Establishment method of rapid propagation system of Buddhida viridae

Abstract

The invention belongs to the technical field of plant tissue culture, and particularly discloses a method for establishing a rapid propagation system of Buddhist viridae. The method comprises the steps of explant disinfection, adventitious bud induction, bud proliferation and seedling strengthening and rooting, firstly, 75% alcohol is used for soaking for 30 seconds, then one drop of Tween-20 sodium hypochlorite solution with the concentration being 2% is added for disinfection for 15 minutes, the survival rate of explants reaches 73.3%, and the pollution rate is reduced to 11.1%; a specific hormone ratio is adopted, bud induction: 6-BA is 1.0 mg / L + IBA is 0.3 mg / L, and the germination rate is 94.4%; bud proliferation: 6-BA is 0.5 mg / L + IBA is 0.3 mg / L, and the proliferation coefficient is 4.1; and rooting: IBA is 0.2 mg / L and NAA is 0.2 mg / L, the rooting rate is 100%, the survival rate, the bud induction rate and the multiplication coefficient of the explants are remarkably improved, and the method is easy and convenient to operate, efficient, environmentally friendly and suitable for large-scale production.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of plant tissue culture, and particularly relates to a method for establishing a rapid propagation system of Philodendron'Florida Green'. BACKGROUND

[0002] Philodendron'Florida Green'has high ornamental value due to its unique leaf shape (similar to an airplane blade) and has a strong market demand. However, the traditional propagation by division is inefficient and has a long propagation cycle, and thus cannot meet the demand of large-scale production. Therefore, it is crucial to establish an efficient in vitro rapid propagation system for the commercial production of the variety.

[0003] At present, there are some studies on the tissue culture of Philodendron plants, such as Philodendron'Golden Diamond'and Philodendron'Hope', but there are still technical bottlenecks such as low survival rate of external sterilization, high contamination rate, low efficiency of bud induction and proliferation, and unstable rooting quality. In plant tissue culture, external sterilization is a key step that directly affects the success rate of subsequent culture. Traditional sterilization methods mostly use mercury chloride (HgCl2), which has good sterilization effect, but its high toxicity leads to high operation risk, and residual mercury ions are difficult to completely clean, which is harmful to the environment and human health. Low-toxicity disinfectants such as sodium hypochlorite (NaClO) and hydrogen peroxide (H2O2) are relatively safe, but they are easily washed away, have low sterilization effect, and can easily cause external explants to brown and die, with a low survival rate.

[0004] In addition, the existing bud induction medium for Philodendron plants mostly uses a combination of 6-BA (6-benzylaminopurine) and NAA (naphthalene acetic acid) or IBA (indole-3-butyric acid), but its effect has significant variety differences. 6-BA, as a cytokinin, promotes the initiation and proliferation of bud primordia by activating cyclins (such as Cyclin D), but too high a concentration of 6-BA can easily lead to vitrification or malformation of buds; NAA has high stability, but it can easily induce callus and inhibit direct differentiation of buds; IAA is easily affected by photolysis and enzymatic hydrolysis, and thus has poor stability, and therefore IBA is selected as an alternative to induce rooting and promote the robustness of buds.

[0005] At present, there is no report on a complete rapid propagation system for Philodendron'Florida Green', and the existing technology has problems such as low sterilization efficiency, low proliferation coefficient, and unstable rooting quality, especially lacking a scheme for establishing a rapid propagation system for Philodendron'Florida Green'by using high-efficiency sterilization and hormone regulation together. SUMMARY

[0006] The method optimizes the rapid propagation system of Florida Philodendron, establishes an efficient and low-toxicity disinfection scheme, and significantly reduces the pollution rate and improves the survival rate by using 75% alcohol soaking for 30s, and then adding 1 drop of Tween-20 in 2% sodium hypochlorite solution for disinfection for 15min, instead of mercury; the hormone ratio is used to improve the establishment of the rapid propagation system, the bud induction stage: 6-BA 1.0mg / L + IBA 0.3mg / L, the induction rate is as high as 94.4%, the bud proliferation stage: 6-BA 0.5mg / L + IBA 0.3mg / L, the proliferation coefficient is as high as 4.1, which is significantly higher than similar varieties; the rooting system adopts IBA 0.2mg / L + NAA 0.2mg / L combination, the rooting rate is 100%, the root system is strong, and the transplanting survival rate is high.

[0007] The application provides a method for establishing a rapid propagation system of Florida Philodendron, comprising the following steps: S1: external treatment and disinfection: irrigate the plants in advance, keep the potted soil dry, spray the plant leaves with carbendazim, select healthy and pest-free mother plants in the morning on sunny days, cut the strong growth stem segments with buds, wash the leaf surface of the stem segments with detergent, then rinse with running water, and then perform the following treatment on the cleaned stem segments in the clean bench: soak in 75% alcohol for 30s, shake constantly during the process, then rinse with sterile water for 3 times, then soak the stem segments in 2% sodium hypochlorite solution with 1 drop of Tween-20 for 15min, rinse with sterile water for 4-6 times after disinfection until no foam appears on the surface of the stem segments, place the disinfected stem segments on sterile filter paper to absorb water, cut the stem segments into wedge-shaped blocks with a bud point as the center, about 0.5cm-1.5cm long and 0.3cm-0.8cm thick using sterilized instruments, and make fresh wounds on three sides of the stem segments, and inoculate the stem segments into MS medium without hormones; Preferably, the 2% sodium hypochlorite solution is disinfected for 15min, the last sterile water is rinsed for 5 times, and the wedge-shaped block is 1cm long and 0.5cm thick.

[0008] Here, healthy and pest-free mother plants are selected, and strong growth stem segments with buds are cut.

[0009] The reason for selecting the morning on sunny days is that: the water content of the plant is moderate in the morning, avoiding rainwater carrying pathogenic bacteria, and the temperature is low, the microbial activity is weak, and the survival rate of the explant is improved.

[0010] Tween-20 is a non-ionic surfactant, which can make sodium hypochlorite wet and cover the surface and crevice of plant material evenly and sufficiently by reducing surface tension, so as to ensure that sodium hypochlorite can effectively contact with all potential microorganisms, thereby improving the disinfection efficiency and success rate and reducing pollution.

[0011] The inoculation into the MS medium without added hormones is because: if the disinfection scheme is preliminarily tried in the early stage, the brown dead and contaminated materials can be quickly distinguished, more observation time is provided, in addition, the transfer to the bud induction medium is generally in the state of germination, which is more efficient, if the bud induction medium is placed at the beginning, the nutrients in the medium will be consumed in two weeks, which affects the subsequent results.

[0012] S2: adventitious bud induction: the stem segments with buds which are disinfected and determined to survive are inoculated into the bud induction medium, the medium is: sucrose 10-40 g / L, MS medium 4-5 g / L, carrageenan 5-7 g / L, 6-BA 1.0 mg / L, IBA 0.3 mg / L, pH 5-6, and the culture conditions are: illumination 1000-2000 lx, temperature 20-26℃, illumination time 12-16 h / d; S3: bud proliferation: the obtained adventitious buds are transferred to the proliferation medium, the medium is: sucrose 10-40 g / L, MS medium 4-5 g / L, carrageenan 5-7 g / L, 6-BA 0.5 mg / L, IBA 0.3 mg / L, pH 5-6, and the culture conditions are: illumination 1000-2000 lx, temperature 20-26℃, illumination time 12-16 h / d; 6-benzyladenine (6-BA) is a cytokinin, which can promote cell division, elongation and differentiation, and has high stability and low cost; indole-3-butyric acid (IBA) is an auxin, which mainly regulates cell division, root differentiation and callus formation.

[0013] S4: strong seedling and rooting: the obtained cluster seedlings or single seedlings are transferred to the rooting medium, the medium is: sucrose 10-40 g / L, MS medium 4-5 g / L, carrageenan 5-7 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5-6, and the culture conditions are: illumination 1000-2000 lx, temperature 20-26℃, illumination time 12-16 h / d, to obtain the propagation system of Philodendron erubescens.

[0014] IBA (indole-3-butyric acid) can promote root elongation, and NAA (naphthalene acetic acid) can promote root thickening, and the combination of the two can make the roots grow fast and strong.

[0015] Further, the length of the stem section with buds in step S1 is 0.8-2 cm, and the plant is irrigated 3 days in advance.

[0016] Preferably, the length of the stem section with buds is 1 cm.

[0017] Selecting a proper length of the stem section can ensure sufficient bud points and avoid excessive incision to cause nutrient loss.

[0018] By irrigating in advance, the water content of the explant can be reduced, and the activity of endophytic bacteria can be reduced; carbendazim is a foliar fungicide, which can inhibit the germination of fungal spores, and reduce the pollution risk from the source.

[0019] Further, in step S2, the culture medium is: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.3 mg / L, pH 5.8, and the culture condition is: illumination 1500 lx, temperature 25℃, illumination time 14 h / d.

[0020] Further, in step S3, sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5 mg / L, IBA 0.3 mg / L, pH 5.8, and the culture condition is: illumination 1500 lx, temperature 25℃, illumination time 14 h / d.

[0021] Further, in step S4, sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8, and the culture condition is: illumination 1500 lx, temperature 25℃, illumination time 14 h / d.

[0022] Compared with the prior art, the present application has the following beneficial effects: (1) In the present application, Tween-20 can enhance the permeability of sodium hypochlorite, which is a non-ionic surfactant, can reduce the surface tension of sodium hypochlorite, so that sodium hypochlorite can more evenly and fully wet and cover the surface of plant material, especially the gap of stem section, to ensure that sodium hypochlorite can effectively contact all potential microorganisms, improve the disinfection efficiency and the survival rate of explants. Sodium hypochlorite (NaClO) destroys the cell membrane and protein structure of microorganisms by releasing active chlorine, and 15 min of treatment time can effectively kill bacteria and avoid oxidative damage to explants due to too long time. In addition, pre-water control and carbendazim spraying on the leaf surface are used for pretreatment of the plant, which can reduce the contamination of endophytic bacteria in the explants, pre-water control can reduce the water content of the plant and the activity of endophytic bacteria in the explants, and carbendazim is a broad-spectrum fungicide, which can inhibit the growth of fungi on the surface of the explants, further reducing the risk of explant contamination. Therefore, the method of Tween-20+2% NaClO disinfection treatment for 15 min is adopted in the present application, so that the survival rate of the explants reaches 73.3%, the browning rate is reduced to 11.1%, and the contamination rate is reduced to 15.6%, and it is low in toxicity and residue.

[0023] (2) In the present application, 6-BA and IBA synergistically regulate the cell division and differentiation of the Florida Philodendron stem section with buds, 6-benzyladenine (6-BA) as a cell division factor, activates the cell division of the bud primordium of the stem section with buds and the germination of the bud point. Indole-3-butyric acid (IBA) as an auxin, induces the formation of callus of the stem section with buds, and regulates the differentiation of bud primordium and the polar growth of bud, and synergistically improves the bud mass with 6-BA, so that the bud is thick and has no deformity. In the bud proliferation, the concentration of 6-BA is reduced to 0.5 mg / L, because the plant itself can produce hormones to maintain the subsequent subculture. If 1.0 mg / L 6-BA is continuously used, the number of buds will increase, but the buds are prone to be transparent, and it is difficult to grow into larger buds in subsequent subculture. The MS basic medium provides continuous nutrient supply of nitrogen and carbon source to meet the needs of rapid growth of buds. Therefore, the present application preferably uses the bud induction medium combination of 6-BA 1.0 mg / L+IBA 0.3 mg / L, and the bud proliferation medium combination of 6-BA 0.5 mg / L+IBA 0.3 mg / L, and the bud induction rate is as high as 94.4%, and the bud proliferation coefficient is 4.1.

[0024] (3) In the present application, IBA and NAA synergistically promote the root development of the Florida Philodendron stem section with buds, IBA (indole-3-butyric acid) promotes the formation of root primordium and the elongation of root system, and increases the number of root system. NAA (naphthalene acetic acid) induces root vascular bundle differentiation, thickens the root system, and improves the stress resistance of plant transplantation. The combination of the two can overcome the problem of weak or insufficient branching of plant root caused by single auxin, and achieve a plant rooting rate of 100% and a thick and strong root system.

[0025] (4) In the application, Tween-20 as a surfactant can make sodium hypochlorite enter the explant better, so that disinfection is more sufficient, and sodium hypochlorite is decomposed into non-toxic NaCl and H2O after cleaning, without heavy metal residue, meeting the requirements of green agriculture. Through optimization of disinfectant combination, hormone ratio and culture conditions, environmental safety is considered while high-efficiency propagation is realized, which provides reliable technical support for commercial production of Florida Philodendron.

[0026] (5) In the application, sodium hypochlorite is used to replace mercury, which is a high-efficiency and low-toxicity disinfection scheme, and the combination of 6-BA and IBA significantly improves the efficiency of bud induction and proliferation, and the whole process from disinfection to rooting is optimized, and a complete rapid propagation system is established. Experimental data show that 2% sodium hypochlorite and disinfection for 15 min, the survival rate is 73.3%, the pollution rate is 15.6%, and the brown death rate is 11.1%. Bud induction: 6-BA 1.0 mg / L + IBA 0.3 mg / L, bud germination rate 94.4%; bud proliferation: 6-BA 0.5 mg / L + IBA 0.3 mg / L, proliferation coefficient 4.1; rooting: IBA 0.2 mg / L + NAA 0.2 mg / L, rooting rate 100%.

[0027] It should be understood that the above general description and the following detailed description are only exemplary and explanatory, and cannot limit the application. DETAILED DESCRIPTION

[0028] The preferred embodiments of the application will be described in more detail below. Although the preferred embodiments of the application are shown, it should be understood that the application can be implemented in various forms and should not be limited by the embodiments described herein. On the contrary, these embodiments are provided to make the application more thorough and complete, and to fully convey the scope of the application to those skilled in the art.

[0029] The terms used in the present application are only for the purpose of describing specific embodiments, and are not intended to limit the application. The singular forms "a", "said" and "the" used in the present application and the appended claims are also intended to include the plural forms, unless the context clearly indicates otherwise. It should also be understood that the term "and / or" used herein means and includes any or all possible combinations of one or more associated listed items.

[0030] It should be understood that, although the terms "first", "second", "third", etc. can be used herein to describe various information, the information should not be limited to these terms. These terms are only used to distinguish the same type of information from each other. For example, the first information can also be referred to as the second information, and similarly, the second information can also be referred to as the first information without departing from the scope of the present application. Therefore, the features defined as "first", "second" can explicitly or implicitly include one or more of the features. In the description of the present application, the meaning of "a plurality of" is two or more, unless otherwise specifically limited.

[0031] Embodiment 1 The present embodiment discloses a method for establishing a rapid propagation system of Philodendron scandum, comprising the following steps: S1: treatment and disinfection of explants Irrigate the plants 3 days in advance, keep the potted soil dry, and spray the leaves with carbendazim; Sample in the morning on a sunny day, select healthy, pest-free mother plants, and cut the strong growth stem segments with buds to a length of 1 cm after preliminary pruning; Wash the surface dirt with laundry detergent and rinse thoroughly under running water; After ultraviolet disinfection for 30 minutes and sterile air ventilation for 20 minutes in the clean bench, the subsequent operation is performed. First, immerse the stem segments with buds in 75% alcohol for 30 seconds, constantly shake during the process to ensure that the disinfectant and the material are in full contact, then wash with sterile water for 3 times, then immerse the stem segments with buds in 2% sodium hypochlorite with 1 drop of Tween-20 for 15 minutes of disinfection treatment, after disinfection, wash with sterile water for 5 times until no foam appears on the surface of the stem segments with buds. Place the disinfected stem segments with buds on sterile filter paper to absorb the water, use sterilized instruments to cut into wedge-shaped blocks with bud points as the center, about 1 cm long and 0.5 cm thick, and make fresh wounds on three sides of the stem segments, and inoculate into MS medium without adding hormones.

[0032] S2: induction of adventitious buds Inoculate the disinfected and confirmed surviving stem segments with buds into bud induction medium; Medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.3 mg / L, pH 5.8; Cultivation conditions: light intensity 1500lx, temperature 25℃, light time 14h / d.

[0033] S3: bud proliferation Transfer the obtained adventitious buds into the proliferation medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5 mg / L, IBA 0.3 mg / L, pH 5.8; Culture conditions: illumination 1500 lx, temperature 25℃, illumination time 14 h / d.

[0034] S4: strong seedling and rooting The obtained cluster seedlings or single seedlings are transferred into a strong seedling and rooting culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8; Culture conditions: illumination 1500 lx, temperature 25℃, illumination time 14 h / d.

[0035] Example 2 The embodiment discloses a method for establishing a rapid propagation system of Agave cantula, comprising the following steps: S1: treatment and disinfection of explants The plants are irrigated 3 days in advance, the potting soil is kept dry, and the leaves are sprayed with carbendazim; The sample is taken in the morning on a sunny day, and a healthy mother plant without pests and diseases is selected, and a stem segment with strong growth force is cut and preliminarily trimmed to a length of 0.8 cm; The surface dirt is cleaned with washing powder and washed clean under running water; After ultraviolet disinfection for 30 min and sterile air ventilation for 20 min in the clean bench, subsequent operations are performed. First, the stem segment with buds is soaked in 75% alcohol for 30 s, and the disinfectant is constantly shaken to make the disinfectant fully contact with the material, and the stem segment with buds is washed with sterile water for 3 times, and then the stem segment with buds is soaked in 2% sodium hypochlorite added with 1 drop of Tween-20 for 15 min of disinfection treatment, and after disinfection, the stem segment with buds is washed with sterile water for 4 times until no foam appears on the surface of the stem segment with buds. The disinfected stem segment with buds is placed on sterile filter paper to absorb water, and the stem segment with buds is cut into a wedge-shaped block with a bud point as the center, about 0.5 cm long and about 0.3 cm thick, and fresh wounds are made on three sides of the stem segment, and inoculated into MS medium without adding hormones.

[0036] S2: induction of adventitious buds The disinfected and determined survival stem segment with buds is inoculated into a bud induction culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.1 mg / L, pH 5.8; Culture condition: light 1000lx, temperature 25℃, light time 14h / d.

[0037] S3: Bud proliferation The obtained adventitious buds were transferred into the proliferation culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.1 mg / L, pH 5.8; Culture condition: light 1000lx, temperature 25℃, light time 14h / d.

[0038] S4: Strong seedling and rooting The obtained cluster seedlings or single seedlings were transferred into the strong seedling and rooting culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8; Culture condition: light 1000lx, temperature 25℃, light time 14h / d.

[0039] Example 3 The embodiment discloses a method for establishing a rapid propagation system of Agave angustifolia Haw. S1: Treatment and disinfection of explants The plants were irrigated 3 days in advance, the potting soil was kept dry, and the leaves were sprayed with carbendazim; The sample was taken in the morning on a sunny day, and a healthy mother plant without pests and diseases was selected, and a stem segment with strong growth force was cut and preliminarily trimmed to a length of 1.2 cm; The surface dirt was cleaned with washing powder and washed clean under running water; After ultraviolet disinfection for 30 min and sterile air ventilation for 20 min in the clean bench, subsequent operations were performed. First, the stem segment with buds was soaked in 75% alcohol for 30 s, and the disinfectant was constantly shaken to make full contact with the material, and the stem segment with buds was washed with sterile water for 3 times, and then the stem segment with buds was soaked in 2% sodium hypochlorite added with 1 drop of Tween-20 for 15 min of disinfection treatment, and after disinfection, the stem segment with buds was washed with sterile water for 6 times until no foam appeared on the surface of the stem segment with buds. The disinfected stem segment with buds was placed on sterile filter paper to absorb water, and the stem segment with buds was cut into a wedge-shaped block with a bud point as the center, about 1.5 cm long and about 0.8 cm thick, and fresh wounds were made on three sides of the stem segment, and inoculated into the MS culture medium without adding hormones.

[0040] S2: Inducing adventitious buds The disinfected and determined survival stem segment with buds was inoculated into the bud induction culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.3 mg / L, pH 5.8; Culture condition: illumination 2000lx, temperature 25℃, illumination time 14h / d.

[0041] S3: bud proliferation The obtained adventitious buds are transferred into the proliferation culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5 mg / L, IBA 0.3 mg / L, pH 5.8; Culture condition: illumination 2000lx, temperature 25℃, illumination time 14h / d.

[0042] S4: strong seedling and rooting The obtained cluster seedlings or single seedlings are transferred into the strong seedling and rooting culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8; Culture condition: illumination 2000lx, temperature 25℃, illumination time 14h / d.

[0043] Effect verification: In order to further illustrate the effect of the present application, the following experiments and comparison are carried out: Experimental example 1 The embodiment discloses a method for establishing a fast propagation system of Agave angustifolia Haw. S1: treatment and disinfection of explants The plants are irrigated and watered 3 days in advance, the potted soil is kept dry, and the leaves are sprayed with carbendazim; The sample is taken in the morning on a sunny day, the healthy and pest-free mother plants are selected, the stem segments with strong growth force and buds are cut, and the length is preliminarily trimmed to 1cm; The surface dirt is cleaned with washing powder and washed clean under running water; After ultraviolet sterilization for 30 min and sterile air ventilation for 20 min in a clean bench, subsequent operations are performed. First, the stem segments with buds are soaked in 75% alcohol for 30 s, with constant shaking to ensure that the disinfectant fully contacts the material. The stem segments with buds are washed with sterile water for 3 times, and then are soaked in 2% sodium hypochlorite with 1 drop of Tween-20 for 15 min of disinfection treatment. After disinfection, the stem segments with buds are washed with sterile water for 5 times until no foam appears on the surface of the stem segments with buds. The disinfected stem segments with buds are placed on sterile filter paper to dry the water, and are cut into wedge-shaped blocks with a bud point as the center, about 1 cm long and about 0.5 cm thick, using sterilized instruments. Fresh wounds are made on three sides of the stem segments, and are inoculated into MS medium without added hormones.

[0044] S2: Inducing the generation of adventitious buds The disinfected and determined survival stem segments with buds are inoculated into the bud induction medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.3 mg / L, pH 5.8; Culture conditions: light intensity 1500 lx, temperature 25°C, light time 14 h / d.

[0045] S3: Bud proliferation The obtained adventitious buds are transferred into the proliferation medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5 mg / L, IBA 0.3 mg / L, pH 5.8; Culture conditions: light intensity 1500 lx, temperature 25°C, light time 14 h / d.

[0046] S4: Strong seedling and rooting The obtained clump seedlings or single seedlings are transferred into the strong seedling and rooting medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8; Culture conditions: light intensity 1500 lx, temperature 25°C, light time 14 h / d.

[0047] Experimental Example 2 The embodiment discloses a method for establishing a rapid propagation system of Philodendron erubescens, comprising the following steps: S1: Disinfection of explants The plants are irrigated 3 days in advance, the potting soil is kept dry, and the leaves are sprayed with carbendazim; Sample in the morning of sunny day, select healthy, no disease and pest mother plant, cut the stem section with strong growth force, and cut to 0.8cm in length; Use washing powder to clean the surface dirt, and rinse under running water; After ultraviolet disinfection for 30 min and sterile air ventilation for 20 min in the super-clean bench, subsequent operations are carried out. Firstly, the stem section with bud is soaked in 75% alcohol for 30 s, and the disinfectant is fully contacted with the material by constantly shaking, and the stem section with bud is washed with sterile water for 3 times, and then the stem section with bud is soaked in 2% sodium hypochlorite added with 1 drop of Tween-20 for 10 min of disinfection treatment, and after disinfection, the stem section with bud is washed with sterile water for 4 times until no foam appears on the surface of the stem section with bud. The stem section with bud after disinfection is placed on sterile filter paper to absorb water, and the stem section with bud is cut into a wedge-shaped block with a bud point as the center, about 0.5 cm long and about 0.3 cm thick, and fresh wounds are made on three sides of the stem section, and inoculated into MS medium without added hormones.

[0048] S2: Inducing the generation of adventitious buds The stem section with bud after disinfection and determined survival is inoculated into the bud induction medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.1 mg / L, pH 5.8; Culture condition: light 1000lx, temperature 25℃, light time 14h / d.

[0049] S3: Bud proliferation The obtained adventitious buds are transferred into the proliferation medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.1 mg / L, pH 5.8; Culture condition: light 1000lx, temperature 25℃, light time 14h / d.

[0050] S4: Strong seedling and rooting The obtained cluster seedlings or single seedlings are transferred into the strong seedling and rooting medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8; Culture condition: light 1000lx, temperature 25℃, light time 14h / d.

[0051] Experimental example 3 The embodiment discloses a method for establishing a rapid propagation system of Philodendron erubescens, comprising the following steps: S1: Explant treatment and disinfection Irrigate the plants 3 days in advance, keep the potting soil dry, and spray the leaves with carbendazim; Sample in the morning on a sunny day, select healthy, disease-free mother plants, and cut the strong growth stem segments with buds to 1.2 cm in length after preliminary pruning; Wash the surface dirt with laundry detergent and rinse thoroughly under running water; After 30 minutes of ultraviolet disinfection and 20 minutes of sterile air ventilation in the clean bench, the subsequent operation is performed. First, immerse the stem segments with buds in 75% alcohol for 30 seconds, constantly shake during the process to ensure that the disinfectant fully contacts the material, then wash with sterile water for 3 times, then immerse the stem segments with buds in 2% sodium hypochlorite with 1 drop of Tween-20 for 15 minutes of disinfection treatment, after disinfection, wash with sterile water for 6 times until no foam appears on the surface of the stem segments with buds. Place the disinfected stem segments with buds on sterile filter paper to absorb the water, use sterilized instruments to cut into wedge-shaped blocks with bud points as the center, about 1.5 cm long and 0.8 cm thick, and make fresh wounds on three sides of the stem segments, and inoculate into MS medium without added hormones.

[0052] S2: Inducing adventitious buds Inoculate the disinfected and confirmed surviving stem segments with buds into bud induction medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.3 mg / L, pH 5.8; Culture conditions: light intensity 2000 lx, temperature 25°C, light time 14 h / d.

[0053] S3: Bud proliferation Transfer the obtained adventitious buds into the proliferation medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5 mg / L, IBA 0.3 mg / L, pH 5.8; Culture conditions: light intensity 2000 lx, temperature 25°C, light time 14 h / d.

[0054] S4: Strong seedlings and rooting Transfer the obtained clumps or single seedlings into the strong seedling and rooting medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8; Culture conditions: light intensity 2000 lx, temperature 25°C, light time 14 h / d.

[0055] Comparative Example 1 The embodiment discloses a method for establishing a rapid propagation system of Philodendron scandum, and comprises the following steps: S1: treatment and disinfection of explants Irrigate the plants 3 days in advance, keep the potted soil dry, and spray the leaves with carbendazim; Sample in the morning on a sunny day, select healthy and pest-free mother plants, and cut the stem segments with strong growth force to a length of 1 cm after preliminary pruning; Wash the surface dirt with laundry detergent and rinse thoroughly under running water; After ultraviolet disinfection for 30 minutes and sterile air ventilation for 20 minutes in the clean bench, the subsequent operation is performed. First, immerse the stem segments with buds in 75% alcohol for 30 seconds, constantly shake during the process to ensure that the disinfectant and the material are in full contact, wash with sterile water for 3 times, then immerse the stem segments with buds in 1% sodium hypochlorite with 1 drop of Tween-20 for 15 minutes of disinfection treatment, after disinfection, wash with sterile water for 5 times until no foam appears on the surface of the stem segments with buds. Place the disinfected stem segments with buds on sterile filter paper to absorb water, use sterilized instruments to cut into wedge-shaped blocks with bud points as the center, about 1 cm long and 0.5 cm thick, and make fresh wounds on three sides of the stem segments, and inoculate into MS medium without adding hormones.

[0056] S2: induction of adventitious buds Inoculate the disinfected and determined survival stem segments with buds into the bud induction medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5 mg / L, IBA 0.3 mg / L, pH 5.8; Culture conditions: illumination 1500lx, temperature 25℃, illumination time 14h / d.

[0057] S3: bud proliferation Transfer the obtained adventitious buds into the proliferation medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.3 mg / L, pH 5.8; Culture conditions: illumination 1500lx, temperature 25℃, illumination time 14h / d.

[0058] S4: strong seedling and rooting Transfer the obtained cluster seedlings or single seedlings into the strong seedling and rooting medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8; Culture conditions: illumination 1500lx, temperature 25℃, illumination time 14h / d.

[0059] Comparative example 2 The embodiment discloses a method for establishing a rapid propagation system of Florida Philodendron, comprising the following steps: S1: treatment and disinfection of explants Irrigate the plants 3 days in advance, keep the potted soil dry, and spray the leaves with carbendazim; Sample in the morning on a sunny day, select healthy and pest-free mother plants, and cut the stem segments with strong growth force, and initially trim to a length of 1cm; Wash the surface dirt with washing powder and rinse under running water until clean; After ultraviolet disinfection for 30 minutes and sterile air ventilation for 20 minutes in the clean bench, subsequent operations are performed. First, immerse the stem segments with buds in 75% alcohol for 30 seconds, constantly shake during the process to make the disinfectant fully contact with the material, wash with sterile water for 3 times, then immerse the stem segments with buds in 1% sodium hypochlorite with 1 drop of Tween-20 for 10 minutes of disinfection treatment, after disinfection, wash with sterile water for 5 times until no foam appears on the surface of the stem segments with buds. Place the disinfected stem segments with buds on sterile filter paper to absorb water, use sterilized instruments to cut into wedge-shaped blocks with a bud point as the center, about 1cm long and about 0.5cm thick, make fresh wounds on three sides of the stem segments, and inoculate into MS medium without adding hormones.

[0060] S2: induction of adventitious buds Inoculate the disinfected and determined survival stem segments with buds into the bud induction medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5mg / L, IBA 0.1mg / L, pH 5.8; Culture conditions: illumination 1500lx, temperature 25℃, illumination time 14h / d.

[0061] S3: bud proliferation Transfer the obtained adventitious buds into the proliferation medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0mg / L, IBA 0.5mg / L, pH 5.8; Culture conditions: illumination 1500lx, temperature 25℃, illumination time 14h / d.

[0062] S4: Strengthening and rooting of seedlings The obtained cluster seedlings or single seedlings are transferred into a medium for strengthening and rooting; Medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8; Culture conditions: illumination 1500 lx, temperature 25°C, illumination time 14 h / d.

[0063] Comparative Example 3 The embodiment discloses a method for establishing a rapid propagation system of Agave angustifolia Haw., comprising the following steps: S1: Treatment and disinfection of explants The plants are irrigated 3 days in advance, the potting soil is kept dry, and the leaves are sprayed with carbendazim; The sample is taken in the morning on a sunny day, and a healthy mother plant without pests and diseases is selected, and a stem segment with strong growth force is cut and trimmed to a length of 1 cm; The surface dirt is cleaned with laundry detergent and washed clean under running water; After ultraviolet disinfection for 30 min and sterile air ventilation for 20 min in a clean bench, subsequent operations are performed. First, the stem segment with buds is soaked in 75% alcohol for 30 s, and the disinfectant is constantly shaken to make full contact with the material. The stem segment with buds is washed with sterile water for 3 times, and then the stem segment with buds is soaked in 1% sodium hypochlorite with 1 drop of Tween-20 for 20 min of disinfection treatment. After disinfection, the stem segment with buds is washed with sterile water for 5 times until no foam appears on the surface of the stem segment with buds. The disinfected stem segment with buds is placed on sterile filter paper to absorb water, and the stem segment with buds is cut into a wedge-shaped block with a bud point as the center, about 1 cm long and about 0.5 cm thick, and fresh wounds are made on three sides of the stem segment, and inoculated into MS medium without adding hormones.

[0064] S2: Inducing adventitious buds The disinfected and determined survival stem segment with buds is inoculated into a bud induction medium; Medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5 mg / L, IBA 0.5 mg / L, pH 5.8; Culture conditions: illumination 1500 lx, temperature 25°C, illumination time 14 h / d.

[0065] S3: Bud proliferation The obtained adventitious buds are transferred into a proliferation medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5 mg / L, IBA 0.1 mg / L, pH 5.8; Culture conditions: illumination 1500lx, temperature 25℃, illumination time 14h / d.

[0066] S4: strong seedling and rooting The obtained cluster seedlings or single seedlings are transferred into a strong seedling and rooting culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8; Culture conditions: illumination 1500lx, temperature 25℃, illumination time 14h / d.

[0067] Comparative example 4 The embodiment discloses a method for establishing a rapid propagation system of Agave cantula, and comprises the following steps: S1: treatment and disinfection of explants The plants are irrigated 3 days in advance, the potting soil is kept dry, and carbendazim is sprayed on the leaves; The sample is taken in the morning on a sunny day, and a healthy mother plant without pests and diseases is selected, and a stem segment with strong growth force and buds is cut and trimmed to a length of 1cm; The surface dirt is cleaned with washing powder and washed clean under running water; After ultraviolet disinfection for 30 minutes and sterile air ventilation for 20 minutes in the clean bench, subsequent operations are performed. First, the stem segment with buds is soaked in 75% alcohol for 30 seconds, and the disinfectant is constantly shaken to make full contact with the material, and the stem segment with buds is washed with sterile water for 3 times, and then the stem segment with buds is soaked in 3% sodium hypochlorite with 1 drop of Tween-20 for 15 minutes of disinfection treatment, and after disinfection, the stem segment with buds is washed with sterile water for 5 times until no foam appears on the surface of the stem segment with buds. The disinfected stem segment with buds is placed on sterile filter paper to absorb water, and the stem segment with buds is cut into a wedge-shaped block with a bud point as the center, about 1 cm long and about 0.5 cm thick, and fresh wounds are made on three sides of the stem segment, and inoculated into MS medium without adding hormones.

[0068] S2: induction of adventitious buds The disinfected and determined survival stem segment with buds is inoculated into a bud induction culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.5 mg / L, IBA 0.3 mg / L, pH 5.8; Culture conditions: illumination 1500lx, temperature 25℃, illumination time 14h / d.

[0069] S3: Bud multiplication The obtained adventitious buds are transferred into a multiplication culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5 mg / L, IBA 0.5 mg / L, pH 5.8; Culture conditions: illumination 1500 lx, temperature 25°C, illumination time 14 h / d.

[0070] S4: Strong seedling and rooting The obtained cluster seedlings or single seedlings are transferred into a strong seedling and rooting culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8; Culture conditions: illumination 1500 lx, temperature 25°C, illumination time 14 h / d Comparative Example 5 The embodiment discloses a method for establishing a rapid propagation system of Agave cantula, comprising the following steps: S1: Treatment and disinfection of explants The plants are irrigated 3 days in advance, the potting soil is kept dry, and the leaves are sprayed with carbendazim; The sampling is performed in the morning on a sunny day, and a healthy mother plant free of pests and diseases is selected, and a stem segment with strong growth force is cut and preliminarily trimmed to a length of 1 cm; The surface dirt is cleaned with washing powder and washed clean under running water; After ultraviolet disinfection for 30 min and sterile air ventilation for 20 min in a clean bench, subsequent operations are performed. First, the stem segment with buds is soaked in 75% alcohol for 30 s, and the disinfectant is constantly shaken to make full contact with the material, and the stem segment with buds is washed with sterile water for 3 times, and then the stem segment with buds is soaked in 3% sodium hypochlorite added with 1 drop of Tween-20 for 10 min of disinfection treatment, and after disinfection, the stem segment with buds is washed with sterile water for 5 times until no foam appears on the surface of the stem segment with buds. The disinfected stem segment with buds is placed on sterile filter paper to absorb water, and a sterilized instrument is used to cut a wedge-shaped piece with a bud point as the center, about 1 cm long and about 0.5 cm thick, and fresh wounds are made on three sides of the stem segment, and inoculated into MS medium without adding hormones.

[0071] S2: Inducing generation of adventitious buds The disinfected and determined survival stem segment with buds is inoculated into a bud induction culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.3 mg / L, pH 5.8; Culture conditions: illumination 1500 lx, temperature 25℃, illumination time 14 h / d.

[0072] S3: bud proliferation The obtained adventitious buds are transferred into the proliferation culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5 mg / L, IBA 0.3 mg / L, pH 5.8; Culture conditions: illumination 1500 lx, temperature 25℃, illumination time 14 h / d.

[0073] S4: seedling strengthening and rooting The obtained cluster seedlings or single seedlings are transferred into the seedling strengthening and rooting culture medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8; Culture conditions: illumination 1500 lx, temperature 25℃, illumination time 14 h / d Comparative example 6 The embodiment discloses a method for establishing a rapid propagation system of Philodendron erubescens, and comprises the following steps: S1: treatment and disinfection of explants The plants are irrigated 3 days in advance, the potted soil is kept dry, and the leaves are sprayed with carbendazim; The sampling is performed in the morning on a sunny day, and a healthy mother plant free of pests and diseases is selected, and a stem segment with strong growth force and buds is cut and trimmed to a length of 1 cm; The surface dirt is cleaned with washing powder and washed clean under running water; After ultraviolet disinfection for 30 min and sterile air ventilation for 20 min in the clean bench, subsequent operations are performed. First, the stem segment with buds is soaked in 75% alcohol for 30 s, and the disinfectant is constantly shaken to make full contact with the material, and the stem segment with buds is washed with sterile water for 3 times, and then the stem segment with buds is soaked in 3% sodium hypochlorite added with 1 drop of Tween-20 for 20 min of disinfection treatment, and after disinfection, the stem segment with buds is washed with sterile water for 5 times until no foam appears on the surface of the stem segment with buds. The disinfected stem segment with buds is placed on sterile filter paper to absorb water, and the stem segment with buds is cut into a wedge-shaped block with a bud point as the center, about 1 cm long and about 0.5 cm thick, and fresh wounds are made on three sides of the stem segment, and inoculated into the MS culture medium without adding hormones.

[0074] S2: Inducing adventitious buds The sterilized and determined survival stem segments with buds were inoculated into bud induction medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.3 mg / L, pH 5.8; Culture conditions: illumination 1500 lx, temperature 25℃, illumination time 14 h / d.

[0075] S3: Bud proliferation The obtained adventitious buds were transferred into the proliferation medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5 mg / L, IBA 0.3 mg / L, pH 5.8; Culture conditions: illumination 1500 lx, temperature 25℃, illumination time 14 h / d.

[0076] S4: Strong seedling and rooting The obtained clump seedlings or single seedlings were transferred into the strong seedling and rooting medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8; Culture conditions: illumination 1500 lx, temperature 25℃, illumination time 14 h / d.

[0077] Comparative example 7 The embodiment discloses a method for establishing a rapid propagation system of Florida Philodendron, comprising the following steps: S1: Treatment and disinfection of explants The plants were irrigated and watered 3 days in advance, the potted soil was kept dry, and the leaves were sprayed with carbendazim; The samples were taken in the morning on sunny days, and healthy and pest-free mother plants were selected, and stem segments with strong growth force and buds were cut and trimmed to a length of 1 cm; The surface dirt was cleaned with laundry detergent and washed clean under running water; After ultraviolet sterilization for 30 min and sterile air ventilation for 20 min in a clean bench, subsequent operations were carried out. First, the stem segments with buds were soaked in 75% alcohol for 30 s, with constant shaking to ensure that the disinfectant fully contacted the material. The stem segments with buds were then washed with sterile water for 3 times, and then soaked in 2% sodium hypochlorite solution containing 1 drop of Tween-20 for 20 min of disinfection treatment. After disinfection, the stem segments with buds were washed with sterile water for 5 times until no foam appeared on the surface of the stem segments with buds. The disinfected stem segments with buds were placed on sterile filter paper to dry, and then cut into wedge-shaped blocks with a bud point as the center, about 1 cm long and about 0.5 cm thick, using sterilized instruments. Fresh wounds were made on three sides of the stem segments, and then inoculated into MS medium without added hormones.

[0078] S2: Inducing the formation of adventitious buds The disinfected and determined survival stem segments with buds were inoculated into bud induction medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.5 mg / L, pH 5.8; Culture conditions: light intensity 1500 lx, temperature 25℃, light time 14 h / d.

[0079] S3: Bud proliferation The obtained adventitious buds were transferred into the proliferation medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5 mg / L, IBA 0.3 mg / L, pH 5.8; Culture conditions: light intensity 1500 lx, temperature 25℃, light time 14 h / d.

[0080] S4: Strengthening seedlings and rooting The obtained clump seedlings or single seedlings were transferred into the strengthening seedling and rooting medium; Culture medium formula: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8; Culture conditions: light intensity 1500 lx, temperature 25℃, light time 14 h / d Effects of different sodium hypochlorite concentrations and disinfection times on the contamination rate, brown mortality rate and survival rate of explants The establishment of the rapid propagation system of Florida Philodendron in Experimental Examples 1-2 and Comparative Examples 1-7 was determined. In Experimental Examples 1-2, 2% sodium hypochlorite was used for 10-15 min of disinfection treatment. In Comparative Examples 1-7, 1% and 3% sodium hypochlorite were used for 10-20 min of disinfection treatment, respectively.

[0081] Experimental method: the stem segments with buds were soaked in 75% alcohol for 30s, with constant shaking to ensure that the disinfectant fully contacted the material, and then washed with sterile water for 3 times. Then the stem segments with buds were soaked in 1-3% sodium hypochlorite with 1 drop of Tween-20 for 10-20 min disinfection treatment, and washed with sterile water for 5 times after disinfection until no foam appeared on the surface of the stem segments with buds. The disinfected stem segments with buds were placed on sterile filter paper to dry the water, and then cut into wedge-shaped blocks with a bud point as the center, about 1 cm long and 0.5 cm thick using sterilized instruments, and fresh wounds were made on three sides of the stem segments, and then inoculated into MS medium without added hormones for culture, and the contamination rate, browning rate and survival rate of the stem segments with buds were observed and recorded.

[0082] Experimental results: in Table 1, the contamination rate was the lowest when treated with 2% sodium hypochlorite for 15 min, which was 15.6%, significantly lower than that treated with 1% and 3% sodium hypochlorite, which were 28.9% and 7.8% respectively. The contamination rate was 0% when treated with 3% sodium hypochlorite for 20 min, but the browning rate was very high, which was 72.2%, indicating that high concentration and long time disinfection could completely sterilize the explants, but seriously damaged the explants.

[0083] The browning rate was the lowest when treated with 2% sodium hypochlorite for 15 min, which was 11.1%, much lower than that treated with 3% sodium hypochlorite for 15 min, which was 43.3%. The browning rate was as high as 46.7% when treated with 1% sodium hypochlorite for 20 min, indicating that low concentration and long time disinfection still easily led to browning.

[0084] The survival rate was the highest when treated with 2% sodium hypochlorite for 15 min, which was 73.3%, significantly better than other treatments, such as 36.7% when treated with 1% sodium hypochlorite for 15 min. Although 3% sodium hypochlorite could reduce the contamination rate, the survival rate decreased significantly due to the increase of browning rate.

[0085] Therefore, the optimal scheme was 2% sodium hypochlorite + 15 min disinfection, which considered low contamination rate of 15.6%, low browning rate of 11.1% and high survival rate of 73.3%.

[0086] Table 1 Results of stem segment disinfection and multiple comparison table NaClO concentration Time Contamination rate (%) Brown mortality rate (%) Survival rate (%) 1% 10 min 42.2±1.6 aA 26.7±2.7 bB 31.1±3.1 bB 1% 15 min 28.9±4.2 aA 34.4±5.7 bB 36.7±5.4 bB 1% 20 min 17.8±1.6 aA 46.7±2.7 bB 35.6±1.6 bB 2% 10 min 16.8±2.7 bB 22.2±4.2 bB 61.1±4.2 aA 2% 15 min 15.6±3.1 bB 11.1±1.6 bB 73.3±2.7 aA 2% 20 min 3.3±2.7 bB 40.0±2.7 bB 56.7±5.4 aA 3% 10 min 10.0±2.7 bB 44.4±5.7 aA 45.6±3.1 bB 3% 15 min 7.8±4.2 bB 43.3±4.7 aA 48.9±3.1 bB 3% 20 min 0.0±0.0 bB 72.2±4.2 aA 27.8±4.2 bB Note: the data in the table are mean ± standard error. Three replicates, each replicate is 30 explants. The data were tested by Duncan's new multiple range method (a, b = 0.05, A, B = 0.01), and different lowercase letters in the same column indicate significant difference at 0.05 level.

[0087] 2, Effect of different concentrations of 6-BA and IBA on bud induction The establishment of the fast propagation system of experimental examples 1-2 and comparative examples 1-7 Florida Philodendron was determined, wherein experimental examples 1-2 refer to bud induction of 6-BA 1.0 mg / L + IBA 0.1-0.3 mg / L combination, and comparative examples 1-7 refer to bud induction of 6-BA 0.5 mg / L + IBA 0.1-0.5 mg / L combination, bud induction of 6-BA 1.5 mg / L + IBA 0.1-0.5 mg / L combination, and bud induction of 6-BA 1.0 mg / L + IBA 0.5 mg / L combination.

[0088] Experimental method: the sterilized and determined survival stem segments with buds were inoculated in the bud induction medium, and the medium formula was sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5-1.5 mg / L, IBA 0.1-0.5 mg / L, pH 5.8; the culture conditions were illumination 1500 lx, temperature 25℃, and illumination time 14 h / d.

[0089] Experimental results: in table 2, the bud germination rate of 6-BA 1.0 mg / L + IBA 0.3 mg / L combination was the highest, which was 94.4%, which was significantly better than other combinations, such as 6-BA 0.5 mg / L + IBA 0.5 mg / L was only 66.7%, when the concentration of 6-BA was 1.5 mg / L, the bud germination rate decreased significantly, which was 50.0%, indicating that high concentration of cytokinin may inhibit bud differentiation.

[0090] The bud body of 6-BA 1.0 mg / L + IBA 0.3 mg / L grew well, while the bud body of 6-BA 1.5 mg / L + IBA 0.5 mg / L grew weakly and the germination rate was low, which was 32.2%, when the concentration of IBA was too high, which was 0.5 mg / L, it was easy to lead to the overgrowth of bud body and weak growth.

[0091] Therefore, 6-BA 1.0 mg / L + IBA 0.3 mg / L is the best bud induction hormone ratio, the bud germination rate is 94.4%, and the bud body is strong.

[0092] Table 2 bud induction results of different plant growth regulators and multiple comparison table 6-BA (mg / L) IBA (mg / L) Average number of sprouted explants per explant Average sprouting rate (%) Growth condition 0.5 0.1 17.3 57.8 eEF Slow sprouting, few sprouts, good growth 0.5 0.3 22.3 74.4 cCD Slow sprouting, few sprouts, strong growth 0.5 0.5 20.0 66.7 dDE Fast sprouting, many sprouts, weak growth 1.0 0.1 25.3 84.4 bB Fast sprouting, many sprouts, good growth 1.0 0.3 28.3 94.4 aA Fast sprouting, many sprouts, strong growth 1.0 0.5 23.0 76.7 cBC Fast sprouting, many sprouts, weak growth 1.5 0.1 21.3 71.1 cdCD Fast sprouting, many sprouts, weak growth 1.5 0.3 15.0 50.0 fF Slow sprouting, few sprouts, strong growth 1.5 0.5 9.7 32.2 gG Slow sprouting, few sprouts, weak growth Note: the data in the table is mean ± standard error. Three repetitions, each repetition is 30 explants. The data is tested by Duncan's new multiple range method (a, b=0.05, A, B=0.01), and different lowercase letters in the same column represent significant difference at 0.05 level.

[0093] 3, the effect of different 6-BA and IBA concentrations on bud multiplication The establishment of the fast propagation system of experimental examples 1~2 and comparative examples 1~4 Florida Philodendron was measured, wherein experimental examples 1~2 refer to the bud induction of 6-BA 0.5 mg / L + IBA 0.3 mg / L combination, the bud induction of 6-BA 1.0 mg / L + IBA 0.1 mg / L combination, and comparative examples 1~4 refer to the bud induction of 6-BA 0.5 + IBA 0.1 mg / L combination, the bud induction of 6-BA 0.5 + IBA 0.5 mg / L combination, and the bud induction of 6-BA 1.0 mg / L + IBA 0.3-0.5 mg / L combination.

[0094] Experimental method: the obtained adventitious buds were transferred into the proliferation medium, the medium formula was sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5-1.0 mg / L, IBA 0.1-0.5 mg / L, pH 5.8; the culture conditions were light 1500 lx, temperature 25℃, light time 14 h / d.

[0095] Experimental results: in table 3, the proliferation coefficient of 6-BA 0.5 mg / L + IBA 0.3 mg / L was the highest, which was 4.1, which was significantly better than other combinations, such as 6-BA 1.0 mg / L + IBA 0.3 mg / L, which was 3.1, when the combination was 6-BA 1.0 mg / L + IBA 0.5 mg / L, the proliferation coefficient was very low, which was 1.1, which indicated that high concentration of IBA inhibited bud proliferation.

[0096] The bud body of 6-BA 0.5 mg / L + IBA 0.3 mg / L was dense and the leaf color was dark green, while the bud body of 6-BA 1.0 mg / L + IBA 0.5 mg / L was yellow and grew poorly. Therefore, 6-BA 0.5 mg / L + IBA 0.3 mg / L was the best bud proliferation hormone ratio, the proliferation coefficient was 4.1, and the bud body was strong.

[0097] Table 3 bud proliferation results of different plant growth regulators and multiple comparison table 6-BA (mg / L) IBA (mg / L) Proliferation coefficient Growth condition 0.5 0.1 2.0±0.1 eD Few sprouts, slow growth, and light green leaves 0.5 0.3 4.1±0.0 aA Dense and strong sprouts, dark green leaves 0.5 0.5 2.9±0.1 dC Uniformly distributed and strong sprouts, slow growth, and light green leaves 1.0 0.1 3.5±0.0 bB Uniformly distributed and strong sprouts, moderate leaf color 1.0 0.3 3.1±0.1 cC Uniformly distributed sprouts, weak sprouts, and light green leaves 1.0 0.5 1.1±0.1 fE Few sprouts, and yellowish and withered leaves Note: the data in the table is mean ± standard error. Three repetitions, each repetition is 30 explants. The data is tested by Duncan's new multiple range method (a, b=0.05, A, B=0.01).

[0098] Having described various embodiments of the application, it is to be understood that the above description is meant to be illustrative only and not exhaustive. Modifications, changes and variations which are obvious to those of ordinary skill in the art are intended to be within the scope of the application. The foregoing description details certain embodiments of the application. It will be appreciated, however, that no matter how detailed the above appears in text, the application can be practiced in many ways, as will be understood by those skilled in the art. Therefore, the application should not be construed as limited to the embodiments described above. Rather, the scope of the application should be measured by the broadest interpretation of the claims that are included below.

Claims

1. A method for establishing a rapid propagation system of Aglaia flori dae, characterized by, Comprising the following steps: S1: explant treatment and disinfection: irrigate the plants in advance, keep the potted soil dry, spray the plant leaves with carbendazim, select healthy and disease-free mother plants in the morning on sunny days, cut the stem segments with strong growth force, wash the leaf surface of the stem segments with washing powder, then rinse with running water, and then perform the following treatment on the cleaned stem segments in the clean bench: soak in 75% alcohol for 30s, constantly shake during the process, then rinse with sterile water for 3 times, then soak the stem segments in 2% sodium hypochlorite solution containing 1 drop of Tween-20 for 15min, rinse with sterile water for 4-6 times after disinfection until no foam appears on the surface of the stem segments, place the disinfected stem segments on sterile filter paper to absorb water, cut the stem segments into wedge-shaped blocks with a bud point as the center, about 0.5-1.5cm long and 0.3-0.8cm thick using sterilized instruments, and make fresh wounds on three sides of the stem segments, and inoculate the stem segments into MS medium without hormones; S2: adventitious bud induction: inoculate the disinfected and determined survival stem segments into the bud induction medium, the medium is: sucrose 10-40 g / L, MS medium 4-5 g / L, carrageenan 5-7 g / L, 6-BA 1.0 mg / L, IBA 0.3 mg / L, pH 5-6, the culture conditions are: illumination 1000-2000lx, temperature 20-26℃, illumination time 12-16h / d; S3: bud proliferation: transfer the obtained adventitious buds to the proliferation medium, the medium is: sucrose 10-40 g / L, MS medium 4-5 g / L, carrageenan 5-7 g / L, 6-BA 0.5 mg / L, IBA 0.3 mg / L, pH 5-6, the culture conditions are: illumination 1000-2000lx, temperature 20-26℃, illumination time 12-16h / d; S4: strong seedling and rooting: transfer the obtained cluster seedlings or single seedlings to the rooting medium, the medium is: sucrose 10-40 g / L, MS medium 4-5 g / L, carrageenan 5-7 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5-6, the culture conditions are: illumination 1000-2000lx, temperature 20-26℃, illumination time 12-16h / d, and the Floridagreenery propagation system is obtained.

2. The method for establishing the Aglaia floridana rapid propagation system according to claim 1, characterized in that, The length of the stem segments in step S1 is 0.8-2cm, and the plants are irrigated 3d in advance.

3. The method for establishing the Aglaia rottleroides micropropagation system according to claim 1, wherein, In step S2, the medium is: sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 1.0 mg / L, IBA 0.3 mg / L, pH 5.8, and the culture conditions are: illumination 1500lx, temperature 25℃, illumination time 14h / d.

4. The method for establishing the Agapanthus africanus rapid propagation system according to claim 1, wherein, In step S3, sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, 6-BA 0.5 mg / L, IBA 0.3 mg / L, pH 5.8, the culture conditions are: illumination 1500 lx, temperature 25℃, illumination time 14 h / d.

5. The method for establishing the Aglaia rottleroides micropropagation system according to claim 1, wherein, In step S4, sucrose 30 g / L, MS medium 4.74 g / L, carrageenan 6 g / L, IBA 0.2 mg / L, NAA 0.2 mg / L, pH 5.8, the culture conditions are: illumination 1500 lx, temperature 25℃, illumination time 14 h / d.