A method for rapid planting and propagation using asexual propagation of perennial leguminous forage grasses

Abstract

This invention discloses a method for rapid propagation of perennial leguminous forage plants using asexual propagules, belonging to the field of plant propagation technology. The method includes the following steps: Step 1: Selecting mother plants of superior varieties, free from pests and diseases, with strong tillering ability, thick stems, and well-developed root systems, followed by fertilization, pruning, and pest control treatment; Step 2: Cutting all underground roots and lateral buds into 1cm long segments, then disinfecting and drying to obtain propagules; Step 3: Deeply tilling the soil, removing weed roots, mixing in well-rotted organic fertilizer, and adding modified fertilizer and modified antibacterial agents to obtain pretreated soil; Step 4: Planting the propagules, irrigating, fertilizing, weeding, and harvesting. This method uses asexual propagules of perennial leguminous forage plants as propagation materials instead of seeds, effectively improving the survival rate, tiller number, and annual forage yield of leguminous forage plants.

Description

Technical Field

[0001] This invention belongs to the field of plant cultivation and propagation technology, specifically relating to a method for rapid cultivation and propagation using asexual propagation of perennial leguminous forage grasses. Background Technology

[0002] Perennial leguminous forage grasses, as important ecological and livestock resources, possess significant advantages such as strong nitrogen fixation capacity, excellent palatability, high nutritional value, and soil structure improvement, playing a crucial role in grassland ecological restoration and sustainable livestock development. However, traditional sowing and propagation methods are limited by low seed germination rates, high risks of genetic variation, and long growth cycles (usually requiring 2-3 years to establish turf), making it difficult to meet the needs of large-scale planting. For example, while high-quality leguminous forage grasses such as alfalfa and red clover have great potential in the restoration of degraded grasslands, their seed-raising costs are high, and seedlings have weak resistance, resulting in low propagation efficiency. Furthermore, against the backdrop of global climate change, frequent droughts and pests and diseases further exacerbate the uncertainty of forage grass planting, urgently requiring innovative propagation technologies to overcome these bottlenecks. Summary of the Invention

[0003] The purpose of this invention is to provide a method for rapid planting and propagation of perennial leguminous forage using asexual propagules, in order to solve the technical problems of poor survival rate, tiller number and annual forage yield of leguminous forage in the prior art.

[0004] To achieve the above objectives, the present invention adopts the following technical solution:

[0005] This invention provides a method for rapid planting and propagation using asexual propagation organisms of perennial leguminous forage grasses, comprising the following steps:

[0006] Step 1: Select mother plants of superior variety, free from pests and diseases, with strong tillering ability, thick stems and well-developed root systems, and then carry out fertilization, pruning and pest control treatment;

[0007] Step 2: Cut all underground roots and lateral buds into 1cm long sections, then disinfect and dry them to obtain the propagation body;

[0008] Step 3: Deeply till the soil, remove weed roots, mix in well-rotted organic fertilizer, add modified fertilizer and modified antibacterial agent to obtain pretreated soil;

[0009] Step 4: Plant the propagules, irrigate, fertilize, weed, and harvest.

[0010] Preferably, in step one, a compound fertilizer with a nitrogen:phosphorus:potassium ratio of 10:5:5 is applied at a rate of 50-80 g / m³. 2 Prune away old and weak branches, retaining 4-6 strong lateral branches, and spray the leaves with an insecticide at a concentration of 0.3vt%-0.5vt%.

[0011] Preferably, in step three, the soil is deeply tilled to a depth of 25-35cm, the amount of well-rotted organic fertilizer added is 2000-3000 kg / mu, the amount of modified fertilizer added is 40-50 kg / mu, and the amount of modified antibacterial agent added is 2-2.5 kg / mu.

[0012] Preferably, the method for preparing the modified fertilizer includes the following steps:

[0013] Q1: Add m-fluorobenzonitrile, m-hydroxybenzoic acid and anhydrous potassium carbonate to a container, then add N,N-dimethylformamide, stir and heat to react. After the reaction is complete, filter, extract, wash, separate, wash the organic phase, dry, let stand, filter under vacuum and distill under reduced pressure to obtain compound 1.

[0014] Q2: Compound 1, sodium hydroxide, distilled water and anhydrous ethanol were added to a container in sequence, heated to reflux and stirred to react. After the reaction was completed, the mixture was distilled under reduced pressure, separated, the organic phase was washed, dried, allowed to stand, filtered, and distilled under reduced pressure to obtain compound 2.

[0015] Q3: Compound 2, N,N'-dicyclohexylcarbodiimide and N,N-dimethylformyl were slowly added to a container. Triethylamine was slowly added dropwise under ice bath conditions. The reaction was continued with stirring. Then dehydroabimethamine was added and the reaction was stirred at room temperature. After the reaction was completed, the mixture was diluted, extracted, washed, dried, rotary evaporated and purified to obtain compound 3.

[0016] Q4: Wash, crush, and dry the wheat straw, then soak it in ethanol, filter, and dry it to obtain pretreated wheat straw. Place it in distilled water containing compound 3, shake, filter, treat at high temperature, wash, and dry to obtain modified fertilizer.

[0017] In the above process, m-fluorobenzonitrile undergoes a nucleophilic substitution reaction with m-hydroxybenzoic acid to prepare compound 1. Subsequently, the nitrile group is first hydrolyzed to an amide, and then further hydrolyzed to a carboxylate group to obtain compound 2. Then, dehydroabimethamine reacts with compound 2 in the presence of N,N'-dicyclohexylcarbodiimide to form an amide bond, resulting in compound 3. Then, pretreated wheat straw is reacted with compound 3 under high temperature conditions, and the two undergo a hydrogenation reaction to prepare modified fertilizer.

[0018] Preferably, in Q1, the ratio of m-fluorobenzonitrile, m-hydroxybenzoic acid, anhydrous potassium carbonate, and N,N-dimethylformamide is (0.1-0.15) g : (0.09-0.13) g : (0.35-0.48) g : (1-2) mL, the reaction temperature is 90-110℃, the reaction time is 10-12 h, extraction is performed with ethyl acetate and distilled water, and the organic phase is washed with 5 wt% sodium hydroxide solution, distilled water, and saturated sodium chloride, respectively; in Q2, the ratio of compound 1, sodium hydroxide, distilled water, and anhydrous ethanol is (0.12-0.28) g : (0.22-0.35) g : (1.8-2.5) mL : (1.7-2.6) mL, the reaction temperature is 70-90℃, the reaction time is 8-10 h, and the organic phase is washed with 5 wt% sodium hydroxide solution, distilled water, and saturated sodium chloride, respectively.

[0019] Preferably, in Q3, the ratio of compound 2, N,N'-dicyclohexylcarbodiimide, N,N-dimethylformyl, triethylamine, and dehydroabimethamine is (0.32-0.43) g : (0.71-0.84) g : (3-4.5) mL : (0.28-0.35) mL : (0.21-0.35) g, the stirring reaction time is 30-45 min, and the stirring reaction time at room temperature is 15-20 h; in Q4, the ratio of pretreated wheat straw, compound 3, and distilled water is (50-80) g : (5-11) g : (100-200) mL, the high-temperature treatment temperature is 90-110℃, and the treatment time is 8-12 h.

[0020] Preferably, the preparation method of the modified antibacterial agent includes the following steps:

[0021] S1: 2,4-Dichloropyridine, 3-amino-2-thiophenecarboxylic acid and cesium carbonate were added to dimethyl sulfoxide and reacted under high temperature with stirring. Then ammonium chloride was added, and the mixture was extracted, washed, dried, vacuum treated and purified to obtain intermediate A.

[0022] S2: Add intermediate A to a container containing N,N-dimethylformamide, add sodium hydride while stirring in an ice bath, then add iodomethane and continue stirring, wash the organic layer, dry, vacuum treat, and purify to obtain the modified antibacterial agent.

[0023] The synthesis reaction formula for the modified antibacterial agent in the above process is as follows:

[0024]

[0025] The mass spectrometry analysis results of intermediate A were: m / z: 253.99 (100.0%), 255.99 (36.7%), 255.00 (11.0%), 256.99 (4.5%), 254.99 (1.5%), 257.98 (1.4%), 256.00 (1.0%); the mass spectrometry analysis results of the modified antibacterial agent were: m / z: 268.01 (100.0%), 270.00 (36.5%), 269.01 (12.9%), 271.01 (4.5%), 272.00 (1.5%), 270.01 (1.2%).

[0026] Preferably, in S1, the ratio of 2,4-dichloropyridine, 3-amino-2-thiophenecarboxylic acid, cesium carbonate, dimethyl sulfoxide, and ammonium chloride is (0.45-0.52) g : (0.58-0.67) g : (0.61-0.67) g : (5-7.5) mL : (20-30) mL, the high-temperature stirring reaction temperature is 100-110℃, and the stirring time is 20-24 h.

[0027] Preferably, in step S2, the ratio of intermediate A, N,N-dimethylformamide, sodium hydride, and iodomethane is (0.25-0.33) g : (3-5) mL : (0.002-0.005) g : (0.004-0.006) g. After adding sodium hydride for 10-15 min, iodomethane is added, and stirring is continued for 10-12 h. The mixture is then washed with saturated sodium chloride solution and dried with anhydrous sodium sulfate.

[0028] As a preferred option, in step four, during the planting process, the row spacing is 40-45cm, the plant spacing is 30-32.5cm, the number of plants is 5500-6000 per mu, the soil is covered to 1-2cm above the buds, and during irrigation, drip irrigation is carried out 2-3 times per week.

[0029] In summary, due to the adoption of the above technical solution, the beneficial effects of the present invention are:

[0030] This invention first prepares a modified fertilizer using m-fluorobenzonitrile, m-hydroxybenzoic acid, sodium hydroxide, N,N'-dicyclohexylcarbodiimide, dehydroabimethanil, and wheat straw as raw materials. Then, a modified antibacterial agent is prepared using 2,4-dichloropyridine, 3-amino-2-thiophenecarboxylic acid, sodium hydride, and iodomethane as raw materials. The modified fertilizer and modified antibacterial agent are added to a rapid propagation method for asexual reproduction of perennial leguminous forage plants. Simultaneously, using the asexual reproductions of perennial leguminous forage plants as propagation bodies instead of seeds for planting can effectively improve the survival rate, tiller number, and annual forage yield of the propagation bodies. The cross-linked structure contained in the modified fertilizer can optimize the soil microenvironment, and the pyridine ring contained in the modified antibacterial agent can act as a precursor for plant growth regulators, promoting adventitious root development and thus physiologically regulating the propagation bodies. Detailed Implementation

[0031] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only part of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts are within the scope of protection of the present invention.

[0032] Example 1: This example discloses a method for preparing a modified fertilizer, including the following steps:

[0033] Q1: 0.12 g m-fluorobenzonitrile, 0.11 g m-hydroxybenzoic acid and 0.42 g anhydrous potassium carbonate were added to a container, and then 1.5 mL N,N-dimethylformamide was added. The mixture was stirred and heated to 100 °C for 12 h. After the reaction was completed, the mixture was filtered, extracted with ethyl acetate and distilled water, washed, separated, and the organic phase was washed with 5 wt% sodium hydroxide solution, distilled water and saturated sodium chloride, respectively. The mixture was dried, allowed to stand, filtered under vacuum and distilled under reduced pressure to obtain compound 1.

[0034] Q2: 0.2g of compound 1, 0.28g of sodium hydroxide, 2.2mL of distilled water and 2.1mL of anhydrous ethanol were added to a container in sequence, and the mixture was heated to reflux at 80℃ and stirred for 10h. After the reaction was completed, the mixture was distilled under reduced pressure, separated, and the organic phase was washed with 5wt% sodium hydroxide solution, distilled water and saturated sodium chloride, respectively. The mixture was dried, allowed to stand, filtered, and distilled under reduced pressure to obtain compound 2.

[0035] Q3: 0.36 g of compound 2, 0.76 g of N,N'-dicyclohexylcarbodiimide and 3.75 mL of N,N-dimethylformyl were slowly added to a container. Under ice bath conditions, 0.31 mL of triethylamine was slowly added dropwise, and the reaction was stirred for 45 min. Then, 0.28 g of dehydroabimethamine was added, and the reaction was stirred at room temperature for 18 h. After the reaction was completed, the mixture was diluted, extracted, washed, dried, rotary evaporated, and purified to obtain compound 3.

[0036] Q4: Wash, crush, and dry the wheat straw, then soak it in ethanol, filter, and dry it to obtain pretreated wheat straw. Place 65g of pretreated wheat straw in 150mL of distilled water containing 8g of compound 3, shake, filter, treat at 100℃ for 12h, wash, and dry to obtain modified fertilizer.

[0037] This embodiment discloses a method for preparing a modified antibacterial agent, including the following steps:

[0038] S1: 0.48 g of 2,4-dichloropyridine, 0.62 g of 3-amino-2-thiophenecarboxylic acid and 0.64 g of cesium carbonate were added to 6.25 mL of dimethyl sulfoxide and stirred at 100 °C for 24 h. Then, 25 mL of ammonium chloride solution was added, and the mixture was extracted, washed, dried, vacuum treated and purified to obtain intermediate A.

[0039] S2: Add 0.29 g of intermediate A to a container containing 4 mL of N,N-dimethylformamide. Add 0.003 g of sodium hydride while stirring in an ice bath. After 10 min, add 0.005 g of iodomethane and continue stirring for 12 h. Wash the organic layer with saturated sodium chloride solution, dry with anhydrous sodium sulfate, vacuum treat, and purify to obtain the modified antibacterial agent.

[0040] This embodiment discloses a method for rapid planting and propagation using asexual propagation organisms of perennial leguminous forage grasses, including the following steps:

[0041] Step 1: Select mother plants of superior variety, free from pests and diseases, with strong tillering ability, thick stems, and well-developed root systems. Then fertilize (apply compound fertilizer with nitrogen:phosphorus:potassium = 10:5:5 at a rate of 60g / m²). 2 Prune away old and weak branches, retaining 4-6 strong lateral branches, and treat with insecticide (foliar spray with an insecticide at a concentration of 0.5 VT).

[0042] Step 2: Cut all underground roots and lateral buds into 1cm long sections, then disinfect and dry them to obtain the propagation body;

[0043] Step 3: Deeply till the soil to a depth of 30cm, remove weed roots, mix in well-rotted organic fertilizer (2500kg / mu), add modified fertilizer (45kg / mu) and modified antibacterial agent (2kg / mu) to obtain pretreated soil;

[0044] Step 4: Plant the propagules with a row spacing of 42.5cm and a plant spacing of 30cm, at a density of 5500-6000 plants per acre. Cover the soil with soil to 1-2cm above the buds, irrigate, drip irrigate 2-3 times per week, apply fertilizer, weed, and harvest.

[0045] Example 2: This example discloses a method for preparing a modified fertilizer, including the following steps:

[0046] Q1: 0.1 g m-fluorobenzonitrile, 0.09 g m-hydroxybenzoic acid and 0.35 g anhydrous potassium carbonate were added to a container, followed by 1 mL N,N-dimethylformamide. The mixture was stirred and heated to 100 °C for 12 h. After the reaction was completed, the mixture was filtered, extracted with ethyl acetate and distilled water, washed, separated, and the organic phase was washed with 5 wt% sodium hydroxide solution, distilled water and saturated sodium chloride, respectively. The mixture was dried, allowed to stand, filtered under vacuum and distilled under reduced pressure to obtain compound 1.

[0047] Q2: 0.12 g of compound 1, 0.22 g of sodium hydroxide, 1.8 mL of distilled water and 1.7 mL of anhydrous ethanol were added to a container in sequence, and the mixture was heated to reflux at 80 °C and stirred for 10 h. After the reaction was completed, the mixture was distilled under reduced pressure, separated, and the organic phase was washed with 5 wt% sodium hydroxide solution, distilled water and saturated sodium chloride, respectively. The mixture was dried, allowed to stand, filtered, and distilled under reduced pressure to obtain compound 2.

[0048] Q3: 0.32 g of compound 2, 0.71 g of N,N'-dicyclohexylcarbodiimide and 3 mL of N,N-dimethylformyl were slowly added to a container. Under ice bath conditions, 0.28 mL of triethylamine was slowly added dropwise, and the reaction was stirred for 45 min. Then, 0.21 g of dehydroabimethamine was added, and the reaction was stirred at room temperature for 18 h. After the reaction was completed, the mixture was diluted, extracted, washed, dried, rotary evaporated, and purified to obtain compound 3.

[0049] Q4: Wash, crush, and dry the wheat straw, then soak it in ethanol, filter, and dry it to obtain pretreated wheat straw. Place 50g of pretreated wheat straw in 100mL of distilled water containing 5g of compound 3, shake, filter, treat at 100℃ for 12h, wash, and dry to obtain modified fertilizer.

[0050] This embodiment discloses a method for preparing a modified antibacterial agent, including the following steps:

[0051] S1: 0.45 g of 2,4-dichloropyridine, 0.58 g of 3-amino-2-thiophenecarboxylic acid and 0.61 g of cesium carbonate were added to 5 mL of dimethyl sulfoxide and stirred at 100 °C for 24 h. Then, 20 mL of ammonium chloride solution was added, and the mixture was extracted, washed, dried, vacuum treated and purified to obtain intermediate A.

[0052] S2: Add 0.25g of intermediate A to a container containing 3mL of N,N-dimethylformamide. Add 0.002g of sodium hydride while stirring in an ice bath. After 10min, add 0.004g of iodomethane and continue stirring for 12h. Wash the organic layer with saturated sodium chloride solution, dry with anhydrous sodium sulfate, vacuum treat, and purify to obtain the modified antibacterial agent.

[0053] This embodiment discloses a method for rapid planting and propagation using asexual propagation organisms of perennial leguminous forage grasses, including the following steps:

[0054] Step 1: Select mother plants of superior variety, free from pests and diseases, with strong tillering ability, thick stems, and well-developed root systems. Then fertilize (apply compound fertilizer with nitrogen:phosphorus:potassium = 10:5:5 at a rate of 60g / m²). 2 Prune away old and weak branches, retaining 4-6 strong lateral branches, and treat with insecticide (foliar spray with an insecticide at a concentration of 0.5 VT).

[0055] Step 2: Cut all underground roots and lateral buds into 1cm long sections, then disinfect and dry them to obtain the propagation body;

[0056] Step 3: Deeply till the soil to a depth of 30cm, remove weed roots, mix in well-rotted organic fertilizer (2500kg / mu), add modified fertilizer (45kg / mu) and modified antibacterial agent (2kg / mu) to obtain pretreated soil;

[0057] Step 4: Plant the propagules with a row spacing of 42.5cm and a plant spacing of 30cm, at a density of 5500-6000 plants per acre. Cover the soil with soil to 1-2cm above the buds, irrigate, drip irrigate 2-3 times per week, apply fertilizer, weed, and harvest.

[0058] Example 3: This example discloses a method for preparing a modified fertilizer, including the following steps:

[0059] Q1: 0.15 g m-fluorobenzonitrile, 0.13 g m-hydroxybenzoic acid and 0.48 g anhydrous potassium carbonate were added to a container, followed by 2 mL N,N-dimethylformamide. The mixture was stirred and heated to 100 °C for 12 h. After the reaction was completed, the mixture was filtered, extracted with ethyl acetate and distilled water, washed, separated, and the organic phase was washed with 5 wt% sodium hydroxide solution, distilled water and saturated sodium chloride, respectively. The mixture was dried, allowed to stand, filtered under vacuum and distilled under reduced pressure to obtain compound 1.

[0060] Q2: 0.28 g of compound 1, 0.35 g of sodium hydroxide, 2.5 mL of distilled water and 2.6 mL of anhydrous ethanol were added to a container in sequence, and the mixture was heated to reflux at 80 °C and stirred for 10 h. After the reaction was completed, the mixture was distilled under reduced pressure, separated, and the organic phase was washed with 5 wt% sodium hydroxide solution, distilled water and saturated sodium chloride, respectively. The mixture was dried, allowed to stand, filtered, and distilled under reduced pressure to obtain compound 2.

[0061] Q3: 0.43 g of compound 2, 0.84 g of N,N'-dicyclohexylcarbodiimide and 4.5 mL of N,N-dimethylformyl were slowly added to a container. Under ice bath conditions, 0.35 mL of triethylamine was slowly added dropwise, and the reaction was stirred for 45 min. Then, 0.35 g of dehydroabimethamine was added, and the reaction was stirred at room temperature for 18 h. After the reaction was completed, the mixture was diluted, extracted, washed, dried, rotary evaporated, and purified to obtain compound 3.

[0062] Q4: Wash, crush, and dry the wheat straw, then soak it in ethanol, filter, and dry it to obtain pretreated wheat straw. Place 80g of the pretreated wheat straw in 200mL of distilled water containing 11g of compound 3, shake, filter, treat at 100℃ for 12h, wash, and dry to obtain modified fertilizer.

[0063] This embodiment discloses a method for preparing a modified antibacterial agent, including the following steps:

[0064] S1: 0.52 g of 2,4-dichloropyridine, 0.67 g of 3-amino-2-thiophenecarboxylic acid and 0.67 g of cesium carbonate were added to 7.5 mL of dimethyl sulfoxide and stirred at 100 °C for 24 h. Then, 30 mL of ammonium chloride solution was added, and the mixture was extracted, washed, dried, vacuum treated and purified to obtain intermediate A.

[0065] S2: Add 0.33g of intermediate A to a container containing 5mL of N,N-dimethylformamide. Add 0.005g of sodium hydride while stirring in an ice bath. After 10min, add 0.006g of iodomethane and continue stirring for 12h. Wash the organic layer with saturated sodium chloride solution, dry with anhydrous sodium sulfate, vacuum treat, and purify to obtain the modified antibacterial agent.

[0066] This embodiment discloses a method for rapid planting and propagation using asexual propagation organisms of perennial leguminous forage grasses, including the following steps:

[0067] Step 1: Select mother plants of superior variety, free from pests and diseases, with strong tillering ability, thick stems, and well-developed root systems. Then fertilize (apply compound fertilizer with nitrogen:phosphorus:potassium = 10:5:5 at a rate of 60g / m²). 2 Prune away old and weak branches, retaining 4-6 strong lateral branches, and treat with insecticide (foliar spray with an insecticide at a concentration of 0.5 VT).

[0068] Step 2: Cut all underground roots and lateral buds into 1cm long sections, then disinfect and dry them to obtain the propagation body;

[0069] Step 3: Deeply till the soil to a depth of 30cm, remove weed roots, mix in well-rotted organic fertilizer (2500kg / mu), add modified fertilizer (45kg / mu) and modified antibacterial agent (2kg / mu) to obtain pretreated soil;

[0070] Step 4: Plant the propagules with a row spacing of 42.5cm and a plant spacing of 30cm, at a density of 5500-6000 plants per acre. Cover the soil with soil to 1-2cm above the buds, irrigate, drip irrigate 2-3 times per week, apply fertilizer, weed, and harvest.

[0071] Example 4: This example discloses a method for preparing a modified fertilizer, including the following steps:

[0072] Q1: 0.11 g m-fluorobenzonitrile, 0.1 g m-hydroxybenzoic acid and 0.39 g anhydrous potassium carbonate were added to a container, followed by 1.2 mL N,N-dimethylformamide. The mixture was stirred and heated to 100 °C for 12 h. After the reaction was completed, the mixture was filtered, extracted with ethyl acetate and distilled water, washed, separated, and the organic phase was washed with 5 wt% sodium hydroxide solution, distilled water and saturated sodium chloride, respectively. The mixture was dried, allowed to stand, filtered under vacuum and distilled under reduced pressure to obtain compound 1.

[0073] Q2: 0.16 g of compound 1, 0.26 g of sodium hydroxide, 1.9 mL of distilled water and 1.9 mL of anhydrous ethanol were added to a container in sequence, and the mixture was heated to reflux at 80 °C and stirred for 10 h. After the reaction was completed, the mixture was distilled under reduced pressure, separated, and the organic phase was washed with 5 wt% sodium hydroxide solution, distilled water and saturated sodium chloride, respectively. The mixture was dried, allowed to stand, filtered, and distilled under reduced pressure to obtain compound 2.

[0074] Q3: 0.34 g of compound 2, 0.73 g of N,N'-dicyclohexylcarbodiimide and 3.2 mL of N,N-dimethylformyl were slowly added to a container. Under ice bath conditions, 0.29 mL of triethylamine was slowly added dropwise, and the reaction was stirred for 45 min. Then, 0.24 g of dehydroabimethamine was added, and the reaction was stirred at room temperature for 18 h. After the reaction was completed, the mixture was diluted, extracted, washed, dried, rotary evaporated, and purified to obtain compound 3.

[0075] Q4: Wash, crush, and dry the wheat straw, then soak it in ethanol, filter, and dry it to obtain pretreated wheat straw. Place 60g of pretreated wheat straw in 110mL of distilled water containing 7g of compound 3, shake, filter, treat at 100℃ for 12h, wash, and dry to obtain modified fertilizer.

[0076] This embodiment discloses a method for preparing a modified antibacterial agent, including the following steps:

[0077] S1: 0.47 g of 2,4-dichloropyridine, 0.6 g of 3-amino-2-thiophenecarboxylic acid and 0.62 g of cesium carbonate were added to 5.5 mL of dimethyl sulfoxide and stirred at 100 °C for 24 h. Then, 22 mL of ammonium chloride solution was added, and the mixture was extracted, washed, dried, vacuum treated and purified to obtain intermediate A.

[0078] S2: Add 0.27g of intermediate A to a container containing 3.5mL of N,N-dimethylformamide. Add 0.004g of sodium hydride while stirring in an ice bath. After 10min, add 0.005g of iodomethane and continue stirring for 12h. Wash the organic layer with saturated sodium chloride solution, dry with anhydrous sodium sulfate, vacuum treat, and purify to obtain the modified antibacterial agent.

[0079] This embodiment discloses a method for rapid planting and propagation using asexual propagation organisms of perennial leguminous forage grasses, including the following steps:

[0080] Step 1: Select mother plants of superior variety, free from pests and diseases, with strong tillering ability, thick stems, and well-developed root systems. Then fertilize (apply compound fertilizer with nitrogen:phosphorus:potassium = 10:5:5 at a rate of 60g / m²). 2 Prune away old and weak branches, retaining 4-6 strong lateral branches, and treat with insecticide (foliar spray with an insecticide at a concentration of 0.5 VT).

[0081] Step 2: Cut all underground roots and lateral buds into 1cm long sections, then disinfect and dry them to obtain the propagation body;

[0082] Step 3: Deeply till the soil to a depth of 30cm, remove weed roots, mix in well-rotted organic fertilizer (2500kg / mu), add modified fertilizer (45kg / mu) and modified antibacterial agent (2kg / mu) to obtain pretreated soil;

[0083] Step 4: Plant the propagules with a row spacing of 42.5cm and a plant spacing of 30cm, at a density of 5500-6000 plants per acre. Cover the soil with soil to 1-2cm above the buds, irrigate, drip irrigate 2-3 times per week, apply fertilizer, weed, and harvest.

[0084] Comparative Example 1: Compared with Example 1, Comparative Example 1 did not add modified fertilizer during the implementation of the planting and propagation method, and all other conditions remained unchanged.

[0085] Comparative Example 2: Compared with Example 1, Comparative Example 2 did not add modified antibacterial agents during the implementation of the planting and propagation method, and all other conditions remained unchanged.

[0086] Comparative Example 3: Compared with Example 1, Comparative Example 3 used seeds directly for propagation during the propagation process, and did not add modified fertilizer or modified antibacterial agent, while keeping other conditions unchanged.

[0087] Experimental Example: Alfalfa was used as the experimental leguminous forage. Propagation was carried out using the methods described in Examples 1-4 and Comparative Examples 1-3. The survival rate of the propagules, the number of tillers, and the annual forage yield were tested. The test results are shown in Table 1.

[0088]

[0089] As shown in Table 1, the methods described in Examples 1-4 of this invention can effectively promote the planting and propagation of leguminous forage. A comparison between Comparative Example 1 and Examples 1-4 shows that applying modified fertilizer can effectively improve the survival rate of leguminous forage propagules, the number of tillers, and the annual forage yield. Similarly, a comparison between Comparative Example 2 and Examples 1-4 shows that applying modified fertilizer can effectively improve the survival rate of leguminous forage propagules, the number of tillers, and the annual forage yield.

[0090] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

[0091] The preferred embodiments of the present invention disclosed above are merely illustrative of the invention. These preferred embodiments do not exhaustively describe all details, nor do they limit the invention to specific implementations. Clearly, many modifications and variations can be made based on the content of this specification. This specification selects and specifically describes these embodiments to better explain the principles and practical applications of the invention, thereby enabling those skilled in the art to better understand and utilize the invention. The invention is limited only by the claims and their full scope and equivalents.

Claims

1. A method for rapid planting and propagation using asexual propagation organisms of perennial leguminous forage grasses, characterized in that, Includes the following steps: Step 1: Select mother plants of superior variety, free from pests and diseases, with strong tillering ability, thick stems and well-developed root systems, and then carry out fertilization, pruning and pest control treatment; Step 2: Cut all underground roots and lateral buds into 1cm long sections, then disinfect and dry them to obtain the propagation body; Step 3: Deeply till the soil, remove weed roots, mix in well-rotted organic fertilizer, add modified fertilizer and modified antibacterial agent to obtain pretreated soil; Step 4: Plant the propagules, irrigate, fertilize, weed, and harvest; The method for preparing the modified fertilizer, Includes the following steps: Q1: Add m-fluorobenzonitrile, m-hydroxybenzoic acid and anhydrous potassium carbonate to a container, then add N,N-dimethylformamide, stir and heat to react. After the reaction is complete, filter, extract, wash, separate, wash the organic phase, dry, let stand, filter under vacuum and distill under reduced pressure to obtain compound 1. Q2: Compound 1, sodium hydroxide, distilled water and anhydrous ethanol were added to a container in sequence, heated to reflux and stirred to react. After the reaction was completed, the mixture was distilled under reduced pressure, separated, the organic phase was washed, dried, allowed to stand, filtered, and distilled under reduced pressure to obtain compound 2. Q3: Compound 2, N,N'-dicyclohexylcarbodiimide and N,N-dimethylformyl were slowly added to a container. Triethylamine was slowly added dropwise under ice bath conditions. The reaction was continued with stirring. Then dehydroabimethamine was added and the reaction was stirred at room temperature. After the reaction was completed, the mixture was diluted, extracted, washed, dried, rotary evaporated and purified to obtain compound 3. Q4: Wash, crush, and dry the wheat straw, then soak it in ethanol, filter, and dry it to obtain pretreated wheat straw. Place it in distilled water containing compound 3, shake, filter, treat at high temperature, wash, and dry to obtain modified fertilizer. The preparation method of the modified antibacterial agent includes the following steps: S1: 2,4-Dichloropyridine, 3-amino-2-thiophenecarboxylic acid and cesium carbonate were added to dimethyl sulfoxide and reacted under high temperature with stirring. Then ammonium chloride was added, and the mixture was extracted, washed, dried, vacuum treated and purified to obtain intermediate A. S2: Add intermediate A to a container containing N,N-dimethylformamide, add sodium hydride while stirring in an ice bath, then add iodomethane and continue stirring, wash the organic layer, dry, vacuum treat, purify, and obtain the modified antibacterial agent; The perennial leguminous forage grass mentioned is alfalfa.

2. The method for rapid planting and propagation of perennial leguminous forage grasses using asexual propagation organisms according to claim 1, characterized in that, In step one, a compound fertilizer with a nitrogen:phosphorus:potassium ratio of 10:5:5 is applied at a rate of 50-80 g / m³. 2 Prune away old and weak branches, retaining 4-6 strong lateral branches, and spray the leaves with an insecticide at a concentration of 0.3vt%-0.5vt%.

3. The method for rapid planting and propagation of perennial leguminous forage grasses using asexual propagation organisms according to claim 1, characterized in that, In step three, the soil is deeply tilled to a depth of 25-35cm, the amount of well-rotted organic fertilizer added is 2000-3000 kg / mu, the amount of modified fertilizer added is 40-50 kg / mu, and the amount of modified antibacterial agent added is 2-2.5 kg / mu.

4. The method for rapid planting and propagation of perennial leguminous forage grasses using asexual propagation organisms according to claim 1, characterized in that, In Q1, the ratio of m-fluorobenzonitrile, m-hydroxybenzoic acid, anhydrous potassium carbonate, and N,N-dimethylformamide is (0.1-0.15) g : (0.09-0.13) g : (0.35-0.48) g : (1-2) mL. The reaction temperature is 90-110℃, and the reaction time is 10-12 h. Extraction is performed with ethyl acetate and distilled water, and the organic phase is washed with 5 wt% sodium hydroxide solution, distilled water, and saturated sodium chloride, respectively. In Q2, the ratio of compound 1, sodium hydroxide, distilled water, and anhydrous ethanol is (0.12-0.28) g : (0.22-0.35) g : (1.8-2.5) mL : (1.7-2.6) mL. The reaction temperature is refluxed and stirred at 70-90℃, and the reaction time is 8-10 h. The organic phase is washed with 5 wt% sodium hydroxide solution, distilled water, and saturated sodium chloride, respectively.

5. The method for rapid planting and propagation of perennial leguminous forage grasses using asexual propagation organisms according to claim 1, characterized in that, In Q3, the ratio of compound 2, N,N'-dicyclohexylcarbodiimide, N,N-dimethylformyl, triethylamine, and dehydroabimethamine is (0.32-0.43) g : (0.71-0.84) g : (3-4.5) mL : (0.28-0.35) mL : (0.21-0.35) g, the stirring reaction time is 30-45 min, and the stirring reaction time at room temperature is 15-20 h. In Q4, the ratio of pretreated wheat straw, compound 3, and distilled water is (50-80) g : (5-11) g : (100-200) mL, the high temperature treatment temperature is 90-110℃, and the treatment time is 8-12 h.

6. The method for rapid planting and propagation of perennial leguminous forage grasses using asexual propagation organisms according to claim 1, characterized in that, In S1, the ratio of 2,4-dichloropyridine, 3-amino-2-thiophenecarboxylic acid, cesium carbonate, dimethyl sulfoxide, and ammonium chloride is (0.45-0.52) g : (0.58-0.67) g : (0.61-0.67) g : (5-7.5) mL : (20-30) mL. The high-temperature stirring reaction temperature is 100-110℃, and the stirring time is 20-24 h.

7. The method for rapid planting and propagation of perennial leguminous forage grasses using asexual propagation organisms according to claim 1, characterized in that, In S2, the ratio of intermediate A, N,N-dimethylformamide, sodium hydride and iodomethane is (0.25-0.33) g : (3-5) mL : (0.002-0.005) g : (0.004-0.006) g. After adding sodium hydride for 10-15 min, iodomethane is added, and stirring is continued for 10-12 h. The mixture is washed with saturated sodium chloride solution and dried with anhydrous sodium sulfate.

8. The method for rapid planting and propagation of perennial leguminous forage grasses using asexual propagation organisms according to claim 1, characterized in that, In step four, during the planting process, the row spacing is 40-45cm, the plant spacing is 30-32.5cm, and the number of plants is 5500-6000 per mu. The soil is covered to 1-2cm above the buds. During irrigation, drip irrigation is carried out 2-3 times per week.

Images