Underground propagule composite excitant for ecological restoration and preparation method of underground propagule composite excitant
By using a composite activator for underground propagules in ecological restoration, and by employing multi-dimensional regulation and signal simulation technology, the problem of slow germination of underground propagules in grassland restoration has been solved, enabling rapid grassland recovery and the formation of diverse communities, thereby improving grassland productivity.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-12-29
- Publication Date
- 2026-03-31
AI Technical Summary
Existing technologies suffer from problems such as dormancy, slow germination, and uneven germination of native grass species, resulting in poor grassland restoration effects. Reseeding techniques also face challenges such as low seed quality, difficulty in mechanized sowing, monotonous community composition, and poor stability.
The method employs a composite activator for underground propagation in ecological restoration, which includes a combination of gibberellin, 6-benzylaminopurine, and indolebutyric acid, along with jasmonic acid slow-release microspheres and composite microbial agents. Through multi-dimensional regulation, it promotes the germination and growth of underground propagation, simulates natural environmental signals, improves soil microecology, provides nutrients, and improves soil structure.
It significantly increases bud bank density and vigor, promotes rapid germination, forms a stable and diverse community, enhances grassland restoration effectiveness and efficiency, and increases grassland coverage and biodiversity.
Smart Images
Figure SMS_1
Abstract
Description
Technical Field
[0001] This application relates to the field of activator preparation technology, and in particular to a composite activator for underground reproductive bodies used in ecological restoration and its preparation method. Background Technology
[0002] Due to long-term unreasonable use of grasslands, coupled with frequent climate disasters and biological hazards such as rodents and insects, more than 60% of grasslands have experienced varying degrees of degradation. Both vegetation and soil conditions have changed, primarily manifested in: a reduction in the types and proportion of high-yield, high-quality forage grasses; the spread of toxic and harmful species that livestock do not graze on; and a decrease in soil organic matter content. These changes have led to a significant reduction in grassland productivity, severely restricting the grasslands' ecological and productive functions.
[0003] In grassland restoration, increasing the density and potential energy of underground propagules has become a key plant factor. Underground propagules include underground asexual propagation bud banks and soil seed banks, and their density and vitality directly affect the recovery speed and stability of grassland vegetation. Activating underground propagules and promoting their rapid germination and growth is a key technology for regulating grassland restoration capacity. However, the dormancy, slow germination, and uneven germination of native grass species severely affect the effectiveness of grassland restoration. The use of native species for reseeding and disturbance has advanced the theoretical and technical research of multi-species reseeding, but reseeding technology still faces many challenges in large-scale ecological restoration practices. Problems such as low seed quality and difficulty in mechanized sowing result in monotonous reseeded community composition, poor stability, and low seed germination and survival rates, thus limiting the promotion of reseeding technology.
[0004] Therefore, developing a plant underground propagule activator that can effectively activate underground propagules, promote the rapid germination and growth of compound reseeded seeds, and stimulate the native seed bank is of great significance for realizing the technology of multi-species compound reseeding to improve grasslands. Summary of the Invention
[0005] To address the problems of insufficient and weak propagules in existing degraded grasslands, this application provides a composite activator for underground propagules in ecological restoration and its preparation method.
[0006] This application provides a composite activator for underground reproductive organisms used in ecological restoration, employing the following technical solution: A composite activator for underground reproductive organisms used in ecological restoration comprises, by weight, 0.2-0.3 parts gibberellin, 0.2-0.5 parts 6-benzylaminopurine, 0.2-0.3 parts indolebutyric acid, 0.5-0.7 parts sodium nitroprusside, 0.4-0.5 parts hydrogen peroxide, 1-2 parts jasmonic acid sustained-release microspheres, 5-7 parts composite amino acids, 1-3 parts composite microbial agent sustained-release microspheres, 2-3 parts seaweed extract, 10-13 parts humic acid, 2-3 parts potassium dihydrogen phosphate, 60-74 parts ethanol, and 1000 parts water.
[0007] Preferably, the composite amino acids include proline, tryptophan, and valine; the mass ratio of proline, tryptophan, and valine is (2-3):(1-2):1.
[0008] Preferably, the preparation method of the composite microbial agent sustained-release microspheres includes the following steps: After mixing the composite microbial agent suspension and the composite wall material solution at a volume ratio of 1:(2-2.5), the mixture was incubated in a shaker at 32-36℃ and 180-200rpm for 1-2 hours. The resulting mixture was then injected into a 2% calcium chloride solution for cross-linking for 1.5-2.5 hours. After washing several times with sterile water, wet microspheres were obtained. The wet microspheres were then dried at 40-45℃ to obtain the composite microbial agent slow-release microspheres.
[0009] Preferably, the bacterial cell concentration of the composite microbial agent suspension is 5 × 10⁻⁶. 8 -10×10 8 CFU / mL; the compound microbial agent includes purple non-sulfur bacteria, Pseudomonas aeruginosa, and yeast; the volume ratio of the purple non-sulfur bacteria solution, Pseudomonas aeruginosa solution, and yeast solution is (2.5-3):(1-1.5):1.
[0010] Preferably, the composite wall material includes sodium alginate, starch, sodium carboxymethyl cellulose, and meat and bone meal; The preparation method of the composite wall material solution includes the following steps: Sodium alginate is added to water and stirred at 60-70℃ until dissolved to obtain a sodium alginate solution with a concentration of 2-4%. Add starch to water, stir well, then add boiling water and heat and stir until transparent to obtain a starch solution with a concentration of 1-2%. Add sodium carboxymethyl cellulose to water, let stand for 2-3 hours, and then stir until homogeneous to obtain a sodium carboxymethyl cellulose solution with a concentration of 0.5-1%. After grinding the meat and bone meal, sieve it, add it to water, sonicate it at 60-70℃ for 30-40 minutes, filter it to remove insoluble matter, and obtain a meat and bone meal solution with a concentration of 5-10%. Sodium alginate solution, starch solution, sodium carboxymethyl cellulose solution, and meat and bone meal solution are mixed in a volume ratio of (1.8-2):(1-1.2):(1-1.2):1 and stirred at 60-70℃ for 1-2 hours. The pH is then adjusted to 6.5-7.0 to obtain a composite wall material solution.
[0011] Preferably, the jasmonic acid sustained-release microspheres are prepared from the following raw materials in parts by weight: 10-15 parts jasmonic acid, 50-75 parts carboxymethyl porous starch, 5-7.5 parts sodium alginate, 200-300 parts water, 40-60 parts ethanol, 200-300 parts calcium chloride solution with a concentration of 2%, 240-360 parts corn oil, and 6-9 parts emulsifier.
[0012] Preferably, the carboxymethyl porous starch is prepared from the following raw materials in parts by weight: 10-15 parts porous starch, 100-150 parts ethanol solution, 16-24 parts sodium hydroxide, and 58-87 parts sodium chloroacetate. The porous starch is prepared from the following raw materials in parts by weight: 20-30 parts starch, 200-300 parts citrate-sodium dihydrogen phosphate buffer, and 2.8-4.2 parts compound enzyme; The complex enzyme comprises α-amylase and saccharifying enzyme in a mass ratio of 1:(1.5-1.7).
[0013] Preferably, the method for preparing the carboxymethyl porous starch includes the following steps: By weight, 10-15 parts of porous starch were added to 100-150 parts of ethanol solution and stirred until homogeneous to obtain a starch slurry solution. After preheating at 30-40℃ for 20-25 min, 12-18 parts of sodium hydroxide were added, and the alkalization reaction was carried out at 30-40℃ for 30-40 min. The temperature was then raised to 45-55℃, and 58-87 parts of sodium chloroacetate and 4-6 parts of sodium hydroxide were added. The etherification reaction was carried out at 45-55℃ for 3-5 h. Subsequently, the pH of the starch slurry solution was adjusted to 6.5-7.0 to end the etherification reaction. After centrifugation and washing, the product was dried at 40-50℃ overnight and sieved to obtain carboxymethyl porous starch. The method for preparing the porous starch includes the following steps: By weight, 20-30 parts of starch were added to 200-300 parts of citrate-sodium dihydrogen phosphate buffer solution and stirred to obtain a starch emulsion solution. The solution was preheated at 50-60℃ for 20-30 min, and then 2.8-4.2 parts of compound enzyme were added. The solution was reacted at 50-60℃ for 7-9 h. After the reaction was completed, the product was centrifuged, washed with distilled water, freeze-dried for 12-24 h, and sieved to obtain porous starch.
[0014] Preferably, the method for preparing the jasmonic acid sustained-release microspheres includes the following steps: Carboxymethyl porous starch and sodium alginate were added to water and stirred thoroughly until dissolved to obtain a composite wall material solution. Jasmonic acid was added to ethanol and magnetically stirred until dissolved to obtain a jasmonic acid solution. The composite wall material solution and the jasmonic acid solution were mixed and ultrasonically treated at 60-70℃ for 30-50 min to obtain a mixed solution. The emulsifier was stirred evenly in corn oil and then added to the mixed solution. After shearing and emulsification at 8000-10000 rpm for 10-20 min, a 2% calcium chloride solution was added for crosslinking reaction for 30-50 min. After centrifugation and washing, wet microspheres were obtained. After freeze-drying, jasmonic acid sustained-release microspheres were obtained.
[0015] This application provides a method for preparing a composite activator for underground reproductive organisms used in ecological restoration, which adopts the following technical solution: A method for preparing a composite activator for underground reproductive organisms used in ecological restoration includes the following steps: Sodium nitroprusside, potassium dihydrogen phosphate, hydrogen peroxide, and humic acid were added to water in sequence and stirred until completely dissolved to obtain mixed solution a. Gibberellin, 6-benzylaminopurine, and indolebutyric acid were added to ethanol and stirred until completely dissolved to obtain mixed solution b. After mixing solutions a and b thoroughly, while maintaining stirring, add seaweed extract, compound amino acids, jasmonic acid slow-release microspheres, and compound microbial agent slow-release microspheres in sequence. Continue stirring for 30-60 minutes to obtain the underground propagation compound activator for ecological restoration.
[0016] In summary, this application includes at least one of the following beneficial technical effects: 1. This application promotes the activation and germination of underground propagules through multi-dimensional regulation. The combination of gibberellin, 6-benzylaminopurine, and indolebutyric acid targets key aspects such as seed dormancy breaking, bud bank meristem activation, and root development, synergistically promoting the germination and growth of underground propagules. Sodium nitroprusside and hydrogen peroxide, as signaling molecules, mimic redox signals and nitrogen oxide signals in the natural environment, further breaking seed dormancy, initiating the germination process, and improving germination synchronicity. Jasmonic acid, as an important plant signaling molecule, is designed with slow-release microspheres for continuous release, enhancing plant stress resistance while avoiding potential damage to plants from a single high-concentration release. The compound microbial agent contains purple non-sulfur bacteria, Pseudomonas aeruginosa, and yeast. These strains have multiple functions such as nitrogen fixation, phosphorus solubilization, and growth promotion, which can improve the soil microecological environment and promote plant growth. Through slow-release microspheres, the microbial agent is slowly released into the soil, prolonging its activity maintenance time and improving the stability and effectiveness of the agent. The compound amino acids and potassium dihydrogen phosphate provide the nutrients needed for plant growth, promoting cell division, elongation, and protein synthesis, thereby enhancing plant vigor and stress resistance. Seaweed extract and humic acid are rich in various bioactive substances and organic matter, which can improve soil structure, enhance soil water retention and nutrient retention capacity, and stimulate plant root growth. In summary, the compound activator provided in this application can not only significantly increase bud bank density and vigor, break seed dormancy, and promote rapid germination, but also form a stable and diverse community, significantly improving the effectiveness and efficiency of grassland restoration. It can be widely applied to the vegetation restoration of degraded grasslands, increasing grassland cover and biodiversity. Detailed Implementation
[0017] The present application will be further described in detail below with reference to the embodiments.
[0018] The chemical reagents used in the preparation examples, embodiments, and comparative examples provided in this invention are all commercially available products.
[0019] Preparation Example 1: Preparation of Composite Microbial Agent Slow-Release Microspheres Preparation Example 1.1 S1. Add sodium alginate to deionized water and stir at 60°C until dissolved to obtain a 2% sodium alginate solution; Add potato starch to deionized water, stir well, then add boiling water and heat and stir until transparent to obtain a starch solution with a concentration of 1%. Sodium carboxymethyl cellulose was added to deionized water, allowed to stand for 2 hours, and then stirred until homogeneous to obtain a sodium carboxymethyl cellulose solution with a concentration of 0.5%. After grinding the meat and bone meal into powder, it was passed through a 100-mesh sieve, added to deionized water, and ultrasonically treated at 60°C for 30 minutes. The insoluble matter was then removed by filtration to obtain a 5% meat and bone meal solution. Sodium alginate solution, starch solution, sodium carboxymethyl cellulose solution, and meat and bone meal solution were mixed in a volume ratio of 1.8:1:1:1 and stirred at 60°C for 1 hour. The pH was then adjusted to 6.5 with 1M sodium hydroxide solution to obtain the composite wall material solution. S2. The composite microbial agent suspension and the composite wall material solution were mixed evenly at a volume ratio of 1:2; the mixture was then shaken and cultured at 32℃ and 180 rpm for 1 h; the resulting mixed solution was then injected into 200 mL of 2% calcium chloride solution for cross-linking for 1.5 h, and washed three times with sterile water to obtain wet microspheres; the wet microspheres were then dried at 40℃ to obtain composite microbial agent slow-release microspheres. The bacterial cell concentration of the composite microbial agent suspension is 5 × 10⁻⁶. 8 CFU / mL; the compound microbial agent includes purple non-sulfur bacteria, Pseudomonas aeruginosa, and yeast; the volume ratio of the purple non-sulfur bacteria solution, Pseudomonas aeruginosa solution, and yeast solution is 2.5:1:1.
[0020] Preparation Example 1.2 S1. Add sodium alginate to deionized water and stir at 60°C until dissolved to obtain a 3% sodium alginate solution; Add potato starch to deionized water, stir well, then add boiling water and heat and stir until transparent to obtain a starch solution with a concentration of 1.5%. Sodium carboxymethyl cellulose was added to deionized water, allowed to stand for 2 hours, and then stirred until homogeneous to obtain a sodium carboxymethyl cellulose solution with a concentration of 0.75%. After grinding the meat and bone meal into powder, it was passed through a 100-mesh sieve, added to deionized water, and ultrasonically treated at 60°C for 30 minutes. Then, it was filtered to remove insoluble matter, resulting in a meat and bone meal solution with a concentration of 7.5%. Sodium alginate solution, starch solution, sodium carboxymethyl cellulose solution, and meat and bone meal solution were mixed in a volume ratio of 1.9:1.1:1.1:1 and stirred at 65°C for 1.5 hours. The pH was then adjusted to 6.8 with 1M sodium hydroxide solution to obtain the composite wall material solution. S2. The composite microbial agent suspension and the composite wall material solution were mixed evenly at a volume ratio of 1:2.25; the mixture was then shaken and cultured in a shaker at 34℃ and 190 rpm for 1.5 h; the resulting mixed solution was then injected into 200 mL of 2% calcium chloride solution for cross-linking for 2 h, and washed three times with sterile water to obtain wet microspheres; the wet microspheres were then dried at 43℃ to obtain composite microbial agent sustained-release microspheres. The bacterial cell concentration of the composite microbial agent suspension is 7.5 × 10⁻⁶. 8CFU / mL; the compound microbial agent includes purple non-sulfur bacteria, Pseudomonas aeruginosa, and yeast; the volume ratio of the purple non-sulfur bacteria solution, Pseudomonas aeruginosa solution, and yeast solution is 2.75:1.25:1.
[0021] Preparation Example 1.3 S1. Add sodium alginate to deionized water and stir at 70°C until dissolved to obtain a 4% sodium alginate solution; Add potato starch to deionized water, stir well, then add boiling water and heat and stir until transparent to obtain a starch solution with a concentration of 2%. Sodium carboxymethyl cellulose was added to deionized water, allowed to stand for 2 hours, and then stirred until homogeneous to obtain a 1% sodium carboxymethyl cellulose solution. After grinding the meat and bone meal into powder, it was passed through a 100-mesh sieve, added to deionized water, and ultrasonically treated at 70°C for 40 minutes. The insoluble matter was then removed by filtration to obtain a 10% meat and bone meal solution. Sodium alginate solution, starch solution, sodium carboxymethyl cellulose solution, and meat and bone meal solution were mixed in a volume ratio of 2:1.2:1.2:1 and stirred at 70°C for 2 hours. The pH was then adjusted to 7 with 1M sodium hydroxide solution to obtain the composite wall material solution. S2. The composite microbial agent suspension and the composite wall material solution were mixed evenly at a volume ratio of 1:2.5; the mixture was then shaken and cultured on a shaker at 36℃ and 200rpm for 1h; the resulting mixed solution was then injected into 200mL of 2% calcium chloride solution for cross-linking for 2.5h, and washed 3 times with sterile water to obtain wet microspheres; the wet microspheres were then dried at 40℃ to obtain composite microbial agent slow-release microspheres. The bacterial cell concentration of the composite microbial agent suspension is 10 × 10⁻⁶. 8 CFU / mL; the compound microbial agent includes purple non-sulfur bacteria, Pseudomonas aeruginosa, and yeast; the volume ratio of the purple non-sulfur bacteria solution, Pseudomonas aeruginosa solution, and yeast solution is 3:1.5:1.
[0022] Preparation Example 2: Jasmonic Acid Sustained-Release Microspheres Preparation Example 2.1 S1. Add 20g of potato starch to 200g of citrate-sodium dihydrogen phosphate buffer solution and stir to obtain a starch slurry solution; preheat at 50℃ for 20min, then add 2.8g of complex enzyme and react at 50℃ for 7h; after the reaction, centrifuge the obtained product, wash with distilled water, freeze-dry for 12h, and pass through a 100-mesh sieve to obtain porous starch; the complex enzyme includes α-amylase and saccharifying enzyme in a mass ratio of 1:1.5; S2. Add 10g of porous starch prepared by S1 to 100g of ethanol solution, stir evenly to obtain starch slurry solution, preheat at 30℃ for 20min, add 12g of sodium hydroxide, alkalize at 30℃ for 30min, raise the temperature to 45℃, add 58g of sodium chloroacetate and 4g of sodium hydroxide, etherify at 45℃ for 3h; then adjust the pH of starch slurry solution to 6.5 to end the etherification reaction; after centrifugation and washing with 85% ethanol solution, dry the product at 40℃ overnight, pass through a 100-mesh sieve to obtain carboxymethyl porous starch; S3. Add 50g of carboxymethyl porous starch and 5g of sodium alginate to water and stir thoroughly until dissolved to obtain a composite wall material solution; add 10g of jasmonic acid to 40g of ethanol and stir magnetically until dissolved to obtain a jasmonic acid solution; mix the composite wall material solution and the jasmonic acid solution and sonicate at 60℃ for 30min to obtain a mixed solution; stir 6g of Span-80 in 240g of corn oil until homogeneous, then add to the mixed solution, shear emulsify at 8000rpm for 10min, add 200g of 2% calcium chloride solution for crosslinking reaction for 30min, centrifuge and wash to obtain wet microspheres; freeze-dry to obtain jasmonic acid sustained-release microspheres.
[0023] Preparation Example 2.2 S1. Add 25g of potato starch to 250g of citrate-sodium dihydrogen phosphate buffer solution and stir to obtain a starch slurry solution; preheat at 55℃ for 25min, then add 3.5g of compound enzyme and react at 55℃ for 8h; after the reaction, centrifuge the obtained product, wash with distilled water, freeze-dry for 18h, and pass through a 100-mesh sieve to obtain porous starch; the compound enzyme includes α-amylase and saccharifying enzyme in a mass ratio of 1:1.6; S2. Add 12.5g of porous starch prepared by S1 to 125g of ethanol solution, stir evenly to obtain starch slurry solution, preheat at 35℃ for 23min, add 15g of sodium hydroxide, alkalize at 35℃ for 35min, raise the temperature to 50℃, add 72.5g of sodium chloroacetate and 5g of sodium hydroxide, etherify at 50℃ for 4h; then adjust the pH of starch slurry solution to 6.8 to end the etherification reaction; after centrifugation and washing with 85% ethanol solution, dry the product at 45℃ overnight, pass through a 100-mesh sieve to obtain carboxymethyl porous starch; S3. Add 62.5g of carboxymethyl porous starch and 6.25g of sodium alginate to water and stir thoroughly until dissolved to obtain a composite wall material solution; add 12.5g of jasmonic acid to 50g of ethanol and stir magnetically until dissolved to obtain a jasmonic acid solution; mix the composite wall material solution and the jasmonic acid solution and sonicate at 65℃ for 40min to obtain a mixed solution; stir 7.5g of Span-80 in 300g of corn oil until homogeneous, then add to the mixed solution, shear emulsify at 9000rpm for 15min, then add 250g of 2% calcium chloride solution for crosslinking reaction for 40min, centrifuge and wash to obtain wet microspheres; freeze-dry to obtain jasmonic acid sustained-release microspheres.
[0024] Preparation Example 2.3 S1. Add 30g of potato starch to 300g of citrate-sodium dihydrogen phosphate buffer solution and stir to obtain a starch slurry solution; preheat at 60℃ for 30min, then add 4.2g of complex enzyme and react at 60℃ for 9h; after the reaction, centrifuge the obtained product, wash with distilled water, freeze-dry for 24h, and pass through a 100-mesh sieve to obtain porous starch; the complex enzyme includes α-amylase and saccharifying enzyme in a mass ratio of 1:1.7; S2. Add 15g of porous starch prepared by S1 to 150g of ethanol solution, stir evenly to obtain starch slurry solution, preheat at 40℃ for 25min, add 18g of sodium hydroxide, alkalize at 40℃ for 40min, raise the temperature to 55℃, add 87g of sodium chloroacetate and 6g of sodium hydroxide, etherify at 55℃ for 5h; then adjust the pH of starch slurry solution to 7 to end the etherification reaction; after centrifugation and washing with 85% ethanol solution, dry the product at 50℃ overnight, pass through a 100-mesh sieve to obtain carboxymethyl porous starch; S3. Add 75g of carboxymethyl porous starch and 7.5g of sodium alginate to water and stir thoroughly until dissolved to obtain a composite wall material solution; add 15g of jasmonic acid to 60g of ethanol and stir magnetically until dissolved to obtain a jasmonic acid solution; mix the composite wall material solution and the jasmonic acid solution and sonicate at 70℃ for 50min to obtain a mixed solution; stir 9g of Span-80 in 360g of corn oil until homogeneous, then add to the mixed solution, shear emulsify at 10000rpm for 20min, then add 300g of 2% calcium chloride solution for crosslinking reaction for 50min, centrifuge and wash to obtain wet microspheres; freeze-dry to obtain jasmonic acid sustained-release microspheres.
[0025] Example 1 Add 0.5g sodium nitroprusside, 2g potassium dihydrogen phosphate, 0.4g hydrogen peroxide, and 10g humic acid to 1000g water in sequence, and stir until completely dissolved to obtain mixed solution a; Add 0.2g gibberellin, 0.2g 6-benzylaminopurine, and 0.2g indolebutyric acid to 60g ethanol and stir until completely dissolved to obtain mixed solution b; After mixing solutions a and b thoroughly, while maintaining stirring, add 2g of seaweed extract, 5g of compound amino acids, 1g of jasmonic acid sustained-release microspheres prepared in Preparation Example 2.1, and 1g of compound microbial agent sustained-release microspheres prepared in Preparation Example 1.1 in sequence. Continue stirring for 30 minutes to obtain the underground propagation complex activator for ecological restoration. The compound amino acids include proline, tryptophan, and valine in a mass ratio of 2:1:1.
[0026] Example 2 Add 0.6g sodium nitroprusside, 2.5g potassium dihydrogen phosphate, 0.45g hydrogen peroxide, and 11.5g humic acid to 1000g of water in sequence, and stir until completely dissolved to obtain mixed solution a; Add 0.25g gibberellin, 0.35g 6-benzylaminopurine, and 0.25g indolebutyric acid to 67g ethanol and stir until completely dissolved to obtain mixed solution b; After mixing solutions a and b thoroughly, while maintaining stirring, add 2.5g of seaweed extract, 6g of compound amino acids, 1g of jasmonic acid sustained-release microspheres prepared in Preparation Example 2.1, and 1g of compound microbial agent sustained-release microspheres prepared in Preparation Example 1.1 in sequence. Continue stirring for 45 minutes to obtain the underground propagation compound activator for ecological restoration. The compound amino acids include proline, tryptophan, and valine in a mass ratio of 2:1:1.
[0027] Example 3 Add 0.7g sodium nitroprusside, 3g potassium dihydrogen phosphate, 0.5g hydrogen peroxide, and 13g humic acid to 1000g of water in sequence, and stir until completely dissolved to obtain mixed solution a; Add 0.3g gibberellin, 0.5g 6-benzylaminopurine, and 0.3g indolebutyric acid to 74g ethanol and stir until completely dissolved to obtain mixed solution b; After mixing solutions a and b thoroughly, while maintaining stirring, add 3g of seaweed extract, 7g of compound amino acids, 1g of jasmonic acid sustained-release microspheres prepared in Preparation Example 2.1, and 1g of compound microbial agent sustained-release microspheres prepared in Preparation Example 1.1 in sequence. Continue stirring for 60 minutes to obtain the underground propagation complex activator for ecological restoration. The compound amino acids include proline, tryptophan, and valine in a mass ratio of 2:1:1.
[0028] Example 4 The difference between Example 4 and Example 1 is that the mass ratio of proline, tryptophan and valine in the composite amino acid used in Example 4 is 2.5:1.5:1.
[0029] Example 5 The difference between Example 5 and Example 1 is that the mass ratio of proline, tryptophan, and valine in the composite amino acid used in Example 5 is 3:2:1.
[0030] Example 6 The difference between Example 6 and Example 1 is that the mass ratio of proline, tryptophan and valine in the composite amino acid used in Example 6 is 1:0.5:1.
[0031] Example 7 The difference between Example 7 and Example 1 is that the mass ratio of proline, tryptophan and valine in the composite amino acid used in Example 7 is 4:2.5:1.
[0032] Example 8 The difference between Example 8 and Example 1 is that the mass of the jasmonic acid sustained-release microspheres prepared in Example 2.1 used in Example 8 is 1.5g.
[0033] Example 9 The difference between Example 9 and Example 1 is that the mass of the jasmonic acid sustained-release microspheres prepared in Example 2.1 used in Example 9 is 2g.
[0034] Example 10 The difference between Example 10 and Example 1 is that the jasmonic acid sustained-release microspheres used in Example 10 were prepared from Preparation Example 2.2.
[0035] Example 11 The difference between Example 11 and Example 1 is that the jasmonic acid sustained-release microspheres used in Example 11 were prepared from Preparation Example 2.3.
[0036] Example 12 The difference between Example 12 and Example 1 is that the mass of the composite microbial agent sustained-release microspheres prepared in Example 1.1 used in Example 12 is 2g.
[0037] Example 13 The difference between Example 13 and Example 1 is that the mass of the composite microbial agent sustained-release microspheres prepared in Example 1.1 used in Example 13 is 3g.
[0038] Example 14 The difference between Example 14 and Example 1 is that the composite microbial agent slow-release microspheres used in Example 14 were prepared from Preparation Example 1.2.
[0039] Example 15 The difference between Example 15 and Example 1 is that the composite microbial agent slow-release microspheres used in Example 15 were prepared from Preparation Example 1.3.
[0040] Comparative Example 1 The difference between Comparative Example 1 and Example 1 is that jasmonic acid sustained-release microspheres were not added in Comparative Example 1.
[0041] Comparative Example 2 The difference between Comparative Example 2 and Example 1 is that the mass of the jasmonic acid sustained-release microspheres prepared in Comparative Example 2 is 3g.
[0042] Comparative Example 3 The difference between Comparative Example 3 and Example 1 is that in Comparative Example 3, an equal amount of jasmonic acid was used instead of the jasmonic acid sustained-release microspheres prepared in Preparation Example 2.1.
[0043] Comparative Example 4 The difference between Comparative Example 4 and Example 1 is that no composite microbial agent slow-release microspheres were added in Comparative Example 4.
[0044] Comparative Example 5 The difference between Comparative Example 5 and Example 1 is that the mass of the composite microbial agent sustained-release microspheres prepared in Comparative Example 5 from Preparation Example 1.1 is 5g.
[0045] Performance testing Experiment location: grassland of the hay storage station in Duolun County.
[0046] Grassland type: hayfield.
[0047] Application period: Spring soil thawing period, soil thawing depth 5cm.
[0048] Using a no-till planter, the underground propagule compound activator of Examples 1-15 and Comparative Examples 1-5 was applied in strips to a soil depth of 3-4 cm and a row spacing of 15 cm. Each experimental grassland area was 5 mu (approximately 0.33 hectares), and three replicates were set up. A control group without any grassland conditioner was also included.
[0049] Application rate: 15 kg / mu. Post-application management: After treatment, the natural grassland should be mowed in the fall according to traditional methods.
[0050] Results: The forage yield and root biomass of each group were measured over 5 years, and the results are shown in Table 1.
[0051] The specific test results are as follows:
[0052] As can be seen from the test results in Table 1, the underground propagule composite activator for ecological restoration and its preparation method provided in this application greatly improves grassland forage yield and increases underground root biomass.
[0053] This specific embodiment is merely an explanation of this application and is not intended to limit it. After reading this specification, those skilled in the art can make modifications to this embodiment without contributing any inventive step, but such modifications are protected by patent law as long as they fall within the scope of the claims of this application.
Claims
1. An underground propagule composite inoculant for ecological restoration, characterized by: The raw materials include 0.2-0.3 parts of gibberellin, 0.2-0.5 parts of 6-benzylaminopurine, 0.2-0.3 parts of indolebutyric acid, 0.5-0.7 parts of sodium nitroprusside, 0.4-0.5 parts of hydrogen peroxide, 1-2 parts of jasmonic acid slow-release microspheres, 5-7 parts of compound amino acid, 1-3 parts of compound microbial agent slow-release microspheres, 2-3 parts of seaweed extract, 10-13 parts of humic acid, 2-3 parts of potassium dihydrogen phosphate, 60-74 parts of ethanol, and 1000 parts of water.
2. The underground propagule composite inoculant for ecological restoration according to claim 1, characterized in that: The compound amino acid includes proline, tryptophan and valine, and the mass ratio of the proline, tryptophan and valine is (2-3):(1-2):
1.
3. The underground propagule composite inoculant for ecological restoration according to claim 1, characterized in that: The preparation method of the compound microbial agent slow-release microspheres comprises the following steps: The compound microbial agent suspension and the compound wall material solution are mixed uniformly at a volume ratio of 1:(2-2.5), and then are subjected to shaking culture at 32-36 DEG C and 180-200 rpm for 1-2 h; then the obtained mixed solution is injected into a 2% calcium chloride solution for crosslinking for 1.5-2.5 h, and then is washed with sterile water for several times to obtain wet microspheres; the wet microspheres are dried at 40-45 DEG C to obtain the compound microbial agent slow-release microspheres.
4. The underground propagule composite inoculant for ecological restoration according to claim 3, characterized in that: The bacterial concentration of the complex microbial agent suspension is 5×10 8 -10×10 8 CFU / mL; the complex microbial agent comprises purple non-sulfur bacteria, pseudomonas aeruginosa and yeast; the volume ratio of the purple non-sulfur bacteria solution, the pseudomonas aeruginosa solution and the yeast solution is (2.5-3):(1-1.5):
1.
5. The underground propagule composite inoculant for ecological restoration according to claim 3, characterized in that: The compound wall material includes sodium alginate, starch, sodium carboxymethyl cellulose and meat and bone meal; The preparation method of the compound wall material solution comprises the following steps: Sodium alginate is added into water, and is stirred to dissolve at 60-70 DEG C to obtain a 2-4% sodium alginate solution; Starch is added into water, and is stirred to dissolve, and then boiling water is added, and the mixture is heated and stirred to be transparent to obtain a 1-2% starch solution; Sodium carboxymethyl cellulose is added into water, and is stirred to dissolve after being placed for 2-3 h to obtain a 0.5-1% sodium carboxymethyl cellulose solution; Meat and bone meal is crushed, is sieved, is added into water, and is ultrasonically treated at 60-70 DEG C for 30-40 min, and then is filtered to remove insoluble substances to obtain a 5-10% meat and bone meal solution; The sodium alginate solution, the starch solution, the sodium carboxymethyl cellulose solution and the meat and bone meal solution are mixed at a volume ratio of (1.8-2):(1-1.2):(1-1.2):1, and then are stirred at 60-70 DEG C for 1-2 h, and then the pH value is adjusted to 6.5-7.0 to obtain the compound wall material solution.
6. The underground propagule composite inoculant for ecological restoration according to claim 1, characterized in that: The jasmonic acid slow-release microspheres are prepared from the following raw materials by weight: 10-15 parts of jasmonic acid, 50-75 parts of carboxymethyl porous starch, 5-7.5 parts of sodium alginate, 200-300 parts of water, 40-60 parts of ethanol, 200-300 parts of 2% calcium chloride solution, 240-360 parts of corn oil and 6-9 parts of emulsifier.
7. The underground propagule composite inoculant for ecological restoration according to claim 6, characterized in that: The carboxymethyl porous starch is prepared from the following raw materials by weight: 10-15 parts of porous starch, 100-150 parts of ethanol solution, 16-24 parts of sodium hydroxide and 58-87 parts of sodium chloroacetate; The porous starch is prepared from the following raw materials by weight: 20-30 parts of starch, 200-300 parts of citric acid-sodium dihydrogen phosphate buffer and 2.8-4.2 parts of compound enzyme. The complex enzyme comprises alpha-amylase and saccharifying enzyme, and the mass ratio is 1: (1.5-1.7).
8. The underground propagule composite inoculant for ecological restoration according to claim 7, characterized in that: The preparation method of the carboxymethyl porous starch comprises the following steps: According to weight parts, 10-15 parts of porous starch are added into 100-150 parts of ethanol solution, and stirred uniformly to obtain a starch milk solution; after preheating at 30-40 ℃ for 20-25 min, 12-18 parts of sodium hydroxide is added, and alkali reaction is carried out at 30-40 ℃ for 30-40 min; then, the temperature is increased to 45-55 ℃, 58-87 parts of sodium chloroacetate and 4-6 parts of sodium hydroxide are added, and etherification is carried out at 45-55 ℃ for 3-5 h; then, the pH of the starch milk solution is adjusted to 6.5-7.0 to end the etherification reaction; after centrifugation and washing, the product is dried at 40-50 ℃ overnight, and sieved to obtain the carboxymethyl porous starch. The preparation method of the porous starch comprises the following steps: According to weight parts, 20-30 parts of starch are added into 200-300 parts of a citric acid-sodium dihydrogen phosphate buffer solution, and stirred to obtain a starch milk solution; after preheating at 50-60 ℃ for 20-30 min, 2.8-4.2 parts of a complex enzyme is added, and reaction is carried out at 50-60 ℃ for 7-9 h; after the reaction is completed, the obtained product is subjected to centrifugation, distilled water washing, and freeze-drying for 12-24 h, and then sieved to obtain the porous starch.
9. The underground propagule composite inoculant for ecological restoration according to claim 6, characterized in that: The preparation method of the jasmonic acid sustained-release microspheres comprises the following steps: Carboxymethyl porous starch and sodium alginate are added into water, and fully stirred until dissolved to obtain a composite wall material solution; jasmonic acid is added into ethanol, and magnetically stirred until dissolved to obtain a jasmonic acid solution; the composite wall material solution and the jasmonic acid solution are mixed, and then ultrasonically treated at 60-70 ℃ for 30-50 min to obtain a mixed solution; an emulsifier is uniformly stirred in corn oil, and then added into the mixed solution; after shearing emulsification at 8000-10000 rpm for 10-20 min, a calcium chloride solution with a concentration of 2% is added for crosslinking reaction for 30-50 min; after centrifugation and washing, wet microspheres are obtained; after freeze-drying, the jasmonic acid sustained-release microspheres are obtained.
10. The method for preparing the underground propagule composite inoculant for ecological restoration according to any one of claims 1-9, characterized in that: The preparation method comprises the following steps: Sodium nitroprusside, potassium dihydrogen phosphate, hydrogen peroxide and humic acid are sequentially added into water, and stirred until completely dissolved to obtain a mixed solution a; Gibberellin, 6-benzylaminopurine and indolebutyric acid are added into ethanol, and stirred until completely dissolved to obtain a mixed solution b; After the mixed solution a and the mixed solution b are uniformly mixed, stirring is kept, and seaweed extract, complex amino acid, jasmonic acid sustained-release microspheres and complex microbial agent sustained-release microspheres are sequentially added; after continuous stirring for 30-60 min, the underground propagulum composite elicitor for ecological restoration is obtained.