Growth regulator for improving peanut yield and stress resistance and preparation method thereof

Abstract

The present invention relates to a growth regulator for improving peanut yield and stress resistance and a preparation method thereof, and belongs to the technical field of plant growth regulators. The growth regulator of the present invention contains the following ingredients: 0.03-0.05 parts by weight of brassinolide, 0.1-0.12 parts by weight of sodium nitrophenolate, 8-12 parts by weight of sorghum root extract, 10-15 parts by weight of biochar, 4-7 parts by weight of chlorella extract, and 12-15 parts by weight of sodium hyaluronate. The plant growth regulator of the present invention can significantly improve the ability of peanuts to resist salt stress, alkali stress, and saline-alkali stress, and can significantly increase its yield and quality when planted in saline-alkali land.

Description

Technical Field

[0001] The invention belongs to the technical field of plant growth regulators and relates to a growth regulator capable of improving peanut yield and stress resistance and a preparation method thereof. Background Art

[0002] Saline is a type of salt accumulation, which refers to land where the salt contained in the soil affects the normal growth of crops. Saline land can be divided into light saline land, moderate saline land and heavy saline land. The essence of its formation is mainly the horizontal and vertical redistribution of various soluble salts on the ground, so that salt gradually accumulates on the surface of the soil in the salt-collecting area. Soil salinization can inhibit or harm plant growth. The survival rate of crops planted in saline-alkali land is not high, resulting in a decrease in yield and quality. The optimization and rational development and utilization of saline-alkali soil resources are of great significance to improving the ecological environment.

[0003] Peanuts are moderately salt-tolerant crops and important economic and oil crops. Expanding the area of ​​peanut cultivation in saline-alkali land can ease the pressure on my country's edible oil supply and achieve high utilization of saline-alkali soil. However, under saline-alkali stress, peanut plants show reduced root vitality, insufficient growth, and weakened photosynthesis, resulting in reduced pod yield and reduced kernel quality. How to increase peanut resistance and increase yield is a major problem that needs to be solved urgently. Summary of the invention

[0004] The main purpose of the present invention is to provide a plant growth regulator, which can significantly improve the stress resistance of peanuts in a saline-alkali environment and help improve the yield and quality of peanuts planted in saline-alkali land.

[0005] The present invention adopts the following technical solutions to achieve the above purpose:

[0006] A growth regulator for improving peanut yield and stress resistance, comprising the following ingredients: 0.03-0.05 parts by weight of brassinolide, 0.1-0.12 parts by weight of sodium nitrophenolate, 8-12 parts by weight of sorghum root extract, 10-15 parts by weight of biochar, 4-7 parts by weight of chlorella extract, and 12-15 parts by weight of sodium hyaluronate.

[0007] The preparation method of the sorghum root extract of the present invention is:

[0008] The crushed sorghum root is added to a 75% ethanol solution by mass and refluxed for extraction for 1-1.5 hours, residual ethanol is removed, and the solution is cooled to room temperature to obtain a mixed solution; NaCl, yeast extract powder, and tryptone are added to the mixed solution to adjust the pH to 7.0-7.5, and Bacillus cereus seed solution and Halomonas songnensis seed solution are inoculated, and the solution is cultured at 25-30°C for 1.5-2 days, and then lysozyme and β-glucanase are added to perform enzymolysis at 35-40°C for 8-10 hours, and then ultrasonic treatment is performed, and the pH is adjusted to 6.0-6.5, and Bacillus subtilis fibrinolytic enzyme and lumbrokinase are added to perform enzymolysis at 35-40°C for 10-12 hours, and the enzyme is inactivated at high temperature; supernatant I and precipitate I are obtained by centrifugation, supernatant I is concentrated, the sample is loaded onto a silica gel column, eluted with ethyl acetate, the eluate is collected, and the sorghum root extract is obtained by drying;

[0009] Preferably, in the preparation method of the sorghum root extract, the enzyme activity of Bacillus subtilis fibrinolytic enzyme is 3000 U / g, the enzyme activity of lumbrokinase is 10000 U / mg, the enzyme activity of lysozyme is 20000 U / mg, and the enzyme activity of β-glucan is 100000 U / g.

[0010] Preferably, in the preparation method of the sorghum root extract, the added amount of Bacillus subtilis fibrinolytic enzyme is 1.2-1.5 g / L, the added amount of lumbrokinase is 0.5-0.8 g / L, the added amount of lysozyme is 6-8 g / L, and the added amount of β-glucan is 3-5 g / L.

[0011] Preferably, in the method for preparing the sorghum root extract, the inoculation amount of Bacillus cereus seed solution is 1-1.5% of the volume of the mixed solution, and the inoculation amount of Halomonas songnensis seed solution is 3-5% of the volume of the mixed solution.

[0012] Preferably, in the method for preparing the sorghum root extract, the OD value of the Bacillus cereus seed solution is 0.4-0.5, and the OD value of the Halomonas songnensis seed solution is 0.2-0.3.

[0013] Preferably, in the method for preparing the sorghum root extract, the amount of NaCl added is 10-12 g / L, the amount of yeast extract powder added is 10-15 g / L, and the amount of tryptone added is 12-15 g / L.

[0014] The preparation method of the chlorella extract of the present invention is:

[0015] Chlorella is added with water to form a homogenate, the pH is adjusted to 5.0-5.5, pectin lyase and cellobiase are added, and enzymolysis is carried out at 35-40°C for 6-8 hours, and then the pH is adjusted to 6.5-7.0, dextranase, mannanase and neutral protease are added, and enzymolysis is carried out at 35-40°C for 6-8 hours, and the enzyme is inactivated at high temperature to obtain an enzymolysis solution; the pH of the enzymolysis solution is adjusted to 3.5-4.0, and the solution is allowed to stand for 12-15 hours, and centrifuged to obtain a supernatant II and a precipitate II, and the supernatant is dried to obtain a Chlorella extract.

[0016] Preferably, in the preparation method of the above-mentioned Chlorella extract, the enzyme activity of pectin lyase is 600U / mL, the enzyme activity of cellobiase is 1000U / mL, the enzyme activity of dextranase is 25000U / mL, the enzyme activity of mannanase is 50000U / g, and the enzyme activity of neutral protease is 60000U / g.

[0017] Preferably, in the preparation method of the above-mentioned Chlorella extract, the added amount of pectin lyase is 1-1.5 mL / L, the added amount of cellobiase is 0.5-0.8 mL / L, the added amount of dextranase is 1.0-1.2 mL / L, the added amount of mannanase is 1-1.5 g / L, and the added amount of neutral protease is 2-2.3 g / L.

[0018] The preparation method of biochar of the present invention is:

[0019] The precipitate I obtained in the above-mentioned method for preparing the sorghum root extract and the precipitate II obtained in the method for preparing the chlorella extract are mixed to obtain a mixture; the mixture is soaked in a potassium permanganate solution with a mass fraction of 5-8% for 8-10 hours, dried, calcined in an oxygen-free environment at 600-700°C for 3-5 hours, and then wood vinegar and β-nicotinamide mononucleotide are added, mixed and dried to obtain biochar.

[0020] Preferably, in the above-mentioned method for preparing biochar, the amount of precipitate II added is 0.8-1.0 times the mass of precipitate I, the amount of wood vinegar added is 1.2-1.5 times the mass of the mixture, and the amount of β-nicotinamide mononucleotide added is 0.8-1.0 times the mass of the mixture.

[0021] The preparation method of the growth regulator of the present invention is:

[0022] Sodium hyaluronate is dissolved in water, biochar is added and mixed, and then dried. Brassinolide, sodium nitrophenolate, sorghum root extract and chlorella extract are added and mixed to obtain a growth regulator.

[0023] The present invention has the following beneficial effects:

[0024] 1. The plant growth regulator of the present invention is added with sorghum root extract fermented by salt-alkali tolerant microorganisms. During the fermentation process, the microorganisms grow and reproduce in a saline-alkali environment using sorghum root, yeast extract and other ingredients, and produce extracellular metabolites during their growth and reproduction. Under the action of lysozyme and saccharolytic enzymes, the bacteria rupture and release intracellular metabolites. After the sorghum root is fermented by microorganisms, the macromolecular substances in it are converted into small molecular products; the fermented product is further decomposed after enzymatic hydrolysis, and then separated and purified to obtain sorghum root extract. The components in the sorghum root extract can significantly improve the stress resistance of peanuts. The regulator of the present invention also adds chlorella extract. During its preparation process, a two-step enzymatic hydrolysis method is adopted to utilize the decomposition action of different enzymes to decompose the components in chlorella to the maximum extent, and then the pH is adjusted to remove some ineffective components to obtain the chlorella extract that can improve the stress resistance of peanuts.

[0025] 2. The present invention further utilizes the extracted sorghum root and chlorella, mixes them and prepares biochar, utilizes the adsorption performance of the rich pores of biochar, effectively loads wood vinegar, β-nicotinamide mononucleotide, and sodium hyaluronate therein, and when used for peanut planting, wood vinegar and β-nicotinamide mononucleotide can be slowly released to maintain the stress resistance of peanuts in saline-alkali environments. At the same time, hyaluronic acid has high permeability and biocompatibility, and it can carry small molecules of sorghum root extract and chlorella extract into peanut tissues, activate cell activity, and improve the stress resistance of peanuts.

[0026] 3. The salt-alkali stress experiment proved that the synergistic effect between the components of the plant growth regulator of the present invention can significantly improve the seed germination rate of peanuts under salt stress, alkali stress and salt-alkali stress, and promote plant growth; in the field experiment, the plant growth regulator of the present invention can significantly increase the yield of peanuts in saline-alkali land, and the peanut fruits are plump and of high quality. DETAILED DESCRIPTION

[0027] The present invention is further explained below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not used to limit the scope of the present invention. After reading the present invention, modifications of various equivalent forms of the present invention by those skilled in the art all fall within the scope of protection of the claims of this application. The microorganisms and enzymes used in the present invention are all existing products and can be purchased from microbial preservation centers or online and offline channels.

[0028] Example 1

[0029] 1. Preparation of Sorghum Root Extract

[0030] 1.1 Seed solution preparation:

[0031] The Bacillus cereus strain with the deposit number of CGMCC 1.9067 purchased from the deposit center was activated according to the instructions, inoculated into the seed culture medium and cultured until the OD value was 0.5 to obtain the Bacillus cereus seed liquid;

[0032] The Halomonas songnensis strain with the deposit number of CGMCC 1.12152 purchased from the deposit center was activated according to the instructions, inoculated into the seed culture medium and cultured until the OD value was 0.2 to obtain the Halomonas songnensis seed solution.

[0033] 1.2 Preparation of sorghum root extract:

[0034] The discarded sorghum roots were washed, air-dried, and crushed; the crushed sorghum roots were added to a 75% ethanol solution and refluxed for extraction for 1 hour to remove the residual ethanol, and the solution was cooled to room temperature to obtain a mixed solution; 12 g / L NaCl, 10 g / L yeast extract, and 15 g / L tryptone were added to the mixed solution, the pH was adjusted to 7.5, and the above-mentioned Bacillus cereus seed solution (inoculation amount was 1.5% of the volume of the mixed solution) and the songnensis seed solution (inoculation amount was 3% of the volume of the mixed solution) were inoculated, and cultured at 30°C for 1.5 days, and then 8 g / L of solution was added. The extract was enzymatically treated with 3 g / L beta-glucanase (enzyme activity of 20,000 U / mg) and 3 g / L beta-glucanase (enzyme activity of 100,000 U / g) at 35°C for 10 h, then ultrasonically treated, and the pH was adjusted to 6.5. 1.5 g / L Bacillus subtilis fibrinolytic enzyme (enzyme activity of 3,000 U / g) and 0.5 g / L lumbrokinase (enzyme activity of 10,000 U / mg) were added, and the extract was enzymatically treated at 35°C for 12 h, and the enzyme was inactivated by high temperature. The supernatant I and precipitate I were obtained by centrifugation. The supernatant I was concentrated, loaded onto a silica gel column, eluted with ethyl acetate, and the eluate was collected and dried to obtain the sorghum root extract.

[0035] 2. Preparation of Chlorella Extract:

[0036] Chlorella was homogenized by adding water, the pH was adjusted to 5.5, 1 mL / L pectin lyase (enzyme activity of 600 U / mL) and 0.8 mL / L cellobiase (enzyme activity of 1000 U / mL) were added, and enzymolysis was carried out at 40°C for 6 hours. The pH was then adjusted to 6.5, 1.2 mL / L dextranase (enzyme activity of 25000 U / mL), 1 g / L mannanase (enzyme activity of 50000 U / g) and 2.3 g / L neutral protease (enzyme activity of 60000 U / g) were added, and enzymolysis was carried out at 40°C for 6 hours. The enzyme was inactivated at high temperature to obtain an enzymolysis solution. The pH of the enzymolysis solution was adjusted to 3.5, the solution was allowed to stand for 12 hours, and the solution was centrifuged to obtain supernatant II and precipitate II. The supernatant was dried to obtain a Chlorella extract.

[0037] 3. Preparation of biochar:

[0038] The precipitate I obtained in the preparation of the sorghum root extract and the precipitate II obtained in the preparation of the chlorella extract were mixed in a mass ratio of 1:0.8 to obtain a mixture; the mixture was immersed in a potassium permanganate solution with a mass fraction of 8% for 8 hours, dried, and calcined in an oxygen-free environment at 700°C for 3 hours, and then 1.5 times the mass of wood vinegar and 0.8 times the mass of β-nicotinamide mononucleotide were added, mixed and dried to obtain biochar.

[0039] 4. Preparation of growth regulators:

[0040] 13 parts of sodium hyaluronate are dissolved in water, 13 parts of biochar are added and mixed, then dried, 0.04 parts of brassinolide, 0.11 parts of sodium nitrophenolate, 10 parts of sorghum root extract, and 6 parts of chlorella extract are added and mixed to obtain a growth regulator. The 1 part mentioned in the present invention can be a conventional weight unit such as 1g or 1kg.

[0041] Example 2

[0042] 1. Preparation of Sorghum Root Extract

[0043] 1.1 Seed solution preparation:

[0044] The Bacillus cereus strain with the deposit number of CGMCC 1.9067 purchased from the deposit center was activated according to the instructions, inoculated into the seed culture medium and cultured until the OD value was 0.4 to obtain the Bacillus cereus seed solution;

[0045] The Halomonas songnensis strain with the deposit number of CGMCC 1.12152 purchased from the deposit center was activated according to the instructions, inoculated into the seed culture medium and cultured until the OD value was 0.3 to obtain the Halomonas songnensis seed solution.

[0046] 1.2 Preparation of sorghum root extract:

[0047] The discarded sorghum roots were washed, air-dried, and crushed; the crushed sorghum roots were added to a 75% ethanol solution and refluxed for extraction for 1.5 h to remove the residual ethanol, and the solution was cooled to room temperature to obtain a mixed solution; 10 g / L NaCl, 15 g / L yeast extract, and 12 g / L tryptone were added to the mixed solution, the pH was adjusted to 7.0, and the above-mentioned Bacillus cereus seed solution (inoculation amount was 1% of the volume of the mixed solution) and the songnensis seed solution (inoculation amount was 5% of the volume of the mixed solution) were inoculated, and cultured at 25°C for 2 days, and then 6 g / L of solution was added. The enzyme (enzyme activity is 20000U / mg) and 5g / L β-glucanase (enzyme activity is 100000U / g) were enzymolyzed at 40℃ for 8h, then ultrasonicated, and the pH was adjusted to 6.0, 1.2g / L Bacillus subtilis fibrinolytic enzyme (enzyme activity is 3000U / g) and 0.8g / L lumbrokinase (enzyme activity is 10000U / mg) were added, and the enzyme was enzymolyzed at 40℃ for 10h, and the enzyme was inactivated at high temperature; the supernatant I and precipitate I were obtained by centrifugation, the supernatant I was concentrated, the sample was loaded on a silica gel column, eluted with ethyl acetate, the eluate was collected, and dried to obtain the sorghum root extract;

[0048] 2. Preparation of Chlorella Extract:

[0049] Chlorella was homogenized by adding water, the pH was adjusted to 5.0, 1.5 mL / L pectin lyase (enzyme activity of 600 U / mL) and 0.5 mL / L cellobiase (enzyme activity of 1000 U / mL) were added, and enzymolysis was performed at 35°C for 8 hours, and then the pH was adjusted to 7.0, 1.0 mL / L dextranase (enzyme activity of 25000 U / mL), 1.5 g / L mannanase (enzyme activity of 50000 U / g) and 2 g / L neutral protease (enzyme activity of 60000 U / g) were added, and enzymolysis was performed at 35°C for 8 hours, and the enzyme was inactivated at high temperature to obtain an enzymolysis solution; the pH of the enzymolysis solution was adjusted to 4.0, and the solution was allowed to stand for 15 hours, and centrifuged to obtain supernatant II and precipitate II, and the supernatant was dried to obtain a Chlorella extract.

[0050] 3. Preparation of biochar:

[0051] The precipitate I obtained in the preparation of the sorghum root extract and the precipitate II obtained in the preparation of the chlorella extract were mixed in a mass ratio of 1:1.0 to obtain a mixture; the mixture was immersed in a 5% potassium permanganate solution for 10 hours, dried, and calcined in an oxygen-free environment at 600°C for 5 hours, and then 1.2 times the mass of wood vinegar and 1.0 times the mass of β-nicotinamide mononucleotide were added, mixed and dried to obtain biochar.

[0052] 4. Preparation of growth regulators:

[0053] 12 parts of sodium hyaluronate are dissolved in water, 15 parts of biochar are added and mixed, then dried, 0.05 parts of brassinolide, 0.1 parts of sodium nitrophenolate, 12 parts of sorghum root extract, and 4 parts of chlorella extract are added and mixed to obtain a growth regulator. The 1 part mentioned in the present invention can be a conventional weight unit such as 1g or 1kg.

[0054] Example 3

[0055] 1. Preparation of Sorghum Root Extract

[0056] 1.1 Seed solution preparation:

[0057] The Bacillus cereus strain with the deposit number of CGMCC 1.9067 purchased from the deposit center was activated according to the instructions, inoculated into the seed culture medium and cultured until the OD value was 0.5 to obtain the Bacillus cereus seed liquid;

[0058] The Halomonas songnensis strain with the deposit number of CGMCC 1.12152 purchased from the deposit center was activated according to the instructions, inoculated into the seed culture medium and cultured until the OD value was 0.3 to obtain the Halomonas songnensis seed solution.

[0059] 1.2 Preparation of sorghum root extract:

[0060] The discarded sorghum roots were washed, air-dried, and crushed; the crushed sorghum roots were added to a 75% ethanol solution and refluxed for extraction for 1.5 h, residual ethanol was removed, and the solution was cooled to room temperature to obtain a mixed solution; 11 g / L NaCl, 13 g / L yeast extract, and 13 g / L tryptone were added to the mixed solution, the pH was adjusted to 7.5, and the above-mentioned Bacillus cereus seed solution (inoculation amount was 1.3% of the volume of the mixed solution) and Songnenha Halomonas seed solution (inoculation amount was 4% of the volume of the mixed solution) were inoculated, and cultured at 30°C for 1.5 days, and then 7 g / L Lysozyme (enzyme activity is 20000U / mg) and 4g / L β-glucanase (enzyme activity is 100000U / g) were enzymolyzed at 35℃ for 10h, then ultrasonicated, and the pH was adjusted to 6.0, 1.3g / L Bacillus subtilis fibrinolytic enzyme (enzyme activity is 3000U / g) and 0.6g / L lumbrokinase (enzyme activity is 10000U / mg) were added, and enzymolyzed at 40℃ for 10h, and the enzyme was inactivated by high temperature; supernatant I and precipitate I were obtained by centrifugation, supernatant I was concentrated, loaded on a silica gel column, eluted with ethyl acetate, the eluate was collected, and dried to obtain sorghum root extract;

[0061] 2. Preparation of Chlorella Extract:

[0062] Chlorella was homogenized by adding water, the pH was adjusted to 5.5, 1.3 mL / L pectin lyase (enzyme activity of 600 U / mL) and 0.7 mL / L cellobiase (enzyme activity of 1000 U / mL) were added, and enzymolysis was performed at 40°C for 6 hours, then the pH was adjusted to 7.0, 1.1 mL / L dextranase (enzyme activity of 25000 U / mL), 1.3 g / L mannanase (enzyme activity of 50000 U / g) and 2.2 g / L neutral protease (enzyme activity of 60000 U / g) were added, and enzymolysis was performed at 40°C for 8 hours, and the enzyme was inactivated at high temperature to obtain an enzymolysis solution; the pH of the enzymolysis solution was adjusted to 4.0, the solution was allowed to stand for 13 hours, and centrifuged to obtain supernatant II and precipitate II, and the supernatant was dried to obtain a Chlorella extract.

[0063] 3. Preparation of biochar:

[0064] The precipitate I obtained in the preparation of the sorghum root extract and the precipitate II obtained in the preparation of the chlorella extract were mixed in a mass ratio of 1:0.9 to obtain a mixture; the mixture was immersed in a 7% potassium permanganate solution for 9 hours, dried, and calcined in an oxygen-free environment at 700°C for 4 hours, and then 1.3 times the mass of wood vinegar and 0.9 times the mass of β-nicotinamide mononucleotide were added, mixed and dried to obtain biochar.

[0065] 4. Preparation of growth regulators:

[0066] 15 parts of sodium hyaluronate are dissolved in water, 10 parts of biochar are added and mixed, then dried, 0.03 parts of brassinolide, 0.12 parts of sodium nitrophenolate, 8 parts of sorghum root extract, and 7 parts of chlorella extract are added and mixed to obtain a growth regulator. The 1 part mentioned in the present invention can be a conventional weight unit such as 1g or 1kg.

[0067] Comparative Example 1

[0068] 1. Preparation of Sorghum Root Extract

[0069] The discarded sorghum roots were washed, air-dried, and crushed; the crushed sorghum roots were added to a 75% ethanol solution for reflux extraction for 1.5 hours, residual ethanol was removed, and the mixture was cooled to room temperature to obtain a mixed solution; the pH of the mixed solution was adjusted to 6.0, 1.3 g / L Bacillus subtilis fibrinolytic enzyme (enzyme activity of 3000 U / g) and 0.6 g / L lumbrokinase (enzyme activity of 10000 U / mg) were added, and the mixture was enzymatically hydrolyzed at 40°C for 10 hours, and the enzyme was inactivated at high temperature; the supernatant and precipitate were obtained by centrifugation, the supernatant I was concentrated, the sample was loaded on a silica gel column, eluted with ethyl acetate, the eluate was collected, and the sorghum root extract was obtained by drying;

[0070] 2. Preparation of biochar:

[0071] The precipitate obtained in the preparation of the sorghum root extract was soaked in a 7% potassium permanganate solution for 9 hours, dried, and calcined in an oxygen-free environment at 700°C for 4 hours. Then, 1.3 times the mass of wood vinegar and 0.9 times the mass of β-nicotinamide mononucleotide were added, mixed and dried to obtain biochar.

[0072] 3. Preparation of growth regulators:

[0073] 15 parts of sodium hyaluronate are dissolved in water, 10 parts of biochar are added and mixed, then dried, 0.03 parts of brassinolide, 0.12 parts of sodium nitrophenolate and 8 parts of sorghum root extract are added and mixed to obtain a growth regulator. 1 part in the present invention can be a conventional weight unit such as 1g or 1kg.

[0074] Comparative Example 2

[0075] 1. Preparation of Sorghum Root Extract

[0076] 1.1 Seed solution preparation:

[0077] The Bacillus cereus strain with the deposit number of CGMCC 1.9067 purchased from the deposit center was activated according to the instructions, inoculated into the seed culture medium and cultured until the OD value was 0.5 to obtain the Bacillus cereus seed liquid;

[0078] The Halomonas songnensis strain with the deposit number of CGMCC 1.12152 purchased from the deposit center was activated according to the instructions, inoculated into the seed culture medium and cultured until the OD value was 0.3 to obtain the Halomonas songnensis seed solution.

[0079] 1.2 Preparation of sorghum root extract:

[0080] The discarded sorghum root is taken, cleaned, air-dried, crushed, and water is added to prepare a suspension; 11 g / L NaCl, 13 g / L yeast extract, and 13 g / L tryptone are added to the suspension, the pH is adjusted to 7.5, the above-mentioned Bacillus cereus seed solution (the inoculation amount is 1.3% of the volume of the mixed solution) and the Songnenha Halomonas seed solution (the inoculation amount is 4% of the volume of the mixed solution) are inoculated, and cultured at 30° C. for 1.5 days, and then 7 g / L lysozyme (enzyme activity is 20000 U / mg) is added to perform enzymolysis at 35° C. for 10 hours, the enzyme is inactivated by high temperature, and supernatant I is obtained by centrifugation, and the supernatant I is dried to obtain a sorghum root extract;

[0081] 2. Preparation of Chlorella Extract:

[0082] Chlorella was added with water to prepare a homogenate, the pH was adjusted to 5.5, 1.3 mL / L pectin lyase (enzyme activity was 600 U / mL) and 0.7 mL / L cellobiase (enzyme activity was 1000 U / mL) were added, and enzymolysis was performed at 40°C for 6 hours, and the enzyme was inactivated at high temperature to obtain an enzymolysis solution; the pH of the enzymolysis solution was adjusted to 4.0, and the solution was allowed to stand for 13 hours, and centrifuged to obtain supernatant II, and the supernatant was dried to obtain the Chlorella extract.

[0083] 3. Preparation of growth regulators:

[0084] Mix 15 parts of sodium hyaluronate, 0.03 parts of brassinolide, 0.12 parts of sodium nitrophenolate, 8 parts of sorghum root extract, and 7 parts of chlorella extract to obtain a growth regulator. The 1 part mentioned in the present invention can be a conventional weight unit such as 1g or 1kg.

[0085] Comparative Example 3

[0086] 1. Preparation of Sorghum Root Extract

[0087] 1.1 Seed solution preparation:

[0088] The Bacillus cereus strain with the deposit number of CGMCC 1.9067 purchased from the deposit center was activated according to the instructions, inoculated into the seed culture medium and cultured until the OD value was 0.5 to obtain the Bacillus cereus seed liquid;

[0089] 1.2 Preparation of sorghum root extract:

[0090] The discarded sorghum roots were cleaned, air-dried, and crushed; the crushed sorghum roots were added to a 75% ethanol solution and refluxed for extraction for 1.5 hours, residual ethanol was removed, and the mixture was cooled to room temperature to obtain a mixed solution; the mixed solution was inoculated with the above-mentioned Bacillus cereus seed solution (the inoculation amount was 1.3% of the volume of the mixed solution), and cultured at 30°C for 1.5 days, and then 7g / L lysozyme (enzyme activity was 20000U / mg) and 4g / L β-glucanase (enzyme activity was 100000U / g) were added for enzymolysis at 35°C for 10 hours, and the enzyme was inactivated at high temperature; supernatant I and precipitate were obtained by centrifugation, and the supernatant I was dried to obtain the sorghum root extract;

[0091] 2. Preparation of Chlorella Extract:

[0092] Chlorella is added with water to make a homogenate, the pH is adjusted to 5.5, 1.3 mL / L pectin lyase (enzyme activity is 600 U / mL), 1.3 g / L mannanase (enzyme activity is 50000 U / g), and 2.2 g / L neutral protease (enzyme activity is 60000 U / g) are added, and enzymolysis is carried out at 40°C for 8 hours, and the enzyme is inactivated at high temperature to obtain an enzymolysis solution; the pH of the enzymolysis solution is adjusted to 4.0, and the solution is allowed to stand for 13 hours, and centrifuged to obtain supernatant II, and the supernatant II is dried to obtain the Chlorella extract.

[0093] 3. Preparation of biochar:

[0094] The precipitate I obtained in the preparation of the sorghum root extract was calcined at 700°C for 4 hours, and then 1.3 times the mass of wood vinegar was added, mixed and dried to obtain biomass charcoal.

[0095] 4. Preparation of growth regulators:

[0096] Mix 10 parts of biochar, 0.03 parts of brassinolide, 0.12 parts of sodium nitrophenolate, 8 parts of sorghum root extract, and 7 parts of chlorella extract to obtain a growth regulator. The 1 part mentioned in the present invention can be a conventional weight unit such as 1g or 1kg.

[0097] Performance Testing

[0098] 1. Effects of growth regulators on peanut seed germination under saline-alkali stress

[0099] 1. Experimental Procedure

[0100] 1.1 Salt stress experiment: Select peanut seeds of uniform size, complete appearance, and fullness, soak them in 10% hydrogen peroxide solution for 5 minutes for disinfection, rinse them with distilled water, absorb the water, and place them in culture dishes numbered Y1-Y8, 50 seeds in each culture dish, and culture them in an incubator at 25±2℃, alternating day and night for 12 hours until the seedling stage. The peanut seeds in the culture dishes numbered Y1-Y7 were treated with salt stress with 0.12mol / L NaCl solution, and the culture dish numbered Y8 was cultured normally with distilled water as a blank. During the culture period, the growth regulator (10 g / L) obtained in Example 1 was sprayed on the peanut seeds in the culture dish No. Y1, the growth regulator (10 g / L) obtained in Example 2 was sprayed on the peanut seeds in the culture dish No. Y2, the growth regulator (10 g / L) obtained in Example 2 was sprayed on the peanut seeds in the culture dish No. Y3, the growth regulator (10 g / L) obtained in Example 3 was sprayed on the peanut seeds in the culture dish No. Y4, the growth regulator (10 g / L) obtained in Comparative Example 1 was sprayed on the peanut seeds in the culture dish No. Y5, the growth regulator (10 g / L) obtained in Comparative Example 2 was sprayed on the peanut seeds in the culture dish No. Y6, and the growth regulator (10 g / L) obtained in Comparative Example 3 was sprayed once every 5 days; the peanut seeds in the culture dish No. Y7 were sprayed with distilled water as a control. Distilled water was regularly supplemented during the culture period to keep the culture environment moist.

[0101] Ten peanut seedlings were randomly selected from each culture dish, and the taproot length, fresh weight and dry weight of aboveground and underground parts were determined, and the activities of peroxidase (POD) and superoxide dismutase (SOD) in the root tip were measured.

[0102] 1.2 Alkali stress test: Select peanut seeds of uniform size, complete appearance and fullness, soak them in 10% hydrogen peroxide solution for 5 minutes for disinfection, rinse them with distilled water, absorb the water, and place them in culture dishes numbered J1-J8, 50 seeds in each culture dish, and culture them in an incubator at 25±2℃ with 12h alternating day and night until the seedling stage. The peanut seeds in the culture dishes numbered J1-J7 were treated with 0.1mol / L NaHCO 3 (pH8.5) solution was used for alkaline stress treatment, and the culture dish No. J8 was cultured normally with distilled water as a blank. During the culture period, the peanut seeds in the culture dish No. J1 were sprayed with the growth regulator (10 g / L) obtained in Example 1, the peanut seeds in the culture dish No. J2 were sprayed with the growth regulator (10 g / L) obtained in Example 2, the peanut seeds in the culture dish No. J3 were sprayed with the growth regulator (10 g / L) obtained in Example 3, the peanut seeds in the culture dish No. J4 were sprayed with the growth regulator (10 g / L) obtained in Comparative Example 1, the peanut seeds in the culture dish No. J5 were sprayed with the growth regulator (10 g / L) obtained in Comparative Example 2, and the peanut seeds in the culture dish No. J6 were sprayed with the growth regulator (10 g / L) obtained in Comparative Example 3, once every 5 days; the peanut seeds in the culture dish No. J7 were sprayed with distilled water as a control. Distilled water was replenished regularly during the culture period to keep the culture environment moist.

[0103] Ten peanut seedlings were randomly selected from each culture dish, and the taproot length, fresh weight and dry weight of aboveground and underground parts were determined, and the activities of peroxidase (POD) and superoxide dismutase (SOD) in the root tip were measured.

[0104] 1.3 Salt-alkali stress test: peanut seeds of uniform size, complete appearance and fullness were selected, soaked in 10% hydrogen peroxide solution for 5 min for disinfection, rinsed with distilled water, and dried. They were placed in culture dishes numbered YJ1-YJ8, with 50 seeds in each culture dish, and cultured in an incubator at 25±2℃ with 12h alternating day and night until the seedling stage. The peanut seeds in the culture dishes numbered YJ1-YJ7 were treated with 0.12mol / LNaCl+0.1mol / L NaHCO 3Solution (adjusting pH 8.5) was subjected to saline-alkali stress treatment, and the culture dish No. YJ8 was normally cultured with distilled water as a blank. During the culture period, the peanut seeds in the culture dish No. YJ1 were sprayed with the growth regulator (10 g / L) obtained in Example 1, the peanut seeds in the culture dish No. YJ2 were sprayed with the growth regulator (10 g / L) obtained in Example 2, the peanut seeds in the culture dish No. YJ3 were sprayed with the growth regulator (10 g / L) obtained in Example 3, the peanut seeds in the culture dish No. YJ4 were sprayed with the growth regulator (10 g / L) obtained in Comparative Example 1, the peanut seeds in the culture dish No. YJ5 were sprayed with the growth regulator (10 g / L) obtained in Comparative Example 2, and the peanut seeds in the culture dish No. YJ6 were sprayed with the growth regulator (10 g / L) obtained in Comparative Example 3, and the spraying was performed once every 5 days; the peanut seeds in the culture dish No. YJ7 were sprayed with distilled water as a control. During the culture period, distilled water was replenished regularly to keep the culture environment moist.

[0105] Ten peanut seedlings were randomly selected from each culture dish, and the taproot length, fresh weight and dry weight of aboveground and underground parts were measured.

[0106] 2. Results and Analysis

[0107] From the results in Table 1, Table 2 and Table 3, it can be seen that the peanut seeds using the plant growth regulator of the present invention have a higher germination rate, better root development and good plant growth under salt stress, alkali stress and saline-alkali stress. Compared with the peanut seeds using the growth regulators of Comparative Example 1, Comparative Example 2 and Comparative Example 3, the germination rate and seedling traits are better.

[0108] Table 1 Characteristics of peanut seedlings under salt stress

[0109]

[0110] Table 2 Characteristics of peanut seedlings under alkaline stress

[0111]

[0112] Table 3 Characteristics of peanut seedlings under saline-alkali stress

[0113]

[0114] 2. Field Experiment

[0115] 2.1 Experimental process

[0116] The experiment was conducted at the cooperative agricultural experimental demonstration base. The peanut variety was "Luhua No. 14". The basic properties of the soil were: organic matter 7.1g / kg, alkaline nitrogen 40.6mg / kg, available phosphorus 30.9mg / kg, available potassium 139.2mg / kg, pH 8.3, and salt content (m / m) 0.31%. Base fertilizer was applied before sowing.

[0117] Divide the peanut seeds into 7 groups, moisten them separately, and mix them in the following way:

[0118] Group 1: commercially available imidacloprid seed coating agent + commercially available fluzolin-methyl seed coating agent + growth regulator of Example 1;

[0119] Group 2: commercially available imidacloprid seed coating agent + commercially available fluazifop-butyl seed coating agent + growth regulator of Example 2;

[0120] Group 3: commercially available imidacloprid seed coating agent + commercially available fluzolin-methyl seed coating agent + growth regulator of Example 3;

[0121] Group 4: commercially available imidacloprid seed coating agent + commercially available fluzolin-methyl seed coating agent + growth regulator of comparative example 1;

[0122] Group 5: commercially available imidacloprid seed coating agent + commercially available fluzolin-methyl seed coating agent + growth regulator of comparative example 2;

[0123] Group 6: commercially available imidacloprid seed coating agent + commercially available fluzolin-methyl seed coating agent + growth regulator of comparative example 3;

[0124] Group 7: commercially available imidacloprid seed coating agent + commercially available fluazifop-butyl seed coating agent.

[0125] The usage amount of imidacloprid seed dressing agent is 40g / 15kg peanut seeds; the usage amount of fluazifop-butyl seed dressing agent is 30g / 15kg peanut seeds; the usage amount of growth regulator is 3g / 15kg peanut seeds.

[0126] In this experiment, peanuts were cultivated with ridge film covering, with a ridge spacing of 90 cm, two rows in one ridge, a row spacing of 30 cm in each ridge, a hole spacing of 20 cm, two peanut seeds in each hole, and a ridge length of 10 m. Two ridges formed a group, and a total of 7 groups were set up, corresponding to the intervals between the groups of peanut seeds after sowing the above-mentioned seed dressing. Among them, groups 1-6 of peanuts were fertilized with corresponding growth regulators during the 5-8 cotyledon stage and the pod stage, and group 7 was only fertilized. The rest of the general field management was the same.

[0127] 2.2 Test indicators

[0128] 2.2.1 During the peanut germination period, observe and record the number of emerged seedlings and calculate the germination rate.

[0129] Germination rate = number of germinated seeds / total number of sown seeds × 100%

[0130] 2.2.2 Peanut traits: At harvest, 20 peanut plants were randomly selected from each group, and the number of fruits per plant and the weight of 100 fruits were calculated.

[0131] 2.2.3 Peanut pod yield: Each group harvested two rows of peanuts with a length of 5 m, and weighed them after natural air drying to calculate the pod yield.

[0132] 3. Results and Analysis

[0133] From the results in Table 4, it can be seen that in the saline-alkali land field experiment, the germination rate of peanuts in Group 1, Group 2, and Group 3 using the plant growth regulator of the present invention is higher, the number of fruits per plant is larger, the fruits are fuller, and the yield is higher. The overall condition is better than the peanuts using the growth regulators of Comparative Examples 1 to Comparative Examples 3.

[0134] Table 4 Characteristics of peanut seedlings under saline-alkali stress

[0135]

[0136] From the above salt-alkali stress test and saline-alkali field experiment results, it can be seen that the synergistic effect between the components in the plant growth regulator provided by the present invention can significantly improve the germination rate of peanut seeds in a saline-alkali environment and improve the salt-alkali resistance of the plant. The growth regulator prepared in Comparative Example 1 does not contain chlorella extract and sorghum root extract, and the preparation method is different. In Comparative Example 2, the preparation methods of sorghum root extract and chlorella extract are different, and there is no biomass charcoal. In Comparative Example 3, the preparation methods of sorghum root extract, chlorella extract, and biomass charcoal are different, and the regulator does not contain sodium hyaluronate. Although the peanut germination rate of the growth regulator obtained by Comparative Examples 1-Comparative Examples 3 is higher than that of the peanut seeds without the regulator, the plant traits, and the yield in the field experiment are increased, but the overall effect is obviously not as good as the peanut seeds using the growth regulator of the present invention.

Claims

1. A growth regulator for improving peanut yield and stress resistance, characterized in that: The growth regulator contains the following ingredients: 0.03-0.05 parts by weight of brassinolide, 0.1-0.12 parts by weight of sodium nitrophenolate, 8-12 parts by weight of sorghum root extract, 10-15 parts by weight of biochar, 4-7 parts by weight of chlorella extract, and 12-15 parts by weight of sodium hyaluronate; The preparation method of the sorghum root extract is: The sorghum root is added to a 75% ethanol solution and refluxed for extraction for 1-1.5 hours, residual ethanol is removed, and the solution is cooled to room temperature to obtain a mixed solution; NaCl, yeast extract powder, and tryptone are added to the mixed solution to adjust the pH to 7.0-7.5, and Bacillus cereus seed solution and Halomonas songnensis seed solution are inoculated, and the solution is cultured at 25-30°C for 1.5-2 days, and then lysozyme and β-glucanase are added to perform enzymolysis at 35-40°C for 8-10 hours, and then ultrasonic treatment is performed, and the pH is adjusted to 6.0-6.5, and Bacillus subtilis fibrinolytic enzyme and lumbrokinase are added to perform enzymolysis at 35-40°C for 10-12 hours, and the enzyme is inactivated at high temperature; supernatant I and precipitate I are obtained by centrifugation, supernatant I is concentrated, the sample is loaded onto a silica gel column, eluted with ethyl acetate, the eluate is collected, and the sorghum root extract is obtained by drying; The preparation method of the chlorella extract is: Chlorella is added with water to form a homogenate, the pH is adjusted to 5.0-5.5, pectin lyase and cellobiase are added, and enzymolysis is carried out at 35-40°C for 6-8 hours, and then the pH is adjusted to 6.5-7.0, dextranase, mannanase and neutral protease are added, and enzymolysis is carried out at 35-40°C for 6-8 hours, and the enzyme is inactivated at high temperature to obtain an enzymolysis solution; the pH of the enzymolysis solution is adjusted to 3.5-4.0, and the solution is allowed to stand for 12-15 hours, and centrifuged to obtain a supernatant II and a precipitate II, and the supernatant II is dried to obtain a Chlorella extract; The preparation method of the biochar is: The precipitate I obtained in the preparation method of sorghum root extract and the precipitate II obtained in the preparation method of chlorella extract are mixed to obtain a mixture; the mixture is soaked in a potassium permanganate solution with a mass fraction of 5-8% for 8-10 hours, dried, calcined in an oxygen-free environment at 600-700°C for 3-5 hours, and then wood vinegar and β-nicotinamide mononucleotide are added, mixed and dried to obtain biochar.

2. The growth regulator according to claim 1, characterized in that In the preparation method of the sorghum root extract, the enzyme activity of Bacillus subtilis fibrinolytic enzyme is 3000U / g, the enzyme activity of lumbrokinase is 10000U / mg, the enzyme activity of lysozyme is 20000U / mg, and the enzyme activity of beta-glucan is 100000U / g.

3. The growth regulator according to claim 1, characterized in that In the preparation method of the sorghum root extract, the added amount of Bacillus subtilis fibrinolytic enzyme is 1.2-1.5 g / L, the added amount of lumbrokinase is 0.5-0.8 g / L, the added amount of lysozyme is 6-8 g / L, and the added amount of beta-glucan is 3-5 g / L.

4. The growth regulator according to claim 1, characterized in that In the preparation method of the sorghum root extract, the inoculation amount of Bacillus cereus seed solution is 1-1.5% of the volume of the mixed solution, and the inoculation amount of Halomonas songnensis seed solution is 3-5% of the volume of the mixed solution.

5. The growth regulator according to claim 1, characterized in that In the preparation method of the sorghum root extract, the OD value of the Bacillus cereus seed solution is 0.4-0.5, and the OD value of the Halomonas songnensis seed solution is 0.2-0.

3.

6. The growth regulator according to claim 1, characterized in that In the preparation method of the sorghum root extract, the added amount of NaCl is 10-12 g / L, the added amount of yeast extract powder is 10-15 g / L, and the added amount of tryptone is 12-15 g / L.

7. The growth regulator according to claim 1, characterized in that In the preparation method of the chlorella extract, the enzyme activity of pectin lyase is 600 U / mL, the enzyme activity of cellobiase is 1000 U / mL, the enzyme activity of dextranase is 25000 U / mL, the enzyme activity of mannanase is 50000 U / g, and the enzyme activity of neutral protease is 60000 U / g.

8. The growth regulator according to claim 1, characterized in that In the preparation method of the Chlorella extract, the added amount of pectin lyase is 1-1.5 mL / L, the added amount of cellobiase is 0.5-0.8 mL / L, the added amount of dextranase is 1.0-1.2 mL / L, the added amount of mannanase is 1-1.5 g / L, and the added amount of neutral protease is 2-2.3 g / L.

9. The growth regulator according to claim 1, characterized in that In the biochar preparation method, the amount of precipitate II added is 0.8-1.0 times the mass of precipitate I, the amount of wood vinegar added is 1.2-1.5 times the mass of the mixture, and the amount of β-nicotinamide mononucleotide added is 0.8-1.0 times the mass of the mixture.

10. The growth regulator according to claim 1, characterized in that The preparation method of the growth regulator is: Sodium hyaluronate is dissolved in water, biochar is added and mixed, and then dried. Brassinolide, sodium nitrophenolate, sorghum root extract and chlorella extract are added and mixed to obtain a growth regulator.

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