Plant growth regulator and preparation method thereof

Abstract

The present invention provides a plant growth regulator and a preparation method thereof, belonging to the technical field of plant growth regulators. After corn stalks, buckwheat seeds and rapeseed pollen are mixed, yeast, Bacillus subtilis and Monascus purpureus are inoculated into the enzymatic hydrolysis product obtained by complex enzyme hydrolysis for fermentation to obtain a fermentation product, which is uniformly mixed with an adjuvant and water to obtain a plant growth regulator. The plant growth regulator prepared by the present invention can promote seed rooting, promote plant germination and seedling growth, facilitate uniform and strong seedlings, enhance plant drought and waterlogging resistance and stress resistance, improve disease, insect and bacteria resistance, significantly increase crop yield, and has safe and non-toxic ingredients, is easily degraded, does not cause environmental pollution, and has good application prospects.

Description

Technical Field

[0001] The present invention relates to the technical field of plant growth regulators, and in particular to a plant growth regulator and a preparation method thereof. Background Art

[0002] Phytohormones, also known as natural or endogenous plant hormones, are organic substances produced by plants themselves that act directly or indirectly on target organs or tissues to regulate plant growth. The physiological effects of phytohormones are complex, affecting root division, growth, and differentiation, germination, flowering, fruiting, and sexual differentiation. The amount of naturally synthesized phytohormones is very low, making it expensive to extract them in large quantities to increase crop yields. Consequently, through extensive research, scientists have discovered that chemical synthesis can produce organic compounds with similar structures and effects to natural phytohormones, known as plant growth regulators (PGRs). Once absorbed, PGRs stimulate the combined action of various active enzymes within the plant, thereby influencing the plant's physical and chemical processes.

[0003] Since the study of auxin in the 1930s, plant growth regulators have developed rapidly. Subsequently, gibberellins (GA), abscisic acid (ABA), cytokinins (CTK), and ethylene were discovered. Today, plant growth regulators are used in various fields, including cash crops, landscape gardening, and medicinal plants.

[0004] Plant growth regulators is a general term, which can be generally divided into three categories according to their effects on plant growth, namely plant growth promoters including auxins, cytokinins, gibberellins and brassinosteroids, plant growth inhibitors represented by cinnamic acid, coumarin, abscisic acid and salicylic acid, and plant growth retardants including chlormequat chloride, paclobutrazol (PP333) and uniclobutrazol.

[0005] With the world's arable land decreasing while the population is growing, coupled with the increasing frequency of natural disasters, plant growth regulators are gaining increasing attention in ensuring food security and increasing yields. However, traditional plant growth regulators, with their high residual content, high pollution levels, poor safety, and low utilization rates, do not meet the needs of ecological agriculture. Summary of the Invention

[0006] The purpose of the present invention is to provide a plant growth regulator and a preparation method thereof, which can promote seed rooting, promote plant germination and seedling growth, facilitate uniform and strong seedlings, enhance plant drought and waterlogging resistance and stress resistance, improve disease, insect and fungus resistance, significantly increase crop yield, and have safe and non-toxic ingredients, are easily degradable, will not cause environmental pollution, and have good application prospects.

[0007] The technical solution of the present invention is achieved as follows:

[0008] The invention provides a method for preparing a plant growth regulator. The method comprises the following steps: corn stalks, buckwheat seeds and rapeseed pollen are mixed, and yeast, Bacillus subtilis and Monascus purpureus are inoculated into the obtained enzymatic hydrolysis product through composite enzyme hydrolysis for fermentation to obtain a fermentation product, which is then uniformly mixed with an auxiliary agent and water to obtain the plant growth regulator. The composite enzyme is a composite mixture of cellulase, papain and pectinase.

[0009] As a further improvement of the present invention, the following steps are included:

[0010] S1. Preparation of the composition: Corn straw, buckwheat seeds and rapeseed pollen were washed, dried, crushed, and mixed to obtain a composition;

[0011] S2 enzymatic hydrolysis: The composition in step S1 is added to water, a complex enzyme is added for enzymatic hydrolysis, the enzyme is inactivated to obtain an enzymatic hydrolysis product; the complex enzyme is a complex mixture of cellulase, papain and pectinase;

[0012] S3. Activation of yeast and Bacillus subtilis: Yeast and Bacillus subtilis were inoculated into Gaughan's medium, activated, and then cultured into activated bacterial liquid;

[0013] S4. Activation of Monascus purpureus: Monascus purpureus was inoculated into PDA medium (potato dextrose agar medium) and activated to obtain activated Monascus purpureus;

[0014] S5. Preparation of a spore suspension of Monascus purpureus: Add sterile water to the activated Monascus purpureus obtained in step S4, scrape the surface of the agar with an inoculating loop to remove the spores, collect the spore suspension, shake, filter with sterile filter paper, count, and re-add sterile water to obtain a spore suspension of Monascus purpureus;

[0015] S6. Preparation of liquid culture medium: Dissolve the carbon source, nitrogen source, vitamins, and inorganic salts in sterile water, mix well, adjust the pH of the culture medium with PBS solution, and sterilize with ultraviolet light.

[0016] S7. Preparation of seed solution: The activated yeast culture solution and the activated Bacillus subtilis culture solution obtained in step S3, and the Monascus purpureus spore suspension obtained in step S5 were inoculated into the liquid culture medium obtained in step S6 and cultured to obtain a seed solution;

[0017] S8. Preparation of composite fermentation broth: The yeast obtained in step S7, Bacillus subtilis and Monascus purpurogenum seed solution were mixed in equal volumes and diluted to obtain a composite fermentation broth;

[0018] S9 fermentation: the composite fermentation broth obtained in step S8 was added to the enzymatic hydrolyzate obtained in step S2, fermented, filtered, the solid was washed with sterile water, the washing solution and the filtrate were combined, and freeze-dried to obtain a fermentation product;

[0019] S10. Preparation of plant growth regulator: The fermentation product obtained in step S9, the auxiliary agent and water are mixed evenly to obtain a plant growth regulator.

[0020] As a further improvement of the present invention, the mass ratio of corn straw, buckwheat seeds and rape pollen in step S1 is (5-10): (3-5): (2-7); the mass ratio of cellulase, papain and pectinase in the complex enzyme in step S2 is (5-12): (2-5): (1-3); the mass ratio of the composition, water and complex enzyme is 20: (70-100): (1-3); and the enzymatic hydrolysis conditions are 35-45°C for 3-5h.

[0021] As a further improvement of the present invention, the inoculation amount of the yeast and Bacillus subtilis in step S3 is 1-3% and 2-4% respectively; the activation culture condition is 35-40°C, and the culture time is 24-36h; the inoculation amount of Monascus purpurogenum in step S4 is 3-5%, the activation culture condition is 35-40°C, and the culture time is 24-36h; the spore concentration of the Monascus purpurogenum spore suspension in step S5 is 10 5 -10 6 pieces / mL.

[0022] As a further improvement of the present invention, the carbon source in step S6 is selected from one or more of molasses, glucose, maltose, lactose, sucrose, fructose, and soluble starch; the nitrogen source is selected from peptone, fish meal, ammonia water, urea, ammonium salt, nitrate, and amino acid; the vitamin is selected from one or more of vitamin C, vitamin B1, vitamin B2, vitamin B6, vitamin A, vitamin K, vitamin B12, vitamin D, and vitamin E; the inorganic salt is selected from sodium chloride, potassium chloride, calcium chloride, magnesium sulfate, ferric chloride, A mixture of one or more of zinc sulfate, copper sulfate, manganese sulfate, zinc chloride, copper chloride, and manganese chloride; the amino acid is selected from one or more of serine, glycine, threonine, valine, tryptophan, leucine, alanine, cysteine, methionine, lysine, isoleucine, and phenylalanine; the mass ratio of the carbon source, nitrogen source, vitamins, inorganic salts, and sterile water is (7-12):(2-5):(0.02-0.15):(0.4-1):100; and the pH value of the culture medium is adjusted to 6.7-7.0.

[0023] As a further improvement of the present invention, the culture conditions in step S7 are 35-40°C, the culture time is 36-48h, and the bacterial content of the bacterial seed liquid is 10 7 -10 8 cfu / mL; the dilution factor in step S8 is 100-200 times.

[0024] As a further improvement of the present invention, the mass ratio of the composite fermentation broth to the enzymatic hydrolysis product in step S9 is 2:(3-5); and the fermentation conditions are fermentation culture at 36-38° C. for 48-72 hours.

[0025] As a further improvement of the present invention, the auxiliary agent in step S10 includes at least one of a film-forming agent, a thickener, a dispersant, a wetting agent, and a surfactant; the film-forming agent is selected from at least one of seaweed powder, sodium hydroxymethyl cellulose, and chitin; the thickener is selected from at least one of xanthan gum, hydroxymethyl cellulose, gelatin, seaweed, gypsum, hydroxyethyl cellulose, methyl cellulose, magnesium aluminum silicate, and polyvinyl alcohol; the dispersant is selected from at least one of Morwet D-425, Terpense-2500, naphthalene sulfonate, lignin sulfonate, and polycarboxylate. at least one of; the wetting agent is selected from at least one of sodium lauryl sulfate, MorwetEFW, polyoxyethylene alkylphenol ether, polyoxyethylene fatty alcohol ether, and polyoxyethylene polyoxypropylene block copolymer; the surfactant is selected from at least one of sodium dodecylbenzenesulfonate, sodium dodecylsulfonate, sodium dodecyl sulfate, sodium hexadecylbenzenesulfonate, sodium hexadecylsulfonate, sodium hexadecylsulfonate, sodium octadecylbenzenesulfonate, sodium octadecylsulfonate, and Tween-80; the mass ratio of the fermentation product, the auxiliary agent, and the water is (2-5): (2-4): 100.

[0026] As a further improvement of the present invention, the present invention specifically comprises the following steps:

[0027] S1. Preparation of the composition: 5-10 parts by weight of corn straw, 3-5 parts by weight of buckwheat seeds and 2-7 parts by weight of rapeseed pollen were washed, dried, crushed, and mixed to obtain a composition;

[0028] S2. Enzymatic hydrolysis: 20 parts by weight of the composition in step S1 were added to 70-100 parts by weight of water, 1-3 parts by weight of the complex enzyme was added, and the enzymatic hydrolysis was carried out at 35-45 ° C for 3-5h, and the enzyme was inactivated at 105-110 ° C for 10-15min to obtain an enzymatic product; the complex enzyme was a complex mixture of cellulase, papain and pectinase in a mass ratio of (5-12): (2-5): (1-3);

[0029] S3. Activation of yeast and Bacillus subtilis: Yeast and Bacillus subtilis were inoculated into Gaughan's medium at inoculum levels of 1-3% and 2-4%, respectively, and incubated at 35-40°C for 24-36 hours to produce an activated culture solution.

[0030] S4. Activation of Monascus purpureus: Monascus purpureus was inoculated into PDA medium at an inoculum size of 3-5%, and activated and cultured at 35-40°C for 24-36h to obtain activated Monascus purpureus;

[0031] S5. Preparation of Monascus purpureus spore suspension: Add sterile water to the activated Monascus purpureus in step S4, scrape the agar surface with an inoculating loop to scrape off the spores, collect the spore suspension, shake for 10-20 minutes, filter with sterile filter paper, count, and re-add sterile water to obtain a Monascus purpureus spore suspension with a spore concentration of 10 5 -10 6 / mL;

[0032] S6. Preparation of liquid culture medium: 7-12 parts by weight of a carbon source, 2-5 parts by weight of a nitrogen source, 0.02-0.15 parts by weight of vitamins and 0.4-1 parts by weight of an inorganic salt were dissolved in 100 parts by weight of sterile water, mixed well, and the pH of the culture medium was adjusted to 6.7-7.0 with PBS solution and sterilized by ultraviolet light.

[0033] S7. Preparation of seed solution: The activated yeast solution and Bacillus subtilis solution obtained in step S3, and the Monascus purpureus spore suspension obtained in step S5 were inoculated into the liquid medium obtained in step S6, and cultured at 35-40 ° C for 36-48h to obtain a seed solution containing 10 7 -10 8 cfu / mL;

[0034] S8 composite fermentation broth preparation: the yeast obtained in step S7, Bacillus subtilis and Monascus purpurogenum seed solution was mixed in equal volumes and diluted 100-200 times to obtain a composite fermentation broth;

[0035] S9 fermentation: 2 parts by weight of the composite fermentation broth prepared in step S8 was added to 3-5 parts by weight of the enzymatic hydrolyzate prepared in step S2, and fermented at 36-38 ° C for 48-72h, filtered, and the solid was washed with sterile water, the washing solution and the filtrate were combined, and freeze-dried to obtain a fermentation product;

[0036] S10. Preparation of a plant growth regulator: 2-5 parts by weight of the fermentation product obtained in step S9, 2-4 parts by weight of an adjuvant and 100 parts by weight of water are mixed to obtain a plant growth regulator.

[0037] The present invention further protects a plant growth regulator prepared by the above preparation method.

[0038] The present invention has the following beneficial effects: corn stalks are rich in nutrients required for plant growth, such as nitrogen and carbon; buckwheat seeds are rich in plant growth promoters, including cytokinins, auxins, gibberellins, etc.; rapeseed pollen contains a variety of brassinolides; and the three are mixed and then enzymatically hydrolyzed, which has a synergistic effect. Under the action of a complex enzyme, including cellulase, papain, and pectinase, the cell walls of the corn stalks, buckwheat seeds, and rapeseed pollen are enzymatically hydrolyzed, and a large amount of nutrients are released. At the same time, protein substances are enzymatically hydrolyzed into small molecular short peptides and amino acids, thereby obtaining an enzymatic hydrolysis product containing rich ingredients.

[0039] Furthermore, yeast, Bacillus subtilis and Monascus purpurogenum are activated and cultured into a strain seed liquid, and the enzymatic hydrolysis product obtained above is fermented. Not only can the nutrients be further fermented to produce small molecules of carbon, nitrogen, potassium, phosphorus and other nutrients required for plant growth, but also substances that promote plant growth are produced during the fermentation process, including D-chiro-inositol (a type of vitamin B), vitamin B1, vitamin B12 and other growth-regulating vitamin substances, as well as brassinosteroids, auxin and other small molecules that promote cell division and plant growth. Thus, the obtained fermentation product can play a good role in promoting plant growth; in addition, more proline is also produced, which can significantly improve the stress resistance of the plant, and a large amount of phenolic substances such as chlorogenic acid, protocatechuic acid, catechins, etc. are produced, thereby improving the bacteria resistance and insect resistance, improving the disease resistance of the plant, significantly increasing crop yield, and improving the drought and flood resistance of the plant to a certain extent. Moreover, the plant growth regulator component is safe and non-toxic, easily degraded, and will not cause environmental pollution.

[0040] The plant growth regulator prepared by the present invention can promote seed rooting, promote plant germination and seedling growth, be beneficial to uniform and strong seedlings, enhance plant drought and waterlogging resistance and stress resistance, improve disease, insect and fungus resistance, significantly increase crop yield, and has safe and non-toxic ingredients, is easily degradable, does not cause environmental pollution, and has good application prospects. BRIEF DESCRIPTION OF THE DRAWINGS

[0041] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0042] Figure 1 This is a comparison chart of the proline content in the roots of each group in the test example of the present invention;

[0043] Figure 2 This is a comparison chart of the proline content in leaves of each group in the test example of the present invention. DETAILED DESCRIPTION

[0044] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0045] Corn straw was harvested corn straw of the corn variety "Weike 702"; buckwheat seeds were purchased from Jiangsu Changjing Seed Co., Ltd.; rapeseed pollen, 1 kg per bag, with a content greater than 99%, was purchased from Wuhan Penglei Biotechnology Co., Ltd.

[0046] Cellulase, brand Xiasheng FDY-2236, enzyme activity 5000U / g, papain, brand Xiasheng FDG-2203, enzyme activity 100000U / g, pectinase, brand Xiasheng FDY-2212, enzyme activity 60000U / g, were purchased from Xiasheng (Beijing) Biotechnology Development Co., Ltd.

[0047] Yeast was Angel high-activity dry yeast, purchased from Angel Yeast Co., Ltd.; Bacillus subtilis, 100 billion CFU / g, was purchased from Okobike Biotechnology Co., Ltd.; Monascus purpurogenum was ACCC30352 Monascus purpurogenum, purchased from Shanghai Fuxiang Biotechnology Co., Ltd.

[0048] Example 1

[0049] This embodiment provides a method for preparing a plant growth regulator, which specifically includes the following steps:

[0050] S1. Preparation of the composition: 150 g corn straw, 90 g buckwheat seeds and 60 g rapeseed pollen were washed, dried at 70 ° C for 2 h, crushed, and mixed to obtain a composition;

[0051] S2. Enzymatic hydrolysis: 300 g of the composition in step S1 was added to 1400 g of water, 20 g of complex enzyme was added, and the enzymatic hydrolysis was carried out at 35 ° C for 3 h, and the enzyme was inactivated at 105 ° C for 10 min to obtain an enzymatic product; the complex enzyme was a complex mixture of cellulase, papain and pectinase in a mass ratio of 5:2:1;

[0052] S3. Activation of yeast and Bacillus subtilis: Yeast and Bacillus subtilis were inoculated into Gaughan's medium at inoculum levels of 1% and 2%, respectively, and incubated at 35°C for 24 h to produce activated culture fluids.

[0053] S4. Activation of Monascus purpureus: Monascus purpureus was inoculated into PDA medium at an inoculum size of 3% and cultured at 35°C for 24 hours to obtain activated Monascus purpureus;

[0054] S5. Preparation of Monascus purpureus spore suspension: Add sterile water to the activated Monascus purpureus in step S4, scrape the agar surface with an inoculating loop to scrape off the spores, collect the spore suspension, shake for 10 minutes, filter with sterile filter paper, count, and re-add sterile water to obtain a Monascus purpureus spore suspension with a spore concentration of 10 5 / mL;

[0055] S6. Preparation of liquid culture medium: 7g glucose, 2g peptone, 0.01g vitamin C, 0.01g vitamin B1 and 0.2g sodium chloride, 0.1g zinc sulfate, 0.1g copper sulfate were dissolved in 100g sterile water, mixed well, and the pH value of the culture medium was adjusted to 6.7 with PBS solution and sterilized by ultraviolet light.

[0056] S7. Preparation of seed solution: The activated yeast and Bacillus subtilis spores obtained in step S3 and the Monascus purpureus spore suspension obtained in step S5 were inoculated into the liquid culture medium obtained in step S6 and cultured at 35 ° C for 36 h to obtain a seed solution of bacteria, wherein the bacterial content of each seed solution was 10 7 cfu / mL;

[0057] S8 preparation of composite fermentation broth: the yeast obtained in step S7, Bacillus subtilis and Monascus purpurogenum seed solution were mixed in equal volumes and diluted 100 times to obtain a composite fermentation broth;

[0058] S9 fermentation: 200g of the composite fermentation broth obtained in step S8 was added to 300g of the enzymatic hydrolyzate obtained in step S2, and fermented at 36 ° C for 48h, filtered, and the solid was washed with sterile water, the washing solution and the filtrate were combined, and freeze-dried to obtain a fermentation product;

[0059] S10. Preparation of a plant growth regulator: 200 g of the fermentation product obtained in step S9, 200 g of an auxiliary agent, and 10 kg of water were mixed to obtain a plant growth regulator. The auxiliary agent included seaweed powder, sodium lauryl sulfate, and Morwet D-425 in a mass ratio of 3:1:1.

[0060] Example 2

[0061] This embodiment provides a method for preparing a plant growth regulator, which specifically includes the following steps:

[0062] S1. Preparation of the composition: 200 g corn straw, 100 g buckwheat seeds and 140 g rapeseed pollen were washed, dried at 70 ° C for 2 h, crushed, and mixed to obtain a composition;

[0063] S2. Enzymatic hydrolysis: 440 g of the composition in step S1 was added to 2200 g of water, 66 g of complex enzyme was added, and the enzymatic hydrolysis was carried out at 45 ° C for 5 h, and the enzyme was inactivated at 110 ° C for 15 min to obtain an enzymatic product; the complex enzyme was a complex mixture of cellulase, papain and pectinase in a mass ratio of 12:5:3;

[0064] S3. Activation of yeast and Bacillus subtilis: Yeast and Bacillus subtilis were inoculated into Gaughan's medium at inoculum levels of 3% and 4%, respectively, and incubated at 40°C for 36 h to produce activated culture fluids.

[0065] S4. Activation of Monascus purpureus: Monascus purpureus was inoculated into PDA medium at an inoculum size of 5% and activated at 40°C for 36 h to obtain activated Monascus purpureus;

[0066] S5. Preparation of Monascus purpureus spore suspension: Add sterile water to the activated Monascus purpureus in step S4, scrape the agar surface with an inoculating loop to scrape off the spores, collect the spore suspension, shake for 20 minutes, filter with sterile filter paper, count, and re-add sterile water to obtain a Monascus purpureus spore suspension with a spore concentration of 10 6 / mL;

[0067] S6. Preparation of liquid culture medium: 6g glucose, 6g maltose, 3g peptone, 2g fish meal, 0.05g vitamin B1, 0.05g vitamin B2, 0.05g vitamin B6, 0.4g sodium chloride, 0.2g magnesium chlorosulfate, 0.2g ferric chloride, 0.2g zinc sulfate were dissolved in 100g sterile water, mixed well, and the pH value of the culture medium was adjusted to 7.0 with PBS solution and sterilized by ultraviolet light.

[0068] S7. Preparation of seed solution: The activated yeast and Bacillus subtilis spores obtained in step S3 and the Monascus purpureus spore suspension obtained in step S5 were inoculated into the liquid culture medium obtained in step S6 and cultured at 40 ° C for 48 h to obtain a seed solution of bacteria, wherein the bacterial content of each seed solution was 10 8 cfu / mL;

[0069] S8 composite fermentation broth preparation: the yeast obtained in step S7, Bacillus subtilis and Monascus purpurogenum seed solution was mixed in equal volumes and diluted 200 times to obtain a composite fermentation broth;

[0070] S9 fermentation: 200g of the composite fermentation broth obtained in step S8 was added to 500g of the enzymatic hydrolyzate obtained in step S2, and the fermentation was cultured at 38 ° C for 72h, filtered, and the solid was washed with sterile water, the washing solution and the filtrate were combined, and freeze-dried to obtain a fermentation product;

[0071] S10. Preparation of a plant growth regulator: 500 g of the fermentation product obtained in step S9, 400 g of an auxiliary agent, and 10 kg of water were mixed to obtain a plant growth regulator. The auxiliary agent included seaweed powder, gelatin, and sodium hexadecylbenzenesulfonate in a mass ratio of 3:2:1.

[0072] Example 3

[0073] This embodiment provides a method for preparing a plant growth regulator, which specifically includes the following steps:

[0074] S1. Preparation of the composition: 70 g corn straw, 40 g buckwheat seeds and 50 g rapeseed pollen were washed, dried at 70 ° C for 2 h, crushed, and mixed to obtain a composition;

[0075] S2. Enzymatic hydrolysis: 160 g of the composition in step S1 was added to 640 g of water, 16 g of complex enzyme was added, and the enzymatic hydrolysis was carried out at 40 ° C for 4 h, and the enzyme was inactivated at 107 ° C for 12 min to obtain an enzymatic product; the complex enzyme was a complex mixture of cellulase, papain and pectinase in a mass ratio of 7:3.5:2;

[0076] S3. Activation of yeast and Bacillus subtilis: Yeast and Bacillus subtilis were inoculated into Gaughan's medium at 2% and 3% inoculum, respectively, and incubated at 37°C for 30 h to produce activated culture fluids.

[0077] S4. Activation of Monascus purpureus: Monascus purpureus was inoculated into PDA medium at an inoculum size of 4% and activated at 37°C for 30 h to obtain activated Monascus purpureus;

[0078] S5. Preparation of Monascus purpureus spore suspension: Add sterile water to the activated Monascus purpureus in step S4, scrape the agar surface with an inoculating loop to scrape off the spores, collect the spore suspension, shake for 15 minutes, filter with sterile filter paper, count, and re-add sterile water to obtain a Monascus purpureus spore suspension with a spore concentration of 10 6 / mL;

[0079] S6. Preparation of liquid culture medium: Dissolve 6 g glucose, 2 g sucrose, 2 g fructose, 2 g urea, 1.5 g ammonium nitrate, 0.05 g vitamin B1, 0.05 g vitamin K, 0.3 g potassium chloride, 0.1 g copper sulfate, 0.1 g manganese sulfate, and 0.1 g zinc chloride in 100 g sterile water, mix well, adjust the medium pH to 6.85 with PBS solution, and sterilize with ultraviolet light.

[0080] S7. Preparation of seed solution: The activated yeast and Bacillus subtilis spores obtained in step S3 and the Monascus purpureus spore suspension obtained in step S5 were inoculated into the liquid culture medium obtained in step S6 and cultured at 37 ° C for 42 h to obtain a seed solution containing 10 8 cfu / mL;

[0081] S8 composite fermentation broth preparation: the yeast obtained in step S7, Bacillus subtilis and Monascus purpurogenum seed solution of equal volume mixed and diluted 150 times to obtain a composite fermentation broth;

[0082] S9 fermentation: 180g of the composite fermentation broth obtained in step S8 was added to 360g of the enzymatic hydrolyzate obtained in step S2, and the fermentation was cultured at 37 ° C for 56h, filtered, and the solid was washed with sterile water, the washing solution and the filtrate were combined, and freeze-dried to obtain a fermentation product;

[0083] S10. Preparation of a plant growth regulator: 350 g of the fermentation product obtained in step S9, 300 g of an auxiliary agent, and 10 kg of water were mixed to obtain a plant growth regulator. The auxiliary agent included sodium carboxymethylcellulose, xanthan gum, Morwet D-425, and Tween-80 in a mass ratio of 5:2:2:1.

[0084] Example 4

[0085] Compared with Example 3, the composite enzyme is a compound mixture of cellulase and pectinase, with a mass ratio of 10.5:2, and other conditions remain unchanged.

[0086] Example 5

[0087] Compared with Example 3, the composite enzyme is a compound mixture of papain and pectinase, with a mass ratio of 10.5:2, and other conditions remain unchanged.

[0088] Comparative Example 1

[0089] Compared with Example 3, buckwheat seeds were not added in step S1, and other conditions remained unchanged.

[0090] The specific steps include:

[0091] S1. Preparation of the composition: 70 g corn straw and 90 g rapeseed pollen were washed, dried at 70 ° C for 2 h, crushed, and mixed to obtain a composition;

[0092] S2. Enzymatic hydrolysis: 160 g of the composition in step S1 was added to 640 g of water, 16 g of complex enzyme was added, and the enzymatic hydrolysis was carried out at 40 ° C for 4 h, and the enzyme was inactivated at 107 ° C for 12 min to obtain an enzymatic product; the complex enzyme was a complex mixture of cellulase, papain and pectinase in a mass ratio of 7:3.5:2;

[0093] S3. Activation of yeast and Bacillus subtilis: Yeast and Bacillus subtilis were inoculated into Gaughan's medium at 2% and 3% inoculum, respectively, and incubated at 37°C for 30 h to produce activated culture fluids.

[0094] S4. Activation of Monascus purpureus: Monascus purpureus was inoculated into PDA medium at an inoculum size of 4% and activated at 37°C for 30 h to obtain activated Monascus purpureus;

[0095] S5. Preparation of Monascus purpureus spore suspension: Add sterile water to the activated Monascus purpureus in step S4, scrape the agar surface with an inoculating loop to scrape off the spores, collect the spore suspension, shake for 15 minutes, filter with sterile filter paper, count, and re-add sterile water to obtain a Monascus purpureus spore suspension with a spore concentration of 10 6 / mL;

[0096] S6. Preparation of liquid culture medium: Dissolve 6 g glucose, 2 g sucrose, 2 g fructose, 2 g urea, 1.5 g ammonium nitrate, 0.05 g vitamin B1, 0.05 g vitamin K, 0.3 g potassium chloride, 0.1 g copper sulfate, 0.1 g manganese sulfate, and 0.1 g zinc chloride in 100 g sterile water, mix well, adjust the medium pH to 6.85 with PBS solution, and sterilize with ultraviolet light.

[0097] S7. Preparation of seed solution: The activated yeast and Bacillus subtilis spores obtained in step S3 and the Monascus purpureus spore suspension obtained in step S5 were inoculated into the liquid culture medium obtained in step S6 and cultured at 37 ° C for 42 h to obtain a seed solution containing 10 8 cfu / mL;

[0098] S8 composite fermentation broth preparation: the yeast obtained in step S7, Bacillus subtilis and Monascus purpurogenum seed solution of equal volume mixed and diluted 150 times to obtain a composite fermentation broth;

[0099] S9 fermentation: 180g of the composite fermentation broth obtained in step S8 was added to 360g of the enzymatic hydrolyzate obtained in step S2, and the fermentation was cultured at 37 ° C for 56h, filtered, and the solid was washed with sterile water, the washing solution and the filtrate were combined, and freeze-dried to obtain a fermentation product;

[0100] S10. Preparation of a plant growth regulator: 35 g of the fermentation product obtained in step S9, 30 g of an adjuvant, and 1000 g of water were mixed to obtain a plant growth regulator. The adjuvant included sodium carboxymethylcellulose, xanthan gum, Morwet D-425, and Tween-80 in a mass ratio of 5:2:2:1.

[0101] Comparative Example 2

[0102] Compared with Example 3, rape pollen was not added in step S1, and other conditions remained unchanged.

[0103] The specific steps include:

[0104] S1. Preparation of the composition: 70 g corn straw and 90 g buckwheat seeds were washed, dried at 70 ° C for 2 h, crushed, and mixed to obtain a composition;

[0105] S2. Enzymatic hydrolysis: 160 g of the composition of step S1 was added to 640 g of water, 16 g of the complex enzyme was added, and the mixture was hydrolyzed at 40°C for 4 h, and the enzyme was inactivated at 107°C for 12 min to obtain an enzymatic hydrolysis product; the complex enzyme was a compound mixture of cellulase, papain, and pectinase in a mass ratio of 7:3.5:2; S3. Activation of yeast and Bacillus subtilis: Yeast and Bacillus subtilis were inoculated into Gould's medium at inoculation rates of 2% and 3%, respectively, and activated and cultured at 37°C for 30 h, and then cultured into an activated bacterial culture liquid;

[0106] S4. Activation of Monascus purpureus: Monascus purpureus was inoculated into PDA medium at an inoculum size of 4% and activated at 37°C for 30 h to obtain activated Monascus purpureus;

[0107] S5. Preparation of Monascus purpureus spore suspension: Add sterile water to the activated Monascus purpureus in step S4, scrape the agar surface with an inoculating loop to scrape off the spores, collect the spore suspension, shake for 15 minutes, filter with sterile filter paper, count, and re-add sterile water to obtain a Monascus purpureus spore suspension with a spore concentration of 10 6 / mL;

[0108] S6. Preparation of liquid culture medium: Dissolve 6 g glucose, 2 g sucrose, 2 g fructose, 2 g urea, 1.5 g ammonium nitrate, 0.05 g vitamin B1, 0.05 g vitamin K, 0.3 g potassium chloride, 0.1 g copper sulfate, 0.1 g manganese sulfate, and 0.1 g zinc chloride in 100 g sterile water, mix well, adjust the medium pH to 6.85 with PBS solution, and sterilize with ultraviolet light.

[0109] S7. Preparation of seed solution: The activated yeast and Bacillus subtilis spores obtained in step S3 and the Monascus purpureus spore suspension obtained in step S5 were inoculated into the liquid culture medium obtained in step S6 and cultured at 37 ° C for 42 h to obtain a seed solution containing 10 8 cfu / mL;

[0110] S8 composite fermentation broth preparation: the yeast obtained in step S7, Bacillus subtilis and Monascus purpurogenum seed solution of equal volume mixed and diluted 150 times to obtain a composite fermentation broth;

[0111] S9 fermentation: 180g of the composite fermentation broth obtained in step S8 was added to 360g of the enzymatic hydrolyzate obtained in step S2, and the fermentation was cultured at 37 ° C for 56h, filtered, and the solid was washed with sterile water, the washing solution and the filtrate were combined, and freeze-dried to obtain a fermentation product;

[0112] S10. Preparation of a plant growth regulator: 35 g of the fermentation product obtained in step S9, 30 g of an adjuvant, and 1000 g of water were mixed to obtain a plant growth regulator. The adjuvant included sodium carboxymethylcellulose, xanthan gum, Morwet D-425, and Tween-80 in a mass ratio of 5:2:2:1.

[0113] Comparative Example 3

[0114] Compared with Example 3, step S2 enzymatic hydrolysis was not performed, and other conditions remained unchanged.

[0115] The specific steps include:

[0116] S1. Preparation of the composition: 70 g corn straw, 40 g buckwheat seeds and 50 g rapeseed pollen were washed, dried at 70 ° C for 2 h, crushed, and mixed to obtain a composition;

[0117] S2. Activation of Yeast and Bacillus subtilis: Yeast and Bacillus subtilis were inoculated into Gaughan's medium at 2% and 3% inoculum, respectively, and incubated at 37°C for 30 h to produce activated culture fluids.

[0118] S3. Activation of Monascus purpureus: Monascus purpureus was inoculated into PDA medium at a 4% inoculation rate and cultured at 37°C for 30 h to obtain activated Monascus purpureus;

[0119] S4. Preparation of Monascus purpureus spore suspension: Add sterile water to the activated Monascus purpureus in step S3, scrape the agar surface with an inoculating loop to scrape off the spores, collect the spore suspension, shake for 15 minutes, filter with sterile filter paper, count, and re-add sterile water to obtain a Monascus purpureus spore suspension with a spore concentration of 10 6 / mL;

[0120] S5. Preparation of liquid culture medium: Dissolve 6 g glucose, 2 g sucrose, 2 g fructose, 2 g urea, 1.5 g ammonium nitrate, 0.05 g vitamin B1, 0.05 g vitamin K, 0.3 g potassium chloride, 0.1 g copper sulfate, 0.1 g manganese sulfate, and 0.1 g zinc chloride in 100 g sterile water, mix well, adjust the medium pH to 6.85 with PBS solution, and sterilize with ultraviolet light.

[0121] S6. Preparation of seed solution: The activated yeast and Bacillus subtilis spores obtained in step S2 and the Monascus purpureus spore suspension obtained in step S4 were inoculated into the liquid culture medium obtained in step S5 and cultured at 37 ° C for 42 h to obtain a seed solution containing 10 8 cfu / mL;

[0122] S7. Preparation of composite fermentation broth: The yeast obtained in step S6, Bacillus subtilis and Monascus purpurogenum seed solution were mixed in equal volumes and diluted 150 times to obtain a composite fermentation broth;

[0123] S8 fermentation: 200g of the composite fermentation broth prepared in step S7 was added to 400g of the composition prepared in step S1, and fermented at 37 ° C for 56h, filtered, and the solid was washed with sterile water, and the washing solution and the filtrate were combined and freeze-dried to obtain a fermentation product;

[0124] S9. Preparation of a plant growth regulator: 35 g of the fermentation product obtained in step S9, 30 g of an adjuvant, and 1000 g of water were mixed to obtain a plant growth regulator. The adjuvant included sodium carboxymethylcellulose, xanthan gum, Morwet D-425, and Tween-80 in a mass ratio of 5:2:2:1.

[0125] Comparative Example 4

[0126] Compared with Example 3, Bacillus subtilis was not added and other conditions remained unchanged.

[0127] The specific steps include:

[0128] S1. Preparation of the composition: 70 g corn straw, 40 g buckwheat seeds and 50 g rapeseed pollen were washed, dried at 70 ° C for 2 h, crushed, and mixed to obtain a composition;

[0129] S2. Enzymatic hydrolysis: 160 g of the composition of step S1 was added to 640 g of water, 16 g of the complex enzyme was added, and the mixture was hydrolyzed at 40°C for 4 h, and the enzyme was inactivated at 107°C for 12 min to obtain an enzymatic hydrolysis product; the complex enzyme was a compound mixture of cellulase, papain, and pectinase in a mass ratio of 7:3.5:2; S3. Yeast activation: The yeast was inoculated into Gould's medium at an inoculum size of 5%, activated and cultured at 37°C for 30 h, and then cultivated into an activated bacterial culture liquid;

[0130] S4. Activation of Monascus purpureus: Monascus purpureus was inoculated into PDA medium at an inoculum size of 4% and activated at 37°C for 30 h to obtain activated Monascus purpureus;

[0131] S5. Preparation of Monascus purpureus spore suspension: Add sterile water to the activated Monascus purpureus in step S4, scrape the agar surface with an inoculating loop to scrape off the spores, collect the spore suspension, shake for 15 minutes, filter with sterile filter paper, count, and re-add sterile water to obtain a Monascus purpureus spore suspension with a spore concentration of 10 6 / mL;

[0132] S6. Preparation of liquid culture medium: Dissolve 6 g glucose, 2 g sucrose, 2 g fructose, 2 g urea, 1.5 g ammonium nitrate, 0.05 g vitamin B1, 0.05 g vitamin K, 0.3 g potassium chloride, 0.1 g copper sulfate, 0.1 g manganese sulfate, and 0.1 g zinc chloride in 100 g sterile water, mix well, adjust the medium pH to 6.85 with PBS solution, and sterilize with ultraviolet light.

[0133] S7. Preparation of seed solution: The yeast activated seed solution obtained in step S3 and the Monascus purpureus spore suspension obtained in step S5 were inoculated into the liquid culture medium prepared in step S6 and cultured at 37 ° C for 42 h to obtain a seed solution of bacteria, wherein the bacterial content of each seed solution was 10 8 cfu / mL;

[0134] S8. Preparation of composite fermentation broth: The yeast and Monascus purpurogenum seed solution obtained in step S7 were mixed in equal volumes and diluted 150 times to obtain a composite fermentation broth;

[0135] S9 fermentation: 180g of the composite fermentation broth obtained in step S8 was added to 360g of the enzymatic hydrolyzate obtained in step S2, and the fermentation was cultured at 37 ° C for 56h, filtered, and the solid was washed with sterile water, the washing solution and the filtrate were combined, and freeze-dried to obtain a fermentation product;

[0136] S10. Preparation of a plant growth regulator: 35 g of the fermentation product obtained in step S9, 30 g of an adjuvant, and 1000 g of water were mixed to obtain a plant growth regulator. The adjuvant included sodium carboxymethylcellulose, xanthan gum, Morwet D-425, and Tween-80 in a mass ratio of 5:2:2:1.

[0137] Comparative Example 5

[0138] Compared with Example 3, no yeast was added and other conditions remained unchanged.

[0139] The specific steps include:

[0140] S1. Preparation of the composition: 70 g corn straw, 40 g buckwheat seeds and 50 g rapeseed pollen were washed, dried at 70 ° C for 2 h, crushed, and mixed to obtain a composition;

[0141] S2. Enzymatic hydrolysis: 160 g of the composition in step S1 was added to 640 g of water, 16 g of the complex enzyme was added, and the mixture was hydrolyzed at 40°C for 4 h and inactivated at 107°C for 12 min to obtain an enzymatic hydrolysis product; the complex enzyme was a mixture of cellulase, papain, and pectinase in a mass ratio of 7:3.5:2; S3. Activation of Bacillus subtilis: Bacillus subtilis was inoculated into Gould's medium at an inoculum size of 5%, activated and cultured at 37°C for 30 h, and then cultured into an activated bacterial culture liquid;

[0142] S4. Activation of Monascus purpureus: Monascus purpureus was inoculated into PDA medium at an inoculum size of 4% and activated at 37°C for 30 h to obtain activated Monascus purpureus;

[0143] S5. Preparation of Monascus purpureus spore suspension: Add sterile water to the activated Monascus purpureus in step S4, scrape the agar surface with an inoculating loop to scrape off the spores, collect the spore suspension, shake for 15 minutes, filter with sterile filter paper, count, and re-add sterile water to obtain a Monascus purpureus spore suspension with a spore concentration of 10 6 / mL;

[0144] S6. Preparation of liquid culture medium: Dissolve 6 g glucose, 2 g sucrose, 2 g fructose, 2 g urea, 1.5 g ammonium nitrate, 0.05 g vitamin B1, 0.05 g vitamin K, 0.3 g potassium chloride, 0.1 g copper sulfate, 0.1 g manganese sulfate, and 0.1 g zinc chloride in 100 g sterile water, mix well, adjust the medium pH to 6.85 with PBS solution, and sterilize with ultraviolet light.

[0145] S7. Preparation of seed solution: The activated Bacillus subtilis solution obtained in step S3 and the Monascus purpurogenum spore suspension obtained in step S5 were inoculated into the liquid culture medium prepared in step S6 and cultured at 37 ° C for 42 h to obtain a seed solution of bacteria, wherein the bacterial content of each seed solution was 10 8 cfu / mL;

[0146] S8. Preparation of composite fermentation broth: The Bacillus subtilis and Monascus purpurogenum seed solution obtained in step S7 were mixed in equal volumes and diluted 150 times to obtain a composite fermentation broth;

[0147] S9 fermentation: 180g of the composite fermentation broth obtained in step S8 was added to 360g of the enzymatic hydrolyzate obtained in step S2, and the fermentation was cultured at 37 ° C for 56h, filtered, and the solid was washed with sterile water, the washing solution and the filtrate were combined, and freeze-dried to obtain a fermentation product;

[0148] S10. Preparation of a plant growth regulator: 35 g of the fermentation product obtained in step S9, 30 g of an adjuvant, and 1000 g of water were mixed to obtain a plant growth regulator. The adjuvant included sodium carboxymethylcellulose, xanthan gum, Morwet D-425, and Tween-80 in a mass ratio of 5:2:2:1.

[0149] Comparative Example 6

[0150] Compared with Example 3, no Monascus purpurogenum was added, and other conditions remained unchanged.

[0151] The specific steps include:

[0152] S1. Preparation of the composition: 70 g corn straw, 40 g buckwheat seeds and 50 g rapeseed pollen were washed, dried at 70 ° C for 2 h, crushed, and mixed to obtain a composition;

[0153] S2. Enzymatic hydrolysis: 160 g of the composition of step S1 was added to 640 g of water, 16 g of the complex enzyme was added, and the mixture was hydrolyzed at 40°C for 4 h, and the enzyme was inactivated at 107°C for 12 min to obtain an enzymatic hydrolysis product; the complex enzyme was a compound mixture of cellulase, papain, and pectinase in a mass ratio of 7:3.5:2; S3. Activation of yeast and Bacillus subtilis: Yeast and Bacillus subtilis were inoculated into Gould's medium at inoculation rates of 2% and 3%, respectively, and activated and cultured at 37°C for 30 h, and then cultured into an activated bacterial culture liquid;

[0154] S4. Preparation of liquid culture medium: Dissolve 6 g glucose, 2 g sucrose, 2 g fructose, 2 g urea, 1.5 g ammonium nitrate, 0.05 g vitamin B1, 0.05 g vitamin K, 0.3 g potassium chloride, 0.1 g copper sulfate, 0.1 g manganese sulfate, and 0.1 g zinc chloride in 100 g sterile water, mix well, adjust the pH of the culture medium to 6.85 with PBS solution, and sterilize with ultraviolet light.

[0155] S5. Preparation of seed solution: The activated yeast solution and the activated Bacillus subtilis solution obtained in step S3 were inoculated into the liquid culture medium prepared in step S4 and cultured at 37°C for 42h to obtain a seed solution containing 10 8 cfu / mL;

[0156] S6 composite fermentation broth preparation: the yeast obtained in step S5, Bacillus subtilis seed solution of equal volume mixed and diluted 150 times to obtain a composite fermentation broth;

[0157] S7 fermentation: 200g of the composite fermentation broth obtained in step S6 was added to 400g of the enzymatic hydrolysate obtained in step S2, and fermented at 37 ° C for 56h, filtered, and the solid was washed with sterile water, the washing solution and the filtrate were combined, and freeze-dried to obtain a fermentation product;

[0158] S8. Preparation of a plant growth regulator: 35 g of the fermentation product obtained in step S7, 30 g of an adjuvant, and 1000 g of water were mixed to obtain a plant growth regulator. The adjuvant included sodium carboxymethylcellulose, xanthan gum, Morwet D-425, and Tween-80 in a mass ratio of 5:2:2:1.

[0159] Comparative Example 7

[0160] Compared with Example 3, the fermentation step was not performed and other conditions remained unchanged.

[0161] The specific steps include:

[0162] S1. Preparation of the composition: 70 g corn straw, 40 g buckwheat seeds and 50 g rapeseed pollen were washed, dried at 70 ° C for 2 h, crushed, and mixed to obtain a composition;

[0163] S2. Enzymatic hydrolysis: Add 160 g of the composition from step S1 to 640 g of water, add 16 g of a complex enzyme, and perform enzymatic hydrolysis at 40°C for 4 hours. Inactivate the enzyme at 107°C for 12 minutes to obtain an enzymatic hydrolysis product. The complex enzyme is a mixture of cellulase, papain, and pectinase in a mass ratio of 7:3.5:2. S3. Preparation of a plant growth regulator: Mix 35 g of the enzymatic hydrolysis product from step S2, 30 g of an adjuvant, and 1000 g of water to obtain a plant growth regulator. The adjuvant includes sodium carboxymethylcellulose, xanthan gum, Morwet D-425, and Tween-80 in a mass ratio of 5:2:2:1.

[0164] Test Example 1

[0165] 1. Test on promoting germination of corn seeds

[0166] Corn seeds with full grains and uniform size were selected and grouped into 14 groups, each with 100 grains. The variety was "Weike 702", which were respectively Example 1-5 groups, Comparative Example 1-7 groups, water groups, and indoleacetic acid groups. Each group of corn seeds was placed in a culture dish covered with double-layer filter paper, and 25 mL of sample liquid was injected for the first time. The seeds were cultured in a constant temperature box at 25±2°C in the dark, and 2 mL of sample liquid was newly injected every day.

[0167] Sample liquid: Example 1-5 group and Comparative Example 1-7 group were the plant growth regulators prepared in Example 1-5 and Comparative Example 1-7, the indoleacetic acid group was injected with 50 μg / mL indoleacetic acid aqueous solution, and the clean water group was clean water.

[0168] Observe the germination of the embryo after 5 days, and take the embryo length exceeding 1 / 2 of the seed length as the germination standard; calculate the germination rate.

[0169] Germination rate (%) = number of germinated seeds / total number of test samples × 100%;

[0170] The results are shown in Table 1.

[0171] Table 1

[0172] Group Germination rate (%) Example 1 100 Example 2 100 Example 3 100 Example 4 84 Example 5 80 Comparative Example 1 67 Comparative Example 2 64 Comparative Example 3 75 Comparative Example 4 77 Comparative Example 5 82 Comparative Example 6 70 Comparative Example 7 62 clear water 45 Indoleacetic acid 71

[0173] As can be seen from the above table, the plant growth regulators prepared in Examples 1-3 of the present invention have a good effect of promoting corn seed germination, which is better than indoleacetic acid.

[0174] 2. Growth promotion test

[0175] After the corn seeds germinated, five of the best-growing seeds from each group were selected and potted. Watered daily with the sample solution at the appropriate concentration to ensure normal growth. At the five-leaf stage, plant height, taproot length, and leaf fresh weight were measured, and the growth promotion rate was calculated. Free proline content in roots and leaves was also measured.

[0176] Promotion rate of plant height, taproot length and leaf fresh weight (%) = (plant height, taproot length and leaf fresh weight of this group - plant height, taproot length and leaf fresh weight of clear water group) / plant height, taproot length and leaf fresh weight of clear water group × 100%.

[0177] The results are shown in Table 2 and Figure 1-2 .

[0178] Table 2

[0179]

[0180] As can be seen from the above table, the plant growth regulators prepared in Examples 1-3 of the present invention have a good effect of promoting the growth of corn seedlings, which is better than indoleacetic acid.

[0181] Proline (Pro) is a component of plant protein and is widely present in a free state throughout plants. Under stress conditions such as drought and salinity, many plants accumulate large amounts of proline. Accumulated proline not only acts as an osmotic regulator in plant cytoplasm, but also plays an important role in stabilizing the structure of biomolecules, reducing cellular acidity, detoxifying ammonia, and acting as an energy reservoir to regulate cellular redox potential. Under adverse conditions (drought, salinity, heat, cold, and freezing), proline content in plants increases significantly. Proline content in plants reflects, to a certain extent, a plant's stress tolerance, with drought-resistant varieties often accumulating higher amounts of proline. Therefore, measuring proline content can be used as a physiological indicator for drought-resistance breeding. Furthermore, due to its strong hydrophilicity, proline stabilizes protoplasmic colloids and metabolic processes within tissues, thereby lowering the freezing point and preventing cellular dehydration. Under low-temperature conditions, an increase in proline in plant tissues can improve cold tolerance and, therefore, can be used as a physiological indicator for cold-resistance breeding.

[0182] Depend on Figure 1-2 It can be seen that the plant growth regulators prepared in Examples 1-3 of the present invention can significantly increase the proline content of plants, thereby promoting plant germination and seedling growth, making the seedlings uniform and strong, enhancing the drought and waterlogging resistance and stress resistance of plants, improving the disease, insect and fungus resistance, and significantly increasing crop yields.

[0183] Compared with Example 3, in Comparative Examples 1 and 2, no buckwheat seeds or rapeseed pollen were added in Step S1, resulting in significantly reduced rooting, sprouting, and growth-promoting effects. Corn stalks are rich in nutrients such as nitrogen and carbon, which are essential for plant growth. Buckwheat seeds are rich in plant growth regulators, including cytokinins, auxins, and gibberellins. Rapeseed pollen contains a variety of brassinolides. Mixing these three and then enzymatically hydrolyzing them produces a large amount of growth-promoting substances, such as brassinolides, cytokinins, auxins, and gibberellins, resulting in a synergistic effect.

[0184] Compared with Example 3, the composite enzyme is a composite mixture of cellulase and pectinase or a composite mixture of papain and pectinase, and its rooting, sprouting and growth effects are reduced, and the proline content in the plant body is reduced; Compared with Example 3, step S2 enzymolysis is not performed in Comparative Example 3, and its rooting, sprouting and growth effects are significantly reduced, and the proline content in the plant body is significantly reduced. The present invention performs enzymolysis after mixing corn stalks, buckwheat seeds and rapeseed pollen, which has a synergistic effect. Under the action of the composite enzyme, including cellulase, papain and pectinase, the cell wall is enzymolyzed, and a large amount of nutrients are released. At the same time, protein substances are enzymolyzed into small molecule short peptides, amino acids, including proline, etc., so as to obtain an enzymolysis product containing rich ingredients, which can significantly promote plant growth and improve its stress resistance.

[0185] Compared with Example 3, Comparative Examples 4 and 5 did not add Bacillus subtilis or yeast, and Comparative Example 6 did not add Monascus purpurogenum compared with Example 3. The rooting, sprouting, and growth promoting effects of the comparative examples were reduced, and the proline content in the plants was reduced. Compared with Example 3, Comparative Example 7 did not perform the fermentation step, and the rooting, sprouting, and growth promoting effects of the comparative examples were significantly reduced, and the proline content in the plants was significantly reduced. Yeast, Bacillus subtilis and Monascus purpurogenum are activated and then cultured into a strain liquid. The enzymatic hydrolysis product obtained above is fermented. Not only can the nutrients be further fermented to produce small molecules of carbon, nitrogen, potassium, phosphorus and other nutrients required for plant growth, but also substances that promote plant growth are produced during the fermentation process, including D-chiro-inositol (a type of vitamin B), vitamin B1, vitamin B12 and other growth-regulating vitamins, as well as brassinosteroids, auxin and other small molecules that promote cell division and plant growth. Therefore, the obtained fermentation product can effectively promote plant growth. In addition, more proline is produced, which can significantly improve the stress resistance of plants, and a large amount of phenolic substances such as chlorogenic acid, protocatechuic acid, catechins, etc. are produced, thereby improving the bacteria resistance and insect resistance, improving the disease resistance of plants, significantly increasing crop yields, and improving the drought and flood resistance of plants to a certain extent. The plant growth regulator component is safe and non-toxic, easily degradable, and will not cause environmental pollution.

[0186] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A method for preparing a plant growth regulator, characterized in that: Corn stalks, buckwheat seeds and rapeseed pollen are mixed to obtain a composition, and a composite fermentation liquid obtained by yeast, Bacillus subtilis and Monascus purpureus is inoculated into the hydrolysis product obtained by composite enzyme hydrolysis, and fermentation is carried out to obtain a fermentation product, which is uniformly mixed with an auxiliary agent and water to obtain a plant growth regulator; the composite enzyme is a composite mixture of cellulase, papain and pectinase; the mass ratio of the corn stalks, buckwheat seeds and rapeseed pollen is (5-10): (3-5): (2-7); the mass ratio of the cellulase, papain and pectinase in the composite enzyme is (5-10): (3-5): (2-7); The mass ratio of the composition, water and complex enzyme is 20: (70-100): (1-3); the enzymatic hydrolysis condition is 35-45° C. for 3-5 hours; the mass ratio of the complex fermentation broth and the enzymatic hydrolysis product is 2: (3-5); the fermentation condition is 36-38° C. for 48-72 hours; the yeast is Angel high-activity dry yeast, purchased from Angel Yeast Co., Ltd.; the Bacillus subtilis is purchased from Okobaike Biotechnology Co., Ltd.; the Monascus purpureus is ACCC 30352 Monascus purpureus, purchased from Shanghai Fuxiang Biotechnology Co., Ltd.; the mass ratio of the fermentation product, auxiliary agent and water is (2-5): (2-4):

100.

2. The preparation method according to claim 1, characterized in that The following steps are involved: S1. Preparation of the composition: Corn straw, buckwheat seeds and rapeseed pollen were washed, dried, crushed, and mixed to obtain a composition; S2 enzymatic hydrolysis: The composition in step S1 is added to water, a complex enzyme is added for enzymatic hydrolysis, the enzyme is inactivated to obtain an enzymatic hydrolysis product; the complex enzyme is a complex mixture of cellulase, papain and pectinase; S3. Activation of yeast and Bacillus subtilis: Yeast and Bacillus subtilis were inoculated into Gaughan's medium, activated, and then cultured into activated bacterial liquid; S4. Activation of Monascus purpureus: The Monascus purpureus was inoculated into PDA medium and activated to obtain activated Monascus purpureus; S5. Preparation of a spore suspension of Monascus purpureus: Add sterile water to the activated Monascus purpureus obtained in step S4, scrape the surface of the agar with an inoculating loop to remove the spores, collect the spore suspension, shake, filter with sterile filter paper, count, and re-add sterile water to obtain a spore suspension of Monascus purpureus; S6. Preparation of liquid culture medium: Dissolve the carbon source, nitrogen source, vitamins, and inorganic salts in sterile water, mix well, adjust the pH of the culture medium with PBS solution, and sterilize with ultraviolet light. S7. Preparation of seed solution: The activated yeast culture solution and the activated Bacillus subtilis culture solution obtained in step S3, and the Monascus purpureus spore suspension obtained in step S5 were inoculated into the liquid culture medium obtained in step S6 and cultured to obtain a seed solution; S8. Preparation of composite fermentation broth: The yeast obtained in step S7, Bacillus subtilis and Monascus purpurogenum seed solution were mixed in equal volumes and diluted to obtain a composite fermentation broth; S9 fermentation: the composite fermentation broth obtained in step S8 was added to the enzymatic hydrolyzate obtained in step S2, fermented, filtered, the solid was washed with sterile water, the washing solution and the filtrate were combined, and freeze-dried to obtain a fermentation product; S10. Preparation of plant growth regulator: The fermentation product obtained in step S9, the auxiliary agent and water are mixed evenly to obtain a plant growth regulator.

3. The preparation method according to claim 2, characterized in that The inoculation amount of yeast and Bacillus subtilis in step S3 is 1-3% and 2-4% respectively; the activation culture condition is 35-40°C and the culture time is 24-36h; the inoculation amount of Monascus purpureus in step S4 is 3-5%, the activation culture condition is 35-40°C and the culture time is 24-36h; the spore concentration of the Monascus purpureus spore suspension in step S5 is 10 5 -10 6 pieces / mL.

4. The preparation method according to claim 2, characterized in that In step S6, the carbon source is selected from one or more of molasses, glucose, maltose, lactose, sucrose, fructose, and soluble starch; the nitrogen source is selected from peptone, fish meal, ammonia water, urea, ammonium salt, nitrate, and amino acid; the vitamin is selected from one or more of vitamin C, vitamin B1, vitamin B2, vitamin B6, vitamin A, vitamin K, vitamin B12, vitamin D, and vitamin E; the inorganic salt is selected from sodium chloride, potassium chloride, calcium chloride, magnesium sulfate, ferric chloride, zinc sulfate, sulfuric acid A mixture of one or more of copper, manganese sulfate, zinc chloride, copper chloride, and manganese chloride; the amino acid is selected from one or more of serine, glycine, threonine, valine, tryptophan, leucine, alanine, cysteine, methionine, lysine, isoleucine, and phenylalanine; the mass ratio of the carbon source, nitrogen source, vitamins, inorganic salts, and sterile water is (7-12): (2-5): (0.02-0.15): (0.4-1): 100; the pH value of the culture medium is adjusted to 6.7-7.

0.

5. The preparation method according to claim 2, characterized in that The culture conditions in step S7 are 35-40°C, the culture time is 36-48h, and the bacterial content of the bacterial seed liquid is 10 7 -10 8 cfu / mL; the dilution factor in step S8 is 100-200 times.

6. The preparation method according to claim 2, characterized in that The auxiliary agent in step S10 includes at least one of a film-forming agent, a thickener, a dispersant, a wetting agent, and a surfactant; the film-forming agent is selected from at least one of seaweed powder, sodium hydroxymethyl cellulose, and chitin; the thickener is selected from at least one of xanthan gum, hydroxymethyl cellulose, gelatin, seaweed, gypsum, hydroxyethyl cellulose, methyl cellulose, magnesium aluminum silicate, and polyvinyl alcohol; the dispersant is selected from at least one of Morwet D-425, Terspense-2500, naphthalene sulfonate, lignin sulfonate, and polycarboxylate; the wetting agent is selected from at least one of sodium lauryl sulfate, Morwet EFW, polyoxyethylene alkylphenol ether, polyoxyethylene fatty alcohol ether, and polyoxyethylene polyoxypropylene block copolymer; the surfactant is selected from at least one of sodium dodecylbenzenesulfonate, sodium dodecylsulfonate, sodium dodecyl sulfate, sodium hexadecylbenzenesulfonate, sodium hexadecylsulfonate, sodium hexadecylsulfonate, sodium hexadecylsulfonate, sodium octadecylbenzenesulfonate, sodium octadecylsulfonate, and Tween-80.

7. The preparation method according to claim 2, characterized in that The specific steps include: S1. Preparation of the composition: 5-10 parts by weight of corn straw, 3-5 parts by weight of buckwheat seeds and 2-7 parts by weight of rapeseed pollen were washed, dried, crushed, and mixed to obtain a composition; S2. Enzymatic hydrolysis: 20 parts by weight of the composition in step S1 were added to 70-100 parts by weight of water, 1-3 parts by weight of a complex enzyme was added, and the enzymatic hydrolysis was carried out at 35-45°C for 3-5 hours, and the enzyme was inactivated at 105-110°C for 10-15 minutes to obtain an enzymatic hydrolysis product; the complex enzyme was a mixture of cellulase, papain and pectinase in a mass ratio of (5-12): (2-5): (1-3); S3. Activation of Yeast and Bacillus subtilis: Inoculate 1-3% and 2-4% yeast into Gaughan's medium, respectively, and incubate at 35-40°C for 24-36 hours to produce an activated culture. S4. Activation of Monascus purpureus: Inoculate Monascus purpureus into PDA medium at an inoculum size of 3-5% and activate and culture at 35-40°C for 24-36 hours to obtain activated Monascus purpureus; S5. Preparation of Monascus purpureus spore suspension: Add sterile water to the activated Monascus purpureus in step S4, scrape the agar surface with an inoculating loop to scrape off the spores, collect the spore suspension, shake for 10-20 minutes, filter with sterile filter paper, count, and re-add sterile water to obtain a Monascus purpureus spore suspension with a spore concentration of 10 5 -10 6 / mL; S6. Preparation of liquid culture medium: 7-12 parts by weight of a carbon source, 2-5 parts by weight of a nitrogen source, 0.02-0.15 parts by weight of vitamins and 0.4-1 parts by weight of an inorganic salt were dissolved in 100 parts by weight of sterile water, mixed well, and the pH of the culture medium was adjusted to 6.7-7.0 with PBS solution and sterilized by ultraviolet light. S7. Preparation of seed solution: The activated yeast solution and Bacillus subtilis solution obtained in step S3, and the Monascus purpureus spore suspension obtained in step S5 were inoculated into the liquid medium obtained in step S6, and cultured at 35-40 ° C for 36-48h to obtain a seed solution containing 10 7 -10 8 cfu / mL; S8 composite fermentation broth preparation: the yeast obtained in step S7, Bacillus subtilis and Monascus purpurogenum seed solution was mixed in equal volumes and diluted 100-200 times to obtain a composite fermentation broth; S9 fermentation: 2 parts by weight of the composite fermentation broth prepared in step S8 was added to 3-5 parts by weight of the enzymatic hydrolyzate prepared in step S2, and fermented at 36-38 ° C for 48-72h, filtered, and the solid was washed with sterile water, the washing solution and the filtrate were combined, and freeze-dried to obtain a fermentation product; S10. Preparation of a plant growth regulator: 2-5 parts by weight of the fermentation product obtained in step S9, 2-4 parts by weight of an adjuvant and 100 parts by weight of water are mixed to obtain a plant growth regulator.

8. A plant growth regulator prepared by the preparation method according to any one of claims 1 to 7.

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