Method for efficiently cultivating gold thread fern by using marchantia polymorpha

Abstract

The application provides a method for efficiently cultivating goldthread fern by using caterpillar waterweed, and relates to the technical field of fern breeding. The method for cultivating goldthread fern mainly comprises the following steps: soaking the caterpillar waterweed in a nutrient solution composed of modified Hoagland's solution and extract of rock rose, and then squeezing the caterpillar waterweed to obtain a substrate; and spreading pure spores of the goldthread fern on the substrate, and spraying clean water and a nutrient agent composed of modified Hoagland's solution and extract of rock rose at different stages to promote the growth of the goldthread fern. The method overcomes the shortcomings of the prior art, effectively improves the germination rate and seedling rate of the goldthread fern spores, reduces the pollution rate, and to a certain extent, shortens the seedling time of the goldthread fern, while ensuring the survival rate of the transplanted seedlings.

Description

Technical Field

[0001] The present invention relates to the technical field of fern propagation, and specifically relates to a method for efficiently cultivating Cibotium barometz using caterpillar sphagnum moss. Background Art

[0002] Cibotium barometz (L.) J. Sm., also known as cibotium rhizome, is a large terrestrial fern belonging to the genus Cibotium in the family Dicksoniaceae. The plant can reach a height of 1-3 meters, and the parts exposed above the ground are densely covered with golden yellow long hairs, shaped like a golden-haired dog, hence the name. Cibotium barometz is the only original plant of "Rhizoma Cibotii", a traditional Chinese medicinal material with a long history in China, which has an application history of more than 1900 years. It has the effects of tonifying the liver and kidney, strengthening the waist and knees, and dispelling wind-dampness, and is one of the main raw materials for more than 100 Chinese patent medicines such as Zhuangyao Jianshen Pills and Shujin Jianyao Pills. Due to over-excavation and destruction of its suitable habitat, the wild resources of Cibotium barometz have sharply decreased. It has been included in the List of National Key Protected Wild Plants (No. 15 of 2021), belonging to the national second-class key protected wild plant, and is also a species listed in Appendix II of the Convention on International Trade in Endangered Species of Wild Fauna and Flora.

[0003] At present, the artificial propagation of Cibotium barometz mainly relies on ramet propagation and spore tissue culture technology. Ramet propagation is limited by the number of original mother plants, with a low propagation coefficient, and cannot meet the demand of large-scale production. Although spore tissue culture propagation can greatly increase the propagation coefficient, the prior art still has the following shortcomings: (1) The tissue culture operation is complicated, requiring multiple subculture transfers under sterile conditions, which imposes high requirements on equipment and technical personnel, leading to high production costs; (2) Tissue culture seedlings are easily affected by environmental factors during the transplanting and domestication process, resulting in unstable seedling formation rates. Direct sowing of spores under natural conditions can avoid the defects of the above two methods, but direct sowing for seedling cultivation mostly uses peat soil, leaf mold, native habitat soil, etc. as seedling substrates, which has problems such as easy compaction of the substrate, poor water retention and air permeability, easy growth of moss and miscellaneous ferns, and easy breeding of molds, resulting in a generally low seedling formation rate. Therefore, selecting a suitable substrate is the key to improving the seedling formation rate of Cibotium barometz. Summary of the Invention

[0004] In view of the deficiencies in the prior art, the present invention provides a method for efficiently cultivating Cibotium barometz using caterpillar sphagnum moss. The method is simple to operate, low in cost, can significantly improve the spore germination rate and seedling formation rate of Cibotium barometz, and is suitable for large-scale popularization and application.

[0005] To achieve the above purpose, the present invention adopts the following technical solution: A method for efficiently cultivating Cibotium barometz using caterpillar sphagnum moss, said method comprising the following steps: S1. Collect pure spores of Cibotium barometz for later use; S2. After diluting the modified Hoglund nutrient solution to 1 / 10-1 / 2, add rock sage extract to adjust the content to 7-14 g / L to obtain the nutrient solution. The rock sage extract is obtained by extracting rock sage with organic solvent by heating and reflux, filtering, and concentrating and drying the filtrate. S3. After sterilizing the caterpillar sphagnum moss, soak it in the above nutrient solution for 2-4 hours, then take it out and squeeze it dry until it no longer drips, and then spread it evenly in the seedling container to obtain the caterpillar sphagnum moss substrate for later use. S4. Evenly spread the pure spores of *Cibotium barometz* onto the surface of the *Sphagnum moss* substrate. Spray with sterile water to ensure close adhesion between the spores and the *Sphagnum moss* surface. Then cover the substrate and place it in an environment with a temperature of 25±2℃, relative humidity of 70%-80%, and shading rate of 70%-80% for routine management until the spores germinate and form sporophyte seedlings. The specific methods for routine management include the following steps: S4-1. From sowing until the gametophytes are visible to the naked eye, spray pure water every 3-5 days to keep the caterpillar sphagnum moss substrate moist but not waterlogged. S4-2. From the time the gametophyte becomes visible to the naked eye until the sporophyte seedlings are formed, spray nutrient solution 1, which consists of 1 / 4-1 / 2 modified Hoglund and 5-7 g / L of rock sage extract, every 9-12 days, and spray pure water every 3-5 days. S4-3. After the sporophyte seedlings are formed, spray nutrient solution 2, which consists of 1 / 2-1 modified Hoglund and 8-10g / L of rock sage extract, every 8-10 days, and spray pure water every 5-7 days. S5. When the sporophyte seedlings have grown 2-3 leaves or the plant height is 2-4cm, transplant them into seedling trays for cultivation.

[0006] Preferably, the method for collecting pure spores of *Ceratophyllum demersum* in step S1 is to collect mature pinnate leaves with brown sporangia on the *Ceratophyllum demersum* plant, place them in a clean sulfuric acid paper bag and air dry them naturally. After the spores naturally disperse, they are passed through a 200-mesh sieve to obtain pure spores, which are then stored in a refrigerator at 4°C for up to one year.

[0007] Preferably, the modified Hoglund nutrient solution has the following formula: potassium chloride 152.828 mg / L, dipotassium hydrogen phosphate trihydrate 456.638 mg / L, magnesium sulfate heptahydrate 246.48 mg / L, sodium iron EDTA 11.17 mg / L, boric acid 1.546 mg / L, manganese sulfate monohydrate 0.338 mg / L, copper sulfate 0.125 mg / L, zinc sulfate monohydrate 0.576 mg / L, and sodium molybdate 0.102 mg / L.

[0008] Preferably, the preparation method of the *Salvia splendens* extract in step S2 includes the following steps: S2-1. After crushing the rock-grown sage, soak it in an organic solvent for 10-25 minutes, then heat it at 60-80℃ and reflux for 30-60 minutes to obtain a crude extract. S2-2. After filtering the crude extract, retain the filtrate, concentrate it, and dry it to obtain the rock-grown sage extract.

[0009] Preferably, the organic solvent used in step S2-1 is an ethanol or ethyl acetate solution with a mass fraction of 50%-70%.

[0010] Preferably, the thickness of the spreading material evenly applied to the seedling container in step S3 is 2-3 cm.

[0011] Preferably, the sowing method in step S4 involves placing the spores of *Cibotium barometz* on sulfuric acid paper, gently tapping the paper to evenly scatter the spores onto the surface of the caterpillar-sphagnum moss substrate, and controlling the sowing amount to be 90-120 mg / m³. 2 .

[0012] Preferably, the substrate set in the seedling tray in step S5 is prepared by mixing peat moss, perlite and laterite in a volume ratio of 3:1:1.

[0013] This invention provides a method for efficiently cultivating Cibotium barometz using caterpillar-infested sphagnum moss, which has the following advantages compared to existing technologies: This invention utilizes caterpillar sphagnum moss in combination with modified Hoglund's and rock-dwelling sage extracts as a nutrient solution and subsequent foliar spray, effectively improving the germination rate and seedling survival rate of *Cibotium barometz* spores, while reducing contamination rates and shortening the seedling time to a certain extent, requiring only 115-119 days from sowing to obtaining transplantable seedlings. Furthermore, the method of this application does not require complex tissue culture equipment or aseptic operation, and frequent transfer and watering are unnecessary after sowing, making it suitable for large-scale, industrialized seedling production. The caterpillar sphagnum moss substrate is soft, minimizing root damage during transplanting, and the moss can be transplanted along with the seedlings, achieving a survival rate of over 94%. Attached Figure Description

[0014] Figure 1 This is the complete growth process of *Ceratophyllum demersum* spores germinating into sporophyte seedlings in Example 2 of the present invention; wherein A is *Ceratophyllum demersum* spores; B is *Ceratophyllum demersum* developing into a filamentous state; C is *Ceratophyllum demersum* developing into a platy state; D is *Ceratophyllum demersum* developing into a prothallus; E is *Ceratophyllum demersum* developing into a sporophyte; F is *Ceratophyllum demersum* plant after transplanting; Figure 2 The images show the spore germination of *Cyprinus fragilis* in Examples 2-4 and Comparative Example 1 of this invention; where A represents the overall macroscopic appearance of *Cyprinus fragilis* spore germination in different groups of culture boxes; and B represents a microscopic magnified observation of the young prothallus of *Cyprinus fragilis* spore germination in different groups. Figure 3 The images show the growth status of prothallus in different groups of *Cibotium barometz* according to the present invention; where A is a schematic diagram of the early stage of prothallus development using the *Sphagnum moss* substrate in Example 2; B is a schematic diagram of the mature stage of prothallus development using the *Sphagnum moss* substrate in Example 2; C is a schematic diagram of the early stage of prothallus development using the peat substrate in Comparative Example 2; D is a schematic diagram of the mature stage of prothallus development using the peat substrate in Comparative Example 2; E is a schematic diagram of the early stage of prothallus development using the peat + red clay substrate in Comparative Example 3; and F is a schematic diagram of the mature stage of prothallus development using the peat + red clay substrate in Comparative Example 3. Figure 4 This is a schematic diagram showing the growth of the transplanted golden dog fern in Embodiment 2 of the present invention. Detailed Implementation

[0015] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions of the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0016] Caterpillar sphagnum moss, the standard variety being Chilean red sphagnum moss, is named for its long, thick, segmented, and curled fibers resembling caterpillars, distinguishing it from the fine, fragmented common sphagnum moss found in the market. Its fibers are intact and fluffy, with a stable pore structure, exhibiting excellent water retention and air permeability, making it a superior performer. Under humid cultivation conditions, it is not prone to rotting or gelatinization, nor does it easily breed bacteria and algae, making it ideal for the growth of *Cibotium barometz* spores. Cultivation must be carried out entirely in a sterile environment. This raw material is readily available in the market and can be propagated on a large scale through horticultural methods. The caterpillar sphagnum moss used in the following examples was purchased from the Taobao store "Yongchun Horticulture".

[0017] Example 1: 1. Improve the preparation of Hoglund nutrient solution: The modified Hoglund nutrient solution was prepared according to the following components: potassium chloride 152.828 mg / L, dipotassium hydrogen phosphate trihydrate 456.638 mg / L, magnesium sulfate heptahydrate 246.48 mg / L, sodium iron EDTA 11.17 mg / L, boric acid 1.546 mg / L, manganese sulfate monohydrate 0.338 mg / L, copper sulfate 0.125 mg / L, zinc sulfate monohydrate 0.576 mg / L, sodium molybdate 0.102 mg / L (the remainder being distilled water); The prepared modified Hoglund nutrient solution was dispensed and autoclaved at 116°C for 30 minutes for later use.

[0018] 2. Preparation of extract from *Salvia splendens*: The crushed Salvia miltiorrhiza was soaked in a 60% ethanol solution for 20 minutes, and then extracted by reflux at 70°C for 45 minutes to obtain a crude extract. After filtering the crude extract to remove solid residue, a rock-grown sage extract was obtained. The extract of *Salvia solani* was obtained by concentrating and drying the extract.

[0019] Example 2: A method for efficiently cultivating Cibotium barometz using caterpillar-infested sphagnum moss: (1) Spore collection and pretreatment: In October 2025, spores of the golden dog fern were collected. Mature pinnae with brown sporangia and partially split sporangia were selected from the plant. After being cut off, they were placed in a clean sulfuric acid paper bag and placed in a well-ventilated and dry room to air dry naturally for 7 days. After the spores were naturally dispersed, the mixture in the paper bag was passed through a 200-mesh sieve to remove branches and leaves and impurities, and pure spore powder was obtained. It was then placed in a centrifuge tube and stored in a refrigerator at 4°C for later use.

[0020] (2) Preparation of seedling substrate: ① Sterilization: Immerse the caterpillar sphagnum moss in a sodium hypochlorite solution with an effective chlorine concentration of 1.0% for 15 minutes, stirring 3 times during the process to ensure full penetration of the solution; after pouring out the disinfectant, rinse repeatedly with plenty of purified water 5 times until the chlorine odor is no longer noticeable, then soak in an aqueous solution containing 0.5 g / L vitamin C (ascorbic acid) for 5 minutes to neutralize the residual chlorine, and finally rinse 3 times with purified water and drain for later use; ② Soaking in nutrient solution: Soak the sterilized sphagnum moss in a nutrient solution consisting of 1 / 2 times the amount of modified Hoglund nutrient solution and 10 g / L of rock sage extract for 3 hours until it has fully absorbed water. Remove it, squeeze it dry until it no longer drips, and spread it evenly in a seedling box with a length of 19 cm × 14 cm and a thickness of about 3 cm. Lightly compact it for later use. (3) Spore sowing: Remove the spore powder from the 4℃ refrigerator and allow it to return to room temperature. Weigh approximately 2.67 mg of spore powder (approximately 100 mg / m² based on a seedling tray area of ​​approximately 0.0266 m²) and place it on a clean sheet of parchment paper. Hold the paper and gently tap its surface to evenly distribute the spore powder onto the surface of the caterpillar sphagnum moss substrate. After sowing, lightly spray sterile water with a spray bottle to ensure the spores adhere closely to the surface of the caterpillar sphagnum moss. Cover the seedling tray with the lid to maintain humidity, leaving ventilation holes in the lid. (4) Seedling environment management: Place the seedling trays in a controllable greenhouse, and regulate the temperature to 25℃, relative humidity to 75%, and shading rate to 75% for daily management: From sowing until the gametophytes are visible to the naked eye, spray pure water every 4 days. Each time, spray the amount of water until the surface of the caterpillar sphagnum moss is moist but not waterlogged, and keep the substrate moist. From the time the gametophyte becomes visible to the naked eye until the sporophyte seedlings are formed, spray nutrient solution 1, consisting of 1 / 4 modified Hoglund + 5g / L rock sage extract, every 10 days, and spray pure water every 5 days until the surface of the caterpillar sphagnum moss is moist. After the sporophyte seedlings are formed, spray with nutrient solution 2 consisting of 1 / 2 times modified Hoglund + 10g / L rock sage extract every 8 days, and spray with pure water every 6 days.

[0021] (5) Seedling transplanting: When the sporophyte seedlings have grown 3 leaves or the plant height reaches 4cm, gently remove the seedlings from the caterpillar sphagnum moss and transplant them into 32-cell seedling trays. The substrate of the seedling trays is a mixture of peat moss, perlite, and red soil (volume ratio 3:1:1). After transplanting, water thoroughly to settle the roots and place the seedlings under the same greenhouse conditions, providing shade for 7 days, after which normal management can begin.

[0022] Example 3: A method for efficiently cultivating Cibotium barometz using caterpillar-infested sphagnum moss: The cultivation method in this embodiment is basically the same as that in Example 2, except for step (2): Step (2) of this embodiment is as follows: ① Sterilization: Place the caterpillar sphagnum moss into a sodium hypochlorite solution with an effective chlorine concentration of 1.0%, and soak it completely for 15 minutes, stirring 3 times during the process to ensure that the solution fully penetrates; after pouring out the disinfectant, rinse it repeatedly with a large amount of purified water 5 times until the chlorine smell is no longer noticeable, then soak it in an aqueous solution containing 0.5 g / L vitamin C (ascorbic acid) for 5 minutes to neutralize the residual chlorine, and finally rinse it 3 times with purified water and drain it for later use; ② Soaking in nutrient solution: Soak the sterilized caterpillar sphagnum moss in a nutrient solution consisting of 1 / 5 modified Hoglund and 7g / L rock sage extract for 3 hours. Squeeze it dry until it no longer drips, spread it evenly in a 19cm×14cm seedling box with a thickness of about 3cm, and lightly compact it for later use.

[0023] Example 4: A method for efficiently cultivating Cibotium barometz using caterpillar-infested sphagnum moss: The cultivation method in this embodiment is basically the same as that in Example 2, except for step (4): Step (4) of this embodiment is as follows: Place the seedling tray in a controllable greenhouse, and adjust the temperature to 25°C, relative humidity to 75%, and shading rate to 75%. Daily management is as follows: From sowing until the gametophytes are visible to the naked eye, spray pure water every 4 days. Each time, spray enough to moisten the surface of the caterpillar sphagnum moss but not to accumulate water, keeping the substrate moist.

[0024] From the time the gametophyte becomes visible to the naked eye until the sporophyte seedlings are formed, spray nutrient solution 1, consisting of 1 / 4 modified Hoglund + 7g / L rock sage extract, every 10 days, and spray pure water every 5 days until the surface of the caterpillar sphagnum moss is moist. After the sporophyte seedlings are formed, spray with nutrient solution 2 consisting of 1 / 2 times modified Hoglund + 8g / L rock sage extract every 8 days, and spray with pure water every 6 days.

[0025] Comparative Example 1: A method for efficiently cultivating Cibotium barometz using caterpillar-infested sphagnum moss: The cultivation method of this comparative example is basically the same as that of Example 2, except that steps (2) and (4) are different: The steps (2) of this comparative example are as follows: ① Sterilization: Place the caterpillar sphagnum moss into a sodium hypochlorite solution with an effective chlorine concentration of 1.0%, and soak it completely for 15 minutes, stirring 3 times during the process to ensure that the solution fully penetrates; after pouring out the disinfectant, rinse it repeatedly with a large amount of purified water 5 times until the chlorine smell is no longer detectable, then soak it in an aqueous solution containing 0.5 g / L vitamin C (ascorbic acid) for 5 minutes to neutralize the residual chlorine, and finally rinse it 3 times with purified water and drain it for later use; ②Soaking: Soak the sterilized caterpillar sphagnum moss in pure water for 3 hours until it is fully saturated. Remove it, squeeze it dry until it stops dripping, and spread it evenly in a 19 cm × 14 cm seedling box with a thickness of about 3 cm. Lightly compact it for later use. Step (4) is as follows: Place the seedling tray in a controllable greenhouse and adjust the temperature to 25℃, relative humidity to 75%, and shading rate to 75%. From sowing to the formation of sporophyte seedlings, spray pure water once every 4 days. The amount of water sprayed each time should be enough to moisten the surface of the caterpillar sphagnum moss but not to accumulate water, so as to keep the substrate moist.

[0026] Comparative Example 2: A method for efficiently cultivating Cibotium barometz using caterpillar-infested sphagnum moss: The cultivation method of this comparative example is basically the same as that of Example 2, except that steps (2) and (4) are different: The steps (2) of this comparative example are as follows: ① Sterilization: Place the caterpillar sphagnum moss into a sodium hypochlorite solution with an effective chlorine concentration of 1.0%, and soak it completely for 15 minutes, stirring 3 times during the process to ensure that the solution fully penetrates; after pouring out the disinfectant, rinse it repeatedly with a large amount of purified water 5 times until the chlorine smell is no longer detectable, then soak it in an aqueous solution containing 0.5 g / L vitamin C (ascorbic acid) for 5 minutes to neutralize the residual chlorine, and finally rinse it 3 times with purified water and drain it for later use; ② Soaking in nutrient solution: Soak the sterilized sphagnum moss in 1 / 2 times the amount of modified Hoglund nutrient solution for 3 hours until it has fully absorbed water. Remove it, squeeze it dry until it no longer drips, and spread it evenly in a seedling box with a length of 19 cm × 14 cm and a thickness of about 3 cm. Lightly compact it for later use. Step (4) is as follows: Place the seedling trays in a controlled greenhouse, adjust the temperature to 25℃, the relative humidity to 75%, and the shading rate to 75%, and carry out daily management: From sowing until the gametophytes are visible to the naked eye, spray pure water once every 4 days. Each time, spray the amount of water until the surface of the caterpillar sphagnum moss is moist but not waterlogged, and keep the substrate moist. From the time the gametophyte becomes visible to the naked eye until the sporophyte seedlings are formed, spray with 1 / 4 of the modified Hoglund nutrient solution every 10 days and with pure water every 5 days, until the surface of the caterpillar sphagnum moss is moist. After the sporophyte seedlings are formed, spray with 1 / 2 times the modified Hoglund nutrient solution every 8 days and with pure water every 6 days.

[0027] Comparative Example 3: Methods for cultivating golden dog ferns in peat moss: The cultivation method of this comparative example is basically the same as that of Example 2, except for steps (2) and (4).

[0028] Step (2): Use peat moss as the seedling substrate. Moisten the peat moss with a nutrient solution consisting of 1 / 2 modified Hoglund and 10g / L rock sage extract until it can be formed into a ball by hand and crumbled easily when rubbed. Fill the seedling box with peat moss to a thickness of about 3cm, compact and flatten it with a flat plate, and set aside for later use.

[0029] Step (4): Place the seedling trays in a controlled greenhouse, setting the temperature to 25℃, relative humidity to 75%, and shading rate to 75%, and carry out daily management. Since peat moss has lower water retention than caterpillar sphagnum moss, the frequency of watering and nutrient solution application needs to be increased accordingly to maintain substrate moisture.

[0030] From sowing until the gametophytes are visible to the naked eye, spray with pure water once a day, the amount of water sprayed each time should be enough to moisten the surface of the substrate but not to cause water accumulation.

[0031] From the time the gametophyte becomes visible to the naked eye until the sporophyte seedlings are formed, spray a nutrient solution consisting of 1 / 4 modified Hoglund + 5g / L rock sage extract every 10 days, and spray pure water every 2 days until the surface of the caterpillar sphagnum moss is moist. After the sporophyte seedlings are formed, spray with a nutrient solution consisting of 1 / 2 times modified Hoglund + 10g / L rock sage extract every 8 days, and spray with pure water every 2-3 days.

[0032] Comparative Example 4: Method for cultivating golden dog fern by mixing peat moss and red soil: The cultivation method of this comparative example is basically the same as that of comparative example 2, except for step (2).

[0033] Step (2): Cultivate golden dog spores using a mixed substrate prepared by mixing peat moss and red soil at a volume ratio of 5:1. Moisten the mixed substrate with a nutrient solution consisting of 1 / 2 modified Hoglund and 10 g / L rock sage extract until it can be formed into a ball by hand but crumbles easily when rubbed. Fill the substrate into a 54 cm × 28 cm seedling box, with a substrate thickness of about 3 cm, and compact and flatten it with a flat plate before use.

[0034] Detection: Golden-haired dog ferns were cultivated according to the methods of Examples 2-4 and Comparative Examples 1-4 above, with other conditions being the same and suitable for the growth of golden-haired dog ferns. The development of golden-haired dog ferns was observed, and the time of spore germination, prothallus formation and sporophyte appearance were recorded. The contamination of the substrate during the cultivation process was also observed. The results are shown in Table 1.

[0035] Table 1. Development of *Cibotium barometz* in different embodiments and comparative examples.

[0036] In Table 1 above: Spore germination (days) refers to the number of days required from the day of sowing to spore germination; prothallus maturation (days) refers to the number of days required from sowing to the appearance of mature prothallus; sporophyte (days) refers to the number of days required from sowing to the appearance of sporophyte; contamination status refers to the degree of contamination by ferns, moss, or mold. Among the contamination statuses, no finger contamination rate is 0%; slightly contaminated finger contamination rate is less than 5%; moderately contaminated finger contamination rate is 10-20%; moderately contaminated finger contamination rate is 30-50%; and severely contaminated finger contamination rate is greater than 50%. The contamination rate is calculated as follows: (Contaminated substrate area / Total substrate area). Because the spores of *Cibotium barometz* are small and numerous, it is impossible to statistically determine the germination rate; the prothallus and sporophyte are also numerous, making statistical analysis difficult. Therefore, different quantities of... The symbol indicates that the germination rate is calculated based on the coverage of the leaflets in the seedling tray. This indicates a coverage rate greater than 90%. This indicates a coverage rate of 80%-90%. This indicates a coverage rate of 60%-79%. This indicates a coverage rate of 30%-59%. This indicates that the coverage is less than 30%, and the germination rate is calculated 30 days after sowing.

[0037] Number of protophylls: The number of protophylls is calculated based on the coverage of protophylls in the seedling tray. This indicates a coverage rate greater than 90%. This indicates a coverage rate of 80%-90%. This indicates a coverage rate of 60%-79%. This indicates a coverage rate of 30%-59%. This indicates a coverage of less than 30%; the prothallus count is performed 15 days after the prothallus begins to appear.

[0038] Sporophyte count: The sporophyte count is calculated based on the number of young spore seedlings per unit area. This indicates that there are more than 200 seedlings per 100 square centimeters of substrate. This indicates that there are 150-200 seedlings per 100 square centimeters of substrate. This indicates that there are 100-149 seedlings per 100 square centimeters of substrate. This indicates that there are 50-99 seedlings per 100 square centimeters of substrate. This indicates that the number of young spore seedlings per 100 square centimeters of substrate is less than 50. The sporophyte count is performed 30 days after the first appearance of sporophytes.

[0039] Table 2. Total seedling time and transplant survival rate of *Cibotium barometz* in different embodiments and comparative examples.

[0040] The time required from sowing to sporophyte in Table 2 above refers to the total time required from sowing spores to the appearance of 2-3 leaves or when the plant reaches a height of 2-4 cm.

[0041] As can be seen from Tables 1 and 2, the method of using the caterpillar sphagnum moss combined with nutrient solution as a sowing substrate for the efficient cultivation of golden ferns can significantly shorten the cultivation cycle. The incidence of contamination by miscellaneous bacteria and ferns is zero throughout the process, and the number of sporophytes is greatly increased. Its overall effect is significantly better than that of the other pairs.

[0042] Comparing Examples 2-4 with Comparative Examples 1-2, it can be seen that when using caterpillar sphagnum moss substrate to cultivate Cibotium barometz, the treatment group without added nutrient solution only formed a small number of sporophytes, indicating that the nutrient solution composed of modified Hoglund's and rock sage extracts can significantly improve the sporophyte development of Cibotium barometz.

[0043] The comparison results between Examples 2-4 and Comparative Examples 3-4 show that using caterpillar sphagnum moss combined with nutrient solution as a seedling substrate significantly outperformed peat moss and a mixture of peat moss and red soil (5:1) in the spore cultivation process of *Ceratophyllum demersum*. Specifically, in Examples 2-4, the spore germination time, prothallium maturation time, and sporophyte emergence time were all earlier than in Comparative Examples 3-4, and the time required from spore sowing to sporophyte transplantation was 115-119 days, significantly shortening the seedling cycle. Furthermore, in Comparative Examples 3-4, approximately 45 days after sowing (i.e., the initial stage of prothallium maturation) often saw the proliferation of large amounts of moss and other ferns, while this phenomenon was not observed in Examples 2-4. Because moss and ferns grow quickly, they rapidly take over the growth space and nutrients of the golden dog fern. As a result, although the spore germination rate is high, most prothallia are covered by moss in the early stages of development, and only a few can develop into mature prothallia, which in turn significantly reduces the number of sporophytes formed in the end.

[0044] This embodiment does not require hardening off the seedlings, and the transplant survival rate can reach 94%.

[0045] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for efficiently cultivating *Cibotium barometz* using caterpillar-infested sphagnum moss, characterized in that, The method includes the following steps: S1. Collect pure spores of the golden-haired dog fern for later use; S2. After diluting the modified Hoglund nutrient solution to 1 / 10-1 / 2, add rock sage extract to adjust the content to 7-14 g / L to obtain the nutrient solution. The rock sage extract is obtained by extracting rock sage with organic solvent by heating and reflux, filtering, and concentrating and drying the filtrate. The modified Hoglund nutrient solution has the following formula: potassium chloride 152.828 mg / L, dipotassium hydrogen phosphate trihydrate 456.638 mg / L, magnesium sulfate heptahydrate 246.48 mg / L, sodium iron EDTA 11.17 mg / L, boric acid 1.546 mg / L, manganese sulfate monohydrate 0.338 mg / L, copper sulfate 0.125 mg / L, zinc sulfate monohydrate 0.576 mg / L, and sodium molybdate 0.102 mg / L. S3. After sterilizing the caterpillar sphagnum moss, soak it in the above nutrient solution for 2-4 hours, then take it out and squeeze it dry until it no longer drips, and then spread it evenly in the seedling container to obtain the caterpillar sphagnum moss substrate for later use. S4. Evenly spread the pure spores of *Cibotium barometz* onto the surface of the *Sphagnum moss* substrate. Spray with sterile water to ensure close adhesion between the spores and the *Sphagnum moss* surface. Then cover the substrate and place it in an environment with a temperature of 25±2℃, relative humidity of 70%-80%, and shading rate of 70%-80% for routine management until the spores germinate and form sporophyte seedlings. The specific methods for routine management include the following steps: S4-1. From sowing until the gametophytes are visible to the naked eye, spray pure water every 3-5 days to keep the caterpillar sphagnum moss substrate moist but not waterlogged. S4-2. From the time the gametophyte becomes visible to the naked eye until the sporophyte seedlings are formed, spray nutrient solution 1, which consists of 1 / 4-1 / 2 modified Hoglund and 5-7 g / L of rock sage extract, every 9-12 days, and spray pure water every 3-5 days. S4-3. After the sporophyte seedlings are formed, spray with nutrient solution 2, which consists of 1 / 2-1 modified Hoglund and 8-10g / L of rock sage extract, every 8-10 days, and spray with pure water every 5-7 days. S5. When the sporophyte seedlings have grown 2-3 leaves or the plant height is 2-4cm, transplant them into seedling trays for cultivation.

2. The method according to claim 1, characterized in that: The method for collecting pure spores of *Cereus undatus* in step S1 is to collect mature pinnate leaves with brown sporangia on the *Cereus undatus* plant, put them in a clean sulfuric acid paper bag and let them air dry naturally. After the spores are naturally dispersed, they are passed through a 200-mesh sieve to obtain pure spores, which are then stored in a refrigerator at 4°C for up to one year.

3. The method according to claim 1, characterized in that: The modified Hoglund nutrient solution has the following formula: potassium chloride 152.828 mg / L, dipotassium hydrogen phosphate trihydrate 456.638 mg / L, magnesium sulfate heptahydrate 246.48 mg / L, sodium iron EDTA 11.17 mg / L, boric acid 1.546 mg / L, manganese sulfate monohydrate 0.338 mg / L, copper sulfate 0.125 mg / L, zinc sulfate monohydrate 0.576 mg / L, and sodium molybdate 0.102 mg / L.

4. The method according to claim 1, characterized in that, The preparation method of the rock-grown sage extract in step S2 includes the following steps: S2-1. After crushing the rock-grown sage, soak it in an organic solvent for 10-25 minutes, then heat it at 60-80℃ and reflux for 30-60 minutes to obtain a crude extract. S2-2. After filtering the crude extract, retain the filtrate, concentrate it, and dry it to obtain the rock-grown sage extract.

5. The method according to claim 4, characterized in that: The organic solvent used in step S2-1 is an ethanol or ethyl acetate solution with a mass fraction of 50%-70%.

6. The method according to claim 1, characterized in that: In step S3, the thickness of the spreading material evenly applied to the seedling container is 2-3 cm.

7. The method according to claim 1, characterized in that: In step S4, the sowing method involves placing the spores of *Ceratophyllum demersum* on sulfuric acid paper, gently tapping the paper to evenly scatter the spores onto the surface of the caterpillar-infested sphagnum moss substrate, and controlling the sowing amount to be 90-120 mg / m³. 2 .

8. The method according to claim 1, characterized in that, The substrate set in the seedling tray in step S5 is prepared by mixing peat moss, perlite and red clay in a volume ratio of 3:1:1.

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