Cultivation method for reducing content of mimosine in mimosa pudica

Abstract

The invention belongs to the technical field of cultivation, and provides a cultivation method for reducing the content of mimosine in mimosa pudica, which comprises the steps of mimosa pudica seed pretreatment, culture medium preparation, growth environment regulation and control, fertilizer management and the like. According to the mimosa pudica obtained through the cultivation method, the content of mimosine is obviously reduced compared with that of mimosa pudica obtained through a traditional cultivation method, and the method has important significance for widening the application scenes of the mimosa pudica in the fields of ornamental plants, forage grass and the like; the cultivation method is low in cost and suitable for large-scale popularization.

Description

Technical Field

[0001] This invention belongs to the field of cultivation technology, specifically relating to a cultivation method for reducing the content of mimosine in Mimosa pudica. Background Technology

[0002] Plants in the genus *Mimosa* are shrubs or herbaceous plants, occasionally trees or climbers, and are usually thorny. There are approximately 500 species, mainly distributed in tropical America. According to *Flora of China* (2010), there are currently 3 species and 2 varieties in China, namely *Mimosa pudica* (…). Mimosa pudica ), Mimosa pudica ( Mimosa bimucronata ), Brazilian mimosa ( Mimosa diplotricha ), Brazilian Mimosa (original variety) Mimosa diplotricha var. diplotricha ) and thornless Brazilian mimosa ( Mimosa diplotricha var. inermis ).

[0003] Mimosa pudica is a perennial leguminous plant that typically lives in symbiosis with rhizobia, fixing atmospheric nitrogen to improve soil fertility. With its well-developed root system and strong drought resistance, it is an excellent plant for sand control, soil stabilization, and water conservation. It is also rich in plant protein, making it a potential low-input leguminous forage crop. Furthermore, due to the rapid closure of its leaves in response to mechanical stimulation, Mimosa pudica is often used as an ornamental plant or for science education. However, the core toxic component of Mimosa pudica—mimosine—is a secondary metabolite. This toxicity typically increases in the plant's defensive response to environmental stresses (such as light imbalance, nutrient deficiency, mechanical damage, and pests and diseases). It is clearly toxic to humans and animals, causing hair loss and skin irritation with prolonged contact, and nausea, vomiting, and liver and kidney damage after ingestion. This limits its application as an ornamental plant and forage crop.

[0004] Existing technologies only document the regulation of mimosine in other plants: Leucaena leucocephala ( Leucaena leucocephala*Leucaena leucaena* is a small tree belonging to the genus *Leucaena* in the legume family. Its plant contains the secondary metabolite mimosine. The regulation of mimosine content in *Leucaena leucaena* mainly involves the following aspects: Rodrigues-Corrêa et al. (2018) disclosed the effects of various stress-related secondary metabolite inducers and acute ultraviolet exposure on mimosine accumulation in *Leucaena leucaena* seedlings. The results showed that the concentration of mimosine increased in the roots and aboveground parts of seedlings treated with 84 ppm jasmonic acid (JA) and 10 ppm ketene, as well as in the aboveground parts treated with UV-C radiation. Hond et al. (2019) disclosed the mimosine content in various *Leucaena leucaena* tissues exposed to different environmental conditions. Their research showed that the mimosine content in leaves of *Leucaena leucaena* significantly increased under strong light, high temperature, low temperature, drought, salt stress, and high nitrogen supply; while under suitable temperature, sufficient water supply, weak light or darkness treatment, and moderately low nitrogen conditions, the mimosine content significantly decreased.

[0005] Although Pillathil et al. (2024) disclosed that the concentration of active compounds, including phenols, flavonoids, alkaloids and saponins, in Mimosa pudica increased when exposed to UV-B radiation, they did not address the effect of UV-B radiation on the content of mimosine in Mimosa pudica.

[0006] In summary, there is no existing technical solution for reducing the mimosine content in Mimosa pudica plants. Therefore, developing a simple, scalable cultivation method to reduce the mimosine content in Mimosa pudica is essential for addressing its application prospects in garden plants and agriculture. Summary of the Invention

[0007] The purpose of this invention is to provide a cultivation method for reducing the mimosine content in Mimosa pudica, which can significantly reduce the mimosine content in Mimosa pudica.

[0008] The present invention achieves the above objectives using the following methods:

[0009] A cultivation method for reducing the mimosine content in Mimosa pudica includes the following steps: soaking Mimosa pudica seeds in an effective amount of Mimosa pudica extract, and then planting the treated seeds into plants.

[0010] The above-mentioned mimosa seeds should be selected from healthy and plump ones. The methods include, but are not limited to, visual screening, winnowing, water selection, and gravity cleaning. As long as impurities and shriveled seeds can be removed from the mimosa seeds to obtain healthy and plump seeds, it is acceptable.

[0011] The mimosa extract is prepared by soaking mimosa seeds in an aqueous solution, with an effective mass concentration of 0.01% to 30%.

[0012] Furthermore, the effective concentration of the aqueous solution of the mimosa extract is 0.05%~30%; the effective concentration of the aqueous solution of the mimosa extract is 0.1%~30%; the effective concentration of the aqueous solution of the mimosa extract is 0.5%~25%; the effective concentration of the aqueous solution of the mimosa extract is 1%~20%; the effective concentration of the aqueous solution of the mimosa extract is 1%~15%; the effective concentration of the aqueous solution of the mimosa extract is 1%~10%; and the effective concentration of the aqueous solution of the mimosa extract is 1%~5%.

[0013] Furthermore, the effective concentration of the aqueous solution of the mimosa extract includes, but is not limited to, 0.2%, 0.7%, 1%, 1.5%, 2%, 2.1%, 2.2%, 2.3%, 2.4%, 2.5%, 2.6%, 2.7%, 2.8%, 2.9%, 3%, 3.1%, 3.2%, 3.3%, 3.4%, 3.5%, 3.6%, 3.7%, 3.8%, 3.9%, 4%, 4.1%, 4.2%, 4.3%, 4.4%, 4.5%, 4.6%, 4.7%, 4.8%, 4.9%, 5%, 5.1%, 5.2%, 5.3%, 5.4%, 5.5%, 5.6%, 5.7%, 5.8%, 5.9%, 6%, 6.5%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, and 30%.

[0014] During the process of soaking the mimosa seeds with the mimosa extract, the soaking temperature is 20~30℃ and the soaking time is 6~8h.

[0015] The "extract" described in this invention refers to a substance formed by using *Mimosa pudica* as raw material, and through a physicochemical extraction and separation process to selectively obtain and / or concentrate one or more target components (i.e., components that reduce the mimosine content in *Mimosa pudica*) from the raw material, based on the need to reduce the mimosine content in the final extracted substance. This includes extraction methods known in the art and combinations thereof; extraction methods include, but are not limited to, solvent (water, organic solvent) treatment, solid-phase extraction, supercritical CO2, thermal, ultrasonic extraction, exposure to pressure or centrifugal force, or other chemical and / or mechanical processes to obtain the target component from the raw material.

[0016] The organic solvents include, but are not limited to, alcohols, ethyl acetate, acetone, and oils.

[0017] The alcohols include, but are not limited to, methanol, ethanol, and isopropanol.

[0018] The extracted mimosa plants include, but are not limited to, the whole plant, above-ground parts, underground parts, leaves, branches, flowers, and seeds.

[0019] The preparation method of the mimosa extract includes the following steps: pulverizing mimosa seeds, soaking them in ethanol for 24-48 hours, filtering, and then evaporating the filtrate under negative pressure until it becomes viscous; wherein the ratio of mimosa seeds to ethanol (volume ratio) is 1: (1-5), and the evaporation temperature is 30-65℃.

[0020] The above cultivation method also includes planting the soaked Mimosa seeds in a cultivation substrate containing molybdate.

[0021] The cultivation substrate can be a mixture of organic substrate, inorganic substrate and fertilizer in a volume ratio of 5:(1~2):(1~2):1, and then mixed with molybdate.

[0022] The molybdate content is 0.01% to 0.1% by mass; specifically, it can be 0.01%, 0.02%, 0.03%, 0.04%, 0.05%, 0.06%, 0.07%, 0.08%, 0.09% and / or 0.1%.

[0023] The molybdates include, but are not limited to, calcium molybdate, zinc molybdate, sodium molybdate, and ammonium molybdate.

[0024] The organic matrix is ​​a humic matrix, including but not limited to humus soil, peat, humic acid matrix, etc.

[0025] The inorganic matrix is ​​a porous inorganic matrix, including but not limited to perlite, vermiculite, pumice, volcanic rock, zeolite, etc.

[0026] The fertilizer may be organic or inorganic; the organic fertilizer is well-rotted organic fertilizer, including but not limited to well-rotted oilseed cakes, livestock and poultry manure, plant residues / compost, and agricultural by-products; further, the well-rotted oilseed cakes include but are not limited to well-rotted soybean meal, rapeseed meal, cottonseed meal, peanut meal, sesame meal, and castor meal.

[0027] The pH value of the cultivation substrate can be adjusted to 6.5~7.0.

[0028] During the planting process, the soaked mimosa seeds are scattered on the surface of the cultivation substrate, and then the surface is covered with soil.

[0029] The cultivation substrate can be watered first, and then the soaked mimosa seeds can be sown.

[0030] The thickness of the surface soil cover is 0.5~1cm.

[0031] The above cultivation methods may also include applying microbial fertilizer to the roots of the mimosa or to the cultivation substrate.

[0032] The microbial fertilizer is composed of the following raw materials in parts by weight: fiber microorganisms ( Cellulosimicrobium cellulans 18-22 parts of fermentation broth, Bacillus subtilis ( Bacillus subtilis 18-22 parts of fermentation broth, Bacillus pumilus ( Bacillus pumilus 16-18 parts fermentation broth, 9-13 parts urea, 9-13 parts dipotassium hydrogen phosphate, and 18-22 parts water.

[0033] The method for preparing the microbial fertilizer includes the following steps:

[0034] (1) Fiber microbes, Bacillus subtilis and Bacillus pumilus were activated and cultured separately;

[0035] (2) The activated Bacillus subtilis, Bacillus pumilus and cellulose microbes were inoculated into LB medium at an inoculation amount of 4-6% v / v and fermented separately at a temperature of 35-37℃ for 36-60h to obtain fermentation broth of Bacillus subtilis, Bacillus pumilus and cellulose microbes.

[0036] (3) Mix Bacillus subtilis, Bacillus pumilus and cellulose microorganism fermentation broth with urea, dipotassium hydrogen phosphate and water evenly to obtain microbial fertilizer.

[0037] The effective live bacteria count in the microbial fertilizer is ≥0.5 billion / ml.

[0038] The microbial fertilizer needs to be diluted with 50 to 100 times the amount of water before application.

[0039] The diluted microbial fertilizer solution should be sprayed when the mimosa has grown to 4 leaves, at a rate of 4.5-10 L / mu, and every 20-30 days.

[0040] In addition to microbial fertilizer, foliar fertilizer can also be applied. The foliar fertilizer should be sprayed once each during the seedling stage, growth stage, and budding stage, after 16:00, and the leaves should be moist but not dripping.

[0041] The foliar fertilizer may be potassium dihydrogen phosphate, with a spraying amount of 0.03 wt%.

[0042] The above greenhouse management is routine management, with the temperature controlled between 10 and 35°C, the air humidity controlled between 50% and 70%, and natural light and ventilation maintained for more than 6 hours a day.

[0043] The above methods can be used to reduce the content of mimosine in Mimosa pudica.

[0044] The beneficial effects of this invention are as follows:

[0045] (1) This invention provides a cultivation method to reduce the content of mimosine in Mimosa pudica, which can significantly reduce the content of mimosine in Mimosa pudica and broaden the application scenarios of Mimosa pudica in ornamental plants, forage grass and other fields; and the cultivation method is low in cost and suitable for large-scale promotion.

[0046] (2) Treatment of mimosa seeds with mimosa extract inhibited the synthesis of mimosine from the start.

[0047] (3) In this invention, sodium molybdate was added to the cultivation substrate and the content of mimosine in Mimosa pudica decreased. It is speculated that molybdenum may have reduced the allocation of nitrogen to secondary metabolites such as mimosine.

[0048] (4) By applying the microbial fertilizer prepared in this invention, the content of mimosine in Mimosa pudica can be reduced. Attached Figure Description

[0049] Figure 1 The graph shows a comparison of the mimosine content in *Mimosa pudica* cultivated using traditional methods and the method of this invention; where: a is a chromatogram of a mimosine standard sample, b is a chromatogram of *Mimosa pudica* extract obtained using the cultivation method in Example 2, and c is a chromatogram of *Mimosa pudica* extract obtained using the traditional cultivation method in Comparative Example 5. Detailed Implementation

[0050] The technical solutions in the embodiments of the present invention will be clearly and completely described below. The described embodiments are only a part of the present invention, and not the entirety of the invention. 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.

[0051] A cultivation method for reducing mimosine in Mimosa pudica includes the following steps:

[0052] (1) Seed pretreatment

[0053] ① Select healthy and plump mimosa seeds for later use;

[0054] ② Prepare a 0.01%~30% (w / w) solution of mimosa extract, soak the selected seeds in the mimosa extract solution at a temperature of 20~30℃ for 6~8 hours, and then filter.

[0055] (2) Preparation of cultivation substrate

[0056] Mix humus, perlite, vermiculite and well-rotted soybean meal in a volume ratio of 5:(1~2):(1~2):1 evenly, then add sodium molybdate with a mass percentage of 0.02%, mix evenly, and adjust the pH to 6.5~7.0 to obtain the cultivation substrate.

[0057] (3) Planting

[0058] Soak the mimosa seeds and scatter them on the surface of the cultivation substrate, then cover the surface with 0.5-1cm of soil.

[0059] (4) Management of growth environment

[0060] ① Temperature Management

[0061] The temperature should be controlled between 10 and 35℃;

[0062] ② Lighting Management

[0063] More than 6 hours of natural sunlight per day;

[0064] ③Gas regulation

[0065] Natural ventilation;

[0066] (5) Fertilizer Management

[0067] ① Foliar fertilizer

[0068] Foliar fertilizer was sprayed once each during the seedling stage, growth stage, and budding stage; the foliar fertilizer consisted of the following components: 0.03wt% potassium dihydrogen phosphate;

[0069] ② Microbial fertilizer

[0070] When the mimosa plant has grown to 4 leaves, start spraying with diluted microbial fertilizer solution, once every 20-30 days, at a rate of 4.5-10 L / mu. The microbial fertilizer is composed of the following raw materials in parts by weight: 18-22 parts of fiber microbial fermentation liquid, 18-22 parts of Bacillus subtilis fermentation liquid, 16-18 parts of Bacillus pumilus fermentation liquid, 9-13 parts of urea, 9-13 parts of dipotassium hydrogen phosphate, and 18-22 parts of water.

[0071] The microbial fertilizer needs to be diluted with 50 to 100 times the amount of water before application.

[0072] The method for preparing the microbial fertilizer includes the following steps:

[0073] (1) Fiber microbes, Bacillus subtilis and Bacillus pumilus were activated and cultured separately;

[0074] (2) The activated Bacillus subtilis, Bacillus pumilus and cellulose microbes were inoculated into LB medium at an inoculation amount of 4-6% v / v and fermented separately at a temperature of 35-37℃ for 36-60h to obtain fermentation broth of Bacillus subtilis, Bacillus pumilus and cellulose microbes.

[0075] (3) Mix Bacillus subtilis, Bacillus pumilus and cellulose microorganism fermentation broth with urea, dipotassium hydrogen phosphate and water evenly to obtain microbial fertilizer.

[0076] The preparation method of the mimosa extract includes the following steps: pulverizing mimosa seeds, soaking them in ethanol for 30 hours, filtering, and then evaporating the filtrate under negative pressure until it becomes viscous; wherein the ratio of mimosa seeds to ethanol (volume ratio) is 1:3, and the evaporation temperature is 30~65℃.

[0077] Example 1

[0078] Cultivation location: Shenyang City, Liaoning Province, ordinary greenhouse.

[0079] Cultivation can begin in mid-September.

[0080] Cultivation steps:

[0081] S1, Seed Pretreatment

[0082] S1-1. Soak Mimosa pudica seeds in water, remove floating seeds, and keep the seeds that sink to the bottom for later use;

[0083] S1-2. Prepare a 1% (w / w) solution of mimosa extract, soak the submerged seeds in the mimosa extract solution at a temperature of 20-30°C for 8 hours, and filter to obtain mimosa seeds 1.

[0084] S2, Preparation of cultivation substrate

[0085] Mix humus, perlite, vermiculite, and well-rotted soybean meal in a volume ratio of 5:2:2:1 until homogeneous. Then add sodium molybdate at a mass fraction of 0.02%, mix well, and adjust the pH to 6.5-7.0 to obtain the cultivation substrate.

[0086] S3, Planting

[0087] Spread the cultivation substrate into the cultivation bed and water it; scatter the mimosa seeds on the surface of the cultivation substrate and cover the surface with 0.5-1cm of soil;

[0088] S4. Growth Environment Management

[0089] S4-1 Temperature and Humidity Management

[0090] Temperature is controlled at 10~35℃, air humidity is controlled at 50%~70%, and drip irrigation is used;

[0091] S4-2, Light Management

[0092] More than 6 hours of natural sunlight per day;

[0093] S4-3, Ventilation Management

[0094] Ventilation will be turned on from 10:30 to 14:00;

[0095] S5, Fertilizer Management

[0096] Foliar fertilizer: Spray foliar fertilizer once each during the seedling stage, growth stage, and before bud formation; the foliar fertilizer is 0.03wt% potassium dihydrogen phosphate; prune diseased and weak branches.

[0097] Microbial fertilizer: When the mimosa has grown to 4 leaves, start spraying the diluted microbial fertilizer solution once every 30 days, with an application rate of 4.5L / acre.

[0098] Microbial fertilizer is composed of the following raw materials in parts by weight: 20 parts of fiber microbial fermentation liquid, 20 parts of Bacillus subtilis fermentation liquid, 18 parts of Bacillus pumilus fermentation liquid, 11 parts of urea, 11 parts of dipotassium hydrogen phosphate, and 20 parts of water.

[0099] The method for preparing the microbial fertilizer includes the following steps:

[0100] (1) Fiber microbes, Bacillus subtilis and Bacillus pumilus were activated and cultured separately;

[0101] (2) The activated Bacillus subtilis, Bacillus pumilus and cellulose microbes were inoculated into LB medium at an inoculation amount of 4-6% v / v and fermented at 37℃ for 48h to obtain fermentation broths of Bacillus subtilis, Bacillus pumilus and cellulose microbes respectively.

[0102] (3) Mix Bacillus subtilis, Bacillus pumilus and cellulose microorganism fermentation broth with urea, dipotassium hydrogen phosphate and water evenly to obtain microbial fertilizer.

[0103] The above-mentioned microbial fertilizer needs to be diluted with 50 times the amount of water.

[0104] The fibroblasts, Bacillus subtilis, and Bacillus pumilus mentioned were all purchased commercially.

[0105] The preparation method of the mimosa extract includes the following steps: pulverizing mimosa seeds, soaking them in ethanol for 30 hours, filtering, and then evaporating the filtrate under negative pressure until it becomes viscous; wherein the ratio of mimosa seeds to ethanol (volume ratio) is 1:3, and the evaporation temperature is 30~65℃.

[0106] Example 2

[0107] Cultivation location: Shenyang City, Liaoning Province, ordinary greenhouse.

[0108] Cultivation can begin in mid-September.

[0109] Cultivation steps:

[0110] S1, Seed Pretreatment

[0111] S1-1. Soak the seeds of Mimosa pudica in water, remove the floating seeds, and keep the seeds that sink to the bottom for later use;

[0112] S1-2. Prepare a 3% (w / w) solution of mimosa extract, soak the submerged seeds in the mimosa extract solution at a temperature of 20-30°C for 6 hours, and filter to obtain mimosa seeds 1.

[0113] S2, Preparation of cultivation substrate

[0114] Mix humus, perlite, vermiculite, and well-rotted soybean meal in a volume ratio of 5:2:2:1 until homogeneous. Then add sodium molybdate at a mass fraction of 0.02%, mix well, and adjust the pH to 6.5-7.0 to obtain the cultivation substrate.

[0115] S3, Planting

[0116] Spread the cultivation substrate into the cultivation bed and water it; scatter the mimosa seeds on the surface of the cultivation substrate and cover the surface with 0.5-1cm of soil;

[0117] S4. Growth Environment Management

[0118] S4-1 Temperature and Humidity Management

[0119] Temperature is controlled at 10~35℃, air humidity is controlled at 50%~70%, and drip irrigation is used;

[0120] S4-2, Light Management

[0121] More than 6 hours of natural sunlight per day;

[0122] S4-3, Ventilation Management

[0123] Ventilation will be turned on from 10:30 to 14:00;

[0124] S5, Fertilizer Management

[0125] Foliar fertilizer: Spray foliar fertilizer once each during the seedling stage, growth stage, and before bud formation; the foliar fertilizer is 0.03wt% potassium dihydrogen phosphate; prune diseased and weak branches.

[0126] Microbial fertilizer: Start spraying microbial fertilizer when the mimosa plant has grown to 4 leaves, spraying once every 25 days, at a rate of 10L / acre. The preparation methods for the microbial fertilizer and the mimosa extract are described in Example 1.

[0127] Example 3

[0128] Cultivation location: Shenyang City, Liaoning Province, ordinary greenhouse.

[0129] Cultivation time: Mid-September.

[0130] S1, Seed Pretreatment

[0131] S1-1. Soak the seeds of Mimosa pudica in water, remove the floating seeds, and keep the seeds that sink to the bottom for later use;

[0132] S1-2. Prepare a 5% (w / w) solution of mimosa extract, soak the submerged seeds in the mimosa extract solution at a temperature of 20-30°C for 6 hours, and filter to obtain mimosa seeds 1.

[0133] S2, Preparation of cultivation substrate

[0134] Mix humus, perlite, vermiculite, and well-rotted soybean meal in a volume ratio of 5:2:2:1 until homogeneous. Then add sodium molybdate at a mass fraction of 0.02%, mix well, and adjust the pH to 6.5-7.0 to obtain the cultivation substrate.

[0135] S3, Planting

[0136] Spread the cultivation substrate into the cultivation bed and water it; scatter 1 seed of Mimosa pudica on the surface of the cultivation substrate, and then cover the surface with 0.5-1cm of soil;

[0137] S4. Growth Environment Management

[0138] S4-1 Temperature and Humidity Management

[0139] Temperature is controlled at 10~35℃, air humidity is controlled at 50%~70%, and drip irrigation is used;

[0140] S4-2, Light Management

[0141] More than 6 hours of natural sunlight per day;

[0142] S4-3, Ventilation Management

[0143] Ventilation will be turned on from 10:30 to 14:00;

[0144] S5, Fertilizer Management

[0145] Foliar fertilizer: Spray foliar fertilizer once each during the seedling stage, growth period, and before bud formation; the foliar fertilizer is 0.03wt% potassium dihydrogen phosphate, with the remainder being water; prune diseased and weak branches.

[0146] Microbial fertilizer: Start spraying microbial fertilizer when the mimosa has grown to 4 leaves, spray once every 20 days, at a rate of 4.5L / acre.

[0147] The preparation methods of the microbial fertilizer and mimosa extract are described in Example 1.

[0148] Comparative Example 1

[0149] Except for step S1, in which the mimosa seeds are soaked in mimosa extract, all other steps are the same as in Example 2.

[0150] Comparative Example 2

[0151] Except for the absence of sodium molybdate in step S2, all other steps are the same as in Example 2.

[0152] Comparative Example 3

[0153] Except for the absence of microbial fertilizer in step S5, all other steps are the same as in Example 2.

[0154] Comparative Example 4

[0155] Except for the absence of sodium molybdate in step S2 and the application of microbial fertilizer in step S5, all other steps are the same as in Example 2.

[0156] Comparative Example 5: Cultivation of Mimosa pudica using traditional greenhouse methods

[0157] Cultivation location: Shenyang City, Liaoning Province, ordinary greenhouse.

[0158] Cultivation can begin in mid-September.

[0159] Cultivation steps:

[0160] S1, Seed Pretreatment

[0161] Soak the seeds of the mimosa plant in water, remove the floating seeds, and keep the seeds that sink to the bottom for later use;

[0162] S2, Preparation of cultivation substrate

[0163] Mix humus, perlite, vermiculite, and well-rotted soybean meal in a volume ratio of 5:2:2:1 evenly, and adjust the pH to 6.5-7.0 to obtain the cultivation substrate.

[0164] S3, Planting

[0165] Spread the cultivation substrate into the cultivation bed and water it; scatter the sunken seeds on the surface of the cultivation substrate and cover the surface with 0.5-1cm of soil;

[0166] S4. Growth Environment Management

[0167] S4-1 Temperature and Humidity Management

[0168] Temperature is controlled at 10~35℃, air humidity is controlled at 50%~70%, and drip irrigation is used;

[0169] S4-2, Light Management

[0170] More than 6 hours of natural sunlight per day;

[0171] S4-3, Ventilation Management

[0172] Ventilation will be turned on from 10:30 to 14:00.

[0173] S5, Fertilizer Management

[0174] Foliar fertilizer: Spray foliar fertilizer once each during the seedling stage, growth stage, and before bud formation; the foliar fertilizer is 0.03wt% potassium dihydrogen phosphate; prune diseased and weak branches.

[0175] In each of the above cases, three duplicate plots were set up for planting; after the offspring were grown, the above-ground parts of the mimosa were sampled from each plot using the five-point sampling method.

[0176] The source of the mimosine standard sample was: commercially available.

[0177] Mimosine detection method: The mimosine standard solution is dissolved in water to prepare a 1 mg / mL aqueous solution, and then diluted stepwise as needed.

[0178] Extraction method of Mimosa pudica: Take the aerial parts separately, dry them in the shade and then crush them. Accurately weigh 5.0 g (accurate to 0.01 g) into a 250 mL stoppered conical flask, add 50 mL of 50% methanol-water mixture, extract by sonication for 30 min, cool and centrifuge at 4200 r / min for 5 min, take the supernatant and filter it through a 0.22 μm organic filter membrane for liquid chromatography analysis.

[0179] Content determination method: An Agilent 1260 high performance liquid chromatograph was used with a C18 column as the stationary phase. The mobile phase consisted of methanol and 10 mM sodium octanesulfonate solution (adjusted to pH 4.0 with phosphoric acid) (10 / 90, v / v). The flow rate was 0.8 mL / min, the detection wavelength was 280 nm, the column temperature was 30 ℃, and the external standard method was used for quantitative analysis.

[0180] Table 1 Comparison of mimosine Case Example 1 Example 2 Example 3 Comparative Example 1 Comparative Example 2 Comparative Example 3 Comparative Example 4 Comparative Example 5 Mimosine (mg / kg) 111.9±8.2 15.0±1.8 23.8±2.6 237.8±23.3 59.2±4.8 36.2±1.9 74.4±6.4 318.2±33.1

[0181] Figure 1 This is a comparison chart of the mimosine content in *Mimosa pudica* cultivated using the conventional method (Comparative Example 5) and the method of the present invention (Example 2). From... Figure 1 It can be seen that the mimosine content of the Mimosa obtained by the cultivation method of this invention is significantly lower than that obtained by the traditional method.

Claims

1. A cultivation method for reducing the mimosine content in Mimosa pudica, characterized in that, Includes the following steps: Mimosa seeds were soaked in mimosa extract, and the treated seeds were then planted.

2. The method according to claim 1, characterized in that, The mimosa extract is obtained by soaking mimosa seeds in an aqueous solution with a mass concentration of 0.01% to 30%.

3. The method according to claim 1 or 2, characterized in that, The preparation method of the mimosa extract includes the following steps: pulverizing mimosa seeds, soaking them in ethanol for 24-48 hours, filtering, and then evaporating the filtrate under negative pressure until it becomes viscous; wherein the volume ratio of mimosa seeds to ethanol is 1: (1-5), and the evaporation temperature is 30-65℃.

4. The method according to claim 1 or 2, characterized in that, The soaking temperature is 20~30℃, and the soaking time is 6~8h.

5. The method according to claim 1, characterized in that, The cultivation method also includes planting the soaked mimosa seeds in a cultivation substrate containing molybdate.

6. The method according to claim 5, characterized in that, The molybdate content is 0.01% to 0.1% by mass.

7. The method according to claim 1 or 5, characterized in that, The cultivation method also includes applying microbial fertilizer to the roots of the mimosa or to the cultivation substrate.

8. The method according to claim 7, characterized in that, The microbial fertilizer is composed of the following raw materials in parts by weight: 18-22 parts of fiber microbial fermentation liquid, 18-22 parts of Bacillus subtilis fermentation liquid, 16-18 parts of Bacillus pumilus fermentation liquid, 9-13 parts of urea, 9-13 parts of dipotassium hydrogen phosphate, and 18-22 parts of water.

9. The application of the method according to any one of claims 1 to 8 in reducing the mimosine content in Mimosa pudica.

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