radiation is 20-50 J / m 2 .

[0036] In some embodiments, in step 1), the irradiation dose of ultraviolet radiation can be 20 J / m 2 , 25J / m 2 、30J / m 2 、35J / m 2 40J / m 2 45J / m 2 , 50J / m 2 More preferably, under normal circumstances, in step 1), the irradiation dose of ultraviolet radiation is 30J / m 2 When , better experimental results can be obtained.

[0037] Furthermore, in step 2), the mutagenesis time is 20-60 min.

[0038] In some specific embodiments, in step 2), the mutagenesis time can be 20 min, 25 min, 30 min, 35 min, 40 min, 45 min, 50 min, 55 min, or 60 min. More preferably, under normal circumstances, in step 2), a mutagenesis time of 25 min can achieve better experimental results.

[0039] Furthermore, in step 2), the concentration of nitrosoguanidine in the mutagen is 15-60 μg / mL.

[0040] In some specific embodiments, in step 2), the concentration of nitrosoguanidine in the mutagen can be 15 μg / mL, 20 μg / mL, 25 μg / mL, 30 μg / mL, 35 μg / mL, 40 μg / mL, 45 μg / mL, 50 μg / mL, 55 μg / mL, or 60 μg / mL. More preferably, under normal circumstances, in step 2), a concentration of nitrosoguanidine in the mutagen of 20 μg / mL can achieve better experimental results.

[0041] Furthermore, in step 2), the concentration of bleomycin in the sustained-release solution is 300-500 ppm.

[0042] In some specific embodiments, in step 2), the concentration of bleomycin in the sustained-release solution may be 300 ppm, 310 ppm, 320 ppm, 330 ppm, 340 ppm, 350 ppm, 360 ppm, 370 ppm, 380 ppm, 390 ppm, 400 ppm, 410 ppm, 420 ppm, 430 ppm, 440 ppm, 450 ppm, 460 ppm, 470 ppm, 480 ppm, 490 ppm, or 500 ppm. More preferably, under normal circumstances, in step 2), when the concentration of bleomycin in the sustained-release solution is 380 ppm, better experimental results can be achieved.

[0043] Furthermore, in step 2), the concentration of bleomycin in the sustained-release solution is 100-200 ppm.

[0044] In some specific embodiments, in step 2), the concentration of bleomycin in the sustained-release solution can be 100 ppm, 110 ppm, 120 ppm, 130 ppm, 140 ppm, 150 ppm, 160 ppm, 170 ppm, 180 ppm, 190 ppm, or 200 ppm. Generally, in step 2), a bleomycin concentration of 120 ppm in the sustained-release solution achieves optimal experimental results.

[0045] Furthermore, in step 2), the sustained-release solution is prepared by a method comprising the following steps:

[0046] S1: Mix cyclodextrin, polyethylene glycol, and deionized water to prepare a base solution;

[0047] S2: placing amino polystyrene particles in domestic sewage, culturing them statically, then washing them with ultrapure water, sterilizing them, and drying them to obtain biofilm@amino polystyrene particles;

[0048] S3: Add biofilm@amino polystyrene particles, bleomycin and bleomycin to the base liquid and mix well.

[0049] Furthermore, in step S2, the average particle size of the amino polystyrene particles is 15-80 μm.

[0050] In some specific embodiments, in step S2, the average particle size of the amino polystyrene particles may be 15 μm, 20 μm, 25 μm, 30 μm, 35 μm, 40 μm, 45 μm, 50 μm, 55 μm, 60 μm, 65 μm, 70 μm, 75 μm, or 80 μm. More preferably, under normal circumstances, in step S2, when the average particle size of the amino polystyrene particles is 60 μm, better experimental results can be obtained.

[0051] Furthermore, in step S3, the content of biofilm@amino polystyrene particles in the sustained-release solution is 0.5-0.85 wt%.

[0052] In some specific embodiments, in step S3, the content of the biofilm@aminopolystyrene particles in the sustained-release solution may be 0.5wt%, 0.55wt%, 0.6wt%, 0.65wt%, 0.7wt%, 0.75wt%, 0.8wt%, or 0.85wt%. More preferably, under normal circumstances, in step S3, when the content of the biofilm@aminopolystyrene particles in the sustained-release solution is 0.8wt%, better experimental results can be achieved.

[0053] Example 1

[0054] MES culture medium preparation: accurately weigh sodium nitrate 1.4g, sodium glycerophosphate 0.2g, tris (hydroxymethyl)aminomethane 2g, EDTA-Na2 0.3g, boric acid 0.25g, MnCl 2· 4H2O0.035g, FeCl3.6H2O12.5mg, ZnCl22.5mg, CoCl2.6H2O1mg, and ferric citrate 60mg were added to a sterile cup and the volume was adjusted to 500mL with ultrapure water to obtain MES culture medium mother solution. 20mL of MES culture medium mother solution was added to 1000mL of seawater (specific gravity 1.022), and then copper sulfate was added and mixed thoroughly to obtain the MES culture medium. The copper sulfate content in the MES culture medium was controlled to be 10μg / L.

[0055] Inducer preparation: Add nitrosoguanidine and 20 mL of sustained-release solution to 1000 mL of sterile seawater (specific gravity 1.022) and bring to volume. Maintain a nitrosoguanidine concentration of 20 μg / mL in the inducer.

[0056] The sustained-release solution is prepared by a method comprising the following steps:

[0057] S1: Prepare the base solution by mixing 3 g of cyclodextrin, 15 mL of polyethylene glycol, and 200 mL of deionized water.

[0058] S2: Add bleomycin and bleomycin to the base solution and mix well. Control the concentration of bleomycin in the sustained-release solution to be 380 ppm; the concentration of bleomycin in the sustained-release solution to be 120 ppm.

[0059] The mutagenesis breeding method based on Porphyra haitanensis of the present embodiment comprises the following steps:

[0060] 1) Wild Porphyra haitanensis (NH, collected from Pingtan County, Fujian Province) was stored in the laboratory for future use. The storage conditions were: MES medium, temperature 23 ± 1 °C, light intensity 4 μmol photons / (m 2 s), photoperiod 12L:12D, half of the MES medium was replaced every 180 days;

[0061] The filaments were chopped into pieces with a grinder to a length of about 100 μm, and then inoculated into a culture dish and cultured for 10 days. Then, a short-wave ultraviolet lamp was used as the irradiation source for ultraviolet irradiation, and the irradiation dose was controlled at 20-50 J / m 2 ;

[0062] 2) Add 5 mL of mutagen to the culture dish and treat the filaments after UV irradiation in step 1) with the mutagen for 25 minutes. Then, wash the culture dish with MES medium to remove residual mutagen and replenish MES medium. Incubate at 20°C, 2500 Lux, and a photoperiod of 12 L:12 D, replacing the medium every 3 days.

[0063] 3) Cut the induced filaments into small pieces using a tissue crusher, select and place them in a 48-well plate, and culture them in the dark at 20°C for 3 days. Then gradually transfer them to normal light with 50 μmol photons / (m 2 s). After one month, the number of surviving conifers was counted and the surviving conifers were transferred to cell culture flasks. Samples with more and larger thallus were selected as experimental materials for screening to obtain mutants, which were further cultured to obtain thallus.

[0064] Example 2

[0065] MES culture medium preparation: accurately weigh 1.4 g sodium nitrate, 0.2 g sodium glycerophosphate, 2 g tris (hydroxymethyl)aminomethane, 0.3 g EDTA-Na2, 0.25 g boric acid, and MnCl 2·4H2O0.035g, FeCl3.6H2O12.5mg, ZnCl22.5mg, CoCl2.6H2O1mg, and ferric citrate 60mg were added to a sterile cup and the volume was adjusted to 500mL with ultrapure water to obtain MES culture medium mother solution. 20mL of MES culture medium mother solution was added to 1000mL of seawater (specific gravity 1.022), and then copper sulfate was added and mixed thoroughly to obtain the MES culture medium. The copper sulfate content in the MES culture medium was controlled to be 10μg / L.

[0066] Inducer preparation: Add nitrosoguanidine and 20 mL of sustained-release solution to 1000 mL of sterile seawater (specific gravity 1.022) and bring to volume. Maintain a nitrosoguanidine concentration of 20 μg / mL in the inducer.

[0067] The sustained-release solution is prepared by a method comprising the following steps:

[0068] S1: Prepare the base solution by mixing 3 g of cyclodextrin, 15 mL of polyethylene glycol, and 200 mL of deionized water.

[0069] S2: 20 g of amino polystyrene particles (average particle size of amino polystyrene particles is 60 μm) were placed in 500 mL of domestic sewage (pH 7.31, COD Cr 98.25mg / L, TN9.82mg / L, NH4 + -N8.58mg / L), stirred at 150rpm and cultured at 25℃ for 30 days. After the culture was completed, the particles were filtered, washed with ultrapure water, sterilized, and dried to obtain biofilm@amino polystyrene particles.

[0070] S3: Add biofilm@aminopolystyrene particles, bleomycin and bleomycin to the base liquid and mix well. Control the concentration of bleomycin in the sustained-release solution to be 380 ppm; the concentration of bleomycin in the sustained-release solution to be 120 ppm; and the content of biofilm@aminopolystyrene particles in the sustained-release solution to be 0.8 wt%.

[0071] The mutagenesis breeding method based on Porphyra haitanensis of the present embodiment comprises the following steps:

[0072] 1) Wild Porphyra haitanensis (NH, collected from Pingtan County, Fujian Province) was stored in the laboratory for future use. The storage conditions were: MES medium, temperature 23 ± 1 °C, light intensity 4 μmol photons / (m 2 s), photoperiod 12L:12D, half of the MES medium was replaced every 180 days;

[0073] The filaments were chopped into pieces with a grinder to a length of about 100 μm, and then inoculated into a culture dish and cultured for 10 days. Then, a short-wave ultraviolet lamp was used as the irradiation source for ultraviolet irradiation, and the irradiation dose was controlled at 20-50 J / m 2 ;

[0074] 2) Add 5 mL of mutagen to the culture dish and treat the filaments after UV irradiation in step 1) with the mutagen for 25 minutes. Then, wash the culture dish with MES medium to remove residual mutagen and replenish MES medium. Incubate at 20°C, 2500 Lux, and a photoperiod of 12 L:12 D, replacing the medium every 3 days.

[0075] 3) Cut the induced filaments into small pieces using a tissue crusher, select and place them in a 48-well plate, and culture them in the dark at 20°C for 3 days. Then gradually transfer them to normal light with 50 μmol photons / (m 2 s). After one month, the number of surviving conifers was counted and the surviving conifers were transferred to cell culture flasks. Samples with more and larger thallus were selected as experimental materials for screening to obtain mutants, which were further cultured to obtain thallus.

[0076] Control group 1

[0077] MES culture medium preparation: accurately weigh 1.4 g sodium nitrate, 0.2 g sodium glycerophosphate, 2 g tris (hydroxymethyl)aminomethane, 0.3 g EDTA-Na2, 0.25 g boric acid, and MnCl 2· 4H2O0.035g, FeCl3.6H2O12.5mg, ZnCl22.5mg, CoCl2.6H2O1mg, and ferric citrate 60mg were added to a sterile cup and the volume was adjusted to 500mL with ultrapure water to obtain MES culture medium mother solution. 20mL of MES culture medium mother solution was added to 1000mL of seawater (specific gravity 1.022), and then copper sulfate was added and mixed thoroughly to obtain the MES culture medium. The copper sulfate content in the MES culture medium was controlled to be 10μg / L.

[0078] Inducer preparation: Add nitrosoguanidine to 1000 mL of sterile seawater (specific gravity 1.022) and bring to volume. Maintain a nitrosoguanidine concentration of 20 μg / mL.

[0079] The control group's mutagenesis breeding method based on Porphyra haitanensis includes the following steps:

[0080] 1) Wild Porphyra haitanensis (NH, collected from Pingtan County, Fujian Province) was stored in the laboratory for future use. The storage conditions were: MES medium, temperature 23 ± 1 °C, light intensity 4 μmol photons / (m 2s), photoperiod 12L:12D, half of the MES medium was replaced every 180 days;

[0081] The filaments were chopped into pieces with a grinder to a length of about 100 μm, and then inoculated into a culture dish and cultured for 10 days. Then, a short-wave ultraviolet lamp was used as the irradiation source for ultraviolet irradiation, and the irradiation dose was controlled at 20-50 J / m 2 ;

[0082] 2) Add 5 mL of mutagen to the culture dish and treat the filaments after UV irradiation in step 1) with the mutagen for 25 minutes. Then, wash the culture dish with MES medium to remove residual mutagen and replenish MES medium. Incubate at 20°C, 2500 Lux, and a photoperiod of 12 L:12 D, replacing the medium every 3 days.

[0083] 3) Cut the induced filaments into small pieces using a tissue crusher, select and place them in a 48-well plate, and culture them in the dark at 20°C for 3 days. Then gradually transfer them to normal light with 50 μmol photons / (m 2 s). After one month, the number of surviving conifers was counted and the surviving conifers were transferred to cell culture flasks. Samples with more and larger thallus were selected as experimental materials for screening to obtain mutants, which were further cultured to obtain thallus.

[0084] Control group 2

[0085] MES culture medium preparation: accurately weigh sodium nitrate 1.4g, sodium glycerophosphate 0.2g, tris (hydroxymethyl)aminomethane 2g, EDTA-Na2 0.3g, boric acid 0.25g, MnCl 2· 4H2O0.035g, FeCl3.6H2O12.5mg, ZnCl22.5mg, CoCl2.6H2O1mg, and ferric citrate 60mg were added to a sterile cup and the volume was adjusted to 500mL with ultrapure water to obtain MES culture medium mother solution. 20mL of MES culture medium mother solution was added to 1000mL of seawater (specific gravity 1.022), and then copper sulfate was added and mixed thoroughly to obtain the MES culture medium. The copper sulfate content in the MES culture medium was controlled to be 10μg / L.

[0086] Inducer preparation: Add nitrosoguanidine to 1000 mL of sterile seawater (specific gravity 1.022) and bring to volume. Maintain a nitrosoguanidine concentration of 20 μg / mL.

[0087] The control group's mutagenesis breeding method based on Porphyra haitanensis includes the following steps:

[0088] 1) Wild Porphyra haitanensis (NH, collected from Pingtan County, Fujian Province) was stored in the laboratory for future use. The storage conditions were: MES medium, temperature 23 ± 1 °C, light intensity 4 μmol photons / (m 2 s), photoperiod 12L:12D, half of the MES medium was replaced every 180 days;

[0089] 2) Use a grinder to chop the filaments into pieces of approximately 100 μm in length, then inoculate them into culture dishes and culture them for 10 days. Then, add 5 mL of mutagen to the culture dish and treat with the mutagen for 25 minutes. Then, wash the culture dish with MES culture medium to remove residual mutagen, and then replenish MES culture medium. Incubate at 20°C, 2500 Lux, and a photoperiod of 12 L:12 D, replacing the culture medium every 3 days.

[0090] 3) Cut the induced filaments into small pieces using a tissue crusher, select and place them in a 48-well plate, and culture them in the dark at 20°C for 3 days. Then gradually transfer them to normal light with 50 μmol photons / (m 2 s). After one month, the number of surviving conifers was counted and the surviving conifers were transferred to cell culture flasks. Samples with more and larger thallus were selected as experimental materials for screening to obtain mutants, which were further cultured to obtain thallus.

[0091] Performance testing

[0092] 1. According to the mutagenesis breeding method based on Porphyra haitanensis of Examples 1-2 and Control Groups 1-2, the mutation rate and mortality rate of the conifers were counted, the mutation rate = S1 / S2, S1 is the number of conifers at the beginning of the experiment, and S2 is the number of conifers that mutated at the end of the experiment; the mortality rate = S3 / S4, S3 is the number of conifers at the beginning of the experiment, and S4 is the number of conifers that died at the end of the experiment; the results are shown in Table 1. It can be seen that the breeding method of the present application has a more efficient and specific induction effect, and stable mutants can be obtained.

[0093] Table 1 Mutagenesis data of conchoderma in the mutagenesis breeding method based on Porphyra haitanensis in Examples 1-2 and Control Groups 1-2 of the present application

[0094] Serial number mortality rate / % Mutation rate / % Example 1 49.36 15.37 Example 2 50.58 18.12 Control group 1 38.64 10.93 Control group 2 20.15 7.11

[0095] 2. The filaments of the mutant strains of Example 1-2 and Control 1-2 were crushed, and 0.07 g (fresh weight) was taken and cultured at 20°C for 6 weeks. The development of the filaments was observed under a microscope. The results were as follows: Figure 1As shown, a is Example 1, b is Example 2, c is Control group 1, and d is Control group 2. It can be seen that the mutant filaments obtained by the breeding method of the present application have better growth and development status and are more likely to form conchosporon branches than the control group.

[0096] Although the present application has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments, or make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present application should be included in the scope of protection of the present invention.

Claims

1. A method for induction breeding of Porphyra haitanensis, characterized in that: The steps include: 1) The filaments of Porphyra haitanensis were irradiated with ultraviolet radiation at a dose of 20-50 J / m 2 ; 2) treating the filaments after UV irradiation in step 1) with a mutagen for 20-60 minutes; the mutagen comprises nitrosoguanidine and a sustained-release solution, the sustained-release solution comprises bleomycin and bleomycin; the concentration of nitrosoguanidine in the mutagen is 15-60 μg / mL; the concentration of bleomycin in the sustained-release solution is 300-500 ppm, and the concentration of bleomycin in the sustained-release solution is 100-200 ppm; 3) Screen the filaments after mutagenesis to obtain mutant strains, and continue to culture to obtain thallus.

2. The method for induced breeding of Porphyra haitanensis according to claim 1, wherein: In the step 1), the filaments are statically cultured in MES culture medium before being irradiated with ultraviolet light, wherein the MES culture medium contains 5-15 μg / L of copper sulfate.

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