CN116831125A - Oligosaccharide biological stimulant and application method thereof - Google Patents

Oligosaccharide biological stimulant and application method thereof Download PDF

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Publication number
CN116831125A
CN116831125A CN202310797215.XA CN202310797215A CN116831125A CN 116831125 A CN116831125 A CN 116831125A CN 202310797215 A CN202310797215 A CN 202310797215A CN 116831125 A CN116831125 A CN 116831125A
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oligosaccharide
component
biostimulant
mixing
crops
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刘海玉
王祥传
成道泉
王殿海
王程璐
陈晓洁
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Shandong Sgotts Plant Nutrition Technology Co ltd
Shandong Jingbo Agrochemical Technology Co ltd
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Shandong Sgotts Plant Nutrition Technology Co ltd
Shandong Jingbo Agrochemical Technology Co ltd
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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
    • A01N43/14—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 six-membered rings
    • A01N43/16—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 six-membered rings with oxygen as the ring hetero atom
    • 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
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P21/00—Plant growth regulators

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  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Environmental Sciences (AREA)
  • Botany (AREA)
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  • Pest Control & Pesticides (AREA)
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  • Chemical & Material Sciences (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Dentistry (AREA)
  • Agronomy & Crop Science (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)

Abstract

The invention belongs to the technical field of biostimulants, and particularly relates to an oligosaccharide biostimulant, wherein the active ingredients of the biostimulant comprise a component A, a component B and a component C; the biological stimulant can improve the activity of crops and increase the yield and quality of agricultural products, and simultaneously solves the problems of growth inhibition, overgrowth or weak immunity caused by using a single oligosaccharide.

Description

Oligosaccharide biological stimulant and application method thereof
Technical Field
The invention belongs to the technical field of biostimulants, and particularly relates to an oligosaccharide biostimulant and a use method thereof.
Background
For many years, abiotic stresses such as drought, waterlogging, low temperature, high temperature, insufficient illumination and the like and biotic stresses such as diseases, insect pests, grass pests and the like have great negative effects on the yield and quality of agricultural products. The use of fertilizers and chemical pesticides can alleviate the loss caused by the stress to a certain extent, but the unreasonable use of fertilizers and pesticides throughout the year brings hidden danger to the environment and human safety. Therefore, the use of the safer, more effective biological source active substance, biostimulant (biostimulnt), is of interest.
Biostimulants, also known as biostimulants, plant vigor agents, refer to a substance comprising certain components and microorganisms whose efficacy, when applied to a plant or root canal, is stimulating to the natural processes of the plant, including enhancing/contributing to nutrient absorption, nutrient efficacy, abiotic stress resistance and crop quality, independently of the nutrient components. Although the biostimulant is not a plant hormone or a nutrient component, the biostimulant stimulates the vigor of plants in a very small amount to promote the growth and development.
Chitosan oligosaccharides are generally obtained from shrimp shell, crab shell or fungal mycelium as raw materials by a series of physical, chemical and/or biological treatments, both of which are exogenous biostimulants for plants. Pectin oligosaccharide, cellooligosaccharide and xylooligosaccharide are derived from plants and belong to endogenous biological hormone. Both types of biostimulants can activate the immune response of plant cells, but the receptors on cell membranes are different, and the influence on the physiological and biochemical processes of the plant cells is also greatly different. Alginate-oligosaccharide (ADO) is a small molecular weight fragment obtained from Alginate in brown algae by oxidative degradation, acid hydrolysis or lyase degradation. ADO can promote the production of plant endogenous hormones, increase the net photosynthetic rate, and further promote crop growth.
At present, scientists have found receptors for chitosan oligosaccharide on cell membranes of various crops such as arabidopsis thaliana, rice and the like, and have elaborated the mechanism of action of the receptor. The oligogalacturonate may interact with a cell wall-linked receptor-like kinase comprising an extracellular domain, a transmembrane domain and an intracellular domain, wherein the extracellular domain is directly linked to a cell wall component and the intracellular domain has serine/threonine protein kinase activity and thus has the ability to transmit signals across the membrane. The glucan oligosaccharides (including cellooligosaccharides) are the first oligosaccharide inducers found and thus the first oligosaccharide inducers to be studied in the acceptor. As early as 1983, scientists labeled branched beta-1, 3-glucan extracted from phytophthora sojae with isotopes, found that it specifically bound to soybean She Cunzai; after that, it was found that the dextran specifically binds to soybean suspension cells, protoplasts and root-derived cells, and that the binding mainly occurs on plasma membranes.
Multiple signal paths exist in plant immunity, the recognition capability of different crops on different oligosaccharide biological stimulators is different, and meanwhile, a certain effect offset or superposition amplification effect can exist between different immunity signal paths. Therefore, the influence of different oligosaccharide compositions on the physiology, growth and the like of crops is studied, the knowledge of the products can be deepened from the application level, and a new product with general immunocompetence and more outstanding efficacy on the crops can be developed with a larger opportunity.
Disclosure of Invention
In order to solve the problems, the invention provides an oligosaccharide biological stimulator and a using method thereof, and the biological stimulator can improve the activity of crops and the yield and quality of agricultural products, and simultaneously solve the problems of growth inhibition, overgrowth or weak immunity caused by using a single oligosaccharide.
The inventor carries out intensive research to solve the existing problems, and repeatedly carries out experiments, and the result shows that when the mixture consisting of the chitosan oligosaccharide and the brown alginate oligosaccharide is applied with any one or more of pectin oligosaccharide, fiber oligosaccharide and xylooligosaccharide, the activity of single oligosaccharide can be greatly improved, the growth of crops can be promoted, and the yield and quality of agricultural products can be improved.
The technical scheme of the invention is as follows:
an oligosaccharide biological stimulant comprises an active ingredient of a component A, a component B and a component C; the component A is chitosan oligosaccharide, the component B is brown alginate oligosaccharide, and the component C is one or more of pectin oligosaccharide, cellooligosaccharide and xylooligosaccharide in any proportion.
Preferably, the stimulator comprises 1-5wt% of component A, 1-5wt% of component B, 1-5wt% of component C, 1.1-6wt% of auxiliary materials, and the balance of solvent, wherein the pH value is 4.0-8.0.
Preferably, the degree of polymerization of the active ingredients is in the range of 2 to 10. Further preferably 4-10.
The application method of the oligosaccharide biological stimulant comprises spraying or applying the oligosaccharide biological stimulant on the leaves or roots of crops. The biostimulant and the fertilizer can be sprayed or applied to the roots of the crop simultaneously during application.
Preferably, the concentration of the active ingredient of the biostimulant for crops is 0.5-200ppm. Further preferably, the biostimulant is used for crops, the concentration of active ingredient for foliar application is 20-200ppm, and the concentration of active ingredient for root application is 10-120ppm.
Preferably, the crop is one of Compositae, solanaceae, brassicaceae, gramineae, leguminosae, rosaceae, cucurbitaceae, convolvulaceae, chenopodiaceae, liliaceae, umbelliferae, malvaceae, zingiberaceae, and Nelumbo nucifera.
Preferably, the auxiliary materials comprise a surfactant and a preservative, and the solvent is water. It is further preferred that the surfactant is 1-5wt% and the preservative is 0.1-1.0wt% in terms of weight percent, wherein the surfactant is betaine and the preservative is sodium diacetate.
Specifically, the crop may be plants of Brassicaceae such as cabbage, green cauliflower, radish, turnip, etc., plants of Solanaceae such as potato, tomato, eggplant, bell pepper, capsicum, pepper, tobacco, etc., plants of Asteraceae such as crowndaisy, lettuce, burdock, etc., plants of Cucurbitaceae such as watermelon, melon, pumpkin, cucumber, bitter gourd, towel gourd, etc., plants of Chenopodiaceae such as spinach, beet, plants of Umbelliferae such as carrot, celery, parsley, and Cryptotaenia, plants of Leguminosae such as soybean, red bean, broad bean, pea, tetrapanax, peanut, etc., plants of Brassicaceae such as sweet potato, plants of Liliaceae such as leek, shallot, onion, garlic, etc., plants of Rosaceae such as strawberry, apple, pear, loquat, etc., plants of Malvaceae such as okra, etc., plants of Zingiberaceae such as ginger, lotus, etc., plants of Gramineae, corn, rice, barley, wheat, sugarcane, etc.
In the invention, the chitosan oligosaccharide is obtained by degrading chitin, wherein the chitin is polysaccharide formed by connecting beta-N-acetyl-D-glucosamine with beta-1, 4 glycosidic bond, is second largest natural polysaccharide with content except cellulose in nature, widely exists in shrimp, crab shells, insect exoskeletons and fungus cell walls, and has higher immune induction resistance activity than the chitosan oligosaccharide with the polymerization degree of 2-3, namely the chitosan oligosaccharide with the polymerization degree of 4-8.
The chitosan oligosaccharide used in the present invention is particularly preferably a substance having the following chemical structure.
In addition, acetyl groups (COCH) in the formula are also included 3 ) Partially fall off and become NH 2 The ratio of such deacetylation is preferably 30% or less of the total chitosan oligosaccharide.
The application of the stimulant of the invention can produce plants or parts thereof (such as roots, stems, leaves, flowers, fruits, seeds, tissues, cells and the like) with the activity of a plant vaccine, so that the vigor of crops can be improved, the resistance of the crops to biotic stress and abiotic stress can be improved, and the harvest quantity, the quality and the shelf life after harvest are also obviously improved. The biological stimulant can strongly excite the resistance of crops to biological stress, and can keep the crops in a good state under multiple adverse conditions, thereby laying a good foundation for stabilizing/improving the yield and the quality.
The activity of biostimulants is determined primarily by measuring chitinase production in plants, while other indicators monitored include: root system and overground biomass, root system activity, chlorophyll content, endogenous hormone level, etc. The chitinase production is used as one of the characterization objects of the effect of the biostimulant, and the chitinase production reflects the defensive capability of crops when the crops are subjected to external pressure, and part of leaves of plants in cultivation are adopted to analyze the chitinase activity of the plants so as to evaluate different treatments. According to the invention, by measuring indexes such as chitinase activity, root system and upper biomass, root system activity, chlorophyll content, endogenous hormone level and the like, the biological stimulator provided by the invention has obvious growth promoting and immune activating capabilities, effectively balances the relationship between crop growth and immunity, promotes crop growth without sacrificing disease resistance and improves the quality and yield of products.
The oligosaccharide biological stimulator and the application method thereof provided by the invention have the advantages that the active ingredients of the biological stimulator are composed of different oligosaccharides, the functions of improving stress resistance of crops, promoting growth of the crops and improving quality and yield are realized, the vitality of the crops can be improved, the yield and quality of agricultural products can be improved, and the problems of growth inhibition, overgrowth or weak immunity caused by using a single oligosaccharide are avoided.
Drawings
FIG. 1 shows a standard curve of chitinase content.
Detailed Description
The technical scheme of the invention is described below through specific examples. The raw materials and reagents used in the present invention are commercially available, and the following percentages are weight percentages.
The specific technical indexes are as follows:
the single oligosaccharide or the composition can influence physiological metabolism activities in the cells of crops, such as inducing chitinase to generate, promoting synthesis of chlorophyll and endogenous hormone, and the like after being applied to the crops. The chitinase content in the leaves was determined using a kit (Solarbio, beijing, china). Chitinase Activity definition: the amount of enzyme that produces 1 mu g N-acetamido glucose per gram of tissue per hour at 37℃is one unit of enzyme activity. The method mainly comprises the following steps:
the preparation method of the colloidal chitin comprises the following steps: weighing 20g of powdery chitin, slowly pouring into 200mL of concentrated hydrochloric acid, adding 200mL of distilled water, continuously stirring for 12h by using a magnetic stirrer, adding 2L of 95% ethanol at 4 ℃, standing for 12h at room temperature after uniformly mixing, and centrifuging for 20min at 4 ℃ at 5000 r/min. Repeatedly washing the precipitate with distilled water to neutrality, and finally keeping the volume to 1L with distilled water, and storing at 4deg.C for use.
(II) preparation of chitinase content standard curve: a standard curve was established based on the concentration (x, μg/mL) of the N-acetylglucosamine standard tube and the absorbance ΔA standard (y, ΔA standard).
(III) preparation of leaf extract, adding 1.0mL of protein extraction buffer into a 2.0mL centrifuge tube, adding 0.10g (marked as W) of leaf cut into about 3X 3mm by scissors, processing each sample for 3 times, further mashing the leaf with a glass rod until no macroscopic solid exists, centrifuging at 10000r/min and 4 ℃ for 20 minutes, recovering a water layer into a new 2.0mL centrifuge tube, preparing an extract, and placing the extract into ice water for test.
(IV) substituting delta A into an equation according to a standard curve to obtain the content x (mug/mL) of chitinase in the sample; the enzyme activity (U/mg) =x/W/1000 was calculated from the sample mass.
Example 1
An oligosaccharide biological stimulant: 1.7% of chitosan oligosaccharide, 1.7% of brown alginate oligosaccharide, 1.7% of cellooligosaccharide, 1.0% of betaine, 0.3% of sodium diacetate and the balance of water;
the specific preparation steps of the solution are as follows: mixing chitosan oligosaccharide, brown alginate oligosaccharide, cellooligosaccharide, sodium diacetate and water according to the above formula, shearing and mixing at high speed, adding betaine, continuously shearing and mixing, and adjusting pH to 7.0 to obtain the final product.
Example 2
An oligosaccharide biological stimulant: 1.7% of chitosan oligosaccharide, 1.7% of brown alginate oligosaccharide, 1.7% of pectin oligosaccharide, 1.0% of betaine, 0.3% of sodium diacetate and the balance of water;
the specific preparation steps of the solution are as follows: mixing chitosan oligosaccharide, brown alginate oligosaccharide, pectin oligosaccharide, sodium diacetate and water according to the above formula, shearing and mixing at high speed, adding betaine, continuously shearing and mixing, and adjusting pH to 7.0 to obtain the final product.
Example 3
An oligosaccharide biological stimulant: 1.7% of chitosan oligosaccharide, 1.7% of brown alginate oligosaccharide, 1.7% of xylooligosaccharide, 1.0% of betaine, 0.3% of sodium diacetate and the balance of water;
the specific preparation steps of the solution are as follows: mixing chitosan oligosaccharide, brown alginate oligosaccharide, xylooligosaccharide, sodium diacetate and water according to the above formula, shearing and mixing at high speed, adding betaine, continuously shearing and mixing, and obtaining pH 7.0.
Example 4
An oligosaccharide biological stimulant: 1% of chitosan oligosaccharide, 1% of brown alginate oligosaccharide, 2% of cellooligosaccharide, 1.0% of betaine, 0.3% of sodium diacetate and the balance of water;
the specific preparation steps of the solution are as follows: mixing chitosan oligosaccharide, brown alginate oligosaccharide, cellooligosaccharide, sodium diacetate and water according to the above formula, shearing and mixing at high speed, adding betaine, continuously shearing and mixing, and adjusting pH to 7.0 to obtain the final product.
Example 5
An oligosaccharide biological stimulant: 2% of chitosan oligosaccharide, 4% of brown alginate oligosaccharide, 1% of pectin oligosaccharide, 1.0% of betaine, 0.3% of sodium diacetate and the balance of water;
the specific preparation steps of the solution are as follows: mixing chitosan oligosaccharide, brown alginate oligosaccharide, pectin oligosaccharide, sodium diacetate and water according to the above formula, shearing and mixing at high speed, adding betaine, continuously shearing and mixing, and adjusting pH to 7.0 to obtain the final product.
Example 6
An oligosaccharide biological stimulant: 5% of chitosan oligosaccharide, 1% of brown alginate oligosaccharide, 3% of xylooligosaccharide, 1.0% of betaine, 0.3% of sodium diacetate and the balance of water;
the specific preparation steps of the solution are as follows: mixing chitosan oligosaccharide, brown alginate oligosaccharide, xylooligosaccharide, sodium diacetate and water according to the above formula, shearing and mixing at high speed, adding betaine, continuously shearing and mixing, and obtaining pH 7.0.
Comparative example 1
An oligosaccharide biological stimulant: 5.0% of chitosan oligosaccharide, 1.0% of betaine, 0.3% of sodium diacetate and the balance of water.
The specific preparation steps of the solution are as follows: mixing chitosan oligosaccharide, sodium diacetate and water according to the formula, uniformly shearing and mixing at high speed, adding betaine, continuously shearing and uniformly mixing, and obtaining the pH value of 7.0, thereby completing the preparation of the solution.
Comparative example 2
An oligosaccharide biological stimulant: brown alginate oligosaccharide 5.0%, betaine 1.0%, sodium diacetate 0.3% and water balance.
The specific preparation steps of the solution are as follows: mixing brown alginate oligosaccharides, sodium diacetate and water according to the above formula, shearing and mixing at high speed, adding betaine, continuously shearing and mixing, and obtaining pH 7.2 to obtain the final product.
Comparative example 3
An oligosaccharide biological stimulant is prepared from cellooligosaccharide 5.0%, betaine 1.0%, sodium diacetate 0.3%, and water in balance.
The specific preparation steps of the solution are as follows: mixing cellooligosaccharide, sodium diacetate and water according to the formula, uniformly shearing and mixing at a high speed, adding betaine, continuously shearing and uniformly mixing, and obtaining the pH value of 7.1 to finish the preparation of the solution.
Comparative example 4
An oligosaccharide biological stimulant: pectin oligosaccharide 5.0%, betaine 1.0%, sodium diacetate 0.3%, and water balance.
The specific preparation steps of the solution are as follows: mixing pectin oligosaccharide, sodium diacetate and water according to the above formula, shearing and mixing at high speed, adding betaine, continuously shearing and mixing, and obtaining the final product with pH of 6.8.
Comparative example 5
An oligosaccharide biological stimulant: xylo-oligosaccharide 5.0%, betaine 1.0%, sodium diacetate 0.3% and water balance.
The specific preparation steps of the solution are as follows: mixing xylooligosaccharide, sodium diacetate and water according to the formula, uniformly shearing and mixing at a high speed, adding betaine, continuously shearing and uniformly mixing, and obtaining the pH value of 6.6, thereby completing the preparation of the solution.
Comparative example 6
An oligosaccharide biological stimulant: 2.5% of chitosan oligosaccharide, 2.5% of brown alginate oligosaccharide, 1.0% of betaine, 0.3% of sodium diacetate and the balance of water.
The specific preparation steps of the solution are as follows: mixing chitosan oligosaccharide, brown alginate oligosaccharide, sodium diacetate and water according to the above formula, shearing and mixing at high speed, adding betaine, continuously shearing and mixing until the pH is 7.0, and completing the preparation of the solution.
Comparative example 7
An oligosaccharide biological stimulant: 2.5% of chitosan oligosaccharide, 2.5% of cellooligosaccharide, 1.0% of betaine, 0.3% of sodium diacetate and the balance of water.
The specific preparation steps of the solution are as follows: mixing chitosan oligosaccharide, cellooligosaccharide, sodium diacetate and water according to the above formula, shearing and mixing at high speed, adding betaine, continuously shearing and mixing to obtain pH 7.0, and preparing solution.
Comparative example 8
An oligosaccharide biological stimulant: 2.5% of chitosan oligosaccharide, 2.5% of pectin oligosaccharide, 1.0% of betaine, 0.3% of sodium diacetate and the balance of water.
The specific preparation steps of the solution are as follows: mixing chitosan oligosaccharide, pectin oligosaccharide, sodium diacetate and water according to the above formula, shearing and mixing at high speed, adding betaine, continuously shearing and mixing to obtain pH 7.0, and preparing solution.
Comparative example 9
An oligosaccharide biological stimulant: 2.5% of chitosan oligosaccharide, 2.5% of xylooligosaccharide, 1.0% of betaine, 0.3% of sodium diacetate and the balance of water.
The specific preparation steps of the solution are as follows: mixing chitosan oligosaccharide, xylooligosaccharide, sodium diacetate and water according to the formula, uniformly shearing and mixing at high speed, adding betaine, continuously shearing and uniformly mixing, and obtaining the pH value of 7.0 to finish the preparation of the solution.
Test example 1: promoting growth of root system and overground part of tomato
The test was performed in 2023 in glass greenhouse from Shandong Beijing Bonong chemical technology Co., ltd. The turfy soil (purchased in flower market) and sand are uniformly mixed according to the ratio of 1:1 and are filled into a 54cm multiplied by 27cm multiplied by 6cm seedling raising tray, the seeds are uniformly sown into the seedling raising tray and covered with a soil layer with the thickness of about 0.5cm, watering is carried out from the bottom, and the seeds are germinated and raised under the condition of abundant sunlight at 24-28 ℃. After tomato plants grow to 6-8 leaves, selecting seedlings with consistent growth vigor, transplanting the seedlings into flowerpots (the diameter of each flowerpot is 20cm, the height of each flowerpot is 15cm, each pot is filled with field soil for 4.0 kg), transplanting one seedling into each flowerpot, keeping the growth conditions of each pot seedling consistent, and applying a large amount of element water-soluble fertilizer (N-P) after seedling recovery 2 O 5 -K 2 O: 17-17-17), the water-soluble fertilizer is used according to the dosage of 30kg/hm 2 The biological stimulant and the macroelement water-soluble fertilizer are applied together, namely the products prepared in the examples and the comparative examples are firstly dispersed in the macroelement water-soluble fertilizer solution, the active ingredients of the products are diluted to be shown in the following table, the application amount of each plant is 100mL, and the clear water control group only applies the element water-soluble fertilizer solution with the same amount and concentration. The experiment was set up with 13 treatments, 18 seedlings per treatment, 6 seedlings in one group, three replicates. The experimental design is shown in table 1.
Table 1 test design
20 days after application of the above samples, 6 plants were randomly selected for each treatment group, root soil was carefully washed away, and the root dry weight (80 ℃ C. Treatment for 6 hours after draining) and the fresh weight on the ground were measured. 3 plants were additionally taken for each treatment, the root system was washed in an ice-water bath and 0.5g of fresh root tip was taken and the root system activity of the different treatment groups was determined by TTC staining. 3 plants were taken for each treatment and the chitinase activity, chlorophyll content (chlorophyll meter) and indoleacetic acid (see DB 22/T25989-2016) levels in leaf cells were determined. The specific results are shown in Table 2.
TABLE 2 comparison of tomato growth and physiological index for different treatment groups
According to the data in Table 2, chitooligosaccharides are capable of efficiently activating chitinase expression in crop cells, thereby improving the resistance of crops to external stress, but the acquisition of such ability affects biomass accumulation to some extent. The brown alginate oligosaccharides have excellent rooting and growth promoting capacity, but have low immune induction activity. The chitosan oligosaccharide and brown alginate oligosaccharide treatment group has higher capability of coping with external stress while promoting the growth of tomatoes. When cellooligosaccharide, pectin oligosaccharide and xylooligosaccharide are singly applied, the application of the cellooligosaccharide, pectin oligosaccharide and xylooligosaccharide is not similar to that of chitosan oligosaccharide, but the cellooligosaccharide, pectin oligosaccharide and xylooligosaccharide are combined and then applied, so that the synergistic effect of both the growth promotion and immune induction activities is achieved. Especially, the chitosan oligosaccharide, brown alginate oligosaccharide and cellooligosaccharide combined treatment group has the most obvious capability of promoting growth and activating immunity.
Test example 2: different treatments improve the disease resistance of plants to cucumber botrytis cinerea
A treatment group (Table 2) with higher growth promoting and chitinase activities was selected and further examined for its ability to prevent cucumber gray mold damage.
TABLE 3 experimental design
The method comprises the following specific steps: (1) prophylactic administration. The samples prepared in examples and comparative examples were diluted with clear water at a constant ratio to obtain the concentrations of the medicinal liquids shown in Table 3. Make the following stepsSpraying crops in each block by using a backpack sprayer, using equal amount of clear water in a clear water control group, continuously applying the pesticide twice, and investigating the disease condition after 5d after 1 st pesticide application and 10d after 2 nd pesticide application respectively at intervals of 7 days. The number of cucumber leaf disease stages was recorded according to the GB/T17980.28-2000 method. (2) Botrytis cinerea infects cucumber leaves. Transferring the separated Botrytis cinerea to PDA culture medium for activating culture (28 deg.C for 3 d), inoculating to PDB culture medium with 10mm puncher, shaking culture at 28 deg.C and 180rpm for 3d, filtering the culture solution with sterilized cotton, removing mycelium to obtain Botrytis cinerea spore suspension, measuring spore suspension concentration by blood cell plate counting method, and regulating spore number to 10 with sterile water 8 CFU/mL, after 2d of preventive administration, the spore suspension was diluted 500-fold and sprayed evenly onto cucumber leaves to cause disease.
TABLE 4 comparison of disease index for different treatment groups
The data in Table 4 show that the disease development can be effectively inhibited by using the binary composition composed of the chitosan oligosaccharide or the chitosan oligosaccharide and other four oligosaccharides, the disease development can be well inhibited by using the chitosan oligosaccharide and the brown alginate oligosaccharide to be matched with the cellooligosaccharide or the pectin oligosaccharide or the xylooligosaccharide, and the chitosan oligosaccharide, the brown alginate oligosaccharide and the cellooligosaccharide treatment group has the optimal effect in preventing the gray mold of the cucumber. In combination with the data in Table 3, the treatment group also has outstanding rooting and growth promoting effects, and the treatment group can effectively balance the relationship between crop growth and immunity, and does not sacrifice disease resistance while promoting crop growth.
Test example 3: oligosaccharide biostimulant for improving tomato quality
(1) Experimental sample and usage amount
The active ingredient concentrations in each sample and the use samples are shown in Table 5. Experimental groups the diluted solutions of the products of examples and comparative examples were applied on the basis of the control group, diluted to the concentrations of oligosaccharides (active ingredients) as shown in the following table, and the application of fertilizers and pesticides was the same as the conventional management. Root Shi Xishi solution was applied 3 times in succession at 20-day intervals from 10 days after tomato seedling transplanting and seedling reviving.
Table 5 samples and usage amounts
Treatment group Dosage of usage
Control group Conventional management
Chitosan oligosaccharide (comparative example 1) 100ppm, root irrigation, 100 mL/plant
Chitosan oligosaccharide and brown alginate oligosaccharide (comparative example 6) 50+50ppm, root irrigation, 100 mL/plant
Chitosan oligosaccharide and cellooligosaccharide (comparative example 7) 50+50ppm, root irrigation, 100 mL/plant
Chitosan oligosaccharide and pectin oligosaccharide (comparative example 8) 50+50ppm, root irrigation, 100 mL/plant
Chitosan oligosaccharide and xylooligosaccharide (comparative example 9) 50+50ppm, root irrigation, 100 mL/plant
Chitosan oligosaccharide, brown alginate oligosaccharide and cellooligosaccharide (example 1) 33+33+33ppm, root irrigation, 100 mL/plant
Chitosan oligosaccharide, brown alginate oligosaccharide and pectin oligosaccharide (example 2) 33+33+33ppm, root irrigation, 100 mL/plant
Chitosan oligosaccharide, brown alginate oligosaccharide and xylo-oligosaccharide (example 3) 33+33+33ppm, root irrigation, 100 mL/plant
(2) Investigation method
1) Field management data
The growth environment, soil and the like of greenhouse crops are kept consistent.
2) The effects of the chemical on the growth and vigor of crops are observed by visual measurement and measurement methods, and the effects of each treatment on the growth, the yield, the fruit quality and the like of crops and the disease control effect are clarified by measuring the plant height, the quality, the yield and the like of the crops.
(1) During the whole period of crop growth, before and after each application, the growth and disease conditions of the crops are observed and recorded, and the plant height and stem thickness of the plants are measured to determine the relative increment.
(2) And in the harvest stage, recording the yield and quality of the fruits, and evaluating the mouthfeel.
(3) Tomato quality measurement: and measuring indexes such as total sugar, total acid and vitamin C content of mature fruits, and the like, and determining the influence of each treatment on the quality of tomatoes.
(4) Yield determination: three ridges (6 rows) of tomatoes in an experimental group and a control group are selected, mature tomatoes are picked, weight measurement and summarization are carried out, and the influence of each treatment on tomato yield is determined.
TABLE 6 influence of treatments on tomato quality and yield
As can be seen from the data in table 6, the total sugar of tomatoes in the experimental group is improved by 4.9% -17.1% compared with that in the control group, the total acid is reduced by 9.2% -18.3%, the maximum increase of vitamin C is 75%, and the yield increase is 2.2% -9.3%. The quality and the yield of the chitosan oligosaccharide, brown alginate oligosaccharide and cellooligosaccharide combined with the pectin oligosaccharide or the xylooligosaccharide treatment group are good, wherein the quality and the yield of the chitosan oligosaccharide, brown alginate oligosaccharide and cellooligosaccharide treatment group are optimal.
The oligosaccharide biological stimulator and the application method thereof provided by the invention have the advantages that the active ingredients of the biological stimulator are composed of different oligosaccharides, the functions of improving stress resistance of crops, promoting growth of the crops and improving quality and yield are realized, the vitality of the crops can be improved, the yield and quality of agricultural products can be improved, and the problems of growth inhibition, overgrowth or weak immunity caused by using a single oligosaccharide are avoided.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.

Claims (9)

1. An oligosaccharide biological stimulant, which is characterized in that the active ingredients comprise a component A, a component B and a component C; the component A is chitosan oligosaccharide, the component B is brown alginate oligosaccharide, and the component C is one or more of pectin oligosaccharide, cellooligosaccharide and xylooligosaccharide in any proportion.
2. The oligosaccharide biological stimulant according to claim 1, wherein the stimulant comprises 1-5wt% of component A, 1-5wt% of component B, 1-5wt% of component C, 1.1-6wt% of auxiliary materials, the balance of solvents and the pH value is 4.0-8.0.
3. An oligosaccharide biological stimulant according to claim 1 wherein the degree of polymerisation of the active ingredients is in the range 2 to 10.
4. An oligosaccharide biological stimulant according to claim 3 wherein the degree of polymerisation of the active ingredient is in the range 4 to 10.
5. A method of using an oligosaccharide biostimulant according to any one of claims 1-4, wherein said biostimulant is sprayed or applied to the roots of a crop plant.
6. The method of using an oligosaccharide-type biostimulant according to claim 5, wherein said biostimulant is used for agricultural crops at an active ingredient concentration of 0.5-200ppm.
7. The method of using an oligosaccharide-type biostimulant according to claim 6, wherein said biostimulant is used for agricultural crops and has a foliar spray active ingredient concentration of 20-200ppm.
8. The method of using an oligosaccharide biostimulant according to claim 6, wherein said biostimulant is used in agricultural crops and the concentration of root-applied active ingredient is 10-120ppm.
9. The method of using an oligosaccharide-based biostimulant according to claim 5, wherein the crop is one of the group consisting of Asteraceae, solanaceae, brassicaceae, gramineae, leguminosae, rosaceae, cucurbitaceae, convolvulaceae, chenopodiaceae, liliaceae, umbelliferae, malvaceae, zingiberaceae, and Nelumbo nucifera.
CN202310797215.XA 2023-06-30 2023-06-30 Oligosaccharide biological stimulant and application method thereof Pending CN116831125A (en)

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