Use of a pyrimidine compound in the preparation of a preparation for preventing and treating plant root-knot nematodes

Abstract

The application discloses application of a pyrimidine compound in preparation of a preparation for preventing and treating plant root-knot nematodes. The pyrimidine compound is 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine. It is found for the first time that 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine has strong toxic effects on various plant root-knot nematodes such as Meloidogyne incognita, Meloidogyne javanica and M. enterolobii, and the half lethal concentration LC 50 of the pyrimidine compound on the nematodes is 10.91 μg / ml, 23.76 μg / ml and 12.96 μg / ml respectively in 24 hours, and the half lethal concentration LC 50 of the pyrimidine compound on the nematodes is 9.97 μg / ml, 26.16 μg / ml and 15.58 μg / ml respectively in 48 hours. In addition, the pyrimidine compound has no toxic activity on non-plant pathogenic nematodes such as Caenorhabditis elegans, has the characteristics of high efficiency and low toxicity, and can be used for development and research of root-knot nematode insecticides.

Description

TECHNICAL FIELD

[0001] The application belongs to the technical field of agricultural pest control, and particularly relates to application of a pyrimidine compound 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine in preparation of a preparation for controlling plant root-knot nematodes. BACKGROUND

[0002] Plant parasitic nematodes not only cause serious damage to the global ecological environment, but also bring huge economic losses to agricultural production. Among plant parasitic nematodes, root-knot nematodes (Meloidogyne spp.) that are parasitic to the roots of plants are the most widely distributed and most harmful parasitic nematodes, and are considered to be the "most destructive plant pathogen in the world". By infecting the roots of crops, root-knot nematodes affect the absorption of nutrients and water by crop roots, leading to the normal growth of plants being affected, and even causing the death of crops, greatly reducing the yield and quality of crops. After being infected by root-knot nematodes, crops also provide a channel for the infection of other crop diseases, causing greater harm to crops through complex infection and synergistic damage. Due to the particularity of infection and the concealment of disease, it is difficult to achieve an ideal control effect, and at present, root-knot nematodes have become a worldwide problem in the field of plant protection. Moreover, due to regional differences, the sensitivity of the same compound to root-knot nematodes may be different. However, traditional commercial chemical nematicides have been gradually abandoned due to problems such as high toxicity, carcinogenicity, and environmental pollution. Therefore, in-depth research on high-efficiency and low-toxicity nematicides, and the use of these compounds as lead compounds for new pesticide creation have become an important content of research on the prevention and control of root-knot nematode diseases. Meloidogyne spp. ) is a parasitic nematode that is most widely distributed and most harmful, and is considered to be the "most destructive plant pathogen in the world". By infecting the roots of crops, root-knot nematodes affect the absorption of nutrients and water by crop roots, leading to the normal growth of plants being affected, and even causing the death of crops, greatly reducing the yield and quality of crops. After being infected by root-knot nematodes, crops also provide a channel for the infection of other crop diseases, causing greater harm to crops through complex infection and synergistic damage. Due to the particularity of infection and the concealment of disease, it is difficult to achieve an ideal control effect, and at present, root-knot nematodes have become a worldwide problem in the field of plant protection. Moreover, due to regional differences, the sensitivity of the same compound to root-knot nematodes may be different. However, traditional commercial chemical nematicides have been gradually abandoned due to problems such as high toxicity, carcinogenicity, and environmental pollution. Therefore, in-depth research on high-efficiency and low-toxicity nematicides, and the use of these compounds as lead compounds for new pesticide creation have become an important content of research on the prevention and control of root-knot nematode diseases.

[0003] Trifluoromethyl, pyridine, pyrimidine, piperidine compounds are widely used in insecticidal compounds research and development, including triflumezopyrim, pirimicarb, dicyclanil, haloxyfop-methyl, flazasulfuron, Propyl 4-((2-((4,6-dimethoxypyrimidin-2-yl)oxy)benzyl)amino)[3,5-2H]benzoate, dimepiperate, piperophos, oxathiapiprolin, etc. Although trifluoromethyl, pyridine, pyrimidine, piperidine compounds have been reported as insecticides, the 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine of the present application has not been reported. d]pyrimidine(4-(4-((4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4- d ]pyrimidine) has been reported on its nematicidal activity and its use in the preparation of nematicidal preparations. Summary of the Invention

[0004] The present invention aims to provide the use of a pyrimidine compound, 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine, in the preparation of a plant root-knot nematode control formulation. The structural formula of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine is as follows: Figure 1 The compound was purchased from MCE with CASN 2380097-27-6 and serial number HY-Q16766. Its relative molecular mass is 376.34, and its mass-to-charge ratios m / z are 376.13 (100.0%), 377.13 (18.4%), and 377.12 (2.2%).

[0005] The present invention first discovered that 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine has a strong toxic effect on a variety of plant root-knot nematodes such as southern root-knot nematodes, Javan root-knot nematodes and elephant ear bean root-knot nematodes. The median lethal dose LC of the 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine is toxic to a variety of plant root-knot nematodes such as southern root-knot nematodes, Javan root-knot nematodes and elephant ear bean root-knot nematodes in 24 hours. 50 The concentrations were 10.91 μg / ml, 23.76 μg / ml and 12.96 μg / ml respectively; the median lethal dose LC50 of the southern root-knot nematode, the javanic root-knot nematode and the elephant-ear root-knot nematode was 2.37 μg / ml and 1.21 ... 50 The concentrations are 9.97μg / ml, 26.16μg / ml and 15.58μg / ml respectively. In addition, it has no toxic activity against non-plant pathogenic nematodes such as Caenorhabditis elegans, and is highly effective and low toxic, providing new ideas and approaches for the development of new root-knot nematode insecticides. BRIEF DESCRIPTION OF THE DRAWINGS

[0006] Figure 1 is the structural formula of the pyrimidine compound pyrimidine 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine of the present invention;

[0007] Figure 2 The calculation result of the toxicity determination of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine against southern root knotworm in Example 1;

[0008] Figure 3 Toxicity assay results of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine against Meloidogyne javanica in Example 1;

[0009] Figure 4 Toxicity assay results of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine against M. arenaria in Example 1;

[0010] Toxicity assay results of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine against M. incognita in Example 1. DETAILED DESCRIPTION

[0011] The present application will be further described in conjunction with the examples below, but in no way limits the present application, any transformation or improvement based on the teaching of the present application falls within the protection scope of the present application.

[0012] The present application provides the use of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine and / or its pesticidally acceptable salt in controlling plant root-knot nematodes.

[0013] The root-knot nematodes are Meloidogyne arenaria, M. incognita, M. javanica, M. arenaria or M. phaseolina.

[0014] The present application also provides a preparation for controlling plant root-knot nematodes, which takes 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine and / or its pesticidally acceptable salt as the main active ingredient or one of the active ingredients.

[0015] The preparation is aqueous solution, aqueous emulsion, wettable powder, soluble powder, soluble granule, water dispersible granule, suspension, oil suspension, tablet or effervescent granule.

[0016] The concentration of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine in the aqueous solution or aqueous emulsion is 1-50 μg / ml, and the solvent is aqueous solution of fatty alcohol or dimethyl sulfoxide with a concentration of ≤5%.

[0017] The fatty alcohol includes but is not limited to methanol, ethanol and propanol.

[0018] Example 1 Nematicidal activity detection of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine

[0019] 1. Sample preparation

[0020] Dissolve 0.5-1.5 mg of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine in DMSO and dilute with 4% DMSO to prepare the test solution. The dilution concentrations were 0.03 μg / ml, 0.05 μg / ml, 0.1 μg / ml, 0.3 μg / ml, 0.5 μg / ml, 1 μg / ml, 3 μg / ml, 5 μg / ml, 10 μg / ml, 30 μg / ml, and 50 μg / ml, respectively. The DMSO concentration in each solution was 0.4%. Water containing 0.4% DMSO served as a blank control.

[0021] 2. Preparation of Nematodes for Experimental Use

[0022] Tomato roots infected with southern root-knot nematodes were washed with water. Oocysts were picked with a dissecting needle and placed in a 0.4 M KCl solution. In a 5 ml centrifuge tube, the roots were washed and disinfected three times with 4 ml of sterile water, 1 ml of sterile penicillin-streptomycin solution, and 3 ml of 0.1% NaClO solution. The supernatant was removed by brief centrifugation at each step (8000 rpm for 30 seconds). The washed eggs were incubated in a 0.4 M ZnCl2 solution for 48 hours. Second-instar larvae were collected with a pipette into a 1.5 ml centrifuge tube and enriched by brief centrifugation at 5000 rpm for 30 seconds.

[0023] In addition, this experiment also prepared experimental nematodes such as Java root-knot nematode, elephant ear bean root-knot nematode and Caenorhabditis elegans according to conventional methods.

[0024] 3. Test methods

[0025] Transfer 100 μl of each test solution prepared in step 1 to a 20-well plate. Add 50-100 live nematodes prepared in step 2. Incubate at 25°C and calculate nematode mortality under a light microscope after 24, 48, and 72 hours. Identify nematode mortality by adding 1-5 drops of 5% NaCl solution to the treated wells. After 2 minutes, observe. Dead nematodes will be rigid and unresponsive to needle stimulation. Live nematodes will curl or twist.

[0026] The test solution without sample was used as the control, and the whole experiment was repeated three times. The average of the three times was taken to calculate the corrected mortality rate.

[0027] Mortality % = (number of dead nematodes / (number of dead nematodes + number of surviving nematodes)) × 100%

[0028] Corrected mortality % = sample mortality – control mortality

[0029] 4. Experimental results

[0030] 4.1 The toxicity of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine against southern root-knot nematodes is shown in Table 1 and Figure 2 As shown:

[0031] Table 1 Toxicity results of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine against southern root-knot nematode

[0032]

[0033] 4.2 The toxicity of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine against root-knot nematodes is shown in Tables 2 and Figure 3 shown.

[0034] Table 2 Toxicity of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine to root-knot nematodes of Javanese

[0035]

[0036] 4.3 The toxicity of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine against R. aurantii is shown in Tables 3 and Figure 4 As shown:

[0037] Table 3 Toxicity of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine to root knot nematodes of the ear bean

[0038]

[0039] 4.4 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine had no toxic activity against the non-plant pathogenic nematode Caenorhabditis elegans within 24 hours.

[0040] Result analysis: After determination ( Figures 2-4 ), the median lethal dose (LC50) of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine against southern root-knot nematodes, javanic root-knot nematodes and elephant ear bean root-knot nematodes in 24 hours 50 The concentrations were 10.91μg / ml, 23.76μg / ml and 12.96μg / ml respectively; the median lethal dose LC of the southern root-knot nematode, the javanic root-knot nematode and the elephant-ear root-knot nematode was 48 hours. 509.97 μg / ml, 26.16 μg / ml and 15.58 μg / ml, respectively. It is shown that 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine has strong toxic effect on plant nematodes such as Meloidogyne incognita, M. javanica and M. enterolobii, and has no toxic activity on non-plant pathogenic nematode C. elegans, and can be used for the prevention and treatment of plant nematodes.

[0041] Example 2

[0042] An insecticide for preventing and treating plant nematodes, which uses 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine as an active ingredient at a concentration of 50 μg / ml and uses 3% ethanol solution as a solvent.

[0043] Example 3

[0044] An insecticide for preventing and treating plant nematodes, which uses 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyrido[3,4-d]pyrimidine as an active ingredient at a concentration of 10 μg / ml and uses 2% methanol solution as a solvent.

[0045] The above examples are the preferred embodiments of the present application, but the embodiments of the present application are not limited to the above examples, and any changes, modifications, substitutions, combinations, simplifications made without departing from the spirit and principles of the present application are equivalent replacement modes, and are all included in the protection scope of the present application.

Claims

1. Use of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine and / or its pesticide-acceptable salts in the preparation of a formulation for controlling plant root-knot nematodes, wherein the concentration of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine is 1-50 μg / ml.

2. The application according to claim 1, characterized in that The root-knot nematode is the southern root-knot nematode, the Java root-knot nematode or the elephant-ear bean root-knot nematode.

3. A preparation for controlling plant root-knot nematodes, characterized in that: The preparation contains 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine and / or its pesticide-acceptable salt as active ingredients, the concentration of 4-(4-(4-(trifluoromethyl)pyrimidin-2-yl)oxy)piperidin-1-yl)pyridin[3,4-d]pyrimidine is 1-50 μg / ml, and the solvent is a fatty alcohol aqueous solution or a dimethyl sulfoxide aqueous solution with a concentration of ≤5%.

4. The preparation according to claim 3, characterized in that The preparation is an aqueous solution, an aqueous emulsion, a wettable powder, a soluble powder, a soluble granule, a water-dispersible granule, a suspension, an oil suspension, a tablet or an effervescent granule.

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