Efficient asexual propagation method for ficus pandurata applicable to simple conditions

Abstract

The invention provides an efficient asexual propagation method of ficus pandurata suitable for simple condition application. The method comprises the following steps: (1) explant disinfection: soaking a stem with an axillary bud in a composite disinfectant containing mercuric chloride; (2) primary culture: inoculating the disinfected stem into a starting culture medium containing cytokinin and auxin for culture; (3) multiplication culture: transferring the callus formed by primary culture to a differential culture medium to induce cluster buds; (4) carrying out vector rooting, namely cutting robust buds obtained by multiplication culture, and inserting the robust buds into a rooting culture medium containing microencapsulated auxin; and (5) transplanting and hardening seedlings: treating the tissue culture seedlings with a protective solution, and transplanting the treated tissue culture seedlings to a compound matrix. The method can realize substantive transformation from a specialized tissue culture technology to basic-level production, and solves the industrialization bottlenecks such as equipment threshold and operation complexity.

Description

Technical Field

[0001] This invention belongs to the field of plant asexual reproduction technology, specifically relating to a highly efficient asexual reproduction method for Ficus lyrata suitable for simple applications. Background Technology

[0002] The annual demand for Ficus lyrata bonsai in the indoor green plant market remains stable at over 1.5 million pots. However, traditional propagation by cuttings has long suffered from three core pain points: First, the survival rate fluctuates wildly. When farmers use open-air sand beds for cuttings, the rooting rate is often less than 30% during the rainy season, mainly because rainwater carries pathogens that cause the cuts to rot. Although humidity can be maintained by artificial spraying during the dry season, water and electricity costs will increase by 40%. Second, the consumption of mother plant resources is too large. Each high-quality mother tree can only provide 8 to 10 qualified cuttings at a time. Large-scale production requires stockpiling thousands of mother plants, and each acre can only support the production needs of 3,000 seedlings. Third, existing tissue culture technology is difficult to implement. Although laboratory results show that the propagation coefficient of tissue culture seedlings can reach more than 6 times, it depends on equipment such as clean benches and light incubators. The equipment penetration rate of county-level nurseries is less than 15%, and the purchase cost of a single clean bench exceeds 50,000 yuan.

[0003] Existing improved technologies still have significant drawbacks. For example, a certain layering patent technology uses high-pressure sterilized moss to wrap branches, which shortens the rooting cycle to about 25 days, but the yield per mother plant is only 3 to 5 seedlings, limiting the increase in production capacity. Another example is a two-step tissue culture protocol disclosed in a certain literature, which first induces callus tissue with 2.0 mg / L cytokinin BA, then transfers it with 0.5 mg / L BA to promote bud differentiation, and finally roots it on a 0.2 mg / L auxin IBA medium. This method has a high propagation coefficient in laboratory environments, but its practical application has exposed three fatal problems: First, the explant sterilization process requires precise control of 0.1% mercuric chloride for 10 minutes; a time error of more than 15 seconds will lead to browning and death of the material, with an error rate of over 50% for frontline operators; second, the cumulative contamination rate of the three transfer operations is as high as 37%; and finally, three different culture media formulations are required, with labor costs accounting for 60% of the seedling price.

[0004] These realities expose a core contradiction: the high technical barriers of professional tissue culture cannot be matched with the simplified needs of grassroots production. Therefore, there is an urgent need for a method of asexual propagation of Ficus fragrans that can achieve high survival rates and low pollution rates without relying on specialized equipment. Summary of the Invention

[0005] To overcome the shortcomings of existing technologies, a highly efficient asexual propagation method for Ficus lyrata suitable for simple applications is provided.

[0006] To achieve the above objectives, the present invention provides the following technical solution: A highly efficient asexual propagation method for Ficus lyrata suitable for simple applications, comprising the following steps: (1) Disinfection of explants: Soak stem segments with axillary buds in a compound disinfectant solution containing mercuric chloride; (2) Primary culture: After disinfection, the stem segments were inoculated into a starter culture medium containing cytokinins and auxins; (3) Proliferation culture: The callus tissue formed by the primary culture was transferred to the differentiation medium to induce shoot clusters; (4) Carrier-based rooting: Healthy shoots obtained from proliferation culture are cut and inserted into a rooting medium containing microencapsulated auxin; (5) Transplanting and hardening off: After treatment with protective solution, tissue culture seedlings are transplanted to the compound substrate.

[0007] The specific steps for disinfecting the explants are as follows: Select semi-lignified branches at the top of healthy mother plants, retain two nodes with axillary buds, cut them into 4 cm long segments, immerse the cut stem segments in a compound disinfectant solution for 9-11 minutes, and simultaneously shake the container at a speed of 25-30 rpm.

[0008] The compound disinfectant comprises the following components by weight: 1000 parts distilled water, 0.8-1 parts mercuric chloride, 0.4-0.6 parts Tween 80 surfactant, 0.1-0.15 parts L-ascorbic acid, and 0.3-0.4 parts sodium citrate.

[0009] In the initial culture, the starter culture medium, by weight, includes: 35-40 parts MS medium base salt, 12-16 parts modified coconut milk powder, 8-12 parts trehalose, 6-8 parts agar, 1.9-2.1 parts cytokinin, 0.08-0.12 parts auxin, and 900 parts distilled water.

[0010] The cytokinin is 6-benzylaminopurine, and the auxin is α-naphthaleneacetic acid.

[0011] The modified coconut milk powder is prepared by centrifuging fresh coconut milk to remove the fat layer, adding pectinase equivalent to 1% of the weight of coconut milk, enzymatically hydrolyzing in a constant temperature water bath at 40-45℃ for 30-50 minutes, then mixing the enzymatic hydrolysate with palygorskite powder with a particle size of less than 5 micrometers at a mass ratio of 10-15:1 for adsorption, and drying to obtain the modified coconut milk powder.

[0012] In the proliferation culture, when the diameter of the callus tissue formed in the primary culture reaches 2.5-3.5 mm, it is transferred to the differentiation medium to induce shoot clusters; the differentiation medium, by weight, includes: 35-40 parts MS medium base salt, 10-15 parts trehalose, 5-10 parts agar, 0.8-1 parts 6-benzylaminopurine, and 0.10-0.15 parts α-naphthaleneacetic acid.

[0013] In the carrier-based rooting process, 200-300 microencapsulated auxin particles are mixed into each liter of rooting medium. The microencapsulated auxin is composed of indolebutyric acid and α-naphthaleneacetic acid loaded on a diatomaceous earth carrier. The mass ratio of indolebutyric acid to α-naphthaleneacetic acid is 2.8-3.2:1.

[0014] The preparation method of microencapsulated growth hormone is as follows: diatomaceous earth is calcined at 450-500℃ for 2 hours and then cooled; it is then immersed in a 65% ethanol solution containing 25 mg / mL indolebutyric acid and 8.3 mg / mL α-naphthaleneacetic acid for 1.5 hours; after vacuum drying at 60℃, it is passed through a 400-mesh sieve to obtain microencapsulated growth hormone.

[0015] The specific steps for transplanting and hardening off seedlings are as follows: After 18 days of rooting on the carrier, take out the tissue culture seedlings with well-developed root systems, wash off the agar from the roots, soak the roots in a 550-fold diluted solution of mancozeb and chitosan for 5 minutes, and transplant them into the compound substrate. The volume ratio of pine bark, vermiculite and biochar in the compound substrate is (4-4.5):(2-2.2):1.

[0016] Compared with the prior art, the advantages and beneficial effects of the present invention are as follows: 1. This invention significantly reduces the risk of explant browning through the synergistic effect of mercuric chloride and L-ascorbic acid in a compound disinfectant. While mercuric chloride provides broad-spectrum sterilization capabilities, L-ascorbic acid, as an antioxidant, can neutralize oxidative stress products at the incision site. This dual mechanism extends the disinfection time window from the rigid control of existing technologies to a flexible range of 9 to 11 minutes, greatly reducing the error rate of primary care procedures.

[0017] 2. In the initial culture stage, this invention improves callus induction efficiency through modified coconut milk powder. After pectin hydrolysis, coconut milk releases endogenous growth factors, which are then concentrated by the high specific surface area of ​​palygorskite powder. This modification allows the initiation culture medium to maintain stable cell division activity even under natural diffused light conditions. Furthermore, 6-benzylaminopurine and α-naphthaleneacetic acid form a hormonal balance at a specific concentration ratio, preventing excessive callus proliferation and laying the morphological foundation for subsequent bud differentiation.

[0018] 3. The porous structure formed by high-temperature calcination of the diatomaceous earth of this invention can efficiently load the complex auxin of indolebutyric acid and α-naphthaleneacetic acid. When embedded in the rooting medium, water gradually dissolves and releases auxin molecules through the microporous channels. This continuous supply mode simulates the physiological distribution gradient of endogenous hormones in plants, which is more conducive to the directional development of root primordia compared with traditional liquid addition methods.

[0019] 4. Regarding protective measures during the transplanting and hardening-off stage, this invention forms a dual physical-chemical protective layer by combining mancozeb and chitosan. Mancozeb forms an antibacterial film on the root surface to inhibit soil-borne disease infection, while chitosan's film-forming properties reduce water transpiration stress. This protective mechanism, combined with the good permeability of the pine bark-vermiculite-biochar substrate, creates a synergistic effect, enabling tissue-cultured seedlings to quickly adapt to a non-sterile environment. Detailed Implementation

[0020] The technical solutions in the embodiments of the present invention will be clearly and completely described below. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0021] In the specific embodiments of this application, the sources of various raw materials are briefly described as follows: Mercuric chloride: purchased from Sinopharm Chemical Reagent Co., Ltd., the product is of analytical grade, CAS number 7487-94-7.

[0022] Tween 80 surfactant: purchased from Nanjing Chemical Reagent Co., Ltd., CP grade, CAS No. 9005-65-6.

[0023] L-Ascorbic acid: Purchased from TCI (Shanghai) Chemical Industry Development Co., Ltd., product code: A0537.

[0024] Sodium citrate: purchased from Henan Juteng Chemical Products Co., Ltd., CAS No. 77-92-9.

[0025] MS medium base salts: purchased from Pursray (Shanghai) Biopharmaceutical Co., Ltd. This product does not contain agar or sucrose.

[0026] α-Naphthaleneacetic acid: purchased from Shanghai Yuanye Biotechnology Co., Ltd., CAS No. 86-87-3, purity ≥98%.

[0027] 6-Benzylaminopurine: Purchased from Nanjing Chemical Reagent Co., Ltd., CAS 1214-39-7, active ingredient content ≥98%.

[0028] Pectinase: Purchased from Xiasheng (Beijing) Biotechnology Development Co., Ltd., with an enzyme activity ≥4500 U / g.

[0029] Pallas's stone powder: purchased from Lingshou County Zhanteng Mineral Products Processing Plant.

[0030] Indolebutyric acid: purchased from Jinan Hengyuan Chemical Co., Ltd., CAS No. 133-32-4, plant culture grade.

[0031] Agar: Purchased from Fujian Huanke Biotechnology Co., Ltd., the product is strip agar for plant tissue culture.

[0032] Trehalose: Purchased from Shandong Freda Biotechnology Co., Ltd., CAS No. 99-20-7.

[0033] Diatomaceous earth: purchased from Chengdu Dinghengda Chemical Products Co., Ltd.

[0034] Chitosan: Purchased from Shandong Huian Chemical Co., Ltd., CAS No. 9012-76-4.

[0035] Manganese zinc: purchased from Limin Chemical Co., Ltd., CAS No. 8018-01-7.

[0036] The technical solution of this application is as follows: A highly efficient asexual propagation method for Ficus lyrata suitable for simple applications, comprising the following steps: (1) Disinfection of explants: Soak stem segments with axillary buds in a compound disinfectant solution containing mercuric chloride; (2) Primary culture: After disinfection, the stem segments were inoculated into a starter culture medium containing cytokinins and auxins; (3) Proliferation culture: The callus tissue formed by the primary culture was transferred to the differentiation medium to induce shoot clusters; (4) Carrier-based rooting: Healthy shoots obtained from proliferation culture are cut and inserted into a rooting medium containing microencapsulated auxin; (5) Transplanting and hardening off: After treatment with protective solution, tissue culture seedlings are transplanted to the compound substrate.

[0037] The specific steps for disinfecting the explants are as follows: Select semi-lignified branches at the top of healthy mother plants, retain two nodes with axillary buds, cut them into 4 cm long segments, immerse the cut stem segments in a compound disinfectant solution for 9-11 minutes, and simultaneously shake the container at a speed of 25-30 rpm.

[0038] The compound disinfectant comprises the following components by weight: 1000 parts distilled water, 0.8-1 parts mercuric chloride, 0.4-0.6 parts Tween 80 surfactant, 0.1-0.15 parts L-ascorbic acid, and 0.3-0.4 parts sodium citrate.

[0039] In the initial culture, the starter culture medium, by weight, includes: 35-40 parts MS medium base salt, 12-16 parts modified coconut milk powder, 8-12 parts trehalose, 6-8 parts agar, 1.9-2.1 parts cytokinin, 0.08-0.12 parts auxin, and 900 parts distilled water.

[0040] The cytokinin is 6-benzylaminopurine, and the auxin is α-naphthaleneacetic acid.

[0041] The modified coconut milk powder is prepared by centrifuging fresh coconut milk to remove the fat layer, adding pectinase equivalent to 1% of the weight of coconut milk, enzymatically hydrolyzing in a constant temperature water bath at 40-45℃ for 30-50 minutes, then mixing the enzymatic hydrolysate with palygorskite powder with a particle size of less than 5 micrometers at a mass ratio of 10-15:1 for adsorption, and drying to obtain the modified coconut milk powder.

[0042] In the proliferation culture, when the diameter of the callus tissue formed in the primary culture reaches 2.5-3.5 mm, it is transferred to the differentiation medium to induce shoot clusters; the differentiation medium, by weight, includes: 35-40 parts MS medium base salt, 10-15 parts trehalose, 5-10 parts agar, 0.8-1 parts 6-benzylaminopurine, and 0.10-0.15 parts α-naphthaleneacetic acid.

[0043] In the carrier-based rooting process, 200-300 microencapsulated auxin particles are mixed into each liter of rooting medium. The microencapsulated auxin is composed of indolebutyric acid and α-naphthaleneacetic acid loaded on a diatomaceous earth carrier. The mass ratio of indolebutyric acid to α-naphthaleneacetic acid is 2.8-3.2:1.

[0044] The preparation method of microencapsulated growth hormone is as follows: diatomaceous earth is calcined at 450-500℃ for 2 hours and then cooled; it is then immersed in a 65% ethanol solution containing 25 mg / mL indolebutyric acid and 8.3 mg / mL α-naphthaleneacetic acid for 1.5 hours; after vacuum drying at 60℃, it is passed through a 400-mesh sieve to obtain microencapsulated growth hormone.

[0045] The specific steps for transplanting and hardening off seedlings are as follows: After 18 days of rooting on the carrier, take out the tissue culture seedlings with well-developed root systems, wash off the agar from the roots, soak the roots in a 550-fold diluted solution of mancozeb and chitosan for 5 minutes, and transplant them into the compound substrate. The volume ratio of pine bark, vermiculite and biochar in the compound substrate is (4-4.5):(2-2.2):1.

[0046] In this application, 6-benzylaminopurine is abbreviated as BA, auxin is abbreviated as NAA (α-naphthaleneacetic acid), and indolebutyric acid is abbreviated as IBA.

[0047] The greatest systemic advantage of this invention lies in the optimized design of the process integration. From the disinfection and anti-browning mechanism, the physiological regulation of callus induction, the slow-release control of root development, to the film-forming technology for transplant protection, the technical features of each link form a closed-loop synergy. This design breaks through and transforms the key control points of professional tissue culture into simple operating steps, enabling the entire process to be carried out in ordinary plastic greenhouses, truly solving the core contradiction between equipment dependence and technology popularization.

[0048] The technical solutions of the present invention are further illustrated below through examples and comparative examples, but the scope of protection of the present invention is not limited thereto.

[0049] Example 1 Select semi-lignified branches from the top of healthy Ficus lyrata mother plants, retaining two nodes with axillary buds, and cut them into 4 cm stem segments. Immerse the stem segments in a compound disinfectant solution composed of 1000 parts distilled water, 0.8 parts mercuric chloride, 0.6 parts Tween 80 surfactant, 0.15 parts L-ascorbic acid, and 0.3 parts sodium citrate for 9 minutes, shaking the container at 30 rpm during the treatment. After disinfection, inoculate the stem segments into a starter culture medium containing 40 parts MS medium base salts, 16 parts modified coconut milk powder, 12 parts trehalose, 8 parts agar, 2.1 parts 6-benzylaminopurine, 0.12 parts α-naphthaleneacetic acid, and 900 parts distilled water. After culturing for 27 days under natural diffused light, the callus tissue was transferred to differentiation medium when it reached a diameter of 3.5 mm. This differentiation medium contained 40 parts MS medium base salts, 15 parts trehalose, 10 parts agar, 1 part 6-benzylaminopurine, and 0.15 parts α-naphthaleneacetic acid. After 19 days of proliferation culture, robust shoots with a height of 3.5 cm were excised and inserted into rooting medium containing microencapsulated auxin. 300 microencapsulated auxin capsules were mixed into each liter of medium. The microencapsulated auxin was prepared as follows: diatomaceous earth was calcined at 500°C for 2 hours, then immersed in a 65% (v / v) ethanol solution containing 25 mg / mL indolebutyric acid and 8.3 mg / mL α-naphthaleneacetic acid for 1.5 hours. After vacuum drying at 60°C, the solution was passed through a 400-mesh sieve. After 18 days of rooting culture, the tissue culture seedlings were removed, and the roots were soaked in a 550-fold dilution of mancozeb and chitosan for 5 minutes. They were then transplanted into a compound substrate with a volume ratio of pine bark, vermiculite and biochar of 4.5:2.2:1.

[0050] Example 2 In this embodiment, the similarities to those in Embodiment 1 will not be repeated, and the differences are as follows: The compound disinfectant solution consisted of 1000 parts distilled water, 1 part mercuric chloride, 0.4 parts Tween 80 surfactant, 0.1 parts L-ascorbic acid, and 0.4 parts sodium citrate. The soaking time was 11 minutes, and the shaking speed was 25 rpm. The starting medium was adjusted to contain 35 parts MS medium base salts, 12 parts modified coconut milk powder, 8 parts trehalose, 6 parts agar, 1.9 parts 6-benzylaminopurine, 0.08 parts α-naphthaleneacetic acid, and 900 parts distilled water. When the callus tissue formed in the primary culture reached a diameter of 2.5 mm, it was transferred to the differentiation medium, which contained 35 parts MS medium base salts, 10 parts trehalose, 7 parts agar, 0.9 parts 6-benzylaminopurine, and 0.12 parts α-naphthaleneacetic acid. 250 microencapsulated auxin particles were mixed into each liter of rooting medium, and the diatomaceous earth was calcined at 475 degrees Celsius during its preparation. The volume ratio of the transplanting substrate was adjusted to pine bark:vermiculite:biochar = 4.2:2.1:1.

[0051] Example 3 In this embodiment, the similarities to those in Embodiment 1 will not be repeated, and the differences are as follows: The compound disinfectant solution consisted of 1000 parts distilled water, 0.9 parts mercuric chloride, 0.5 parts Tween 80 surfactant, 0.12 parts L-ascorbic acid, and 0.35 parts sodium citrate. The soaking time was 10 minutes, and the shaking speed was 28 rpm. The starting medium contained 37 parts MS medium base salts, 14 parts modified coconut milk powder, 10 parts trehalose, 7 parts agar, 2.0 parts 6-benzylaminopurine, 0.10 parts α-naphthaleneacetic acid, and 900 parts distilled water. When the callus reached a diameter of 3.0 mm, it was transferred to the differentiation medium, which contained 38 parts MS medium base salts, 12 parts trehalose, 5 parts agar, 0.8 parts 6-benzylaminopurine, and 0.10 parts α-naphthaleneacetic acid. 200 microencapsulated auxin particles were mixed into each liter of rooting medium. The diatomaceous earth was calcined at 450°C. The volume ratio of the transplanting substrate was pine bark:vermiculite:biochar = 4:2:1.

[0052] Comparative Example 1 In this comparative example, the similarities with Example 1 will not be repeated, and the differences are as follows: Remove L-ascorbic acid from compound disinfectant solution.

[0053] Comparative Example 2 In this comparative example, the similarities with Example 2 will not be repeated, and the differences are as follows: Unmodified coconut milk powder was used as the starter culture medium.

[0054] Comparative Example 3 In this comparative example, the similarities with Example 3 will not be repeated, and the differences are as follows: Add auxin solution directly during the rooting stage.

[0055] Comparative Example 4 In this comparative example, the similarities with Example 1 will not be repeated, and the differences are as follows: Do not apply protective solution during transplanting.

[0056] Comparative Example 5 In this comparative example, the similarities with Example 1 will not be repeated, and the differences are as follows: The transplanting substrate was changed to ordinary garden soil.

[0057] Performance test results and analysis Fifty batches of repeated tests were conducted in a simple plastic greenhouse according to the parameters of the examples and comparative examples, respectively. The results of the core indicator tests are summarized in Table 1.

[0058] As shown in Table 1, the explant contamination rate in the example groups was below 5.1%, while that in Comparative Example 1 increased to 31.6%, confirming the core value of L-ascorbic acid in the compound disinfectant. When mercuric chloride kills surface microorganisms, L-ascorbic acid blocks the oxidation chain reaction of phenolic substances by scavenging free radicals, preventing browning of the explant incision site. The shaking operation improves the penetration efficiency of the disinfectant, extending the time tolerance window to the 9-11 minute range.

[0059] Comparative Example 2 showed a 24.7% decrease in the propagation coefficient due to the use of unmodified coconut milk powder, indicating a synergistic effect between the endogenous growth factors released by pectinase and the trace elements adsorbed by palygorskite. Electron microscopy revealed that the modified coconut milk powder treatment group exhibited a 2.3-fold increase in callus cell division rate, laying a material foundation for subsequent shoot differentiation.

[0060] Table 1 Test Results

[0061] Comparative Example 3 showed a 19.1% decrease in root development index, revealing the sustained-release value of microencapsulated auxin. The mesoporous structure formed by calcining diatomaceous earth at 450 to 500 degrees Celsius allows for the release of auxin at a gradient of 1.2 to 1.5 micrograms per day, precisely mimicking the vascular bundle hormone transport pattern and avoiding root tip malformation caused by excessively high local concentrations.

[0062] The survival rate of Comparative Example 4 dropped sharply by 18.8%, highlighting the membrane protection mechanism of the mancozeb-chitosan compound solution. Energy dispersive spectroscopy analysis showed that zinc covered 83% of the root surface, forming a physical antibacterial barrier, while chitosan sealed the intercellular spaces of epidermal cells through hydrogen bonding, reducing the transpiration rate by 42 percentage points.

[0063] The root development index of Comparative Example 5 decreased by 26.0%, which was attributed to the superior pore structure of the composite matrix. The mixture of pine bark, vermiculite, and biochar in a specific ratio achieved a porosity of 52.7%, significantly higher than the 31.4% of ordinary garden soil. This structure promoted gas exchange in the root hair zone, increasing mitochondrial activity in newly formed roots by 37 percentage points.

[0064] Test results show that the present invention forms a closed-loop process through the anti-browning synergistic mechanism of compound disinfectant, the enhancement of physiological activity of modified coconut milk powder, the slow-release root control technology of diatomaceous earth microcapsules, and the film-forming protective effect of transplant protection. While significantly reducing equipment dependence, it successfully constructs a highly efficient propagation system suitable for simple production environments. Its core breakthrough lies in transforming the key control points of professional tissue culture into operational units that can be implemented on a large scale.

[0065] The above description represents the preferred embodiments of the present invention. It should be noted that those skilled in the art can make various improvements and modifications without departing from the principles of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. A method for high efficient and asexual propagation of Ficus lyrata suitable for easy conditions, characterized in that, The method comprises the following steps: (1) Disinfecting the explant: immerse the stem section with axillary buds in a composite disinfectant solution containing mercuric chloride; (2) Primary culture: after disinfection, the stem section is inoculated into a starting culture medium containing cell division factor and auxin for culture; (3) Proliferation culture: the callus formed in the primary culture is transferred to a differentiation culture medium to induce cluster buds; (4) Carrierization and rooting: the healthy buds obtained in the proliferation culture are cut and inserted into a rooting culture medium containing microencapsulated auxin; (5) Transplanting and acclimatizing: the tissue culture seedlings are treated with a protective solution and then transplanted into a compound substrate.

2. A method for high efficient vegetative propagation of Ficus lyrata suitable for application under simple conditions according to claim 1, characterized in that, The specific steps for disinfecting the explant are as follows: select a healthy top semi-lignified branch of a mother plant, retain two nodes with axillary buds, cut into 4 cm length sections, and immerse the cut stem sections in a composite disinfectant solution for 9-11 minutes while oscillating the container at a speed of 25-30 rpm.

3. A method for high efficient vegetative propagation of Ficus lyrata suitable for application under simple conditions according to claim 2, characterized in that, The composite disinfectant solution comprises the following components in parts by mass: distilled water 1000 parts, mercuric chloride 0.8-1 part, Tween 80 surfactant 0.4-0.6 part, L-ascorbic acid 0.1-0.15 part, and sodium citrate 0.3-0.4 part.

4. A method for high efficient vegetative propagation of Ficus tikoua according to claim 1, wherein, In the primary culture, the starting culture medium comprises, in parts by mass: MS culture medium base salt 35-40 parts, modified coconut milk powder 12-16 parts, trehalose 8-12 parts, agar 6-8 parts, cell division factor 1.9-2.1 parts, auxin 0.08-0.12 parts, and distilled water 900 parts.

5. A method for high efficient vegetative propagation of Ficus tikoua according to claim 4, wherein, The cell division factor is 6-benzylaminopurine, and the auxin is alpha-naphthaleneacetic acid.

6. A method for high efficient vegetative propagation of Ficus tikoua according to claim 4, wherein, The preparation method of the modified coconut milk powder is as follows: remove the fat layer of fresh coconut milk by centrifugation, add pectinase equivalent to 1% of the weight of the coconut milk, and enzymatically hydrolyze in a constant-temperature water bath at 40-45°C for 30-50 minutes; then mix and adsorb the enzymatic hydrolysate with palygorskite powder with a particle size of less than 5 microns at a mass ratio of 10-15:1, and dry to obtain the modified coconut milk powder.

7. A method for high efficient and rapid propagation of Ficus tikoua Hance by tissue culture under simple conditions according to claim 1, characterized in that, In the proliferation culture, the callus formed in the primary culture is transferred to a differentiation culture medium to induce cluster buds when the diameter of the callus reaches 2.5-3.5 mm; the differentiation culture medium comprises, in parts by mass: MS culture medium base salt 35-40 parts, trehalose 10-15 parts, agar 5-10 parts, 6-benzylaminopurine 0.8-1 part, and alpha-naphthaleneacetic acid 0.10-0.15 part.

8. A method for high efficient vegetative propagation of Ficus tikoua according to claim 1, wherein, In the carrierization and rooting, 200-300 particles of microencapsulated auxin are mixed into each liter of rooting culture medium, and the microencapsulated auxin is composed of indolebutyric acid and alpha-naphthaleneacetic acid loaded on diatomite carriers; the mass ratio of the indolebutyric acid to the alpha-naphthaleneacetic acid is 2.8-3.2:

1.

9. A method for high efficient vegetative propagation of Ficus lyrata according to claim 8, wherein, The preparation method of the microencapsulated auxin is as follows: calcine diatomite at 450-500°C for 2 hours and then cool; immerse in a 65% ethanol solution containing 25 mg / mL of indolebutyric acid and 8.3 mg / mL of alpha-naphthaleneacetic acid for 1.5 hours; vacuum dry at 60°C and then sieve through a 400 mesh sieve to obtain the microencapsulated auxin.

10. A method for high efficient vegetative propagation of Ficus tikoua according to claim 1, wherein, The specific steps of the transplanting seedling training are as follows: after the carrierization rooting for 18 days, the tissue culture seedlings with developed root systems are taken out, the roots are washed clean, the roots are soaked in 550 times diluent of the compound of mancozeb and chitosan for 5 minutes, and then the roots are transplanted into the compound substrate, wherein the volume ratio of pine bark, vermiculite and biochar in the compound substrate is (4-4.5):(2-2.2):1.

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