Method for artificially breeding black heart fern

Abstract

The invention provides a method for artificially breeding Doerrythropodium concolor (Doerrythropodium concolor). The method comprises the following steps of: selecting a culture medium for breeding the Doerrythropodium concolor; the method for artificially breeding the black heart fern comprises the following steps: (1) sowing spores of the black heart fern in a breeding matrix, and culturing until sporophyte seedlings are obtained; and (2) transplanting the sporophyte seedlings into a rejuvenation matrix, and carrying out division rejuvenation culture. The method has the advantages of high germination rate, high survival rate, high growth quality and high growth speed, and can effectively meet the requirements of the market on the black heart fern.

Description

Technical Field

[0001] The invention relates to the technical field of artificial propagation of plants, and in particular to an artificial propagation method of rudbeckia radiata. Background Art

[0002] Doryopteris concolor belongs to the genus Doryopteris in the family Pteridaceae. As a unique and rare ornamental plant, it is deeply loved by gardening enthusiasts and the attention of botanists because of its unique leaf shape, deep leaf color and growth characteristics. The whole plant of Doryopteris concolor has the functions of clearing heat, diuresis and hemostasis, and is often used for urinary tract infection, traumatic bleeding and other symptoms. Doryopteris concolor is mainly distributed in Guangdong, Hainan and other places, and its endangered level is assessed as vulnerable (VU). Due to the strict habitat requirements of Doryopteris concolor, the narrow distribution area, and the extremely difficult germination of spores in the wild, the number of wild resources of Doryopteris concolor is even scarcer. Therefore, the development of a simple and effective method for breeding spores of Doryopteris concolor is of great significance to improving the breeding efficiency of Doryopteris concolor and promoting its industrial development. Summary of the invention

[0003] In order to solve the above technical problems, the inventor of the present invention has invented a set of artificial breeding technology of black heart fern after a lot of exploration. The technology is high in efficiency and low in cost, and has important significance in production practice.

[0004] The present invention provides a method for artificially breeding Doerryoptis concolor, which comprises the following steps:

[0005] (1) sowing spores of black fern in a breeding matrix and culturing until sporophyte seedlings are obtained; (2) transplanting the sporophyte seedlings into a rejuvenation matrix for division and rejuvenation culture; wherein the breeding matrix is ​​a mixture of organic matter for planting, Pin's peat nutrient soil and vermiculite in a volume ratio of 4:1:1; and the rejuvenation matrix is ​​a mixture of organic matter for planting, Pin's peat nutrient soil and vermiculite in a volume ratio of 4:1:1.

[0006] Among them, organic matter for planting is organic matter specially used 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] Pin's peat nutrient soil: The manufacturer is Denmark's Pin's Top Group, the specification model is general type, and the particle size is 0-10mm.

[0008] The breeding matrix and rejuvenation matrix in this method are obtained through a large amount of screening and experience summary. Compared with other matrices, they can not only achieve successful breeding, but also have a high spore germination rate, a high survival rate of rejuvenation seedlings, and mild diseases.

[0009] In the above method, preferably, the breeding substrate is sterilized; the rejuvenation substrate does not need to be sterilized. Sterilizing the breeding substrate can greatly reduce or almost eliminate moss contamination. The sterilization method can preferably be high-pressure steam sterilization.

[0010] In the above method, during the whole culture process, the preferred culture conditions are: temperature of 23-25°C, light intensity of 2000 lux, and illumination time of 14 hours per day. Under these conditions, successful breeding can be achieved, and the spore germination rate, gametophyte and sporophyte formation rate can also be improved.

[0011] In the above methods, pests and diseases will generally exist, such as algae pollution and fungal pollution (mainly mold). In response to these pollutions, the inventors of the present invention have explored effective treatment methods, preferably as follows:

[0012] For algae pollution, a 20% glutaraldehyde solution is diluted at 1:300 and then dripped onto the polluted area. Preferably, once a day for 3 consecutive days; each dosage is about 200 ml per square meter (5.6 trays), such as 180-220 ml, or 200 ml.

[0013] For fungal contamination (mainly mold), a mixture of 1:1:1200 (mass ratio) of iprodione suspension, thiophanate-methyl suspension and water is sprayed on the contaminated area. Preferably, spray once a day for 3 consecutive days; the dosage per square meter (5.6 trays) is about 300 ml, such as 280-320 ml, preferably 300 ml.

[0014] The concentration of the iprodione suspension concentrate was 500 g / L, and the concentration of the thiophanate-methyl suspension concentrate was 40%.

[0015] Preferably, in step (1), when the gametophytes mature, the gametophyte group is sprayed in time, once every 3 days, and the amount of water used for each spray is until water droplets appear on the surface of the gametophytes, until sporophyte seedlings are obtained. Spraying treatment can promote sperm overflow and swimming, achieve sperm-egg combination and then form sporophytes, which can not only increase the number of sporophytes formed, but also accelerate the culture process.

[0016] Generally speaking, this method takes about 60 days from sowing to gametophyte maturity, and 70-80 days from gametophyte maturity to obtaining a large number of sporophyte seedlings and their emergence is uniform.

[0017] Preferably, in the above method, sterile water is sprayed into the breeding matrix so that the water content of the initial breeding matrix is ​​25% (mass percentage); water is sprayed into the rejuvenation matrix so that the water content of the initial rejuvenation matrix is ​​25% (mass percentage).

[0018] Preferably, the breeding matrix is ​​sieved, and the rejuvenation matrix is ​​not sieved, wherein the sieve has a 1 mm aperture.

[0019] Preferably, the container for sowing is a seedling tray. After sowing, the seedling tray is promptly covered with a transparent cover, and the seedling tray and the transparent cover are sealed with tape to keep the substrate moist. After the gametophyte matures, 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 and a transparent cover, and there is no need to seal between the seedling tray and the transparent cover.

[0021] Preferably, the sowing rate is 0.05 ml of spores per square meter (5.6 trays).

[0022] Preferably, in the above method, the spores are obtained as follows: leaves with mature spores but unopened sacs are placed in a paper bag, sealed and placed in a dry and ventilated place to dry until the spores dry naturally and fall off in 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 a rejuvenation medium.

[0024] The inventors of the present invention have conducted a lot of explorations on culture substrates and culture conditions, as well as experimental explorations on pest and disease management, and finally obtained a method for breeding blackheart fern. The method of the present invention has a high germination rate, a high survival rate, a high growth quality, and a fast growth rate, and can effectively meet the market demand for blackheart fern. Moreover, the method of the present invention is simple and easy to operate, and does not require complex equipment and technology. The present invention is not only suitable for home breeding of gardening enthusiasts, but can also be used in industrial parks for large-scale breeding of blackheart fern, providing technical support for the fields of gardening appreciation, ecological restoration, and medicinal plant resource development, and has important application value. BRIEF DESCRIPTION OF THE DRAWINGS

[0025] Figure 1 It is the spore leaf of blackheart fern.

[0026] Figure 2 A substrate for sowing.

[0027] Figure 3 It is the spores of black fern.

[0028] Figure 4 Seal the seedling tray and clear lid with electrical tape.

[0029] Figure 5 To cultivate the environment.

[0030] Figure 6 For mature gametophytes.

[0031] Figure 7For sporophyte seedlings.

[0032] Figure 8 It is a spore seedling.

[0033] Fig. 9 To rejuvenate the seedlings. DETAILED DESCRIPTION

[0034] The following are some specific embodiments of the present invention, wherein the technical means or reagents used are conventional methods and reagents known in the art unless otherwise specified.

[0035] The leaves of Doerryoptis concolor were collected from rocks beside a stream in the forest of Wutong Mountain, Shenzhen City, Guangdong Province.

[0036] Embodiment 1 Artificial propagation of black heart fern

[0037] Select healthy and mature black fern plants on rocks beside a stream in the Wutong Mountain forest, and pick them when the spores are mature but the capsules have not yet opened. Figure 1 ) and put them in a paper bag made of newspaper. After sealing, place them in a dry and ventilated place to dry. After about 10 days, the spores will naturally dry and fall off in the bag.

[0038] Organic matter for planting, Pin's peat nutrient soil and vermiculite were mixed at a ratio of 4:1:1 (volume ratio); after mixing, the mixture was sieved with a sieve aperture of 1 mm. The sieved fine soil was placed in a high temperature resistant bacterial bag, sterilized with high pressure steam at 125°C for 25 minutes, taken out and cooled, and spread in a seedling tray with a length of 55 cm, a width of 32 cm, and a depth of 8 cm. The soil layer was 5 cm thick, and sterile distilled water was sprayed to moisten the substrate so that the initial substrate moisture content was 25% (mass percentage) ( Figure 2 ), pH natural.

[0039] Pin's peat nutrient soil: The manufacturer is Denmark's Pin's Top Group, the specification model is general type, and the particle size is 0-10mm.

[0040] Organic matter for planting: The manufacturer is Jilin Yafeng Fertilizer Co., Ltd., the enterprise standard is Q / YFFL 001-2021, and the product name is organic matter.

[0041] Take an A4 size white copy paper and lay it flat on the table. Gently shake the dried spore leaves from the corner of the newspaper bag to get the spores that have naturally scattered in the paper bag onto the white paper ( Figure 3 ). About 0.05 ml of spores are needed per square meter (5.6 trays) of seedling tray area, and they should be spread in the seedling tray in a timely and even manner.

[0042] After sowing, cover the seedling tray with a transparent lid in time, and seal the gap between the tray and the lid with 18mm wide black electrical tape ( Figure 4 ) to keep the culture medium moist and control the culture environment ( Figure 5 ) is set at 23-25℃, sunlight is 2000lux, and the daily illumination time is 14h.

[0043] From the beginning of sowing, gametophytes begin to appear in about 30 days, and gametophytes mature in about 60 days. At this time, remove the sealing tape. After another 5-7 days, all gametophytes will mature. Figure 6 ). After the gametophyte matures, spray the gametophyte group with tap water in time to promote the overflow and movement of sperm cells in the antheridia, so as to achieve the combination of sperm and egg and form sporophyte. The amount of water used for each spray should not be too much, and it should be sprayed until tiny water droplets appear on the surface of the gametophyte. Spray once every 3 days and keep it moist until the sporophyte seedlings appear in large numbers and emerge neatly. See the picture of sporophyte seedlings. Figure 7 When the sporophyte seedlings reach 1 cm in height, timely division and rejuvenation ( Figure 8 ). The matrix for rejuvenation of ramets is: mix planting organic matter, Pin's peat nutrient soil and vermiculite in a ratio of 4:1:1 (volume ratio). No need to sieve and sterilize. Spray tap water to make the initial matrix moisture content 25% (mass percentage) before transplanting. The container for ramet rejuvenation is a seedling tray and a transparent cover, but no tape is needed to seal it. The culture conditions after ramets: the temperature is 23-25℃, the sunlight is 2000lux, and the daily lighting time is 14h. After 65-60 days under the above conditions, each plant has 5-8 leaves and the rejuvenated seedlings can be as high as 8-10cm ( Fig. 9 ).

[0044] Minor diseases may occur during the cultivation period. The treatment methods for various diseases and pests are as follows:

[0045] Algae pollution: Dilute 20% glutaraldehyde solution at 1:300 and add it to the polluted area. The dosage is about 200 ml per square meter (5.6 trays) once a day for three consecutive days.

[0046] The product name of 20% concentration glutaraldehyde solution is concentrated glutaraldehyde solution (for aquaculture).

[0047] Moss contamination: Sterilization of the seeding substrate has solved the problem of moss contamination;

[0048] Fungal diseases: mainly fungi, mainly gray mold, powdery mildew, etc. Mix chlorpyrifos suspension: chlorothalonil suspension: tap water = 1:1:1200 (mass ratio), spray to the contaminated area, each dosage is about 300ml per square meter (5.6 trays), once a day, and spray continuously for 3 days.

[0049] The active ingredient and content of the thiophanate-methyl suspension is 40% thiophanate-methyl, the dosage form is a suspension, and the concentration of the isoprenaline suspension is 500g / L.

[0050] Results: The number of sporophyte seedlings per square meter (5.6 trays) was about 9900, 10500, and 9200, and the sporophyte seedling division rejuvenation survival rate was 80%, 83%, and 90%; the cost of ten-centimeter-high rejuvenation (commercial) seedlings was about 0.2 yuan per plant. The formula for calculating the division rejuvenation survival rate was: 1120 plants were divided per square meter (5.6 trays), and after 20 days, the number of surviving plants was counted, and the survival rate (%) was calculated as: number of surviving plants / 1120×100%.

[0051] Example 2

[0052] The sowing substrate in Example 1 was replaced with the following: organic matter (sterilized), peat soil + rice husk charcoal (unsterilized). The results are shown in Table 1.

[0053] Table 1

[0054]

[0055] Pin's peat nutrient soil: The manufacturer is Denmark's Pin's Top Group, the specification model is general type, and the particle size is 0-10mm.

[0056] Organic matter for planting: The manufacturer is Jilin Yafeng Fertilizer Co., Ltd., the enterprise standard is Q / YFFL 001-2021, and the product name is organic matter.

[0057] Statistical method for germination rate: Put absorbent cotton in a culture dish, sterilize the extract of the sowing matrix, soak the absorbent cotton, and sow the spores of black heart fern with the standard of no water flowing out when tilted. After 10 days, check the germination situation, select three different fields of view under a stereoscope, and calculate the spore germination rate respectively. The calculation formula of germination rate (%) is: number of germination / total number of spores in the field of view × 100%, and the average of the germination rates of the three fields of view is the final germination rate.

[0058] Method for obtaining the extract of the sowing matrix: add 2 times the volume of tap water to the matrix to be sown, stir well, let it stand for 12 hours, take the supernatant, and filter it with three layers of gauze. The filtrate is used for spore germination experiments.

[0059] Example 3

[0060] The ramet rejuvenation matrix in Example 1 was replaced with different matrices. The results are shown in Table 2.

[0061] Table 2

[0062]

[0063] It can be seen from Group A and Group B in Table 2 that Pin's peat nutrient soil has obvious disease and pest control effects and can also significantly improve the rejuvenation survival rate.

[0064] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A method for artificially breeding Doerryoptis concolor, comprising the following steps: (1) sowing spores of Pteris cretica in a breeding medium and culturing until sporophyte seedlings are obtained; (2) transplanting the sporophyte seedlings into a rejuvenation medium for division and rejuvenation culture; in, The propagation medium is a mixture of organic matter for planting, Pin's peat nutrient soil and vermiculite in a volume ratio of 4:1:1; The rejuvenation medium is a mixture of organic matter for planting, Pin's peat soil and vermiculite in a volume ratio of 4:1:

1.

2. The method according to claim 1, characterized in that: The breeding matrix is ​​sterilized; the rejuvenation matrix does not need to be sterilized.

3. The method according to claim 1 or 2, characterized in that: In the method, the temperature is 23-25° C., the light intensity is 2000 lux, and the illumination time is 14 h per day.

4. The method according to any one of claims 1 to 3, characterized in that: The method also includes the step of pest control: For algae contamination, dilute 20% glutaraldehyde solution at 1:300 and add it dropwise to the contaminated area; For fungal contamination, spray the mixed solution obtained by mixing thiophanate-methyl suspension, benomyl suspension and water in a mass ratio of 1:1:1200 onto the contaminated area; among which, the concentration of thiophanate-methyl suspension is 500g / L, and the concentration of benomyl suspension is 40%.

5. The method according to any one of claims 1 to 4, characterized in that: In step (1), when the gametophytes mature, the gametophyte group is sprayed in time, once every 3 days, and the amount of water used for each spray is until water droplets appear on the surface of the gametophytes, until sporophyte seedlings are obtained; And / or, for algae pollution, the dripping amount is once a day for 3 consecutive days, and the dosage each time is 180ml-220ml per square meter, preferably 200ml; And / or, for fungal contamination, the spraying amount is once a day for 3 consecutive days, and the amount used each time is 280ml-320ml per square meter, preferably 300ml.

6. The method according to any one of claims 1 to 5, characterized in that: The method comprises the following steps: spraying sterile water into the breeding matrix to make the water content of the initial breeding matrix 25%; spraying water into the rejuvenation matrix to make the water content of the initial rejuvenation matrix 25%.

7. The method according to any one of claims 1 to 6, characterized in that: The breeding matrix is ​​sieved, but the rejuvenation matrix is ​​not sieved.

8. The method according to any one of claims 1 to 7, characterized in that: The container used for sowing is a seedling tray. After sowing, the seedling tray is promptly covered with a transparent cover, and the seedling tray and the transparent cover are sealed with tape to keep the substrate moist; after the gametophyte matures, the sealing tape between the seedling tray and the transparent cover is removed; The container for division and rejuvenation is a seedling tray and a transparent cover, and there is no need to seal between the seedling tray and the transparent cover.

9. The method according to any one of claims 1 to 8, characterized in that: The sowing amount was 0.05 ml of spores per square meter of seedling tray.

10. The method according to any one of claims 1 to 9, characterized in that: The method for obtaining the spores is as follows: putting leaves with mature spores but unopened sacs into a paper bag, sealing the bag and placing it in a dry and ventilated place to dry until the spores dry naturally and fall off in the paper bag; When the sporophyte seedlings reach a height of 0.8-1.2 cm, transplant them into rejuvenation substrate, preferably 1 cm.

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