阅读说明：本技术 杀菌剂抑霉唑离子液、及其制备方法和应用 (Bactericide imazalil ionic liquid, and preparation method and application thereof ) 是由 谭新球 张卓 陈岳 满益龙 曹永松 郑立敏 欧阳超 刘思珍 史晓斌 李成刚 张德 于 2020-07-29 设计创作，主要内容包括：本发明涉及农药领域,具体杀菌剂抑霉唑离子液、及其制备方法和应用。所述抑霉唑离子液是由抑霉唑与有机酸,更具体为脂肪酸、含苯环的酸的配体分别进行酸碱中和反应制备得到的离子液。实验结果表明,以辣椒炭疽病菌、稻瘟病菌、镰刀菌和辣椒疫霉菌为供试菌株,供试抑霉唑离子液对供试菌株表现出不同的增效抑菌活性,最高增效效果达到47.19％,具有广阔的应用前景。(The invention relates to the field of pesticides, in particular to imazalil ionic liquid as a bactericide, and a preparation method and application thereof. The imazalil ionic liquid is prepared by respectively carrying out acid-base neutralization reaction on the ligands of imazalil and organic acid, more specifically fatty acid and acid containing benzene ring. Experimental results show that the experimental imazalil ionic liquid shows different synergistic bacteriostatic activities on test strains by taking colletotrichum capsici, rice blast, fusarium and phytophthora capsici as the test strains, the highest synergistic effect reaches 47.19%, and the application prospect is wide.)

1. The ionic liquid of the bactericide imazalil is characterized by being prepared by respectively carrying out acid-base neutralization reaction on imazalil and organic acid, more specifically fatty acid, acid containing benzene ring and ligand.

2. The ionic liquid of claim 1, wherein said fatty acid is acetic acid, nonanoic acid, or oleic acid; the acid containing benzene ring is benzoic acid or salicylic acid.

3. The ionic liquid of claim 2, wherein the ionic liquid has the following structural formula:

4. the ionic liquid of claim 2, wherein said ligand is salicylic acid or oleic acid.

5. A bactericide comprising the ionic liquid according to any one of claims 1 to 4 as an active ingredient.

6. Use of an ionic liquid according to any one of claims 1 to 4 for controlling plant fungal diseases, wherein the fungal disease is caused by penicillium citrinum, colletotrichum, pyricularia oryzae, sclerotinia sclerotiorum, phytophthora capsici, fusarium, or ustilaginoidea virens, particularly pepper anthracnose, more preferably pepper colletotrichum.

7. A method of preparing an ionic liquid according to any one of claims 1 to 4, comprising the steps of:

weighing raw imazalil, adding organic alcohol, preferably methanol, and fully dissolving imazalil;

adding the ligand, and stirring for reaction at 50-80 ℃, preferably 60-70 ℃;

and removing organic alcohol from the reaction compound to obtain the ionic liquid.

8. The method of claim 7, wherein the ligand and imazalil are added equimolar.

9. The method of claim 7 or 8, wherein the reaction is followed to completion by thin layer chromatography.

10. The method of claim 7 or 8, wherein the removing methanol is removing methanol from the reaction mixture using a rotary evaporator.

Technical Field

The invention belongs to the field of chemical pesticides in plant protection, and particularly relates to an imazalil bactericide ionic liquid, and a preparation method and application thereof.

Background

The ionic liquid is salt which is completely composed of organic ions and has a melting point lower than 100 ℃, is generally in a molten state at room temperature, is different from the traditional molten salt, and is salt which is always in a liquid state and has relatively low viscosity at low temperature. The ionic liquid can optimize the physical and chemical properties of the matrix, such as melting point, solubility, surface tension and the like, by selecting different cation and anion pairing ions (different alkyl chain lengths, different ion types and the like). In recent years, scientists have taken active ingredients of medicines as the basis and matched with different types of paired anions/cations, so that the activity of the original medicine is ensured, the physicochemical properties of the defects of the original medicine are improved, and the influence of natural ionic liquid on the sustainable development of green chemistry causes enthusiasm attention of academics, thereby becoming one of the research hotspots.

In the field of agriculture, after the medicine is prepared into the ionic liquid, the permeability of the membrane can be improved, and the medicine release speed can be accelerated; in the field of agricultural chemistry, an appropriate ligand is selected to prepare an ionic liquid with hydrophobic property, so that the rain wash resistance is improved, the drift amount of active substances is reduced, the drug effect is enhanced, the usage amount of pesticides is reduced, and the movement of active ingredients in plants is promoted. Such as Rogers topic group [ Hough W L, Rogers R D. Ionic liquids the n and not from solvents to active pharmaceutical ingredients [ J ]. Bulletin of the chemical society of Japan,2007,80(12):2262-2269] by synthesizing ionic liquids, improving the hydrophobic, controlled release and conductive transport properties of the original drug to enhance the biological activity of the original drug; doherty et al [ Doherty K M, Kulpa Jr C F. Toxicity and antibacterial activity of imidazolium and pyridinium ionic liquids [ J ]. Green Chemistry,2005,7(4):185-189] synthesized imidazolium and pyridinium ionic liquids with increased antibacterial properties as the paired ionic alkyl chain grows; however, the current research on related ionic liquids is mainly limited to the improvement of the characteristics of the herbicide, such as stability, leaching property and the like, so that the control efficiency of the herbicide is improved, and the research on bactericides and other chemical pesticides is relatively low.

Imazalil belongs to broad-spectrum, systemic and substituted benzimidazole bactericides, has structural characteristics of typical cations, and influences cell membrane permeability, physiological functions and lipid anabolism. The mechanism of action is related to the nitrogen-containing heterocycle, and the nitrogen atom on the heterocycle can be bonded with the heme region (heme-iron active center) of cytochrome P450 sterol 14-alpha-demethylase (sterol 14 alpha-demethylase P450, Cyp51) in a coordination bond, thereby inhibiting the activity of the enzyme [ Ji H, Zhang W, Zhang M, et al Ji, H.et al, Structure-based de novo design, synthesis, and biological evaluation of non-azo inhibitors specific for lanosterol 14-demethylase of funngi. J.Med.chem.46,474-485[ J ]. Journal of Medicinal Chemistry,2003,46(4):474 485 ]. Ergosterol is an essential component of fungal cell membranes, Cyp51 is a key enzyme in the ergosterol biosynthetic pathway [ Yoshida Y, Aoyama Y, Noshiro M, et al, Sterol 14-demethylase P450(CYP51) providesa break through for the discission on the evolution of cytochrome P450 geneuperfamily [ J ]. Biochemical & Biophysic Research Communications,2000,273(3):799-804], inhibition of enzyme activity leading to a retardation of ergosterol synthesis, the absence of which leads to a disruption and a loss of function of the membrane structure, and finally to fungal death [ prunus leaf, Schulverns, Song Aihuan. antifungal CYP 153. is compared with the resistance Penicillium 51 gene sequence [ J. bacteriology report, 2003,22(1): 153: 156 ]. Imazalil belongs to dmi bactericides, and the ergosterol biosynthesis inhibitor dmi bactericides are positioned as low-to-medium-resistance risk medicaments by a bactericide resistance action committee (FRAC), but certain plant pathogenic bacteria can generate higher-level resistance or drug resistance on part of dmi bactericides due to unreasonable long-term use, and particularly, the penicillium citrinum has obvious drug resistance.

Currently, imazalil is mainly applied to control of penicillium citrinum, crop anthracnose pathogen and the like, and researches report that pathogenic fungi such as anthrax from various hosts generate drug resistance to various bactericides such as DMIs and the like, so that the problem of how to overcome or delay the generation and development of the drug resistance of pathogenic bacteria to the pathogenic bacteria so as to prolong the service life of the pathogenic bacteria is an important problem in the problem of the drug resistance of plant pathogenic bacteria. Meanwhile, the development speed of the novel bactericide is greatly limited due to long drug development period and high cost. The ionic liquid has the characteristics of easy synthesis, various varieties and stable physicochemical properties, and simultaneously has the characteristic of reducing environmental pollution by selecting different counter ions to improve the physicochemical properties of the pesticide on the premise of not influencing the biological activity of the pesticide, thereby providing a new way for the efficient use of the traditional broad-spectrum bactericide.

Disclosure of Invention

Based on the above, the organic acid, specifically the organic acid with different carbon chain lengths and containing benzene rings and heterocycles, is selected as the anion ligand, and imazalil is taken as the matrix (taken as the alkali), the imazalil ionic liquid is prepared through acid-base neutralization reaction, the chemical characterization of the related ionic liquid is researched, the biological activity and the synergistic condition of the ionic liquid prepared by different ligands are compared, and the change of the antibacterial spectrum is determined. The method is used for explaining the improvement of the sensitivity of fungi to the ionized medicines so as to delay the generation of drug resistance of the fungi, and simultaneously screening the optimal ligand so as to provide scientific basis for the preparation and the efficient utilization of the bactericide ionic liquid, prolong the service life of the high-efficiency but easily-generated drug imazalil, and simultaneously realize the reduction of the use amount of pesticides in agricultural production.

Therefore, the invention firstly provides the ionic liquid of the bactericide imazalil, which is prepared by respectively carrying out acid-base neutralization reaction on imazalil and organic acid, more specifically fatty acid, acid containing benzene ring or ligand with heterocyclic acid. The invention takes the fungicide imazalil as a matrix and takes organic acid with different carbon chain lengths and containing benzene rings and heterocycles as anion ligand to carry out chemical reaction, and the ionic liquid of which the product is the corresponding fungicide is obtained.

In a preferred embodiment, the fatty acid is acetic acid, nonanoic acid or oleic acid; the acid containing benzene ring is benzoic acid or salicylic acid. Wherein, the ionic liquid has the following structural formula:

in a more preferred embodiment, the ligand is salicylic acid.

The invention also provides a bactericide containing the ionic liquid as an active ingredient.

The invention also provides application of the ionic liquid in preventing and treating plant fungal diseases, wherein the fungal diseases are plant diseases caused by penicillium citrinum, colletotrichum, rice blast, sclerotinia sclerotiorum, phytophthora, fusarium or ustilaginoidea virens, particularly pepper anthracnose, and more preferably pepper colletotrichum.

Further, the present invention provides a method for preparing the above ionic liquid, which comprises the following steps:

weighing raw imazalil, adding organic alcohol, preferably methanol, and fully dissolving imazalil;

adding the ligand, and stirring for reaction at 50-80 ℃, preferably 60-70 ℃;

and removing organic alcohol from the reaction compound to obtain the ionic liquid.

Wherein in a particularly preferred embodiment, the ligand added and imazalil are added equimolar.

In one embodiment, the reaction is followed by thin layer chromatography to completion to determine when to perform the methanol removal operation.

In a more specific embodiment, the removing methanol is removing methanol from the reaction mixture using a rotary evaporator.

The imazalil ionic liquid is successfully prepared, and specifically comprises imazalil-acetic acid ionic liquid, imazalil-nonanoic acid ionic liquid, imazalil-oleic acid ionic liquid, imazalil-benzoic acid ionic liquid and imazalil-salicylic acid ionic liquid, the recovery yield is more than 89%, and the nuclear magnetic resonance hydrogen spectrum characteristics show that the chemical shifts of H near reaction sites participating in the reaction on imidazole rings in 6 ionic liquids are changed to different degrees, and the chemical shifts are 0.1ppm-0.4 ppm. The 6 ionic liquids have melting points lower than the self melting point (52.7 ℃) of imazalil, are liquid or wax at normal temperature and meet the preparation requirement of the ionic liquids. The inventor creatively takes imazalil as a parent (regarded as alkali) and obtains the imazalil ionic liquid by carrying out acid-base neutralization reaction with organic acid anion ligands with different carbon chain lengths and containing benzene rings and heterocycles. Experimental results show that the imazalil ionic liquid used as a test reagent and the pepper anthracnose, rice blast, fusarium and phytophthora capsici used as test strains show different synergistic bacteriostatic activities to the test strains. Wherein the imazalil-salicylic acid ionic liquid shows synergistic activity on colletotrichum capsici, rice blast and fusarium, and the maximum synergistic effect reaches 47.19 percent. In addition, imazalil is mainly used for preventing and controlling penicillium citrinum, is easy to generate resistance, is rarely used for pepper anthracnose pathogen, fusarium and the like, and shows that the imazalil can be used for preventing and controlling other bacteria in ionic liquid and has better effect according to the invention.

Drawings

FIG. 1 shows the NMR spectrum of imazalil.

FIG. 2 shows the NMR spectrum of imazalil-acetic acid ion liquid.

FIG. 3 shows the NMR spectrum of imazalil-nonanoic acid ionic liquid.

FIG. 4 shows the NMR spectrum of imazalil-oleic acid ionic liquid.

FIG. 5 shows the NMR spectrum of imazalil-benzoic acid ion liquid.

FIG. 6 shows the NMR spectrum of imazalil-salicylic acid ion liquid.

FIG. 7 determination of sensitivity of different Colletotrichum capsici to imazalil.

FIG. 8 sensitivity of colletotrichum gloeosporioides to imazalil and derivatives thereof.

FIG. 9 sensitivity of Pyricularia oryzae to imazalil and derivatives thereof.

FIG. 10 sensitivity of Fusarium to imazalil and its derivatives.

Detailed Description

In order to better understand the present invention, the following examples are further provided to illustrate the present invention, but not to limit the present invention.