A method for artificially breeding blackheart ferns
By optimizing the propagation substrate and culture conditions of blackheart fern, and combining effective pest and disease management, the problem of low propagation efficiency of blackheart fern has been solved, achieving high-efficiency spore germination and seedling survival, thus meeting the needs of horticulture and industry.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Patents(China)
- Current Assignee / Owner
- SHENZHEN ORCHID PLANT PROTECTION RES CENT
- Filing Date
- 2024-09-14
- Publication Date
- 2026-05-15
AI Technical Summary
Blackheart fern is scarce in the wild, has low breeding efficiency, and is difficult to meet market demand.
A specific ratio of planting organic matter, Pinscher peat nutrient soil and vermiculite mixture is used as the propagation and rejuvenation substrate, combined with high-pressure steam sterilization, reasonable cultivation conditions and pest and disease management methods, including the use of glutaraldehyde and iprodione suspension to treat contamination.
It improves spore germination rate, seedling survival rate and growth rate, is easy to operate, and is suitable for large-scale propagation by horticulture enthusiasts and industrial parks.
Smart Images
Figure CN119183939B_ABST
Abstract
Description
Technical Field
[0001] This invention relates to the field of artificial propagation technology of plants, specifically to a method for artificial propagation of blackheart fern. Background Technology
[0002] Black-hearted fern (Doerryoptis concolor) belongs to the genus Doryopteris in the family Pteridaceae. As a unique and rare ornamental plant, it is beloved by horticultural enthusiasts and attracts the attention of botanists due to its distinctive leaf shape, deep leaf color, and growth characteristics. The entire plant has heat-clearing, diuretic, and hemostatic properties, and is commonly used to treat urinary tract infections and traumatic bleeding. Black-hearted fern is mainly distributed in Guangdong and Hainan provinces, and its endangered status is assessed as Vulnerable (VU). Due to its strict habitat requirements, narrow distribution range, and extremely difficult spore germination in the wild, the wild population of black-hearted fern is even scarcer. Therefore, developing a simple and effective method for spore propagation of black-hearted fern is of great significance for improving its propagation efficiency and promoting the development of its industry. Summary of the Invention
[0003] To solve the aforementioned technical problems, the inventors of this invention, through extensive exploration, developed a technique for the artificial propagation of black-heart ferns. This technique is highly efficient and low-cost, and has significant implications for production practice.
[0004] This invention provides a method for artificially propagating blackheart fern (Doerryoptis concolor), the method comprising the following steps:
[0005] (1) Sow the spores of blackheart fern in the propagation substrate and cultivate them until sporophyte seedlings are obtained; (2) Transplant the sporophyte seedlings into the rejuvenation substrate for division and rejuvenation culture; wherein, the propagation substrate is a mixture of planting organic matter, Pins peat nutrient soil and vermiculite in a 4:1:1 (volume ratio); the rejuvenation substrate is a mixture of planting organic matter, Pins peat nutrient soil and vermiculite in a 4:1:1 (volume ratio).
[0006] Among them, organic matter for planting is organic matter specifically for planting plants. For example, the manufacturer is Jilin Yafeng Fertilizer Co., Ltd., the enterprise standard is Q / YFFL 001-2021, and the product name is organic matter.
[0007] Pins Top Peat Potting Soil: Produced by Pins Top Group of Denmark, it is a general-purpose type with a particle size of 0-10mm.
[0008] The propagation and rejuvenation substrates used in this method were obtained through extensive screening and experience summarization. Compared with other substrates, they not only enable successful propagation, but also have a high spore germination rate, a high survival rate of rejuvenated seedlings, and mild disease.
[0009] In the above method, preferably, the propagation substrate is sterilized; the rejuvenation substrate does not require sterilization. Sterilizing the propagation substrate can greatly reduce or almost eliminate moss contamination. The sterilization method is preferably autoclaving.
[0010] In the above method, the preferred cultivation conditions throughout the entire cultivation process are: a temperature of 23-25℃, a light intensity of 2000 lux, and a daily light duration of 14 hours. Under these conditions, successful propagation can be achieved, and the spore germination rate, gametophyte formation rate, and sporophyte formation rate can be improved.
[0011] The methods described above are generally susceptible to pests and diseases, such as algal and fungal contamination (mainly mold). To address these contaminations, the inventors of this invention have explored effective remediation methods, the preferred methods being as follows:
[0012] For algal contamination, dilute a 20% glutaraldehyde solution at a ratio of 1:300 and add it dropwise to the contaminated area. Preferably, use once daily for 3 consecutive days; the dosage each time is approximately 200 ml per square meter (5.6 trays), such as 180-220 ml or 200 ml.
[0013] For fungal contamination (mainly mold), spray the contaminated area with a mixture of iprodione suspension, chlorothalonil suspension, and water in a ratio of 1:1:1200 (by mass). Preferably, spray once a day for three consecutive days; the dosage per square meter (5.6 trays) is approximately 300 ml, such as 280-320 ml, preferably 300 ml.
[0014] The concentration of iprodione suspension is 500 g / L, and the concentration of chlorothalonil suspension is 40%.
[0015] Preferably, in step (1), when the gametophytes mature, the gametophyte group is sprayed promptly, once every 3 days, with the amount of water used for each spray being enough to make water droplets appear on the surface of the gametophytes, until sporophyte seedlings are obtained. Spraying treatment can promote sperm leakage and motility, enabling sperm and egg to combine and form sporophytes, which can both increase the number of sporophytes formed and accelerate the culture process.
[0016] Under normal circumstances, this method takes about 60 days from sowing to gametophyte maturity; and from gametophyte maturity to obtaining a large number of sporophyte seedlings with uniform emergence, it generally takes 70-80 days.
[0017] Preferably, in the above method, sterile water is sprayed into the breeding substrate to make the initial moisture content of the breeding substrate 25% (by mass); water is sprayed into the rejuvenation substrate to make the initial moisture content of the rejuvenation substrate 25% (by mass).
[0018] Preferably, the propagation substrate is sieved, but the rejuvenation substrate is not sieved. The sieve is a 1mm aperture sieve.
[0019] Preferably, the container used for sowing is a seedling tray. After sowing, the seedling tray is covered with a transparent cover in time, and the seedling tray and the transparent cover are sealed with tape to keep the substrate moist. After the gametophytes mature, the sealing tape between the seedling tray and the transparent cover is removed.
[0020] Preferably, the container for division and rejuvenation is a seedling tray with a transparent lid, and there is no need to seal between the seedling tray and the transparent lid.
[0021] Preferably, the sowing rate is 0.05 ml of spores per square meter (5.6 trays).
[0022] Preferably, in the above method, the method for obtaining the spores is as follows: placing leaves with mature spores but unopened sacs into a paper bag, sealing it, and then placing it in a dry and ventilated place to dry until the spores naturally dry and fall into the paper bag;
[0023] Preferably, when the sporophyte seedlings reach a height of 0.8-1.2 cm (preferably 1 cm), the sporophyte seedlings are transplanted into the rejuvenation substrate.
[0024] The inventors of this invention conducted extensive research and experimentation on culture media and conditions, as well as pest and disease management, ultimately arriving at a method for propagating black-hearted ferns. This method boasts high germination rates, high survival rates, high growth quality, and rapid growth, effectively meeting market demand for black-hearted ferns. Furthermore, the method is simple and easy to implement, requiring no complex equipment or techniques. This invention is not only suitable for home propagation by horticultural enthusiasts but also for large-scale industrial parks dedicated to the propagation of black-hearted ferns, providing technical support for fields such as ornamental horticulture, ecological restoration, and the development of medicinal plant resources, thus possessing significant application value. Attached Figure Description
[0025] Figure 1 It is the sporophyte of the blackheart fern.
[0026] Figure 2 It is a substrate used for sowing.
[0027] Figure 3 These are blackheart fern spores.
[0028] Figure 4 Use electrical tape to seal the seedling tray and transparent cover.
[0029] Figure 5 To cultivate an environment.
[0030] Figure 6 It is a mature gametophyte.
[0031] Figure 7It is a sporophyte seedling.
[0032] Figure 8 These are spore-forming seedlings.
[0033] Figure 9 To rejuvenate the seedlings. Detailed Implementation
[0034] The following are some specific embodiments of the present invention. Unless otherwise specified, the technical means or reagents used are conventional methods and reagents known in the art.
[0035] The leaves of the blackheart fern (Doerryoptis concolor) were collected from rocks beside a stream in the forest of Wutong Mountain, Shenzhen, Guangdong Province.
[0036] Example 1: Artificial propagation of blackheart fern
[0037] Healthy, mature black-heart fern plants were selected from rocks along a stream in the forest of Wutong Mountain. Harvesting was carried out when the spores were mature but the sacs were not yet open. During collection, the leaves containing the spores ( Figure 1 Cut off the spores, put them in a paper bag made of newspaper, seal it, and place it in a dry and ventilated place to dry. After about 10 days, the spores will naturally dry and fall into the bag.
[0038] Mix planting organic matter, Pinscher peat moss, and vermiculite in a 4:1:1 (volume ratio); after mixing, sieve the mixture through a 1mm sieve. Place the sieved soil into heat-resistant inoculum bags, sterilize by high-pressure steam at 125℃ for 25 minutes, remove and let cool. Spread the mixture evenly in a seedling tray measuring 55cm long, 32cm wide, and 8cm deep, with a soil layer thickness of 5cm. Spray the substrate with sterile distilled water to moisten it, bringing the initial substrate moisture content to 25% (by weight). Figure 2 The pH level is natural.
[0039] Pins Top Peat Potting Soil: Produced by Pins Top Group of Denmark, it is a general-purpose type with a particle size of 0-10mm.
[0040] Organic matter for planting: Manufacturer is Jilin Yafeng Fertilizer Co., Ltd., Enterprise Standard: Q / YFFL 001-2021, Trade name is Organic Matter.
[0041] Lay a sheet of A4 white paper flat on the table. Gently shake the dried spore leaves from one corner of the newspaper bag and transfer the spores that have naturally scattered inside the bag onto the white paper. Figure 3 Sow the spores evenly and promptly in the seedling trays at a rate of approximately 0.05 ml per square meter (5.6 trays).
[0042] Immediately after sowing, cover the seedling trays with transparent lids, and seal the gap between the trays and the lids with 18mm wide black electrical tape. Figure 4 To keep the propagation substrate moist, while controlling the culture environment ( Figure 5 The temperature is 23-25℃, the sunlight intensity is 2000 lux, and the daily sunlight duration is 14 hours.
[0043] Gametophytes begin to appear about 30 days after sowing and mature about 60 days later. At this point, the sealing tape is removed, and after another 5-7 days, all gametophytes will be fully developed. Figure 6 After the gametophytes mature, promptly spray the gametophyte group with tap water to promote the release and movement of sperm cells from the antheridium, facilitating fertilization and the formation of sporophytes. Use only a small amount of water per spray, just enough to make tiny water droplets appear on the surface of the gametophytes. Spray every 3 days to keep the area moist until a large number of sporophyte seedlings emerge uniformly. See the image of sporophyte seedlings. Figure 7 When the sporophyte seedlings reach a height of 1 cm, they should be divided and rejuvenated in a timely manner. Figure 8 The substrate for division and rejuvenation is as follows: Mix planting organic matter, Pinscher peat moss, and vermiculite in a 4:1:1 (volume ratio). No sieving or sterilization is required. Spray with tap water to bring the initial substrate moisture content to 25% (by weight) before dividing and replanting. The containers for division and rejuvenation are seedling trays with transparent lids, but sealing with tape is not necessary. Post-division cultivation conditions: temperature 23-25℃, light intensity 2000 lux, and daily light duration 14 hours. Under these conditions, after 65-60 days, each plant will have 5-8 leaves, and the rejuvenated seedlings will reach a height of 8-10 cm. Figure 9 ).
[0044] Minor diseases may occur during cultivation. The treatment methods for various diseases and pests are as follows:
[0045] Algae contamination: Dilute 20% glutaraldehyde solution at a ratio of 1:300 and add it dropwise to the contaminated area. The dosage is approximately 200 ml per square meter (5.6 trays) each time, once a day for three consecutive days.
[0046] A 20% concentration glutaraldehyde solution is marketed as Concentrated Glutaraldehyde Solution (for Aquaculture).
[0047] Moss contamination: Sterilization of the seeding substrate has solved the problem of moss contamination;
[0048] Miscellaneous fungal diseases: mainly caused by molds, including gray mold and powdery mildew. Use a mixture of iprodione suspension, chlorothalonil suspension, and tap water in a ratio of 1:1:1200 (by weight), and spray it onto the contaminated area. The dosage is approximately 300 ml per square meter (5.6 trays) each time, once a day for 3 consecutive days.
[0049] The active ingredient and content of chlorothalonil suspension is 40% chlorothalonil, and the dosage form is suspension. The concentration of isoproporitide suspension is 500g / L.
[0050] Results: The number of sporophyte seedlings produced per square meter (5.6 trays) was approximately 9900, 10500, and 9200, respectively. The survival rate of the rejuvenated sporophyte seedlings after division was 80%, 83%, and 90%, respectively. The cost of each 10cm tall rejuvenated (commercial) seedling was approximately 0.2 yuan. The formula for calculating the survival rate of the rejuvenated seedlings after division was: 1120 seedlings were divided per square meter (5.6 trays). After 20 days, the number of surviving seedlings was counted. The survival rate (%) was calculated as: (Number of surviving seedlings / 1120) × 100%.
[0051] Example 2
[0052] The sowing substrate in Example 1 was replaced with the following: organic matter (sterilized), peat moss + rice husk charcoal (unsterilized). The results are shown in Table 1.
[0053] Table 1
[0054]
[0055] Pins Top Peat Potting Soil: Produced by Pins Top Group of Denmark, it is a general-purpose type with a particle size of 0-10mm.
[0056] Organic matter for planting: Manufacturer is Jilin Yafeng Fertilizer Co., Ltd., Enterprise Standard: Q / YFFL 001-2021, Trade name is Organic Matter.
[0057] Germination rate calculation method: Place defatted cotton in a petri dish, sterilize the leachate of the sowing substrate, and then soak the defatted cotton in it. Use the condition that no water flows out when tilted as a standard. Sow blackheart spores. After 10 days, check the germination status. Select three different fields of view under a stereomicroscope and calculate the spore germination rate for each. The germination rate (%) is calculated as: (Number of germinated spores / Total number of spores in the field of view) × 100%. The average germination rate of the three fields of view is the final germination rate.
[0058] Method for obtaining leachate from sowing substrate: Add twice the volume of tap water to the substrate to be sown, stir thoroughly, let stand for 12 hours, take the supernatant, filter it through three layers of gauze, and use the filtrate for spore germination experiments.
[0059] Example 3
[0060] The division rejuvenation substrate in Example 1 was replaced with different substrates, and the results are shown in Table 2.
[0061] Table 2
[0062]
[0063] As can be seen from Groups A and B in Table 2, Pins peat nutrient soil has a significant effect on the prevention and control of pests and diseases, and can also significantly improve the survival rate of rejuvenated plants.
[0064] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.
Claims
1. An artificially bred blackheart fern ( Doerryoptis concolor The method includes the following steps: (1) Sow the spores of blackheart fern in the propagation substrate and cultivate until sporophyte seedlings are obtained; (2) Transplant sporophyte seedlings into a rejuvenation substrate for division and rejuvenation culture; in, The propagation substrate is a mixture of planting organic matter, Pins peat nutrient soil and vermiculite in a volume ratio of 4:1:1; The rejuvenation substrate is a mixture of planting organic matter, Pins peat moss, and vermiculite in a volume ratio of 4:1:1; The method also includes a step of pest and disease control: For algal contamination, dilute a 20% glutaraldehyde solution at a ratio of 1:300 and add it dropwise to the contaminated area once a day for three consecutive days, with each application being 180ml-220ml per square meter. For fungal contamination, spray the contaminated area with a mixture of iprodione suspension, chlorothalonil suspension and water in a mass ratio of 1:1:1200; wherein the concentration of iprodione suspension is 500g / L and the concentration of chlorothalonil suspension is 40%; spray once a day for 3 consecutive days, with each application being 280ml-320ml per square meter; In step (1), when the gametophytes mature, the gametophyte group is sprayed in a timely manner, once every 3 days. The amount of water used for each spray is enough to make water droplets appear on the surface of the gametophytes until sporophyte seedlings are obtained. Spray sterile water into the propagation substrate to make the initial moisture content of the propagation substrate 25%; the container used for sowing is a seedling tray. After sowing, cover the seedling tray with a transparent cover in time, and seal the seedling tray and the transparent cover with tape to keep the substrate moist; after the gametophytes mature, remove the sealing tape between the seedling tray and the transparent cover. The sowing rate is 0.05 ml of spores per square meter of seedling tray.
2. The method according to claim 1, characterized in that: The breeding substrate is sterilized; the rejuvenation substrate does not require sterilization.
3. The method according to claim 1 or 2, characterized in that: In the method described, the temperature is 23-25℃, the light intensity is 2000 lux, and the daily light exposure time is 14 hours.
4. The method according to any one of claims 1-3, characterized in that: For algal contamination, the dosage is 200ml per square meter each time; for bacterial contamination, the dosage is 300ml per square meter each time.
5. The method according to any one of claims 1-4, characterized in that: The method includes the following steps: spraying water into the rejuvenation matrix to make the initial moisture content of the rejuvenation matrix 25%.
6. The method according to any one of claims 1-5, characterized in that: The breeding substrate is sieved, but the rejuvenation substrate is not sieved.
7. The method according to any one of claims 1-6, characterized in that: The containers for division and rejuvenation are seedling trays and transparent lids, and there is no need to seal between the seedling trays and the transparent lids.
8. The method according to any one of claims 1-7, characterized in that: The method for obtaining the spores is as follows: Place mature leaves with unopened spore sacs into a paper bag, seal the bag, and place it in a dry and ventilated place to dry until the spores naturally dry and fall into the paper bag.
9. The method according to any one of claims 1-8, characterized in that: When the sporophyte seedlings reach a height of 0.8-1.2 cm, transplant them into the rejuvenation substrate.
10. The method according to any one of claims 1-9, characterized in that: When the sporophyte seedlings reach a height of 1 cm, transplant them into the rejuvenation substrate.