Cuttage planting method for caulis spatholobi

Abstract

The invention provides a suberect spatholobus stem cutting planting method, and relates to the technical field of plant cutting seedling raising. The cuttage planting method comprises the steps that caulis spatholobi cuttage branches are disinfected and then soaked with rooting liquid prepared from stenotrophomonas maltophilia plant fermentation post-treatment liquid, indoleacetic acid, salicylic acid and rice milk, and then cuttage seedling raising is conducted. According to the method, the defects in the prior art are overcome, the fermentation conditions of the stenotrophomonas maltophilia are optimized, sterilization treatment is conducted at the same time, the sterilized post-treatment liquid is obtained and serves as a main functional component of the rooting liquid, early rooting of the caulis spatholobi cutting seedlings is effectively promoted, the overall survival rate is increased, and the cost of the caulis spatholobi cutting seedlings is reduced.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of plant cutting seedling, in particular to a cutting planting method of Spatholobus suberectus. BACKGROUND

[0002] Spatholobus suberectus is the dried stem of Spatholobus suberectus of the genus Millettia of the Leguminosae, which belongs to traditional Chinese medicinal materials in China. Spatholobus suberectus contains flavonoids, phenolic acids, terpenes and sterols and other active ingredients, and has a wide market. Due to the wide market demand, the wild Spatholobus suberectus resources cannot meet the market demand, so at present artificial planting Spatholobus suberectus is mainly used.

[0003] The propagation methods of Spatholobus suberectus mainly include seed propagation and cutting propagation. Since the overall seed setting rate of Spatholobus suberectus is low and the seed picking is difficult, the artificial propagation method commonly used at present is cutting propagation. However, the survival rate of the existing Spatholobus suberectus cutting propagation is not stable, and the survival rate fluctuates between 20% and 85%. The survival rate of cutting is mainly affected by the type and concentration of rooting agent.

[0004] The existing method for improving the survival rate of Spatholobus suberectus cutting is mainly to soak plant growth hormones to promote rooting of cutting. However, the overall survival rate is basically around 80%, which causes waste of a part of branches. In addition, Spatholobus suberectus generally roots after 10-15 days of cutting, and the early root growth efficiency is slow. Therefore, in order to reduce the cost of artificial seedling of Spatholobus suberectus, improving the survival rate of cutting seedling is an important research direction at present.

[0005] Stenotrophomonas maltophilia is a kind of gram-negative aerobic bacillus, which can secrete organic acids and other substances to convert insoluble phosphate in soil into available phosphorus for plants. In addition, to a certain extent, it can secrete growth regulators in soil to directly stimulate rooting. Therefore, it is added as a beneficial microbial agent in the growth process of some plants. However, the current research on stenotrophomonas maltophilia for promoting rooting is mainly in the field of conventional seed propagation. There is a significant difference in mechanism between plant cutting rooting and seed germination rooting. The main mechanism of cutting rooting is cell totipotency and regeneration ability: the cells of cutting branches have been differentiated, but retain totipotency. After the cells at the cutting site are damaged and stimulated, callus is formed by dedifferentiation, and then redifferentiation is carried out to form adventitious root primordia. Seed germination rooting is the pre-formed organ of radicle in seed, which gradually roots. Based on this, it cannot be predicted whether the direct addition of stenotrophomonas maltophilia can effectively promote the cutting rooting of Spatholobus suberectus. SUMMARY

[0006] In view of the deficiencies of the prior art, the present application provides a cutting planting method of Spatholobus suberectus, which can effectively promote the early rooting of Spatholobus suberectus cutting seedlings, improve the overall survival rate, and reduce the cost of Spatholobus suberectus cutting seedling.

[0007] To achieve the above objectives, the present invention provides the following technical solution: A method for propagating *Spatholobus suberectus* by cuttings, the method comprising the following steps: S1. Preparation of cuttings: Cut 15-18cm branches of chicken blood vine, retain 1-2 axillary buds, remove leaves, cut the upper end horizontally and the lower end obliquely, and then soak in disinfectant for disinfection treatment to obtain cuttings for later use. S2. Prepare rooting solution: Prepare rooting solution according to the following formula: 40-60 mL / L of post-fermentation treatment solution of Stenotrophomonas maltophilia + 10-20 mg / L of indoleacetic acid + 120-160 mg / L of salicylic acid + rice slurry water to make up to 1L; The maltophilic oligotrophoblast plant fermentation post-treatment broth is the filtrate obtained by inoculating maltophilic oligotrophoblast into LB liquid medium containing kale powder and bitter chrysanthemum powder, culturing for 15-20 days, and then sterilizing and filtering. S3. Rooting solution soaking treatment: Soak the lower end of the cutting in rooting solution at room temperature for 2-3 hours, and then take it out to obtain the pre-treated cutting. S4. Cutting treatment: Insert the pre-treated cuttings into the sterilized cutting substrate, water thoroughly, disinfect regularly, and wrap the top of the pre-treated cuttings with plastic wrap.

[0008] Preferably, the disinfection treatment in step S1 involves immersing the lower 1 / 5 to 1 / 3 of the sample in a 1-3 mL / L solution of methyl methacrylate and hymexazol for 1-1.5 hours.

[0009] Preferably, the preparation method of the post-fermentation treatment broth of *Stenotrophomonas maltophilia* includes the following steps: S2-1. After drying the Chinese kale and bitter chrysanthemum, pulverize them and pass them through a 20-mesh sieve to obtain Chinese kale powder and bitter chrysanthemum powder; S2-2. Add 10-20 g / L of Chinese kale powder and bitter chrysanthemum powder to LB liquid medium and mix evenly to prepare the culture medium for inoculum. S2-3. Inoculate the above-mentioned culture medium with Stenotrophomonas maltophilia and culture it on a shaker at 120-150 r / min at 28-30℃ for 15-20 days to obtain the fermentation agent for later use. S2-4. After pressing and filtering the above fermentation agent, sterilize the filtrate at 121℃ for 20 minutes to obtain the post-fermentation liquid of Stenotrophomonas maltophilia.

[0010] Preferably, the rice slurry is the filtrate obtained by mixing rice and water at a mass ratio of 1:50-80, grinding them into a slurry, and then filtering it.

[0011] Preferably, the way of soaking in step S3 is to immerse the lower end 1 / 5-1 / 3 of the cutting branch into the rooting liquid.

[0012] Preferably, the formula of the cutting substrate in step S4 is as follows: garden soil 50-60 parts, humic acid 20-30 parts, and perlite 6-10 parts.

[0013] Preferably, the depth of the cutting of the pre-treated cutting branch in S4 is 3-5 cm.

[0014] Preferably, the air humidity after cutting in step S4 is kept at 80%-90%, and the humidity of the cutting substrate is kept such that it can be held in a ball shape and scattered on the ground.

[0015] Preferably, the way of regular disinfection in step S4 is to spray the cutting branch with 800 times liquid of 50% carbendazim wettable powder every 10-15 days.

[0016] The present application provides a cutting planting method of morinda officinalis, which has the following advantages compared with the prior art: The present application adopts the mixture of indole acetic acid, salicylic acid, and rice slurry water as the rooting liquid for the soaking treatment of the lower end of the morinda officinalis cutting branch, wherein the plant fermentation post-treatment liquid of stenotrophomonas maltophilia is the filtrate after sterilization of stenotrophomonas maltophilia cultured in a culture medium added with brassica juncea and bitter aster, which can further promote the early rooting of the morinda officinalis cutting seedling compared with directly using the fermentation liquid of stenotrophomonas maltophilia, and ensure the growth of the cutting seedling, and the addition of brassica juncea and bitter aster in the culture medium further improves the rooting effect of the plant fermentation post-treatment liquid of stenotrophomonas maltophilia on the morinda officinalis cutting, improves the survival rate of the whole cutting, and comprehensively reduces the cost of artificial cutting seedling. BRIEF DESCRIPTION OF DRAWINGS

[0017] Figure 1 It is a schematic diagram of the cutting branch and the rooting liquid in the embodiment of the present application; Figure 2 It is a schematic diagram of the morinda officinalis cutting in the embodiment of the present application; Figure 3 It is a schematic diagram of the rooting of the morinda officinalis cutting in experimental group 1 and experimental group 7 after 20 days; Figure 4 It is a schematic diagram of the rooting of the morinda officinalis cutting in experimental groups 2-6 after 20 days; Figure 5 It is a schematic diagram of the specific rooting of the cutting seedling in experimental group 1 after 20 days. DETAILED DESCRIPTION

[0018] In order to make the purposes, technical solutions and advantages of the embodiments of the present application clearer, the technical solutions in the embodiments of the present application will be described clearly and completely below in conjunction with the embodiments of the present application. Obviously, the described embodiments are some but not all of the embodiments of the present application. Based on the embodiments in the present application, all other embodiments obtained by those of ordinary skill in the art without creative work fall within the protection scope of the present application.

[0019] The Stenotrophomonas maltophilia used below is purchased from Shanghai Fuxiang Biotechnology Co., Ltd. and is ATCC 51331 Stenotrophomonas maltophilia; The mustard powder is obtained by drying fresh mustard at 40°C to a constant weight and then crushing through a 20-mesh sieve; The bitter tea powder is obtained by drying fresh bitter tea at 40°C to a constant weight and then crushing through a 20-mesh sieve; The rice milk water is obtained by mixing rice and water at a mass ratio of 1:70, grinding into a slurry, and then filtering the filtrate.

[0020] Preparation of LB liquid medium: Tryptone: 10 g / L, yeast extract: 5 g / L, sodium chloride: 10 g / L, add distilled water to 1 L; pH value: adjust to 7.0 (adjust using NaOH and HCl) The prepared LB liquid medium is subjected to high-pressure wet heat sterilization at 121°C for 20 minutes to ensure a sterile state. Example 1:

[0021] Preparation of raw materials: 1. Preparation of post-treatment liquid of Stenotrophomonas maltophilia plant fermentation: 1.1. Preparation of post-treatment liquid 1: (1) Add mustard powder and bitter tea powder to the LB liquid medium at 15 g / L, stir uniformly, and use as a strain culture medium; (2) Inoculate Stenotrophomonas maltophilia into the above strain culture medium (inoculation amount is 0.8% of the total mass of the strain culture medium), and culture at 28-30°C and 120 r / min on a shaking table for 18 d to obtain a fermentation agent for standby; (3) Sterilize the filtrate obtained by squeezing the above fermentation agent at 121°C for 20 min to obtain post-treatment liquid 1.

[0022] 1.2. Preparation of post-treatment liquid 2: (1) Add mustard powder to the LB liquid medium at 15 g / L, stir uniformly, and use as a strain culture medium; (2) Stenotrophomonas maltophilia is inoculated into the above-mentioned strain culture medium (inoculation amount is 0.8% of the total mass of the strain culture medium), and cultured at 28-30°C and 120 r / min for 18 days to obtain a fermentation agent for standby; (3) The filtrate of the above-mentioned fermentation agent is sterilized at 121°C for 20 minutes to obtain post-treatment liquid 2.

[0023] 1.3, Preparation of post-treatment liquid 3: (1) 15 g / L of bitterweed powder is added to the LB liquid culture medium and stirred uniformly as a strain culture medium; (2) Stenotrophomonas maltophilia is inoculated into the above-mentioned strain culture medium (inoculation amount is 0.8% of the total mass of the strain culture medium), and cultured at 28-30°C and 120 r / min for 18 days to obtain a fermentation agent for standby; (3) The filtrate of the above-mentioned fermentation agent is sterilized at 121°C for 20 minutes to obtain post-treatment liquid 3.

[0024] 1.4, Preparation of post-treatment liquid 4: (1) Stenotrophomonas maltophilia is inoculated into the LB liquid culture medium (inoculation amount is 0.8% of the total mass of the LB liquid culture medium), and cultured at 28-30°C and 120 r / min for 18 days to obtain a fermentation agent for standby; (2) The filtrate of the above-mentioned fermentation agent is sterilized at 121°C for 20 minutes to obtain post-treatment liquid 4.

[0025] 2, Preparation of Stenotrophomonas maltophilia fermentation liquid: 2.1, Preparation of fermentation liquid 1: (1) 15 g / L of bitterweed powder is added to the LB liquid culture medium and stirred uniformly as a strain culture medium; (2) Stenotrophomonas maltophilia is inoculated into the above-mentioned strain culture medium (inoculation amount is 0.8% of the total mass of the strain culture medium), and cultured at 28-30°C and 120 r / min for 18 days to obtain a fermentation agent for standby; (3) The filtrate of the above-mentioned fermentation agent is sterilized at 121°C for 20 minutes to obtain post-treatment liquid 3.

[0026] 2.2, Preparation of fermentation liquid 2: (1) Stenotrophomonas maltophilia is inoculated into the LB liquid culture medium (inoculation amount is 0.8% of the total mass of the LB liquid culture medium), and cultured at 28-30°C and 120 r / min for 18 days to obtain a fermentation agent for standby; (2) The filtrate of the above-mentioned fermentation agent is sterilized at 121°C for 20 minutes to obtain post-treatment liquid 4.

[0027] 3, Preparation of rooting liquid: Different rooting solutions were prepared according to the following formulations: Formulation of SG-1: 50 mL of post-treatment solution 1 + 15 mg of indole acetic acid + 140 mg of salicylic acid + rice slurry water to make 1 L, SG-1 was obtained; Formulation of SG-2: 50 mL of post-treatment solution 2 + 15 mg of indole acetic acid + 140 mg of salicylic acid + rice slurry water to make 1 L, SG-2 was obtained; Formulation of SG-3: 50 mL of post-treatment solution 3 + 15 mg of indole acetic acid + 140 mg of salicylic acid + rice slurry water to make 1 L, SG-3 was obtained; Formulation of SG-4: 50 mL of post-treatment solution 4 + 15 mg of indole acetic acid + 140 mg of salicylic acid + rice slurry water to make 1 L, SG-4 was obtained; Formulation of SG-5: 50 mL of post-fermentation solution 1 + 15 mg of indole acetic acid + 140 mg of salicylic acid + rice slurry water to make 1 L, SG-5 was obtained; Formulation of SG-6: 50 mL of post-fermentation solution 2 + 15 mg of indole acetic acid + 140 mg of salicylic acid + rice slurry water to make 1 L, SG-6 was obtained; Formulation of SG-7: 15 mg of indole acetic acid + 140 mg of salicylic acid + rice slurry water to make 1 L, SG-7 was obtained. Example 2:

[0028] Method for planting Spatholobus suberectus by cutting: (1) Selecting 1-year-old Spatholobus suberectus woodified branches that are strong and free of pests and diseases, cutting 15-18 cm Spatholobus suberectus branches before new buds sprout, retaining 1-2 axillary buds, removing leaves, and cutting the upper end flat and the lower end obliquely, then immersing the lower end 1 / 5-1 / 3 into a 2 mL / L precision thiophanate-methyl · hymexazol disinfecting solution for 1.5 h to obtain cutting branches for standby; (2) Continuing to immerse the lower end 1 / 5-1 / 3 of the cutting branches into the rooting solution at room temperature for 2.5 h (the cutting branches and the rooting solution are shown in Figure 1 ), then taking out to obtain pre-treated cutting branches; (3) Immersing the pre-treated cutting branches into the disinfecting cutting substrate within 6 h, controlling the insertion depth to be 3-5 cm, then wrapping the upper end with plastic wrap (as shown in Figure 2 ), ensuring the air humidity to be 80%-90%, watering in time, and controlling the cutting substrate humidity to be held together by hand and scattered on the ground; every 10-15 d, spraying the cutting branches with 50% carbendazim wettable powder 800 times liquid for disinfection and disease control. Detection:

[0029] Referring to the cutting propagation method in Example 2 above, garden soil, humic acid, and perlite were mixed evenly in a mass ratio of 55:25:8, and then sterilized at 121°C and 103 kPa for 30 minutes to prepare as the experimental planting substrate. Seven groups of cutting propagation experiments were set up and conducted in a greenhouse with the temperature controlled at 15-25℃. The cuttings were taken on March 10, 2025. The rooting solution used in each group of cutting propagation experiments is shown in Table 1 below: Table 1

[0030] 1. The survival rate of each group was tested at 15, 30, and 60 days after cutting, and the survival rate of each group was calculated. The specific results are shown in Table 2 below: Table 2

[0031] As shown in the table above, the sterilized fermentation solution of Stenotrophomonas maltophilia used in experimental groups 1-4 resulted in a higher overall survival rate of *Spatholobus suberectus* cuttings, and the survival rate did not differ significantly with the addition of different components to the culture medium. In contrast, the unsterilized fermentation solution of Stenotrophomonas maltophilia used in experimental groups 5-6 could improve the survival rate to some extent (compared to experimental group 7), but the improvement was limited and significantly lower than that in experimental groups 1-4.

[0032] 2. The rooting status of *Spatholobus suberectus* in each group was tested 20 days after cutting. Specifically, 5 surviving *Spatholobus suberectus* cuttings were randomly selected from each group 20 days after cutting. The roots were washed with clean water, and the rooting status was observed. The average number of roots (only the main root and effective roots longer than 1 mm were counted) and root length were calculated. The specific results are shown in Table 3 below. Table 3

[0033] As shown in the table above, the post-treatment solution 1 in experimental group 1, which used a culture medium supplemented with both kale powder and chrysanthemum powder, effectively promoted rooting of *Spatholobus suberectus* cuttings and also promoted root growth. This effect was significantly improved compared to adding kale powder or chrysanthemum powder alone to the culture medium. While the inactivated fermentation solution (experimental groups 5-6) showed some improvement in rooting (compared to experimental group 7), the number of roots was lower, resulting in a poorer rooting effect. Specific rooting effects for each group are as follows: Figure 3 and Figure 4 As shown, the specific rooting status of the cuttings in experimental group 1 is as follows: Figure 5 As shown, by Figure 5 It can be seen that in experimental group 1, some lateral roots grew on the main root of the cuttings, and the overall rooting condition was better, which can provide a better nutrient transport channel for the cutting propagation of chicken blood vine.

[0034] The above examples are only used to illustrate the technical solutions of the present application, but not to limit the present application; although the present application has been described in detail with reference to the foregoing examples, those ordinarily skilled in the art should understand: the technical solutions recorded in the foregoing examples can be modified, or some technical features can be replaced equivalently; and these modifications or replacements do not make the essence of the corresponding technical solutions deviate from the spirit and scope of the technical solutions of the embodiments of the present application.

Claims

1. A method for planting a cutting of a Morinda officinalis How, characterized by, The cutting planting method comprises the following steps: S1, preparation of cutting branches: cutting 15-18 cm long branches of Spatholobus suberectus, retaining 1-2 axillary buds, removing leaves, cutting the upper end flat and the lower end obliquely, and then soaking and disinfecting with a disinfectant to obtain cutting branches for standby; S2, preparation of rooting liquid: the rooting liquid is prepared according to the following formula: 40-60 mL / L of post-plant fermentation liquid of stenotrophomonas maltophilia, 10-20 mg / L of indole-3-acetic acid, 120-160 mg / L of salicylic acid, and 1 L of rice slurry water; The post-plant fermentation liquid of stenotrophomonas maltophilia is obtained by inoculating stenotrophomonas maltophilia into LB liquid medium containing mustard powder and bitter tea powder, culturing for 15-20 days, and then sterilizing and filtering to obtain the filtrate; S3, soaking treatment with rooting liquid: soaking the lower end of the cutting branches in the rooting liquid at room temperature for 2-3 hours, and then taking out the pretreated cutting branches; S4, cutting treatment: cutting the pretreated cutting branches into the disinfected cutting substrate, pouring clean water, regularly disinfecting, and wrapping the upper end of the pretreated cutting branches with plastic wrap.

2. The method according to claim 1, wherein the method is characterized by: The disinfection treatment method of the soaking disinfectant in step S1 is to immerse the lower end 1 / 5-1 / 3 into a 1-3 mL / L precision methylthionyl and dodicin disinfectant solution for 1-1.5 hours.

3. The method for propagating *Spatholobus suberectus* by cuttings according to claim 1, characterized in that, The preparation method of the post-plant fermentation liquid of stenotrophomonas maltophilia comprises the following steps: S2-1, drying and crushing mustard and bitter tea through a 20-mesh sieve to obtain mustard powder and bitter tea powder; S2-2, adding 10-20 g / L of the mixed mustard powder and bitter tea powder into the LB liquid medium to uniformly mix as a strain culture medium; S2-3, inoculating stenotrophomonas maltophilia into the strain culture medium, and culturing at 28-30°C and 120-150 r / min on a shaking table for 15-20 days to obtain a fermentation agent for standby; S2-4, sterilizing the filtrate of the fermentation agent at 121°C for 20 minutes to obtain the post-plant fermentation liquid of stenotrophomonas maltophilia.

4. The method according to claim 1, wherein the method is characterized by, The rice slurry water is obtained by mixing rice and water at a mass ratio of 1:50-80, grinding into a slurry, and then filtering the filtrate.

5. The method according to claim 1, wherein the method is characterized by: The soaking method in step S3 is to immerse the lower end 1 / 5-1 / 3 of the cutting branches into the rooting liquid.

6. The method according to claim 1, wherein the method is characterized by: The formula of the cutting substrate in step S4 comprises the following components by weight: 50-60 parts of garden soil, 20-30 parts of humic acid, and 6-10 parts of perlite.

7. The method according to claim 1, wherein the method is characterized by: The depth of the cutting of the pretreated cutting branches in S4 is 3-5 cm.

8. The method according to claim 1, wherein the method is characterized by: In step S4, the air humidity is maintained at 80%-90% after cutting, and the humidity of the cutting substrate is maintained by holding the cutting substrate in a lump and scattering it on the ground.

9. The method according to claim 1, wherein the method is characterized by: The regular disinfection method in step S4 is to spray and disinfect the cutting branches with 800 times liquid of 50% carbendazim wettable powder every 10-15 days.

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