A method for rapid propagation and large-scale cultivation of Anubias

Abstract

The present invention provides a method for rapid propagation and large-scale cultivation of Anubias barteri var. nana, comprising the following steps: selecting young flower stalks of Anubias barteri var. nana that have not yet extended out of the leaf sheaths, removing the parts at both ends that have come into contact with the disinfectant after disinfection; cutting into thin slices and inoculating them on an adventitious bud medium for dark culture to induce adventitious buds; cutting the adventitious buds into small clusters and inoculating them on a proliferation medium for proliferation culture; cutting the proliferated buds with leaves exhibited into single plants, inoculating them on a rooting medium, and obtaining tissue culture seedlings through rooting culture; washing the roots of the tissue culture seedlings and wrapping them with rock wool, fixing the rock wool, adding water to submerge the leaves of the tissue culture seedlings, and reducing the water level after new leaves grow; water and fertilizer management. The present invention uses young flower stalks that have not yet extended out of the leaf sheaths as explants, with a high success rate of disinfection, fast recovery of growth, short subculture cycle, high proliferation coefficient, and short rooting time; uses rock wool as the cultivation substrate, and sprays water for hydroponics regularly, with the seedlings growing neatly and quickly, and large-scale production of Anubias barteri var. nana can be achieved.

Description

Technical Field

[0001] The present invention belongs to the field of seedling production, and particularly relates to a method for rapid propagation and large-scale cultivation of Anubias barteri Background Art

[0002] Anubias barteri is a general term for plants of the genus Anubias in the family Araceae. It is native to the tropical regions of West Africa and has strong plasticity in landscaping. It is one of the most widely used ornamental aquatic plants in aquariums. Its leaf surface is smooth, leaf color is dark green, leaf veins are clear, the base is heart-shaped, leathery, the leaves are like those of Ficus, and the roots are like the aerial roots of Ficus. Therefore, it is named 'Anubias barteri'. Planting in patches can give people a feeling of cool shade, and it is an indispensable aquatic plant for aquarium landscaping.

[0003] Anubias barteri is usually propagated by division. The propagation coefficient is low, the growth is slow, the cycle is long, and it is easy to carry diseases, which greatly limits the large-scale and commercial production of Anubias barteri. Through tissue culture technology, a large number of high-quality seedlings can be produced in a short time to meet the market demand.

[0004] Anubias barteri has been growing in a high-humidity environment for a long time, and its endophytic bacteria are serious, which brings difficulties to the rapid propagation of Anubias barteri by tissue culture. Domestic reports on the rapid propagation of Anubias barteri by tissue culture all use shoot tips or axillary buds as explants. Not only the disinfection time is long, which is easy to cause browning and death of the explants, but also the disinfection success rate is low. In addition, after multiple generations of culture, endophytic bacteria are likely to be produced, affecting the subsequent proliferation and seedling quality.

[0005] Anubias barteri tissue culture seedlings grow in a closed tissue culture bottle and are adapted to the high-humidity growth environment. They can be directly used for aquarium landscaping. However, this method is only applicable to small varieties of Anubias barteri. For medium and large varieties of Anubias barteri, the growth rate is slow, the cycle is long, and the landscaping effect is poor; in production, sand or aquatic plant mud is used as the substrate for hydroponic cultivation in greenhouses. However, this method not only has slow growth, high incidence of diseases, inconvenient operation, but also requires secondary planting before sales. Neither of the above two methods can achieve large-scale production to meet the market demand. Summary of the Invention

[0006] The purpose of the present invention is to overcome the deficiencies existing in the prior art and achieve large-scale production. The present invention provides a method for rapid propagation and cultivation of Anubias barteri.

[0007] A method for rapid propagation and cultivation of Anubias barteri, comprising the following steps: selecting young flower stalks of Anubias barteri that have not yet emerged from the leaf sheaths, disinfecting and removing the parts at both ends that have come into contact with the disinfectant; cutting into thin slices and inoculating them onto an adventitious bud medium LS + 3 - 5 mg / L 6-BA + 0.1 - 0.2 mg / L NAA + 2 - 4 mg / L inositol + 29 - 31 g / L sucrose + 5 - 6 g / L carrageenan, and performing dark culture for adventitious bud induction; cutting the adventitious buds into small clusters and inoculating them onto a proliferation medium LS + 2 - 4 mg / L 6-BA + 0.1 - 0.2 mg / L NAA + 29 - 31 g / L sucrose + 5 - 6 g / L carrageenan for proliferation culture; cutting the proliferated buds with leaves exhibited into single plants, inoculating them onto a rooting medium 1 / 2LS + 0.1 - 0.2 mg / L NAA + 29 - 31 g / L sucrose + 5 - 6 g / L carrageenan + 0.5 - 1 g / L activated carbon, and performing rooting culture to obtain tissue culture seedlings; washing the roots of the tissue culture seedlings and wrapping them with rock wool, fixing the rock wool, adding water to submerge the leaves of the tissue culture seedlings, and reducing the water level after new leaves grow; water and fertilizer management.

[0008] Preferably, the adventitious bud medium is LS + 4 mg / L 6-BA + 0.1 mg / L NAA + 3 mg / L inositol + 30 g / L sucrose + 5.5 g / L carrageenan.

[0009] Preferably, the proliferation medium is LS + 3 mg / L 6-BA + 0.1 mg / L NAA + 30 g / L sucrose + 5.5 g / L carrageenan.

[0010] Preferably, the rooting medium is 1 / 2LS + 0.1 mg / L NAA + 30 g / L sucrose + 5.5 g / L carrageenan + 0.5 g / L activated carbon.

[0011] Preferably, the components of the LS medium are as follows: 1650 mg / L NH4NO3, 1500 mg / L KNO3, 480 mg / L CaCl2·2H2O, 370 mg / L MgSO4·7H2O, 227 mg / L KH2PO4, 22.3 mg / L MnSO4·4H2O, 8.6 mg / L ZnSO4·7H2O, 6.2 mg / L H3BO3, 0.83 mg / L KI, 0.25 mg / L Na2MoO4·2H2O, 0.25 mg / L CuSO4·5H2O, 0.025 mg / L CoCl2, 37.13 mg / L FeSO4·7H2O, 49.67 mg / L Na2·EDTA, 100 mg / L inositol, 2.0 mg / L glycine, 0.1 mg / L thiamine hydrochloride, 0.5 mg / L nicotinic acid, 0.5 mg / L pyridoxine hydrochloride, pH 5.8, and the solvent is water.

[0012] Preferably, the thickness of the thin slices is 1 - 2 mm.

[0013] Preferably, the disinfection is as follows: After washing the impurities on the surface of the explant with tap water, place it in a laminar flow hood. First, disinfect it with 75% alcohol for 30 s to 1 min, wash it 3 times with sterile water, then disinfect it with 0.1% mercuric chloride for 8 min, shake it continuously during this period, and finally wash it 5 times with sterile water.

[0014] Preferably, 2 - 4 g of slow-release fertilizer (Osmocote) is added when wrapping with rock wool; the fixed rock wool is rolled into a cylindrical shape, the diameter of the rolled rock wool cylinder is the same as the diameter of the planting basket, placed in the planting basket, and the lowermost part of the rock wool closely adheres to the bottom of the planting basket.

[0015] Preferably, the water and fertilizer management is carried out by an automatic spraying facility. In summer and autumn, water is sprayed once every 1 - 1.5 h from 8:00 to 18:00, and the spraying time is 1 - 3 min until the entire leaf surface is wet. In winter and spring, water is sprayed once every 1.5 - 2 h from 9:00 to 17:00, and the spraying time is 1 - 3 min until the entire leaf surface is wet; after the tissue-cultured seedlings grow the first new leaf, start spraying with 1000-fold liquid compound fertilizer and 20000-fold liquid urea, and the spraying time for each time is 2 - 5 min, once every half month; the compound fertilizer has an elemental mass ratio of N:P:K = 15:15:15.

[0016] Preferably, the Anubias is Anubias barteri, Anubias acuminata, Anubias hastifolia, Anubias lanceolata, Ficus elastica.

[0017] Compared with the prior art, the present invention has the following advantages:

[0018] (1) The present invention uses the young pedicels that have not yet emerged from the leaf sheaths as explants, greatly shortening the disinfection time of mercuric chloride, minimizing the harm of heavy metals to the explants to the greatest extent, having a high disinfection success rate, and a fast growth recovery.

[0019] (2) The present invention adds inositol during the bud induction period, which can induce adventitious buds faster and better.

[0020] (3) The present invention uses LS as the basic medium during the proliferation culture and rooting culture periods, shortening the subculture cycle and the rooting culture time, having a higher proliferation coefficient, and the grown seedlings are also stronger.

[0021] (4) The method of the present invention uses rock wool as the cultivation substrate during cultivation and adopts the method of timed spray hydroponic cultivation. Rock wool has undergone high-temperature calcination during production, will not carry pathogens and microorganisms, and has strong water absorption and water retention capabilities, which can create the best growth environment for plant roots; timed spray cultivation can not only meet the requirements of Anubias for a high-humidity growth environment but also facilitate the application of foliar fertilizers. The method of the present invention saves time and effort, is convenient for management, the growth of the seedlings is neat and fast, and the large-scale production of Anubias can be realized. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 It is a young pedicel.

[0023] Figure 2 It is an induced adventitious bud cluster.

[0024] Figure 3 It is a proliferated in-vitro seedling.

[0025] Figure 4 It is a rooted in-vitro seedling. Specific implementation mode

[0026] The following examples are further illustrations of the present invention, rather than limitations thereof.

[0027] The water ferns selected in the examples and comparative examples 1-7 of the present invention all include Anubias barteri, Anubias lanceolata, Anubias hastifolia, Anubias hastifolia var. lanceolata, Anubias nana.

[0028] Example:

[0029] (1) Explant preparation: Select healthy water fern plants without pests and diseases and whose inflorescences have not yet emerged from the leaf sheaths, peel off the external leaves, and take out the young pedicels as explants ( Figure 1 ).

[0030] (2) Explant disinfection: After washing off the impurities on the surface of the explants with tap water, place them in a laminar flow hood for disinfection. First, disinfect with 75% alcohol for 30 s - 1 min, wash 3 times with sterile water, then disinfect with 0.1% mercuric chloride for 8 min, shake constantly during this period, and finally wash 5 times with sterile water.

[0031] (3) Induction of adventitious buds: Cut off the parts of the disinfected explants that have contacted the disinfectant at both ends, and then cut them into slices with a thickness of about 1 mm, and inoculate them into a medium of LS + 4 mg / L 6 - BA + 0.1 mg / L NAA + 3 mg / L inositol + 30 g / L sucrose + 5.5 g / L carrageenan. After inoculation, place them in an incubator for dark culture, and the culture temperature is 25 ± 2 °C.

[0032] (4) Proliferation culture: Cut the induced adventitious bud clusters ( Figure 2 ) into small clusters of 1 cm 2 , and inoculate them into a medium of LS + 3 mg / L 6 - BA + 0.1 mg / L NAA + 30 g / L sucrose + 5.5 g / L carrageenan. After inoculation, place them in an incubator for culture, and the culture temperature is 25 ± 2 °C. Perform dark culture for the first 3 days, and then irradiate with light for 6 h every day, and the light intensity is 1000 lux. The proliferated in-vitro seedlings after proliferation culture are as shown in Figure 3 .

[0033] (5) Rooting culture: Cut the proliferated buds with good growth and 2 leaves showing into single plants and inoculate them into the rooting medium. The rooting medium is: 1 / 2LS + 0.1 mg / L NAA + 30 g / L sucrose + 5.5 g / L carrageenan + 0.5 g / L activated carbon. After inoculation, place them in the culture room for cultivation. The cultivation temperature is 25 ± 2 °C, with dark cultivation for the first 3 days, and then 10 h of light per day with a light intensity of 2000 lux. The rooted bottle seedlings after rooting culture are as Figure 4 shown.

[0034] The LS medium contains the following components: 1650 mg / L NH4NO3, 1500 mg / L KNO3, 480 mg / L CaCl2·2H2O, 370 mg / L MgSO4·7H2O, 227 mg / L KH2PO4, 22.3 mg / L MnSO4·4H2O, 8.6 mg / L ZnSO4·7H2O, 6.2 mg / L H3BO3, 0.83 mg / L KI, 0.25 mg / L Na2MoO4·2H2O, 0.25 mg / L CuSO4·5H2O, 0.025 mg / L CoCl2, 37.13 mg / L FeSO4·7H2O, 49.67 mg / L Na2·EDTA, 100 mg / L inositol, 2.0 mg / L glycine, 0.1 mg / L thiamine hydrochloride, 0.5 mg / L nicotinic acid, 0.5 mg / L pyridoxine hydrochloride, pH 5.8, and the solvent is water. 1 / 2LS means that the dosages of NH4NO3, KNO3, CaCl2·2H2O, MgSO4·7H2O, and KH2PO4 are halved, and the others remain unchanged.

[0035] (6) Cultivation:

[0036] S1. Cultivation site: Use a modern greenhouse with a cooling device. Cover the inside of the greenhouse roof with a shading net, and cover the outside and around of the roof with a 50% shading black gauze net. Equip the greenhouse with an automatic spraying facility for water spraying and moisturizing and fertilization. Divide the greenhouse into pools of about 20 m 2 in size, with a pool height of 50 cm. Each pool needs to be equipped with a water inlet and a water outlet for regular water change. Soak and disinfect with potassium permanganate at a ratio of 1:10000 for 2 h one day before use, and then rinse with clean water once.

[0037] S2. Planting: Wash the tissue culture seedlings with tap water. Cut the rock wool into strips with the same width as the height of the planting basket, soak it with clean water, wrap the roots of the tissue culture seedlings in the rock wool, and add about 3 g of slow-release fertilizer (Osmocote, No. 1, N:P:K = 14:14:14). Roll it into a cylindrical shape. The diameter of the rolled rock wool cylinder is the same as the diameter of the planting basket, and place it in the planting basket. The bottom of the rock wool is closely attached to the bottom of the planting basket. Place the planted Anubias seedlings in a tray, and place the tray on the cultivation bed for cultivation. The water level should be higher than the leaves of the Anubias tissue culture seedlings. After new leaves grow, lower the water level to be flush with the planting basket.

[0038] S3. Management

[0039] S3.1. Water management: Replace the water in the cultivation pool once a week. Spray water regularly every day for leaf moisturizing. From 8:00 to 18:00 in summer and autumn, spray water once every 1 - 1.5 h, and the spraying time is about 1 - 3 min, just wetting the entire leaf surface; from 9:00 to 17:00 in winter and spring, spray water once every 1.5 - 2 h, and the spraying time is about 1 - 3 min, just wetting the entire leaf surface. The pH of the water in the cultivation pool should be controlled at 5.5 - 7.0. If it is too high or too low, replace the water.

[0040] S3.2. Fertilizer management: After the first new leaf of the Anubias seedlings grows, spray with 1000 - fold compound fertilizer (Yara, Norway, N:P:K = 15:15:15) + 20000 - fold urea solution, and the spraying time for each time is 2 - 5 min. Then fertilize once every half month.

[0041] S3.3. Temperature management: It can grow at 15 - 35 °C, and the suitable temperature is 18 - 30 °C. When the temperature is lower than 15 °C, pull down all the plastic films around the greenhouse for heat preservation. When the temperature is lower than 5 °C, the heating facilities can be turned on if conditions permit.

[0042] Comparative Example 1:

[0043] Except for the following different operations, the rest of the operations in this comparative example are the same as those in the example and will not be elaborated.

[0044] (1) Explant preparation: Select healthy and pest - free Anubias stem tips, remove the outer leaves, wash the surface impurities with tap water, soak them in washing powder for 30 min, and then wash them clean with tap water.

[0045] (2) Explant disinfection: Place the explants in a laminar flow hood. First, disinfect them with 75% alcohol for 30 s - 1 min, wash them 3 times with sterile water, then disinfect them with 0.1% mercuric chloride for 28 min, shake constantly during this period, and finally wash them 5 times with sterile water.

[0046] (3) Induction of adventitious buds: After the disinfected explants are peeled to 1 mm 3Shoot tips of about the size were inoculated into a medium of MS + 4 mg / L 6-BA + 0.1 mg / L NAA + 30 g / L sucrose + 5.5 g / L carrageenan. After inoculation, they were placed in a culture room for dark culture at a temperature of 25 ± 2°C.

[0047] Comparative Example 2:

[0048] Except for the following different operations, the remaining operations of this comparative example were the same as those of the example and will not be elaborated.

[0049] (1) Explant preparation: Healthy water fern stem segments without pests and diseases were selected. After washing off the surface impurities with tap water, they were soaked in washing powder for 30 min and then washed clean with tap water. Axillary buds of the water fern were cut with a surgical blade, and the length of the axillary buds was 3 - 5 mm.

[0050] (2) Explant disinfection: The explants were placed in a laminar flow hood. First, they were disinfected with 75% alcohol for 30 s - 1 min, washed 3 times with sterile water, then disinfected with 0.1% mercuric chloride for 24 min with continuous shaking during this period, and finally washed 5 times with sterile water.

[0051] (3) Induction of adventitious buds: After disinfection, the explants were peeled to obtain shoot tips of about 1 mm 3 in size and inoculated into a medium of MS + 4 mg / L 6-BA + 0.1 mg / L NAA + 30 g / L sucrose + 5.5 g / L carrageenan. After inoculation, they were placed in a culture room for dark culture at a temperature of 25 ± 2°C.

[0052] Comparative Example 3:

[0053] Except for the following different operations, the remaining operations of this comparative example were the same as those of the example and will not be elaborated.

[0054] (3) Induction of adventitious buds: The parts of the explants that had come into contact with the disinfectant at both ends were cut off after disinfection, and then the explants were cut into thin slices about 1 mm thick and inoculated into a medium of MS + 4 mg / L 6-BA + 0.1 mg / L NAA + 3 mg / L inositol + 30 g / L sucrose + 5.5 g / L carrageenan. After inoculation, they were placed in a culture room for dark culture at a temperature of 25 ± 2°C.

[0055] (4) Proliferation culture: The induced adventitious bud clusters were cut into small clusters of 1 cm 2 in size and inoculated into a medium of MS + 3 mg / L 6-BA + 0.1 mg / L NAA + 30 g / L sucrose + 5.5 g / L carrageenan. After inoculation, they were placed in a culture room for culture at a temperature of 25 ± 2°C, with dark culture for the first 3 days and then 6 h of light per day at a light intensity of 1000 lux.

[0056] (5) Rooting culture: Cut the proliferated buds with good growth and 2 leaves showing into single plants and inoculate them into the rooting medium, which is: 1 / 2MS + 0.1 mg / L NAA + 30 g / L sucrose + 5.5 g / L carrageenan + 0.5 g / L activated carbon. After inoculation, place them in the culture room for cultivation. The cultivation temperature is 25 ± 2 °C, with dark cultivation for the first 3 days, and then 10 hours of light per day with a light intensity of 2000 lux. 1 / 2MS means that the dosage of macronutrients in the MS medium is halved, and the others remain unchanged.

[0057] Comparative Example 4:

[0058] Except for the following different operations, the remaining operations of this comparative example are the same as those of the examples and will not be elaborated.

[0059] (3) Induction of adventitious buds: Cut off the parts of the explants in contact with the disinfectant at both ends after disinfection, and then cut them into thin slices with a thickness of about 1 mm, and inoculate them into the medium of MS + 4 mg / L 6-BA + 0.1 mg / L NAA + 30 g / L sucrose + 5.5 g / L carrageenan. After inoculation, place them in the culture room for dark cultivation, and the cultivation temperature is 25 ± 2 °C.

[0060] (4) Proliferation culture: Cut the induced adventitious bud clusters into small clusters of 1 cm 2 , and inoculate them into the medium of MS + 3 mg / L 6-BA + 0.1 mg / L NAA + 30 g / L sucrose + 5.5 g / L carrageenan. After inoculation, place them in the culture room for cultivation. The cultivation temperature is 25 ± 2 °C, with dark cultivation for the first 3 days, and then 6 hours of light per day with a light intensity of 1000 lux.

[0061] (5) Rooting culture: Cut the proliferated buds with good growth and 2 leaves showing into single plants and inoculate them into the rooting medium, which is: 1 / 2MS + 0.1 mg / L NAA + 30 g / L sucrose + 5.5 g / L carrageenan + 0.5 g / L activated carbon. After inoculation, place them in the culture room for cultivation. The cultivation temperature is 25 ± 2 °C, with dark cultivation for the first 3 days, and then 10 hours of light per day with a light intensity of 2000 lux.

[0062] Table 1

[0063]

[0064]

[0065] Statistically analyze the contamination rate, browning rate, disinfection success rate (i.e., explant survival rate), bud induction time, subculture cycle, proliferation coefficient, and rooting time of the examples and Comparative Examples 1 - 4, and the results are shown in Table 1.

[0066] In the present invention, the young flower stalks that have not yet emerged from the leaf sheaths are used as explants. The success rate of disinfection with mercuric chloride for 8 minutes can reach 96.74%, greatly shortening the disinfection time of mercuric chloride, minimizing the harm of heavy metals to the explants to the greatest extent, and having a fast growth recovery, a short subculture cycle, a high proliferation coefficient, and a short rooting time.

[0067] Comparative Example 5:

[0068] Except for the following different operations, the remaining operations of this comparative example are the same as those of the embodiment and will not be elaborated.

[0069] (6) Cultivation:

[0070] S1. Cultivation site: Use a modern greenhouse with a cooling device. The inside of the greenhouse roof is covered with a shading net, and a layer of black gauze net with a shading degree of 50% is covered outside and around the roof. Automatic spraying facilities are equipped in the greenhouse for spraying water to maintain humidity and fertilization. Divide it into pools of about 20 m 2 or so, with a pool height of 50 cm. Each pool needs to be equipped with a water inlet and a water outlet for regular water replacement. Soak and disinfect with 1:10,000 potassium permanganate for 2 h one day before use, and then rinse with clean water once.

[0071] S2. Planting: Use a disposable plastic cup with a height of 3 - 5 cm and holes at the bottom as the planting container. Fill the waterweed mud to 3 / 4 of the cup, then bury the roots of the cleaned water fern tissue culture seedlings into the waterweed mud. Finally, place the planted water fern seedlings on a tray and place the tray on the cultivation bed for cultivation. The water level should be higher than the leaves of the water fern tissue culture seedlings. After new leaves grow, lower the water level to be flush with the planting container.

[0072] S3. Management

[0073] S3.1. Water management: The water in the cultivation pool is changed once a week, and the spray is regularly sprayed every day for leaf moisturization. From 8:00 to 18:00 in summer and autumn, spray water once every 1 - 1.5 h, and the spraying time is about 1 - 3 min, so that the entire leaf surface is moist; from 9:00 to 17:00 in winter and spring, spray water once every 1.5 - 2 h, and the spraying time is about 1 - 3 min, so that the entire leaf surface is moist. The pH of the water in the cultivation pool is preferably controlled at 5.5 - 7.0. If it is too high or too low, change the water.

[0074] S3.2. Fertilizer management: After the water fern seedlings grow the first new leaf, spray with a 1000-fold liquid compound fertilizer (N:P:K = 15:15:15) + a 20,000-fold liquid urea, and the spraying time for each time is 2 - 5 min. Then fertilize once every half month.

[0075] S3.3. Temperature management: It can grow at 15 - 35°C, and the suitable temperature is 18 - 30°C. When the temperature is lower than 15°C, the plastic films around the greenhouse can be fully pulled down for heat preservation. When the temperature is lower than 5°C, the heating facilities can be turned on if available.

[0076] Comparative Example 6:

[0077] Except for the following different operations, the remaining operations of this comparative example are the same as those of the example and will not be elaborated.

[0078] (6) Cultivation:

[0079] S1. Cultivation site: Use a modern greenhouse with cooling equipment. The inside of the greenhouse roof is covered with a shading net, and a layer of black gauze net with a shading degree of 50% is covered on the outside and around the roof. Automatic spraying facilities are equipped inside the greenhouse for spraying water to maintain humidity and for fertilization. Ridges are formed with river sand according to the situation of the greenhouse, generally 8m long, 2m wide, and 6 - 8cm high.

[0080] S2. Planting: After cleaning the root medium of the tissue - cultured Anubias seedlings, bury the roots in the river sand.

[0081] S3. Management

[0082] S3.1. Water management: Spray regularly every day for leaf moisturization. From August to October and from March to May, spray water once every 1 - 1.5 hours from 8:00 to 18:00, and the spraying time is about 1 - 3 minutes, making sure the entire leaf surface is wet; from November to February and from June to July, spray water once every 1.5 - 2 hours from 9:00 to 17:00, and the spraying time is about 1 - 3 minutes, making sure the entire leaf surface is wet.

[0083] S3.2. Fertilizer management: After the Anubias seedlings grow the first new leaf, spray with a 1000 - fold liquid compound fertilizer (N:P:K = 15:15:15) + a 20000 - fold liquid urea, and the spraying time for each time is 2 - 5 minutes. Then fertilize once every half month.

[0084] S3.3. Temperature management: It can grow at 15 - 35°C, and the suitable temperature is 18 - 30°C. When the temperature is lower than 15°C, the plastic films around the greenhouse can be fully pulled down for heat preservation. When the temperature is lower than 5°C, the heating facilities can be turned on if available.

[0085] Comparative Example 7:

[0086] Except for the following different operations, the remaining operations of this comparative example are the same as those of the example and will not be elaborated.

[0087] (6) Cultivation

[0088] S1. Cleaning: Wash off the root medium with clean water and separate the seedlings into single plants;

[0089] S2. Planting: Plant in river sand or black ceramic granule substrate with a diameter of 2 - 4 mm that has been cleaned in an aquarium, and then add tap water to the aquarium until it is about 1 cm from the top of the tank.

[0090] S3. Maintenance and management: The filtration used in the aquarium is a brand - new filtration device or an old device that has been cleaned. Provide 6 - 10 hours of light per day with a light intensity of about 5000 lux; control the temperature at about 25 °C. If the temperature is higher than 27 °C or lower than 20 °C, turn on the indoor air conditioner for temperature control, or use a special cold water machine or heating rod for the fish tank for temperature control; add CO2 until bubbles can be produced after the aquarium plants are illuminated; change the water once a week, and the amount of water changed each time is 1 / 4 - 1 / 3 of the total water volume of the tank. Insert a slow - release root fertilizer rod for aquatic plants (Cuihu Aquatic Plant Cultivation Research Institute) to the bottom of the aquarium substrate every 2 - 3 months, 15×15 cm 2 Insert 1 piece.

[0091] Table 2

[0092]

[0093]

[0094] Count the growth time, survival rate, and whether cup replacement is needed for the examples and comparative examples 5 - 7. The results are shown in Table 2.

[0095] For the cultivation method of the present invention, for medium - large water ferns such as Batrachospermum, Acrostichum speciosum, A. aureum, and A. danaeifolium, they can be sold after 240 days of cultivation; for small water ferns such as Microsorum pteropus 'Minima', Ficus microcarpa 'Golden Leaves', F. microcarpa 'Nitida', F. microcarpa 'King', and F. microcarpa 'Peony', they can be sold after 150 days of cultivation. The method of the present invention has a short growth time, a low incidence rate, and is easy to operate.

[0096] The above - mentioned examples are the preferred embodiments of the present invention, but the embodiments of the present invention are only used to illustrate the technical solutions of the present invention rather than to limit. Within the scope defined by the claims of the present invention, any modifications and changes made by technicians in the art to the present invention should also fall within the protection scope of the claims of the present invention.

Claims

1. A method for the propagation and cultivation of Anubias, characterized in that, It includes the following steps: Select the young flower stalks of Anubias that have not yet emerged from the leaf sheaths. After disinfection, remove the parts at both ends that have come into contact with the disinfectant. Cut them into thin slices and inoculate them on the adventitious bud medium LS + 3 - 5 mg / L 6 - BA + 0.1 - 0.2 mg / L NAA + 2 - 4 mg / L inositol + 29 - 31 g / L sucrose + 5 - 6 g / L carrageenan, and conduct adventitious bud induction in the dark. Cut the adventitious buds into small clusters and inoculate them on the proliferation medium LS + 2 - 4 mg / L 6 - BA + 0.1 - 0.2 mg / L NAA + 29 - 31 g / L sucrose + 5 - 6 g / L carrageenan for proliferation culture. Cut the proliferated buds with leaves into single plants and inoculate them on the rooting medium 1 / 2LS + 0.1 - 0.2 mg / L NAA + 29 - 31 g / L sucrose + 5 - 6 g / L carrageenan + 0.5 - 1 g / L activated carbon, and obtain tissue - cultured seedlings through rooting culture. After washing the roots of the tissue - cultured seedlings, wrap them with rock wool, fix the rock wool, add water to submerge the leaves of the tissue - cultured seedlings, and lower the water level after new leaves grow; conduct water and fertilizer management; The components of the LS medium are as follows: 1650 mg / L NH4NO3, 1500 mg / L KNO3, 480 mg / L CaCl2·2H2O, 370 mg / L MgSO4·7H2O, 227 mg / L KH2PO4, 22.3 mg / L MnSO4·4H2O, 8.6 mg / L ZnSO4·7H2O, 6.2 mg / L H3BO3, 0.83 mg / L KI, 0.25 mg / L Na2MoO4·2H2O, 0.25 mg / L CuSO4·5H2O, 0.025 mg / L CoCl2, 37.13 mg / L FeSO4·7H2O, 49.67 mg / L Na2·EDTA, 100 mg / L inositol, 2.0 mg / L glycine, 0.1 mg / L thiamine hydrochloride, 0.5 mg / L nicotinic acid, 0.5 mg / L pyridoxine hydrochloride, pH 5.8, and the solvent is water.

2. The method according to claim 1, characterized in that, The adventitious bud medium is LS + 4 mg / L 6 - BA + 0.1 mg / L NAA + 3 mg / L inositol + 30 g / L sucrose + 5.5 g / L carrageenan.

3. The method according to claim 1, characterized in that The proliferation medium is LS + 3 mg / L 6 - BA + 0.1 mg / L NAA + 30 g / L sucrose + 5.5 g / L carrageenan.

4. The method according to claim 1, wherein The rooting medium is 1 / 2LS + 0.1 mg / L NAA + 30 g / L sucrose + 5.5 g / L carrageenan + 0.5 g / L activated carbon.

5. The method according to claim 1, characterized in that, The thickness of the thin slices is 1 - 2 mm.

6. The method according to claim 1, wherein The disinfection method is: After washing the impurities on the surface of the explant with tap water, place it in a laminar flow hood. First, disinfect it with 75% alcohol for 30 s - 1 min, wash it 3 times with sterile water, then disinfect it with 0.1% mercuric chloride for 8 min, shake it constantly during this period, and finally wash it 5 times with sterile water.

7. The method according to claim 1, wherein When wrapping with rock wool, add 2 - 4 g of slow - release fertilizer (Osmocote); the fixation of the rock wool is to roll the rock wool into a cylindrical shape. The diameter of the rolled rock wool cylinder is the same as the diameter of the planting basket, place it in the planting basket, and the bottom of the rock wool is closely attached to the bottom of the planting basket.

8. The method according to claim 1, wherein The water and fertilizer management is carried out by using an automatic spraying facility. In summer and autumn, water is sprayed once every 1 - 1.5 hours from 8:00 to 18:00, and the spraying time is 1 - 3 minutes until the entire leaf surface is wet. In winter and spring, water is sprayed once every 1.5 - 2 hours from 9:00 to 17:00, and the spraying time is 1 - 3 minutes until the entire leaf surface is wet. After the first new leaf grows on the tissue-cultured seedling, spraying is started with a 1000-fold liquid of compound fertilizer and a 20000-fold liquid of urea, and the spraying time for each time is 2 - 5 minutes, once every half month; the compound fertilizer has an elemental mass ratio of N:P:K = 15:15:

15.

9. The method according to claim 1, wherein The water ferns include **B. bartlettii**, **F. acuminata**, **F. heteroclita** and **F. deltoidea**. Notes: For the scientific names of plants in , it is assumed that the Chinese names you provided are specific plant names, and I have translated them into the corresponding scientific names according to common knowledge. If there are inaccuracies, please adjust according to the actual situation.

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