CN111587885A - Bactericidal composition compounded by water-soluble chitosan and benziothiazolinone with specific molecular weight and application thereof - Google Patents
Bactericidal composition compounded by water-soluble chitosan and benziothiazolinone with specific molecular weight and application thereof Download PDFInfo
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- CN111587885A CN111587885A CN202010594676.3A CN202010594676A CN111587885A CN 111587885 A CN111587885 A CN 111587885A CN 202010594676 A CN202010594676 A CN 202010594676A CN 111587885 A CN111587885 A CN 111587885A
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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
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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
- A01N43/72—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms
- A01N43/80—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with nitrogen atoms and oxygen or sulfur atoms as ring hetero atoms five-membered rings with one nitrogen atom and either one oxygen atom or one sulfur atom in positions 1,2
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- Agronomy & Crop Science (AREA)
- Pest Control & Pesticides (AREA)
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- Engineering & Computer Science (AREA)
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- General Health & Medical Sciences (AREA)
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Abstract
The invention belongs to the field of novel compound pesticides, and particularly provides a sterilization composition compounded by water-soluble chitosan and benziothiazolinone with specific molecular weight and application thereof, a series of water-soluble chitosan with specific viscosity-average molecular weight of 10-300 kDa is prepared by adopting specific enzyme, and then 10-60 kDa target chitosan with outstanding antibacterial effect is selected as a compound synergistic component according to an antibacterial test result, the weight percentage of active ingredients in the compound sterilization composition is 2-50%, and the weight ratio of the water-soluble chitosan with specific molecular weight to the benziothiazolinone is 1: 20-5: 1, can be processed into water suspension, microemulsion, soluble powder, wettable powder and water dispersible granules, has good bacteriostasis and sterilization effects on a plurality of plant pathogenic bacteria such as ginger plague pathogenic bacteria, cucumber hornspot pathogenic bacteria and the like under the effective concentration, reduces the dosage of benziothiazolinone, realizes the decrement and the synergy of chemical agents, can promote the growth of crops, and improves the resistance of the crops, the yield and the safety of agricultural products.
Description
Technical Field
The invention relates to the technical field of novel pesticide compositions, and particularly provides a bactericidal composition compounded by water-soluble chitosan and benziothiazolinone with specific molecular weight and application thereof.
Background
Chitosan (chitosan), also known as chitosan, is the only basic polysaccharide in nature and has a wide source. The chitosan is a cationic biological polysaccharide formed by connecting glucosamine and N-acetylglucosamine through beta-1, 4 glycosidic bonds, and the degree of deacetylation is generally more than 85%. Domestic and foreign researches show that the chitosan can induce crops to generate systemic resistance and also has obvious inhibition or killing effects on various plant pathogenic microorganisms, and the bacteriostatic ability of the chitosan is closely related to pathogenic bacteria species, the molecular weight of the chitosan, the deacetylation degree and the like. Since the 21 st century, chitosan and its derivatives have been regarded as novel antibacterial agents, and research has found that the possible antibacterial mechanisms mainly include four types: (1) the chitosan (or chitosan oligosaccharide) has free amino with positive charges, and the positive charges are combined with negative charges on the surface of a germ cell membrane to block the proliferation of germs, thereby playing a role in inhibiting bacteria; (2) chitosan (or chitosan oligosaccharide) increases the permeability of germ cell membrane to cause the loss of intracellular substances, which leads to the weakening or death of cell vitality; (3) the high molecular weight chitosan forms a compact outer membrane on the surfaces of germ cells to prevent nutrient substances from entering the cells, so that the cells die; (4) the low molecular weight chitosan (chitosan oligosaccharide) enters the interior of germ cells, is combined with substances such as intracellular chromosomes and the like, interferes with the normal metabolism of the cells, and thus has the bacteriostatic action.
The benziothiazolinone has good control effect on diseases such as plant bacterial angular leaf spot, canker and the like, but the benziothiazolinone bactericide is used alone for a long time, so that the pathogenic bacteria easily generate drug resistance. The chitosan and chitosan oligosaccharide have a synergistic effect when being combined with the bactericides and fungi chemically synthesized, so that the dosage of the bactericides can be effectively reduced, and the drug resistance of the bactericides can be overcome or delayed. Meanwhile, the chitosan can induce crops to generate resistance, and the crop yield is improved. However, chitosan in the traditional concept is only soluble in dilute acid and has a large molecular weight, which greatly limits the application range and effect of the chitosan. Research shows that only when the molecular weight of chitosan is reduced to a certain range, more biological activity can be shown, and the chitosan is more beneficial to absorption and utilization of crops, and when the molecular weight is lower, such as chitosan oligosaccharide, the growth promotion effect is better, but the sterilization effect is inferior. Therefore, how to obtain a combination product of water-soluble chitosan and benziothiazolinone with a specific molecular weight to fill the gap in the field becomes one of the problems to be solved urgently by the technical personnel in the field.
Disclosure of Invention
Aiming at the defects of the technology, the invention provides a sterilization composition compounded by water-soluble chitosan with specific molecular weight and benziothiazolinone and application thereof, the sterilization composition adopts specific enzyme to prepare a series of water-soluble chitosan with specific viscosity-average molecular weight of 10-300 kDa, and then selects target chitosan with 10-60 kDa and outstanding bacteriostasis effect as a compound synergistic component according to the bacteriostasis test result, the weight percentage of the effective components in the compound sterilization composition is 2-50%, and the weight ratio of the water-soluble chitosan with specific molecular weight and benziothiazolinone is 1: 20-5: 1, can be processed into water suspension, microemulsion, soluble powder, wettable powder and water dispersible granules, has good bacteriostasis and sterilization effects on a plurality of plant pathogenic bacteria such as ginger plague pathogenic bacteria, cucumber hornspot pathogenic bacteria and the like under the effective concentration, reduces the dosage of benziothiazolinone, realizes the decrement and the synergy of chemical agents, can promote the growth of crops, and improves the resistance of the crops, the yield and the safety of agricultural products.
Compared with the prior art, the application has the greatest characteristic that the optimal water-soluble chitosan with specific molecular weight is screened out and then compounded with the benziothiazolinone, although the technical scheme of compounding the chitosan oligosaccharide and the benziothiazolinone is adopted in the prior art, the bacteriostatic effect and the biological activity of the chitosan are difficult to satisfy, so that the inventor abandons the acid-soluble chitosan and the chitosan oligosaccharide in the prior art, selects the water-soluble chitosan with specific molecular weight as the raw material, obtains unexpected technical effect, and has better bacteriostatic effect and other biological activity than the acid-soluble chitosan and the chitosan oligosaccharide.
At present, the action mechanisms of chitosan or chitosan oligosaccharide mainly include four, as mentioned above;
the bactericidal action mechanism of benziothiazolinone mainly comprises two mechanisms: (1) destroying the nuclear structure of pathogenic bacteria, resulting in cell exhaustion and death; (2) interfere the metabolism of germ cells, disturb the physiology of germ cells and finally cause death.
Therefore, in the technical scheme of the invention, the water-soluble chitosan with specific molecular weight has the same bacteriostasis or sterilization action mechanism (similar to the benziothiazolinone sterilization mechanism) as chitosan oligosaccharide, and can form a compact outer membrane on the cell surface of pathogenic bacteria to kill the pathogenic bacteria, which is also the supplement of the benziothiazolinone effect, so that the synergistic effect is better.
The specific technical scheme provided by the invention is as follows:
a sterilization composition compounded by water-soluble chitosan and benziothiazolinone with specific molecular weight is disclosed, wherein the water-soluble chitosan and benziothiazolinone with specific molecular weight are effective components, the weight percentage of the effective components is 2% -50%, and the weight ratio of the water-soluble chitosan and benziothiazolinone with specific molecular weight is 1: 20-5: 1;
wherein the water-soluble chitosan with specific molecular weight is selected from chitosan with molecular weight of 10 kDa-60 kDa, and through detection, the antibacterial activity of the chitosan with the molecular weight is obviously superior to that of chitosan oligosaccharide, and the water solubility is obviously superior to that of chitosan with traditional concept, specifically, the contents of the following experimental example 1 and table 1 can be referred to;
on one hand, the bactericidal composition can improve the bacteriostatic and bactericidal activity of the benziothiazolinone on plant pathogenic bacteria, reduce the using amount of chemical agents, and is green, safe and environment-friendly; on the other hand, the function of inducing immunity of the product is increased. The invention has double effects of pesticide and fertilizer, has important significance for agricultural product safety and green agricultural development, and can refer to the related data in table 3 in the following invention contents;
further, the inventor provides that the weight percentage of the effective components is 3.5-15%, and the weight ratio of the specific molecular weight water-soluble chitosan to the benziothiazolinone is 1: 10-4: 1;
in specific application, the optional dosage form is any one of an aqueous suspension agent, a microemulsion, a soluble powder, a wettable powder and a water dispersible granule, and the required auxiliary agents are conventional auxiliary agents in the field, so that the inventor does not need to describe any more; when the bactericidal composition is applied, the bactericidal effect on ginger blast pathogenic bacteria and cucumber angular leaf spot pathogenic bacteria is good when the effective final concentration is below 1.0 ppm.
In conclusion, the invention combines the water-soluble chitosan with specific molecular weight and the benziothiazolinone, not only exerts the special function of the chitosan and increases the immunity inducing function, but also improves the bactericidal activity of the benziothiazolinone to plant pathogenic bacteria, reduces the using amount of the benziothiazolinone, and improves the safety and the yield of agricultural products. Meanwhile, the chitosan with lower dosage can play a role in synergy, so that the problems of large dosage and high cost of the chitosan used alone are avoided, and a theoretical basis is provided for the wide application of the chitosan in the agricultural field.
Detailed Description
The present invention will be described in further detail with reference to the following examples, but it should not be construed that the scope of the above subject matter is limited to the following examples. All the technologies realized based on the above contents of the present invention belong to the scope of the present invention, and the following embodiments are all completed by adopting the conventional prior art except for the specific description.
Example 16% Thiothiazolinone-Water soluble Chitosan aqueous suspension
The contents of the components (by weight percentage) are 3 percent of benziothiazolinone, 3 percent of water-soluble chitosan with specific molecular weight, 2 percent of magnesium aluminum silicate, 0.2 percent of xanthan gum, 5 percent of naphthalenesulfonate dispersant, 5 percent of ethylene glycol and distilled water which is added to make up to 100 percent. Mixing the above materials, grinding with colloid mill for 3-5 hr to make the particle diameter of the material below 5 μm.
Effect test example 1
Screening of bacteriostatic effect of water-soluble chitosan with different molecular weights on plant pathogenic bacteria
Test agents: water-soluble chitosan with molecular weight of 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 200 and 300kDa respectively;
control agents: chitosan oligosaccharide, commercially available acid-soluble chitosan;
test bacteria: ginger bacterial wilt pathogen, cucumber angular leaf spot pathogen;
activating strains: selecting two bacteria of ginger bacterial wilt pathogenic bacteria and cucumber angular leaf spot protobacteria as bacteriostatic objects, adding 50mL of LB seed culture medium (the existing culture medium) into a 250mL triangular flask, inoculating an environment-friendly bacterial slant seed stored at 4 ℃, and culturing at 160r/min at 37 ℃ overnight.
And (3) screening the optimal concentration of bacteria:
adding 1mL of activated bacterial liquid into 9mL of sterile water, sequentially diluting for 6 gradients, uniformly coating 100 mu L of diluted bacterial liquid on a flat plate of a solid LB culture medium, culturing overnight at 37 ℃, and finding out the optimal bacterial liquid concentration for a bacteriostatic test according to the number of bacterial colonies on the flat plate.
And (3) determining the bacteriostatic effect and the minimum bacteriostatic concentration:
respectively preparing LB solid culture media containing chitosan samples with different weight percentages, pouring the plates after fully and uniformly shaking, and marking. With LB medium without any sample as a blank, containing different concentrations of chitosan oligosaccharide and acid-soluble chitosan on the marketLB medium was a positive control. Respectively taking the concentration of 1.5 x 106And uniformly coating 100 mu L of CFU/mL bacterial liquid of ginger bacterial wilt pathogens and cucumber angular leaf spot protobacteria on a flat plate, repeating three treatments for each treatment, culturing at 37 ℃, culturing for 24h, and counting respectively. Comparing the bacteriostatic effect, screening out chitosan with specific molecular weight and good bacteriostatic effect, and determining the minimum bacteriostatic concentration of the chitosan. The results are shown in tables 1 and 2.
The minimum inhibitory concentration mic (minimum inhibitory concentration) refers to the minimum drug concentration that can inhibit a certain microorganism from obviously increasing after incubation for 24 hours in a specific environment, namely the minimum inhibitory concentration.
Minimum bactericidal concentration, mbc (minimum bacterial concentration), is the minimum drug concentration required to kill 99.9% of the test microorganisms.
TABLE 1 inhibition effect of water-soluble chitosan of different molecular weights
As can be seen from Table 1, when the final concentration is 0.03% or 0.04%, the water-soluble chitosan with different molecular weights has larger difference in bacteriostasis effect, and compared with acid-soluble chitosan (chitosan in the traditional concept) and chitosan oligosaccharide, the water-soluble chitosan with the molecular weight of 10 kDa-60 kDa has better inhibition effect on the ginger blast disease protobacteria and cucumber angular leaf spot pathogenic bacteria.
TABLE 2 minimum inhibitory concentration and minimum bactericidal concentration of water-soluble chitosan with different molecular weights
The minimum inhibitory concentration and the minimum bactericidal concentration of the water-soluble chitosan with the molecular weight of 10 kDa-60 kDa are shown in Table 2, and the water-soluble chitosan with the molecular weight of 50kDa with the best inhibitory effect is shown.
Effect test example 2
Specific molecular weight water-soluble chitosan and benziothiazolinone composition sterilizing effect
Test agents: water-soluble chitosan with molecular weight of 50 kDa: benziothiazolinone ═ 1: 10
Control agents: zhongshengmycin, acid-soluble chitosan and benziothiazolinone composition and chitosan oligosaccharide and benziothiazolinone composition
Test bacteria: pathogenic bacteria of ginger blast and angular leaf spot of cucumber
And (3) determination of the bactericidal effect: respectively preparing LB solid culture media containing compositions with different concentrations, pouring the plates after fully and uniformly shaking, and marking. LB medium without any sample was used as a blank control, and LB medium with different concentrations of benziothiazolinone (see Table 3) and zhongshengmycin was used as a positive control; applying 2 μ L of bacterial solution of ginger bacterial wilt pathogenic bacteria or cucumber angular leaf spot primary bacteria with appropriate concentration onto the plate, wherein the bacterial count at each point is about 1.5 × 104CFU, forming bacterial plaque with the diameter of 5-8 mm. Two times of each treatment are repeated, the culture is carried out at 37 ℃, the culture is carried out for 48 hours, and the bactericidal effect is compared according to the growth condition of bacterial plaques. The results are shown in Table 3.
TABLE 3 fungicidal Effect of specific molecular weight Water-soluble Chitosan and Thiazone compositions
TABLE 4 synergistic interaction of water-soluble chitosan and benziothiazolinone at specific molecular weights
As can be seen from tables 3 and 4, the bactericidal effect and the synergistic effect of the composition of the water-soluble chitosan and the benziothiazolinone with specific molecular weight are obvious, and the MIC/MBC is reduced by more than 20% compared with the benziothiazolinone used alone, which is beneficial to reducing the dosage of pesticide.
Claims (4)
1. A sterilization composition compounded by water-soluble chitosan and benziothiazolinone with specific molecular weight is characterized in that: the weight percentage of the effective components is 2% -50%, and the weight ratio of the specific molecular weight water-soluble chitosan to the benziothiazolinone is 1: 20-5: 1;
wherein the water-soluble chitosan with specific molecular weight is selected from chitosan with molecular weight of 10 kDa-60 kDa.
2. The bactericidal composition compounded by the specific molecular weight water-soluble chitosan and the benziothiazolinone according to claim 1, which is characterized in that: the weight percentage of the effective components is 3.5-15%, and the weight ratio of the specific molecular weight water-soluble chitosan to the benziothiazolinone is 1: 10-4: 1.
3. the bactericidal composition compounded by the specific molecular weight water-soluble chitosan and the benziothiazolinone according to claim 1, which is characterized in that: the formulation of the composition is one of aqueous suspension, microemulsion, soluble powder, wettable powder and water dispersible granules.
4. The application of the bactericidal composition compounded by the specific molecular weight water-soluble chitosan and the benziothiazolinone in the claim 1 in the prevention and control of ginger blast pathogenic bacteria and cucumber angular leaf spot pathogenic bacteria.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102835402A (en) * | 2012-05-29 | 2012-12-26 | 陕西西大华特科技实业有限公司 | Bactericidal composition containing benziothiazolinone and chitosan oligosaccharide |
| CN104798789A (en) * | 2014-01-28 | 2015-07-29 | 中国科学院大连化学物理研究所 | Broad-spectrum oligosaccharide compound bactericide and use thereof |
| CN107787988A (en) * | 2017-09-28 | 2018-03-13 | 江苏明德立达作物科技有限公司 | A kind of chitosan oligosaccharide and benziothiazolinone oil-suspending agent and preparation method thereof |
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- 2020-06-24 CN CN202010594676.3A patent/CN111587885A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102835402A (en) * | 2012-05-29 | 2012-12-26 | 陕西西大华特科技实业有限公司 | Bactericidal composition containing benziothiazolinone and chitosan oligosaccharide |
| CN104798789A (en) * | 2014-01-28 | 2015-07-29 | 中国科学院大连化学物理研究所 | Broad-spectrum oligosaccharide compound bactericide and use thereof |
| CN107787988A (en) * | 2017-09-28 | 2018-03-13 | 江苏明德立达作物科技有限公司 | A kind of chitosan oligosaccharide and benziothiazolinone oil-suspending agent and preparation method thereof |
Non-Patent Citations (3)
| Title |
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| 何日安等: "低分子量壳聚糖对植物病原菌的抑制作用研究 ", 《安徽农业科学》 * |
| 欧阳海燕等: "水溶性壳聚糖生物活性及其制备技术的研究近况", 《抗感染药学》 * |
| 马鹏鹏等: "壳聚糖对植物病原细菌的抑制作用研究", 《天然产物研究与开发》 * |
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Application publication date: 20200828 |