CN119605553A

CN119605553A - A method for breeding golden dog spores and rejuvenating young spore seedlings

Info

Publication number: CN119605553A
Application number: CN202510008408.1A
Authority: CN
Inventors: 李娟�; 周绍容; 汤猛; 陈利君; 严岳鸿
Original assignee: ORCHID CONSERVATION & RESEARCH CENTER OF SHENZHEN
Current assignee: ORCHID CONSERVATION & RESEARCH CENTER OF SHENZHEN
Priority date: 2024-12-23
Filing date: 2025-01-03
Publication date: 2025-03-14

Abstract

The present invention provides a method for breeding Cibotium barometz spores and rejuvenating young spore seedlings. The method for artificially cultivating Cibotium barometz provided by the present invention comprises the following steps: transplanting the Cibotium barometz spore seedlings to be rejuvenated into a matrix for rejuvenation culture; wherein the matrix is a mixture of leaf mold and peat soil in a volume ratio of (1-1.5):1 or 1:1, or a mixture of leaf mold, white vermiculite, and garden soil in a volume ratio of 4:1:3. The method has low cost, simple technology, high yield, fast seedling growth rate, and is suitable for large-scale batch production. It can effectively meet the rapid breeding needs of Cibotium barometz and can be widely used in the fields of landscaping, medicinal production, and scientific research.

Description

Method for breeding and rejuvenating golden hair dog spores and young spore seedlings

Technical Field

The invention relates to the technical field of forest seedling cultivation, in particular to a method for breeding and rejuvenating a trichocarpa lobayensis spore.

Background

The golden hair dog (Cibotium barometz (Linn.) J.Sm.) belongs to the genus golden hair dog (Cibotium) of the family golden hair dog (Cibotiaceae), is a perennial high tree fern medicinal plant, and is mainly distributed in Asian tropical and subtropical areas, especially in south China. The golden hair dog has thick and thick root stem, is in a lying state, has a cluster of big leaves at the top, brown petiole and covered with pad-shaped golden hair at the base. The light scattering type ornamental plant is favorable for scattering light, is often used in a wet and warm woody environment, and is an internationally known ornamental plant for gardens and important traditional Chinese medicinal materials. Gold hair dogs are known for their unique appearance and medicinal value. The plant type is tall and tall, the leaves are large, the appearance is attractive, the ornamental value is extremely high, and the plant type plays an important role in gardening application. The rhizome (Chinese medicinal material named as 'rhizoma Cibotii') is recorded in Chinese pharmacopoeia (2020 edition), has effects of dispelling pathogenic wind and dampness, nourishing liver and kidney, strengthening waist and knee, etc., and is commonly used for treating rheumatalgia, soreness of waist and knee, weakness of lower limbs, etc. The scaly hair on the rhizome can also be used for external application to stop bleeding. In addition, in recent years, a plurality of active ingredients (such as flavonoids and polysaccharide compounds) are extracted from the golden hair dogs through a modern process, a new development direction is provided for the pharmaceutical technology, and the health care edible value of the golden hair dogs is also gradually valued.

Although the golden hair dog has great ornamental, medicinal and health care values, the wild resources of the golden hair dog are reduced year by year due to unordered development. In recent years, due to the excessive digging of wild resources of the wild dogs, the growth rule and the distribution characteristic of the wild dogs are ignored, so that the number of the wild dogs is drastically reduced. Currently, the golden hair dogs have been listed in the international trade convention of endangered wild animal and plant species (appendix II, CITES 2016) and simultaneously in the national secondary protection plants of the national importance protection wild plant directory (first lot). Its endangered condition is mainly due to the difficulty of reproduction, slow growth, and the extensive and lacking proprietary management techniques for ferns in traditional planting methods. The current planting method is not ideal in the aspects of seedling survival rate, seedling growth speed, plant quality and the like, so that the development of the golden hair dogs depends on wild resources, and the problem of resource shortage is further aggravated. With the increasing demand for the resources of the golden hair dogs in the fields of gardening, medicine and scientific research, the protection of wild golden hair dog resources and the development of effective propagation methods have become urgent.

The propagation of the rhizoma cibotii mainly comprises three modes of sexual propagation (spore propagation), asexual propagation (plant division propagation) and in vitro propagation (tissue culture). The problems of low germination rate, uneven emergence of seedlings, slow growth of seedlings, high proportion of weak seedlings and the like often cause long seedling raising period, low survival rate, poor adaptability of seedlings and even large-area dead seedlings, and severely restrict the popularization and application of artificial large-scale breeding of the golden hair dogs, so that the method becomes a key technical bottleneck to be solved urgently at present.

Disclosure of Invention

In order to solve the problem of shortage of the gold hair dog resources, the inventor of the invention obtains a set of artificial breeding method for breeding the gold hair dog through a great deal of fuzziness, and the method has high survival rate and short time consumption. The method of the invention provides a solution with the characteristics of regular and quick emergence and strong growth aiming at the difficult seedling raising problems of low germination rate of the golden hair dog spores, uneven emergence of gametophytes, yellow sporophytes and the like.

The method for artificially culturing the golden hair dog (Cibotiumbarometz) comprises the following steps of planting golden hair dog sporophyte seedlings to be rejuvenated into a matrix for rejuvenation culture, wherein the matrix is a mixture of leaf rot soil and peat soil in a volume ratio of (1-1.5) to (1) or (1:1), or a mixture of leaf rot soil, white vermiculite and garden soil in a volume ratio of (4:1:3).

The leaf-rotting soil, also called humus soil, is nutrient soil formed by decomposing and fermenting plant branches and leaves in soil through microorganisms. As a specific example, the soil with the rotten leaves has the organic matter content of 45.7%, the soil volume weight of 0.36g/cm ³, the PH (water immersion) of 6.0 and the soil with the rotten leaves has black and organic matter particles.

The peat soil (peat soil) is soil which is formed by accumulating a large amount of insufficiently decomposed plant residues and forming a peat layer under the condition of hypoxia because of long-term ponding and dense aquatic vegetation in certain river and lake sedimentary low plain and valley between mountains. As a specific example, the sum of organic matters and humic acid in peat soil is 50% or more, the porosity is 70-80, and the PH (water immersion) is 5.6-6.5.

Preferably, in the above method, when the first true leaf of the sporophyte seedling grows to 0.3-0.5cm, the transplanting and rejuvenation culture are performed.

Preferably, the rejuvenation culture conditions comprise a temperature of 23-25 ℃, an illumination intensity of 2500-2500 Lux, an illumination time of 12-14 hours or 14 hours per day, and an air relative humidity of 65-85% of a space where sporophyte seedlings are located. Wherein the space where the sporophyte seedlings are located is a space formed by transplanting the sporophyte seedlings in a seedling raising container and covering the seedling raising container with a transparent cover.

Preferably, the water content of the matrix in the rejuvenation culture is 60% -65% or 60%.

Preferably, the rejuvenation culture further comprises a step of applying a nutrient solution, wherein the nutrient solution is a macroelement water-soluble fertilizer, and further specifically, the nutrient solution is applied from the 90 th day after the transplanting, the macroelement water-soluble fertilizer is diluted into an aqueous solution with the concentration of 0.2% (mass percent concentration), and 200ml of the aqueous solution is sprayed every 7 days and every 1760 square centimeters.

Among them, macroelement water-soluble fertilizers are the type of fertilizers commonly used in plant growth, which are mainly composed of macroelements nitrogen (N), phosphorus (P) and potassium (K) necessary for three plants. These elements play a key role in the growth and development of plants and are widely used in agricultural production such as drip irrigation, spray application, foliar fertilization, etc. because they are completely dissolved in water for plant absorption, the proportions used are balanced, which contain 20% nitrogen (N), 20% phosphorus (P ₂O₅) and 20% potassium (K ₂ O) (model: 20-20-20+TE, implementation standard NY/T1107-2020).

Preferably, the method further comprises the step of hardening off, wherein sporophyte seedlings to be hardened off are separately planted in a separated mode, and the matrix is a mixture of leaf rot soil and peat soil in a volume ratio of (1-1.5): 1 or 1:1.

Preferably, the sporophyte seedlings to be acclimatized are sporophyte seedlings with a height of 8.5-10.0cm or sporophyte seedlings with 5-8 leaves per plant.

Preferably, the seedling hardening conditions comprise the temperature of 25-30 ℃, the air humidity in a greenhouse of 60-65%, the illumination intensity of the seedling hardening of 3000-3700 lux and the illumination time of 12-14 h.

Preferably, the method comprises the steps of seeding spores of the golden hair dog (Cibotiumbarometz) in a substrate, performing a dark treatment, and culturing until sporophyte seedlings to be rejuvenated are obtained.

Preferably, the dark treatment condition comprises culturing for 44-48 h or 48h under the dark condition, specifically, the dark treatment condition comprises culturing for 44-48 h or 48h under the dark and temperature of 23-25 ℃, further specifically, the dark treatment condition comprises culturing for 44-48 h or 48h under the dark and substrate initial humidity of 60% or 60-65%, the temperature of 23-25 ℃ and the airtight condition, and still further specifically, the dark treatment condition comprises culturing for 44-48 h or 48h under the dark and substrate initial humidity of 60% or 60-65%, the temperature of 23-25 ℃ and the airtight condition without water supplementing. The sealing means that a transparent cover is covered on the seedling raising container, and then a gap between the container and the cover is sealed to form a sealed environment space, wherein spores or sporophyte seedlings are all positioned in the seedling raising container.

Preferably, the step of re-culturing until the sporophyte seedlings to be rejuvenated are obtained comprises the step of culturing until the gametophyte sexual organs are mature under the conditions that the ambient temperature is 23-25 ℃, the illumination intensity is 2500-3500 Lux, the illumination is 12-14 hours or 14 hours per day, and the relative air humidity of the environment where the gametophyte is located is 65-85%. Specifically, the culture is carried out under the conditions that the temperature of the environment is 23-25 ℃, the illumination intensity is 2500-2500 Lux, the illumination is carried out for 12-14 hours or 14 hours each day, the relative humidity of the air of the environment where the gametophyte is located is 65-85%, and the culture is sealed. Wherein, the relative humidity of the air in the environment where the gametophyte is positioned is the relative humidity of the air in the seedling raising container covered with the transparent cover. Wherein, the sealing means that the seedling raising container is covered with a transparent cover, and a gap between the container and the cover is sealed to form a space environment. If the relative humidity of the air is not within the range of 65-85%, water is supplemented to enable the relative humidity of the air to be 65-85%, and the air is immediately restored to a closed state after water is supplemented.

Preferably, after maturation of the gametophytic organ, the steps are included:

(1) The method comprises the steps of adjusting the relative humidity of air in an environment where a gametophyte is located to 65-85%, culturing for 10 days under the conditions of sealing and 25 ℃ constant temperature, wherein water is not added in the environment, specifically adjusting the relative humidity of air in the environment where the gametophyte is located to 65-85%, culturing for 10 days under the conditions of sealing and 25 ℃ constant temperature, illumination intensity of 2500-2500 Lux and illumination for 12-14 hours or 14 hours each day, and culturing for 10 days without water adding in the environment where the gametophyte is located, wherein the relative humidity of air in a seedling culture container covered with a transparent cover is the relative humidity of air. Wherein, the sealing means that the seedling raising container is covered with a transparent cover, and a gap between the container and the cover is sealed to form a space environment.

(2) And then the closed condition is relieved, the gametophyte is placed in a condition that the illumination intensity is 2500-2500 Lux, the illumination is carried out for 12-14 hours or 14 hours every day, the air relative humidity of the environment where the gametophyte is located is 22-25%, and the gametophyte is cultivated for 22-24 hours or 24 hours, wherein the air relative humidity of the environment where the gametophyte is located is 22-25%, namely the condition in a greenhouse, after the closed environment is relieved, the gametophyte is exposed to the greenhouse, and the air relative humidity of the greenhouse is 22-25%. Wherein the sealing condition is released, i.e. the transparent cover is taken off.

(3) Fertilization was promoted by water spray treatment.

Preferably, the method comprises the steps of culturing until sporophyte seedlings germinate after fertilization, further specifically culturing until sporophyte seedlings germinate under the conditions that the ambient temperature is 23-25 ℃, the illumination intensity is 2500-3500 Lux, the illumination is 12-14 hours or 14 hours each day, the relative air humidity is 65-85%, and still further specifically culturing is carried out under a closed condition. Wherein, the sealing means that the seedling raising container is covered with a transparent cover, and a gap between the container and the cover is sealed to form a space environment.

Preferably, after the sporophyte seedlings germinate, culturing under the condition that the illumination intensity is 3000-4000 Lux, the illumination is carried out for 12-14 hours or 14 hours each day, the temperature is 26-27 ℃, and the relative air humidity is kept at 55-70%, wherein the relative air humidity is the relative air humidity in a seedling culture container covered with a transparent cover.

Preferably, the substrate for spore inoculation is peat soil, in particular, peltier peat soil, the particle size of which is 0-6 mm, and the pH value of which is 5.5-6.0.

Preferably, the initial moisture content of the substrate for spore inoculation is 60%.

Here, sporophyte seedlings and young spore seedlings have the same meaning and are interchangeable with each other.

The method for raising the seedlings of the golden hair dogs can realize quick batch seedling raising of spores, obviously improves the germination rate of spores (to 3000 strains/disc, namely 3000 strains/0.176 square meter), has high seedling emergence uniformity and strong gametophyte sporophytes, and shortens the growth period of seedlings to 120 days. In a word, the method has the advantages of low cost, simple technology, high yield and high seedling growth speed, is suitable for large-scale mass production, can effectively meet the rapid breeding requirement of the golden hair dog, and can be widely applied to the fields of landscaping, medicinal production, scientific research and the like.

Drawings

FIG. 1 shows sporophylls of the dog.

FIG. 2 is a mature gametophyte of a gold hair dog.

FIG. 3 shows a seedling of a spore of a Cibotium barometz.

FIG. 4 shows the separately planted rejuvenation matrix for different treatments.

FIG. 5 shows the seedling growth on day 15 of T1 group.

FIG. 6 shows the growth and development of young spore seedlings on day 45 in different groups.

FIG. 7 shows the growth and development of the seedlings on day 90 of the T1 group.

FIG. 8 shows the growth and development of seedlings on day 120 of the T1 group.

FIG. 9 shows the growth and development of the day 150 seedlings of the golden hair dogs.

Detailed Description

1. Collecting the spores of the golden hair dog

The sporophore group cover cracking rate is selected to be 20% -30%, and the mature and well-grown aureobasidium sporophylls of the plant are collected. During collection, the sporophylls are placed into a rectangular paper bag made of 2-3 layers of newspapers, sealed, and then placed in a cool and ventilated place for natural drying for 10-15 days. Before sowing, the dried paper bag is lifted slightly, a corner is cut off, and spores are beaten and fall on white paper. If the spores are mixed with impurities, the spores can be uniformly spread, the white paper is inclined and flicked, so that the spores are separated from the impurities. After repeated treatments until impurities are completely removed, purified spores are collected into centrifuge tubes and stored in a4 ℃ refrigerator to ensure the activity and quality of spores.

2. Inoculation of golden hair dog spores

The soil used for sowing is Danish Inlet Pink peat soil (particle size 0-6 mm, pH value 5.5-6.0) provided by Hongda biological technology Co Ltd in Beijing. After the peat soil is wrapped by newspaper, sterilizing by high-pressure steam at 125 ℃ for 25 minutes, airing, spreading the peat soil in a culture dish of 55cm multiplied by 32cm multiplied by 8cm, compacting the peat soil to the soil layer thickness of 5cm, and spraying tap water until the soil water content is about 60%. When sowing, 0.01ml spores are placed at fold lines of folded A4 paper, and the paper is flicked, so that the spores are uniformly sown into the culture tray along the fold lines. After sowing, the seedling raising tray is covered by a transparent cover, gaps between the tray covers are sealed by a black electrical adhesive tape, the substrate is kept moist, and watering is not needed after sowing. Covering the seedling raising tray after seeding with opaque black cloth, and placing the seedling raising tray in a culture room with the environmental temperature of 23-25 ℃ for 48 hours so as to promote synchronous germination of spores, wherein the seedlings grow more uniformly and tidily. After the dark treatment, the temperature of the culture environment is controlled to be 23-25 ℃, the illumination intensity is 2500-2500 Lux, the illumination is carried out for 14 hours every day, the relative humidity of air (the relative humidity of air in the seedling raising tray) is kept to be 65-85% by timely supplementing water, namely, the matrix in the seedling raising tray is sprayed with water to enable the humidity in the seedling raising tray to reach 65-85%, the seedling raising tray is immediately covered by a transparent cover, gaps among the tray covers are sealed by an electric adhesive tape, and the sealed environment is kept until the following germination begins and then the cover is opened.

3. Artificial assisted fertilization and transformation of sporozoites

After 10-15 days after sowing, the golden hair dog spores start to germinate, and after about 60-80 days, gametophytic organs (spermatids and cervical ova) develop and mature, and the formation of the gametophytic organs can be observed through a microscope. After the sexual organ is mature, the cover is opened to detect the humidity, the water mist is sprayed to keep the humidity at 65-85%, then the temperature is kept at 25 ℃, the culture dish is sealed for 10 days, watering is not needed during the period, the illumination intensity is 2500-3500 Lux, and the illumination is carried out for 14 hours every day. Then, the transparent plastic cover is opened, the humidity is not kept by spraying, and the cover is naturally dried in a greenhouse for 24 hours (during the period, the temperature is 23-25 ℃, the illumination intensity is 2500-2500 Lux, the illumination is 14 hours per day, the relative humidity of air in a culture room is 22-25%), so that the sufficient development of a spermatid and the improvement of the sperm concentration are promoted. After drying, fertilization was completed by sufficiently impregnating the sex organs with water by a small number of uniform sprays (once each of the morning, noon, and evening). After fertilization, the environment is sealed again for 7 days, namely, the cultivation plate is covered by a transparent cover and sealed by an electric adhesive tape, water is not moved or sprayed in the period (the relative humidity of air in the cultivation plate is 65-85 percent in the period) until sporophyte seedlings germinate. If no seedling growth is seen after 7 days, the transparent cover can be opened for ventilation for 3 days, and then the spraying and sealing steps are repeated, wherein the temperature is 23-25 ℃ and the illumination intensity is 2500-2500 Lux in the whole fertilization process, and the illumination is carried out for 14 hours every day. After seedlings grow out, the illumination intensity is increased to 3000-4000 Lux, the illumination is carried out for 14 hours each day, the room temperature is controlled at 26-27 ℃, and the relative air humidity (the relative air humidity in a seedling raising tray) is kept at 55-70%. The germination rate of the golden hair dog spores can reach 3000 strains/disc (3000 strains/0.176 square meter) after the treatment.

4. Seedling rejuvenation

After the young dog sporophytes grow out, the growth is observed every 3 days. When the first true leaves grow to 0.3-0.5 cm, transplanting the seedlings into a culture tray with the same specification as that of sowing by forceps, and dividing and rejuvenating. The study shows that the separate planting matrix has obvious rejuvenation effect on young Cinnamomum kansui seedlings, in order to screen the optimal matrix, the test adopts leaf rot soil and peat soil as main materials, coconut bricks, white vermiculite (5-8 mm), pond sludge and garden soil as auxiliary materials, different matrix combinations are prepared (specific proportions are shown in table 1), and the water content of the matrix is 60%. 4 treatments (T1-T4) are set for the test, 3 trays of seedlings are transplanted per treated substrate, 100 plants are planted per tray, and 1 repetition is carried out per tray, and the total number of the seedlings is 3.

Coconut bricks, white vermiculite, pond sludge and garden soil are all purchased from Shenzhen Longpost district Huihuang gardening product shops.

The leaf rot soil, also called humus soil, is a nutrient soil formed by decomposing and fermenting plant branches and leaves in soil by microorganisms. In this example, leaf rot soil was purchased from Shenzhen green state city garden limited. The organic matter content is 45.7%, the soil volume weight is 0.36g/cm ³, the PH (water immersion) is 6.0, the rotten leaf soil texture is black, and the organic matter soil particles.

Peat soil (peat soil) refers to soil in which a large amount of insufficiently decomposed plant residues accumulate and form a peat layer under the condition of hypoxia because of long-term water accumulation and dense aquatic vegetation in certain river and lake sedimentary low plain and valley lands. In the embodiment, peat soil is purchased from vinca seedling peat science and technology Co., ltd, and the product name is novel seedling substrate-Jin Tianpai. The peat soil has a porosity of 70-80 and a PH (water immersion) of 5.6-6.5, wherein the sum of the organic matter and humic acid is 50% or more.

Table 1 composite seedling substrate formula (volume ratio)

The substrate used for the plant division rejuvenation does not need sterilization, and the plant division container is kept by a seedling raising tray and a transparent cover for sowing, but is not sealed by adhesive tape. After the separate planting, the culture temperature is controlled at 23-25 ℃, the illumination intensity is 2500-3500 Lux, and the illumination time is 14 hours per day. And supplementing water timely, keeping the relative humidity of air in the seedling raising tray at 65% -85%, and spraying water mist to improve the humidity when the humidity is lower than 65%, and properly ventilating when the humidity is higher than 85%. And observing the seedling reviving condition and the plant diseases and insect pests, and recording the survival rate (%) of the sporophyte seedlings on the 15 th day, the 30 th day, the 45 th day, the 60 th day, the 75 th day and the 90 th day after the sub-planting, wherein the survival rate is calculated as the number of survival plants/100 multiplied by 100%. The plant heights were measured on day 30, day 60 and day 90 respectively, and the test results are shown in Table 2.

TABLE 2 growth and development of young Cibotium barometz seedlings treated differently

Note that the different lower case letters after the same column of data represent significant differences (p < 0.05)

As can be seen from Table 2, the different seedling substrates have significant effects on the growth and development of the rejuvenation of the young Cinnamomum hirsutum seedlings. The seedling-restoring standard is that the second true leaves grow out, only the T1 group matrix (humic acid soil 1: peat soil 1) reaches the standard within 20 days, the plant height can reach 4.5-5.5 cm within 90 days, the good growth state is shown, and the final survival rate is 79.17%. In contrast, the other three groups have no effective seedling growth, and the growth state is poor, namely the survival rate of the T2 group and the T3 group is respectively reduced to 4.00 percent and 4.33 percent at the 60 th day, the seedlings are all dead within 75 days, and the survival rate of the T4 group at the 75 th day is only 9.67 percent, and the seedlings are all dead within 90 days. In addition, four groups of substrate treatments all had different levels of algae contamination, with the T1 group being less contaminated and the remaining three groups being more contaminated, possibly adversely affecting seedling emergence and growth. The test shows that the substrate of the leaf rot soil (1) is the optimal alloy Mao Gouyou for transplanting and rejuvenating the spore seedlings. In the matrix, young seedlings are subjected to seedling recovery within 20 days, the plant height within 90 days can reach 4.5-5.5 cm, the survival rate is as high as 79.17%, and the matrix is obviously superior to other matrix treatment groups.

5. Water and fertilizer management

And on the 90 th day after the transplanting rejuvenation, a proper amount of nutrient solution is sprayed to continue the culture. The nutrient solution was diluted to a 0.2% strength (mass percent) aqueous solution using a macroelement water-soluble fertilizer (standard NY/T1107-2020) and sprayed once every 7 days, 200ml per pan. Under the culture condition, the illumination intensity is kept at 2500-3500 Lux, the indoor temperature is kept at 23-25 ℃, and the air relative humidity (the air relative humidity in the seedling tray) is kept at 65-85%. After the culture of the nutrition solution is sprayed, the young plant height of the young Cibotium barometz can reach 8.5-10.0 cm, 5-8 leaves can grow on each plant, at the moment, the seedling growth vigor is good, and the subsequent basin-separating seedling hardening can be performed so as to promote the independent growth and the adaptability improvement of the plants. When seedling is planted in a pot, seedlings with the height of 8.5-10.0 cm are planted in the flowerpot in a separated mode, seedling hardening matrixes are selected from leaf rotting soil (1) consistent with the T1 group, peat soil (1) is used for treatment, and the seedling hardening matrixes are optimal matrixes screened in the previous test and are beneficial to promoting healthy growth of the seedlings. And (3) timely watering root fixing water after planting, and placing the root fixing water in a seedling hardening greenhouse, wherein the temperature of the greenhouse is kept at 25-30 ℃. Spraying water mist for 2-3 times every day for 10 minutes each time, so as to ensure that the relative humidity of the culture air is maintained at 60% -65%, the illumination intensity of the hardening seedlings is 3000-3700 lux, and the illumination time is 12-14 hours. After hardening off for about 30 days (150 days from the time of spore seedling transplanting), the plant height can reach 12-15 cm, the number of leaves is increased to 6-9, the growth state is good, and the plant can enter a further cultivation stage.

Claims

1. A method for artificially cultivating Cibotium barometz, comprising the following steps: transplanting Cibotium barometz sporophyte seedlings to be rejuvenated into a matrix for rejuvenation culture; wherein the matrix is a mixture of leaf mold and peat soil in a volume ratio of (1-1.5):1 or 1:1, or a mixture of leaf mold, white vermiculite and garden soil in a volume ratio of 4:1:3.

2. The method according to claim 1, characterized in that: when the first true leaf of the sporophyte seedling grows to 0.3-0.5 cm, transplanting and rejuvenation culture are carried out.

3. The method according to claim 1 or 2, characterized in that: the conditions of the rejuvenation culture include: a temperature of 23-25°C, a light intensity of 2500-3500 Lux, a light duration of 12-14 hours or 14 hours per day, and a relative humidity of 65%-85% in the space where the sporophyte seedlings are located;

and/or, the water content of the substrate in the rejuvenation culture is 60% to 65% or 60%;

And/or, the rejuvenation culture also includes the step of applying a nutrient solution; specifically, the nutrient solution is a water-soluble fertilizer of a large number of elements; further specifically, the nutrient solution is applied on the 90th day after transplanting, and the water-soluble fertilizer of a large number of elements is diluted into an aqueous solution with a concentration of 0.2% (mass percentage concentration), and sprayed once every 7 days, with 200 ml sprayed per 1760 square centimeters each time.

4. The method according to any one of claims 1 to 3, characterized in that: the method further comprises a step of hardening the seedlings, wherein the sporophyte seedlings to be hardened are planted separately, and the matrix is a mixture of leaf mold and peat soil in a volume ratio of (1-1.5):1 or 1:1.

5. The method according to any one of claims 1 to 4, characterized in that: the sporophyte seedlings to be hardened are sporophyte seedlings with a height of 8.5-10.0 cm or sporophyte seedlings with 5 to 8 leaves per plant;

And/or, the conditions for hardening the seedlings include: a temperature of 25-30° C., a relative humidity of 60%-65% in the greenhouse, a light intensity of 3000-3700 lux, and a light duration of 12-14 hours.

6. The method according to any one of claims 1 to 5, characterized in that the method comprises the following steps: sowing spores of Cibotium barometz in a substrate, first subjecting the substrate to dark treatment, and then culturing the substrate to obtain sporophyte seedlings to be rejuvenated.

7. The method according to any one of claims 1-6 is characterized in that: the conditions of the dark treatment include: culturing in darkness for 44 to 48 hours or 48 hours; specifically, the conditions of the dark treatment include: culturing in darkness and at a temperature of 23 to 25°C for 44 to 48 hours or 48 hours; further specifically, the conditions of the dark treatment include: culturing in darkness, at an initial substrate humidity of 60% or 60% to 65%, at a temperature of 23 to 25°C, and in a closed condition for 44 to 48 hours or 48 hours; further specifically, the conditions of the dark treatment include: culturing in darkness, at an initial substrate humidity of 60% or 60% to 65%, at a temperature of 23 to 25°C, and in a closed condition for 44 to 48 hours or 48 hours, without replenishing water during the period.

8. The method according to any one of claims 1 to 7, characterized in that: the step of re-culturing to obtain the sporophyte seedlings to be rejuvenated comprises: culturing until the gametophyte sexual organs mature under the following conditions: the ambient temperature is 23-25° C., the light intensity is 2500-3500 Lux, the light is illuminated for 12-14 hours or 14 hours per day, and the relative humidity of the air in the environment where the gametophyte is located is 65-85%; specifically, the step of re-culturing to obtain the sporophyte seedlings to be rejuvenated comprises: culturing until the gametophyte sexual organs mature under the following conditions: the ambient temperature is 23-25° C., the light intensity is 2500-3500 Lux, the light is illuminated for 12-14 hours or 14 hours per day, and the relative humidity of the air in the environment where the gametophyte is located is 65-85%, and the environment is sealed.

9. The method according to any one of claims 1 to 8, characterized in that after the gametocyte sexual organs mature, the method comprises the following steps:

(1) The relative humidity of the air in the environment where the gametophytes are located is adjusted to 65-85%, and then the gametophytes are cultured in a sealed environment at a constant temperature of 25° C. for 10 days without water replenishment; specifically, the relative humidity of the air in the environment where the gametophytes are located is adjusted to 65-85%, and then the gametophytes are cultured in a sealed environment at a constant temperature of 25° C. with a light intensity of 2500-3500 Lux and 12-14 hours or 14 hours of light per day for 10 days without water replenishment;

(2) the sealed condition is then released, and the gametocytes are placed in a culture medium at 23-25°C, with a light intensity of 2500-3500 Lux, with light exposure for 12-14 hours or 14 hours per day, and a relative humidity of 22-25% for 22-24 hours or 24 hours;

(3) Promote fertilization by spraying water mist.

10. The method according to any one of claims 1 to 9, characterized in that:

The method comprises the following steps: after fertilization is completed, culturing until the sporophyte seedlings germinate; more specifically, after fertilization is completed, culturing until the sporophyte seedlings germinate under the following conditions: an ambient temperature of 23 to 25°C, a light intensity of 2500 to 3500 Lux, 12 to 14 hours or 14 hours of light per day, and an air relative humidity of 65 to 85%; and even more specifically, the culturing is carried out under closed conditions.

and/or, after the sporophyte seedlings germinate, culturing under the following conditions until the sporophyte seedlings to be rejuvenated are obtained: light intensity of 3000-4000 Lux, 12-14 hours or 14 hours of light per day, temperature of 26-27° C., relative air humidity maintained at 55%-70%;

And/or, the substrate for spore inoculation is peat soil, specifically Pinnaceous peat soil, with a particle size of 0 to 6 mm and a pH value of 5.5 to 6.0;

And/or, the initial moisture content of the substrate used for spore inoculation is 60%.