Triazole compound for inhibiting plant growth as well as preparation method and application thereof
By developing a triazole compound containing amide groups, this compound inhibits plant growth by affecting the physiological process of auxin, solving the problems of existing triazole pesticides with high usage, high price and long half-life, and achieving the plant Effective inhibition of growth, especially the inhibition effect of roots is significant.
Patent Information
- Application Number
- CN202311029490.3
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2023-08-15
- Publication Date
- 2025-02-21
AI Technical Summary
Although existing triazole pesticides such as pareoxazole have high efficacy, their dosage, high price and long half-life are high, which can easily lead to residual problems and are highly toxic to fish, bees and other organisms, and in severe cases they endanger human health.
A triazole compound containing amide groups has been developed, which inhibits plant growth by affecting the physiological process of auxin. The preparation method includes reaction in dichlorosulfoxide, distillation under reduced pressure, washing of anhydrous dichloromethane, Condensation with 3-trifluoromethoxy-aniline and purified by column chromatography.
This compound can significantly inhibit plant growth, especially the inhibitory effect on the roots is more obvious, and no "rebound" was observed during the test cycle, providing a new design idea for triazole plant growth regulators.
Smart Images
Figure CN119490465A_ABST
Abstract
Description
Technical Field
[0001] The invention belongs to the technical field of pesticides and relates to a triazole compound for inhibiting plant growth and a preparation method and application thereof. Background Art
[0002] The strong systemicity of triazole pesticides makes them have the advantages of high efficacy and significant effects. Among them, paclobutrazol and linconazole are the most commonly used and widely applied. Studies have shown that both paclobutrazol and linconazole inhibit the synthesis of gibberellins by inhibiting the activity of kaurene isomerase, thereby causing dwarfing of plants. Although the effects of the two compounds are similar, the amount of paclobutrazol used is much greater than that of linconazole. On the one hand, although the activity of linconazole is 6 to 10 times that of paclobutrazol, due to the presence of a relatively active double bond in the structure of linconazole, the synthesis route is long and the synthesis conditions are harsh, so it is far inferior to paclobutrazol in terms of safety, production cost and stability; on the other hand, in addition to the function of plant growth regulation, paclobutrazol also has a bactericidal effect. Although paclobutrazol has outstanding advantages, it also has some disadvantages. For example, paclobutrazol is used in high concentrations (generally in the hundreds of ppm), is expensive (8,000 to 10,000 yuan / kg), and most importantly, has a long half-life (0.5 to 1 year), which easily leads to residual problems, making it highly toxic to fish, bees and other organisms, and in severe cases, harmful to human health. Therefore, it is very necessary to continue to develop new triazole small molecules with plant growth regulating activity.
[0003] During the research, we discovered a triazole compound containing an amide group, which has the effect of shortening plants and delaying growth. Its synthesis steps are short, the conditions are mild, and the raw materials are easily available. Further physiological test results show that the compound mainly inhibits plant growth by affecting the physiological process of auxin, and no "rebound" phenomenon was observed during the test period. Compared with existing triazole plant growth regulators, both the mechanism of action and the plant performance are completely different. These results will provide novel design ideas for the development of new triazole plant growth regulators. Summary of the invention
[0004] In view of this, one of the objects of the present invention is to provide a triazole compound for inhibiting plant growth; a second object of the present invention is to provide a method for preparing a triazole compound for inhibiting plant growth; a third object of the present invention is to provide an application of a triazole compound for inhibiting plant growth in inhibiting the growth of dicotyledons or monocotyledons.
[0005] In order to achieve the above object, the present invention provides the following technical solutions:
[0006] 1. A triazole compound for inhibiting plant growth, the structural formula of the triazole compound is as follows:
[0007]
[0008] 2. A method for preparing a triazole compound for inhibiting plant growth, the preparation method being as follows:
[0009] 1-(3-carboxyl-1,2,4-triazole)-yl-3,3-dimethyl-2-butanone is dispersed in dichlorothionyl, and the mixture is reacted at 80-83° C. for 0.5-1 h. After the reaction, the mixture is subjected to reduced pressure distillation and washed with anhydrous dichloromethane to obtain a 1-(3-acyl chloride-1,2,4-triazole)-yl-3,3-dimethyl-2-butanone intermediate. The intermediate is then placed in anhydrous dichloromethane, mixed and uniformly mixed, and then 3-trifluoromethoxy-aniline is added. The mixture is subjected to condensation reaction at 25-28° C. for 1.5-2 h. After the condensation reaction is completed, the triazole compound for inhibiting plant growth is obtained after purification.
[0010] Preferably, the molar volume ratio of the 1-(3-carboxyl-1,2,4-triazole)-yl-3,3-dimethyl-2-butanone and dichloride is 1:5-10, mmol:mL; in the condensation reaction, the molar volume ratio of the 1-(3-acyl chloride-1,2,4-triazole)-yl-3,3-dimethyl-2-butanone intermediate, anhydrous dichloromethane and 3-trifluoromethoxy-aniline is 1:10-25:1.5-2, mmol:mL:mmol.
[0011] Preferably, the purification method is column chromatography.
[0012] Preferably, the mobile phase in the column chromatography method is a mixed solvent formed by ethyl acetate, petroleum ether and methanol; the volume ratio of ethyl acetate, petroleum ether and methanol is 5-10:2-3:0.1-0.5.
[0013] 3. Use of the triazole compounds in inhibiting the growth of dicotyledonous plants or monocotyledonous plants.
[0014] Preferably, the dicotyledonous plant is Arabidopsis thaliana.
[0015] Preferably, the specific operation process of inhibiting the growth of dicotyledonous plants or monocotyledonous plants is any one of seed soaking, spraying, flushing or root irrigation.
[0016] The beneficial effects of the present invention are as follows: the present invention provides a triazole compound for inhibiting plant growth. The compound (ak) contains an amide group in its structure. Plant seeds are treated with different concentrations of ak in a seed soaking manner, and the root length and leaf length of the grown plant seedlings gradually shorten as the ak concentration increases. And at the same concentration, the root inhibition effect is significantly stronger than the leaf. Further monitoring of the free IAA concentration and the expression of Aux / IAA family members in the plant, the experimental results show that the compound ak can positively regulate the expression of IAA1, IAA2, IAA5, IAA11, and IAA19 in the Aux / IAA family, thereby promoting the accumulation of free IAA and inhibiting the growth of the plant.
[0017] The present invention also provides a method for preparing a triazole compound for inhibiting plant growth. The specific method is: firstly, 1-(3-carboxyl-1,2,4-triazole)-base-3,3-dimethyl-2-butanone is used to obtain a 1-(3-acyl chloride-1,2,4-triazole)-base-3,3-dimethyl-2-butanone intermediate, and then the intermediate is subjected to a condensation reaction with 3-trifluoromethoxy-aniline at room temperature. The preparation method is simple and easy to operate, the raw materials are easy to obtain, and it is suitable for large-scale production.
[0018] Other advantages, objectives and features of the present invention will be described in the following description to some extent, and to some extent, will be obvious to those skilled in the art based on the following examination and study, or can be taught from the practice of the present invention. The objectives and other advantages of the present invention can be realized and obtained through the following description. BRIEF DESCRIPTION OF THE DRAWINGS
[0019] In order to make the purpose, technical solutions and advantages of the present invention more clear, the present invention will be described in detail below in conjunction with the accompanying drawings, wherein:
[0020] Figure 1 is a structural diagram of the triazole compound (ak) for inhibiting plant growth prepared in Example 1;
[0021] Figure 2 The nuclear magnetic resonance hydrogen spectrum of the triazole compound (ak) for inhibiting plant growth prepared in Example 1 1 H NMR spectra;
[0022] Figure 3 The carbon NMR spectrum of the triazole compound (ak) for inhibiting plant growth prepared in Example 1 13 CNMR images;
[0023] Figure 4 is the nuclear magnetic resonance fluorine spectrum of the triazole compound (ak) for inhibiting plant growth prepared in Example 1;
[0024] Figure 5 is a high resolution mass spectrum (HRMS) of the triazole compound (ak) for inhibiting plant growth prepared in Example 1;
[0025] Figure 6 This is a comparison diagram of the effects of the triazole compound (ak) for inhibiting plant growth prepared in Example 1 on the roots and leaves of Arabidopsis seedlings at different concentrations;
[0026] Figure 7 This is a comparison chart of the effects of the triazole compound (ak) for inhibiting plant growth prepared in Example 1 and clean water (blank control) on the free IAA concentration in Arabidopsis plants;
[0027] Figure 8 This is a comparison of the effects of the triazole compound (ak) for inhibiting plant growth prepared in Example 1 and clean water (blank control) on the expression of Aux / IAA family members in Arabidopsis plants. DETAILED DESCRIPTION
[0028] The following describes the embodiments of the present invention by specific examples, and those skilled in the art can easily understand other advantages and effects of the present invention from the contents disclosed in this specification. The present invention can also be implemented or applied through other different specific embodiments, and the details in this specification can also be modified or changed in various ways based on different viewpoints and applications without departing from the spirit of the present invention. It should be noted that the illustrations provided in the following embodiments only illustrate the basic concept of the present invention in a schematic manner, and the following embodiments and features in the embodiments can be combined with each other without conflict.
[0029] Example 1
[0030] A triazole compound (ak) for inhibiting plant growth, the structural formula of the triazole compound is as follows Figure 1 The specific preparation method is as follows:
[0031] 371 mg (1.0 mmol) of 1-(3-carboxyl-1,2,4-triazole)-yl-3,3-dimethyl-2-butanone was dispersed in 10 mL of dithionyl chloride and reacted at 80°C for 0.5 h. After the reaction, the excess dithionyl chloride was removed by vacuum distillation, and then washed with anhydrous dichloromethane (3 times, 20 mL each time) to remove the residual dithionyl chloride and a small amount of hydrochloric acid and water to obtain 1-(3-acyl chloride-1,2,4-triazole)-yl-3,3-dimethyl-2-butanone. 3-dimethyl-2-butanone intermediate, take 1mmol of the intermediate and place it in 10mL of anhydrous dichloromethane, mix and mix well, add 265mg (1.5mmol) of 3-trifluoromethoxy-aniline, and condense at 25°C for 1.5h. After the condensation reaction, column chromatography (wherein the mobile phase is a mixed solvent formed by ethyl acetate, petroleum ether and methanol in a volume ratio of 5:3:0.5) is used for purification to obtain a white solid powder ak (yield is 78%).
[0032] Example 2
[0033] A triazole compound (ak) for inhibiting plant growth, the structural formula of the triazole compound is as follows Figure 1 The specific preparation method is as follows:
[0034] 371 mg (1.0 mmol) of 1-(3-carboxyl-1,2,4-triazole)-yl-3,3-dimethyl-2-butanone was dispersed in 5 mL of thionyl chloride and reacted at 83 °C for 1 h. After the reaction, the excess thionyl chloride was removed by vacuum distillation, and then washed with anhydrous dichloromethane (3 times, 20 mL each time) to remove the residual thionyl chloride and a small amount of hydrochloric acid and water to obtain 1-(3-acyl chloride-1,2,4-triazole)-yl-3,3- Dimethyl-2-butanone intermediate, take 1mmol of the intermediate and place it in 25mL of anhydrous dichloromethane, mix and mix well, add 353.33mg (2mmol) of 3-trifluoromethoxy-aniline, and condense at 28°C for 2h. After the condensation reaction, column chromatography (wherein the mobile phase is a mixed solvent formed by ethyl acetate, petroleum ether and methanol in a volume ratio of 10:2:0.1) is used for purification to obtain a white solid powder ak (yield is 78%).
[0035] The triazole compound (ak) for inhibiting plant growth prepared in Example 1 was subjected to melting point, nuclear magnetic resonance (hydrogen spectrum, carbon spectrum, fluorine spectrum) and high-resolution mass spectrometry tests for structural identification. The experimental results are as follows:
[0036] Melting point: 117-120°C;
[0037] 1H NMR (600MHz, CDCl3): δ9.35(s,1H),8.59(s,1H),7.75(s,1H),7.62(d,J=8.1Hz,1H),7.62(d,J=8.1Hz,1H),7.37 (t,J=8.2Hz,1H),7.37(t,J=8.2Hz,1H),7.02(d,J=8.1Hz,1H),7.02(d,J=8.1Hz,1H),5.42(s,2H),1.28(s,9H)(such as Figure 2 shown);
[0038] 13 C NMR (151 MHz, CDCl3): δ 205.49, 156.12, 155.73, 149.65, 145.67, 138.66, 130.20, 118.07, 116.85, 112.79, 54.47, 43.63, 26.13 (as Figure 3 shown);
[0039] 19 F NMR (565MHz, CDCl3): δ-57.73 (as Figure 4 shown);
[0040] HRMS(ESI): theoretical value [M+H] + C 16 H 17 F3N4O3: 371.1325; Measured value: 371.1324 (such as Figure 5 shown).
[0041] Compound ak 1 The H NMR spectrum shows that the aromaticity of 1,2,4-triazole makes the hydrogen chemical shift of 5-CH on the ring in the low field (δ=8.59ppm) and appears in the form of a single peak; the -CH2 connected to tert-butyl ketone is affected by the electron-withdrawing carbonyl group, and the chemical shift moves to 5.42ppm in the low field. The hydrogen chemical shift of the three -CH3 of tert-butyl group is 1.28ppm, and the chemical environment of these hydrogen atoms is almost the same. The peaks of 9 hydrogen atoms overlap and appear in the form of a single peak. The chemical shift of -NH hydrogen connected to the carbonyl group is in the lowest field (δ=9.35ppm). There are 4 hydrogens with different chemical shifts on the benzene ring. The aromatic hydrogen at the 2-position is not split because the ortho-position is substituted and appears in the form of a single peak (δ=7.75ppm). The hydrogen at the 4-position and 6-position are both double peaks with chemical shifts of 7.02 and 7.62 respectively. The hydrogen at the 5-position is split into a triplet due to the ortho-position hydrogen, and the chemical shift is 7.37.
[0042] 13C NMR spectrum analysis shows that: the ketone carbonyl carbon in the compound ak structure is in the low field (205.49ppm), and the carbonyl carbon in the amide has a chemical shift (156.12ppm); the methyl carbon in the tert-butyl structure has the highest chemical field (δ=26.13ppm), and the tert-carbon and the methylene carbon connected to the triazole are both adjacent to the carbonyl. Due to the steric effect of the methyl group in the tert-butyl group shielding electrons, the chemical shift of the tert-carbon (δ=43.63ppm) is lower than that of the methylene carbon (δ=54.47ppm). The carbon on the triazole ring has a lower chemical field than the carbon on the benzene ring due to the electron-withdrawing effect of the carbon group, so 155.73ppm and 149.65ppm are attributed to the carbon on the triazole ring, and the rest of the carbon is attributed to the carbon in the benzene ring.
[0043] HRMS spectrum shows: the molecular ion peak of compound ak is obtained by obtaining 1 H + The test source is formed. The test value has an accuracy of 4 decimal places, the theoretical value is 371.1325, and the test value is 371.1324, with a difference of 0.0001, which meets the evaluation requirements for determining the molecular weight of small molecule compounds by high-resolution mass spectrometry.
[0044] In summary, the above structural tests prove that the chemical structure of compound ak is Figure 1 The chemical structure is shown.
[0045] Performance Testing
[0046] 1. Investigate the effect of the triazole compound (ak) prepared in Example 1 for inhibiting plant growth on the roots and leaves of Arabidopsis seedlings
[0047] The biological activity test was conducted by the plate method. The specific method is as follows: Arabidopsis seeds were sown on culture media containing different ak concentrations (where the ak concentrations were set to 0 μmol / L, 0.1 μmol / L, 1 μmol / L, 5 μmol / L, 15 μmol / L, and 45 μmol / L), and the root length and leaf length under different concentration treatments were measured 15 days after sowing and recorded in Table 1.
[0048] Table 1 Effects of different concentrations of ak on the growth and development of Arabidopsis seedlings
[0049]
[0050]
[0051] As can be seen from Table 1, when the compound ak is in the range of 0 to 45 μmol / L, the root length decreases with the increase of ak concentration. Compared with Arabidopsis seedlings that have not been treated with ak, the inhibition rate of Arabidopsis seedling roots after treatment with 0.1 to 1 μmol / L ak is about 30%, and after treatment with 5 μmol / L ak, the inhibition rate of roots can be as high as about 80%. However, the inhibition rate of leaves at the same concentration is much lower than that of roots. For example, compared with Arabidopsis seedlings that have not been treated with ak, the change in leaf length after treatment with 0.1 μmol / L ak is not significant; when the concentration of ak is 1 μmol / L, the inhibition rate of leaves is 24% (this inhibition rate is only 80% of the root inhibition rate at the same concentration); when the concentration of ak is 5 μmol / L, the inhibition rate of leaves is 60%, which is lower than the inhibition level of roots at the same concentration. In order to more intuitively observe the effects of different concentrations of ak on the roots and leaves of Arabidopsis seedlings, the changes in the appearance and morphology of roots and leaves under different concentration treatments were also recorded. The experimental results are shown as follows. Figure 6 In conclusion, compound ak has a strong inhibitory effect on the whole plant (roots and leaves) of Arabidopsis thaliana, but the root response is more sensitive.
[0052] 2. Investigate the effect of the triazole compound (ak) prepared in Example 1 for inhibiting plant growth on the free IAA concentration and the expression of Aux / IAA family members in Arabidopsis plants
[0053] Arabidopsis seeds were spotted on a sterile solid culture medium (1 / 2MS) and cultured in an artificial climate chamber for 9 days. Some of the plants were cultured in an aqueous solution containing 30 μmol / L ak, and the other part of the plants were cultured in clean water (blank control). Samples were taken after 3 hours of culture, and the IAA content of the samples was determined (e.g. Figure 7 q-PCR technology was used to quantitatively detect IAA1, IAA2, IAA5, IAA11, and IAA19 in the Aux / IAA family (as shown in Figure 8 shown).
[0054] from Figure 7 It can be seen that compared with the water treatment, compound ak promoted the accumulation of free IAA in Arabidopsis plants. In order to further verify the reliability of the relationship between compound ak and IAA, we determined the expression levels of some of the earliest response genes of IAA (IAA1, IAA2, IAA5, IAA11, IAA19). The experimental results are as follows: Figure 8 As shown. Figure 8It can be seen that compound ak positively regulates the expression of IAA1, IAA2, IAA5, IAA11, and IAA19 in the Aux / IAA family. The above results show that compound ak may act directly or indirectly through IAA. Since IAA promotes plant growth at low concentrations and inhibits plant growth and development at high concentrations. However, when the minimum concentration of compound ak in this application was set to 0.1 μmol / L in this experiment, no growth promotion phenomenon was observed, which suggests that the concentration of compound ak may need to be set lower.
[0055] Similarly, the compound prepared in Example 2 was subjected to the same test as the compound in Example 1, and similar experimental results were obtained, thus once again proving that the preparation method in this application can be used to prepare a compound having Figure 1 The compound of the structural formula shown in the figure is used to treat plants with different concentrations of the compound, and the roots and leaves of the plants show different inhibitory effects. Further mechanism analysis shows that the compound in Example 2 also inhibits plant growth by affecting the physiological process of auxin.
[0056] In summary, the present invention provides a triazole compound for inhibiting plant growth. The compound (ak) contains an amide group in its structure. The compound can inhibit the growth of the entire plant, especially the root, which is the most sensitive. This inhibition mechanism is that the compound ak positively regulates the expression of IAA1, IAA2, IAA5, IAA11, and IAA19 in the Aux / IAA family, thereby promoting the accumulation of free IAA.
[0057] The present invention also provides a method for preparing a triazole compound for inhibiting plant growth. The method is simple and easy to operate, the raw materials are easy to obtain, and the method is suitable for large-scale production.
[0058] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the purpose and scope of the technical solution, which should be included in the scope of the claims of the present invention.
Claims
1. A triazole compound for inhibiting plant growth, characterized in that: The structural formula of the triazole compound is as follows:
2. The method for preparing a triazole compound for inhibiting plant growth according to claim 1, characterized in that: The preparation method is as follows: 1-(3-carboxyl-1,2,4-triazole)-yl-3,3-dimethyl-2-butanone is dispersed in dichlorothionyl, and the mixture is reacted at 80-83° C. for 0.5-1 h. After the reaction, the mixture is subjected to reduced pressure distillation and washed with anhydrous dichloromethane to obtain a 1-(3-acyl chloride-1,2,4-triazole)-yl-3,3-dimethyl-2-butanone intermediate. The intermediate is then placed in anhydrous dichloromethane, mixed and uniformly mixed, and then 3-trifluoromethoxy-aniline is added. The mixture is subjected to condensation reaction at 25-28° C. for 1.5-2 h. After the condensation reaction is completed, the triazole compound for inhibiting plant growth is obtained after purification.
3. The preparation method according to claim 2, characterized in that: The molar volume ratio of the 1-(3-carboxyl-1,2,4-triazole)-base-3,3-dimethyl-2-butanone and dithionyl chloride is 1:5-10, mmol:mL; in the condensation reaction, the molar volume ratio of the 1-(3-acyl chloride-1,2,4-triazole)-base-3,3-dimethyl-2-butanone intermediate, anhydrous dichloromethane and 3-trifluoromethoxy-aniline is 1:10-25:1.5-2, mmol:mL:mmol.
4. The preparation method according to claim 2, characterized in that: The purification method is column chromatography.
5. The preparation method according to claim 4, characterized in that: The mobile phase in the column chromatography method is a mixed solvent formed by ethyl acetate, petroleum ether and methanol; the volume ratio of ethyl acetate, petroleum ether and methanol is 5-10:2-3:0.1-0.
5.
6. Use of the triazole compound according to claim 1 in inhibiting the growth of dicotyledonous plants or monocotyledonous plants.
7. The use according to claim 6, characterized in that: The dicotyledonous plant is Arabidopsis thaliana.
8. The use according to claim 6, characterized in that: The specific operation process of inhibiting the growth of dicotyledonous plants or monocotyledonous plants is any one of seed soaking, spraying, flushing or root irrigation.