CN112021027B - Method for improving stress resistance of plants and promoting flower bud differentiation - Google Patents
Method for improving stress resistance of plants and promoting flower bud differentiation Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
- A01G17/005—Cultivation methods
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
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- A01N43/06—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings
- A01N43/12—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom five-membered rings condensed with a carbocyclic ring
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/34—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom
- A01N43/36—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom five-membered rings
- A01N43/38—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one nitrogen atom as the only ring hetero atom five-membered rings condensed with carbocyclic rings
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Abstract
The invention relates to a method for improving the stress resistance of plants and promoting flower bud differentiation, which is characterized in that after fruits are picked in autumn, nutrient solution containing chitooligosaccharide peptide magnesium is sprayed on leaf surfaces.
Description
Technical Field
The present invention relates to a method for improving stress resistance of a plant and promoting flower bud differentiation.
Background
After fruit picking of fruit trees in autumn, in order to ensure the yield of the next year, fertilization and nutrition supplementation are generally carried out, commonly called as 'monthly manure', because fruits of the fruit trees need to absorb a large amount of nutrition from growth to maturity, and nutrient substances in the fruit trees are basically consumed completely after the fruit trees are picked, so that timely supplementation is needed.
At present, nutrient substances of trees are supplemented, usually a basal fertilizer application mode is adopted for nutrient supplement, and basal fertilizer is conveyed into the trees through root systems. The roots are mainly pulled by the transpiration to absorb nutrients, however, the temperature is reduced after autumn, the transpiration of leaves is reduced, the activity of the roots is reduced, slow absorption of the nutrients is caused, and fertilizer waste is easily caused.
By spraying on the leaf surface, the temperature in autumn is low, the leaf begins to turn yellow and fall off under the influence of the temperature, although the fall off of the leaf can be prevented by supplementing auxin and cytokinin, the growth and division of the cell are energy consumption, and the energy is also a burden to the tree body.
The chlorophyll molecule contains a "head" of a porphyrin ring and a "tail" of phytol. The magnesium atom is centered in the porphyrin ring and is biased to a positive charge, and the nitrogen atom associated with it is biased to a negative charge, so that the porphyrin is polar, hydrophilic and can bind to proteins. The magnesium atom in the porphyrin ring may be replaced by a hydrogen ion. Abscisic acid is secreted in plants in autumn, and hydrogen ions in the abscisic acid replace magnesium atoms to form pheophytin, so that leaves are brown, green and abscission are avoided, and photosynthesis of the leaves is influenced.
Chitosan oligosaccharide has good water solubility and is easy to absorb, and can effectively supplement nutrition to trees, but the ortho position of the free amino group of the chitosan oligosaccharide is hydroxyl, has the function of chelating divalent metal ions, is a high-performance metal ion trapping agent, reacts with the divalent metal ions to generate water-insoluble macromolecular chelating salt, and forms flocculent precipitate, so that the chitosan oligosaccharide and magnesium ions do not coexist in neutral environment, usually need to coexist in acidic environment with pH lower than 5, but adopt chitosan oligosaccharide magnesium with pH lower than 5 for treatment, and because magnesium in chlorophyll is easily replaced by hydrogen ions in the acidity, although nutrition is supplemented, leaves still lose green, and photosynthesis is influenced.
At present, no method for coexistence of chitosan oligosaccharide and magnesium ions in a neutral environment exists, and no method for better improving plant stress resistance and promoting flower bud differentiation exists.
Disclosure of Invention
The invention provides a method for improving the stress resistance of plants and promoting flower bud differentiation, and solves the technical problems of 1) a method for improving the stress resistance of plants and promoting flower bud differentiation; 2) the quality of the product is unstable by using cellulase; 3) hydrogen peroxide is used for hydrolysis, and hydrolysis products are uncontrollable, so that waste is caused to chitosan; 4) chitosan oligosaccharide and magnesium can coexist only in an acid environment, but the acid environment is not favorable for the problem that the magnesium supplement effect is not satisfactory, the pH is higher than 6, and the chitosan oligosaccharide and the magnesium do not coexist.
In order to solve the technical problems, the invention adopts the following technical scheme:
a method for improving plant stress resistance and promoting flower bud differentiation comprises spraying chitooligosaccharide magnesium diluent to leaf surface within 10 days after picking fruit in autumn.
The chitooligosaccharide peptide magnesium also comprises one or two of auxin or cytokinin.
The pH value of the chitooligosaccharide peptide magnesium is 6.0-7.0.
The chitooligosaccharide peptide magnesium is prepared according to the following steps:
carrying out enzymolysis on chitosan in an acid environment by lysozyme to obtain agricultural chitosan oligosaccharide enzymolysis liquid;
heating the chitosan oligosaccharide enzymolysis liquid under certain conditions;
adding alkali for hydrolysis to obtain chitosan oligosaccharide peptide hydrolysate;
adding magnesium ions to obtain liquid chitooligosaccharide magnesium; or the like, or, alternatively,
drying the liquid chitooligosaccharide peptide magnesium to obtain chitooligosaccharide peptide magnesium; or
Mixing one or two of auxin and cytokinin with liquid chitooligosaccharide peptide magnesium or chitooligosaccharide peptide magnesium to obtain chitooligosaccharide peptide magnesium mixture.
The acid environment comprises water, water-soluble unsaturated dibasic organic acid or water-soluble unsaturated dibasic organic acid anhydride and ammonium ions;
the pH value of the acidic environment is 4.0-5.5;
the enzymolysis is carried out for 6-48 h at the temperature of 30-60 ℃;
the mass ratio of the chitosan to the acid environment to the lysozyme is 1-10: 89.9-98.9: 0.1-1;
the mass ratio of water, water-soluble unsaturated dibasic organic acid or water-soluble unsaturated dibasic organic acid anhydride and ammonium ions in the acidic environment is 60-92: 6.5-39: 1-8.
The certain conditions are that the pressure is 3 atm-45 atm, the heating temperature is 130-280 ℃, and the heating time is 20-60 min;
the mass ratio of the alkali to the chitosan oligosaccharide enzymolysis liquid is 2-40: 60-98; the hydrolysis time is 2-30 min;
the alkali is one or more of sodium hydroxide, potassium hydroxide and their water solution;
the molar ratio of the added magnesium ions to the water-soluble unsaturated dibasic organic acid or the water-soluble unsaturated dibasic organic acid anhydride in the agricultural chitooligosaccharide peptide hydrolysate is less than 1.
The blending includes dissolving and mixing.
Diluting the chitooligosaccharide peptide magnesium to obtain a diluent; or the like, or, alternatively,
diluting the chitooligosaccharide peptide magnesium mixture to obtain chitooligosaccharide peptide magnesium diluent.
The chitooligosaccharide peptide magnesium is diluted by water, and the EC value of the diluted solution is lower than 4 mS/cm;
the chitooligosaccharide peptide magnesium mixture is diluted by water until the content of auxin or cytokinin in the diluted solution is 100 mg/L-150 mg/L, and the EC value of the diluted solution is lower than 4 mS/cm.
The invention has the following beneficial technical effects:
1. the application has neutral pH, can coexist magnesium and chitosan oligosaccharide, has a good supplement effect on magnesium, participates in the assimilation reaction of carbon dioxide in chloroplast, improves the photosynthesis rate, and is favorable for the synthesis of sugar and starch.
2. In the application, auxin or cytokinin is added to spray the leaf surfaces, auxin supplement is carried out on the far shaft ends of the abscission layers to prevent the leaves from falling off, meanwhile, the falling-off of the leaves is the basis for improving the stress resistance of the plants and promoting the flower bud differentiation, the existence of the leaves provides guarantee for the absorption of chitosan oligosaccharide by the tree body, the promotion of the auxin is energy consumption, the chitosan oligosaccharide supplements the energy consumed by the growth, the chitosan oligosaccharide and the energy play a mutual promotion role, in addition, the chitosan oligosaccharide magnesium enters the tree body through the leaves, not only can supplement nutrition for the tree body, but also is beneficial to the synthesis of sugar and starch, the increase of sugar in the tree body can resist the protoplasm of cells from being damaged by low temperature, simultaneously, plants are induced to generate low-temperature photosynthetic factors, the lignification degree of the plants is improved, the cold resistance is increased, and the nutrition accumulated at the action rate is improved through the supplemented nutrition and the magnesium, and sufficient nutrition guarantee is provided for flower bud differentiation in the next year.
3. The chitosan oligosaccharide quality of this application preparation is stable, and this application utilizes lysozyme to destroy beta-1, 4 glycosidic bond between the chitosan, makes the chitosan enzymolysis, and lysozyme in this application is a single enzyme, and the stable quality does not need the cooperation between the various enzymes, consequently, can make the chitosan oligosaccharide quality of this application more stable.
4. The enzymolysis product is controllable, lysozyme only destroys beta-1, 4 glycosidic bonds between chitosan, and the lysozyme can not oxidize beta-1, 4 glycosidic bonds and hydroxyl groups like hydrogen peroxide, so that the quality of the final product is unstable, and even water-insoluble substances are generated, and the chitosan is required to be filtered, thereby causing waste.
5. The water-soluble unsaturated dibasic organic acid or the water-soluble unsaturated dibasic organic acid anhydride reacts with ammonium ions to generate, unsaturated amic acid, under certain pressure and temperature, the unsaturated amic acid is thermally polymerized to generate polyimide, the polyimide is hydrolyzed to obtain polyamino acid salt, the polyamino acid salt has better chelation property, and is chelated with metal cations, so that no precipitate can be generated, meanwhile, the metal cations and the chitosan oligosaccharide can be prevented from reacting to generate precipitate, and the compatibility of the chitosan oligosaccharide and the fertilizer is improved.
6. Chitosan oligosaccharide's film forming ability receives the influence of pH, pH is lower the film forming ability worse, this application pH is neutral partially, its film forming ability is better than the chitosan oligosaccharide that pH is less than 5, this application spouts chitooligosaccharide peptide magnesium in the blade surface, the not absorbed chitooligosaccharide peptide magnesium can form a rete on the plant leaf, moisture for reducing plant transpiration and taking away, reduce the consumption of trees moisture, thereby prolong the detention time of leaf on the tree, the blade of detention passes through photosynthesis, for plant reserve nutrition, improve plant resistance and promote the flower bud differentiation.
Detailed Description
The present invention is further illustrated by the following specific examples.
Example 1
A method for improving plant stress resistance and promoting flower bud differentiation comprises spraying chitooligosaccharide magnesium diluent on leaf surface in 3 days after fruit picking in autumn.
The pH of the magnesium chitooligosaccharide peptide is 6.5.
The preparation method of the chitooligosaccharide peptide magnesium comprises the following steps:
performing enzymolysis on chitosan in an acidic environment for 12 hours by lysozyme at the temperature of 45 ℃ to obtain chitosan oligosaccharide enzymolysis liquid;
the acid environment is obtained by mixing deionized water, maleic acid and ammonium bicarbonate according to the mass ratio of 62:20:18, adding 5% of sodium hydroxide after mixing, and adjusting the pH value to 4.5;
the mass ratio of the chitosan, the acidic environment and the lysozyme is 6:93.7: 0.3.
Reacting for 30min under the conditions that the pressure is 11atm and the temperature is 180 ℃;
adding potassium hydroxide, and hydrolyzing to obtain chitosan oligosaccharide peptide hydrolysate for agriculture;
the mass ratio of the potassium hydroxide to the chitosan oligosaccharide enzymolysis liquid is 2: 98; the hydrolysis time is 10 min;
adding magnesium nitrate into the agricultural oligo chitose peptide hydrolysate to obtain liquid oligo chitose peptide magnesium;
the molar ratio of magnesium ions to maleic acid in the magnesium nitrate was 0.8.
Diluting the liquid chitooligosaccharide peptide magnesium by 100 times by adopting water to obtain chitooligosaccharide peptide magnesium diluent.
Example 2
A method for improving plant stress resistance and promoting flower bud differentiation comprises spraying chitooligosaccharide magnesium diluent on leaf surface in 3 days after fruit picking in autumn.
The pH of the chitooligosaccharide peptide magnesium is 6.5, and the chitooligosaccharide peptide magnesium also comprises gibberellin, wherein the mass ratio of the chitooligosaccharide peptide magnesium to the gibberellin is 90: 10.
The preparation method of the chitooligosaccharide peptide magnesium comprises the following steps:
performing enzymolysis on chitosan in an acidic environment for 12 hours by lysozyme at the temperature of 45 ℃ to obtain chitosan oligosaccharide enzymolysis liquid;
the acid environment is obtained by mixing deionized water, maleic acid and ammonium bicarbonate according to the mass ratio of 62:20:18, adding 5% of sodium hydroxide after mixing, and adjusting the pH value to 4.5;
the mass ratio of the chitosan, the acidic environment and the lysozyme is 6:93.7: 0.3.
Reacting for 30min under the conditions that the pressure is 11atm and the temperature is 180 ℃;
adding potassium hydroxide, and hydrolyzing to obtain chitosan oligosaccharide peptide hydrolysate for agriculture;
the mass ratio of the potassium hydroxide to the chitosan oligosaccharide enzymolysis liquid is 2: 98; the hydrolysis time is 10 min;
adding magnesium nitrate into the agricultural oligo chitose peptide hydrolysate to obtain liquid oligo chitose peptide magnesium;
the molar ratio of magnesium ions to maleic acid in the magnesium nitrate is 0.8;
adding gibberellin into the liquid chitooligosaccharide peptide magnesium to obtain a liquid chitooligosaccharide peptide magnesium mixture.
Diluting the liquid chitooligosaccharide peptide magnesium mixture by 100 times by water, wherein the gibberellin content in the diluent is 100mg/L, and obtaining chitooligosaccharide peptide magnesium diluent.
Example 3
A method for improving plant stress resistance and promoting flower bud differentiation comprises spraying chitooligosaccharide magnesium diluent on leaf surface 5 days after fruit picking in autumn.
The pH of the chitooligosaccharide peptide magnesium is 6.2, and the chitooligosaccharide peptide magnesium also comprises gibberellin, wherein the mass ratio of the chitooligosaccharide peptide magnesium to the gibberellin is 50: 50.
The preparation method of the chitooligosaccharide peptide magnesium comprises the following steps:
performing enzymolysis on chitosan for 26 hours by lysozyme in an acid environment at the temperature of 30 ℃ to obtain agricultural chitosan oligosaccharide enzymolysis liquid;
the acid environment is obtained by mixing deionized water, pentenoic acid anhydride and ammonium sulfate according to the mass ratio of 86:8:6, adding 5% of sodium hydroxide after mixing, and adjusting the pH value to 4.2;
the acid environment is a composition of deionized water, pentenoic acid anhydride and ammonium sulfate according to a mass ratio of 86:8: 6;
the mass ratio of the chitosan, the acidic environment and the lysozyme is 8:91.6: 0.4.
Reacting for 20min under the conditions that the pressure is 28atm and the temperature is 230 ℃;
adding 8% potassium hydroxide water solution, and hydrolyzing to obtain agricultural chitooligosaccharide peptide hydrolysate;
the mass ratio of the 8 percent potassium hydroxide aqueous solution to the chitosan oligosaccharide enzymolysis liquid is 25: 75; the hydrolysis time was 3 min;
adding magnesium nitrate into the agricultural oligo chitose peptide hydrolysate to obtain liquid oligo chitose peptide magnesium;
the molar ratio of magnesium ions to maleic acid in the magnesium nitrate was 0.6.
Adding gibberellin into the liquid chitooligosaccharide peptide magnesium to obtain a liquid chitooligosaccharide peptide magnesium mixture.
Diluting the liquid chitooligosaccharide peptide magnesium mixture by water until the gibberellin content in the diluent is 150mg/L, thus obtaining chitooligosaccharide peptide magnesium diluent.
Example 4
A method for improving plant stress resistance and promoting flower bud differentiation comprises spraying chitooligosaccharide magnesium diluent on leaf surface 8 days after fruit picking in autumn.
The pH of the chitooligosaccharide peptide magnesium is 6.6, and the chitooligosaccharide peptide magnesium also comprises gibberellin and indoleacetic acid, wherein the mass ratio of the chitooligosaccharide peptide magnesium to the gibberellin to the indoleacetic acid is 30:35: 35.
The preparation method of the chitooligosaccharide peptide magnesium comprises the following steps:
performing enzymolysis on chitosan in acid environment with lysozyme at 50 deg.C for 24 hr to obtain agricultural chitosan oligosaccharide enzymolysis solution
And drying the agricultural chitosan oligosaccharide enzymolysis liquid to obtain the agricultural chitosan oligosaccharide for enzymolysis.
The acid environment is obtained by mixing distilled water, maleic anhydride and ammonium sulfate according to the mass ratio of 73:15:12, adding 5% of sodium hydroxide after mixing, and adjusting the pH value to 4.2;
the mass ratio of the chitosan, the acidic environment and the lysozyme is 3:96.9: 0.1.
Reacting for 40min under the conditions that the pressure is 10atm and the temperature is 160 ℃;
adding 10% sodium hydroxide water solution, and hydrolyzing to obtain agricultural chitooligosaccharide peptide hydrolysate.
The mass ratio of the 10 percent sodium hydroxide aqueous solution to the chitosan oligosaccharide enzymolysis solution is 21: 79; the hydrolysis time was 3 min;
adding magnesium nitrate into the agricultural oligo chitose peptide hydrolysate to obtain liquid oligo chitose peptide magnesium;
drying the liquid chitooligosaccharide peptide magnesium to obtain chitooligosaccharide peptide magnesium;
the molar ratio of magnesium ions to maleic acid in the magnesium nitrate was 0.5.
Gibberellin and indoleacetic acid are added into the chitooligosaccharide peptide magnesium to obtain a chitooligosaccharide peptide magnesium mixture.
The application method of the chitooligosaccharide peptide magnesium comprises the steps of diluting the chitooligosaccharide peptide magnesium by 1000 times with water to obtain a diluent, and spraying the diluent on the surface of a fruit 24 hours before heavy rain by adopting a spraying mode.
Diluting the chitooligosaccharide peptide magnesium mixture by water until the gibberellin content and the indoleacetic acid content in the diluent are both 100mg/L, and obtaining the chitooligosaccharide peptide magnesium diluent.
Example 5
A method for improving plant stress resistance and promoting flower bud differentiation comprises spraying chitooligosaccharide magnesium diluent on leaf surface 4 days after fruit picking in autumn.
The pH of the chitooligosaccharide peptide magnesium is 6.6, and the chitooligosaccharide peptide magnesium also comprises sodium indolylate and potassium indolybutyrate, wherein the mass ratio of the chitooligosaccharide peptide magnesium to the sodium indolylate to the potassium indolybutyrate to the potassium fulvate is 20:20:20: 40.
The preparation method of the chitooligosaccharide peptide magnesium comprises the following steps:
performing enzymolysis on chitosan in acid environment with lysozyme at 35 deg.C for 36 hr to obtain agricultural chitosan oligosaccharide enzymolysis solution
And drying the agricultural chitosan oligosaccharide enzymolysis liquid to obtain the agricultural chitosan oligosaccharide for enzymolysis.
The acid environment is obtained by mixing deionized water, maleic anhydride, glutaconic acid, monoammonium phosphate and ammonium sulfate according to the mass ratio of 74:5:8:3:10, adding 5% of sodium hydroxide after mixing, and adjusting the pH value to 5.0;
the mass ratio of the chitosan, the acidic environment and the lysozyme is 7:92.8: 0.2.
Reacting for 40min under the conditions that the pressure is 40atm and the temperature is 260 ℃;
adding 12% potassium hydroxide water solution, and hydrolyzing to obtain agricultural chitooligosaccharide peptide hydrolysate.
The mass ratio of the 12 percent sodium hydroxide aqueous solution to the chitosan oligosaccharide enzymolysis solution is 20: 80; the hydrolysis time was 3 min.
Adding magnesium nitrate into the agricultural oligo chitose peptide hydrolysate to obtain liquid oligo chitose peptide magnesium;
drying the liquid chitooligosaccharide peptide magnesium to obtain chitooligosaccharide peptide magnesium;
the molar ratio of magnesium ions to maleic acid in the magnesium nitrate was 0.5.
Adding sodium indoleacetate, potassium indolebutyrate and potassium fulvate into the chitooligosaccharide peptide magnesium to obtain the chitooligosaccharide peptide magnesium mixture.
Diluting chitooligosaccharide peptide magnesium with water 800 times to obtain chitooligosaccharide peptide magnesium diluent.
The application method of the chitooligosaccharide peptide magnesium comprises the steps of diluting a mixture of the chitooligosaccharide peptide magnesium by 800 times with water to obtain a diluent, and spraying the diluent on the surface of a fruit 24 hours before heavy rain by adopting a spraying mode.
Example 6
A method for improving plant stress resistance and promoting flower bud differentiation comprises spraying chitooligosaccharide magnesium diluent on leaf surface 4 days after fruit picking in autumn.
The pH of the chitooligosaccharide peptide magnesium is 6.6, and the chitooligosaccharide peptide magnesium also comprises sodium indolate and potassium indolate butyrate, wherein the mass ratio of the chitooligosaccharide peptide magnesium, the sodium indolate, the potassium indolate butyrate and the potassium fulvate is 20:15:30: 35.
The preparation method of the chitooligosaccharide peptide magnesium comprises the following steps:
performing enzymolysis on chitosan in acid environment with lysozyme at 50 deg.C for 24 hr to obtain agricultural chitosan oligosaccharide enzymolysis solution
And drying the agricultural chitosan oligosaccharide enzymolysis liquid to obtain the agricultural chitosan oligosaccharide for enzymolysis.
The acid environment is obtained by mixing distilled water, maleic anhydride and ammonium sulfate according to the mass ratio of 73:15:12, adding 5% of sodium hydroxide after mixing, and adjusting the pH value to 5.0;
the mass ratio of the chitosan, the acidic environment and the lysozyme is 3:96.9: 0.1.
Reacting for 40min under the conditions that the pressure is 10atm and the temperature is 160 ℃;
adding 10% sodium hydroxide water solution, and hydrolyzing to obtain agricultural chitooligosaccharide peptide hydrolysate; or
And drying the agricultural chitosan oligosaccharide hydrolysate to obtain the hydrolyzed agricultural chitosan oligosaccharide.
The mass ratio of the 10 percent sodium hydroxide aqueous solution to the chitosan oligosaccharide enzymolysis solution is 21: 79; the hydrolysis time was 3 min.
Adding magnesium chloride into the agricultural chitooligosaccharide peptide hydrolysate to obtain liquid chitooligosaccharide peptide magnesium;
drying the liquid chitooligosaccharide peptide magnesium to obtain chitooligosaccharide peptide magnesium;
the molar ratio of magnesium ions to maleic acid in the magnesium nitrate was 0.4.
Adding sodium indoleacetate, potassium indolebutyrate and potassium fulvate into the chitooligosaccharide peptide magnesium to obtain the chitooligosaccharide peptide magnesium mixture.
Diluting chitooligosaccharide peptide magnesium with water by 800 times to obtain chitooligosaccharide peptide magnesium diluent. Or the like, or, alternatively,
diluting the mixture of chitooligosaccharide peptide and magnesium by 800 times with water to obtain chitooligosaccharide peptide and magnesium diluent.
The beneficial effects of the present invention are further illustrated below in conjunction with experimental data:
experiment one
Test material
1, materials and methods:
1.1 test site: hipport fruit Biotech Co., Ltd.
1.2 test detection: content of chitosan oligosaccharide.
1.3 test materials: comparative example 1 (except that lysozyme was replaced by cellulase, the preparation method was the same as that in example 1), comparative example 2 (except that the preparation process of chitosan oligosaccharide produced by this company was the same as that in example 1 except that the preparation process was the same as that in example 3 in CN 106755208), comparative example 3 (except that tap water, the preparation method was the same as that in example 1), and the agricultural chitooligosaccharide enzymatic hydrolysate prepared in example 1.
1.4 Experimental implementation: the method comprises the steps of performing parallel experiments for three times respectively aiming at different batches of crude enzyme liquid and lysozyme of paecilomyces lilacinus provided by the same manufacturer, wherein the activities of the crude enzyme liquid and the lysozyme of the paecilomyces lilacinus are both 1000 wu/g, and detecting the content of chitosan oligosaccharide in each experiment.
The method is consistent with other management methods except different processing.
2 results and analysis
The content (%) of chitosan oligosaccharide is shown in Table 1.
TABLE 1
As can be seen from the comparison of the data of comparative examples 1 and 2 and example 1, the quality of comparative examples 1 and 2 is unstable, while the quality of comparative examples 3 and example 1 using lysozyme is stable.
As can be seen from the data of example 1 and comparative example 3, the content of chitosan oligosaccharide in example 1 using deionized water was high.
Experiment two
Test material
1, materials and methods:
1.1 test site: hipport fruit Biotech Co., Ltd.
1.2 test detection: the experimental phenomenon was observed.
1.3 test materials: 200ml of the agricultural chitosan oligosaccharide hydrolysate prepared in example 1 and 200ml of the agricultural chitosan oligosaccharide hydrolysate prepared in example 1 were prepared by dividing the agricultural chitosan oligosaccharide hydrolysate into two portions each of 100ml, adjusting the pH of the chitosan oligosaccharide hydrolysate to 6.5, and dividing the agricultural chitosan oligosaccharide hydrolysate into two portions each of 100ml, one of which was adjusted to 6.5, and the other was adjusted to 4.5.
1.4 Experimental implementation: preparing a 5% magnesium nitrate solution, adding magnesium nitrate into the two chitosan oligosaccharide enzymolysis solutions and 1 part of agricultural chitosan oligosaccharide hydrolysis solution respectively, wherein the adding amount of the magnesium nitrate is that the molar ratio of magnesium ions to maleic acid in the final solution is 0.8, adding magnesium nitrate into the other agricultural chitosan oligosaccharide hydrolysis solution, the adding amount of the magnesium nitrate is calculated by magnesium ions, and the molar ratio of the magnesium ions to maleic acid in the agricultural chitosan oligosaccharide hydrolysis solution is 1.1.
The method is consistent with other management methods except different processing.
2 phenomena of experiment
Adding magnesium nitrate into agricultural chitosan oligosaccharide enzymolysis liquid with pH of 6.5 to generate a large amount of white flocculation, standing for 1h, and flocculating and settling at the bottom.
Adding magnesium nitrate into agricultural chitooligosaccharide enzymolysis liquid with pH of 4.5, standing for 1 hr without white flocculation, and no precipitate at the bottom.
Adding magnesium nitrate, magnesium ions and maleic acid with a molar ratio of 0.8 into the mixture, wherein the chitosan oligosaccharide hydrolysate does not generate flocculation, and standing for 1h, and no precipitate is generated at the bottom.
Adding magnesium nitrate, wherein the molar ratio of the added magnesium ions to the maleic acid in the agricultural chitosan oligosaccharide hydrolysate is 1.1, and precipitates are formed at the bottom.
Therefore, the chitosan oligosaccharide peptide magnesium prepared by the method has good stability, and can solve the problem that chitosan oligosaccharide and magnesium do not coexist in an environment with neutral pH.
Experiment three
Test material
1, materials and methods:
1.1 test site: the Shandong tobacco Taixi Yuan has a tree age of 9 years and the apple variety is Fuji.
1.2 test detection: lignin, pentosan, defoliation time, number of results, grade and total apple weight, wherein defoliation time is based on the number of leaves remaining below 50.
1.3 test materials: blank (15-15-15 compound fertilizer), comparative 4 (gibberellin), comparative 5 (same as example 2 except that it is sprayed on day 12 after apple picking), comparative 6 (magnesium nitrate is added to the chitooligosaccharide enzymatic hydrolysate prepared in example 1, pH is 4.5, and magnesium is added in the same amount as example 2), liquid magnesium chitooligosaccharide peptide prepared in example 1 and liquid magnesium chitooligosaccharide peptide mixture prepared in example 2.
1.4 Experimental implementation: the liquid chitooligosaccharide peptide magnesium mixture prepared in comparative example 4 (gibberellin) and example 2 was diluted with water to have a gibberellin content of 100mg/L, the liquid chitooligosaccharide peptide magnesium prepared in example 1 was diluted with water to have a chitosan oligosaccharide content consistent with that of example 2 after dilution, blank (compound fertilizer of 15-15-15) bases were applied to the bottom of a tree, 1.5kg per fruit base, comparative example 5 (the operation was consistent with example 2 except for spraying on the 12 th day after picking apples) was diluted to have a chitosan oligosaccharide content consistent with example 2, and comparative example 6 (prepared by adding magnesium nitrate to the chitooligosaccharide enzymatic hydrolysate prepared in example 1, pH was 4.5, and the prepared chitooligosaccharide magnesium was diluted with water to have a chitosan oligosaccharide content consistent with that of example 1).
Taking 50L of each of the diluents (except for blank and comparison 5), spraying leaf surfaces of fruit trees at 4 nightfall, wherein the number of treated apple trees is 100, 500ml of the treatment liquid is sprayed on each apple tree, the treatment time is 2019, 10 and 19 days, the apple picking time is 2019, 10 and 16 days, blank (15-15-15 compound fertilizer) is applied to the bottoms of the fruit trees in a ditch of 2019, 10 and 19 days, 1.5kg of the base of each fruit tree is applied, and the number of treated apple trees is 100; and (3) preparing 50L in comparison with 5, spraying the leaf surfaces of the fruit trees at the evening 4, wherein the number of treated apple trees is 100, 500ml of treatment liquid is sprayed on each apple tree, and the treatment time is 2019, 10 months and 28 days.
Counting fruits in the second year, recording the number of fruits, reserving 135 fruits per apple tree, and evaluating the apple grade after harvesting in autumn.
The method is consistent with other management methods except different processing.
Grade evaluation: 80% of apples have a diameter of more than 80cm, preferably; 60-80% of apples have the diameter of more than 80cm and are good; 40-60% of apples have the diameter larger than 80cm, which is common; less than 40% of the apples were greater than 80cm in diameter, which is a difference.
2 results and analysis
Lignin, pentosan, defoliation time (2019, only month and day), number of results, grade and total weight (kg) are shown in Table 2
TABLE 2
| Lignin (%) | Pentosan (%) | Fallen leaves time (moon day) | Fruit number (pieces) | Grade | Total weight (kg) | |
| Blank space | 22.21 | 23.75 | 11.15 | 1896 | Good wine | 3459.5 |
| Comparative example 4 | 23.58 | 24.86 | 11.20 | 2069 | Superior food | 3562.3 |
| Comparative example 5 | 22.08 | 23.16 | 11.14 | 1872 | Good wine | 3467.2 |
| Comparative example 6 | 22.96 | 24.31 | 11.17 | 1995 | Good wine | 3519.6 |
| Example 1 | 23.53 | 24.82 | 11.19 | 2078 | Superior food | 3546.7 |
| Example 2 | 24.15 | 25.27 | 11.24 | 2162 | Superior food | 3604.5 |
As can be seen from the comparison between the blank and the data in example 1 in table 2, the content of lignin and pentosan in the tree can be significantly increased, the content of lignin and pentosan is increased, the freezing injury can be effectively resisted, and the stress resistance of the fruit tree is improved; as can be seen from comparison of the data of comparative example 6 and example 1, although chitosan oligosaccharide and magnesium ion can coexist in an environment of pH 4.5, they are not as effective as example 1 in increasing the lignin and pentosan content and are also not as effective as example 1 in preventing defoliation; as can be seen from comparison of data in comparison 5 and example 2, the spraying time directly affects the use effect, and the spraying time in example 2 immediately after picking can better improve the contents of lignin and pentosan in the tree body, improve the stress resistance of the tree body, and meanwhile, can enable the leaf surface to be stored on the tree for a longer time, so as to better supplement the nutrition of the tree body; it can be seen from the data of example 1, comparison 4 and example 2 that the magnesium chitooligosaccharide peptide and the auxin can mutually promote, the auxin provides guarantee for the absorption of the chitooligosaccharide, the chitooligosaccharide provides nutrition for the growing leaves, so that the leaves can not senilis due to the nutrition consumption in the growth, the two mutually presuppose, the stress resistance of the tree body can be better improved, the flower bud differentiation can be promoted, and the quality and the yield of the fruit can be finally improved.
Claims (7)
1. A method for improving the stress resistance of plants and promoting flower bud differentiation is characterized in that within 10 days after fruits are picked in autumn, a chitosan oligosaccharide peptide magnesium diluent is sprayed on leaf surfaces;
the chitooligosaccharide peptide magnesium diluent is prepared according to the following steps:
carrying out enzymolysis on chitosan in an acid environment by lysozyme to obtain agricultural chitosan oligosaccharide enzymolysis liquid;
heating the chitosan oligosaccharide enzymolysis liquid under certain conditions;
adding alkali for hydrolysis to obtain chitosan oligosaccharide peptide hydrolysate;
adding magnesium ions to obtain liquid chitooligosaccharide magnesium; or the like, or, alternatively,
drying the liquid chitooligosaccharide peptide magnesium to obtain chitooligosaccharide peptide magnesium;
diluting the liquid chitooligosaccharide peptide magnesium to obtain a diluent; or
Diluting the chitooligosaccharide peptide magnesium to obtain a diluent;
the chitooligosaccharide peptide magnesium is diluted by water, and the EC value of the diluted solution is lower than 4 mS/cm;
the certain conditions are that the pressure is 3 atm-45 atm, the heating temperature is 130-280 ℃, and the heating time is 20-60 min;
the mass ratio of the alkali to the chitosan oligosaccharide enzymolysis liquid is 2-40: 60-98; the hydrolysis time is 2-30 min;
the alkali is one or more of sodium hydroxide, potassium hydroxide and their water solution;
the acid environment comprises water, water-soluble unsaturated dibasic organic acid or water-soluble unsaturated dibasic organic acid anhydride and ammonium ions;
the pH value of the acidic environment is 4.0-5.5;
the enzymolysis is carried out for 6-48 h at the temperature of 30-60 ℃;
the mass ratio of the chitosan to the acid environment to the lysozyme is 1-10: 89.9-98.9: 0.1-1;
the mass ratio of water, water-soluble unsaturated dibasic organic acid or water-soluble unsaturated dibasic organic anhydride and ammonium ions in the acidic environment is 60-92: 6.5-39: 1-8;
the molar ratio of the added magnesium ions to the water-soluble unsaturated dibasic organic acid or the water-soluble unsaturated dibasic organic acid anhydride in the agricultural chitooligosaccharide peptide hydrolysate is less than 1.
2. The method for increasing the stress tolerance and promoting flower bud differentiation of a plant according to claim 1, wherein said chitooligosaccharide peptide comprises one or both of auxin and cytokinin.
3. The method for improving plant stress resistance and promoting flower bud differentiation according to claim 1 or 2, wherein the pH of the chitooligosaccharide peptide magnesium is 6.0-7.0.
4. The method for improving stress resistance of plants and promoting flower bud differentiation according to claim 2,
mixing one or two of auxin and cytokinin with liquid chitooligosaccharide peptide magnesium or chitooligosaccharide peptide magnesium to obtain chitooligosaccharide peptide magnesium mixture.
5. The method for improving the stress resistance and promoting flower bud differentiation of a plant according to claim 4, wherein said blending comprises dissolving and mixing.
6. The method for improving plant stress resistance and promoting flower bud differentiation according to claim 4 or 5,
diluting the chitooligosaccharide peptide magnesium mixture to obtain chitooligosaccharide peptide magnesium diluent.
7. The method for improving stress resistance and promoting flower bud differentiation of a plant according to claim 6,
the chitooligosaccharide peptide magnesium mixture is diluted by water until the content of auxin or cytokinin in the diluted solution is 100 mg/L-150 mg/L, and the EC value of the diluted solution is lower than 4 mS/cm.
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Denomination of invention: A method for improving plant stress resistance and promoting flower bud differentiation Effective date of registration: 20230703 Granted publication date: 20220329 Pledgee: Anqiu Shandong rural commercial bank Limited by Share Ltd. Pledgor: Shandong Aiguozhi Biotechnology Co.,Ltd. Registration number: Y2023980046565 |