CN113016805B - Application of phthalide derivatives in preventing and treating plant virus, killing bacteria, killing insects and killing mites - Google Patents

Abstract

The present invention belongs to the field of pesticide technologyThe phthalide derivatives are compounds shown in a general formula.

Description

Application of Phthalide Derivatives in the Control of Plant Viruses, Sterilization, Insecticide and Acaricide

technical field

The invention relates to the application of phthalide derivatives in controlling plant viruses, sterilizing, killing insects and killing mites, and belongs to the technical field of pesticides.

Background technique

Phthalides are one of the characteristic active components of Umbelliferae, which mainly exist in the genus Celery, Cnidium, True Benedict, Angelica, Peony, and Danggui. In addition, plants of other families, such as Amaryllidaceae, Oleander, Gentianaceae, and secondary metabolites of certain fungi and bacteria also contain phthalide compounds and their derivatives. They are mainly divided into monomeric and dimeric phthalides, which are commonly used as pharmaceuticals in Asia, Europe and North America. The first reports of phthalides appeared in the late 19th century, and they were identified by Ciamician and Silber as the odorous constituents of celery essential oil. In the first half of the twentieth century, phthalides were isolated from the traditional medicines Japanese xiong, angelica, and lavender, which is often used as a food condiment.

The biological activity of hemi-gossypol, a biosynthetic precursor of gossypol, is better than that of gossypol in terms of anti-tobacco mosaic virus (TMV), bactericidal and insecticidal properties, and the activity of hemi-gossypol is the best. Phthalides have similar structures to hemigossypolides, and there are few studies on the pesticide activity of these compounds.

SUMMARY OF THE INVENTION

The purpose of the present invention is to provide the application of phthalide derivatives in controlling plant viruses, sterilizing, killing insects and killing mites. The phthalide derivative is a compound having the structure shown in the following general formula:

wherein, R ¹ , R ² , R ³ and R ⁴ are each independently selected from hydrogen, halogen, C1-C6 alkane, and C1-C6 alkoxy;

R is substituted or unsubstituted phenyl, thiophen-2-yl, and the substituents of the substituted phenyl are each independently selected from one of halogen, C1-C6 hydrocarbyl, and C1-C6 alkoxy or more.

The phthalide derivative of the general formula of the invention has excellent anti-plant virus activity, and can well inhibit tobacco mosaic virus, pepper virus, rice virus, tomato virus, sweet potato virus, potato virus, melon virus and corn dwarf flower Leaf virus, etc., can effectively prevent and control virus diseases of various crops such as tobacco, pepper, rice, tomato, melon and vegetables, grain, vegetables, beans, etc., and is especially suitable for the prevention and control of tobacco mosaic disease.

The phthalide derivative of the general formula of the present invention can be used directly as a plant virus inhibitor, or can be used with an agriculturally acceptable carrier, and can also be used with other anti-plant virus agents such as benzothiadiazole (BTH), thiacyl pyraclostrobin (TDL), 4-methyl-1,2,3-thiadiazole-5-carboxylic acid (TDLA), DL-β-aminobutyric acid (BABA), ribavirin, ningnamycin, phenanthroline Dolizidine alkaloid antorfen, bitriazole compounds XY-13 and XY-30, virus A, salicylic acid, polyhydroxy binaphthaldehyde, amino oligosaccharide form an interaction composition, these compositions are used Some perform synergistic effects, and some perform additive effects.

The phthalide derivatives of the general formula of the present invention show bactericidal activity against the following 14 pathogenic bacteria: cucumber wilt, peanut brown spot, apple ring pattern, tomato early blight, gibberellum wheat, potato late blight, rapeseed fungus Kernel, cucumber Botrytis cinerea, rice sheath blight, Phytophthora capsicum, rice bakanae, wheat sheath blight, corn small spot and watermelon anthracnose.

The phthalide derivative of the general formula of the present invention has the activity of killing aphids, adult mites of Tetranychus cinnabarinus and diamondback moth.

The phthalide derivatives of the general formula of the present invention can be used directly as insecticides, acaricides and fungicides, or can be used with an agriculturally acceptable carrier, and can also be used together with other insecticides, acaricides and fungicides such as pyraclostrobin , chlorfenapyr, etoxazole, pyraclostrobin, pyraclostrobin, etc. are used in combination, and some of these compositions exhibit synergistic effects, and some exhibit additive effects.

Detailed ways

The present invention provides phthalide derivatives, which are compounds represented by the general formula:

wherein, R ¹ , R ² , R ³ and R ⁴ are each independently selected from hydrogen, halogen, C1-C6 alkane, and C1-C6 alkoxy;

R is substituted or unsubstituted phenyl, thiophen-2-yl, and the substituents of the substituted phenyl are independently selected from one of halogen, C1-C6 alkane, and C1-C6 alkoxy one or more.

In the present invention, specific examples of C1-C6 alkyl groups can be, for example, methyl, ethyl, n-propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl base, n-hexyl, etc.

The C1-C6 alkoxy group may be an alkoxy group formed by the above-mentioned specific examples of the alkyl group satisfying the limitation of 1 to 6 carbon atoms.

Specific examples of the halogen may be, for example, fluorine, chlorine, bromine, iodine and the like.

In a preferred embodiment of the present invention, the compound shown in the general formula is selected from one of the compounds shown in the following formula:

3-benzylideneisobenzofuran-1(3H)-one (a);

3-(3-methylbenzylidene)isobenzofuran-1(3H)-one (b);

3-(4-Butylbenzylidene)isobenzofuran-1(3H)-one (c);

3-(4-tert-Butylbenzylidene)isobenzofuran-1(3H)-one (d);

3-(4-Methoxybenzylidene)isobenzofuran-1(3H)-one (e);

3-(3-Chlorobenzylidene)isobenzofuran-1(3H)-one (f);

3-(4-Bromobenzylidene)isobenzofuran-1(3H)-one (g);

3-(2-Bromobenzylidene)isobenzofuran-1(3H)-one (h);

3-(thiophene-2-methylene)isobenzofuran-1(3H)-one (i);

3-benzylidene-5-methoxyisobenzofuran-1(3H)-one (j);

3-benzylidene-6-methylisobenzofuran-1(3H)-one (k);

3-benzylidene-5-chloroisobenzofuran-1(3H)-one (1);

3-Benzylidene-5-fluoroisobenzofuran-1(3H)-one (m).

The present invention provides the application of the above-mentioned phthalide derivatives in anti-plant virus activity.

The phthalide derivatives provided by the invention have excellent anti-plant virus activity, and can well inhibit tobacco mosaic virus, pepper virus, rice virus, tomato virus, sweet potato virus, potato virus, melon virus and corn dwarf mosaic virus Viruses, etc., can effectively prevent and control virus diseases of various crops such as tobacco, pepper, rice, tomato, melon and vegetables, grain, vegetables, beans, etc., especially suitable for the prevention and control of tobacco mosaic disease. The phthalide derivatives represented by the general formula show good anti-tobacco mosaic virus activity.

The phthalide derivatives provided by the present invention can be used directly as plant virus inhibitors, or can be used with agriculturally acceptable carriers, and can also be used with other anti-plant virus agents such as benzothiadiazole (BTH), Amine (TDL), 4-methyl-1,2,3-thiadiazole-5-carboxylic acid (TDLA), DL-β-aminobutyric acid (BABA), ribavirin, ningnamycin, phenanthroindole Lixidine alkaloid antorphine, bitriazole compounds XY-13 and XY-30, virus A, salicylic acid, polyhydroxy binaphthaldehyde, amino oligosaccharide form an interaction composition, these compositions have Some performances are synergistic, and some performances are additive.

From the viewpoint of obtaining higher anti-plant virus activity, the phthalide derivatives described in the present invention are preferably selected from one or more of the following compounds:

3-benzylideneisobenzofuran-1(3H)-one (a);

3-(3-methylbenzylidene)isobenzofuran-1(3H)-one (b);

3-(4-Butylbenzylidene)isobenzofuran-1(3H)-one (c);

3-(4-tert-Butylbenzylidene)isobenzofuran-1(3H)-one (d);

3-(4-Methoxybenzylidene)isobenzofuran-1(3H)-one (e);

3-(3-Chlorobenzylidene)isobenzofuran-1(3H)-one (f);

3-(4-Bromobenzylidene)isobenzofuran-1(3H)-one (g);

3-(2-Bromobenzylidene)isobenzofuran-1(3H)-one (h);

3-(thiophene-2-methylene)isobenzofuran-1(3H)-one (i);

3-benzylidene-5-methoxyisobenzofuran-1(3H)-one (j);

3-benzylidene-6-methylisobenzofuran-1(3H)-one (k);

3-benzylidene-5-chloroisobenzofuran-1(3H)-one (1);

3-Benzylidene-5-fluoroisobenzofuran-1(3H)-one (m).

The present invention also provides a method for anti-plant virus by using the above-mentioned phthalide derivative as a plant virus inhibitor.

The present invention provides the application of the above-mentioned phthalide derivatives in killing phytopathogenic bacteria.

The phthalide derivatives provided by the invention have high phytopathogenic activity, and are especially suitable for causing cucumber wilt, peanut brown spot, apple ring streak, tomato early blight, wheat gibberellum, potato late blight, rape sclerotia, One or more of the pathogenic bacteria of cucumber Botrytis cinerea, rice sheath blight, Phytophthora capsicum, rice bakanae, wheat sheath blight, corn small spot and watermelon anthracnose.

In particular, the phthalide derivatives of the present invention exhibit good bactericidal activity. Among them, the inhibition rate of compounds a and l on apple rot fungus was as high as 80% or more. Compound a and the like showed good phytopathogenic activity against various pathogens such as rape sclerotium, cucumber botrytis cinerea and rice sheath blight, and the inhibitory rate of various bacteria reached more than 80%. Compound 1 has applicability to a variety of bacterial species, the inhibition rate of apple ring striae and wheat sheath blight has reached more than 85%, and the inhibition rate of rape sclerotia has reached 95.7%. The effect is similar to that of commercial chlorothalonil. effect is equivalent.

The phthalide derivatives provided by the invention have high insecticidal and acaricidal activities, and have high activity against one or more of diamondback moth, aphids and adult mites of Tetranychus cinnabarinus.

In particular, in the phthalide derivatives of the present invention, the aphid killing rates of compounds d and f are 80% and 90% respectively under the concentration condition of 600 μg/mL, and the lethal rates of compounds a and i to the adult mites of Tetranychus cinnabarinus are both 90%. , For diamondback moth, almost all compounds have good insecticidal effect. Under the condition of concentration of 200μg/mL, the inhibition rate of compounds a, d, e, f, and i is still as high as 100%, and even when the concentration drops to 10 μg/mL, the lethal rate of a, f is still 60%, 70% %.

The phthalide derivatives provided by the present invention can be used directly as insecticides, acaricides and fungicides, or can be used with an agriculturally acceptable carrier, and can also be used together with other insecticides, acaricides and fungicides such as pyrimidine, When used in combination with chlorfenapyr, etoxazole, pyraclostrobin, pyraclostrobin, etc., some of these compositions exhibit synergistic effects, and some exhibit additive effects.

The present invention also provides a method for killing insects and mites by using the above-mentioned phthalide derivatives as insecticides and acaricides.

The present invention also provides a method for sterilizing by using the above-mentioned phthalide derivatives as phytopathogens.

The following examples and bioassay test results can be used to further illustrate the present invention, but are not meant to limit the present invention.

Embodiment 1: the determination of anti-tobacco mosaic virus activity, the determination procedure is as follows:

1. Virus purification and concentration determination:

According to the Tobacco Mosaic Virus SOP specification compiled by the Laboratory of Elements, Nankai University. After the crude virus extract was centrifuged twice with polyethylene glycol, the concentration was determined, and then refrigerated at 4°C for later use.

2. Compound solution preparation:

After weighing, the original drug was dissolved in DMF to prepare a mother solution of 1×105 μg/mL, and then diluted with an aqueous solution containing 1‰ Tween 80 to the desired concentration; the Ningnanmycin preparation was directly diluted with water.

3. Friction inoculated leaves of the appropriate age of Nicotiana sinensis, rinsed with running water, and the virus concentration was 10 μg/mL. After harvesting, cut them off, cut them in half along the midrib of the leaves, immerse the left and right half leaves in 1‰ Tween water and medicine respectively, take them out after 30 minutes, and keep moisturizing and cultivate under suitable light temperature. 3 times. After 3 days, the number of lesions was recorded, and the control effect was calculated.

4. In vivo protection:

The 3-5 leaf stage Shanxi tobacco with uniform growth was selected, and the whole plant was sprayed, and each treatment was repeated 3 times, and a 1‰ Tween 80 aqueous solution was set as a control. After 24 hours, the leaves were spread with emery (500 mesh), and the virus solution was dipped with a writing brush, rubbed twice along the branch direction on the whole leaf surface, and the palm was supported under the leaf, and the virus concentration was 10 μg/mL. Rinse with running water after inoculation. After 3 days, the number of lesions was recorded, and the control effect was calculated.

5. In vivo therapeutic effect:

The 3-5 leaf stage Shanxi tobacco with uniform growth was selected, and the whole leaves were inoculated with the virus with a brush at a concentration of 10 μg/mL, and washed with running water after inoculation. After the leaves were dried, the whole plant was sprayed, and each treatment was repeated 3 times, and a 1‰ Tween 80 aqueous solution was set as a control. After 3 days, the number of lesions was recorded and the control effect was calculated.

6. In vivo passivation

Select the 3-5 leaf stage Shanxi tobacco with uniform growth, mix the agent with the same volume of virus juice and passivate for 30 minutes, inoculate by friction, the virus concentration is 20 μg/mL, rinse with running water after inoculation, repeat 3 times, set 1‰ Tween 80 aqueous solution control. After 3 days, the number of lesions was counted, and the results were calculated.

Inhibition rate (%)=[(number of dead spots in control-number of dead spots in treatment)/number of dead spots in control]×100%

Table 1 Anti-Tobacco Mosaic Virus (TMV) Activity Test Results

The results of anti-tobacco mosaic virus activity showed that some phthalide compounds such as e, i and m showed good relative inhibition rates in three test modes: in vivo passivation, in vivo treatment, and in vivo protection. Among them, the anti-tobacco mosaic virus activities of compound e (in vivo passivation, therapeutic activity, and in vivo protection three modes were (41.2±4.4%, 34.4±3.5%, 45.1±2.7%, 500 μg/mL), compound i (46.4±4.4%, 34.4±3.5%, 500 μg/mL), respectively. 1.1%, 38.7±0.5%, 42.0±3.2%, 500μg/mL), m (40.7±3.6%, 35.±1.8%, 34.±4.1%, 500μg/mL) antiviral activity and commercial varieties of ribavirin (39.4±1.3%, 37.7±0.9%, 41.2±1.5%, 500 μg/mL).

Embodiment 2: Determination of bactericidal activity, the measurement procedure is as follows:

Take tomato early blight as an example, it can be replaced with other bacteria.

In vitro test method: The tomato early blight was cultivated on PDA medium for 7 days, and a 4cm diameter bacterial dish was prepared at the edge of the colony with a hole punch and inoculated on the PDA medium containing 50 μg/ml and no drug for culture. After 4 days, the colony diameter was measured and compared with the control, the inhibition percentage of the drug was calculated.

Table 2 Test results of in vitro bactericidal activity

Compounds a to m all showed a certain bactericidal activity against Pseudomonas apple, most of them were more than 50%, and the inhibition rates of compounds a and l were as high as 80%. Compound a and the like showed good bactericidal activity against a variety of pathogens such as rape sclerotium, cucumber botrytis cinerea, rice sheath blight, and the inhibition rate of various bacteria reached more than 80%. Compound 1 has the best bactericidal activity, and it has applicability to a variety of bacterial species. The inhibition rate of apple ring grain and wheat sheath blight has reached more than 85%, and the inhibition rate of rape sclerotia has reached 95.7%. The effect is comparable to that of commercial chlorothalonil.

Embodiment 3: Determination of insecticidal and acaricidal activities, the measurement procedure is as follows:

Aphid activity test

The test insects were aphids (Aphis laburni Kaltenbach), a normal population of laboratory faba bean leaf feeding. Weigh the medicine, add 1 mL of DMF to dissolve, add two drops of Tween-20 emulsifier, add a certain amount of distilled water, stir evenly, and prepare a liquid with the required concentration. Immerse the broad bean leaves with aphids (about 60) in the agent for 5 seconds, take out and gently spin dry, use filter paper to absorb the excess agent, then insert the broad bean branch into the absorbent sponge, cover the branch with a glass cover, and seal it with gauze , the results were checked at 96 hours, and each compound was repeated 3 times. In contrast, only the emulsifier and solvent were added to distilled water, and the mixture was stirred evenly.

Activity test of adult mites of Tetranychus cinnabarinus

When the dwarf lettuce bean used for the experiment grows to two true leaves, choose a plant with a relatively neat growth, a leaf area of 4-5 square centimeters, and a plant height of about 10 centimeters. The number of insects per plant is controlled at about 60-100. 24 hours after infestation, chemical treatment was carried out. The chemical treatment adopts the plant dipping method, and the dipping time is 5 seconds. After the plants are removed from the liquid, shake gently to shake off excess liquid, then transfer to a hydroponic tank and place at room temperature. Check the results under binoculars 24 hours after treatment. (Do three parallel experiments and take the average).

Plutella xylostella larvae activity test

The leaf dip method proposed by the International Resistance Action Committee (IRAC) was used. Weigh 2 mg of the drug sample on an analytical balance into a 10 mL small beaker, add 50 μL of dimethylformamide (analytical grade) to dissolve, and add 10 mL of water to make a 200 μg/mL drug solution. Dip the cabbage leaves with straight ophthalmic tweezers for 2-3 seconds and shake off the remaining liquid. 1 tablet each time, 3 tablets in total for each sample. Place them on the processing paper in the order of sample marking. After the liquid was dried, it was put into a marked 10cm long straight tube, the 2nd instar larvae of diamondback moth were inserted, and the mouth of the tube was covered with gauze. The experimental treatment was placed in a standard treatment chamber, and the results were checked after 96 h. Each compound was repeated 3 times. In contrast, only the emulsifier and solvent were added to distilled water, and the mixture was stirred evenly.

Table 3 Insecticidal and acaricidal bioassay activity data

Phthalides have significant effects on killing insects and mites. Compounds d and f have aphid killing rates of 80% and 90% at a concentration of 600 μg/mL, respectively. At a concentration of 600 μg/mL, the compound a and i have a lethality rate of 90% against the adult mites of Tetranychus cinnabarinus, phthalide compounds have an excellent lethality rate against diamondback moth, almost all compounds show good insecticidal activity against diamondback moth, under the condition of concentration of 600μg/mL , except for compound c, the lethality rate of all compounds is as high as 100%. When the concentration is 100 μg/mL, the lethality rates of compounds a, e, d, f, and i to diamondback moth are 90%, 90%, 70%, respectively, 80%. Especially compounds a, f, under the condition of low concentration of 10μg/mL, their killing rate to diamondback moth is still as high as 60% and 70%.

Claims (2)

1. the application of a phthalide derivative in the prevention and treatment of plant viruses, plant pathogenic bacteria, plant pests and mites, wherein, the phthalide derivative is a compound shown in the general formula:

Wherein, R ¹ and R ² are hydrogen and methyl; R ³ is one of hydrogen, methoxy, fluorine and chlorine; R ⁴ is one of hydrogen and methyl; R is substituted or unsubstituted phenyl, thiophen-2-yl, and the substituents of the substituted phenyl are independently selected from methyl, n-butyl, tert-butyl, methoxy, chlorine, and bromine a kind of, The plant virus is tobacco mosaic virus, and the plant pathogenic bacteria are cucumber wilt, peanut brown spot, apple ring pattern, tomato early blight, gibberellum wheat, potato late blight, rape sclerotium, cucumber gray mold, rice grain One or more of the phytopathogenic bacteria of Phytophthora wilt, Phytophthora capsicum, rice bakanae, wheat sheath blight, small corn spot and watermelon anthracnose, and the plant pest mites are diamondback moth, aphids and adult cinnabar mites. 2. the application of a kind of phthalide derivative according to claim 1 in preventing and treating plant virus, plant pathogenic bacteria, plant pest mites, wherein, the compound shown in the general formula is selected from the compound shown in the following formula A kind of: 3-(3-methylbenzylidene)isobenzofuran-1(3H)-one (b); 3-(4-Butylbenzylidene)isobenzofuran-1(3H)-one (c); 3-(4-tert-Butylbenzylidene)isobenzofuran-1(3H)-one (d); 3-(4-Methoxybenzylidene)isobenzofuran-1(3H)-one (e); 3-(3-Chlorobenzylidene)isobenzofuran-1(3H)-one (f); 3-(4-Bromobenzylidene)isobenzofuran-1(3H)-one (g); 3-(2-Bromobenzylidene)isobenzofuran-1(3H)-one (h); 3-(thiophene-2-methylene)isobenzofuran-1(3H)-one (i); 3-benzylidene-5-methoxyisobenzofuran-1(3H)-one (j); 3-benzylidene-6-methylisobenzofuran-1(3H)-one (k); 3-benzylidene-5-chloroisobenzofuran-1(3H)-one (1); 3-Benzylidene-5-fluoroisobenzofuran-1(3H)-one (m).