Chitin derivative-based composite biological regulator and preparation method thereof

Abstract

The invention discloses a composite biological regulator based on a chitin derivative and a preparation method of the composite biological regulator. The composite biological regulator comprises the following active components: the chitin derivative, procymidone and valerian bacteria amine, the coating also comprises an auxiliary agent and water. Wherein the mass ratio content of the active ingredients in the composite biological regulator is 5.5-15%; the mass ratio content of the auxiliary agent in the composite biological regulator is 0.8-2.5%; the rest part of the composite biological regulator is the water; the chitin derivative is chitosan and / or chitosan oligosaccharide; according to the present invention, the chitin derivative is adopted as the main active component, and the component is derived from the natural plant, such that the good biodegradability is provided so as to reduce the environmental pollution caused by the chemical regulator. Meanwhile, procymidone and valerian bacteria amine are used as auxiliary active components and have a synergistic effect with the chitin derivative, so that the disease prevention and pest control effects of the regulator are enhanced, a relatively low dosage is kept, and potential risks to the environment and human bodies are reduced.

Description

Technical Field

[0001] The present invention relates to a plant growth regulator, and particularly to a composite biological regulator based on chitin derivatives and a preparation method thereof. Background Art

[0002] In agricultural production, plant diseases and pests are one of the important factors restricting crop yield and quality. Although traditional chemical regulators can control diseases and pests to a certain extent, long-term use is likely to cause diseases and pests to develop drug resistance, and pose potential threats to the environment and human health.

[0003] Therefore, there is an urgent need for a composite biological regulator based on chitin derivatives and a preparation method thereof to solve the problems in the prior art. Summary of the Invention

[0004] The present invention overcomes the deficiencies of the prior art and provides a composite biological regulator based on chitin derivatives and a preparation method thereof.

[0005] To achieve the above object, the technical solution adopted by the present invention is: a composite biological regulator based on chitin derivatives, wherein the active ingredients of the composite biological regulator include: chitin derivatives, procymidone, and valifenalate; the composite biological regulator further includes an auxiliary agent and water;

[0006] Among them, the mass ratio content of the active ingredients in the composite biological regulator is 5.5-15%;

[0007] The mass ratio content of the auxiliary agent in the composite biological regulator is 0.8-2.5%;

[0008] The rest of the composite biological regulator is the water.

[0009] In a preferred embodiment of the present invention, the chitin derivatives are chitosan and / or chito-oligosaccharide, and the auxiliary agent is one or both of ammonia water and NaOH.

[0010] In a preferred embodiment of the present invention, the mass ratio of the chitin derivatives, procymidone, and valifenalate is 6-7:1-2:1-2.

[0011] In a preferred embodiment of the present invention, the composite biological regulator further includes a surfactant, and the mass ratio content of the surfactant in the composite biological regulator is 0.1-0.5%.

[0012] In a preferred embodiment of the present invention, the composite biological regulator is applicable to the prevention and control of plant diseases and pests, and inhibits plant diseases caused by fungi and pathogenic bacteria.

[0013] In a preferred embodiment of the present invention, a method for preparing a composite biological regulator based on chitin derivatives is proposed. To prepare the composite biological regulator, the following steps are included:

[0014] S1. Mix chitin derivatives, procymidone, and valifenalate evenly in a predetermined ratio to obtain an active ingredient mixture.

[0015] S2. Add a predetermined amount of auxiliary agents and surfactants to the active ingredient mixture and stir to fuse the active ingredients with the auxiliary agents.

[0016] S3. Gradually add water to the mixture obtained in step S2 while stirring until a uniform suspension or solution is formed to obtain the composite biological regulator.

[0017] In a preferred embodiment of the present invention, in step S3, heat is applied while adding water to keep the temperature of the mixture at 30 - 50 °C to accelerate the dissolution and uniform mixing of each component.

[0018] In a preferred embodiment of the present invention, the surfactant is selected from one or a mixture of sodium dodecyl sulfate, sodium lauryl sulfate, and coconut oil amide propyl amine oxide.

[0019] In a preferred embodiment of the present invention, the usage methods of the composite biological regulator include, but are not limited to, spraying, irrigation of roots, seed soaking, and dipping of roots.

[0020] The present invention solves the defects existing in the background technology and has the following beneficial effects:

[0021] (1) In the present invention, chitin derivatives are used as the main active ingredient, and this ingredient is derived from natural plants and has good biodegradability, thereby reducing the environmental pollution caused by chemical regulators. At the same time, procymidone and valifenalate, as auxiliary active ingredients, act synergistically with chitin derivatives, enhancing the disease and pest control effect of the regulator while maintaining a low dosage and reducing the potential risks to the environment and human body.

[0022] (2) The composite biological regulator provided by the present invention has a significant control effect on plant diseases and pests such as rice, and can effectively inhibit plant diseases caused by fungi and pathogens. Moreover, chitin derivatives have the effect of inducing disease resistance in plants and can enhance the immunity of plants themselves, thus achieving long-term prevention and control of diseases and pests. Detailed implementation mode

[0023] Next, in combination with the embodiments of the present invention, the technical solutions in the embodiments of the present invention will be clearly and completely described. Obviously, the described embodiments are only a part of the embodiments of the present invention, rather than all of the embodiments. All other embodiments obtained by those of ordinary skill in the art based on the embodiments of the present invention without creative efforts belong to the scope of protection of the present invention.

[0024] In the following description, many specific details are set forth in order to fully understand the present invention. However, the present invention may also be implemented in other ways different from those described herein. Therefore, the scope of protection of the present invention is not limited by the specific embodiments disclosed below.

[0025] A composite biological regulator based on chitin derivatives and a preparation method thereof. The active ingredients of the composite biological regulator include: chitin derivatives, procymidone, and valifenalate; the composite biological regulator also includes auxiliaries and water.

[0026] Among them, the mass ratio content of the active ingredients in the composite biological regulator is 5.5 - 15%; preferably 8%.

[0027] The mass ratio content of the auxiliaries in the composite biological regulator is 0.8 - 2.5%; preferably 1.5%.

[0028] The rest of the composite biological regulator is water.

[0029] Preferably, the chitin derivative is chitosan and / or chito-oligosaccharide, and the auxiliaries are one or both of ammonia water and NaOH.

[0030] Here, it is further explained that chitosan is derived from plant endophytes. The specific extraction process includes the following:

[0031] Collect samples from healthy and pollution-free plant tissues (such as roots, stems, leaves, etc.), and perform strict surface disinfection on the collected samples to remove surface microbial contamination. Plants such as rice, wheat, corn, soybeans, and cotton can be selected.

[0032] Using a selective medium, inoculate the disinfected samples onto the medium, and isolate plant endophytes by methods such as culturing, observing, and picking single colonies. Purify the isolated endophytes and identify their species and characteristics by molecular biology methods (such as 16S rDNA sequencing).

[0033] Inoculate the purified endophytes into a suitable medium for scale-up culture to obtain a sufficient amount of bacterial cells. Separate the cultured bacterial cells from the medium by methods such as centrifugation and filtration, and perform drying treatment.

[0034] The cell walls of the dried bacteria are broken by physical methods (such as ultrasonic disruption, high-pressure homogenization, etc.) or chemical methods (such as enzymatic hydrolysis) to release the chitin inside the cells. An extractant such as dilute acid or dilute base is added to the disrupted bacterial suspension, and through treatments such as stirring and heating, the chitin is dissolved or released. Then, the chitin is separated from other components by methods such as centrifugation and filtration.

[0035] The chitin obtained by extraction is purified to remove impurities and by-products therein. Common purification methods include dialysis, chromatography, etc. The purified chitin is placed in a sodium hydroxide solution with a relatively high mass fraction and heated at an appropriate temperature to deacetylate the chitin and convert it into chitosan.

[0036] The deacetylated chitosan is separated from the solution by methods such as centrifugation and filtration and then dried. According to needs, post-treatments such as crushing and sieving are carried out on the dried chitosan to obtain a chitosan product that meets the requirements.

[0037] The process for obtaining chitooligosaccharides includes the following: Select chitinase or chitosanase, dissolve the enzyme in a buffer solution to prepare an enzyme solution, and ensure that it has high enzyme activity and stability.

[0038] The chitosan obtained above is dissolved in an acetic acid solution to prepare a chitosan solution. The enzyme solution is added to the chitosan solution, and the pH value and temperature of the reaction system are adjusted to the optimal reaction conditions of the enzyme.

[0039] Within a certain time, a hydrolysis reaction is carried out. After the reaction ends, the enzyme activity is terminated by methods such as heating and adjusting the pH value. The reaction solution is filtered, concentrated, precipitated, dried, crushed, etc. to obtain a chitooligosaccharide product.

[0040] Preferably, the mass ratio of the chitin derivative, procymidone, and valifenalate is 6 - 7:1 - 2:1 - 2. It is preferably 7:2:1.

[0041] Preferably, the composite biological regulator further includes a surfactant, and the mass ratio content of the surfactant in the composite biological regulator is 0.1 - 0.5%. It is preferably 0.2%.

[0042] Preferably, the composite biological regulator is applicable to the prevention and control of plant diseases and pests and inhibits plant diseases caused by fungi and pathogens. Among them, plants include but are not limited to food crops, cash crops, economic forest trees, Chinese medicinal materials, flowers, or lawns.

[0043] Preferably, a preparation method of a composite biological regulator based on a chitin derivative is proposed for preparing the composite biological regulator, including the following steps:

[0044] S1. Mix chitin derivatives, procymidone, and valifenalate evenly in a predetermined ratio to obtain an active ingredient mixture;

[0045] S2. Add a predetermined amount of auxiliary agent and surfactant to the active ingredient mixture and stir to fuse the active ingredient with the auxiliary agent;

[0046] S3. Gradually add water to the mixture obtained in step S2 while stirring until a uniform suspension or solution is formed to obtain a composite biological regulator.

[0047] Preferably, in step S3, heat is applied while adding water to keep the temperature of the mixture at 30 - 50 °C to accelerate the dissolution and uniform mixing of each component. The temperature is preferably 35 °C.

[0048] Preferably, the surfactant is selected from one or a mixture of sodium dodecyl sulfate, sodium lauryl sulfate, and coconut oil amide propyl amine oxide.

[0049] Preferably, the application methods of the composite biological regulator include, but are not limited to, spraying, irrigation, seed soaking, and root dipping.

[0050] Example 1

[0051] The active ingredients selected are 70 g of chitosan, 20 g of procymidone, and 10 g of valifenalate; the auxiliary agent selected is 15 g of a 75% NaOH solution; then 2 g of sodium dodecyl sulfate surfactant is added. After stirring evenly, gradually add clear water until the total mass reaches 1000 g and stop.

[0052] It should be noted that during the process of adding water, continuous heat treatment is carried out on the mixed solution so that the temperature of the mixed solution always remains at 35 °C.

[0053] Stir the mixed solution continuously for 3 min in a cyclic manner of three circles clockwise and two circles counterclockwise to prepare a composite biological regulator, marked as agent A.

[0054] Example 2

[0055] Use chitinase to prepare an enzyme solution with a concentration of 70%; select the same chitosan as in Example 1 and add it to a 75% acetic acid solution to make a chitosan solution; then mix the enzyme solution and the chitosan solution in a mass ratio of 1:3 to make a chitooligosaccharide solution.

[0056] After standing the chitooligosaccharide solution for 30 min, carry out continuous heat treatment to end the hydrolysis reaction. Finally, perform a series of process steps on the heated chitooligosaccharide solution, including filtration, concentration, precipitation, drying, and pulverization, to obtain chitooligosaccharide.

[0057] The active ingredient is selected as 70 g of chito-oligosaccharide, and the other ingredients remain unchanged, and the mixing process remains unchanged, so as to prepare a compound biological regulator, labeled as Medicament B.

[0058] Experiment on the control effect of plant diseases and pests

[0059] A rice paddy in a certain area is selected, and the rice paddy is evenly divided into three parts by area, and are respectively named: experimental group, control group one, and control group two. It should be noted that during the experiment, a time period with sufficient sunlight needs to be selected, which helps to maximize the effect of the compound biological regulator.

[0060] Among them, no medicament is used for the prevention and control of diseases and pests in the experimental group, and only routine daily management is carried out. The daily management includes: weeding, fertilizing, etc.

[0061] In control group one, Medicament A is used, and Medicament A is diluted at a mass ratio of 1:1000, and then in the form of spraying, the diluted Medicament A is sprayed on the surface of the rice and left to air dry.

[0062] In control group two, Medicament B is used, and Medicament B is also diluted at a mass ratio of 1:1000, and then in the form of spraying, the diluted Medicament B is sprayed on the surface of the rice and left to air dry.

[0063] After that, both control group one and control group two carry out the same daily management as the experimental group.

[0064] On the first day, second day, fourth day, seventh day, tenth day, and fifteenth day after that, record the growth situation and the situation of diseases and pests of the rice. The specific situation is shown in Table 1:

[0065] Table 1

[0066]

[0067] It should be explained that Table 1 shows a record of the growth status (S) and the situation of whether there are diseases and pests (H) of an experimental group and control group one and control group two at different time points (the first day, second day, fourth day, seventh day, tenth day, fifteenth day), as well as their evaluation grades (excellent, good, medium, poor) at these time points.

[0068] Change of evaluation grade:

[0069] Experimental group:

[0070] In the initial stage of the experiment (from the first day to the fourth day), the evaluation grade is "excellent". Starting from the seventh day, the evaluation grade gradually decreases, reaching "medium" on the tenth day and "poor" on the fifteenth day.

[0071] Control group one:

[0072] During the entire experiment period, the evaluation grade remained "excellent" for most of the time, only dropping to "good" on the tenth day, but then rising back to "excellent".

[0073] Control Group 2:

[0074] Similar to Control Group 1, the evaluation grade remained "excellent" for most of the time during the entire experiment period, only dropping to "good" on the fifteenth day.

[0075] Comparison between groups:

[0076] Compared with Control Group 1 and Control Group 2, in the later stage of the experiment, that is, after the seventh day, the evaluation grade of the experimental group dropped more significantly.

[0077] Between Control Group 1 and Control Group 2, the change in the evaluation grade was relatively small, and it remained at the "excellent" level for most of the time.

[0078] Conclusion:

[0079] Experimental Group: Over time, the growth state gradually deteriorated, changing from excellent to medium and finally to poor. This indicates that the growth state of rice without chemical treatment gradually worsened under the influence of pests and diseases.

[0080] Control Group 1: During the entire experiment period, the growth state remained excellent. Even though it dropped slightly on the tenth day, it still remained at the good level. This shows that Chemical A has a good promoting effect on the growth of rice and can effectively control pests and diseases.

[0081] Control Group 2: The growth state was similar to that of Control Group 1, remaining excellent all the time, only dropping slightly to good on the fifteenth day. This shows that Chemical B also has a good promoting effect on the growth of rice and can effectively control pests and diseases.

[0082] Analysis of the situation of pests and diseases (H):

[0083] Experimental Group: The situation of pests and diseases gradually deteriorated, changing from excellent to good and finally to poor. This indicates that the problem of pests and diseases of rice without chemical treatment gradually became serious.

[0084] Control Group 1 and Control Group 2: During the entire experiment period, the situation of pests and diseases remained excellent, and no obvious problems of pests and diseases occurred. This shows that both Chemical A and Chemical B can effectively control the pests and diseases of rice.

[0085] Comparison of the effects of chemicals:

[0086] Both Chemical A and Chemical B showed good effects in controlling the pests and diseases of rice and had a promoting effect on the growth of rice.

[0087] During the experiment, the effects of Chemical A and Chemical B were equivalent, with no obvious differences.

[0088] Through comparing the growth states and pest and disease situations of the experimental group, control group 1, and control group 2 at different time points, the following conclusions are drawn:

[0089] Both medicament A and medicament B can effectively control the pests and diseases of rice and have a promoting effect on the growth of rice. During the experiment, the effects of medicament A and medicament B were comparable and there were no obvious differences.

[0090] Under the influence of pests and diseases, the growth state of the rice without medicament treatment gradually deteriorated and the pest and disease problems gradually became serious.

[0091] It also needs to be explained here that on the 10th and 15th days, the temperature of the test plot changed significantly. Therefore, the sudden changes in the growth states of the rice in control group 1 and control group 2 may be affected by the weather.

[0092] In a preferred embodiment, the composition components of the compound biological regulator further include cyclic lipopeptide and modified chitinase. Cyclic lipopeptide and chitinase are endophytes isolated from disease-resistant crops (plants with good growth states), obtained through fermentation and extraction; and cyclic lipopeptide and chitinase are modified and optimized by using genome mining, heterologous expression technology, and protein engineering technology, and finally are compounded with plant-derived volatile organic compounds (VOCs) to form a synergistic biological regulator.

[0093] Chitin derivatives, cyclic lipopeptide, and modified chitinase as the main active components can directly inhibit the growth of pathogenic bacteria and at the same time induce the systemic resistance of plants themselves. The addition of plant-derived volatile organic compounds (VOCs) further enhances the synergistic effect of the pesticide. In addition, this pesticide has good biodegradability and can reduce the potential risks to the environment and human body.

[0094] Based on the ideal embodiments of the present invention as the inspiration, through the above description, relevant personnel can completely make various changes and modifications without departing from the technical idea of this invention. The technical scope of this invention is not limited to the content in the specification and must be determined according to the scope of the claims.

Claims

1. A composite biological regulator based on chitin derivatives, characterized in that, The active ingredients of the composite biological regulator include: chitin derivatives, procymidone, and valifenalate; the composite biological regulator also includes auxiliaries and water; Among them, the mass ratio content of the active ingredients in the composite biological regulator is 5.5-15%; The mass ratio content of the auxiliaries in the composite biological regulator is 0.8-2.5%; The remaining part of the composite biological regulator is the water.

2. The composite biological regulator based on chitin derivatives according to claim 1, characterized in that: The chitin derivatives are chitosan and / or chito-oligosaccharide, and the auxiliaries are one or both of ammonia water and NaOH.

3. The composite biological regulator based on chitin derivatives according to claim 1, characterized in that: The mass ratio of the chitin derivatives, procymidone, and valifenalate is 6-7:1-2:1-2.

4. The composite biological regulator based on chitin derivatives according to claim 1, wherein: The composite biological regulator also includes a surfactant, and the mass ratio content of the surfactant in the composite biological regulator is 0.1-0.5%.

5. A composite biological regulator based on chitin derivatives according to claim 1, characterized in that: The composite biological regulator is applicable to the prevention and control of plant diseases and insect pests, and inhibits plant diseases caused by fungi and pathogenic bacteria.

6. A preparation method of a composite biological regulator based on chitin derivatives, which is applied to the composite biological regulator described in any one of the above, and is characterized in that, It includes the following steps: S1. Mix the chitin derivatives, procymidone, and valifenalate evenly according to a predetermined ratio to obtain an active ingredient mixture; S2. Add a predetermined amount of auxiliaries and surfactant to the active ingredient mixture, and stir to make the active ingredients and auxiliaries fuse; S3. Gradually add water to the mixture obtained in step S2 while stirring until a uniform suspension or solution is formed to obtain the composite biological regulator.

7. The preparation method of a composite biological regulator based on chitin derivatives according to claim 6, characterized in that: In step S3, heat is applied while adding water to keep the temperature of the mixture at 30-50°C to accelerate the dissolution and uniform mixing of each component.

8. The preparation method of a composite biological regulator based on chitin derivatives according to claim 6, characterized in that: The surfactant is selected from one or more of sodium dodecyl sulfate, sodium lauryl sulfate, and coconut oil amide propylamine oxide for mixed use.

9. The preparation method of a composite biological regulator based on chitin derivatives according to claim 6, characterized in that: The application methods of the composite biological regulator include but are not limited to spraying, root irrigation, seed soaking, and root dipping.

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