Method for rapidly breeding seedlings by utilizing edible tulip seeds

Abstract

The invention relates to a method for rapidly breeding seedlings by utilizing edible tulip seeds, and the method specifically comprises the steps of edible tulip seed pretreatment, germination accelerating treatment, separation, long leaf induction treatment, root induction treatment, edible tulip seedling management and the like. The seedling production cost and the seedling technical difficulty of the pseudobulbus cremastrae seu pleiones are greatly reduced; seedling raising is conducted through the germination accelerating device, 98% of the pseudobulbus cremastrae seu pleiones seeds are completely accelerated successfully, the seedling raising success rate of the pseudobulbus cremastrae seu pleiones seeds is greatly increased, the seedling rate of the pseudobulbus cremastrae seu pleiones seeds is further increased through continuous treatment of the leaf breeding and root breeding solution in the later period, and the purpose of large-scale seedling raising is achieved.

Description

Technical Field

[0001] The present invention specifically relates to a method for rapidly breeding seedlings by using Cremastra appendiculata seeds, belonging to the technical field of Chinese medicinal material cultivation. Background Art

[0002] Cremastra appendiculata is a perennial herb with clustered pseudobulbs. It grows in moist areas by ditches or under shaded forests on slopes below [altitude] meters, and is distributed in Tibet, Guizhou, Sichuan, northern Guangdong, Hunan, Hubei, Henan, Taiwan, Anhui, Jiangsu, Jiangxi, Zhejiang, southern Gansu, southern Shaanxi, and southern Shanxi. As the pseudobulb of Cremastra appendiculata, it tastes sweet and slightly pungent, with a small amount of toxicity, enters the liver and spleen meridians, and has the effects of detumescence, dissipating nodules, resolving phlegm, clearing heat, and detoxifying, and is widely used in anti-tumor drugs. Its growth form is an irregular flat spherical or conical shape, with a gradually protruding top, fibrous root scars at the base, about [length], and the diameter of the swollen part. The surface is yellowish-brown or brownish-black, with longitudinal wrinkles or grooves, and there are [number] slightly protruding joints in the middle, and filamentous fibers left after the scale leaves wither and rot on the joints. The texture is hard and difficult to break, and the cross-section is grayish-white or yellowish-white, slightly keratin-like. It has a faint smell, a light taste, and is viscous.

[0003] Cremastra appendiculata has high ornamental and medicinal values. Research on its active ingredients, pharmacological effects, and clinical applications shows that it has high biological activity, wide uses, and definite curative effects, and is worthy of further promotion and application, with high economic value. However, as a rare germplasm resource of medicinal plants in the Orchidaceae family, the self-pollination germination rate of its seeds is relatively low. Generally, artificial assisted pollination is required to obtain seeds, and the seeds are very small and difficult to develop into seedlings under natural conditions. In addition, Cremastra appendiculata mainly relies on pseudobulbs for reproduction in the natural state. One pseudobulb only grows one new pseudobulb in a year, and the reproduction is difficult. The traditional ramet cultivation has a long cycle, low efficiency, and a slow breeding process, and cannot be widely promoted.

[0004] To reduce the collection pressure on wild Orchidaceae plants and protect the biodiversity of Orchidaceae plants, it is urgent and necessary to research and develop plant tissue culture techniques with a short reproduction cycle and a high reproduction coefficient. In recent years, the research on the rapid propagation system for obtaining excellent Cremastra appendiculata seedlings by tissue culture technology has been booming. In the current research on Cremastra appendiculata, scholars mainly explore fields such as aseptic germination of seeds, bulb induction, callus cultivation, and obtaining cluster buds, and there is no special research on the morphological development and rapid propagation system of tissue bud blocks. Therefore, this project uses young tissue bud blocks as explants, and uses growth rate, weight gain rate, and growth form as indicators to explore influencing factors such as basic culture medium, content of macronutrients in the basic culture medium, single hormone, and hormone combination, and analyzes from the perspectives of morphological development and microscopic level, and systematically and comprehensively establishes a rapid propagation system for Cremastra appendiculata, which has high social and economic benefits.

[0005] The patent with the publication number CN106856725A discloses a method for germinating the seeds of Iphigenia indica. Select the fruit pods of Iphigenia indica that are mature in the current year, rub them to obtain the seeds, and sow the seeds on the surface of a sowing substrate which has passed through a 40-mesh sieve and is a uniform mixture of mountain base soil and peat soil in a ratio of 1:1. Germinate the seeds under constant temperature conditions of 20°C with light exposure, and keep the surface of the germination substrate moist. After 15 - 18 days, the seeds take root and germinate, and then cotyledons grow out after 3 - 5 days. The comprehensive seedling formation rate is about 80% - 85%. Due to the lack of endosperm in the seeds of Iphigenia indica, the seedling formation rate is extremely low, and it is impossible to produce seedlings on a large scale.

[0006] The patent with the publication number CN109076958A discloses a sterile propagation technology of Iphigenia indica in vitro. Select mature unopened capsules, disinfect the surface with 0.1% mercuric chloride successively, wash with sterile water, dry with sterile filter paper to remove excess water and then air-dry. Then, cut the capsules under sterile conditions and sow the seeds in a glassware filled with culture medium; add 1 mL of 0.1% agar to each bottle of culture medium, shake the bottle body to mix the seeds evenly and spread them flat on the culture medium, and then wrap the whole glassware tightly with newspaper until it is light-proof. Let it stand for 72 h under the condition of 26°C in the room; take out the seeds after standing, mix them evenly with disinfected fine sand in a ratio of 1:5, and then evenly sprinkle them into a test tube filled with moist seedling-raising substrate. The fine sand can be sterilized by high-temperature sterilization; place the test tube in a 25°C culture room for aseptic culture. During the stage of inducing seed germination, dark culture is adopted. After about 15 days of culture, the seeds of Iphigenia indica begin to germinate. At this time, the embryo breaks through the seed coat, the volume increases significantly and a white proembryonic bulb is formed; transplant the germinated Iphigenia indica to a nursery prepared in advance for acclimatization and strengthening of seedlings two months after germination; the nursery substrate can be humus soil; immediately spray a carbendazim agent once after transplantation; then spray the full-dose MS culture solution once every 3 days to root and strengthen the seedlings; control the temperature in the greenhouse above 20°C. Although the seedling formation rate is high by using tissue culture technology, there are technical obstacles and the input cost is relatively high, and the production cost of seedlings is high.

[0007] The patent with the publication number CN109496749A discloses a cultivation method of Cremastra appendiculata. First, take out the Cremastra appendiculata seeds stored in sand in the previous year, disinfect and sterilize them, and then place them in a warm and humid dark room for 4 - 5 hours. Then, after the seeds show white tips, put the seeds in a flexible mesh bag and keep moisturizing and shaking them for 10 - 12 hours to promote the splitting of the embryo. Finally, soak the seeds in water for 1.5 - 2 days, then take them out, soak them in carbendazim for 20 - 30 minutes for sterilization, and then sow them in the seedling raising room for constant temperature and humidity seedling raising. When the seedling emergence rate of the seeds reaches 70%, cover the first layer of pine needles on the seedlings, with the covering thickness not exceeding 1 cm and the covering density such that the visible seedling rate reaches 30 - 40%. When the seedling emergence rate on the first layer of pine needles reaches 50%, cover the second layer of pine needles, with the covering thickness not exceeding 0.5 cm and the covering density such that the visible seedling rate reaches 60 - 70%. Select the Cremastra appendiculata seedlings that emerge from the second layer of pine needles for transplantation 10 - 15 days after the second layer of pine needles starts to show seedlings, and then carry out subsequent cultivation with conventional field management. Since the Cremastra appendiculata seeds lack endosperm, there are also problems such as extremely low seedling formation rate.

[0008] The patent with the publication number CN110463608A discloses a new method for rapid artificial propagation of Cremastra appendiculata. Select Cremastra appendiculata plants with excellent phenotypic traits and take their pseudobulbs as explants. First, wash the surface soil with tap water, then soak it in a washing powder solution, shake and stir it, and then rinse it with running water and place it on the workbench. Then soak it in a vitamin C solution, then treat it with an ethanol solution, then disinfect it with a mercury aqueous solution, and finally rinse it to obtain sterilized pseudobulbs. Cut the stem segments of the sterilized pseudobulbs according to the bud points with a scalpel and put them into medium A for cultivation. Transfer the young buds grown from the bud points into medium A in step 3 for proliferation culture. At the same time, it is found that callus will be produced at the base of the explants in step 3. Cut off the seedlings with callus and continue to transfer them into medium B for cultivation. Cultivate under the conditions of controlling light intensity, temperature, and light time, and synchronously induce callus, initiate and proliferate cluster buds. Then observe that the callus begins to proliferate. When purple bud points appear on the surface of the callus, the callus rapidly differentiates into adventitious buds, the adventitious buds begin to differentiate into seedlings, and adventitious roots occur. Then transfer the callus to the same medium B for continuous proliferation and differentiation, and transfer the obtained seedlings to the same medium B for continuous cultivation. Small pseudobulbs the size of rice grains appear at the base of the test-tube seedlings. Then inoculate the single seedlings with small pseudobulbs the size of rice grains after subculture into medium C for cultivation. Cultivate under the conditions of controlling light intensity, temperature, and light time until complete roots are formed. Acclimatize the seedlings at room temperature. Take out the seedlings from medium C, wash the remaining medium C clean, disinfect them in a carbendazim solution, and then transplant them into the sterilized substrate for heat preservation and moisture preservation cultivation to obtain transplanted seedlings. This technology also has problems such as tissue culture costs, and for the seedlings formed by callus differentiation, there are problems such as unstable gene inheritance.

[0009] Liao Tingting et al. in "Establishment of a Rapid Propagation System for Cremastra appendiculata" used the tissue bud explants of Cremastra appendiculata as test materials to explore the most suitable cultivation plan for Cremastra appendiculata and establish a rapid propagation system for Cremastra appendiculata. Feng Fosheng et al. in "Rapid Propagation of Cremastra appendiculata" analyzed the composition changes after one or two times of steaming processing and then pulverization. Using the corms and mature seeds of Cremastra appendiculata as materials, different culture media and culture conditions were adopted to study the most suitable breeding conditions and the technical system for large-scale production of seedlings.

[0010] From the existing literature retrieval, it is found that in the research on the rapid propagation of Cremastra appendiculata, the technology of directly sowing Cremastra appendiculata seeds for high-efficiency seedling production has not been reported. There are two main difficulties: one is that when raising seedlings with Cremastra appendiculata seeds, it is difficult to form seedlings due to the lack of endosperm; the other is that the small seedlings grown from Cremastra appendiculata seeds grow relatively slowly and are not suitable for the current needs of medicinal farmers. Summary of the Invention

[0011] In view of the above problems, the present invention provides a method for rapidly propagating seedlings using Cremastra appendiculata seeds, which can effectively solve the problem of low seedling formation rate during the breeding of Cremastra appendiculata seeds, and can also scale down the production cost of seedlings, meeting the requirements of industrial production, with advantages such as uniform seedling size, strong operability, and high seedling formation rate.

[0012] The object of the present invention is to provide a method for rapidly propagating seedlings using Cremastra appendiculata seeds, and the specific steps are as follows:

[0013] (1) Pretreatment of Cremastra appendiculata seeds: Put the Cremastra appendiculata seeds (from January to March every year) into the pretreatment solution and soak for 3 - 6 h, with 3 - 5 oscillations at intervals during the soaking process, and the temperature is 15 - 20 °C;

[0014] (2) Germination promotion treatment: Place the pretreated Cremastra appendiculata seeds in a germination promotion machine, soak them 20 - 40 times a day with the germination promotion solution, the soaking time is 3 - 5 min, the temperature is maintained at 15 - 20 °C, and the light intensity is 500 - 700 LX;

[0015] (3) Separation: Separate the Cremastra appendiculata seeds (not germinated) floating on the water surface from the germinated Cremastra appendiculata seeds (existing in the water body or sinking to the bottom of the water body), and place the non-germinated Cremastra appendiculata seeds back into the germination promotion device to continue germination promotion;

[0016] (4) Long leaf induction treatment: Sprinkle the germinated Cremastra appendiculata seeds directly on the sterilized seedling raising substrate with a watering can, maintain the relative humidity at 70 - 95%, control the temperature at 15 - 20 °C, the light intensity at 200 - 400 LX, spray the leaf-growing solution 0.5 - 2 kg per square meter each time, spray 2 - 4 times a day, and maintain for 20 - 35 days;

[0017] (5) Inducing root treatment: The seeds of Cremastra appendiculata that have started to grow leaves are treated with a root-growing solution three times. The interval between the three treatments is 3 - 5 days. Each time, 1 - 4 kg / m² of the root-growing solution is sprayed. The cultivation temperature is 15 - 20°C, the daily light exposure is 5 - 8 h, the light intensity is 200 - 400 LX, the relative humidity is controlled at 50 - 65%, and the cultivation lasts for 35 - 45 days.

[0018] (6) Management of Cremastra appendiculata seedlings: After the Cremastra appendiculata grows roots, at 15 - 20°C, with a daily light exposure of 5 - 8 h, a light intensity of 200 - 400 LX, and the relative humidity controlled at 50 - 70%, 0.5 - 1 kg / m² of a nutrient solution is sprayed once, and then sprayed once every 1 - 2 days. In October - December, the one-year-old Cremastra appendiculata seedlings that have withered are taken out and transplanted to the field or other seedbeds for planting.

[0019] In a specific embodiment, the pretreatment solution in step (1) refers to an aqueous solution containing 80 - 100 mg / L of 6-benzylaminopurine and 30 - 50 g / L of carbendazim.

[0020] In a specific embodiment, the germination-inducing solution in step (2) contains an aqueous solution of 50 - 80 mg / L of 6-benzylaminopurine, 20 - 40 mg / L of gibberellin, and 30 - 40 g / L of carbendazim.

[0021] In a specific embodiment, the seedling-raising substrate in step (4) refers to a substrate with 3 - 5 cm thick fine sand at the bottom layer, covered with 3 - 5 cm thick coconut coir on the upper layer, and then sterilized with a 6 - 10% carbendazim solution.

[0022] In a specific embodiment, the leaf-growing solution in step (4) refers to an aqueous solution containing 50 - 70 mg / L of 6-benzylaminopurine, 1 - 1.5 g / L of ammonium nitrate, 1.5 - 1.8 g / L of potassium nitrate, 200 - 350 mg / L of calcium chloride, 150 - 250 mg / L of magnesium sulfate, 100 - 160 mg / L of potassium dihydrogen phosphate, 80 - 100 mg / L of inositol, 0.3 - 0.5 mg / L of nicotinic acid, 0.3 - 0.5 mg / L of vitamin B6, 0.06 - 0.08 mg / L of vitamin B1, 1 - 1.5 mg / L of glycine, and 30 - 40 g / L of carbendazim, with a pH of 5.5 - 6.

[0023] In a specific embodiment, the root-growing solution in step (5) refers to an aqueous solution containing 80 - 100 mg / L of naphthylacetic acid, 1 - 1.5 g / L of ammonium nitrate, 1.5 - 1.8 g / L of potassium nitrate, 200 - 350 mg / L of calcium chloride, 150 - 250 mg / L of magnesium sulfate, 100 - 160 mg / L of potassium dihydrogen phosphate, 80 - 100 mg / L of inositol, 0.3 - 0.5 mg / L of nicotinic acid, 0.3 - 0.5 mg / L of vitamin B6, 0.06 - 0.08 mg / L of vitamin B1, 1 - 1.5 mg / L of glycine, and 25 - 35 g / L of carbendazim, with a pH of 5.5 - 6.

[0024] The preparation method of the above carbendazim solution is to weigh 7 g of carbendazim powder purchased from the market and add 93 g of water for mixing to obtain a 7% carbendazim solution.

[0025] In a specific embodiment, the nutrient solution in step (6) is an aqueous solution containing 0.3 - 1 g / L of ammonium nitrate, 1 - 1.5 g / L of potassium nitrate, 0.1 - 0.8 g / L of potassium dihydrogen phosphate, 40 - 70 mg / L of inositol, 0.3 - 0.5 mg / L of nicotinic acid, 0.3 - 0.5 mg / L of vitamin B6, 0.06 - 0.08 mg / L of vitamin B1, and 0.8 - 1.2 mg / L of glycine, with a pH value of 5.5 - 6.0.

[0026] All of the above solutions are prepared using mountain spring water.

[0027] The second object of the present invention is to provide the above-mentioned germinating machine, which consists of four parts: a separation structure, a constant temperature water circulation structure, a lighting structure, and a box body. The separation structure includes a perforated plate and an intermediate valve. The constant temperature water circulation structure includes a circulation water pump, a heating wire, a water injection pipe, a float valve, and a float. The water at the bottom of the box body is pumped out through the float valve and the float by the circulation water pump, and then introduced onto the perforated plate through the water injection pipe. The valve is opened or closed to control the water flow rate, and the temperature of the water at the bottom of the box body is kept constant by the heating wire. The lighting structure includes a lid, a fixing frame, and a lamp tube. The lamp tube is fixed on the lid through the fixing frame.

[0028] Preferably, the pore size of the perforated plate is greater than 1 mm and less than 5 cm.

[0029] Preferably, when the perforated plate is in use, two to four layers of filter paper are placed on the perforated plate, and the filter paper is fixed on the perforated plate with a layer of gauze.

[0030] Technical effects

[0031] 1. For the first time, direct seeding of Cremastra appendiculata seeds is used for seedling raising, achieving greenhouse seedling establishment under the condition of seeds without endosperm, greatly reducing the production cost and technical difficulty of Cremastra appendiculata seedling raising.

[0032] 2. For the first time, a self-made germinating machine was used for heat and moisture preservation during seedling raising, achieving a 98% successful germination rate of Cremastra appendiculata seeds, greatly improving the success rate of Cremastra appendiculata seedling raising. Subsequently, through continuous treatment with leaf and root growth solutions, the seedling survival rate of Cremastra appendiculata seeds was further increased, achieving the goal of large-scale seedling raising. Description of the Drawings

[0033] Figure 1 : Schematic diagram of the germinating device, where 1. Lid; 2. Fixed frame; 3. Lamp tube; 4. Water injection pipe; 5. Floating ball; 6. Circulating water pump; 7. Float valve; 8. Perforated plate; 9. Metal plate; 10. Valve; 11. Box body; 12. Heating wire;

[0034] Figure 2 : Newly germinated Cremastra appendiculata seeds;

[0035] Figure 3 : One-year-old Cremastra appendiculata seedling. Detailed Implementation Manner

[0036] Next, the present invention will be further illustrated with examples, but it is not limited to any one or similar examples of these examples.

[0037] Example 1: Implementation Description of the Germinating Device

[0038] As Figure 1 shown, the implementation process of this germinating device is as follows: The lamp tube 3 is fixed on the lid 1 through the fixed frame 2 (providing light for Cremastra appendiculata seeds). Inside the box body 11, two to four layers of filter paper are placed on the perforated plate 8 (with a pore diameter greater than 1 mm and less than 5 cm), and the filter paper is fixed on the perforated plate with a layer of gauze. The Cremastra appendiculata seeds are placed on it. The circulating water pump 6 is used to pump the water at the bottom of the box body 11 through the float valve 7 and the floating ball 5, and then introduce it into the filter paper through the water injection pipe 4. At the same time, the valve 12 is closed, so that the Cremastra appendiculata seeds can be immersed in water. Then, by opening the valve 12, the soaking time of the Cremastra appendiculata seeds can be controlled, and the temperature of the water at the bottom of the box body 11 is controlled to be constant during germination by the heating wire 12; the box body 11 forms upper and lower spaces with the valve 10 through the metal plate 9, and the upper and lower spaces are connected through the valve 10.

[0039] Example 2: Seedling Raising of Cremastra appendiculata Seeds

[0040] On November 18th, the Cremastra appendiculata seeds were taken and soaked in the pretreatment solution for 3 - 6 h, with intermittent oscillation 3 - 5 times during the soaking process, and the temperature was 15 - 20 °C; the pretreated Cremastra appendiculata seeds were placed in the germinating device ( Figure 1 ) and soaked 30 times a day with the germinating solution, the soaking time was 3 - 5 min, the temperature was maintained at 15 - 20 °C, and the light intensity was 500 - 700 LX; the ungerminated Cremastra appendiculata seeds floating on the water surface were separated from the germinated Cremastra appendiculata seeds (Figure 2 ) Separate them, place the ungerminated Cremastra appendiculata seeds back into the germination-promoting device to continue germination promotion; sprinkle the germinated Cremastra appendiculata seeds directly onto the sterilized seedling-raising substrate with a watering can, maintain the relative humidity at 70 - 95%, control the temperature at 15 - 20°C, the light intensity at 300 LX, spray the leaf-growing solution 0.5 - 2 kg per square meter each time, spray 2 - 4 times a day, and maintain for 25 days; treat the Cremastra appendiculata seedling that has started to grow leaves with the root-growing solution three times in total, with an interval of 3 - 5 days between the three treatments, spray the root-growing solution 1 - 4 kg per square meter each time, the cultivation temperature is 15 - 20°C, with 5 - 8 hours of light per day, the light intensity is 300 LX, and the relative humidity is controlled at 50 - 65%, cultivate for 35 - 45 days; after the Cremastra appendiculata grows roots, at 15 - 20°C, with 5 - 8 hours of light per day, the light intensity is 200 - 400 LX, the relative humidity is controlled at 50 - 70%, spray the nutrient solution 0.5 - 1 kg per square meter once, and spray once every 2 days (see Figure 3 ), in November, take out the one-year-old Cremastra appendiculata seedling with withered leaves, conduct statistics, the seedling survival rate only reaches 94.2%, about 96% of its pseudobulb diameter reaches more than 1.5 cm, and then transplant it to the field or other seedling beds for planting, and the transplant survival rate reaches more than 99%.

[0041] Control Example 1

[0042] Soak the Cremastra appendiculata seeds taken on November 18th in the pretreatment solution for 3 - 6 hours, shake them 3 - 5 times at intervals during the soaking process, and the temperature is 15 - 20°C; sprinkle the pretreated Cremastra appendiculata seeds directly onto the sterilized seedling-raising substrate with a watering can, maintain the relative humidity at 70 - 95%, control the temperature at 15 - 20°C, the light intensity at 300 LX, spray the leaf-growing solution 0.5 - 2 kg per square meter each time, spray 2 - 4 times a day, and maintain for 25 days; treat the Cremastra appendiculata seedling that has started to grow leaves with the root-growing solution three times in total, with an interval of 3 - 5 days between the three treatments, spray the root-growing solution 1 - 4 kg per square meter each time, the cultivation temperature is 15 - 20°C, with 5 - 8 hours of light per day, the light intensity is 300 LX, and the relative humidity is controlled at 50 - 65%, cultivate for 35 - 45 days; after the Cremastra appendiculata grows roots, at 15 - 20°C, with 5 - 8 hours of light per day, the light intensity is 200 - 400 LX, the relative humidity is controlled at 50 - 70%, spray the nutrient solution 0.5 - 1 kg per square meter once, and spray once every 2 days. In November, take out the one-year-old Cremastra appendiculata seedling with withered leaves, conduct statistics, the seedling survival rate only reaches 21.6%, about 18.5% of its pseudobulb diameter reaches more than 1.5 cm, and then transplant it to the field or other seedling beds for planting, and the transplant survival rate reaches more than 99%.

[0043] Control Example 2

[0044] On November 18th, pick the seeds of Cremastra appendiculata and soak them in the pretreatment solution for 3 - 6 hours. During the soaking process, shake them 3 - 5 times at intervals, and keep the temperature at 15 - 20°C. Place the pretreated seeds of Cremastra appendiculata in the germination device, soak them 30 times a day with the germination solution, with the soaking time being 3 - 5 minutes, the temperature maintained at 15 - 20°C, and the light intensity at 500 - 700 LX. When most of the seeds of Cremastra appendiculata have germinated, directly spray them onto the sterilized seedling substrate with a watering can, maintain the relative humidity at 70 - 95%, control the temperature at 15 - 20°C, and the light intensity at 300 LX. Spray the leaf-growing solution at 0.5 - 2 kg per square meter each time, spray 2 - 4 times a day, and maintain for 25 days. Treat the seedling of Cremastra appendiculata that has started to grow leaves with the root-growing solution three times in total. The interval between the three treatments is 3 - 5 days. Spray the root-growing solution at 1 - 4 kg per square meter each time, with the cultivation temperature at 15 - 20°C, 5 - 8 hours of light per day, the light intensity at 300 LX, and the relative humidity controlled at 50 - 65%. Cultivate for 35 - 45 days. After the roots of Cremastra appendiculata grow, at 15 - 20°C, 5 - 8 hours of light per day, the light intensity at 200 - 400 LX, and the relative humidity controlled at 50 - 70%, spray the nutrient solution at 0.5 - 1 kg per square meter once, and spray again at an interval of 2 days. In November, take out the one-year-old seedling of Cremastra appendiculata that has withered and conduct statistics. The seedling survival rate only reaches 87.8%. About 95.4% of them have a pseudobulb diameter of more than 1.5 cm. Then transplant them to the field or other seedbeds for planting, and the transplanting survival rate reaches more than 99%.

Claims

1. A method for rapidly propagating seedlings using Cremastra appendiculata seeds, the specific steps are as follows: (1) Pretreatment of Cremastra appendiculata seeds: Put the Cremastra appendiculata seeds into a pretreatment solution and soak for 3 - 6 h, with intermittent oscillation 3 - 5 times during the soaking process, and the temperature is 15 - 20 °C; (2) Germination promotion treatment: Place the pretreated Cremastra appendiculata seeds in a germination promotion device, soak with a germination promotion solution 20 - 40 times a day, the soaking time is 3 - 5 min, the temperature is maintained at 15 - 20 °C, and the light intensity is 500 - 700 LX; (3) Separation: Separate the Cremastra appendiculata seeds floating on the water surface from the germinated Cremastra appendiculata seeds, and place the ungerminated Cremastra appendiculata seeds back in the germination promotion device to continue germination promotion; (4) Leaf growth induction treatment: Sprinkle the germinated Cremastra appendiculata seeds directly on the sterilized seedling-raising substrate with a watering can, maintain the relative humidity at 70 - 95%, control the temperature at 15 - 20 °C, the light intensity at 200 - 400 LX, spray the leaf-growing solution 0.5 - 2 kg per square meter each time, spray 2 - 4 times a day, and maintain for 20 - 35 days; (5) Root induction treatment: Treat the Cremastra appendiculata seedling with the root-growing solution three times in total, with an interval of 3 - 5 days between the three treatments, spray the root-growing solution 1 - 4 kg per square meter each time, the cultivation temperature is 15 - 20 °C, the daily light is 5 - 8 h, the light intensity is 200 - 400 LX, and the relative humidity is controlled at 50 - 65%, and cultivate for 35 - 45 days; (6) Management of Cremastra appendiculata seedlings: After the Cremastra appendiculata grows roots, at 15 - 20 °C, the daily light is 5 - 8 h, the light intensity is 200 - 400 LX, the relative humidity is controlled at 50 - 70%, spray the nutrient solution 0.5 - 1 kg per square meter once, spray once every 1 - 2 days, and take out the one-year-old Cremastra appendiculata seedling with withered seedlings in October - December for transplanting to the field or other seedling beds for planting.

2. According to the method of claim 1, the pretreatment solution in step (1) refers to an aqueous solution containing 80 - 100 mg / L 6-benzylaminopurine and 30 - 50 g / L carbendazim.

3. According to the method of claim 1, the germination promotion solution in step (2) contains an aqueous solution of 50 - 80 mg / L 6-benzylaminopurine, 20 - 40 mg / L gibberellin, and 30 - 40 g / L carbendazim.

4. According to the method of claim 1, the seedling-raising substrate in step (4) refers to being covered with 3 - 5 cm thick fine sand at the lower layer and 3 - 5 cm coconut bran at the upper layer, and then sterilized with a 6 - 10% carbendazim solution.

5. According to the method of claim 1, the leaf-growing solution in step (4) refers to an aqueous solution containing 50 - 70 mg / L 6-benzylaminopurine, 1 - 1.5 g / L ammonium nitrate, 1.5 - 1.8 g / L potassium nitrate, 200 - 350 mg / L calcium chloride, 150 - 250 mg / L magnesium sulfate, 100 - 160 mg / L potassium dihydrogen phosphate, 80 - 100 mg / L inositol, 0.3 - 0.5 mg / L nicotinic acid, 0.3 - 0.5 mg / L vitamin B6, 0.06 - 0.08 mg / L vitamin B1, 1 - 1.5 mg / L glycine, and 30 - 40 g / L carbendazim, with a pH of 5.5 - 6.

6. According to the method of claim 1, the root-growing solution in step (5) refers to an aqueous solution containing 80 - 100 mg / L naphthylacetic acid, 1 - 1.5 g / L ammonium nitrate, 1.5 - 1.8 g / L potassium nitrate, 200 - 350 mg / L calcium chloride, 150 - 250 mg / L magnesium sulfate, 100 - 160 mg / L potassium dihydrogen phosphate, 80 - 100 mg / L inositol, 0.3 - 0.5 mg / L nicotinic acid, 0.3 - 0.5 mg / L vitamin B6, 0.06 - 0.08 mg / L vitamin B1, 1 - 1.5 mg / L glycine, and 25 - 35 g / L carbendazim, with a pH of 5.5 - 6.

7. According to the method of claim 1, the nutrient solution in step (6) is an aqueous solution containing 0.3 - 1 g / L ammonium nitrate, 1 - 1.5 g / L potassium nitrate, 0.1 - 0.8 g / L potassium dihydrogen phosphate, 40 - 70 mg / L inositol, 0.3 - 0.5 mg / L nicotinic acid, 0.3 - 0.5 mg / L vitamin B6, 0.06 - 0.08 mg / L vitamin B1, and 0.8 - 1.2 mg / L glycine, with a pH value of 5.5 - 6.0.

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