CN111848240A - Microbial agent capable of promoting flower bud differentiation of plants - Google Patents
Microbial agent capable of promoting flower bud differentiation of plants Download PDFInfo
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- CN111848240A CN111848240A CN202010766225.3A CN202010766225A CN111848240A CN 111848240 A CN111848240 A CN 111848240A CN 202010766225 A CN202010766225 A CN 202010766225A CN 111848240 A CN111848240 A CN 111848240A
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Abstract
The invention discloses a microbial agent capable of promoting flower bud differentiation of plants, and belongs to the technical field of microbial agents. The microbial inoculum comprises a component A and a component B, wherein the component A is prepared from the following raw materials in parts by weight: 10-20 parts of bacillus subtilis, 5-10 parts of bacillus licheniformis, 5-10 parts of lactobacillus plantarum, 5-8 parts of bacillus megaterium, 3-5 parts of bacillus mucilaginosus and 1-3 parts of a dispersing agent; the component B is 5-10 parts of gibberellin-indoleacetic acid-protein complex. The addition of the dispersing agent and the protein compound provided by the invention have synergistic effect, so that effective flower bud differentiation of crops is realized together, the great increase in yield and income of vegetables and fruits such as tomatoes can be realized, the quality of the crops is improved, and the economic benefit is remarkable.
Description
Technical Field
The invention belongs to the technical field of microbial preparations, and particularly relates to a microbial preparation capable of promoting flower bud differentiation of plants.
Background
Flower bud differentiation is a key stage in the transition from vegetative to reproductive growth of plants and is a complex morphogenetic process. The flower bud differentiation of the plant is not only influenced by environmental factors, but also is under the combined action and regulation of various internal factors (saccharides, endogenous hormones and the like), wherein the endogenous hormones have close relation with the flower bud differentiation of the plant. The differentiation of the plant flower buds is divided into a physiological differentiation period and a morphological differentiation period, the physiological differentiation is the basis of morphological differentiation, the morphological differentiation of the flower buds is influenced by the content level of endogenous hormones in the physiological differentiation period, and the morphological establishment of the flower buds is influenced by the content level of the endogenous hormones in the morphological differentiation period.
At present, methods for promoting plant flower bud differentiation are not limited to methods for adjusting planting methods, spraying plant hormones and the like, the defects of slow effect taking and instability existing in adjusting planting are overcome, the fruit quality can be affected to a certain extent by spraying a large amount of plant hormones, the plants can generate certain stress resistance after long-term use, the stability of the soil environment is not facilitated, and the method is not beneficial to sustainable development.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides the microbial agent which can greatly promote the flower bud differentiation of plants, greatly reduce the addition of plant hormones through the high-efficiency slow release effect, and simultaneously can adjust the soil environment and protect the soil ecological environment.
The technical means adopted by the invention are as follows:
a microbial agent capable of promoting flower bud differentiation of plants comprises a component A and a component B, wherein the component A is prepared from the following raw materials in parts by weight: 10-20 parts of bacillus subtilis, 5-10 parts of bacillus licheniformis, 5-10 parts of lactobacillus plantarum, 5-8 parts of bacillus megaterium, 3-5 parts of bacillus mucilaginosus and 1-3 parts of a dispersing agent; the component B is 5-10 parts of gibberellin-indoleacetic acid-protein complex and is prepared by the following steps:
1) drying peanut kernels, removing red skins, crushing, sieving with a 60-mesh sieve, adding petroleum ether with the mass volume of 5 times, degreasing and leaching for 2 hours, centrifuging for 5 minutes at 4 ℃ at 6000r/min, repeatedly leaching and degreasing for three times, centrifuging to obtain solid residues, adding 10% NaOH according to the mass ratio of 1: 5, performing size mixing, adjusting the pH to 9, stirring, centrifuging for 5 minutes at 6000r/min, collecting filtrate, adjusting the pH of the filtrate to 4 with 8% HCl, standing for 1 hour, collecting precipitates, washing the precipitates with deionized water to be neutral, freezing and storing, and drying for 72 hours with a vacuum freeze dryer to obtain proteins for later use;
2) weighing 0.5g of gibberellin and 0.1g of indoleacetic acid, dissolving in 1L of absolute ethanol to prepare a gibberellin-indoleacetic acid ethanol solution, taking 30mL of gibberellin-indoleacetic acid ethanol solution and 5g of protein prepared in the step (1), uniformly mixing, stirring in the dark for reaction for 2h, and then adding 5mmol/L of CaCl into the mixed solution280mL, standing at room temperature for 16h, desalting with 14000D dialysis bag for 72h, and freeze-drying to obtain gibberellin-indoleacetic acid protein complex.
The preferable microbial agent capable of promoting flower bud differentiation of plants comprises a component A and a component B, wherein the component A is prepared from the following raw materials in parts by weight: 20 parts of bacillus subtilis, 10 parts of bacillus licheniformis, 10 parts of lactobacillus plantarum, 8 parts of bacillus megaterium, 5 parts of bacillus mucilaginosus and 3 parts of a dispersing agent; the component B is 10 parts of gibberellin-protein compound
Preferably, the dispersing agent is obtained by mixing sodium lignosulphonate and chitosan according to the mass ratio of 1: 1.
The invention relates to a microbial agent for promoting plant flower bud differentiation, which is prepared by the following steps:
1) inoculating bacillus subtilis into a liquid culture medium, and culturing for 25h at the condition of 30 ℃ and at the rotating speed of a shaker of 200r/min to obtain a bacillus subtilis seed solution; inoculating the bacillus subtilis seed solution into a 500L seed tank culture medium with the inoculation amount of 8%, standing and culturing for 30h at the temperature of 30 ℃, and centrifugally collecting thalli; spray-freezing and drying the thalli to obtain bacillus subtilis;
2) respectively preparing bacillus licheniformis, lactobacillus plantarum, bacillus megaterium and bacillus mucilaginosus according to the method in the step (1);
3) uniformly mixing bacillus subtilis, bacillus licheniformis, lactobacillus plantarum, bacillus megaterium, bacillus mucilaginosus and a dispersing agent according to a weight ratio, adding the mixture into an airflow crusher, controlling the fineness to be 800 meshes, and drying to obtain a component A;
4) b, preparation of a component: drying peanut kernels, removing red skins, crushing, sieving with a 60-mesh sieve, adding petroleum ether with the mass volume of 5 times, degreasing and leaching for 2 hours, centrifuging for 5 minutes at 4 ℃ at 6000r/min, repeatedly leaching and degreasing for three times, centrifuging to obtain solid residues, adding 10% NaOH according to the mass ratio of 1: 5, performing size mixing, adjusting the pH to 9, stirring, centrifuging for 5 minutes at 6000r/min, collecting filtrate, adjusting the pH of the filtrate to 4 with 8% HCl, standing for 1 hour, collecting precipitates, washing the precipitates with deionized water to be neutral, freezing and storing, and drying for 72 hours with a vacuum freeze dryer to obtain proteins for later use; weighing 0.5g of gibberellin and 0.1g of indoleacetic acid, dissolving in 1L of absolute ethanol to prepare a gibberellin-indoleacetic acid ethanol solution, taking 30mL of gibberellin-indoleacetic acid ethanol solution and 5g of protein, uniformly mixing, stirring in the dark for reaction for 2h, and then adding 5mmol/L of CaCl into the mixed solution280mL, standing at room temperature for 16h, desalting with 14000D dialysis bag for 72h, and freeze-drying to obtain gibberellin-indoleacetic acid protein complex.
The liquid culture medium comprises the following components: weighing 25g of bean sprouts, adding 75mL of water, boiling for 20min, filtering and removing residues, wherein the volume is 100 mL; adding 5g glucose, dissolving, mixing, adjusting pH to 7.0, and sterilizing at 121 deg.C for 30 min.
The specific composition of the culture medium in the seeding tank is as follows: 0.3% of beef extract, 1% of peptone, 0.5% of sodium chloride and 0.5% of glucose.
The bacillus subtilis, the bacillus licheniformis, the lactobacillus plantarum, the bacillus megaterium and the bacillus mucilaginosus are all sold in the market, and the effective viable count is 100-200 billion/g.
And (4) during packaging, storing the component B in a vacuum and dark state.
When in use, the component B is dissolved in 10 times of aqueous solution, and the component A is added and mixed evenly for use. When the biological agent is actually used, the biological agent is diluted by 500-800 times and is sprayed for use.
Advantageous effects
The bacillus subtilis is used, can generate various organic acids and inorganic acids, reduces the pH value in the environment, degrades insoluble phosphate into effective phosphorus, and promotes crops to absorb P. Meanwhile, the bacillus mucilaginosus can decompose silicate minerals in the soil, so that insoluble potassium, phosphorus, silicon and the like in the soil are converted into soluble substances for plant growth and utilization. The bacillus licheniformis and the bacillus megatherium can generate resistance substances such as lipopeptide, peptide, phospholipid, bacteriocin and the like, improve the disease resistance of crops, and provide a foundation for the differentiation of flower buds of later-stage crops. The invention selects the mutual coordination and proliferation relation of microbial strains to form a complex biological flora with various bacterial microbial communities, stable structure and wide functions, and the biological flora is quickly combined with the benign force in soil to generate antioxidant substances, remove the oxidant substances, inhibit pathogenic bacteria, form a good environment suitable for the growth of crops, improve the disease resistance of the crops, and simultaneously generate a large amount of beneficial substances easily absorbed by the crops to promote the healthy growth of the crops and improve the quality of the crops.
According to the invention, the compound microbial agent ensures the healthy growth state of crops, the plant hormone is used for regulating the flower bud differentiation of plants, a large amount of plant hormone is not only directly sprayed, but is wrapped in the plant protein, so that the plant hormone can be continuously and slowly released, the differentiation and growth of the plants are effectively promoted, and the use amount of the plant hormone is reduced to a certain extent while the plant hormone is protected from loss. The protein complex is quickly activated after being dissolved in water, is mixed with the component A, and then is quickly adsorbed on the surface of protein under the action of the dispersing agent, so that the microbial agent has the effects of promoting growth and resisting diseases after being applied to crops, and the protein complex simultaneously releases phytohormone. The addition of the dispersing agent and the synergistic effect of the protein compound can realize effective flower bud differentiation of crops, can realize great increase in yield and income of vegetables and fruits such as tomatoes and the like, and can improve the quality of the crops, and the economic benefit is remarkable.
Detailed Description
The technical solution of the present invention is further described below with reference to specific embodiments, but is not limited thereto.
Example 1
A microbial agent capable of promoting flower bud differentiation of plants comprises a component A and a component B, wherein the component A is prepared from the following raw materials in parts by weight: 10 parts of bacillus subtilis, 5 parts of bacillus licheniformis, 5 parts of lactobacillus plantarum, 5 parts of bacillus megaterium, 3 parts of bacillus mucilaginosus and 1 part of dispersing agent; the component B is 5 parts of gibberellin-indoleacetic acid-protein complex and is prepared by the following steps:
1) drying peanut kernels, removing red skins, crushing, sieving with a 60-mesh sieve, adding petroleum ether with the mass volume of 5 times, degreasing and leaching for 2 hours, centrifuging for 5 minutes at 4 ℃ at 6000r/min, repeatedly leaching and degreasing for three times, centrifuging to obtain solid residues, adding 10% NaOH according to the mass ratio of 1: 5, performing size mixing, adjusting the pH to 9, stirring, centrifuging for 5 minutes at 6000r/min, collecting filtrate, adjusting the pH of the filtrate to 4 with 8% HCl, standing for 1 hour, collecting precipitates, washing the precipitates with deionized water to be neutral, freezing and storing, and drying for 72 hours with a vacuum freeze dryer to obtain proteins for later use;
2) weighing 0.5g of gibberellin and 0.1g of indoleacetic acid, dissolving in 1L of absolute ethanol to prepare a gibberellin-indoleacetic acid ethanol solution, taking 30mL of gibberellin-indoleacetic acid ethanol solution and 5g of protein prepared in the step (1), uniformly mixing, stirring in the dark for reaction for 2h, and then adding 5mmol/L of CaCl into the mixed solution280mL, standing at room temperature for 16h, desalting with 14000D dialysis bag for 72h, and freeze-drying to obtain gibberellin-indoleacetic acid protein complex.
The dispersing agent is obtained by mixing sodium lignosulphonate and chitosan according to the mass ratio of 1: 1.
The invention relates to a microbial agent for promoting plant flower bud differentiation, which is prepared by the following steps:
1) inoculating bacillus subtilis into a liquid culture medium, and culturing for 25h at the condition of 30 ℃ and at the rotating speed of a shaker of 200r/min to obtain a bacillus subtilis seed solution; inoculating the bacillus subtilis seed solution into a 500L seed tank culture medium with the inoculation amount of 8%, standing and culturing for 30h at the temperature of 30 ℃, and centrifugally collecting thalli; spray-freezing and drying the thalli to obtain bacillus subtilis;
2) respectively preparing bacillus licheniformis, lactobacillus plantarum, bacillus megaterium and bacillus mucilaginosus according to the method in the step (1);
3) uniformly mixing bacillus subtilis, bacillus licheniformis, lactobacillus plantarum, bacillus megaterium, bacillus mucilaginosus and a dispersing agent according to a weight ratio, adding the mixture into an airflow crusher, controlling the fineness to be 800 meshes, and drying to obtain a component A;
4) b, preparation of a component: drying peanut kernels, removing red skins, crushing, sieving with a 60-mesh sieve, adding petroleum ether with the mass volume of 5 times, degreasing and leaching for 2 hours, centrifuging for 5 minutes at 4 ℃ at 6000r/min, repeatedly leaching and degreasing for three times, centrifuging to obtain solid residues, adding 10% NaOH according to the mass ratio of 1: 5, performing size mixing, adjusting the pH to 9, stirring, centrifuging for 5 minutes at 6000r/min, collecting filtrate, adjusting the pH of the filtrate to 4 with 8% HCl, standing for 1 hour, collecting precipitates, washing the precipitates with deionized water to be neutral, freezing and storing, and drying for 72 hours with a vacuum freeze dryer to obtain proteins for later use; weighing 0.5g of gibberellin and 0.1g of indoleacetic acid, dissolving in 1L of absolute ethanol to prepare a gibberellin-indoleacetic acid ethanol solution, taking 30mL of gibberellin-indoleacetic acid ethanol solution and 5g of protein, uniformly mixing, stirring in the dark for reaction for 2h, and then adding 5mmol/L of CaCl into the mixed solution280mL, standing at room temperature for 16h, desalting with 14000D dialysis bag for 72h, and freeze-drying to obtain gibberellin-indoleacetic acid protein complex.
The liquid culture medium comprises the following components: weighing 25g of bean sprouts, adding 75mL of water, boiling for 20min, filtering and removing residues, wherein the volume is 100 mL; adding 5g glucose, dissolving, mixing, adjusting pH to 7.0, and sterilizing at 121 deg.C for 30 min.
The specific composition of the culture medium in the seeding tank is as follows: 0.3% of beef extract, 1% of peptone, 0.5% of sodium chloride and 0.5% of glucose.
The bacillus subtilis, the bacillus licheniformis, the lactobacillus plantarum, the bacillus megaterium and the bacillus mucilaginosus are all sold in the market, and the effective viable count is 100-200 billion/g.
And (4) during packaging, storing the component B in a vacuum and dark state.
When in use, the component B is dissolved in 10 times of aqueous solution, and the component A is added and mixed evenly for use. When the biological agent is actually used, the biological agent is diluted by 500-800 times and is sprayed for use.
Example 2
A microbial agent capable of promoting flower bud differentiation of plants comprises a component A and a component B, wherein the component A is prepared from the following raw materials in parts by weight: 15 parts of bacillus subtilis, 8 parts of bacillus licheniformis, 8 parts of lactobacillus plantarum, 7 parts of bacillus megaterium, 4 parts of bacillus mucilaginosus and 2 parts of a dispersing agent; the component B is 8 parts of gibberellin-indoleacetic acid-protein complex and is prepared by the following steps:
1) drying peanut kernels, removing red skins, crushing, sieving with a 60-mesh sieve, adding petroleum ether with the mass volume of 5 times, degreasing and leaching for 2 hours, centrifuging for 5 minutes at 4 ℃ at 6000r/min, repeatedly leaching and degreasing for three times, centrifuging to obtain solid residues, adding 10% NaOH according to the mass ratio of 1: 5, performing size mixing, adjusting the pH to 9, stirring, centrifuging for 5 minutes at 6000r/min, collecting filtrate, adjusting the pH of the filtrate to 4 with 8% HCl, standing for 1 hour, collecting precipitates, washing the precipitates with deionized water to be neutral, freezing and storing, and drying for 72 hours with a vacuum freeze dryer to obtain proteins for later use;
2) weighing 0.5g of gibberellin and 0.1g of indoleacetic acid, dissolving in 1L of absolute ethanol to prepare a gibberellin-indoleacetic acid ethanol solution, uniformly mixing 30mL of gibberellin-indoleacetic acid ethanol solution with 5g of protein prepared in the step (1), stirring for reaction for 2h in a dark place, adding 5mmol/L of CaCl280mL into the mixed solution, standing for 16h at room temperature, desalting for 72h by using a 14000D dialysis bag, and freeze-drying to obtain the gibberellin-indoleacetic acid protein compound.
The dispersing agent is obtained by mixing sodium lignosulphonate and chitosan according to the mass ratio of 1: 1.
The invention relates to a microbial agent for promoting plant flower bud differentiation, which is prepared by the following steps:
1) inoculating bacillus subtilis into a liquid culture medium, and culturing for 25h at the condition of 30 ℃ and at the rotating speed of a shaker of 200r/min to obtain a bacillus subtilis seed solution; inoculating the bacillus subtilis seed solution into a 500L seed tank culture medium with the inoculation amount of 8%, standing and culturing for 30h at the temperature of 30 ℃, and centrifugally collecting thalli; spray-freezing and drying the thalli to obtain bacillus subtilis;
2) respectively preparing bacillus licheniformis, lactobacillus plantarum, bacillus megaterium and bacillus mucilaginosus according to the method in the step (1);
3) uniformly mixing bacillus subtilis, bacillus licheniformis, lactobacillus plantarum, bacillus megaterium, bacillus mucilaginosus and a dispersing agent according to a weight ratio, adding the mixture into an airflow crusher, controlling the fineness to be 800 meshes, and drying to obtain a component A;
4) b, preparation of a component: drying peanut kernels, removing red skins, crushing, sieving with a 60-mesh sieve, adding petroleum ether with the mass volume of 5 times, degreasing and leaching for 2 hours, centrifuging for 5 minutes at 4 ℃ at 6000r/min, repeatedly leaching and degreasing for three times, centrifuging to obtain solid residues, adding 10% NaOH according to the mass ratio of 1: 5, performing size mixing, adjusting the pH to 9, stirring, centrifuging for 5 minutes at 6000r/min, collecting filtrate, adjusting the pH of the filtrate to 4 with 8% HCl, standing for 1 hour, collecting precipitates, washing the precipitates with deionized water to be neutral, freezing and storing, and drying for 72 hours with a vacuum freeze dryer to obtain proteins for later use; weighing 0.5g of gibberellin and 0.1g of indoleacetic acid, dissolving in 1L of absolute ethanol to prepare a gibberellin-indoleacetic acid ethanol solution, taking 30mL of gibberellin-indoleacetic acid ethanol solution and 5g of protein, uniformly mixing, stirring for 2h in a dark place, adding 5mmol/L of CaCl280mL into the mixed solution, standing for 16h at room temperature, desalting for 72h by using a 14000D dialysis bag, and freeze-drying to obtain the gibberellin-indoleacetic acid protein complex.
The liquid culture medium comprises the following components: weighing 25g of bean sprouts, adding 75mL of water, boiling for 20min, filtering and removing residues, wherein the volume is 100 mL; adding 5g glucose, dissolving, mixing, adjusting pH to 7.0, and sterilizing at 121 deg.C for 30 min.
The specific composition of the culture medium in the seeding tank is as follows: 0.3% of beef extract, 1% of peptone, 0.5% of sodium chloride and 0.5% of glucose.
The bacillus subtilis, the bacillus licheniformis, the lactobacillus plantarum, the bacillus megaterium and the bacillus mucilaginosus are all sold in the market, and the effective viable count is 100-200 billion/g.
And (4) during packaging, storing the component B in a vacuum and dark state.
When in use, the component B is dissolved in 10 times of aqueous solution, and the component A is added and mixed evenly for use. When the biological agent is actually used, the biological agent is diluted by 500-800 times and is sprayed for use.
Example 3
A microbial agent capable of promoting flower bud differentiation of plants comprises a component A and a component B, wherein the component A is prepared from the following raw materials in parts by weight: 20 parts of bacillus subtilis, 10 parts of bacillus licheniformis, 10 parts of lactobacillus plantarum, 8 parts of bacillus megaterium, 5 parts of bacillus mucilaginosus and 3 parts of a dispersing agent; the component B is 5-10 parts of gibberellin-indoleacetic acid-protein complex and is prepared by the following steps:
1) drying peanut kernels, removing red skins, crushing, sieving with a 60-mesh sieve, adding petroleum ether with the mass volume of 5 times, degreasing and leaching for 2 hours, centrifuging for 5 minutes at 4 ℃ at 6000r/min, repeatedly leaching and degreasing for three times, centrifuging to obtain solid residues, adding 10% NaOH according to the mass ratio of 1: 5, performing size mixing, adjusting the pH to 9, stirring, centrifuging for 5 minutes at 6000r/min, collecting filtrate, adjusting the pH of the filtrate to 4 with 8% HCl, standing for 1 hour, collecting precipitates, washing the precipitates with deionized water to be neutral, freezing and storing, and drying for 72 hours with a vacuum freeze dryer to obtain proteins for later use;
2) weighing 0.5g of gibberellin and 0.1g of indoleacetic acid, dissolving in 1L of absolute ethanol to prepare a gibberellin-indoleacetic acid ethanol solution, taking 30mL of gibberellin-indoleacetic acid ethanol solution and 5g of protein prepared in the step (1), uniformly mixing, stirring in the dark for reaction for 2h, and then adding 5mmol/L of CaCl into the mixed solution280mL, standing at room temperature for 16h, desalting with 14000D dialysis bag for 72h, and freeze-drying to obtain gibberellin-indoleacetic acid protein complex.
The dispersing agent is obtained by mixing sodium lignosulphonate and chitosan according to the mass ratio of 1: 1.
The invention relates to a microbial agent for promoting plant flower bud differentiation, which is prepared by the following steps:
1) inoculating bacillus subtilis into a liquid culture medium, and culturing for 25h at the condition of 30 ℃ and at the rotating speed of a shaker of 200r/min to obtain a bacillus subtilis seed solution; inoculating the bacillus subtilis seed solution into a 500L seed tank culture medium with the inoculation amount of 8%, standing and culturing for 30h at the temperature of 30 ℃, and centrifugally collecting thalli; spray-freezing and drying the thalli to obtain bacillus subtilis;
2) respectively preparing bacillus licheniformis, lactobacillus plantarum, bacillus megaterium and bacillus mucilaginosus according to the method in the step (1);
3) uniformly mixing bacillus subtilis, bacillus licheniformis, lactobacillus plantarum, bacillus megaterium, bacillus mucilaginosus and a dispersing agent according to a weight ratio, adding the mixture into an airflow crusher, controlling the fineness to be 800 meshes, and drying to obtain a component A;
4) b, preparation of a component: drying peanut kernels, removing red skins, crushing, sieving with a 60-mesh sieve, adding petroleum ether with the mass volume of 5 times, degreasing and leaching for 2 hours, centrifuging for 5 minutes at 4 ℃ at 6000r/min, repeatedly leaching and degreasing for three times, centrifuging to obtain solid residues, adding 10% NaOH according to the mass ratio of 1: 5, performing size mixing, adjusting the pH to 9, stirring, centrifuging for 5 minutes at 6000r/min, collecting filtrate, adjusting the pH of the filtrate to 4 with 8% HCl, standing for 1 hour, collecting precipitates, washing the precipitates with deionized water to be neutral, freezing and storing, and drying for 72 hours with a vacuum freeze dryer to obtain proteins for later use; weighing 0.5g of gibberellin and 0.1g of indoleacetic acid, dissolving in 1L of absolute ethanol to prepare a gibberellin-indoleacetic acid ethanol solution, taking 30mL of gibberellin-indoleacetic acid ethanol solution and 5g of protein, uniformly mixing, stirring in the dark for reaction for 2h, and then adding 5mmol/L of CaCl into the mixed solution280mL, standing at room temperature for 16h, desalting with 14000D dialysis bag for 72h, and freeze-drying to obtain gibberellin-indoleacetic acid protein complex.
The liquid culture medium comprises the following components: weighing 25g of bean sprouts, adding 75mL of water, boiling for 20min, filtering and removing residues, wherein the volume is 100 mL; adding 5g glucose, dissolving, mixing, adjusting pH to 7.0, and sterilizing at 121 deg.C for 30 min.
The specific composition of the culture medium in the seeding tank is as follows: 0.3% of beef extract, 1% of peptone, 0.5% of sodium chloride and 0.5% of glucose.
The bacillus subtilis, the bacillus licheniformis, the lactobacillus plantarum, the bacillus megaterium and the bacillus mucilaginosus are all sold in the market, and the effective viable count is 100-200 billion/g.
And (4) during packaging, storing the component B in a vacuum and dark state.
When in use, the component B is dissolved in 10 times of aqueous solution, and the component A is added and mixed evenly for use. When the biological agent is actually used, the biological agent is diluted by 500-800 times and is sprayed for use.
Comparative example 1
A microbial agent capable of promoting flower bud differentiation of plants comprises a component A and a component B, wherein the component A is prepared from the following raw materials in parts by weight: 20 parts of bacillus subtilis, 10 parts of bacillus licheniformis, 10 parts of lactobacillus plantarum, 8 parts of bacillus megaterium, 5 parts of bacillus mucilaginosus and 3 parts of a dispersing agent; the component B is 5-10 parts of gibberellin-indoleacetic acid and is prepared by the following steps:
0.5g of gibberellin and 0.1g of indoleacetic acid are weighed and dissolved in 1L of absolute ethyl alcohol to prepare gibberellin-indoleacetic acid ethanol solution, and then 5mmol/L CaCl is added into the mixed solution280mL, and freeze-drying to obtain the gibberellin-indoleacetic acid.
This comparative example is the same as example 3 except that protein complexation is not performed.
Comparative example 2
A microbial agent capable of promoting flower bud differentiation of plants comprises a component A and a component B, wherein the component A is prepared from the following raw materials in parts by weight: 20 parts of bacillus subtilis, 10 parts of bacillus licheniformis, 10 parts of lactobacillus plantarum, 8 parts of bacillus megaterium and 5 parts of bacillus mucilaginosus; the component B is 5-10 parts of gibberellin-indoleacetic acid-protein complex.
This comparative example is the same as example 3 except that no dispersant is added.
Comparative example 3
A microbial agent capable of promoting flower bud differentiation of plants comprises a component A and a component B, wherein the component A is prepared from the following raw materials in parts by weight: 20 parts of bacillus subtilis, 10 parts of bacillus licheniformis, 10 parts of lactobacillus plantarum, 8 parts of bacillus megaterium and 5 parts of bacillus mucilaginosus; the component B is gibberellin-indoleacetic acid.
The comparative example is the same as example 3 except that protein compounding is not performed and no dispersant is added.
Demonstration of the experiment
Materials and methods
The test site is arranged in a certain vegetable greenhouse of the Weiguan city of Shandong province, the test area is 20 mu, and the soil fertility level is uniform.
The test crop tomato has the row spacing of 50cm by 50cm, and the growth vigor is moderate and consistent. The variety is "greenhouse 68".
The test mulching film is a polyethylene film, the thickness of the film is 0.012mm, and the width of the film is 1.2 m.
The test sites were subjected to 3 test treatments:
experimental groups: applying the fertilizer to be tested once within one week after field planting, diluting by 800 times, and 500 mL/mu;
control group (CK): each mu is carried out with the same amount of clear water in the same experimental group at the same time, and other fertilization measures are the same.
Comparative example group: the fertilizers prepared in comparative examples 1-3 with the same amount as the experimental group are applied to each mu at the same time, and other fertilizing measures are the same.
Detection method
Determination of vitamin C in tomato fruits: weighing about 100g of mixed sample in each of 12-14 fresh fruits, adding 100ml of 2% oxalic acid solution, beating into slurry in a tissue triturator, taking about 20g of slurry, diluting to 100ml with 1% oxalic acid, shaking up, filtering, and measuring by using a 2, 4-dinitrophenylhydrazine colorimetric method.
Determination of reducing sugars in tomato fruits: taking 12-14 fresh fruits, mashing the fruits into pulp, weighing about 10g of the pulp in a 250ml volumetric flask, removing acidity and precipitated protein by using calcium carbonate and 10% lead acetate solution, extracting in a water bath at 80 ℃, and measuring by using a cyanide salt iodine method.
Determination of acidity in tomato fruits: taking 12-14 fresh fruits, mashing the fruits in a mashing machine, weighing 5g of a uniform mashed sample, and then titrating with 0.1mol/L sodium hydroxide solution.
TABLE 1 results of the experiment
It should be noted that the above-mentioned embodiments are only some of the preferred modes for implementing the invention, and not all of them. Obviously, all other embodiments obtained by persons of ordinary skill in the art based on the above-mentioned embodiments of the present invention without any creative effort shall fall within the protection scope of the present invention.
Claims (4)
1. The microbial agent capable of promoting flower bud differentiation of plants is characterized by comprising a component A and a component B, wherein the component A is prepared from the following raw materials in parts by weight: 10-20 parts of bacillus subtilis, 5-10 parts of bacillus licheniformis, 5-10 parts of lactobacillus plantarum, 5-8 parts of bacillus megaterium, 3-5 parts of bacillus mucilaginosus and 1-3 parts of a dispersing agent; the component B is 5-10 parts of gibberellin-indoleacetic acid-protein complex and is prepared by the following steps:
1) drying peanut kernels, removing red skins, crushing, sieving with a 60-mesh sieve, adding petroleum ether with the mass volume of 5 times, degreasing and leaching for 2 hours, centrifuging for 5 minutes at 4 ℃ at 6000r/min, repeatedly leaching and degreasing for three times, centrifuging to obtain solid residues, adding 10% NaOH according to the mass ratio of 1: 5, performing size mixing, adjusting the pH to 9, stirring, centrifuging for 5 minutes at 6000r/min, collecting filtrate, adjusting the pH of the filtrate to 4 with 8% HCl, standing for 1 hour, collecting precipitates, washing the precipitates with deionized water to be neutral, freezing and storing, and drying for 72 hours with a vacuum freeze dryer to obtain proteins for later use;
2) weighing 0.5g of gibberellin and 0.1g of indoleacetic acid, dissolving in 1L of absolute ethanol to prepare a gibberellin-indoleacetic acid ethanol solution, taking 30mL of gibberellin-indoleacetic acid ethanol solution and 5g of protein prepared in the step (1), uniformly mixing, stirring in the dark for reaction for 2h, and then adding 5mmol/L of CaCl into the mixed solution280mL, standing at room temperature for 16h, desalting with 14000D dialysis bag for 72h, and freeze-drying to obtain gibberellin-indoleacetic acid protein complex.
2. The microbial agent capable of promoting flower bud differentiation of plants according to claim 1, comprising a component A and a component B, wherein the component A is prepared from the following raw materials in parts by weight: 20 parts of bacillus subtilis, 10 parts of bacillus licheniformis, 10 parts of lactobacillus plantarum, 8 parts of bacillus megaterium, 5 parts of bacillus mucilaginosus and 3 parts of a dispersing agent; the component B is 10 parts of gibberellin-protein complex.
3. The microbial agent capable of promoting plant flower bud differentiation according to claim 1 or 2, wherein the dispersant is obtained by mixing sodium lignosulfonate and chitosan according to a mass ratio of 1: 1.
4. The microbial agent for promoting flower bud differentiation according to claim 1 or 2, wherein the component B is dissolved in 10 times of aqueous solution, and the component A is added and mixed uniformly.
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