Mimosa extract plant growth stimulating hormone, preparation method and application

Abstract

This invention discloses a plant growth stimulant derived from *Mimosa pudica* extract, its preparation method, and its application. The *Mimosa pudica* extract is used to prepare a plant growth stimulant. The plants mentioned include pepper and tomato. The plant growth stimulant is used to promote crop growth, increase yield, improve quality, and / or enhance stress resistance. The *Mimosa pudica* extract contains 2,4-di-tert-butylphenol, 5-hydroxytryptamine, and mimosine. The plant growth stimulant comprises an extract of *Mimosa pudica*, wherein the *Mimosa pudica* extract has an active ingredient content of 15%–20%. The advantages of this invention are that it can improve plant resistance, promote plant growth, and improve fruit quality.

Description

Technical Field

[0001] This invention belongs to the field of plant nutrition technology, specifically relating to a plant growth stimulant extracted from Mimosa pudica, its preparation method, and its application. Background Technology

[0002] With the increasing intensification of global agriculture, the long-term irrational application of chemical fertilizers and pesticides has led to a series of severe challenges, such as declining agricultural product quality, increased pathogen resistance, soil acidification, salinization, and soil health problems including imbalances in microbial community diversity. How to restore degraded soil and reduce environmental burden while ensuring high crop yields has become a core scientific issue restricting sustainable agricultural development. To alleviate dependence on chemical inputs, biostimulants are one of the green solutions that have emerged to address these problems. Unlike traditional fertilizers, biostimulants do not directly provide nutrients but activate the plant's own potential by regulating plant physiological metabolism and rhizosphere microecology. Among them, plant-derived biostimulants have become a current research hotspot due to their natural origin, low toxicity, low likelihood of inducing resistance, and high environmental compatibility. Currently, commercially available biostimulants are mainly derived from seaweed extracts, humic acid, and protein hydrolysates, but their active ingredients are complex and their mechanisms of action are unclear, limiting their precise application.

[0003] Mimosa pudica ( Mimosa pudica Mimosa pudica (L.) is a perennial herbaceous plant belonging to the legume family. As a unique plant resource rich in flavonoids, phenolic acids, and triterpenoids, it possesses extremely strong biological activity. Current research mainly focuses on its medicinal value, while its application potential in the agricultural field has not yet been fully explored. In particular, the biostimulatory effects of Mimosa pudica extract and its physiological regulatory mechanisms on crops remain a blank.

[0004] Therefore, developing a novel plant-derived biostimulant based on Mimosa pudica extract aligns with the national strategy of "double reduction" and green pest control, provides key technical support for achieving environmentally friendly, efficient, and intensive agricultural development, has broad application prospects and market value, and is of great significance for promoting the research and development of novel biostimulants. Summary of the Invention

[0005] The purpose of this invention is to provide a plant growth stimulant extracted from Mimosa pudica, its preparation method, and its application, which has a growth-promoting effect and improves crop quality.

[0006] To achieve the above objectives, the technical solution adopted by the present invention includes:

[0007] Applications of Mimosa pudica extract in the preparation of plant growth stimulants.

[0008] Optionally, the plants include peppers and tomatoes, and the plant growth stimulants are used to promote crop growth, increase yield, improve quality, and / or increase stress resistance.

[0009] Optionally, the mimosa extract contains 2,4-di-tert-butylphenol, 5-hydroxytryptamine, and mimosine.

[0010] Optionally, the method for preparing the mimosa extract includes: After drying and pulverizing the whole plant or aerial parts of Mimosa pudica, extract it using water, ethanol aqueous solution or methanol aqueous solution as extraction solvent at 40-80℃ for 1-4 hours. After filtration, collect the extract and concentrate it under reduced pressure to obtain Mimosa pudica extract.

[0011] Optionally, the extraction solvent is an aqueous ethanol solution with a volume concentration of 60% to 80%, and the material-to-liquid ratio is 1:8 to 1:15 g / mL.

[0012] Optionally, the extraction method is ultrasonic-assisted extraction or heating reflux extraction, with an extraction temperature of 50–70°C and an extraction time of 2–3 hours.

[0013] A plant growth stimulant extracted from Mimosa pudica, wherein the plant growth stimulant is in the form of a soluble liquid. The soluble liquid comprises 15%–20% mimosa extract, 3%–6% dimethyl sulfoxide, 7%–14% ethanol, 10% NP-10, and water to make up to 100%.

[0014] A method for preparing plant growth stimulants from Mimosa pudica extract includes: After drying and pulverizing the whole plant or aerial parts of Mimosa pudica, extracting it with water, ethanol aqueous solution or methanol aqueous solution at 40-80℃ for 1-4 hours, collecting the extract after filtration, and concentrating it under reduced pressure to obtain Mimosa pudica extract. According to the formula, the obtained mimosa extract was prepared into a soluble liquid.

[0015] Optionally, the extraction solvent is an aqueous ethanol solution with a volume concentration of 60% to 80%, and the material-to-liquid ratio is 1:8 to 1:15 g / mL.

[0016] Optionally, the extraction method is ultrasonic-assisted extraction or heating reflux extraction, with an extraction temperature of 50–70°C and an extraction time of 2–3 hours.

[0017] The advantages of this invention are: ① While there are numerous reports on biostimulants, research on plant-derived biostimulants accounts for a relatively small proportion. Therefore, the development of novel, environmentally friendly, and highly effective plant-derived biostimulants is of great significance.

[0018] ②Exogenous application of plant growth stimulants from Mimosa pudica extract showed significant growth-promoting and resistance-inducing activities in crops, which can better serve agricultural production.

[0019] ③ The extract of Mimosa pudica and its preparations can be rapidly degraded by microorganisms in the natural environment, with no risk of residual accumulation. They will not cause soil compaction or groundwater pollution, and are conducive to maintaining the structure of the rhizosphere microbial community and soil health. Attached Figure Description

[0020] The accompanying drawings are provided to further understand the present disclosure and form part of the specification. They are used together with the following detailed description to explain the present disclosure, but do not constitute a limitation thereof.

[0021] Figure 1 The effect of mimosa extract plant growth stimulants on chlorophyll a and chlorophyll b in pepper; Figure 2 The effect of mimosa extract plant growth stimulants on malondialdehyde content in capsicum. Figure 3 The effect of mimosa extract plant growth stimulants on tomato chlorophyll a and chlorophyll b; Figure 4 The effect of mimosa extract plant growth stimulants on malondialdehyde content in tomatoes. Detailed Implementation

[0022] The essential features of the present invention can be seen from the following embodiments, but these embodiments are for illustrative purposes only and are not intended to limit this aspect.

[0023] The application of mimosa extract in the preparation of plant growth stimulants; plants include pepper and tomato, and the plant growth stimulants are used to promote crop growth, increase yield, improve quality and / or increase stress resistance. Mimosa extract contains 2,4-di-tert-butylphenol, 5-hydroxytryptamine and mimosine.

[0024] The preparation method of mimosa extract includes: drying and pulverizing the whole herb or aerial parts of mimosa, extracting with water, ethanol aqueous solution or methanol aqueous solution as extraction solvent, extracting at 40-80℃ for 1-4 hours, collecting the extract after filtration, and concentrating under reduced pressure to obtain mimosa extract paste.

[0025] Yes, the extraction solvent is an ethanol-water solution with a volume concentration of 60% to 80%, and the material-to-liquid ratio is 1:8 to 1:15 g / mL.

[0026] Yes, it's acceptable. The extraction methods are ultrasonic-assisted extraction or heating reflux extraction, with an extraction temperature of 50-70℃ and an extraction time of 2-3 hours.

[0027] The active ingredients in the *Mimosa pudica* extract described in this invention were qualitatively detected by ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS / MS) after redissolving an appropriate amount of the *Mimosa pudica* extract and comparing the chromatogram with that of a standard. The results confirmed the presence of 2,4-di-tert-butylphenol, 5-hydroxytryptamine, and mimosine. These three substances are characteristic marker active ingredients in the extract. This invention primarily focuses on qualitative research (i.e., the presence of these three components) and does not impose specific content limitations. The 15%–20% figure in the formulation refers to 15–20 grams of the total extract directly added when preparing 100 mL (or 100 g) of the final "soluble liquid".

[0028] The present invention relates to a mimosa extract formulation, wherein the formulation is a mimosa extract plant growth stimulant.

[0029] In the embodiments of this disclosure, the content of mimosa extract in the mimosa extract preparation is 15% to 20%.

[0030] In the embodiments of this disclosure, the mimosa extract plant growth stimulant comprises 15% to 20% mimosa extract, 3% to 6% dimethyl sulfoxide, 7% to 14% anhydrous ethanol, 10% NP-10, and water to make up to 100%.

[0031] The method for preparing the plant growth stimulant from the mimosa extract of the present invention includes: ① After crushing the mimosa plant, it was extracted with industrial ethanol at room temperature using ultrasound. The extract was then concentrated under reduced pressure using a rotary thin-film evaporator to obtain the total extract for later use. ② Preparation of plant growth stimulants from Mimosa extract: Prepare a soluble liquid using the Mimosa extract obtained in step ① according to the formula amount.

[0032] The present invention relates to the application of the mimosa extract plant growth stimulant in the preparation of crop fertilizers.

[0033] In embodiments of this disclosure, the crops include chili peppers and tomatoes, and the crop fertilizer is used to promote crop growth, increase yield, improve quality, and enhance stress resistance.

[0034] The mimosa plants in the following examples are from Northwest A&F University or purchased from Guangdong Kangmei Pharmaceutical Co., Ltd.

[0035] Example 1: Preparation of plant growth stimulants from Mimosa pudica extract: 10 kg of Mimosa pudica was pulverized and subjected to three ultrasonic extractions with industrial ethanol (material-to-liquid ratio 1:10) at 60℃, with each extraction lasting 1 hour (ultrasonic power 500 W). The extract was concentrated under reduced pressure using a rotary thin-film evaporator to obtain 696.43 g of total extract. A portion of the extract was sealed and stored at -4℃ for later use.

[0036] The formula for the plant growth stimulant extracted from Mimosa pudica was obtained through performance testing of the finished product, as detailed in Table 1.

[0037] Table 1. Optimal formulation of plant growth stimulants from Mimosa pudica extract.

[0038] Example 2: Seed germination test of active ingredients from Mimosa pudica extract. The specific implementation process and method are as follows: The viability of *Mimosa pudica* extract on tomato seeds was verified using an in-vessel seed germination assay. Healthy, plump seeds were selected, disinfected with 2.5% NaClO solution for 1 min, washed 2-3 times with distilled water, then disinfected with 75% ethanol solution for 30 s, washed 2-3 times with distilled water, and dried with filter paper. A layer of filter paper was placed in a glass dish (9 cm in diameter). Treatment groups were treated with 3.0 mL of *Mimosa pudica* extract plant growth stimulant at concentrations of 100 mg / L, 200 mg / L, and 400 mg / L, respectively, while the control group was treated with 3.0 mL of distilled water. Ten seeds were then dispersed on the filter paper, with each treatment repeated three times. The dishes were incubated in the dark at 25℃±1℃ for 14 days. Ten seeds were then selected, and the germination rate, plant height, and root length were recorded.

[0039] Table 2. Effects of active components of Mimosa pudica extract on tomato seed germination.

[0040] Note: This indicates a significant difference between the treatment and the control group. p <0.05).

[0041] The experimental results are shown in Table 2. 2,4-Di-tert-butylphenol and 5-hydroxytryptamine in the extract of Mimosa pudica have a promoting effect on the germination of tomato seeds and can accelerate seed germination and growth.

[0042] Example 3: Seed germination experiment using plant growth stimulants extracted from Mimosa pudica. The specific implementation process and methods are as follows: The in vitro seed germination test was used to verify the growth-promoting effect of Mimosa pudica extract plant growth stimulant on pepper and tomato seeds. Healthy and plump seeds were selected, disinfected with 2.5% NaClO solution for 1 min, washed 2-3 times with distilled water; then disinfected with 75% ethanol solution for 30 s, washed 2-3 times with distilled water, and dried with filter paper for later use. A layer of filter paper was placed in a glass dish (9 cm in diameter). The treatment groups were treated with 3.0 mL of Mimosa pudica extract plant growth stimulant at concentrations of 100 mg / L, 200 mg / L, and 400 mg / L, respectively. The control group was treated with 3.0 mL of distilled water. The positive control group was treated with 3.0 mL of 133 mg / L biostimulant Kelp seaweed extract (purchased from Yangling Agricultural Market, manufactured by Cape Company of South Africa). Ten seeds of the tested plant were then dispersed on the filter paper. Each treatment was repeated three times. The plants were cultured in the dark at 25℃±1℃ for 14 days. Ten seeds were selected, and the plant height and root length after germination were recorded.

[0043] Table 3-1 Effects of Mimosa pudica extract plant growth stimulants on chili seed germination

[0044] Note: This indicates a significant difference between the treatment and the control group. p <0.05).

[0045] Table 3-2 Effects of Mimosa pudica extract plant growth stimulants on tomato seed germination

[0046] Note: This indicates a significant difference between the treatment and the control group. p <0.05).

[0047] The experimental results are shown in Tables 3-1 and 3-2. The application of Mimosa extract plant growth stimulant has a promoting effect on the germination of tomato and pepper seeds, and can accelerate seed germination and growth.

[0048] Example 4: Screening the optimal concentration of plant growth stimulant from Mimosa pudica extract and applying the agent to pot experiments on tomatoes and peppers. The specific implementation process and methods are as follows: Tomato and pepper seedlings were transplanted into experimental pots, with one seedling per pot. The experiment began when the tomatoes had 4-6 true leaves and the peppers had 3-4 true leaves. The concentrations of the plant growth stimulant extracted from *Mimosa pudica* were set at 100 mg / L, 200 mg / L, and 400 mg / L, and applied via both spraying and root drenching, for a total of eight treatments. A foliar spray of the biostimulant Kelp algae extract (133 mg / L, optimal concentration) served as a positive control, while a water treatment served as a blank control. Each treatment was repeated 10 times. The experiment was conducted in a greenhouse at a temperature between 22℃ and 32℃ and a relative humidity between 50% and 90%. Spray once every 7 days for 3 consecutive times. The amount of liquid sprayed each time is 15 mL per plant. The spraying time is selected at 9:00 am. Random samples are taken at 7, 14 and 21 days after the first spraying to measure the stem diameter and plant height of plants under different treatments. The above-ground / underground fresh weight and dry weight of plants are measured at 21 days after the spraying.

[0049] Before each spraying, samples were taken. Five plants were randomly selected from each treatment. The diameter of the stem was measured using calipers, and the plant height was measured using a measuring tape.

[0050] Table 4-1 Effects of Mimosa extract plant growth stimulants spraying and drenching on tomato plant height

[0051] Table 4-2 Effects of Mimosa extract plant growth stimulants spraying and drenching on tomato stem diameter

[0052] Table 4-3 Effects of Mimosa pudica extract plant growth stimulants spraying and drenching on fresh and dry weight of tomatoes

[0053] Table 4-4 Effects of Mimosa pudica extract plant growth stimulants spraying and root irrigation on pepper plant height

[0054] Table 4-5 Effects of Mimosa extract plant growth stimulants spraying and root irrigation on pepper stem diameter

[0055] Table 4-6 Effects of Mimosa pudica extract plant growth stimulants spraying and root irrigation on the fresh and dry weight of chili peppers.

[0056] The experimental results are shown in Tables 4-1 to 4-6. Application of *Mimosa pudica* extract plant growth stimulant promoted the growth of both tomato and pepper plants. Spraying and root irrigation significantly increased plant height, stem diameter, fresh weight, and dry weight in both tomatoes and peppers. The optimal application method for *Mimosa pudica* extract plant growth stimulant in the pot experiment was spraying, and the optimal dosage for both tomato and pepper plants was 200 mg / L.

[0057] Example 5: The above-mentioned Mimosa soluble liquid was used in the production of tomatoes and peppers in a certain area of ​​Shaanxi Province. The specific implementation process and method are as follows: In the following experiment, the actual specific formula of the plant growth stimulant from the mimosa extract was as follows: 20% mimosa extract, 3% dimethyl sulfoxide, 7% anhydrous ethanol, 10% NP-10, and water to make up to 100%. The prepared solution was diluted to 200 mg / L, 400 mg / L and 600 mg / L for use.

[0058] The experiment was conducted at the Caoxinzhuang Farm experimental base in Yangling District, Shaanxi Province. Tomatoes and peppers were transplanted manually. Five treatments were set up (water control, positive control, 200 mg / L, 400 mg / L, and 600 mg / L), with three replicates for each treatment. A randomized block design was used, resulting in 30 plots, with two rows planted in each plot. Constant-volume spraying was performed using a backpack lever-type manual sprayer during the growth, flowering, and fruit-setting stages. Sampling was conducted on days 7, 14, and 21 after the third application.

[0059] Five plants were randomly selected for each treatment. The stem diameter was measured using calipers, and the plant height was measured using a measuring tape. The fresh weight of the tomato and pepper fruits was weighed directly, and the treatment was repeated three times.

[0060] Table 5-1 Effects of Mimosa pudica extract plant growth stimulants on pepper plant height

[0061] Note: This indicates a significant difference between the treatment and the control group. p <0.05).

[0062] Table 5-2 Effects of Mimosa pudica extract plant growth stimulants on pepper stem diameter

[0063] Note: This indicates a significant difference between the treatment and the control group. p <0.05).

[0064] Table 5-3 Effects of Mimosa pudica extract plant growth stimulants on fruit setting rate of pepper

[0065] Note: This indicates a significant difference between the treatment and the control group. p <0.05).

[0066] Table 5-4 Effects of Mimosa pudica extract plant growth stimulants on single fruit weight of chili peppers

[0067] Note: This indicates a significant difference between the treatment and the control group. p <0.05).

[0068] Table 5-5 Effects of Mimosa pudica extract plant growth stimulants on chili pepper yield

[0069] Table 5-6 Effects of Mimosa pudica extract plant growth stimulants on pepper fruit quality

[0070] Note: This indicates a significant difference between the treatment and the control group. p <0.05).

[0071] Table 5-7 Effects of Mimosa pudica extract plant growth stimulants on tomato plant height

[0072] Note: This indicates a significant difference between the treatment and the control group. p <0.05).

[0073] Table 5-8 Effects of Mimosa pudica extract plant growth stimulants on tomato stem diameter

[0074] Note: This indicates a significant difference between the treatment and the control group. p <0.05).

[0075] Table 5-9 Effects of Mimosa pudica extract plant growth stimulants on tomato fruit setting rate

[0076] Note: This indicates a significant difference between the treatment and the control group. p <0.05).

[0077] Table 5-10 Effects of Mimosa pudica extract plant growth stimulants on single fruit weight of tomato

[0078] Note: This indicates a significant difference between the treatment and the control group. p <0.05).

[0079] Table 5-11 Effects of Mimosa pudica extract plant growth stimulants on tomato yield

[0080] Table 5-12 Effects of Mimosa pudica extract plant growth stimulants on tomato fruit quality

[0081] Note: This indicates a significant difference between the treatment and the control group. p <0.05).

[0082] The test results are shown in Tables 5-1 to 5-12. Figures 1-4 As shown, the optimal application dose in field trials was 600 mg / L. Application of 600 mg / L of *Mimosa pudica* extract plant growth stimulant significantly improved the growth indicators and fruit quality of pepper and tomato plants. Plant height, stem diameter, and chlorophyll a and b contents of peppers and tomatoes were all significantly increased; fruit set rate, yield, vitamin C content, soluble sugar content, and organic acid content were all significantly increased. This indicates that 600 mg / L of *Mimosa pudica* extract plant growth stimulant has the effects of promoting growth, increasing yield, improving fruit quality, and enhancing plant stress resistance in peppers and tomatoes.

[0083] Finally, it should be noted that the above descriptions are merely specific embodiments of this application, enabling those skilled in the art to understand or implement this application. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of this application. Therefore, this application is not to be limited to the embodiments shown herein, but is to be accorded the widest scope consistent with the principles and novel features claimed herein.

Claims

1. Application of Mimosa pudica extract in the preparation of plant growth stimulants.

2. The application according to claim 1, characterized in that, The plants mentioned include chili peppers and tomatoes, and the plant growth stimulants are used to promote crop growth, increase yield, improve quality, and / or increase stress resistance.

3. The application according to claim 1 or 2, characterized in that, The mimosa extract contains 2,4-di-tert-butylphenol, 5-hydroxytryptamine, and mimosine.

4. The application according to claim 1 or 2, characterized in that, The method for preparing the mimosa extract includes: After drying and pulverizing the whole plant or aerial parts of Mimosa pudica, extract it using water, ethanol aqueous solution or methanol aqueous solution as extraction solvent at 40-80℃ for 1-4 hours. After filtration, collect the extract and concentrate it under reduced pressure to obtain Mimosa pudica extract.

5. The application according to claim 4, characterized in that, The extraction solvent is an aqueous ethanol solution with a volume concentration of 60% to 80%, and the material-to-liquid ratio is 1:8 to 1:15 g / mL.

6. The application according to claim 4, characterized in that, The extraction method is ultrasonic-assisted extraction or heating reflux extraction, with an extraction temperature of 50-70℃ and an extraction time of 2-3 hours.

7. A plant growth stimulant extracted from Mimosa pudica, characterized in that, The plant growth stimulant is formulated as a soluble liquid. The soluble liquid comprises 15%–20% mimosa extract, 3%–6% dimethyl sulfoxide, 7%–14% ethanol, 10% NP-10, and water to make up to 100%.

8. The method for preparing the plant growth stimulant from the mimosa extract according to claim 7, characterized in that, include: After drying and pulverizing the whole plant or aerial parts of Mimosa pudica, extracting it with water, ethanol aqueous solution or methanol aqueous solution as extraction solvent at 40-80℃ for 1-4 hours, collecting the extract after filtration, and concentrating it under reduced pressure to obtain Mimosa pudica extract. According to the formula, the obtained mimosa extract was prepared into a soluble liquid.

9. The method for preparing the plant growth stimulant from Mimosa pudica extract according to claim 8, characterized in that, The extraction solvent is an aqueous ethanol solution with a volume concentration of 60% to 80%, and the material-to-liquid ratio is 1:8 to 1:15 g / mL.

10. The method for preparing plant growth stimulant from Mimosa pudica extract according to claim 8, characterized in that, The extraction method is ultrasonic-assisted extraction or heating reflux extraction, with an extraction temperature of 50-70℃ and an extraction time of 2-3 hours.

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