Method for large-scale seedling culture of rhizoma drynariae

Abstract

The invention relates to a large-scale seedling raising method for rhizoma drynariae. The method comprises the steps of rhizoma drynariae greenhouse transplanting, seedling raising pretreatment, rhizoma drynariae spore collection, spore germination, rhizome formation promotion, conventional seedling management and the like, and a large number of seedlings can be obtained. According to the method, fortune's drynaria rhizome spores are promoted to germinate on a camphor tree sawdust substrate through weak illumination and a nutrient solution, and the problems that fortune's drynaria rhizome spores do not germinate due to water shortage and cannot form seedlings due to nutrition deficiency are effectively avoided The growth environment of the rhizoma drynariae cultivated in a greenhouse is fully utilized, and the seedling culture space is saved and the seedling culture cost is greatly reduced by modifying the seedling culture substrate and directly breeding spores; by means of an existing hanging spraying facility, water and nutrient supply for seedling raising is guaranteed, the labor cost is reduced, the spore germination rate and the seedling survival rate are greatly increased, and after small rhizomes are directly adopted for transplantation in subsequent management, the spore collection and sowing difficulty is reduced, and the seedling cultivation time is effectively shortened.

Description

Technical Field

[0001] The present invention specifically relates to a method for large-scale seedling cultivation of Drynaria fortunei, belonging to the technical field of medicinal plant seedling cultivation. Background Art

[0002] Drynaria fortunei, also known as Drynaria baronii, Drynaria quercifolia, Drynaria propinqua, Rhizoma Drynariae, Rhizoma Pyrrosiae, Flying Squirrel, etc., is a commonly used traditional Chinese medicine in clinical practice. It is the rhizome of the perennial epiphytic fern Drynaria roosii or Drynaria baronii of the Polypodiaceae family. First recorded in "Supplementary Materia Medica" in the Tang Dynasty, it has the effects of treating injuries and relieving pain, tonifying the kidney and strengthening bones, and is a commonly used drug for diseases such as rheumatism and osteoarthropathy. In recent years, due to blind excavation, its wild resources have been gradually exhausted, and the ecological environment has been severely damaged. At the same time, with the enhancement of people's health awareness and the increase in the amount of medicine used, therefore, the artificial cultivation of Drynaria fortunei is also imperative.

[0003] There are two methods for the propagation of Drynaria fortunei, namely rhizome division propagation and spore propagation, with rhizome division propagation being the main method. The best planting time for rhizome division propagation is May and June. The main method is to separate the mother plant, with each section being about four or five centimeters, and place it in a shady place. After sprouting, plant it. Spore propagation is to collect the mature spores of Drynaria fortunei and let the spores grow and reproduce. However, the germination rate of spores is low, and scientific spore treatment is required. Finally, for rhizome propagation, only thick and plump rhizomes with buds need to be selected, then cut to about fifteen or sixteen centimeters, and then planted according to a row spacing of forty-five centimeters and a plant spacing of forty centimeters.

[0004] Patent Publication No. CN105660415A provides a method for in vitro preservation of Drynaria fortunei. The specific steps are as follows: Take mature Drynaria fortunei as explants, soak them in 75% ethanol for 30 - 45 s, rinse with sterile water 2 - 3 times, immerse them in 0.1% mercuric chloride with 2 drops of Tween for 4 min, then rinse with sterile water 2 - 3 times again, immerse them in 0.1% mercuric chloride with 2 drops of Tween for 4 min once more, and finally rinse with sterile water 2 - 3 times and cut into pieces 0.5 - 1.0 cm long and inoculate them on the spore germination induction medium. Culture for 10 - 14 days, and greenish - yellow spots can be seen in spore germination, which gradually become more and more and turn green. A large number of prothalli appear after 30 - 40 days, and a large number of sporophytes appear after 60 - 70 days, clustering and showing emerald green. After 80 - 90 days of culture, the sporophytes grow into bud seedlings with 2 needles and a height of about 2 cm. Take down the bud seedlings and inoculate them on the propagation and rooting medium for culture. After 30 - 45 days, the multiplication coefficient of sterile seedlings is 4 - 7, the plant height is 4.0 cm - 6.0 cm, the average number of roots is more than 4, and the average root length is more than 4 cm. Inoculate the test - tube seedlings with 4 - 6 leaves obtained by the propagation and rooting of sporophytes onto the preservation medium for in vitro preservation. Although this method can improve the propagation speed of seedlings, the production cost of seedlings is relatively high (currently, it takes more than 5 years to grow Drynaria fortunei, the output per mu is only thousands of kilograms, and the total value is only 3000 - 4000 yuan. The cost of tissue - cultured seedlings is generally about 1 yuan per plant, and about 5000 plants are needed, and the seedling cost exceeds 5000 yuan). It will be very difficult to obtain good economic benefits by planting tissue - cultured seedlings.

[0005] Patent Publication No. CN111727834A provides a method for intensive breeding of Drynaria fortunei seedlings, which includes the following steps: Mix sphagnum moss, vermiculite and fine sand according to a volume ratio of 10:(1 - 3):(1 - 5), add water to make the water content of the substrate 40 - 50%, sterilize to obtain the mixed substrate, and load it into a plug tray for later use; Soak Drynaria fortunei spores in water in the dark for 48 - 72 hours to prepare a spore suspension, at a rate of 200 - 800 grains / cm 2 ; Place the sown plug tray on a greenhouse tidal seedbed for culture. After the gametophytes are mature, soak the substrate thoroughly with water once. Among them, the culture conditions are: Cover the material above the seedbed that is ventilated, light - transmissive and not easy to condense water droplets; Keep the substrate moist; Control the relative air humidity at 80 - 100%, the lowest environmental temperature at 12℃, and the highest environmental temperature at 26℃; Preferably, control the temperature difference between day and night at more than 10℃. Although this method can improve the germination rate of Drynaria fortunei spores, it increases the workload and difficulty of collecting spores and sowing, and the survival rate of later seedling separation and transplantation will also be greatly affected.

[0006] The patent publication number CN115104518A provides a method for rapid seedling raising of medicinal Drynaria fortunei, which includes the following steps: Harvest mature Drynaria fortunei spore leaves in early spring or autumn and winter every year, lay them flat on a large piece of kraft paper, place them in a ventilated, cool and dry place. After the spores fall off by themselves, collect the spores, pass them through a 250-mesh fine sieve, then put them into a plastic centrifuge tube, and seal and store them in a refrigerator at 0-4°C; Select single Sphagnum papillosum as the sowing substrate, spray the substrate with a 1000-fold solution of 60-80% wettable thiophanate-methyl powder plus a 1000-fold solution of 40-60% soluble cyromazine powder and stir evenly, put them into plastic bags or stack and cover and seal for 40-56h; Select a plastic white No. 2 shallow pot with a length of 465mm, a width of 335mm and a height of 85mm as the sowing container, evenly punch 96 holes with a 2-6mm drill bit at the bottom, wash them with clean water and drain the water for later use; Put the substrate into the shallow pot, knead it into small pieces, level it and press it properly to make the substrate maintain a thickness of 2-4cm, soak it thoroughly with water, take it out and place it on a tray for later use; Count the number of sowing pots required for this time 1-3 days in advance, weigh the total amount of spores, add pure water to a manual pressure sprayer, shake it well, let it stand in the dark and soak for 40-56h, shake it before sowing to form a uniform suspension and spray it on the surface of the substrate; After sowing, place the shallow pot on a culture rack, cover it with a glass plate or a transparent acrylic plate; 7-10 days after sowing, the spores begin to germinate, and no water needs to be added within 25-35 days. Spray a 3000-fold solution of Florel 1 compound fertilizer nutrient solution once 25-35 days after the spores germinate; 45-55 days after the spores germinate, soak the gametophyte thoroughly with water once to form sporophyte seedlings; When the first true leaf of the sporophyte seedlings grows to 3-5mm, it is necessary to separate and transplant the seedlings in time. Use a pipette tip forceps to pick up the cluster of seedlings from the base, and plant them according to a plant spacing of 1-2cm. Spray thoroughly with water, cover with a glass plate and place it under the same sowing conditions for continued cultivation. After 3-5 days of slow seedling growth, spray a 3000-fold solution of Florel 1 compound fertilizer once every 12-18 days. When the seedling height is 2-4cm, divide each cluster of seedlings into two and plant them in a 50-hole seedling tray and move them to a cultivation greenhouse for centralized management. This method is similar to the solution provided by CN111727834A. If implemented, there will be the same problems.

[0007] The existing methods of ramet propagation for seedling raising have a high transplanting survival rate and relatively fast growth rate. However, the ramet method requires a large amount of fresh rhizomes of Drynaria fortunei, making large-scale cultivation impossible. Although the direct seeding method using spore propagation can collect a large number of spores, the spore germination rate is relatively low (not exceeding 20%), the seedling formation period is long, the transplanting survival rate is low, and it is also difficult to achieve large-scale seedling raising. When using tissue culture technology for seedling raising, the cultivation process is cumbersome, greatly increasing the production cost of tissue culture seedlings. It is difficult for growers to accept, and it is also difficult to improve the economic benefits of farmers. So far, there has been no report at home and abroad on directly culturing Drynaria fortunei spores into seedlings under simulated ecological environmental conditions. Through a large number of experiments, we finally explored a set of mature methods and successfully achieved direct seeding of Drynaria fortunei spores for seedling raising, which can quickly cultivate a large number of seedlings and provide the possibility for large-scale artificial cultivation of Drynaria fortunei. Summary of the Invention

[0008] Aiming at the problems of low direct seeding germination rate, long seedling formation period, low transplanting survival rate of Drynaria fortunei spores, high tissue culture investment, and high seedling cost, the purpose of the present invention is to provide a method for quickly propagating seedlings with low requirements for the cultivation environment, low investment, and high spore germination rate, achieving the advantages of being suitable for large-scale production, good formula effect, high seedling formation rate, short cultivation time, strong plant growth, high operability, and high application value.

[0009] The purpose of the present invention is to provide a method for large-scale seedling raising of Drynaria fortunei, and the specific steps are as follows:

[0010] (1) Transplanting Drynaria fortunei in the greenhouse: Build a frame more than 10 cm above the ground in the greenhouse, and then evenly spread camphor sawdust on it with a thickness of 8 - 20 cm as the greenhouse substrate. Collect the rhizomes (with leaves) of Drynaria fortunei that have grown for many years and transplant them into the greenhouse substrate. Use a shading net to reduce the light intensity to 30%, and continuously spray water to make the relative air humidity reach more than 75%.

[0011] (2) Pretreatment before seedling raising: When the leaves of Drynaria fortunei to be cultivated start to change color, evenly spread camphor sawdust on the greenhouse substrate with a thickness of 4 - 8 cm.

[0012] (3) Collecting Drynaria fortunei spores: After the Drynaria fortunei spores (turning brown) are mature, use tweezers or a small wooden stick to tap the Drynaria fortunei leaves to make the spores fall as evenly as possible (not in piles) on the camphor sawdust.

[0013] (4) Spore germination: Use a shading net to reduce the light intensity to below 15% again. During the day, start spraying germination nutrient solution with a hanging spray, spraying for 3 - 5 s every 1 h (to ensure that the surface of the camphor sawdust is wet), and the relative air humidity in the greenhouse reaches more than 100%. Cultivate for 20 - 35 days.

[0014] (5) Promote rhizome formation: 10 days after the spores start to germinate, start spraying the rhizome-promoting nutrient solution using a hanging spray during the day, spraying for 3 - 5 seconds every 1 hour. Remove part of the shading net, restore the light intensity to 30%, and keep the relative humidity in the greenhouse at 80 - 70%. Cultivate for 20 - 35 days;

[0015] (6) Conventional management of seedlings: After the height of the seedlings generally reaches 2 cm, stop spraying the rhizome-promoting nutrient solution, and spray foliar fertilizer (potassium dihydrogen phosphate) to promote the growth of seedlings. Spray at a rate of 0.1 - 1.0 g of foliar fertilizer per square meter of the planting area. Keep the relative humidity in the greenhouse at about 65%. After cultivating seedlings with small rhizomes, take them out for transplantation. 2

[0016] In a specific embodiment, the composition of the germination nutrient solution in step (4) is an aqueous solution containing 0.4 - 1.2 g / L of potassium nitrate, 0.1 - 0.4 g / L of potassium dihydrogen phosphate, 50 - 60 mg / L of inositol, 5 - 10 mg / L of glycine, 0.5 - 1 mg / L of nicotinic acid, 0.01 - 0.1 mg / L of pyridoxine hydrochloride, and 25 - 50 g / L of thiophanate-methyl, with a pH value of about 5.5.

[0017] In a specific embodiment, the composition of the rhizome-promoting nutrient solution in step (5) is an aqueous solution containing 0.06 - 0.15 g / L of naphthylacetic acid, 0.4 - 1.2 g / L of potassium nitrate, 0.1 - 0.4 g / L of potassium dihydrogen phosphate, 50 - 60 mg / L of inositol, 5 - 10 mg / L of glycine, 0.5 - 1 mg / L of nicotinic acid, 0.01 - 0.1 mg / L of pyridoxine hydrochloride, and 25 - 50 g / L of thiophanate-methyl, with a pH value of about 5.5.

[0018] Technical effects

[0019] 1. Promote the germination of Drynaria fortunei spores on the camphor sawdust substrate through weak light and nutrient solution, effectively avoiding problems such as the non-germination of Drynaria fortunei spores due to water shortage and the inability to form seedlings due to lack of nutrition;

[0020] 2. Make full use of the growth environment of Drynaria fortunei cultivated in the greenhouse, directly transform the seedling-raising substrate, and use spores for breeding directly, saving seedling-raising space and greatly reducing the seedling-raising cost;

[0021] 3. With the existing hanging spray facilities, this invention ensures the supply of water and nutrients for seedling raising, reduces labor costs, greatly improves the spore germination rate and seedling survival rate. After transplantation with small rhizomes in subsequent management, it reduces the difficulty of spore collection and uniform sowing, and can effectively shorten the seedling cultivation time. Brief description of the drawings

[0022] Figure 1 : Site map of Drynaria fortunei cultivation in the greenhouse;

[0023] Figure 2 : Field picture of Drynaria fortunei spores before maturity in the greenhouse;

[0024] Figure 3 : Schematic diagram of Drynaria fortunei spores after germination in the greenhouse;

[0025] Figure 4 : Schematic diagram of Drynaria fortunei seedlings before transplanting;

[0026] Figure 5 : Schematic diagram of spraying nutrient solution onto camphor wood sawdust after spore germination;

[0027] Figure 6 : Schematic diagram of spraying water onto camphor wood sawdust after spore germination. Detailed implementation method

[0028] Next, the present invention will be further illustrated with examples, but it is not limited to any one of these examples or similar instances.

[0029] Example 1

[0030] Build a frame more than 10 cm above the ground in the greenhouse, and then evenly spread camphor wood sawdust on it with a thickness of 8 - 20 cm as the greenhouse substrate. Collect the rhizomes (with leaves) of Drynaria fortunei that have grown for many years and transplant them into the greenhouse substrate (see Figure 1 ). Use a shading net to reduce the light intensity to 30%, and continuously spray water to make the relative air humidity reach more than 75%; when the leaves of the cultivated Drynaria fortunei start to change color (age), evenly sprinkle camphor wood sawdust on the greenhouse substrate with a thickness of 4 - 8 cm (see Figure 2 ); after the Drynaria fortunei spores (turning brown) mature, use tweezers or a small wooden stick to tap the Drynaria fortunei leaves to make the spores scatter as evenly as possible (not in piles) on the camphor wood sawdust; use the shading net to reduce the light intensity to below 15% again, and start spraying germination nutrient solution by hanging spray during the day, spraying for 3 - 5 s every 1 h (to ensure that the surface of the camphor wood sawdust is moist and no water can be squeezed out by hand), and the relative air humidity in the greenhouse reaches more than 100%, and cultivate for 20 - 35 days; 10 days after the spores start to germinate (see Figure 3 ), start spraying rhizome-promoting nutrient solution by hanging spray during the day, spraying for 3 - 5 s every 1 h, remove part of the shading net, restore the light intensity to 30%, and the relative air humidity in the greenhouse reaches 80 - 70%, and cultivate for 20 - 35 days; after the height of the seedlings generally reaches 2 cm, stop spraying the rhizome-promoting nutrient solution, and use foliar fertilizer (potassium dihydrogen phosphate) to promote the growth of the seedlings, spraying according to 0.1 - 1.0 g of foliar fertilizer / m 2 for spraying, and the relative air humidity in the greenhouse reaches about 65%, and cultivate into seedlings with small rhizomes (see Figure 4 ), then take them out for transplanting, and the transplanting survival rate reaches 98%.

[0031] Control Example 1

[0032] Build a frame more than 10 cm above the ground in the greenhouse, then evenly spread camphor sawdust on it with a thickness of 8 - 20 cm as the greenhouse substrate. Collect the rhizomes (with leaves) of Drynaria fortunei that have grown for many years and transplant them into the greenhouse substrate. Use a shading net to reduce the light intensity to 30%. Continuously spray water to make the relative air humidity reach over 75%. After the spores of Drynaria fortunei (turning brown) mature, use tweezers or a small wooden stick to tap the leaves of Drynaria fortunei to make its spores scatter as evenly as possible (not in piles) on the camphor sawdust. Use the shading net to reduce the light intensity to below 15% again. During the day, start spraying germination nutrient solution by hanging spray, spraying for 3 - 5 s every 1 h, and make the relative air humidity in the greenhouse reach over 100%. Cultivate for 20 - 35 days. After 10 days when the spores start to germinate (see Figure 5 , the spore germination rate is relatively low), during the day, start spraying rhizome-promoting nutrient solution by hanging spray, spraying for 3 - 5 s every 1 h. Remove part of the shading net and restore the light intensity to 30%. Make the relative air humidity in the greenhouse reach 80 - 70%. Cultivate for 20 - 35 days. After the height of the seedlings generally reaches 2 cm, stop spraying the rhizome-promoting nutrient solution and use foliar fertilizer (potassium dihydrogen phosphate) to promote the growth of the seedlings. Spray according to 0.1 - 1.0 g of foliar fertilizer / m 2 for spraying. Make the relative air humidity in the greenhouse reach about 65%. After cultivating into seedlings with small rhizomes, take them out for transplanting, and the transplanting survival rate reaches 35%.

[0033] Control Example 2

[0034] Build a frame more than 10 cm above the ground in the greenhouse, then evenly spread camphor sawdust on it with a thickness of 8 - 20 cm as the greenhouse substrate. Collect the rhizomes (with leaves) of Drynaria fortunei that have grown for many years and transplant them into the greenhouse substrate. Use a shading net to reduce the light intensity to 30%. Continuously spray water to make the relative air humidity reach over 75%. After the spores of Drynaria fortunei (turning brown) mature, use tweezers or a small wooden stick to tap the leaves of Drynaria fortunei to make its spores scatter as evenly as possible (not in piles) on the camphor sawdust. Use the shading net to reduce the light intensity to below 15% again. During the day, start spraying water by hanging spray, spraying for 3 - 5 s every 1 h, and make the relative air humidity in the greenhouse reach over 100%. Cultivate for 20 - 35 days. After 10 days when the spores start to germinate (see Figure 6 , the spore germination rate is very low), during the day, continue to start spraying water by hanging spray, spraying for 3 - 5 s every 1 h. Remove part of the shading net and restore the light intensity to 30%. Make the relative air humidity in the greenhouse reach 80 - 70%. Cultivate for 20 - 35 days. After the height of the seedlings generally reaches 2 cm, use foliar fertilizer (potassium dihydrogen phosphate) to promote the growth of the seedlings. Spray according to 0.1 - 1.0 g of foliar fertilizer / m 2Spray, so that the relative humidity of the air in the greenhouse reaches about 65%. After cultivating small rootstocks, take them out for transplantation, and the seedling survival rate of transplantation reaches 10%.

Claims

1. A method for large-scale seedling raising of Drynaria fortunei, and the specific steps are as follows: (1) Transplanting Drynaria fortunei in the greenhouse: Build a frame more than 10 cm above the ground in the greenhouse, then evenly spread camphor sawdust on it with a thickness of 8 - 20 cm as the greenhouse substrate, collect the rhizomes of Drynaria fortunei that have grown for many years and transplant them into the greenhouse substrate, use a shading net to reduce the light intensity to 30%, and continuously spray water to make the relative air humidity reach more than 75%; (2) Pretreatment before seedling raising: When the leaves of Drynaria fortunei to be cultivated start to change color, evenly sprinkle camphor sawdust on the greenhouse substrate with a thickness of 4 - 8 cm; (3) Collecting Drynaria fortunei spores: After the Drynaria fortunei spores mature, use tweezers or a small wooden stick to tap the Drynaria fortunei leaves to make the spores fall as evenly as possible on the camphor sawdust; (4) Spore germination: Use a shading net to reduce the light intensity to less than 15% again, start spraying germination nutrient solution by hanging spray during the day, spray for 3 - 5 s every 1 h, make the relative air humidity in the greenhouse reach more than 100%, and culture for 20 - 35 days; (5) Promoting rhizome formation: 10 days after the spores start to germinate, start spraying rhizome-promoting nutrient solution by hanging spray during the day, spray for 3 - 5 s every 1 h, remove part of the shading net, restore the light intensity to 30%, make the relative air humidity in the greenhouse reach 80 - 70%, and culture for 20 - 35 days; (6)Conventional management of seedlings: After the height of the seedlings generally reaches 2 cm, stop spraying the nutrient solution for promoting rhizome growth, and use foliar fertilizer spraying to promote the growth of seedlings. Spray according to 0.1 - 1.0 g of foliar fertilizer / m 2 Perform spraying. When the relative humidity of the air in the greenhouse reaches about 65%, cultivate until there are small rhizomes, and then take them out for transplanting.

2. According to the method of claim 1, the composition of the germination nutrient solution in step (4) is an aqueous solution of 0.4 - 1.2 g / L potassium nitrate, 0.1 - 0.4 g / L potassium dihydrogen phosphate, 50 - 60 mg / L inositol, 5 - 10 mg / L glycine, 0.5 - 1 mg / L nicotinic acid, 0.01 - 0.1 mg / L pyridoxine hydrochloride, 25 - 50 g / L thiophanate-methyl, and the pH value is about 5.

5.

3. According to the method of claim 1, the composition of the rhizome-promoting nutrient solution in step (5) is an aqueous solution of 0.06 - 0.15 g / L naphthylacetic acid, 0.4 - 1.2 g / L potassium nitrate, 0.1 - 0.4 g / L potassium dihydrogen phosphate, 50 - 60 mg / L inositol, 5 - 10 mg / L glycine, 0.5 - 1 mg / L nicotinic acid, 0.01 - 0.1 mg / L pyridoxine hydrochloride, 25 - 50 g / L thiophanate-methyl, and the pH value is about 5.5.

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