阅读说明：本技术 氯唑沙宗在制备用于防治由植物病原菌引起的植物病害的杀菌剂中的应用 (Application of chlorzoxazone in preparing bactericide for preventing and treating plant diseases caused by phytopathogen ) 是由 宋丽敏 王光远 梁文星 刘美洁 范慧珍 于 2021-01-26 设计创作，主要内容包括：本发明提供了氯唑沙宗在制备用于防治由植物病原菌引起的植物病害的杀菌剂中的应用。本发明通过测试氯唑沙宗对番茄灰霉病菌和尖孢镰刀菌番茄专化型的菌丝生长、孢子萌发和芽管伸长的抑制实验,证明氯唑沙宗对番茄灰霉病菌和尖孢镰刀菌番茄专化型具有较强的抑制作用。传统的化学农药防治植物病原菌引起的植物病害对环境污染大,农药残留直接威胁着食品安全。氯唑沙宗作为杀菌剂,具有高效和低毒的优点,适合于植物病害化学防治的要求。氯唑沙宗是一种环境友好的小分子化合物,对非靶标生物及人畜安全,能够保证农产品及果蔬的高品质,符合可持续发展的要求,其研究和市场应用前景广阔。(The invention provides application of chlorzoxazone in preparing a bactericide for preventing and treating plant diseases caused by phytopathogens. According to the inhibition experiment of chlorzoxazone on the growth of hyphae, spore germination and sprout elongation of tomato specialization types of botrytis cinerea and fusarium oxysporum, the invention proves that chlorzoxazone has a strong inhibition effect on the tomato specialization types of botrytis cinerea and fusarium oxysporum. The traditional chemical pesticide for preventing and treating plant diseases caused by phytopathogens has great environmental pollution, and pesticide residues directly threaten food safety. The chlorzoxazone as bactericide has the advantages of high efficiency and low toxicity, and is suitable for the requirement of chemical prevention and control of plant diseases. Chlorzoxazone is an environment-friendly small molecular compound, is safe to non-target organisms, people and livestock, can ensure the high quality of agricultural products, fruits and vegetables, meets the requirement of sustainable development, and has wide research and market application prospects.)

1. The application of chlorzoxazone in preparing bactericide for preventing and treating plant diseases caused by plant pathogenic bacteria.

2. The use of chlorzoxazone according to claim 1 in the preparation of a fungicide for the control of plant diseases caused by phytopathogens, characterized in that: the plant pathogenic bacteria are tomato botrytis cinerea and fusarium oxysporum tomato specialization types.

3. The use of chlorzoxazone according to claim 1 or 2 in the preparation of a fungicide for the control of plant diseases caused by phytopathogens, characterized in that: the effective using concentration of the chlorzoxazone in the bactericide is 0.1 mM-1.0 mM.

4. The use of chlorzoxazone according to claim 3 in the preparation of a fungicide for the control of plant diseases caused by phytopathogens, characterized in that: the inhibition rate of chlorzoxazone on the growth of tomato special type hyphae of botrytis cinerea and fusarium oxysporum is dose-dependent.

5. The use of chlorzoxazone according to claim 4 in the preparation of a fungicide for the control of plant diseases caused by phytopathogens, characterized in that: when the concentration of chlorzoxazone in the bactericide is 1.0 mM, the inhibition rate of the chlorzoxazone on the growth of tomato special type hyphae of botrytis cinerea and fusarium oxysporum is more than 98.4%.

6. The use of chlorzoxazone according to claim 3 in the preparation of a fungicide for the control of plant diseases caused by phytopathogens, characterized in that: the inhibition rate of chlorzoxazone on the spore germination of botrytis cinerea and fusarium oxysporum tomato specialization type is dose-dependent.

7. The use of chlorzoxazone according to claim 6 in the preparation of a fungicide for the control of plant diseases caused by phytopathogens, characterized in that: when the concentration of chlorzoxazone in the bactericide is 0.5 mM, the inhibition rate of chlorzoxazone on the germination of tomato botrytis cinerea and fusarium oxysporum tomato specialized spores is over 96%.

8. The use of chlorzoxazone according to claim 3 in the preparation of a fungicide for the control of plant diseases caused by phytopathogens, characterized in that: the inhibition rate of chlorzoxazone on the elongation of tomato special germ tube of botrytis cinerea and fusarium oxysporum is dose-dependent.

9. The use of chlorzoxazone according to claim 8 in the preparation of a fungicide for the control of plant diseases caused by phytopathogens, characterized in that: when the concentration of chlorzoxazone in the bactericide is 0.5 mM, the inhibition rate of the chlorzoxazone on the elongation of the tomato special type of botrytis cinerea and fusarium oxysporum tomato exceeds 97.5%.

10. The use of chlorzoxazone according to claim 1 in the preparation of a fungicide for the control of plant diseases caused by phytopathogens, characterized in that: the plant is selected from fructus Lycopersici Esculenti, fructus Solani Melongenae, Capsici fructus, fructus Cucumidis Sativi, fructus Vitis Viniferae, and strawberry.

Technical Field

The invention belongs to the technical field of agricultural biology, relates to a bactericidal active compound, and particularly relates to an application of chlorzoxazone in preparing a bactericide for preventing and treating plant diseases caused by plant pathogenic bacteria.

Background

The tomato gray mold is a common fruit and vegetable disease caused by botrytis cinerea infection and is also one of the most harmful plant diseases. In addition to tomato, botrytis cinerea can also damage eggplant, pepper, cucumber, grape, strawberry and other important economic crops, and can cause fruit and vegetable botrytis. The disease may occur not only in the growing season of the host plant but also during the storage of the agricultural product, and thus is seriously harmful. Especially, due to the large-area popularization of the facility cultivation mode in recent years, the pathogen infection sources are wide, the propagation speed is high, the gray mold of fruits and vegetables is more and more serious, and the occurrence and the prevalence of the gray mold often cause serious economic loss. The occurrence of gray mold of tomatoes in northern facilities is generally reported to be serious, the economic loss caused by general years can reach 20% -30%, and more than 50% of serious years can even be no more than absolutely harvested.

The pathogen of the plant fusarium wilt is fusarium oxysporum, which is a soil-borne pathogenic fungus distributed worldwide, has a wide host range, and can cause the fusarium wilt of more than 100 fusarium wilt of melons, solanaceae, bananas, cotton, leguminous, flowers and the like. After the host is infected by fusarium, the symptoms are diversified, which generally causes the browning of vascular bundles, the wilting and withering of plants, the rotting of bulbs and roots, the weak growth of plants and the like. Especially, fusarium oxysporum tomato specialization causes tomato blight, and seriously threatens tomato production. Fusarium wilt is also an important fungal soil-borne disease causing harm to agricultural production in recent years, and has been generated in many areas, particularly in crop areas with higher serious influence on crop yield and economic benefit.

The botrytis cinerea and fusarium oxysporum have high propagation speed and generate a large number of conidia, so that the conidia are easy to mutate to generate drug resistance, which not only reduces the control effect, but also increases the dosage of the medicament. The excessive use of the pesticide not only causes the pesticide residue of agricultural products to exceed the standard and endanger the health of consumers, but also causes environmental pollution and damages ecological balance and influences the sustainable and healthy development of agriculture. Therefore, there is a need to develop a bactericide which is highly efficient, low toxic, low in residue and environmentally friendly, and to apply it to production practices. Agriculture in China is to be developed towards a healthy and sustainable direction, and the development of bactericide products is oriented to solving the problems of resistance, plant immunity enhancement, safety, high efficiency, good environmental compatibility and the like.

Chlorzoxazone, chemical name 5-chloro-2-benzoxazolone; molecular formula C₇H₄ClNO₂(ii) a The english name chlorezoxazone; CAS number 95-25-0; molecular weight 169.57.

The molecular structural formula is:

chlorzoxazone is a centrally acting muscle relaxant, and is used for treating acute and chronic soft tissue (muscle and ligament) sprain, contusion, post-exercise muscle soreness, muscle spasm caused by central neuropathy and chronic fasciitis.

At present, no report about the inhibition effect of chlorzoxazone on agricultural plant pathogenic bacteria exists at home and abroad.

Disclosure of Invention

The invention aims to provide application of chlorzoxazone in preparing a bactericide for preventing and treating plant diseases caused by phytopathogens. Toxicity measurement proves that chlorzoxazone has good inhibitory activity on botrytis cinerea and fusarium oxysporum tomato specialization types. The chlorzoxazone is used as a bactericide, has high efficiency and low toxicity, and is suitable for the requirements of chemical prevention and control of plant diseases.

In order to realize the purpose of the invention, the invention adopts the following technical scheme to realize:

the invention provides application of chlorzoxazone in preparing a bactericide for preventing and treating plant diseases caused by phytopathogens.

Further: the plant pathogenic bacteria are tomato botrytis cinerea and fusarium oxysporum tomato specialization types.

Further: the effective using concentration of the chlorzoxazone in the bactericide is 0.1 mM-1.0 mM.

Further: the inhibition rate of chlorzoxazone on the growth of tomato special type hyphae of botrytis cinerea and fusarium oxysporum is dose-dependent.

Further: when the concentration of chlorzoxazone in the bactericide is 1.0 mM, the inhibition rate of the chlorzoxazone on the growth of tomato special type hyphae of botrytis cinerea and fusarium oxysporum is more than 98.4%.

Further: the inhibition rate of chlorzoxazone on the spore germination of botrytis cinerea and fusarium oxysporum tomato specialization type is dose-dependent.

Further: when the concentration of chlorzoxazone in the bactericide is 0.5 mM, the inhibition rate of chlorzoxazone on the germination of tomato botrytis cinerea and fusarium oxysporum tomato specialized spores is over 96%.

Further: the inhibition rate of chlorzoxazone on the elongation of tomato special germ tube of botrytis cinerea and fusarium oxysporum is dose-dependent.

Further: when the concentration of chlorzoxazone in the bactericide is 0.5 mM, the inhibition rate of the chlorzoxazone on the elongation of the tomato special type of botrytis cinerea and fusarium oxysporum tomato exceeds 97.5%.

Further: the plant is selected from fructus Lycopersici Esculenti, fructus Solani Melongenae, Capsici fructus, fructus Cucumidis Sativi, fructus Vitis Viniferae, and strawberry.

Compared with the prior art, the invention has the advantages and the technical effects that: at present, chemical pesticides are mainly used for preventing and treating botrytis cinerea and tomato fusarium wilt. The pesticide bactericide has the problem of pesticide residue, and directly threatens the environment and food safety of human beings. Therefore, the development of pollution-free and pollution-free green bactericides is an important prevention and control method in agricultural production. Many green bactericides have been developed and widely used in production, and have achieved remarkable effects. Indoor toxicity measurement proves that chlorzoxazone has good inhibitory activity on botrytis cinerea and fusarium oxysporum tomato specialization types. And chlorzoxazone is an environment-friendly micromolecular compound, is safe to non-target organisms, people and livestock, can ensure the high quality of agricultural products, fruits and vegetables, meets the requirement of sustainable development, and has wide research and market application prospects.

Drawings

FIG. 1 shows the results of experiments on the inhibition of growth of botrytis cinerea hyphae by chlorzoxazone in the present invention;

FIG. 2 is the result of the experiment of inhibiting the growth of fusarium oxysporum tomato specialized hyphae with chlorzoxazone in the present invention;

FIG. 3 is the results of experiments on inhibition of germination of tomato Botrytis cinerea and Fusarium oxysporum tomato obligate conidia by chlorzoxazone in the present invention;

FIG. 4 is the experimental result of inhibition of elongation of the germ tube of Botrytis cinerea by chlorzoxazone according to the present invention;

FIG. 5 is the experimental result of the inhibition of the elongation of tomato specialized germ tube of Fusarium oxysporum by chlorzoxazone of the present invention.

Detailed Description

The technical solution of the present invention is further described in detail below with reference to the accompanying drawings and specific embodiments.

Example 1

First, experimental material

1. Chlorzoxazone was purchased from Sigma and stored at 4 ℃ until use.

2. In the experiment, the specialization type of botrytis cinerea and fusarium oxysporum tomatoes are stored in a refrigerator at 4 ℃, and the culture medium is a potato (potato) glucose culture medium (PDA for short). The PDA preparation method comprises the following steps: cleaning potato, peeling, weighing 200 g, cutting into small pieces, adding 1000 ml distilled water, boiling for 20 min, filtering with gauze, adding 20g agar and 20g glucose into the filtrate, stirring, adding water to 1000 ml, subpackaging, sterilizing at 121 deg.C for 20 min, and cooling.

3. In this experiment, yeast extract peptone glucose medium (YPD for short) was used for determining spore germination and sprout tube elongation of plant pathogenic bacteria. The YPD preparation method comprises the following steps: 10 g of yeast extract, 20g of peptone, 20g of glucose and 1000 ml of distilled water were weighed out, respectively, at natural pH. Subpackaging, sterilizing at 121 ℃ for 20 minutes, and cooling for later use.

Biocontrol experiment of dithizozone on tomato botrytis cinerea and fusarium oxysporum tomato specialization types

1. Inhibition of growth of plant pathogenic bacteria hyphae by chlorzoxazone

The experimental method comprises the following steps: respectively carrying out activation culture on the botrytis cinerea and fusarium oxysporum tomato specialization types on a PDA (personal digital assistant) plate at 25 ℃ for about 5 days for later use; heating PDA culture medium to melt, cooling to about 50 deg.C, adding chlorzoxazone at different amounts to obtain culture medium containing 0, 0.5, 0.8 and 1.0 mM medicinal liquid, and cooling in culture dish; using a puncher to respectively punch fungus cakes (with the diameter of 0.60 cm) on the edges of tomato special type bacterial colonies of botrytis cinerea and fusarium oxysporum, picking the bacterial colonies to the center of a medicine-containing flat plate by using a sterile inoculating needle, and then placing the culture dish in an incubator (25 ℃) for culture.

And observing and measuring the growth condition of the hyphae in time after treatment. When the drug-free control plates were substantially full, the diameter was measured using the crosshatch method and the data was processed, and the inhibition rate was calculated and photographed. The inhibition ratio (%) (diameter of control newly grown hyphae-diameter of treated newly grown hyphae)/diameter of control newly grown hyphae × 100. Each treatment was repeated 3 times.

The bacteriostatic effect of chlorzoxazone on the growth of tomato dedicated hyphae of botrytis cinerea and fusarium oxysporum is shown in fig. 1 and 2. From fig. 1 and fig. 2, it can be seen that, as the concentration of chlorzoxazone increases, the inhibition effect on two plant pathogenic bacteria is gradually improved, i.e. the inhibition rate on the growth of tomato obligate hyphae of botrytis cinerea and fusarium oxysporum is dose-dependent. At 1.0 mM chlorzoxazone, there was essentially no growth of hyphae from both phytopathogens.

Table 1 shows the inhibition rate of chlorzoxazone on the growth of hyphae of the test pathogenic fungi. As shown in Table 1, when the concentration of chlorzoxazone is 0.8 mM, the inhibition rate to two plant pathogenic bacteria is above 88.7%. When the concentration of chlorzoxazone is 1.0 mM, the inhibition rate on botrytis cinerea reaches 98.4%, and the inhibition rate on fusarium oxysporum tomato specialization type reaches 98.5%, which indicates that chlorzoxazone has obvious inhibition effect on the growth of hyphae of two pathogenic bacteria.

TABLE 1 inhibition rate of chlorzoxazone against the growth of pathogenic bacteria hyphae

2. Inhibition of spore germination by plant pathogenic bacteria by chlorzoxazone

The experimental method comprises the following steps: inoculating the two pathogenic bacteria on a PDA plate respectively to culture until spore is produced; collecting spores and adjusting the concentration of molecular spores to about 6.2X 10 using YPD liquid medium⁵Per ml; adding chlorzoxazone with different masses (concentration 0, 0.1, 0.3 and 0.5 mM) into the conidium suspension respectively; the inhibition effect of chlorzoxazone on the germination of two fungal spores is detected by a concave slide method (50 ul of spore suspension is respectively added into a sterilized concave slide, the spore suspension is subjected to moisturizing culture at 26 ℃, when the germination rate of control conidia reaches 90%, the conidia germination rate of chlorzoxazone treatment with different concentrations is detected, and the inhibition rate of a medicament on the conidia germination is calculated).

The experimental result is shown in figure 3, and the inhibition rate of conidium germination of two tested pathogenic fungi is obviously improved along with the increase of the content of chlorzoxazone. When the content is 0.5 mM, the germination of two conidia can be basically inhibited, which shows that the chlorzoxazone has stronger inhibiting effect on the germination of botrytis cinerea and fusarium oxysporum tomato specialized spores.

3. Influence of chlorzoxazone on elongation of botrytis cinerea germ tubes

The experimental method comprises the following steps: and (3) detecting by adopting the same concave slide method, culturing for 6-10 h, observing and measuring the elongation condition of the bud tube by using a microscope, and calculating the inhibition rate of the medicament on the elongation of the bud tube. Inhibition (%) (control sprout tube length-treated sprout tube length)/control sprout tube length × 100. Each treatment was repeated 3 times.

The experimental result is shown in fig. 4, and at the same time point, the inhibition rate of the elongation of the germ tube is gradually improved along with the increase of the concentration of chlorzoxazone. When the chlorzoxazone is 0.5 mM, the elongation of the botrytis cinerea germ tube is basically inhibited.

Table 2 summarizes the inhibition rate of chlorzoxazone on the elongation of the botrytis cinerea germ tubes, and the inhibition rate of chlorzoxazone on the elongation of the botrytis cinerea germ tubes can reach 98.9% when the concentration of the drug is 0.5 mM.

TABLE 2 inhibition rate of chlorzoxazone (0.5 mM) on Botrytis cinerea germ tube elongation

4. Influence of chlorzoxazone on elongation of fusarium oxysporum tomato specialized germ tube

The experimental method comprises the following steps: and (3) detecting by adopting the same concave slide method, measuring the elongation condition of the bud tube after culturing for 3-9 h, and calculating the inhibition rate of chlorzoxazone on the elongation of the bud tube by adopting the same method.

As shown in FIG. 5, at the same time point, the inhibition rate of the growth of the germ tube is higher with the increase of the concentration of chlorzoxazone, namely, the inhibition rate of the growth of the germ tube of fusarium oxysporum is dose-dependent, and when the concentration of chlorzoxazone is 0.5 mM, the growth of the germ tube of fusarium oxysporum is basically inhibited.

Table 3 summarizes the inhibition rate of chlorzoxazone on fusarium oxysporum tomato specialized germ tube elongation. As can be seen from Table 3, when the concentration of chlorzoxazone is 0.5 mM, the elongation inhibition rate of the tomato specialized fusarium oxysporum germ cells can reach 99.8%.

TABLE 3 inhibition rate of chlorzoxazone (0.5 mM) on elongation of fusarium oxysporum tomato specialized germ tubes

The above examples are only intended to illustrate the technical solution of the present invention, but not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes may be made and equivalents may be substituted for elements thereof; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions.