Application of a nano-pesticide in inhibiting appressorium formation of Magnaporthe oryzae

Abstract

The application discloses application of a nano pesticide in inhibiting formation of appressorium of Pyricularia oryzae, wherein the nano pesticide is composed of copper ions and nano ZIF-8, and the copper ions are doped on the nano ZIF-8; wherein the mass ratio of the copper ions in the nano pesticide is 4.06 wt%. The nano pesticide has the advantages that not only can the formation of the appressorium of Pyricularia oryzae be inhibited, but also the production of active oxygen in mycelium can be induced, so that the morphological structure of the mycelium is caused to be oxidatively damaged; the nano pesticide mixed with conidia of Pyricularia oryzae is sprayed to inoculate rice leaves, the incidence of Pyricularia oryzae on the rice can be significantly reduced, and the prepared nano pesticide has no toxicity to the rice; Cu 2+ The copper ions doped into ZIF-8 and applied in inhibiting the formation of the appressorium of Pyricularia oryzae can not only improve the antibacterial activity of ZIF-8 on Pyricularia oryzae, but also reduce the pollution of traditional chemical pesticides to the environment.

Description

TECHNICAL FIELD

[0001] The application relates to application of a nano pesticide to inhibition of formation of appressorium of Magnaporthe oryzae, and belongs to the technical field of nano pesticides. BACKGROUND

[0002] Magnaporthe oryzae is one of the most destructive fungal diseases in rice production, widely distributed in rice fields all over the world, and mainly transmitted through air. It can infect the leaves, stems and panicles of rice, leading to a large area of yield reduction and seriously threatening global food security. When the disease is serious, it can cause a 10%-30% reduction in rice yield.

[0003] The pathogenic mechanism of Magnaporthe oryzae is complex and diverse. The conidia of Magnaporthe oryzae adhere to the plant surface by producing appressorium and break through the plant cell wall by mechanical force and chemical enzymatic action. After invading the cell, the pathogen rapidly reproduces to form mycelium and further spread, leading to the formation and expansion of lesions. This process is accompanied by a series of interactions between the pathogen and the host, including the secretion of effector proteins by the pathogen, the inhibition of host defense response and the generation of reactive oxygen species (ROS).

[0004] To control Magnaporthe oryzae, traditional methods rely on chemical fungicides and disease-resistant varieties. However, long-term use of chemical fungicides leads to environmental pollution and increased resistance of the pathogen, while the resistance of disease-resistant varieties often decreases over time. Therefore, finding new, more effective and environmentally friendly control methods has become the focus of current research. SUMMARY

[0005] The technical problem to be solved by the application is to provide application of a nano pesticide to inhibition of formation of appressorium of Magnaporthe oryzae, which not only inhibits the formation of appressorium of Magnaporthe oryzae, but also induces the generation of reactive oxygen species in mycelium, thereby causing oxidative damage to the morphological structure of mycelium. Spraying the nano pesticide mixed with conidia of Magnaporthe oryzae on rice leaves can significantly reduce the incidence of Magnaporthe oryzae on rice, and the prepared nano pesticide has no toxicity to rice.

[0006] The application is achieved by the following scheme: application of a nano pesticide to inhibition of formation of appressorium of Magnaporthe oryzae, wherein the nano pesticide is composed of copper ions and nano ZIF-8.

[0007] The copper ions are doped on the nano ZIF-8, and the mass fraction of copper ions in the nano pesticide is 4.06wt%.

[0008] The preparation method of the nano pesticide is as follows: during the self-assembly process of nano ZIF-8, copper ions are doped at the same time.

[0009] The preparation method of the nano pesticide is as follows:

[0010] Step one, dissolve zinc compound and copper compound and 2-methyl imidazole in water respectively;

[0011] Step two, first add zinc compound and copper compound into the aqueous solution and stir to mix evenly, then add the mixed solution into 2-methyl imidazole solution drop by drop.

[0012] The molar ratio of zinc ion: copper ion: 2-methyl imidazole is 2:1:120.

[0013] The zinc compound is zinc nitrate hexahydrate.

[0014] The copper compound is copper nitrate trihydrate.

[0015] The stirring time in step two is greater than or equal to 1 hour.

[0016] Application of nano-pesticide on the morphological structure of oxidative damage mycelium.

[0017] Application of nano-pesticide on the inhibition of Magnaporthe grisea conidial production.

[0018] The beneficial effects of the present application are:

[0019] 1. The present application can not only inhibit the formation of Magnaporthe grisea appressorium, but also induce the production of active oxygen in mycelium, thereby causing oxidative damage to the morphological structure of mycelium.

[0020] 2. The present application can significantly reduce the incidence of Magnaporthe grisea on rice by mixing the nano-pesticide with Magnaporthe grisea conidial spores and spraying it on rice leaves, and the prepared nano-pesticide has no toxicity to rice.

[0021] 3. The present application dopes Cu 2+ into ZIF-8 for inhibiting the formation of Magnaporthe grisea appressorium, which can not only improve the antibacterial activity of ZIF-8 on Magnaporthe grisea, but also reduce the environmental pollution caused by traditional chemical pesticides.

[0022] 4. The Cu(II) and ZIF-8 in the present application are mainly combined through physical adsorption and electrostatic attraction. When Cu-ZIF-8 is added to the culture medium, it will slowly decompose and release Cu(II) mixed with Zn(II) to inhibit the growth and sporulation of Magnaporthe grisea. At the same time, the interaction between nano-materials and Magnaporthe grisea mycelium will induce the production of excessive active oxygen (ROS) to damage the morphology and structure of mycelium.

[0023] 5、Rice blast appressorium formation generally requires a hydrophobic environment, and the addition of Cu-ZIF-8 will destroy the original hydrophobic environment, significantly inhibit the formation of rice blast appressorium, in addition, Cu-ZIF-8 itself and the metal ions released by it directly act on rice blast conidia, further inhibit the invasion and colonization of rice blast conidia on rice leaves;

[0024] 6、The application does not need complex and harsh conditions, has low preparation threshold and is easy to operate, can be completed at room temperature, and has low preparation cost;

[0025] 7、Cu-ZIF-8 has good safety for rice, is easily decomposed in the environment and can promote the growth of rice, and plays the role of preventing diseases and promoting crop growth. BRIEF DESCRIPTION OF DRAWINGS

[0026] Figure 1 a) is a scanning electron microscope image of ZIF-8 and Cu-ZIF-8.

[0027] Figure 1 b) is a transmission electron microscope image of ZIF-8 and Cu-ZIF-8.

[0028] Figure 1 c) is a particle size distribution graph of ZIF-8 and Cu-ZIF-8.

[0029] Figure 1 d) is XRD of ZIF-8 and Cu-ZIF-8.

[0030] Figure 1 e) is the FTIR graph of ZIF-8 and Cu-ZIF-8.

[0031] Figure 1 f) is a transmission electron microscope Mapping graph of ZIF-8 and Cu-ZIF-8.

[0032] Figure 2 a) is a direct graph of the influence of ZIF-8 and Cu-ZIF-8 at 0.1, 0.2 and 0.3 mg / mL on the growth of rice blast.

[0033] Figure 2 b) is a data statistical analysis graph of the influence of ZIF-8 and Cu-ZIF-8 at 0.1, 0.2 and 0.3 mg / mL on the growth of rice blast.

[0034] Figure 2 c) is a direct graph of the influence of ZIF-8 and Cu-ZIF-8 on the germination of rice blast conidia.

[0035] Figure 2d) Data statistical analysis of ZIF-8 and Cu-ZIF-8 on Magnaporthe grisea conidia germination.

[0036] Figure 2 e) Graphical representation of ZIF-8 and Cu-ZIF-8 at 0, 0.1, 0.05 and 0.025 mg / mL on Magnaporthe grisea conidia germination.

[0037] Figure 2 f) Data statistical analysis of ZIF-8 and Cu-ZIF-8 at 0, 0.1, 0.05 and 0.025 mg / mL on Magnaporthe grisea conidia germination.

[0038] Note: Different lower case letters indicate significant difference (P < 0.05).

[0039] Figure 3 a) Morphology of Magnaporthe grisea hyphae treated with different concentrations of ZIF-8 and Cu-ZIF-8.

[0040] Figure 3 b) Live and dead observation of Magnaporthe grisea treated with different concentrations of ZIF-8 and Cu-ZIF-8.

[0041] Figure 3 c) Reactive oxygen species production of Magnaporthe grisea treated with ZIF-8 and Cu-ZIF-8.

[0042] Figure 3 d) Different reactive oxygen species production of Magnaporthe grisea treated with Cu-ZIF-8 and hyphae.

[0043] Note: Lower case letters indicate significant difference (P < 0.05).

[0044] Figure 4 a) Invasive hyphae production of Magnaporthe grisea hyphae in rice leaf sheath cells treated with ZIF-8 and Cu-ZIF-8 (scale bar = 10 μm).

[0045] Figure 4 b) Disease development of Magnaporthe grisea on rice treated with different concentrations of ZIF-8 and Cu-ZIF-8 (0.1, 0.2 and 0.3 mg / mL).

[0046] Figure 4 c) Disease lesion area of Magnaporthe grisea on rice treated with different concentrations of ZIF-8 (0.1, 0.2 and 0.3 mg / mL).

[0047] Figure 4 d) Disease lesion area of Magnaporthe grisea on rice treated with different concentrations of Cu-ZIF-8 (0.1, 0.2 and 0.3 mg / mL).

[0048] Figure 5 a) The effect of ZIF-8 and Cu-ZIF-8 at different concentrations (0.1, 0.2 and 0.3 mg / mL) on the germination of rice seeds.

[0049] Figure 5 b) The effect of ZIF-8 and Cu-ZIF-8 at different concentrations (0.1, 0.2 and 0.3 mg / mL) on the germination rate of rice seeds.

[0050] Figure 5 c) The effect of ZIF-8 and Cu-ZIF-8 at different concentrations (0.1, 0.2 and 0.3 mg / mL) on the root length of rice seeds. DETAILED DESCRIPTION

[0051] The application will be further described below Figures 1-5 but the scope of the application is not limited to what is described.

[0052] For the sake of brevity, the full disclosure of each of the applications described below is incorporated herein by reference in its entirety, none of which is herein incorporated by reference for any other purpose or to the exclusion of other documents which can also be incorporated by reference. In the following description, numerous specific details are set forth to provide a thorough understanding of the present application. However, it will be apparent to one of ordinary skill in the art that the application can be practiced without such specific details. In other instances, well-known methods have not been described in detail in order to avoid obscuring the present application. It will be appreciated, that those who work in the art with the benefit of this disclosure will be able to devise their own implementations which, although perhaps not explicitly shown or described herein, embody the principles of the application and, thus, are within the spirit and scope of the application.

[0053] A method for preparing a nano-pesticide for inhibiting the formation of appressorium of Magnaporthe grisea, comprising the following steps:

[0054] Step one, 0.744g Zn(NO3)2.6H2O and 0.3g Cu(NO3)2.3H2O were dissolved in 10mL ddH2O, and ultrasonic treatment was performed for 5min at room temperature to make them completely dissolved.

[0055] Step two, 12.3g 2-methylimidazole was added to 90mL ddH2O, and ultrasonic treatment was performed for 10min at room temperature to make them completely dissolved.

[0056] Step three, the mixed solution of Zn(NO3)2.6H2O and 0.3g Cu(NO3)2.3H2O was added dropwise to the 2-methylimidazole solution, and continuous stirring was performed at 25℃ for 1h. The obtained precipitate was centrifuged for 10min (7830rpm / min), and washed with water for more than 3 times to completely remove the unreacted reactants, and the white precipitate was obtained by centrifugation, and washed with methanol for more than 3 times to completely remove the unreacted reactants, and the product dried at 65℃ overnight was Cu-ZIF-8.

[0057] The nano-pesticide prepared by the above method is subjected to experimental analysis.

[0058] Experimental analysis I, morphology and microstructure characterization, respectively using field emission scanning electron microscopy (SEM, Hitachi SU8010, Japan) and transmission electron microscopy (TEM, JEOL JEM-2100F, Japan).

[0059] X-ray diffraction patterns were collected using an X-ray diffractometer (XRD, Bruker D8 Focus, Germany) in the range of 5-35° with Cu Ka radiation at 40 kV and 40 mA. Fourier transform infrared spectroscopy (FT-IR, Nicolet IS10, Nicolet USA) was used to record the infrared spectra by KBr pellet method, with a scanning range of 4000 to 400 cm -1 , 32 scans and a resolution of 4 cm -1 . The particle size distribution of the nanomaterials was analyzed using Measurer 1.2 software. The experimental results are shown in Figure 1 .

[0060] From Figure 1 , it can be seen that ZIF-8 and Cu ion doped ZIF-8 (Cu-ZIF-8) are successfully prepared by one-pot method. XRD is used to analyze ZIF-8 and Cu-ZIF-8 materials. According to Figure 1 , it can be seen that the synthesized ZIF-8 sample crystal has 2θ of 7.3°, 10.1°, 12.5°, 14.8°, 16.5° and 17.7°, respectively, corresponding to (001), (002), (112), (022), (013) and (222) crystal faces, which are consistent with the main characteristic diffraction peaks of the ZIF-8 crystal structure matched with the standard card, indicating that the prepared ZIF-8 and metal ion doped ZIF-8 are both ZIF-8 crystals with high crystallinity, and it is proved that the use of "one-pot method" can realize the doping of metal ions, and the addition of Cu 2+ doping in the synthesis of ZIF-8 does not affect its crystal structure. SEM and TEM are used to characterize ZIF-8 and Cu-ZIF-8 nanocomposites, and the morphology of the nanoparticles is observed. As shown in Figure 1 , ZIF-8 shows uniform and regular dodecahedron structure with good crystallinity. Fourier transform infrared spectroscopy can determine the functional groups or chemical bonds present in the sample. The absorption peaks of ZIF-8 and Cu-ZIF-8 at 3135 cm -1 and 1580 cm -1 are related to the stretching vibration of C-H and C=N of imidazole, while the absorption peak at 412 cm -1The absorption peaks of the two materials are respectively related to the typical stretching vibration of Zn-N bond. The particle size distribution of the nanomaterials was analyzed by Measurer 1.2 software, and the results showed that the particle sizes of ZIF-8 and Cu-ZIF-8 were 294.55 ± 41.52 nm and 319 ± 30.91 nm, respectively. Meanwhile, the element distribution of the nanoparticles was characterized by TEM Mapping, and the results also showed that Cu 2+ ZIF-8 was successfully doped.

[0061] Experimental analysis two, the influence of ZIF-8 and Cu-ZIF-8 on the growth activity of Magnaporthe grisea: by measuring the growth diameter of the colony under different material concentrations, the biological activity of ZIF-8 and Cu-ZIF-8 on Magnaporthe grisea was tested. 5 mg of ZIF-8 and Cu-ZIF-8 powder was weighed and added to 5 mL of ddH2O, and 10 mg / mL of ZIF-8 and Cu-ZIF-8 stock solution was obtained after ultrasonic treatment for 10 min. 0, 300, 600, 900 μL of ZIF-8 and Cu-ZIF-8 stock solution was added to sterile molten CM solid medium, and the volume was made to 30 mL to prepare a series of CM medium with a concentration of 0, 0.1, 0.2 and 0.3 mg / mL of doped nanomaterials. After solidification, a puncher was used to take Magnaporthe grisea mycelium disc from the edge of the colony on the culture medium. After incubation at 28°C in a light incubator for 6 d, the inhibition rate (%) was measured. The influence of ZIF-8 and Cu-ZIF-8 on the appressorium formation of Magnaporthe grisea: 5 mg of Cu-ZIF-8 powder was weighed and added to 5 mL of ddH2O, and 10 mg / mL of Cu-ZIF-8 stock solution was prepared by ultrasonic treatment. The stock solution was diluted to 0, 200, 100, 50 μg / mL of 100 μL solution in a new EP tube. The spore solution of the strain Guy11 culture dish was taken, centrifuged at 5000 rpm for 5 min, and the supernatant was discarded and replaced with an appropriate amount of ddH2O. The spore solution was observed under a microscope using a hemocytometer, and the concentration was adjusted as appropriate. 100 μL of spore solution was added to the centrifuge tube containing 100 μL of material solution, and the mixture was mixed evenly to make the final concentration of the solution 0, 100, 50, 25 μg / mL. The above solution was dropped on a hydrophobic glass slide to induce germination, and after 8 h of treatment in the dark, the germination was observed, and the germination rate (%) and appressorium formation rate (%) were calculated; each treatment was repeated 3 times, and the experiment was repeated 3 times. The experimental results are shown in Figure 2 .

[0062] From Figure 2As can be seen, in this invention, to study the inhibitory activity of the synthesized nanomaterials against *Strombus oryzae*, a series of nanomaterials at concentrations (0.1, 0.2, and 0.3 mg / mL) were prepared, and their effects on the growth and appressorium formation of *Strombus oryzae* were investigated. The results showed that both ZIF-8 and Cu-ZIF-8 had good inhibitory effects on *Strombus oryzae*, and the inhibitory effect increased continuously with increasing concentration. Subsequently, based on the previous concentrations, the concentrations of the materials were further reduced (0.1, 0.05, and 0.025 mg / mL) to study the effect of the nanomaterials on the appressorium formation of *Strombus oryzae*. The results showed that the inhibitory effect decreased with decreasing nanomaterial concentration, but *Strombus oryzae* spores treated with Cu-ZIF-8 still exhibited a low appressorium formation rate; even at a concentration of 0.025 mg / mL, it could still significantly inhibit the germination of *Strombus oryzae* spores.

[0063] Experimental Analysis 3: Qualitative Staining Experiment of ZIF-8 and Cu-ZIF-8 on Live / Dead Cells of Rice Blast Fever: Rice blast fungus mycelial blocks were taken from the colony edge and placed in CM liquid medium. The blocks were cultured at 28℃ and 110 rpm for 36 h in a shaker, then filtered. 5 mg of Cu-ZIF-8 powder was weighed and added to 5 mL of ddH2O. The solution was dissolved by sonication to prepare 10 mg / mL ZIF-8 and Cu-ZIF-8 stock solutions. 0, 300, 600, and 900 μL of the ZIF-8 and Cu-ZIF-8 stock solutions were added to 50 mL centrifuge tubes, respectively. CM liquid medium was added to bring the volume to 30 mL, preparing a series of CM mediums with concentrations of 0, 0.1, 0.2, and 0.3 mg / mL of nanomaterial-doped materials. The previously filtered mycelia were evenly added to four centrifuge tubes. After culturing at 28℃ and 110 rpm for 8 h, mycelial balls were aspirated into 2 mL EP tubes. CM liquid culture medium was added to a final volume of 1 mL, and 30 μL of PI and SYTO9 dye were added. Staining was performed in the dark for 15-20 min. Mycelial slides were prepared and observed under a confocal fluorescence microscope to determine morphological changes and viability. Mycelia were treated with ZIF-8 and Cu-ZIF-8 at concentrations of 0.2 mg / mL. Reactive oxygen species (ROS) production was measured using a reactive oxygen species (ROS) detection kit. Simultaneously, EPR was used to measure hydroxyl radicals (·OH), superoxide radicals (·O2-), and singlet oxygen (·O2-) in Cu-ZIF-8 treated mycelia. 1 O2 generation. Experimental results are shown below. Figure 3 .

[0064] can be Figure 3 As can be seen, in order to study the effects of ZIF-8 and Cu-ZIF-8 nanomaterials on the mycelial morphology of rice blast fungus, confocal microscopy was used to observe the rice blast fungus treated with ZIF-8 and Cu-ZIF-8 nanomaterials. The results are as follows:Figure 4 As shown, the untreated Magnaporthe grisea hyphae grew normally, the hyphal surface was smooth, the hyphal outline was clear, and the morphology was full; however, the Magnaporthe grisea hyphae treated with ZIF-8 and Cu-ZIF-8 nanomaterials presented hyphal swelling, part of the hyphal cells appeared to be swollen, and the hyphal interior appeared to be broken and necrotic. Further, PI and SYTO9 were used for staining to observe the dead / alive condition of Magnaporthe grisea treated with ZIF-8 and Cu-ZIF-8 nanomaterials, and the observation results showed that Cu-ZIF-8 treatment caused more Magnaporthe grisea death, which was consistent with the previous experimental results. In addition, to study the antibacterial mechanism of Cu-ZIF-8, the active oxygen production of Magnaporthe grisea treated with Cu-ZIF-8 was observed, and the results showed that a large amount of ROS (green fluorescence) was produced in the cell after Magnaporthe grisea was treated with Cu-ZIF-8, and further EPR test was used to test the active oxygen production type of Magnaporthe grisea hyphae treated with Cu-ZIF-8, and it was found that Cu-ZIF-8 itself produced a small amount of active oxygen hydroxyl, and Magnaporthe grisea hyphae itself did not produce, but Cu-ZIF-8 could induce a large amount of hydroxyl radical (·OH), superoxide radical (·O2-) and singlet oxygen (O2) to be produced after treating Magnaporthe grisea hyphae, which indicated that Cu-ZIF-8 could induce Magnaporthe grisea to produce excessive accumulation of ROS, cause damage to the antioxidant system, make Magnaporthe grisea unable to grow normally, and further cause Magnaporthe grisea to die. 1 O2) production, indicating that Cu-ZIF-8 can induce Magnaporthe grisea to produce excessive accumulation of ROS, cause damage to the antioxidant system, make Magnaporthe grisea unable to grow normally, and further cause Magnaporthe grisea to die.

[0065] Experimental analysis four, rice (CO39) was cultivated for 21 days, and grew to the state of three leaves and one heart, the concentration of the tested strain of conidium was adjusted to 10 5

[0067] Figure 5 ​​​As can be seen, in order to verify the role of ZIF-8 and Cu-ZIF-8 in the pathogenic process of Magnaporthe oryzae, first, the spores of the Guy11 strain with a GFP label were mixed with ZIF-8 and Cu-ZIF-8 (0.1 mg / mL) for treatment, and then inoculated in rice leaf sheath cells, and the results showed that Cu-ZIF-8 could significantly inhibit the formation of Magnaporthe oryzae spores in the rice leaf sheath cells. Different concentrations of ZIF-8 and Cu-ZIF-8 were used to spray the rice seedlings, and the results of repeated experiments showed that ZIF-8 and Cu-ZIF-8 could significantly inhibit the occurrence of rice blast at a certain concentration, and the lesion area on the rice was significantly reduced, and Cu-ZIF-8 had a more excellent inhibitory effect than ZIF-8, and only 0.1 mg / mL could significantly inhibit the infection and pathogenesis of Magnaporthe oryzae on rice.

[0067] In experiment five, ZIF-8 and Cu-ZIF-8 were dissolved in pure water to prepare treatment solutions with concentrations of 100, 200 and 300 mg / L. 10 mL of treatment solution was added to each culture dish, and 50 sterilized whole rice seeds were placed in each treatment, and three replicates were set for each treatment. Pure water was used as a blank control. Cultivation was carried out in a light incubator (temperature 28°C, light / dark time: 12 hours / 12 hours, humidity 75%), and the treatment solution was replaced every day. After 5 days of incubation, 30 seeds were randomly selected from each treatment group, the germination rate was measured, and the root length and bud length were measured. The experimental results are shown in Figure 5 .

[0068] From Figure 5 As can be seen, after the rice was treated with ZIF-8 and Cu-ZIF-8 at concentrations of 0.1, 0.2 and 0.3 mg / mL for 5 days, the germination rate and root length of the rice seeds were counted. The results showed that compared with the control group, 0.1, 0.2 and 0.3 mg / mL of ZIF-8 and Cu-ZIF-8 had no adverse effect on the germination rate and root length of the rice seeds, and the germination rate of the seeds was more than 90%, and the average root length was more than 2 cm.

[0069] Although the technical solutions of the present application have been described and listed in detail, it should be understood that modifications or equivalent alternatives to the above embodiments can be made by those skilled in the art, and these modifications or improvements made without departing from the spirit of the present application are within the scope of the present application.

Claims

1. The use of a nano-pesticide in inhibiting the formation of appressorium of Magnaporthe grisea, characterized in that, The nano-pesticide is composed of a copper ion compound and nano-ZIF-8, and the preparation method of the nano-pesticide is as follows: In step one, a zinc ion compound, a copper ion compound and 2-methyl imidazole are respectively dissolved in water, wherein the molar ratio of the zinc ion compound, the copper ion compound and 2-methyl imidazole is 2:1:120; In step two, the zinc ion compound and the copper ion compound are first added into the aqueous solution and stirred to be uniformly mixed, and then the mixed solution is added dropwise into the 2-methyl imidazole solution.

2. The use of a nano-pesticide according to claim 1 for inhibiting the formation of appressoria of Magnaporthe grisea, characterized in that, The copper ion compound is doped on the nano-ZIF-8; wherein the mass ratio of the copper ion compound on the nano-pesticide is 4.06 wt%.

3. The use of a nano-pesticide according to claim 1 for inhibiting the formation of appressoria of Magnaporthe grisea, characterized in that, The preparation method of the nano-pesticide is as follows: the doping of the copper ion compound is simultaneously realized in the self-assembly process of the nano-ZIF-8.

4. The use of a nano-pesticide according to claim 1 for inhibiting the formation of appressoria of Magnaporthe grisea, characterized in that, The zinc ion compound is zinc nitrate hexahydrate.

5. The use of a nano-pesticide according to claim 1 for inhibiting the formation of appressoria of Magnaporthe grisea, characterized in that, The copper ion compound is copper nitrate trihydrate.

6. The use of a nano-pesticide according to claim 1 for inhibiting the formation of appressoria of Magnaporthe grisea, characterized in that, The stirring time in step two is greater than or equal to 1 hour.

7. The use of a nano-pesticide according to claim 1 for inhibiting the formation of appressoria of Magnaporthe grisea, characterized in that, The application of the nano-pesticide on the morphological structure of the oxidative damage mycelium.

8. The use of a nano-pesticide according to claim 1 for inhibiting the formation of appressoria of Magnaporthe grisea, characterized in that, The application of the nano-pesticide on the inhibition of the production of the rice blast spores.

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