Plant growth regulator for improving stress tolerance of crops

Abstract

The application discloses a kind of plant growth regulators for improving the stress resistance of crops, belongs to plant growth regulator technical field.It is made of the following weight parts of raw materials: modified black fungus polysaccharide 10-20 parts, grape wine processing solid by-product fermentation liquor 30-50 parts, sodium complex nitrophenol 0.05-0.15 parts, trisodium citrate 1-3 parts, gamma-aminobutyric acid 1-3 parts, accelerator 0.3-0.6 parts;The accelerator is nanochitosan and sodium cocoyl glutamate according to 1:5 mass ratio composition.The plant growth regulator prepared in the application can significantly improve the stress resistance of crops after use, improve the ability of crops to resist saline-alkali, drought and low temperature stress.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of plant growth regulators, and particularly relates to a plant growth regulator for improving the stress resistance of crops. BACKGROUND

[0002] Plants often suffer from various stresses during growth, including biological stress (pests, weeds, etc.) and abiotic stress (drought, high and low temperature, high salt and alkali, weak light, heavy metals, etc.), which causes damage to plants and even death in severe cases. The damage caused by stress to plants mainly includes membrane system damage, active oxygen damage, destruction of protective enzyme systems, dehydration of plant cells, decreased photosynthesis enzyme activity, unstable respiration, and enhanced catabolism, etc. Plants have also formed adaptability and resistance to adverse environments during growth and evolution, i.e. the stress resistance of plants.

[0003] Salt-alkali stress generally includes salt stress and alkali stress. Salt stress mainly refers to stress caused by neutral salt mainly in the form of NaCl and Na2SO4 to plants, while alkali stress refers to stress caused by alkaline salt mainly in the form of Na2CO3 and NaHCO3 to plants. Under salt-alkali stress, the root cells of crops are harmed by osmotic stress and high pH, resulting in the lack of mineral elements such as phosphorus and magnesium in the body of crops. The specific harm to crops is manifested in the inhibition of normal growth of crops, the influence on the absorption of water and nutrients, the direct damage to plant cell structure, and the destruction of soil structure, etc., which seriously affects the yield and quality of crops.

[0004] The damage caused by low temperature stress includes damage caused by low temperature above 0℃ (i.e. cold damage) and damage caused by low temperature below 0℃ (i.e. freezing damage). Cold damage slows down the growth of plants, destroys the water balance in the body, causes wilting due to water loss, increases the permeability of cell membranes, causes organic matter to exude, and causes metabolic disorders in plants. The synthesis of chlorophyll is inhibited, the leaves turn yellow and lose green, photosynthesis enzyme activity is inhibited, and the photosynthesis rate decreases. Protein synthesis is reduced, and the content of endogenous hormones is increased. Freezing damage includes extracellular freezing and intracellular freezing, which mainly causes damage to protoplasm through ice crystals, causes damage to the cell membrane system, loss of membrane selectivity, exudation of organic matter, changes in enzyme activity, and destruction of protein structure. Low temperature stress seriously affects the growth and development of crops.

[0005] Stress environment seriously affects the yield and quality of crops, and the method of improving the stress resistance of crops by externally applying plant growth regulators is widely used in agricultural production. Plant growth regulators can regulate different stages of plant growth and development, such as promoting seed germination, rooting, flowering, fruit setting, ripening, dormancy, etc., and can also improve crop quality and enhance the ability of plants to resist cold, drought, salinity and disease and insect pests. Plant growth regulators are widely used in crop stress resistance cultivation due to their low cost, small amount and significant effect. Currently, the commonly used plant growth regulators are mainly chemical synthetic substances, such as auxin, gibberellin acid, abscisic acid, sodium complex nitrophenol, etc. However, a single plant growth regulator often has poor effect on regulating plant growth, especially on enhancing the stress resistance of crops, so there is an urgent need for a plant growth regulator that can improve the stress resistance of crops in order to achieve the best regulation effect. SUMMARY

[0006] The purpose of the present application is to provide a plant growth regulator for improving the stress resistance of crops. The modified Auricularia auricular polysaccharide extracted by enzyme method is reacted with sodium selenite to obtain selenium Auricularia auricular polysaccharide. The introduction of selenium element improves the biological activity of Auricularia auricular polysaccharide. After compounding with other components, the stress resistance of crops can be significantly improved, and the ability of crops to resist salinity, drought and low temperature stress can be improved.

[0007] To achieve the above technical purpose, the technical scheme adopted by the present application is:

[0008] A plant growth regulator for improving the stress resistance of crops, which is made of the following raw materials by weight: modified Auricularia auricular polysaccharide 5-15 parts, grape wine processing solid by-product fermentation liquid 20-30 parts, sodium complex nitrophenol 0.05-0.15 parts, trisodium citrate 1-3 parts, gamma-aminobutyric acid 1-3 parts, and promoter 0.3-0.6 parts. The promoter is composed of nano-chitosan and sodium cocoyl glutamate in a mass ratio of 1:5.

[0009] Preferably, the modified Auricularia auricular polysaccharide is prepared by the following method:

[0010] (1) After drying and crushing Auricularia auricular to pass through a 100-mesh sieve, deionized water and Auricularia auricular powder are added according to the solid-liquid ratio, homogenized, the pH is adjusted to 4.0-4.5, 5wt% complex enzyme solution is added, enzyme hydrolysis is carried out for 2h, then the temperature is raised to 55℃ and the pH is adjusted to 6.0-6.5, 2% saccharifying enzyme is added, enzyme hydrolysis is carried out for 1h, then the pH is adjusted to 6.5-7.0, mannase and neutral protease are added, and enzyme hydrolysis is carried out at 40-45℃ for 3-4h, high-temperature enzyme inactivation is carried out, and enzyme hydrolysis liquid is obtained; the enzyme hydrolysis liquid is centrifuged, the supernatant is taken, 3 times the amount of 95% ethanol is added, and high-speed centrifugal separation is carried out after standing for 24h, the obtained precipitate is dried to obtain Auricularia auricular polysaccharide;

[0011] (2) Take 5g black fungus polysaccharide and dissolve it in 200mL 2mol / L sodium hydroxide solution, stir until dissolved, then add 1.5wt% chloroacetic acid, heat to 50℃ and stir for 4h, then cool to room temperature to obtain a mixture, freeze-dry the mixture to obtain acidified black fungus polysaccharide;

[0012] (3) Mix the acidified black fungus polysaccharide with sodium selenite in a certain proportion to obtain a mixture, add 1% nitric acid solution to the mixture until dissolved, then add BaCl2, stir uniformly, and react at 70℃ water bath for 4-5h, then add appropriate amount of saturated sodium sulfate solution to remove barium chloride, centrifugal separation, adjust the pH value to neutral, add 3 times amount of 95% ethanol, stand for 24h, then high-speed centrifugal separation, dry the precipitate to obtain modified black fungus polysaccharide.

[0013] Preferably, the ratio of black fungus powder to deionized water in step (1) is 1g:30ml; the cellulase in the complex enzyme solution contains 10-15g / L, and the pectinase contains 15-20g / L; the addition amount of mannanase is 0.5-1g / L, and the addition amount of neutral protease is 1.5-2g / L.

[0014] Preferably, the enzyme activity of cellulase is 5000U / g, the enzyme activity of pectinase is 40000U / g; the enzyme activity of saccharifying enzyme is 10000U / g, the enzyme activity of mannanase is 50000U / g, and the enzyme activity of neutral protease is 60000U / g.

[0015] Preferably, the mass ratio of acidified black fungus polysaccharide to sodium selenite in step (3) is 3:1, and the ratio of the amount of 1% nitric acid solution, the mixture, BaCl2, and saturated sodium sulfate solution is 50ml:1g:0.2g:3-5ml.

[0016] Preferably, the wine processing solid by-product fermentation liquid is prepared by the following method:

[0017] a. Dry the wine processing solid by-product, crush it through an 80 mesh sieve to obtain a solid powder, add a complex hydrolytic enzyme solution to the solid powder, and hydrolyze at 35-40℃ for 6-8h, then inactivate at high temperature to obtain a hydrolysis liquid;

[0018] b. Add Bacillus licheniformis and Lactobacillus plantarum to the hydrolysis liquid, adjust the temperature to 30-35℃, ferment for 24h, then filter, and take the filtrate.

[0019] Preferably, the ratio of the amount of solid powder to complex hydrolytic enzyme solution in step a is 1g:1.5-2ml.

[0020] Preferably, the complex hydrolytic enzyme solution contains cellulase 10-15 g / L, bromelain 20-25 g / L; the cellulase enzyme activity is 5000 U / g, and the bromelain enzyme activity is 100,000 U / g.

[0021] Preferably, the amount of bacillus licheniformis added in step b is 10-15 g / L, and the amount of lactobacillus plantarum added is 20-30 g / L.

[0022] Preferably, the viable count of bacillus licheniformis is 100 billion / g, and the viable count of lactobacillus plantarum is 10 billion / g.

[0023] The raw materials used in the application are all commercially available products, unless otherwise specified, wherein the nanochitosan is purchased from Xi'an Qiyue Biological Technology Co., Ltd., with a particle size of 100-150 nm; the bacillus licheniformis is purchased from Guangxi Nongbao Biological Engineering Co., Ltd.; and the lactobacillus plantarum is purchased from Shandong Pinggao Pharmaceutical Co., Ltd.

[0024] The plant growth regulator for improving the stress resistance of crops obtained in the application is a liquid agent, which can be mixed with fertilizer or used alone, when mixed, the dosage is 2-3% of the mass of the fertilizer; when used alone, it can be sprayed or root applied to crops after dilution, and the dosage is 10 kg / acre.

[0025] The solid by-products of grape wine processing mainly include discarded grape skins, seeds, pectin and tartar, and also include dead yeast cells, fruit peel fragments, fruit pulp particles and other solid substances. Traditionally, these solid waste residues are discharged to a landfill or used as organic fertilizer. However, these methods not only cause environmental burden, but also may spread pathogenic bacteria and chemical residues. In the application, the solid waste residues are subjected to enzymatic hydrolysis and fermentation treatment. Firstly, the enzymatic hydrolysis process decomposes complex organic matter into small molecular substances such as simple sugars and amino acids. On the one hand, these substances are more easily absorbed and utilized by microorganisms for fermentation, so as to produce more abundant and more biologically active metabolites, which have a significant effect on enhancing the stress resistance of crops. On the other hand, these substances can be directly absorbed and utilized by crops, promoting the growth and development of crops and improving the stress resistance of crops.

[0026] The modified auricularia auricular polysaccharide used in the application is seleniumized auricularia auricular polysaccharide, and the introduction of selenium element improves the biological activity of auricularia auricular polysaccharide and the promoting effect on the growth of plants is more significant. After the modified auricularia auricular polysaccharide is combined with the fermentation liquor of the solid by-products of grape wine processing, the two components have a synergistic effect, and the ability of crops to resist saline-alkali, drought and low temperature stress is significantly improved.

[0027] The promoter of the application contains nanochitosan and disodium cocoyl glutamate in a specific ratio, which plays a role in promoting the penetration and absorption of various active substances, wherein the nanochitosan can also be used as a carrier to load a small amount of incomplete anthocyanin in the fermentation liquor of the solid by-product of grape wine processing onto the surface of plants and be better absorbed and utilized by plants through the penetration-promoting performance of disodium cocoyl glutamate, so as to optimize the nutrient distribution and physiological metabolism in the plant body, thereby improving the stress resistance and yield of the plant.

[0028] The application has the advantages that: the application realizes the recycling of resources by utilizing the solid by-product of grape wine processing for fermentation, which not only can reduce the emission of waste, reduce environmental pollution and improve the economic benefits of enterprises; the modified auricularia auricular polysaccharide is combined with the fermentation liquor of the solid by-product of grape wine processing, and the two synergistically improve the ability of crops to resist saline-alkali, drought and low-temperature stress. The plant growth regulator prepared by the application has the advantages of being safer and having fewer side effects, and when used as a plant growth regulator or foliar fertilizer for crops, it can effectively activate the defense mechanism of the plant body, relieve the damage of crops caused by abiotic stress, improve the stress resistance and improve the quality of crops. BRIEF DESCRIPTION OF DRAWINGS

[0029] Figure 1 For the SOD enzyme activity of the small rape seedlings in test example 3 of the application;

[0030] Figure 2 For the CAT enzyme activity of the small rape seedlings in test example 3 of the application;

[0031] Figure 3 For the POD enzyme activity of the small rape seedlings in test example 3 of the application. DETAILED DESCRIPTION

[0032] The technical solutions of the application will be further described below in combination with specific embodiments, but are not limited thereto.

[0033] Example 1

[0034] A plant growth regulator for improving the stress resistance of crops, which is prepared from the following raw materials in parts by weight: modified auricularia auricular polysaccharide 5 kg, fermentation liquor of solid by-product of grape wine processing 20 kg, sodium complex nitrophenol 0.05 kg, trisodium citrate 1 kg, gamma-aminobutyric acid 1 kg, and promoter 0.3 kg; the promoter is composed of nanochitosan and disodium cocoyl glutamate in a mass ratio of 1:5.

[0035] The modified auricularia auricular polysaccharide is prepared by the following method:

[0036] (1) Take dried black fungus, crush it through a 100-mesh sieve, and add deionized water and black fungus powder at a ratio of 30 ml: 1 g of material liquid. Homogenize the mixture, adjust the pH to 4.0, and then add a 5 wt% complex enzyme solution. Enzymatically hydrolyze for 2 h, then increase the temperature to 55°C and adjust the pH to 6.0. Add 2% saccharifying enzyme and enzymatically hydrolyze for 1 h. Then adjust the pH to 6.5, add mannanase and neutral protease, and enzymatically hydrolyze at 40°C for 3-4 h. Inactivate the enzymes at high temperature to obtain an enzymatic hydrolysate. Centrifuge the enzymatic hydrolysate, take the supernatant, add 3 times the amount of 95% ethanol, and let it stand for 24 h. Then centrifuge at high speed to obtain a precipitate. Dry the precipitate to obtain black fungus polysaccharide.

[0037] The complex enzyme solution contains 10 g / L of cellulase and 15 g / L of pectinase. The mannanase is added at a dosage of 0.5 g / L, and the neutral protease is added at a dosage of 1.5 g / L. The cellulase has an enzyme activity of 5000 U / g, and the pectinase has an enzyme activity of 40000 U / g. The saccharifying enzyme has an enzyme activity of 10000 U / g, the mannanase has an enzyme activity of 50000 U / g, and the neutral protease has an enzyme activity of 60000 U / g.

[0038] (2) Dissolve 5 g of black fungus polysaccharide in 200 mL of 2 mol / L sodium hydroxide solution, stir until dissolved, then add 1.5 wt% chloroacetic acid, heat to 50°C and stir for 4 h. Then cool to room temperature to obtain a mixed solution. Freeze-dry the mixed solution to obtain acidified black fungus polysaccharide.

[0039] (3) Mix the acidified black fungus polysaccharide with sodium selenite at a mass ratio of 3:1 to obtain a mixture. Dissolve the mixture in a 1% nitric acid solution, then add BaCl2, stir uniformly, and react in a 70°C water bath for 4-5 h. After the reaction is complete, add an appropriate amount of saturated sodium sulfate solution to remove barium chloride. Centrifuge to obtain a supernatant, adjust the pH to neutral, add 3 times the amount of 95% ethanol, let it stand for 24 h, and then centrifuge at high speed. Dry the obtained precipitate to obtain modified black fungus polysaccharide. The amount of the 1% nitric acid solution, the mixture, BaCl2, and the saturated sodium sulfate solution is 50 ml: 1 g: 0.2 g: 3 ml.

[0040] The grape wine processing solid by-product fermentation liquid is prepared by the following method:

[0041] a. Dry the grape wine processing solid by-product, crush it through an 80-mesh sieve to obtain a solid powder, add a complex hydrolytic enzyme solution to the solid powder, and enzymatically hydrolyze at 35°C for 8 h. Inactivate the enzymes at high temperature to obtain an enzymatic hydrolysate;

[0042] b. Add Bacillus licheniformis and Lactobacillus plantarum to the enzymatic hydrolysate, adjust the temperature to 30°C, and ferment for 24 h. Then filter the mixture to obtain a filtrate.

[0043] The ratio of the solid powder to the complex hydrolytic enzyme solution in step a is 1g:1.5ml.

[0044] The complex hydrolytic enzyme solution contains cellulase 10g / L and bromelain 20g / L; the cellulase enzyme activity is 5000U / g, and the bromelain enzyme activity is 100000U / g.

[0045] The amount of bacillus licheniformis added in step b is 10g / L, and the amount of lactobacillus plantarum added is 20g / L; the bacillus licheniformis viable count is 100 billion / g; and the lactobacillus plantarum viable count is 10 billion / g.

[0046] Example 2

[0047] A plant growth regulator for improving the stress resistance of crops, which is made of the following raw materials in parts by weight: modified black fungus polysaccharide 15kg, grape wine processing solid by-product fermentation liquor 30kg, sodium complex nitrophenol 0.15kg, trisodium citrate 3kg, gamma-aminobutyric acid 3kg, and promoter 0.6kg; the promoter is composed of nanochitosan and sodium cocoyl glutamate in a mass ratio of 1:5.

[0048] The modified black fungus polysaccharide is prepared by the following method:

[0049] (1) After drying, the black fungus is crushed to pass through a 100-mesh sieve, and deionized water and black fungus powder are added in a ratio of 30ml:1g of liquid to solid, and homogenized. The pH is adjusted to 4.5, and then 5wt% of a complex enzyme solution is added. After 2h of enzymatic hydrolysis, the temperature is increased to 55℃ and the pH is adjusted to 6.5. Then 2% of saccharifying enzyme is added, and the pH is adjusted to 7.0. Mannanase and neutral protease are added, and the enzymatic hydrolysis is carried out at 45℃ for 3-4h. The enzyme is inactivated at high temperature. The enzyme hydrolysis liquor is obtained. The enzyme hydrolysis liquor is centrifuged, and the supernatant is taken and added with 3 times the amount of 95% ethanol. After standing for 24h, high-speed centrifugal separation is carried out. The obtained precipitate is dried to obtain black fungus polysaccharide.

[0050] The complex enzyme solution contains cellulase 15g / L and pectinase 20g / L; the amount of mannase added is 1g / L, and the amount of neutral protease added is 2g / L; the enzyme activity of cellulase is 5000U / g, the enzyme activity of pectinase is 40000U / g; the enzyme activity of saccharifying enzyme is 10000U / g, the enzyme activity of mannase is 50000U / g, and the enzyme activity of neutral protease is 60000U / g.

[0051] (2) take 5g black fungus polysaccharide is dissolved in 200ml 2mol / l sodium hydroxide solution, stirring until dissolved completely, then add 1.5wt% chloroacetic acid, heated to 50℃ stirring reaction 4h, after cooling to room temperature to get mixed solution, the mixed solution freeze drying into powder to obtain acidified black fungus polysaccharide;

[0052] (3) the acidified black fungus polysaccharide and sodium selenite are mixed according to the mass ratio of 3:1 to obtain a mixture, the mixture is dissolved completely by adding 1% nitric acid solution, then BaCl2 is added, stirred uniformly, and reacted at 70℃ water bath for 4-5h, after the reaction is completed, a proper amount of saturated sodium sulfate solution is added to remove BaCl2, centrifugal separation is carried out to take the supernatant, the pH value is adjusted to neutral, 3 times of 95% ethanol is added, and high speed centrifugal separation is carried out after standing for 24h, and the obtained precipitate is dried to obtain modified black fungus polysaccharide; the amount ratio of the 1% nitric acid solution, the mixture, BaCl2 and the saturated sodium sulfate solution is 50ml:1g:0.2g:5ml.

[0053] The wine processing solid by-product fermentation liquid is prepared by the following method:

[0054] a. the wine processing solid by-product is dried, crushed and passed through an 80 mesh sieve to obtain a solid powder, a complex hydrolytic enzyme solution is added to the solid powder for enzymolysis at 40℃ for 6h, and high temperature inactivation is carried out to obtain an enzymolysis liquid;

[0055] b. Bacillus licheniformis and Lactobacillus plantarum are added to the enzymolysis liquid, the temperature is adjusted to 35℃, and fermentation is carried out for 24h, then filtration is carried out, and the filtrate is obtained.

[0056] The amount ratio of the solid powder to the complex hydrolytic enzyme solution in step a is 1g:2ml.

[0057] The complex hydrolytic enzyme solution contains cellulase 15g / L and bromelain 25g / L; the cellulase enzyme activity is 5000U / g, and the bromelain enzyme activity is 100000U / g.

[0058] The amount of Bacillus licheniformis added in step b is 15g / L, and the amount of Lactobacillus plantarum added is 30g / L; the viable count of the Bacillus licheniformis is 100 billion / g; and the viable count of the Lactobacillus plantarum is 10 billion / g.

[0059] Example 3

[0060] A plant growth regulator for improving the stress resistance of crops, which is prepared from the following raw materials in parts by weight: modified black fungus polysaccharide 10kg, wine processing solid by-product fermentation liquid 25kg, sodium nitrophenolate 0.1kg, trisodium citrate 2kg, gamma-aminobutyric acid 2kg, and promoter 0.5kg; the promoter is composed of nanochitosan and sodium cocoyl glutamate according to a mass ratio of 1:5.

[0061] The modified Auricularia auricular polysaccharide is prepared by the following method:

[0062] (1) After drying Auricularia auricular, it is crushed to pass through a 100-mesh sieve, and deionized water and Auricularia auricular powder are added at a ratio of 30 ml:1 g of liquid to solid, and homogenized. The pH is adjusted to 4.5, and then 5 wt% of a complex enzyme solution is added. After 2 h of enzymatic hydrolysis, the temperature is increased to 55°C and the pH is adjusted to 6.0. Then 2% of saccharifying enzyme is added, and the enzymatic hydrolysis is carried out for 1 h. Then the pH is adjusted to 7.0, and mannase and neutral protease are added. The enzymatic hydrolysis is carried out at 45°C for 3-4 h, and the enzyme is inactivated at high temperature. The enzyme solution is obtained. The enzyme solution is centrifuged, and the supernatant is taken. After adding 3 times the amount of 95% ethanol and standing for 24 h, high-speed centrifugal separation is carried out. The obtained precipitate is dried to obtain Auricularia auricular polysaccharide.

[0063] The complex enzyme solution contains 13 g / L of cellulase and 18 g / L of pectinase. The addition amount of mannase is 0.8 g / L, and the addition amount of neutral protease is 1.8 g / L. The enzyme activity of cellulase is 5000 U / g, and the enzyme activity of pectinase is 40000 U / g. The enzyme activity of saccharifying enzyme is 10000 U / g, the enzyme activity of mannase is 50000 U / g, and the enzyme activity of neutral protease is 60000 U / g.

[0064] (2) 5 g of Auricularia auricular polysaccharide is dissolved in 200 mL of 2 mol / L sodium hydroxide solution, stirred until completely dissolved, then 1.5 wt% chloroacetic acid is added, heated to 50°C and stirred for 4 h, then cooled to room temperature to obtain a mixed solution. The mixed solution is freeze-dried into powder to obtain acidified Auricularia auricular polysaccharide.

[0065] (3) The acidified Auricularia auricular polysaccharide is mixed with sodium selenite at a mass ratio of 3:1 to obtain a mixture. The mixture is dissolved completely by adding 1% nitric acid solution, and then BaCl2 is added. After stirring uniformly, the reaction is carried out at 70°C water bath for 4-5 h. After the reaction is completed, a proper amount of saturated sodium sulfate solution is added to remove barium chloride. After centrifugal separation, the supernatant is taken, the pH value is adjusted to neutral, 3 times the amount of 95% ethanol is added and stood for 24 h, and then high-speed centrifugal separation is carried out. The obtained precipitate is dried to obtain modified Auricularia auricular polysaccharide. The amount ratio of 1% nitric acid solution, mixture, BaCl2, and saturated sodium sulfate solution is 50 ml:1 g:0.2 g:4 ml.

[0066] The grape wine processing solid by-product fermentation liquid is prepared by the following method:

[0067] a. The grape wine processing solid by-product is dried and crushed to pass through an 80-mesh sieve to obtain a solid powder. A complex hydrolytic enzyme solution is added to the solid powder, and the enzymatic hydrolysis is carried out at 40°C for 6 h. After high-temperature inactivation, the enzyme solution is obtained.

[0068] b. After adding bacillus licheniformis and lactobacillus plantarum to the enzymatic hydrolysate, the temperature is adjusted to 35℃ and fermentation is carried out for 24 hours, then filtration is carried out, and the filtrate is obtained.

[0069] The ratio of the solid powder to the complex hydrolytic enzyme solution in step a is 1g:2ml.

[0070] The complex hydrolytic enzyme solution contains cellulase 12g / L and bromelain 23g / L; the cellulase enzyme activity is 5000U / g, and the bromelain enzyme activity is 100,000U / g.

[0071] The bacillus licheniformis is added in an amount of 12g / L, and the lactobacillus plantarum is added in an amount of 25g / L; the bacillus licheniformis has a viable count of 100 billion / g; and the lactobacillus plantarum has a viable count of 10 billion / g.

[0072] Example 4

[0073] A plant growth regulator for improving the stress resistance of crops, which is prepared from the following raw materials in parts by weight: modified black fungus polysaccharide 12kg, grape wine processing solid by-product fermentation liquor 25kg, sodium complex nitrophenol 0.08kg, trisodium citrate 1.5kg, gamma-aminobutyric acid 1.5kg, and a promoter 0.4kg; the promoter is composed of nanochitosan and sodium cocoyl glutamate in a mass ratio of 1:5.

[0074] The modified black fungus polysaccharide is prepared by the following method:

[0075] (1) After drying, the black fungus is crushed to pass through a 100-mesh sieve, deionized water and black fungus powder are added in a ratio of 30ml:1g, homogenized, the pH is adjusted to 4.0, 5wt% complex enzyme solution is added, enzyme hydrolysis is carried out for 2h, the temperature is then increased to 55℃ and the pH is adjusted to 6.0, 2% saccharifying enzyme is added, enzyme hydrolysis is carried out for 1h, the pH is then adjusted to 7.0, and mannase and neutral protease are added, enzyme hydrolysis is carried out at 45℃ for 3-4h, and high-temperature enzyme inactivation is carried out, to obtain an enzyme hydrolysate; the enzyme hydrolysate is centrifuged, the supernatant is taken, 3 times the amount of 95% ethanol is added, and high-speed centrifugal separation is carried out after standing for 24h, the obtained precipitate is dried to obtain black fungus polysaccharide.

[0076] The complex enzyme solution contains cellulase 15g / L and pectinase 20g / L; the mannase is added in an amount of 1g / L, and the neutral protease is added in an amount of 1.5g / L; the cellulase enzyme activity is 5000U / g, the pectinase enzyme activity is 40,000U / g; the saccharifying enzyme enzyme activity is 10,000U / g, the mannase enzyme activity is 50,000U / g, and the neutral protease enzyme activity is 60,000U / g.

[0077] (2) taking 5 g of black fungus polysaccharide and dissolving it in 200 mL of 2 mol / L sodium hydroxide solution, stirring until completely dissolved, then adding 1.5 wt% chloroacetic acid, heating to 50°C and stirring for 4 h, then cooling to room temperature to obtain a mixture, and freeze-drying the mixture into powder to obtain acidified black fungus polysaccharide;

[0078] (3) mixing the acidified black fungus polysaccharide with sodium selenite at a mass ratio of 3:1 to obtain a mixture, adding a 1% nitric acid solution to the mixture until completely dissolved, then adding BaCl2, stirring uniformly, and reacting at 70°C for 4-5 h, then adding an appropriate amount of saturated sodium sulfate solution to remove barium chloride, centrifuging to obtain the supernatant, adjusting the pH to neutral, adding 3 times the amount of 95% ethanol, standing for 24 h, and then high-speed centrifugation to obtain the precipitate, which is dried to obtain modified black fungus polysaccharide; the amount of the 1% nitric acid solution, the mixture, BaCl2, and the saturated sodium sulfate solution is 50 ml: 1 g: 0.2 g: 4 ml.

[0079] The wine processing solid by-product fermentation liquid is prepared by the following method:

[0080] a. drying the wine processing solid by-product, crushing it through an 80-mesh sieve to obtain a solid powder, adding a complex hydrolytic enzyme solution to the solid powder, and enzymatically hydrolyzing at 40°C for 8 h, then high-temperature inactivation to obtain an enzymatic hydrolysate;

[0081] b. adding Bacillus licheniformis and Lactobacillus plantarum to the enzymatic hydrolysate, adjusting the temperature to 35°C, and fermenting for 24 h, then filtering to obtain the filtrate.

[0082] The amount of the solid powder and the complex hydrolytic enzyme solution in step a is 1 g: 1.5 ml.

[0083] The complex hydrolytic enzyme solution contains cellulase 15 g / L and bromelain 20 g / L; the cellulase has an enzyme activity of 5000 U / g, and the bromelain has an enzyme activity of 100,000 U / g.

[0084] The amount of Bacillus licheniformis added in step b is 15 g / L, and the amount of Lactobacillus plantarum added is 20 g / L; the viable count of Bacillus licheniformis is 100 billion / g, and the viable count of Lactobacillus plantarum is 10 billion / g.

[0085] Comparative Example 1

[0086] A plant growth regulator for improving the stress resistance of crops, which is prepared from the following raw materials in parts by weight: black fungus polysaccharide 12 kg, wine processing solid by-product fermentation liquid 25 kg, sodium dinitrophenolate 0.08 kg, trisodium citrate 1.5 kg, gamma-aminobutyric acid 1.5 kg, and a promoter 0.4 kg; the promoter is composed of nanochitosan and sodium cocoyl glutamate at a mass ratio of 1:5.

[0087] The black fungus polysaccharide is prepared by the following method: black fungus is dried and crushed to pass through a 100-mesh sieve, deionized water and black fungus powder are added according to a ratio of 30 ml:1 g of liquid to solid, homogenized, the pH is adjusted to 4.0, 5 wt% of a complex enzyme solution is added, and enzymolysis is performed for 2 h, then the temperature is increased to 55°C and the pH is adjusted to 6.0, 2% of saccharifying enzyme is added, and enzymolysis is performed for 1 h, then the pH is adjusted to 7.0, and mannanase and neutral protease are added, and enzymolysis is performed for 3-4 h at 45°C, and the enzyme is inactivated at high temperature, to obtain an enzymolysis solution; the enzymolysis solution is centrifuged, the supernatant is taken, 3 times the amount of 95% ethanol is added, and the mixture is left to stand for 24 h, and then high-speed centrifugal separation is performed, the obtained precipitate is dried to obtain the black fungus polysaccharide;

[0088] The complex enzyme solution contains 15 g / L of cellulase and 20 g / L of pectinase; the mannanase is added in an amount of 1 g / L, and the neutral protease is added in an amount of 1.5 g / L; the cellulase has an enzyme activity of 5000 U / g, and the pectinase has an enzyme activity of 40000 U / g; the saccharifying enzyme has an enzyme activity of 10000 U / g, the mannanase has an enzyme activity of 50000 U / g, and the neutral protease has an enzyme activity of 60000 U / g.

[0089] The grape wine processing solid by-product fermentation liquid is prepared by the following method:

[0090] a. The grape wine processing solid by-product is dried and crushed to pass through an 80-mesh sieve to obtain a solid powder, a complex hydrolytic enzyme solution is added, and enzymolysis is performed for 8 h at 40°C, and the enzyme is inactivated at high temperature, to obtain an enzymolysis solution;

[0091] b. Bacillus licheniformis and Lactobacillus plantarum are added to the enzymolysis solution, the temperature is adjusted to 35°C, and fermentation is performed for 24 h, then the mixture is filtered, and the filtrate is obtained.

[0092] In step a, the ratio of the amount of the solid powder to the amount of the complex hydrolytic enzyme solution is 1 g:1.5 ml.

[0093] The complex hydrolytic enzyme solution contains 15 g / L of cellulase and 20 g / L of bromelain; the cellulase has an enzyme activity of 5000 U / g, and the bromelain has an enzyme activity of 100000 U / g.

[0094] In step b, the Bacillus licheniformis is added in an amount of 15 g / L, and the Lactobacillus plantarum is added in an amount of 20 g / L; the Bacillus licheniformis has a viable bacterial count of 100 billion / g; and the Lactobacillus plantarum has a viable bacterial count of 10 billion / g.

[0095] In the above comparative example, the raw material composition and the preparation method are basically the same as those in Example 4, and the only difference is that black fungus polysaccharide is used instead of modified black fungus polysaccharide.

[0096] Comparative Example 2

[0097] A plant growth regulator for improving stress resistance of crops, which is prepared from the following raw materials by weight: 25 kg of wine processing solid by-product fermentation liquor, 0.08 kg of sodium complex nitrophenol, 1.5 kg of trisodium citrate, 1.5 kg of gamma-aminobutyric acid, and 0.4 kg of a promoter; the promoter is composed of nanochitosan and sodium cocoyl glutamate at a mass ratio of 1:5.

[0098] The wine processing solid by-product fermentation liquor is prepared by the following method:

[0099] a. The solid powder is obtained by drying and crushing the wine processing solid by-product through an 80-mesh sieve, and a complex hydrolytic enzyme solution is added thereto for enzymolysis at 40℃ for 8h, and then high-temperature inactivation is performed to obtain an enzymolysis liquor;

[0100] b. Bacillus licheniformis and Lactobacillus plantarum are added to the enzymolysis liquor, and then the temperature is adjusted to 35℃ for fermentation for 24h, and then filtration is performed to obtain the filtrate.

[0101] The use amount ratio of the solid powder to the complex hydrolytic enzyme solution in step a is 1g:1.5ml.

[0102] The complex hydrolytic enzyme solution contains cellulase 15g / L and bromelain 20g / L; the cellulase has an enzyme activity of 5000U / g, and the bromelain has an enzyme activity of 100,000U / g.

[0103] The addition amount of Bacillus licheniformis in step b is 15g / L, and the addition amount of Lactobacillus plantarum is 20g / L; the viable count of Bacillus licheniformis is 100 billion / g, and the viable count of Lactobacillus plantarum is 10 billion / g.

[0104] The raw material composition and preparation method in the above comparative example are basically the same as those in Example 4, and the only difference is that the modified Auricularia auricular polysaccharide is not contained.

[0105] Comparative Example 3

[0106] A plant growth regulator for improving stress resistance of crops, which is prepared from the following raw materials by weight: 12 kg of modified Auricularia auricular polysaccharide, 0.08 kg of sodium complex nitrophenol, 1.5 kg of trisodium citrate, 1.5 kg of gamma-aminobutyric acid, and 0.4 kg of a promoter; the promoter is composed of nanochitosan and sodium cocoyl glutamate at a mass ratio of 1:5.

[0107] The modified Auricularia auricular polysaccharide is prepared by the following method:

[0108] (1) Take dried black fungus, crush it through a 100 mesh sieve, and add deionized water and black fungus powder according to a ratio of 30 ml:1 g of liquid to material. Homogenize, adjust the pH to 4.0, and add a 5 wt% complex enzyme solution. Enzymatically hydrolyze for 2 h, then raise the temperature to 55°C and adjust the pH to 6.0. Add 2% saccharifying enzyme, enzymatically hydrolyze for 1 h, then adjust the pH to 7.0, add mannanase and neutral protease, and enzymatically hydrolyze at 45°C for 3-4 h. Inactivate the enzymes at high temperature, and obtain an enzymatic hydrolysate. Centrifuge the enzymatic hydrolysate, take the supernatant, add 3 times the amount of 95% ethanol, and centrifuge at high speed after standing for 24 h. Dry the precipitate obtained to obtain black fungus polysaccharide;

[0109] The complex enzyme solution contains 15 g / L of cellulase and 20 g / L of pectinase. The mannanase is added in an amount of 1 g / L, and the neutral protease is added in an amount of 1.5 g / L. The cellulase has an enzyme activity of 5000 U / g, and the pectinase has an enzyme activity of 40000 U / g. The saccharifying enzyme has an enzyme activity of 10000 U / g, the mannanase has an enzyme activity of 50000 U / g, and the neutral protease has an enzyme activity of 60000 U / g.

[0110] (2) Take 5 g of black fungus polysaccharide, dissolve it in 200 mL of 2 mol / L sodium hydroxide solution, stir until dissolved, then add 1.5 wt% chloroacetic acid, heat to 50°C, and stir to react for 4 h. After cooling to room temperature, obtain a mixed solution, freeze-dry the mixed solution to obtain acidified black fungus polysaccharide powder.

[0111] (3) Mix the acidified black fungus polysaccharide with sodium selenite according to a mass ratio of 3:1 to obtain a mixture. Add a 1% nitric acid solution to the mixture until dissolved, then add BaCl2, stir until uniform, and react in a 70°C water bath for 4-5 h. After the reaction is completed, add an appropriate amount of saturated sodium sulfate solution to remove barium chloride, centrifuge to separate, adjust the pH to neutral, add 3 times the amount of 95% ethanol, and centrifuge at high speed after standing for 24 h. Dry the precipitate obtained to obtain modified black fungus polysaccharide. The amount ratio of the 1% nitric acid solution, the mixture, BaCl2, and the saturated sodium sulfate solution is 50 ml:1 g:0.2 g:4 ml.

[0112] The raw material composition and preparation method in the above comparative example are basically the same as in Example 4, except that the fermentation broth of the solid by-product of grape wine processing is not contained.

[0113] Comparative Example 4

[0114] A plant growth regulator for improving the stress resistance of crops, which is made from the following raw materials in parts by weight: modified black fungus polysaccharide 12 kg, fermentation broth of solid by-product of grape wine processing 25 kg, sodium nitrofuroate 0.08 kg, trisodium citrate 1.5 kg, and γ-aminobutyric acid 1.5 kg.

[0115] The modified Auricularia auricular polysaccharide is prepared by the following method:

[0116] (1) After drying Auricularia auricular, it is crushed to pass through a 100-mesh sieve, and deionized water and Auricularia auricular powder are added at a ratio of 30 ml:1 g of liquid to solid, and homogenized. The pH is adjusted to 4.0, and 5 wt% of a complex enzyme solution is added. After 2 h of enzymatic hydrolysis, the temperature is increased to 55°C and the pH is adjusted to 6.0. Then 2% of saccharifying enzyme is added, and the pH is adjusted to 7.0. After 1 h of enzymatic hydrolysis, mannase and neutral protease are added, and the enzymatic hydrolysis is carried out at 45°C for 3-4 h. The enzyme is inactivated at high temperature, and an enzyme hydrolysate is obtained. The enzyme hydrolysate is centrifuged to separate the supernatant. The supernatant is added with 3 times the amount of 95% ethanol, and is left to stand for 24 h. Then high-speed centrifugal separation is carried out, and the obtained precipitate is dried to obtain Auricularia auricular polysaccharide.

[0117] The complex enzyme solution contains 15 g / L of cellulase and 20 g / L of pectinase. The addition amount of the mannase is 1 g / L, and the addition amount of the neutral protease is 1.5 g / L. The enzyme activity of the cellulase is 5000 U / g, and the enzyme activity of the pectinase is 40000 U / g. The enzyme activity of the saccharifying enzyme is 10000 U / g, the enzyme activity of the mannase is 50000 U / g, and the enzyme activity of the neutral protease is 60000 U / g.

[0118] (2) 5 g of Auricularia auricular polysaccharide is dissolved in 200 mL of 2 mol / L sodium hydroxide solution, and stirred until completely dissolved. Then 1.5 wt% chloroacetic acid is added, heated to 50°C, and stirred for 4 h. After cooling to room temperature, a mixed solution is obtained. The mixed solution is freeze-dried into powder to obtain acidified Auricularia auricular polysaccharide.

[0119] (3) The acidified Auricularia auricular polysaccharide is mixed with sodium selenite at a mass ratio of 3:1 to obtain a mixture. The mixture is dissolved completely by adding 1% nitric acid solution, and then BaCl2 is added. After stirring uniformly, the reaction is carried out at 70°C water bath for 4-5 h. After the reaction is completed, a proper amount of saturated sodium sulfate solution is added to remove barium chloride. After centrifugal separation, the supernatant is obtained, and the pH value is adjusted to neutral. After standing for 24 h with 3 times the amount of 95% ethanol, high-speed centrifugal separation is carried out. The obtained precipitate is dried to obtain modified Auricularia auricular polysaccharide. The amount ratio of the 1% nitric acid solution, the mixture, BaCl2, and the saturated sodium sulfate solution is 50 ml:1 g:0.2 g:4 ml.

[0120] The grape wine processing solid by-product fermentation liquid is prepared by the following method:

[0121] a. The grape wine processing solid by-product is dried and crushed to pass through an 80-mesh sieve to obtain a solid powder. A complex hydrolytic enzyme solution is added, and the enzymatic hydrolysis is carried out at 40°C for 8 h. After high-temperature inactivation, an enzyme hydrolysate is obtained.

[0122] b. After adding bacillus licheniformis and lactobacillus plantarum to the enzymatic hydrolysis solution, the temperature was adjusted to 35℃, and fermentation was carried out for 24 hours, then filtration was performed, and the filtrate was obtained.

[0123] The ratio of the solid powder to the complex hydrolytic enzyme solution in step a is 1g:1.5ml.

[0124] The complex hydrolytic enzyme solution contains cellulase 15g / L and bromelain 20g / L; the cellulase enzyme activity is 5000U / g, and the bromelain enzyme activity is 100,000U / g.

[0125] The bacillus licheniformis is added in an amount of 15g / L, and the lactobacillus plantarum is added in an amount of 20g / L; the bacillus licheniformis has a viable bacterial count of 100 billion / g; and the lactobacillus plantarum has a viable bacterial count of 10 billion / g.

[0126] The raw material composition and preparation method in the above comparative example are basically the same as those in example 4, and the only difference is that no accelerator is contained.

[0127] Test Example 1

[0128] Effect of the present application on improving the salt and alkali stress resistance of small rape seedlings

[0129] The test was carried out by using a greenhouse potting method, and the test site was an agricultural planting base in Liaocheng City. Small rape seeds were sown in hole pots mixed with 1:1 of nutrient soil and vermiculite, and after two weeks of cultivation in a culture room at a temperature of 25℃, small rape seedlings were obtained. The rape seedlings with consistent growth vigor were transplanted into pots containing 500g of test soil (alkali-hydrolyzable nitrogen content 88.55mg / kg, available phosphorus content 37.23mg / kg, available potassium content 107.62mg / kg, organic matter content 20.19g / kg, pH 6.55), and the potting soil was fertilized at a level of N-P2O5-K2O of 20-10-20. One plant was transplanted in each pot. After transplantation, each pot was watered with 200mL of water, and then each pot was watered with 100mL of water every other day. Twenty days after the seedlings were transplanted, the treatment was started, and the test was set up with 9 treatment groups, of which groups 1-4 were example 1-4 treatment groups, groups 5-8 were comparative example 1-4 treatment groups, and group 9 was a blank control group. The specific treatment method of each treatment group was as follows: 100ml of salt and alkali liquid + 0.15g of different plant growth regulators obtained from the treatment groups were used to irrigate the roots on the 20th day after the seedlings were transplanted, wherein the 100ml of salt and alkali liquid was composed of 50ml of 0.1mol / L NaCl + 50ml of 0.15mol / L NaHCO3 solution, and 0.15g of deionized water was used to replace the blank control group. After 10 days of irrigation, the plant height, fresh weight, dry weight, and chlorophyll content of each treatment group were measured and statistically analyzed, and the average value was taken from 4 repeated tests. The test results are shown in Table 1.

[0130] Table 1 Test results of different treatment groups of Brassica rapa L. resisting salt and alkali stress

[0131] Plant height / cm Chlorophyll content mg / g Fresh weight / g Dry weight / g Example 1 5.5 30.6 4.46 1.47 Example 2 5.7 30.9 4.32 1.35 Example 3 5.6 30.1 4.23 1.39 Example 4 5.7 30.8 4.29 1.37 Comparative Example 1 4.7 25.3 3.95 1.19 Comparative Example 2 4.4 24.1 3.22 0.99 Comparative Example 3 4.5 25.5 2.95 0.91 Comparative Example 4 5.1 27.9 4.16 1.26 Blank control 3.8 22.1 1.96 0.63

[0132] As can be seen from the results of Table 1 above, compared with the blank control group, the salt and alkali stress relief effect of Brassica rapa L. seedlings treated with the plant growth regulator prepared by Examples 1-4 of the present application is significantly better than that of Comparative Examples 1-4, which shows that the modified Auricularia auricular polysaccharide used in the raw material composition of the present application is seleniumized Auricularia auricular polysaccharide, and the introduction of selenium element improves the biological activity of Auricularia auricular polysaccharide, and significantly promotes the growth of plants, and after being used in combination with grape wine processing solid by-product fermentation liquor, the two have a synergistic effect, which significantly improves the salt and alkali resistance of crops.

[0133] Test Example 2

[0134] Effect of the present application on improving the low temperature cold damage resistance of strawberries

[0135] The test site is located in Liaocheng City, Shandong Province, and a pot experiment is carried out. The test crop is 2-leaf red strawberry, and the pot test soil is collected from the 0-20 cm plough layer soil of the local representative dry land. The size of the pot is 10 cm x 12 cm, and the soil is 500 g. The fertilization level of the potting soil (N-P2O5-K2O) is 20-10-15. The 2-leaf red strawberry seedlings with consistent growth are transplanted into the above pots, one plant per pot. Different treatments are started 15 days after transplantation. The test sets 9 treatment groups and numbers them as 1-9. The pots of groups 1-9 are placed in a 4℃ incubator for culture, and different test solutions are sprayed / rooted at the same time. Groups 1-4 correspond to the plant growth regulators obtained by Examples 1-4, respectively, and groups 5-8 correspond to the plant growth regulators obtained by Comparative Examples 1-4, respectively. The usage and dosage is 0.15 g / pot, which can be directly sprayed or sprayed after diluting with an appropriate amount of water. The spraying frequency is 5 days / time. The 9th group is the blank control group, and the same amount of water is sprayed each time. The growth conditions of the strawberries are consistent with those of the other treatment groups. After spraying for 15 days, the growth of the strawberries is observed, and the plant height of each treatment group is measured. Then the strawberry plants are killed in a 105℃ oven for 30 min, dried at 75℃ to constant weight, and the above-ground biomass and root biomass are measured. The test is repeated 4 times to take the average value, and the specific test results are shown in Table 2.

[0136] Table 2 Test results of different treatment groups of strawberries under low temperature cold stress

[0137] Group 1 Group 2 Group 3 Group 4 Group 5 Group 6 Group 7 Group 8 Group 9 Aboveground biomass / g 20.4 19.4 19.9 19.5 14.3 13.6 13.1 17.5 9.8 Underground biomass / g 4.9 4.7 4.8 4.7 3.6 3.5 3.2 4.2 2.4 Plant height / cm 20.6 19.8 20.5 19.9 17.9 15.9 15.1 18.2 12.9

[0138] From the data of Table 2, it can be seen that the plant height, aboveground biomass and underground biomass of groups 1-4 are obviously higher than those of the blank control group 9, and are also better than those of the comparative examples 1-4, which indicates that the plant growth regulator for improving stress resistance of crops of the present application can obviously promote the growth and development of strawberries under low temperature conditions, and significantly improve the ability of strawberries to resist low-temperature cold damage.

[0139] Test Example 3

[0140] Effect of different compositions of the present application on the drought stress resistance of Brassica rapa L. seedlings

[0141] The test was carried out by using a greenhouse potting method, and the test site was an agricultural planting base in Liaocheng City. Brassica rapa L. seeds were sown in hole pots mixed with nutrient soil and vermiculite at a ratio of 1:1, and after two weeks of cultivation in a culture room at a temperature of 25°C, Brassica rapa L. seedlings were obtained. The Brassica rapa L. seedlings with consistent growth vigor were transplanted into pots containing 500g of test soil (alkali-hydrolyzable nitrogen content 88.55mg / kg, available phosphorus content 37.23mg / kg, available potassium content 107.62mg / kg, organic matter content 20.19g / kg, pH 6.55) (length 10cm x 10cm x height 12cm), and the fertilization level of the potting soil (N-P2O5-K2O) was 20-10-20. One plant was transplanted in each pot. Ten days after the seedlings were transplanted, moderate drought treatment was started, and six treatment groups were set up, of which group 1 was a blank control group, i.e. a moderate drought treatment group, and the soil water content was 45%-55% of the soil mass. The consumed water was calculated by weighing every other day and the corresponding amount of clean water was added to maintain the soil water content. Groups 2-6 were treated with plant growth regulators 1-5, respectively, i.e. after moderate drought treatment (same as the treatment method of group 1), plant growth regulators 1-5 solutions were sprayed / rooted with a usage of 0.15g / pot. The plant growth regulator 1 was the product obtained in Example 4; the plant growth regulator 2 was basically the same as Example 4, except that the promoter was composed of nano-chitosan and sodium cocoyl glutamate di-sodium in a mass ratio of 1:1; the plant growth regulator 3 was basically the same as Example 4, except that the promoter was composed of nano-chitosan and sodium cocoyl glutamate di-sodium in a mass ratio of 1:3; the plant growth regulator 4 was basically the same as Example 4, except that the promoter was composed of nano-chitosan and sodium cocoyl glutamate di-sodium in a mass ratio of 1:7; the plant growth regulator 5 was basically the same as Example 4, except that the promoter was composed of nano-chitosan and sodium cocoyl glutamate di-sodium in a mass ratio of 1:9. After 10 days of spraying and irrigation, the plant height, fresh weight, dry weight and chlorophyll content of each treatment group were measured and statistically analyzed, and the test was repeated four times to take the average value. The test results are shown in Table 3.

[0142] Effect of different compositions of the present application on the drought stress resistance of Brassica rapa L. seedlings

[0143] Plant height / cm Chlorophyll content mg / g Fresh weight / g Dry weight / g Group 1 3.5 21.5 2.01 0.59 Group 2 5.5 29.8 4.29 1.29 Group 3 3.7 24.9 2.85 0.86 Group 4 4.2 27.6 3.21 0.97 Group 5 4.3 26.2 3.85 1.16 Group 6 3.9 23.9 3.67 0.11

[0144] This invention also determined the activity of defense enzymes in rapeseed seedling leaves from the above-mentioned different treatment groups. The specific procedure was as follows: 0.1g of rapeseed leaves were weighed from each treatment group, 3ml of PBS (pH 7.8) was added, and the mixture was homogenized in an ice bath. Then, 2ml of PBS was added to rinse the mortar, and the mixture was poured into centrifuge tubes and centrifuged at 10000r / min for 20min at 4℃. The supernatant was the crude enzyme solution, which could be used for enzyme activity determination. The contents of SOD, CAT, and POD in the crude enzyme extract were determined separately. This experiment was repeated four times, and the average value was taken. Superoxide dismutase (SOD) was determined using the nitroblue tetrazolium photoreduction method; catalase (CAT) and peroxidase (POD) were determined using ultraviolet spectrophotometry. Specific results are as follows: Figure 1-3 As shown above. Figure 1-3 The results show that the composition ratio of the promoter of the present invention has a significant effect on the activities of SOD, CAT and POD enzymes in rapeseed seedlings. Only the promoter composed of the proportion of the present invention has the best effect on improving the activity of defensive enzymes in rapeseed seedlings. This is because the promoter of the present invention promotes the penetration and absorption of various active substances, optimizes the nutrient distribution and physiological metabolic processes in the plant, thereby improving the plant's stress resistance and yield.

[0145] It should be noted that the above embodiments are merely some preferred embodiments of the present invention, and not all embodiments. Obviously, based on the above embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort should fall within the scope of protection of the present invention.

Claims

1. A plant growth regulator for improving crop stress resistance, characterized in that, It is made from the following weight parts of raw materials: modified black fungus polysaccharide 10-20 parts, grape wine processing solid by-product fermentation liquor 30-50 parts, sodium complex nitrophenol 0.05-0.15 parts, trisodium citrate 1-3 parts, gamma-aminobutyric acid 1-3 parts, and promoter 0.3-0.6 parts; the promoter is nano-chitosan and sodium cocoyl glutamate with a mass ratio of 1:5; The modified black fungus polysaccharide is prepared by the following method: (1) After drying, the black fungus is crushed to pass through a 100-mesh sieve, deionized water and black fungus powder are added according to the solid-liquid ratio, homogenized, the pH is adjusted to 4.0-4.5, then 5wt% complex enzyme solution is added, and enzyme hydrolysis is carried out for 2h, then the temperature is increased to 55℃ and the pH is adjusted to 6.0-6.5, 2% saccharifying enzyme is added, and enzyme hydrolysis is carried out for 1h, then the pH is adjusted to 6.5-7.0, and mannase and neutral protease are added, and enzyme hydrolysis is carried out at 40-45℃ for 3-4h, and high-temperature enzyme inactivation is carried out, to obtain an enzyme hydrolysate; the enzyme hydrolysate is centrifuged, the supernatant is taken, 3 times the amount of 95% ethanol is added, and high-speed centrifugal separation is carried out after standing for 24h, and the obtained precipitate is dried to obtain black fungus polysaccharide; (2) 5g of black fungus polysaccharide is dissolved in 200mL of 2mol / L sodium hydroxide solution, stirred until dissolved, then 1.5wt% chloroacetic acid is added, heated to 50℃ and stirred for 4h, then cooled to room temperature to obtain a mixed solution, and the mixed solution is freeze-dried into powder to obtain acidified black fungus polysaccharide; (3) The acidified black fungus polysaccharide and sodium selenite are mixed in proportion to obtain a mixture, 1% nitric acid solution is added to the mixture until dissolved, then BaCl2 is added, stirred uniformly, and reacted in a 70℃ water bath for 4-5h, then saturated sodium sulfate solution is added to remove BaCl2, centrifuged, the supernatant is taken, the pH is adjusted to neutral, 3 times the amount of 95% ethanol is added, and high-speed centrifugal separation is carried out after standing for 24h, and the obtained precipitate is dried to obtain modified black fungus polysaccharide; The stress resistance is the ability to resist salt-alkali, drought, and low temperature stress; In step (1), the solid-liquid ratio of black fungus powder to deionized water is 1g:30ml; the complex enzyme solution contains 10-15g / L of cellulase and 15-20g / L of pectinase; the addition amount of mannase is 0.5-1g / L, and the addition amount of neutral protease is 1.5-2g / L; In step (3), the mass ratio of acidified black fungus polysaccharide to sodium selenite is 3:1, and the ratio of the amount of 1% nitric acid solution, the mixture, BaCl2, and saturated sodium sulfate solution is 50ml:1g:0.2g:3-5ml; The grape wine processing solid by-product fermentation liquor is prepared by the following method: a. The grape wine processing solid by-product is dried and crushed to pass through an 80-mesh sieve to obtain a solid powder, and a complex hydrolysis enzyme solution is added to the solid powder for enzyme hydrolysis at 35-40℃ for 6-8h, and high-temperature inactivation is carried out, to obtain an enzyme hydrolysate; the complex hydrolysis enzyme solution contains 10-15g / L of cellulase and 20-25g / L of bromelain; the cellulase has an enzyme activity of 5000U / g, and the bromelain has an enzyme activity of 100,000U / g; b. After adding bacillus licheniformis and lactobacillus plantarum into the enzymatic hydrolysate, the temperature is adjusted to 30-35 DEG C, and fermentation is carried out for 24 hours, then filtration is carried out, and the filtrate is obtained.

2. The plant growth regulator for increasing stress resistance of crops according to claim 1, characterized by, The enzyme activity of the cellulase is 5000 U / g, the enzyme activity of the pectinase is 40000 U / g; the enzyme activity of the saccharifying enzyme is 10000 U / g, the enzyme activity of the mannanase is 50000 U / g, and the enzyme activity of the neutral protease is 60000 U / g.

3. The plant growth regulator for increasing stress resistance of crops according to claim 1, characterized by, The ratio of the solid powder to the complex hydrolytic enzyme solution in step a is 1g:1.5-2ml.

4. The plant growth regulator for increasing stress resistance of crops according to claim 1, characterized by, The bacillus licheniformis is added in an amount of 10-15g / L, and the lactobacillus plantarum is added in an amount of 20-30g / L.

5. The plant growth regulator for increasing stress resistance of crops according to claim 1, characterized by, The viable count of the bacillus licheniformis is 100 billion / g, and the viable count of the lactobacillus plantarum is 10 billion / g.

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