Application of higher aliphatic alcohol in aspects of bacteriostasis and sterilization

Abstract

The invention belongs to the technical field of crop disease and pest control, and discloses application of higher aliphatic alcohol in bacteriostasis and sterilization. The application specifically comprises application of higher fatty alcohol in preparation of a preparation for bacteriostasis and sterilization of fungi, application of higher fatty alcohol in preparation of a preparation for prevention and control of diseases caused by plant pathogenic fungi and application of higher fatty alcohol in preparation of a preparation for bacteriostasis and sterilization of oomycetes. The invention also discloses application of higher aliphatic alcohol in preparation of preparations for preventing and controlling diseases caused by plant pathogenic oomycetes. The higher fatty alcohol comprises one or a mixture of n-decanol and n-dodecanol. The higher fatty alcohol is applied to bacteriostasis and sterilization of crops, a green, environment-friendly and efficient prevention and control method is provided for diseases caused by plant pathogenic fungi and oomycetes, and the method is a new choice for crop disease prevention and control.

Description

Technical Field

[0001] The invention belongs to the technical field of crop disease and insect pest control, and specifically relates to the application of higher fatty alcohols in antibacterial and sterilization. Background Art

[0002] It is estimated that the annual global crop loss due to pests and diseases is equivalent to one-third of the potential harvest, of which diseases caused by plant pathogenic fungi and oomycetes account for more than 70% of the total plant diseases. At present, the prevention and control of plant diseases mainly rely on chemical agents, which have a short-term prevention and control effect, but the long-term single application of chemical agents has brought about resistance risks, pesticide residues and environmental pollution. Many problems have become increasingly prominent. Reducing the risk of pesticide-induced resistance and cross-resistance and creating efficient, environmentally safe and green new pesticides have become a new trend in global pesticide research and development.

[0003] At the same time, in production practice, different plant diseases caused by fungi and oomycetes are often confused and misjudged, resulting in the use of wrong disease control agents and the failure of disease control. The development of preparations that can inhibit both types of pathogens is of great value in agricultural production.

[0004] Studies have shown that fungal growth can be effectively inhibited by destroying the cell membrane structure of fungi. Many fungicides destroy the cell wall and cell membrane of fungi, leading to non-selective permeation of substances inside and outside the cells, and ultimately causing cell death.

[0005] Higher fatty alcohols are composed of a fatty chain consisting of multiple carbon atoms and a hydroxyl group at one end. They have a specific carbon chain length in their chemical structure, often have certain solubility and crystallinity, and have the characteristics of low toxicity, low irritation and non-flammability, which makes them a type of green and environmentally friendly chemicals. Summary of the invention

[0006] In order to effectively prevent and control the invasion of plant pathogenic fungi and oomycetes on crops, the present invention provides an application of a higher fatty alcohol in antibacterial and bacteriostatic effects.

[0007] To solve the above problems, the technical solution adopted by the present invention is as follows:

[0008] The present invention provides an application of a higher fatty alcohol in preparing a preparation for inhibiting and sterilizing fungi. The higher fatty alcohol comprises n-decanol, n-dodecanol or a mixture of the two.

[0009] Furthermore, the preparation can be used to inhibit and kill fungi and oomycetes by destroying the integrity of the cell wall, resulting in damage to the cell structure and outflow of endogenous substances, thereby causing growth inhibition, damage and death of fungal cells.

[0010] Furthermore, the preparation achieves its antibacterial and bactericidal application on fungi by destroying mitochondrial function and up-regulating the transcription of key genes in the oxidative phosphorylation pathway, thereby causing growth inhibition, damage and death of fungal cells.

[0011] In the present invention, the preparations include but are not limited to aqueous emulsions, emulsifiable concentrates, suspensions, wettable powders, dusts, granules, aqueous solutions, sprayable solutions, concentrated emulsions, aerosols, seed coatings, etc. All dosage forms that adopt the inventive concept of the present invention are applicable.

[0012] In another aspect, the present invention provides a use of a higher fatty alcohol in preparing a preparation for preventing and controlling diseases caused by plant pathogenic fungi, wherein the higher fatty alcohol comprises n-decanol, n-dodecanol or a mixture of the two.

[0013] Furthermore, the preparation achieves prevention and control of diseases caused by plant pathogenic fungi by destroying the integrity of the cell wall of the plant pathogenic fungi, resulting in damage to the cell structure, outflow of endogenous substances, and causing growth inhibition, damage and death of fungal cells.

[0014] Furthermore, the preparation achieves prevention and control of diseases caused by plant pathogenic fungi by disrupting mitochondrial function and upregulating transcription of key genes in the oxidative phosphorylation pathway, thereby causing growth inhibition, damage and death of fungal cells.

[0015] In the present invention, the preparations include but are not limited to aqueous emulsions, emulsifiable concentrates, suspensions, wettable powders, dusts, granules, aqueous solutions, sprayable solutions, concentrated emulsions, aerosols, seed coatings, etc. All dosage forms that adopt the inventive concept of the present invention are applicable.

[0016] In another aspect, the present invention provides a use of a higher fatty alcohol in preparing a preparation for inhibiting and sterilizing oomycetes, wherein the higher fatty alcohol comprises n-decanol, n-dodecanol or a mixture of the two.

[0017] In another aspect, the present invention provides a use of a higher fatty alcohol in preparing a preparation for preventing and controlling diseases caused by plant pathogenic oomycetes, wherein the higher fatty alcohol includes n-decanol, n-dodecanol, or a mixture of the two.

[0018] In the present invention, the preparations include but are not limited to aqueous emulsions, emulsifiable concentrates, suspensions, wettable powders, dusts, granules, aqueous solutions, sprayable solutions, concentrated emulsions, aerosols, seed coatings, etc. All dosage forms that adopt the inventive concept of the present invention are applicable.

[0019] Furthermore, the preparation is an aqueous emulsion, and the aqueous emulsion includes the higher fatty alcohol, an emulsifier, a thickener, and water.

[0020] Compared with the prior art, the present invention has the following effects:

[0021] The present invention utilizes advanced physiological and biochemical detection and transcriptome sequencing analysis technology to screen and determine that preparations containing higher fatty alcohols directly destroy the cell walls and mitochondria of fungal cells, resulting in damaged cell structures, outflow of endogenous substances and dysfunction of mitochondria, causing growth inhibition, damage and death of fungal cells, achieving the purpose of antibacterial and bactericidal effects, thereby achieving the prevention and control of diseases caused by plant pathogenic fungi; at the same time, higher fatty alcohols can also inhibit the growth of oomycetes, have antibacterial and bactericidal effects, thereby achieving the prevention and control of diseases caused by plant pathogenic oomycetes. The present invention applies higher fatty alcohols to the antibacterial and bactericidal effects of crops, provides a green, environmentally friendly and efficient prevention and control method for diseases caused by plant pathogenic fungi and oomycetes, and is a new choice for the prevention and control of crop diseases. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 This is a graph showing the mycelial growth of Saccharomyces cerevisiae under treatment with various higher fatty alcohols and emulsifiers;

[0023] Figure 2 This is the SEM image of Saccharomyces cerevisiae cells treated with emulsifier;

[0024] Figure 3 This is the SEM image of Saccharomyces cerevisiae cells treated with n-decanol;

[0025] Figure 4 This is the SEM image of Saccharomyces cerevisiae cells treated with n-dodecyl alcohol;

[0026] Figure 5 Single parameter histogram for flow cytometry of cell integrity;

[0027] Figure 6 Single parameter histogram for cell ROS flow cytometry detection;

[0028] Figure 7 This is the volcano map of differentially expressed genes in yeast cells after treatment with n-dodecyl alcohol and emulsifier for 5 minutes;

[0029] Figure 8 This is the volcano map of differentially expressed genes in yeast cells after treatment with n-dodecyl alcohol and emulsifier for 1 hour;

[0030] Fig. 9 This is the GO enrichment analysis of differentially expressed genes in yeast cells after treatment with n-dodecyl alcohol and emulsifier for 5 minutes;

[0031] Fig.10 This is the GO enrichment analysis of differentially expressed genes in yeast cells after treatment with n-dodecyl alcohol and emulsifier for 1 hour;

[0032] Fig.11This is the KEGG enrichment analysis diagram of differentially expressed genes in yeast cells after 1-dodecanol and emulsifier treatment for 5 minutes;

[0033] Fig.12 This is the KEGG enrichment analysis diagram of differentially expressed genes in yeast cells after treatment with n-dodecyl alcohol and emulsifier for 1 hour;

[0034] Fig.13 This is a graph showing the mycelial growth of Fusarium oxysporum FOC009 after 5 days of treatment with higher fatty alcohols;

[0035] Fig.14 This is a graph showing the mycelial growth of Fusarium graminearum under the treatment of different higher fatty alcohols after 5 days;

[0036] Fig.15 This is a graph showing the mycelial growth of Fusarium oxysporum under treatment with different higher fatty alcohols after 5 days;

[0037] Fig.16 This is a graph showing the mycelial growth of Colletotrichum gloeosporioides under treatment with different higher fatty alcohols after 5 days;

[0038] Fig.17 This is a graph showing the growth of litchi downy mildew hyphae under treatment with different higher fatty alcohols after 7 days. DETAILED DESCRIPTION

[0039] In order to make the technical scheme of the present invention more clearly understood by those skilled in the art, the following embodiments are listed for illustration. It should be pointed out that the following embodiments do not limit the protection scope of the present invention.

[0040] Unless otherwise specified, the raw materials, reagents or devices used in the following examples can be obtained from conventional commercial sources or by existing known methods.

[0041] Example 1

[0042] This embodiment provides an aqueous emulsion containing higher fatty alcohols, which is composed of the following components by weight percentage: 40% of n-dodecyl alcohol, 5% of n-hexadecanol, 4% of emulsifier, 3% of thickener, and the balance of water.

[0043] In this embodiment, the emulsifier is castor oil polyoxyethylene ether, and the thickener is carboxymethyl cellulose.

[0044] This embodiment also provides a method for preparing the aqueous emulsion, comprising the following steps:

[0045] Put n-dodecyl alcohol and n-hexadecyl alcohol into a container and heat to 55°C to melt. Then add an emulsifier into the container and process it through a high shear homogenizer at a speed of 4500 rpm for 10 minutes. Then add 55°C water into the container and process it through a high shear homogenizer at a speed of 10000 rpm for 20 minutes, then cool it to 40°C. Then add a thickener into the container and process it through a high shear homogenizer at a speed of 10000 rpm for 30 minutes to obtain the product.

[0046] Example 2

[0047] This embodiment provides an aqueous emulsion containing higher fatty alcohols, which is composed of the following components by weight: 40% fatty alcohol, 4% emulsifier, 3% thickener, and the balance water.

[0048] In this embodiment, the emulsifier is castor oil polyoxyethylene ether, and the thickener is carboxymethyl cellulose.

[0049] In this embodiment, the reagents and fatty alcohols correspond to each other as shown in Table 1:

[0050] Table 1 Selection of fatty alcohols in various reagents

[0051]

[0052] This embodiment also provides a method for preparing the aqueous emulsion, comprising the following steps:

[0053] Put fatty alcohol into a container and heat it to 75°C to melt, then add emulsifier into the container, process it through a high shear homogenizer at a speed of 4500 rpm for 10 minutes; then add water at 75°C into the container, process it through a high shear homogenizer at a speed of 10000 rpm for 20 minutes, and cool it down to 40°C; then add thickener into the container, process it through a high shear homogenizer, and rotate it at 10000 rpm for 30 minutes to obtain the product.

[0054] Example 3

[0055] This embodiment provides an aqueous emulsion, which is composed of the following components by weight percentage: 4% emulsifier, 3% thickener, and the balance water.

[0056] In this embodiment, the emulsifier is castor oil polyoxyethylene ether, and the thickener is carboxymethyl cellulose.

[0057] Put the emulsifier into a container and heat it to 55°C, and process it through a high shear homogenizer at a speed of 4500 rpm for 10 minutes. Then add 55°C water to the container, and process it through a high shear homogenizer at a speed of 10000 rpm for 20 minutes. Cool it to 40°C, and then add the thickener to the container, process it through a high shear homogenizer, and rotate it at 10000 rpm for 30 minutes to obtain the product.

[0058] Example 4

[0059] Effects of aqueous emulsions containing higher fatty alcohols on the growth of Saccharomyces cerevisiae and its mechanism

[0060] Saccharomyces cerevisiae is an extremely important yeast in human life. Its cells are mostly spherical or oval in shape. It mainly reproduces by budding. It can survive both under aerobic and anaerobic conditions and has strong adaptability. As a unicellular eukaryotic microorganism, Saccharomyces cerevisiae has a high degree of similarity in cell and organelle structure with animal and plant cells. The genes and biochemical pathways found in Saccharomyces cerevisiae are also highly conserved in higher organisms. It is often used as a model organism for studying eukaryotes and has significant genetic adaptability advantages.

[0061] It is quite common to use Saccharomyces cerevisiae as a model organism to study the antibacterial and bactericidal mechanisms of higher fatty alcohols against fungi and oomycetes.

[0062] (I) Effects of aqueous emulsions containing higher fatty alcohols on the growth of Saccharomyces cerevisiae

[0063] Test reagent: the aqueous emulsion prepared according to the present invention as listed in Table 2.

[0064] Table 2 Sample No.

[0065]

[0066] (2) Test fungi:

[0067] Saccharomyces cerevisiae W303-1A.

[0068] (3) Test methods

[0069] Prepare YPD basal medium: 10 g yeast extract, 20 g tryptone, 20 g glucose, 20 g agar powder, add water to 1000 mL. Sterilize at 115°C for 20 min.

[0070] The W303-1A strain stored at -80°C was streaked onto a YPD solid plate and cultured in a 30°C incubator for 3-5 days until a single colony appeared.

[0071] Pick a single colony, inoculate it into 5 mL YPD liquid medium, and culture it at 30°C, 220 rpm overnight, i.e. 12-16 h.

[0072] Pipette 1 mL of bacterial solution and add it to 50 mL of YPD liquid culture medium. Culture at 30°C and 220 rpm for 12-16 h.

[0073] 1 mL of sample was drawn, and the absorbance was measured at OD600. The concentration of each sample was adjusted by dilution with deionized water, and the value was OD600 = 1.0.

[0074] Add 100 μL of the bacterial solution diluted to OD600 = 1.0 to the first tube of the eight-tube PCR series, and then add 90 μL of sterile deionized water to the five tubes. Pipette 10 μL of bacterial solution from the first tube and add it to the second tube, pipette and mix, then pipette 10 μL from it and add it to the third tube, and repeat the above process until the eighth tube.

[0075] After thoroughly mixing on a vortex oscillator, use a multichannel pipette to draw up 4 μL of bacterial solution each time and spot it on the treated drug culture medium.

[0076] Place the culture medium upside down in a 30°C incubator and culture for 48 hours. When bacterial colonies grow, take photos and observe.

[0077] (4) Test results:

[0078] The growth of Saccharomyces cerevisiae in each treatment group is as follows Figure 1 As shown, it can be observed that: (a) Gradient dilution of Saccharomyces cerevisiae in the emulsifier control group can grow normally, appear pink, and have no obvious inhibitory effect. (b) Hexadecanol, hexadecanol, octadecyl alcohol, eicosanol, and docosanol treatment groups have no obvious inhibitory effect on Saccharomyces cerevisiae at a low concentration of 0.04% (V / V). (c) In the medium of octadecyl alcohol and tetradecyl alcohol, Saccharomyces cerevisiae showed a relatively slight inhibition phenomenon, and its color was mostly light pink or milky white. (d) Decacanol and dodecanol can significantly inhibit the growth of Saccharomyces cerevisiae.

[0079] It can be seen from the above test results that under low concentration treatment, hexadecanol, hexadecanol, octadecyl alcohol, eicosanol and docosanol do not produce significant physiological toxicity to Saccharomyces cerevisiae, while decanol and dodecanol can significantly inhibit the growth of Saccharomyces cerevisiae and have a strong toxic effect on Saccharomyces cerevisiae.

[0080] (II) Effect of aqueous emulsions containing higher fatty alcohols on the morphological growth of Saccharomyces cerevisiae

[0081] (1) Test reagents: Sample C, Sample D, and Sample J in Table 2;

[0082] (2) Test fungus: Saccharomyces cerevisiae W303-1A;

[0083] (3) Field emission scanning electron microscopy (FE-SEM) sample preparation

[0084] Prepare YPD basal medium: 10 g yeast extract, 20 g tryptone, 20 g glucose, 20 g agar powder, add water to 1000 mL. Sterilize at 115°C for 20 min.

[0085] Under sterile conditions, take an activated Saccharomyces cerevisiae W303-1A culture dish, pick a single colony, inoculate it into 5 mL YPD liquid culture medium, culture it at 30°C, 180 rpm for 12 h, then draw 1 mL of bacterial solution and add it to 100 mL YPD liquid culture medium, culture it at 30°C, 180 rpm on a shaker, and culture it for 12 h.

[0086] Samples C, D and J were added to the bacterial solution, each diluted 2500 times, i.e. 0.04% (V / V), and the culture was continued for 6 hours.

[0087] Use a pipette to draw 1 mL of bacterial solution into a 1.5 mL centrifuge tube, centrifuge at 2000 g, 4°C for 3 min, and discard the supernatant.

[0088] Fixation: Add 1.5 mL of 2.5% glutaraldehyde to the sample centrifuge tube and fix at 4°C for more than 24 hours.

[0089] Rinse: The fixed samples were rinsed with PBS (phosphate buffered saline) three times, 15 min each time, to remove glutaraldehyde on the sample surface.

[0090] Dehydration: Dehydrate the mycelium / spores using 50%, 70%, 80%, and 90% gradient ethanol, once for 15 min each time; then dehydrate with 100% ethanol three times for 10 min each time.

[0091] Replacement: Replacement was performed with 50%, 75%, and 90% gradient tert-butanol (tert-butanol was diluted with ethanol by volume), once each, each time for 10 min; then replacement was performed with 100% tert-butanol three times, each time for 10 min.

[0092] Finally, the samples were freeze-dried, gold-sprayed and the ultrastructural changes on the surface of fungal hyphae were observed using a scanning electron microscope.

[0093] (4) SEM image analysis

[0094] Field emission scanning electron microscopy (FE-SEM) was used to observe the changes in the morphology of Saccharomyces cerevisiae cells before and after treatment. It can be observed that: (a) Figure 2As shown in Figure 2, the surface of the Saccharomyces cerevisiae cells treated with the control emulsifier (sample J) was intact and in a regular ellipsoid or spherical shape. There was no obvious damage on the cell surface and the cells could bud and reproduce normally. (b) Figure 3 As shown in Figure 2, after 6 h of treatment with 0.04% (V / V) n-decanol (sample C), the cell morphology changed more seriously, the cell wall was damaged, the cells also contracted more rapidly, the entire surface of the Saccharomyces cerevisiae cells was obviously wrinkled, and most of them could not bud normally. (c) Figure 4 As shown, after being treated with 0.04% (V / V) n-dodecanol (sample D) for 6 h, most of the cell walls of Saccharomyces cerevisiae were damaged and concave.

[0095] From the above changes in the morphology of Saccharomyces cerevisiae cells, it can be seen that the treatment with high-grade fatty alcohol emulsion in water can damage the cell wall of Saccharomyces cerevisiae and seriously affect the growth of Saccharomyces cerevisiae.

[0096] (III) Using Saccharomyces cerevisiae as an alternative host to study the physiological and biochemical effects under stress

[0097] (1) Test reagents: Sample C, Sample D, Sample J in Table 2

[0098] (2) Test fungi:

[0099] Saccharomyces cerevisiae W303-1A

[0100] (3) Preparation of Saccharomyces cerevisiae suspension

[0101] The cryopreserved Saccharomyces cerevisiae was inoculated on YPD solid medium and cultured at 30°C for 48 h.

[0102] Pick a single colony of Saccharomyces cerevisiae, inoculate it into 5 mL of YPD liquid culture medium, and culture it at 30°C, 180 rpm overnight, i.e., about 12 h.

[0103] Pipette 1 mL of bacterial solution and add it to 100 mL of YPD liquid culture medium. Incubate at 30°C and 180 rpm in a shaker for 13-16 h until the Saccharomyces cerevisiae cells reach the logarithmic growth phase and the OD600 value is about 0.8.

[0104] The cells were collected in the culture system, and centrifuged at 4°C, 4000 g for 15 min to collect the Saccharomyces cerevisiae cells.

[0105] Rinse twice with 0.01M PBS, collect the cells, and resuspend them in the same volume of PBS as before. Prepare bacterial suspension with the bacterial solution that matches the culture time.

[0106] 1 mL of Saccharomyces cerevisiae suspension cultured to the logarithmic phase was added to the liquid culture medium containing 0.04% (V / V) of n-decanol and n-dodecanol, respectively, so that the total volume of the culture system was 30 mL, and cultured at 30°C, 180 rpm in the dark, and the culture medium system with the same concentration of emulsifier was used as a control. Saccharomyces cerevisiae cells were collected for analysis 6 hours after the start of culture.

[0107] (4) Detection of cell permeability

[0108] (a) Cell permeability detection method:

[0109] Add n-decanol, n-dodecanol and emulsifier to the suspension of Saccharomyces cerevisiae, i.e., n-decanol, n-dodecanol and emulsifier at a concentration of 0.04% (V / V), and culture at 30°C for 0 min, 5 min, 10 min, and 15 min. After reaching the specified time, centrifuge at 4°C, 4000g for 5 min, and collect the supernatant of each bacterial solution. Use a multifunctional microplate reader to measure the absorbance of the supernatant at 260nm and 280nm. Calculate the nucleic acid and protein concentrations respectively according to the following formulas.

[0110] DNA concentration (μg / mL) = 62.9 × A260 - 36.0 × A280;

[0111] Protein concentration (μg / mL) = 1552×A280-757.3×A260.

[0112] (b) Cell permeability test results:

[0113] When the cell wall of eukaryotic cells is damaged, the macromolecular contents in the cells, such as nucleic acids and proteins, will leak into the culture medium. The absorbance values ​​at 260nm and 280nm are widely used as markers for detecting the leakage of nucleic acids and proteins from the cell to the outside of the cell. As shown in Tables 3 and 4, there was no significant change in the extracellular DNA and protein of yeast cells after treatment with emulsifiers, indicating that the emulsifier as a control group had no significant effect on the permeability of the cell wall and cell membrane of yeast cells under this experimental treatment, while the leakage of DNA and protein in yeast cells increased significantly after treatment with n-decanol and n-dodecanol, which also indicated that the permeability of the cell wall and cell membrane of yeast cells increased after treatment. In addition, the concentration of extracellular leakage substances was positively correlated with the treatment duration.

[0114] Table 3 Extracellular DNA concentration of Saccharomyces cerevisiae under each treatment (μg / mL)

[0115]

[0116] Table 4 Extracellular protein concentration of Saccharomyces cerevisiae under each treatment (μg / mL)

[0117]

[0118] (4) Detection of cell integrity

[0119] (a) Cell integrity detection method:

[0120] The cells were collected in the above culture system, and centrifuged at 4°C, 4000 g for 10 min to collect the Saccharomyces cerevisiae cells.

[0121] After collecting the yeast cells, they were rinsed twice with PBS, centrifuged at 4000g for 5 min after each rinse, and the supernatant was discarded.

[0122] Then add an equal volume of PBS to make a bacterial suspension and dispense it into 1 mL to 1.5 mL EP tubes.

[0123] The hyphae were stained with 100 μL of propidium iodide (PI) solution (20 μg / mL).

[0124] The plates were incubated at 37°C in the dark for 15 min and then rinsed three times with PBS solution.

[0125] After adding 1 mL of PBS and mixing, aspirate 500 μL into a flow cytometer tube for flow cytometry detection. Count 10,000 cells. Repeat the experiment 3 times.

[0126] (b) Cell integrity test results:

[0127] Propidium iodide (PI), a nucleic acid dye, is repellent to cells with intact cell membrane structures and cannot penetrate them. However, when the cell membrane permeability changes or the cell membrane structure is damaged, PI dye can smoothly enter these cells and tightly bind to the DNA or RNA in the cells, emitting red fluorescence.

[0128] The single parameter histograms of flow cytometry under each sample treatment are as follows Figure 5 As shown in Table 5, under the treatment of emulsifier, the barrier function of the cell wall and cell membrane of Saccharomyces cerevisiae cells is normal, and PI can hardly enter the cells for staining; however, after treatment with n-decanol and n-dodecyl alcohol, the barrier function is destroyed, and the ratio of stained cells increases sharply, to 97.59% and 91.30%, respectively. The more dyes enter, the greater the fluorescence intensity. As can be seen from Table 5, the fluorescence intensity of Saccharomyces cerevisiae treated with n-decanol and n-dodecyl alcohol is much greater than that of Saccharomyces cerevisiae treated with emulsifier, which indicates that n-decanol and n-dodecyl alcohol can significantly affect the cell membrane permeability of Saccharomyces cerevisiae.

[0129] Table 5 Average fluorescence intensity of PI in Saccharomyces cerevisiae cells under various treatments

[0130] Reagents 1 2 3 Emulsifier 14211 13640 11980 n-Decanol 23037 22915 21464 n-Dodecanol 19979 19949 21525

[0131] (5) Detection of cellular reactive oxygen species (ROS)

[0132] (a) Cellular reactive oxygen species detection method:

[0133] After culturing for 0, 3, and 6 hours, cells were collected in the above culture system, centrifuged at 4°C, 4000g for 10 minutes, and Saccharomyces cerevisiae cells were collected. After collecting the yeast cells, rinse them twice with PBS, centrifuge them at 4000g for 5 minutes after each rinse, and discard the supernatant. Then add an equal volume of PBS to make a bacterial suspension, and dispense it into 1mL to 1.5mL EP tubes. Add 10μL of the active oxygen positive control reagent Rosup, mix well, and incubate at 37°C for 20 minutes. Add 200μL of the active oxygen dye DCFH-DA to the other treatment group, mix well, and incubate at 37°C in the dark for 20 minutes. Rinse with PBS 3 times.

[0134] After adding 1 mL of PBS and mixing, aspirate 500 μL into a flow cytometer and detect using a flow cytometer. Count 10,000 cells. Repeat the experiment 3 times.

[0135] (b) Cellular reactive oxygen species detection

[0136] Reactive oxygen is a key intermediate in aerobic metabolism. When cells face external stress, the level of ROS rises sharply, which is an important feature of cell stress response. Appropriate ROS can regulate cell growth, proliferation and differentiation, but excessive ROS seriously threatens cell structure, especially by oxidizing unsaturated fatty acids in cell membranes, inducing plasma membrane peroxidation, and damaging cell membrane integrity and function. At the same time, excessive ROS can also destroy mitochondrial function, weaken mitochondrial membrane potential, and affect energy metabolism.

[0137] The experiment used cells without dye staining as negative control and cells stained with Rosup as positive control. The single parameter histograms of each sample detected by flow cytometry were as follows: Figure 6 As shown in Table 6, the number of positive cells stained under emulsifier treatment was low, at 1.86%, while the number of positive cells stained under n-decanol and n-dodecanol treatment was large, at 11.92% and 11.8%, respectively. From the changes in fluorescence intensity under each treatment in Table 6, it was found that the fluorescence intensity of Saccharomyces cerevisiae under n-decanol and n-dodecanol treatment was significantly different from that of the emulsifier control.

[0138] Table 6 Average fluorescence intensity of ROS in Saccharomyces cerevisiae cells under each treatment

[0139] Reagents 1 2 3 Emulsifier 2212 2252 2050 n-Decanol 7669 7353 7172 n-Dodecanol 6962 7518 6772

[0140] (6) Detection of MDA (malondialdehyde) content in Saccharomyces cerevisiae

[0141] (a) Detection of MDA content in Saccharomyces cerevisiae

[0142] Collect cells in the above culture system, centrifuge at 4°C, 4000g for 10 minutes, and collect Saccharomyces cerevisiae cells. After collecting yeast cells, wash them twice with PBS, centrifuge at 4000g for 5 minutes after each wash. Add an equal volume of PBS and mix well, and ultrasonically disrupt (200W power, ultrasonic for 3s, interval of 10s, repeat 30 times). Centrifuge at 4000g for 5 minutes to collect the supernatant, and perform the assay according to the instructions of the kit.

[0143] (b) Results of MDA content detection in Saccharomyces cerevisiae

[0144] When the level of intracellular reactive oxygen species (ROS) increases abnormally, strong oxidative molecules will invade the cell membrane and protein structure, triggering the peroxidation of membrane lipids. Malondialdehyde (MDA), as a key byproduct of membrane lipid peroxidation, is widely regarded as a key indicator for assessing the degree of membrane lipid peroxidation. MDA can not only directly reflect the peroxidation level of cell membrane lipids, but also has the ability to interact with multiple components in the cell, further amplifying the destructive effect of ROS and exacerbating cell membrane damage.

[0145] As can be seen from Table 7, as the stress time increases, the intracellular MDA content also increases. Compared with the emulsifier control group, the intracellular MDA content of Saccharomyces cerevisiae treated with n-decanol and n-dodecyl alcohol increased significantly, but the intracellular MDA content of Saccharomyces cerevisiae treated with n-decanol was higher and the degree of membrane lipid oxidation was higher, which indicates that n-decanol has a stronger toxic effect on Saccharomyces cerevisiae.

[0146] Table 7 MDA content in Saccharomyces cerevisiae cells under different treatments (nmol / mL)

[0147]

[0148] (7) Detection of cell mitochondrial membrane potential

[0149] (a) Cell mitochondrial membrane potential detection method

[0150] Collect cells in the above culture system, centrifuge at 4000g for 10min at 4℃ to collect Saccharomyces cerevisiae cells. After collecting yeast cells, wash twice with PBS, centrifuge at 4000g for 5min after each wash. Add an equal volume of PBS to make a bacterial suspension, and dispense into 1mL to 2mL EP tubes. First, add 500μL of CCCP dye diluted to 10μM to the sample of the positive control group, and incubate at 37℃ for 25min. Add 500μL JC-1 staining solution to the pretreated cells, mix well, and incubate at 37℃ in the dark for 15min. After adding 500μL JC-1 staining buffer, centrifuge at 4000g for 5min, and discard the supernatant. After adding 1mL JC-1 staining buffer, centrifuge at 4000g for 5min, and discard the supernatant. Repeat the rinse with JC-1 staining buffer twice. After discarding the supernatant, add 1mL JC-1 staining buffer, mix well, and then detect. 10 μL of yeast suspension was pipetted onto a glass slide, and the prepared samples were observed morphologically using a fluorescence microscope and photographed for record.

[0151] (b) Cell mitochondrial membrane potential detection results

[0152] Mitochondria, as the key site for the synthesis of adenosine triphosphate (ATP) in cells, are crucial for the stable maintenance of their membrane potential for energy conversion. The reduction of mitochondrial membrane potential is widely recognized as one of the early biological markers of apoptosis.

[0153] The experiment used emulsifier-treated cells as negative control, and CCCP-stained cells as positive control. It can be found from Tables 8A, 8B, and 8C that compared with the emulsifier-treated group, the green fluorescence of the n-decanol and n-dodecanol treatment groups was significantly improved, showing significant differences. RFI also further demonstrated that n-decanol and n-dodecanol were significantly toxic to Saccharomyces cerevisiae, which could lead to a sharp drop in mitochondrial transmembrane potential, thereby causing a rapid weakening of mitochondrial activity.

[0154] Table 8A Fluorescence intensity (red light) of mitochondrial transmembrane potential of Saccharomyces cerevisiae cells under different treatments

[0155] 1 2 3 Emulsifier 116.8 113.9 127.5 n-Decanol 42.5 48.0 42.6 n-Dodecanol 34.2 35.4 32.5

[0156] Table 8B Fluorescence intensity (green light) of mitochondrial transmembrane potential of Saccharomyces cerevisiae cells under different treatments

[0157] 1 2 3 Emulsifier 4.04 3.87 4.11 n-Decanol 39.22 44.58 38.44 n-Dodecanol 36.32 39.50 37.36

[0158] Table 8C Fluorescence intensity (RFI) of mitochondrial transmembrane potential of Saccharomyces cerevisiae cells under different treatments

[0159] 1 2 3 Emulsifier 28.94 29.56 31.03 n-Decanol 1.08 1.08 1.11 n-Dodecanol 0.94 0.90 0.86

[0160] (IV) Transcriptome analysis of the effects of aqueous emulsions containing higher fatty alcohols on the growth of Saccharomyces cerevisiae

[0161] (1) Test reagents: Sample D and Sample J in Table 2

[0162] (2) Test fungi:

[0163] Saccharomyces cerevisiae W303-1A

[0164] (3) Preparation of transcriptome samples

[0165] Each bacterial solution (OD600 is about 0.8) was treated with 0.04% (V / V) n-dodecanol (sample D) at 30°C for 5 min and 1 h, and the emulsifier (sample J) treatment group was used as the control group. Three biological replicates were set for both the control group and the experimental group. Wuhan Maiwei Metabolic Biotechnology Co., Ltd. was commissioned to perform RNA extraction and RNA-Seq sequencing.

[0166] (4) Transcriptome analysis and results

[0167] (a) Screening of differentially expressed genes

[0168] Based on the sequencing results, DESeq2 was used to screen the differentially expressed genes. The screening conditions were |log2Fold Change|≥1 and FDR<0.05 to obtain differentially expressed genes.

[0169] Such as the volcano plot of differentially expressed genes ( Figure 7 , Figure 8 ) showed that compared with the control group, 4333 differentially expressed genes were generated after 5 min of treatment of Saccharomyces cerevisiae with n-dodecyl alcohol, including 877 up-regulated genes and 620 down-regulated genes;

[0170] Compared with the control group, 4115 differentially expressed genes were generated in Saccharomyces cerevisiae after treatment with n-dodecanol for 1 h, including 1072 up-regulated genes and 650 down-regulated genes.

[0171] (b) GO enrichment analysis of differentially expressed genes

[0172] After 5 min of treatment with n-dodecyl alcohol, the differentially expressed genes were mainly distributed in 15 biological processes, 15 cell components and 15 molecular functions. Fig. 9These differentially expressed genes were significantly enriched, and the 20 most significantly enriched GO-Terms were displayed in the form of a scatter plot. The results showed that GO-Term included mitochondrial ribosome, organellar ribosome, mitochondrial matrix, mitochondrial translation, mitochondrial protein complex, mitochondrial small ribosomal subunit, organellar small ribosomal subunit, organellar inner membrane, mitochondrial inner membrane, mitochondrial gene expression, etc.

[0173] After Saccharomyces cerevisiae was treated with n-dodecyl alcohol for 1 h, the differentially expressed genes were mainly distributed in 15 biological processes, 2 cellular components and 15 molecular functions. Fig.10 These differentially expressed genes were significantly enriched, and the 20 most significantly enriched GO-Terms were displayed in the form of a scatter plot. The results showed that the differentially expressed genes were mainly mitochondrial protein-containing complex, mitochondrial inner membrane, organelle inner membrane, mitochondrial membrane, mitochondrial matrix, mitochondrial ribosome, etc.

[0174] The results of GO enrichment analysis showed that the stress of n-dodecyl alcohol for 5 minutes and 1 hour affected the mitochondria and organelles of Saccharomyces cerevisiae. This indicates that n-dodecyl alcohol treatment may cause greater damage to the mitochondria or other organelles of Saccharomyces cerevisiae.

[0175] (c) KEGG enrichment analysis of differentially expressed genes

[0176] Fig.11After KEGG annotation, it was found that the differentially expressed genes in Saccharomyces cerevisiae after 5 minutes of n-dodecyl alcohol stress were mainly concentrated in 75 metabolic pathways, 22 genetic information processing pathways, and 8 cellular process pathways. In addition, the five pathways with a large number of differentially expressed genes were ribosomes related to genetic information (ko03010), yeast MAPK signaling pathway (ko04011), amino acid synthesis pathway (ko01230), yeast-autophagy pathway (ko04138), and oxidative phosphorylation (ko00190).

[0177] Fig.12 After KEGG annotation, the differentially expressed genes in Saccharomyces cerevisiae after 1 hour of n-dodecyl alcohol stress were found to be mainly concentrated in 77 metabolic pathways, 22 genetic information processing pathways, and 8 cellular process pathways. In addition, among the top 20 pathways with the most significant enrichment of differentially expressed genes, the five pathways with a large number of differentially expressed genes were metabolic pathway pathway (ko01100), secondary metabolism biosynthesis pathway (ko01110), amino acid synthesis pathway (ko01230), yeast MAPK signaling pathway (ko04011), and oxidative phosphorylation (ko00190).

[0178] From the KEGG enrichment analysis, the two treatment groups with different durations can affect similar pathways, such as yeast MAPK signaling pathway (ko04011) and oxidative phosphorylation (ko00190), indicating that as the stress time of n-dodecyl alcohol increases, it has a continuous impact on the MAPK signaling, amino acid synthesis, oxidative phosphorylation and other functions of Saccharomyces cerevisiae. The MAPK signaling pathway in Saccharomyces cerevisiae cells is related to spore differentiation, hyphae formation and embedded growth, high osmotic pressure glycerol formation, cell wall integrity and cell binding process. Under the stress of n-dodecyl alcohol, genes related to cell wall integrity signaling enzymes such as HKR1, WSC3, GPD2, and TUS1 were downregulated, affecting the signal transduction and maintenance of cell wall integrity. Oxidative phosphorylation occurs on the inner membrane of eukaryotic cells' mitochondria and is an important pathway for the synthesis of ATP in cells. The effect of n-dodecyl alcohol on these pathways directly affects the normal function of mitochondria in Saccharomyces cerevisiae cells.

[0179] (d) Analysis of important pathways and genes

[0180] After analysis, after 1 hour of stress on Saccharomyces cerevisiae by n-dodecyl alcohol, 36 related genes in the oxidative phosphorylation pathway were significantly upregulated compared with the control group. The top ten genes significantly upregulated in the oxidative phosphorylation pathway were determined and analyzed. The results showed that n-dodecyl alcohol can significantly upregulate the transcription level of related genes in mitochondria and the transcription level is high. These genes include succinate dehydrogenase (SDH), cytochrome C (CYC), cytochrome C oxidase (COX), mitochondrial matrix protein NDUS7, type II NADH dehydrogenase (NDH2), etc.

[0181] Succinate dehydrogenase (SDH) is a key complex enzyme in the tricarboxylic acid cycle and electron transfer respiratory chain in mitochondria. SDHx gene mutation leads to mitochondrial dysfunction, which in turn activates the pseudo-hypoxia signaling pathway. SDHA and SDHB in succinate dehydrogenase are greatly upregulated, leading to mitochondrial dysfunction. At the same time, cytochrome CCYC is also released in large quantities from mitochondria, aggravating the electron transfer disorder of the mitochondrial respiratory chain, increasing the production of ROS, and promoting cell apoptosis.

[0182] From the sequencing analysis results, it can be seen that n-dodecanol stress can significantly upregulate the transcription level of related genes in mitochondria and the transcription level is high, indicating that n-dodecanol has caused damage to cell mitochondria, which is consistent with the sharp drop in mitochondrial transmembrane potential in physiological and biochemical experiments.

[0183] From the above experimental research results on the growth, physiological and biochemical effects, and transcriptional sequencing analysis of the model organism Saccharomyces cerevisiae after stress with higher fatty alcohols, it can be seen that the higher fatty alcohols n-dodecanol and n-decanol can directly destroy the cell walls and mitochondria of fungal cells, leading to damaged cell structure, outflow of endogenous substances and mitochondrial dysfunction, causing growth inhibition, damage and death of fungal cells. This fully demonstrates that preparations containing higher fatty alcohols have antibacterial and bactericidal effects on fungi.

[0184] Example 5: Effect of aqueous emulsions containing higher fatty alcohols on the growth of Fusarium oxysporum

[0185] (1) Test reagent: The aqueous emulsion prepared by the present invention as listed in Table 2

[0186] (2) Test fungi:

[0187] Fusarium oxysporum f.sp.cubense (FOC009)

[0188] (3) Sample processing:

[0189] PDA basal culture medium: 200g potatoes, wash, peel and cut into small pieces, add 1000mL water and boil for 30min; filter with gauze, add water to the filtrate to make up to 1000mL, then add 20g glucose, and then add 20g agar to the solid to make it fully dissolved, sterilize in an autoclave at 115℃ for 20min before use.

[0190] Use a pipette to draw different components of the drug solution, add them to the unsolidified PDA culture medium after sterilization, dilute them 5000 times, 2500 times, 1000 times, 500 times and 100 times, that is, the concentrations are 0.02%, 0.04%, 0.1%, 0.2% and 1% (V / V) respectively, and then shake them thoroughly. Pour the drug-containing culture medium into 90mm culture dishes, about 15mL per dish.

[0191] Under sterile conditions, use a puncher with a diameter of 8 mm to punch out the activated fungal cake in the PDA medium. The cake is required to grow uniformly. Place the mycelium side of the cake downward and insert it into the center of the solidified PDA medium plate and culture it in a 26°C artificial climate box. After 5 days of culture, use clean water as a blank control to observe and measure the colony growth index to clarify the inhibitory effect of high-lipid membrane and different components on different fungi.

[0192] (4) Data collection and processing:

[0193] The activated fungal cakes were selected and inoculated into PDA solid culture medium and cultured in a 26°C incubator for about 5 days. The mycelium of the control group covered more than 3 / 4 of the plate.

[0194] Observe and record the mycelial growth. After 5 days, measure the colony diameter using the cross method and calculate the mycelial growth inhibition rate according to the following formula:

[0195] Mycelium growth inhibition rate = (colony growth diameter of control group - colony growth diameter of treatment group) / colony growth diameter of control group × 100%.

[0196] The effects of various agents on the culture characteristics of pathogens were compared by observing and recording the colony morphology, whether the colony edges were neat, and whether the colony surface was flat after culture at each concentration.

[0197] (5) Effects of different higher fatty alcohols on the growth of Fusarium oxysporum:

[0198] Observation under a microscope showed Fig.13 As shown in the figure, under the same concentration treatment, compared with the emulsifier control group, n-decanol and n-dodecyl alcohol have significant fungal growth inhibition ability. The colony diameter of FOC009 treated with n-decanol and n-dodecyl alcohol is small, round and milky white, and the hyphae are dense. Under other agents, the colony is round, with a large number of white aerial hyphae, irregular serrated edges, and opaque.

[0199] The growth rates of Fusarium oxysporum FOC009 after treatment with different higher fatty alcohols are shown in Table 9. It can be seen that Fusarium oxysporum FOC009 can grow under the same concentration of different higher fatty alcohols, but the growth rates under the treatment of n-decanol and n-dodecanol are lower, which are 3.28mm / d and 8.70mm / d. The colony growth rates of the emulsifier, n-hexanol, n-octane alcohol, n-tetradecanol, n-hexadecanol, n-octadecanol, n-eicosanol and n-cosadecanol treatment groups are relatively similar, all above 10.00mm / d.

[0200] The inhibition rate of higher fatty alcohols on the colony growth of FOC009 is shown in Table 10. The inhibition effect of n-decanol, n-dodecanol and n-tetradecanol treatments was better, with inhibition rates of 76.40%, 38.92% and 29.35%, respectively. The antibacterial effect of other higher fatty alcohol treatment groups was relatively general.

[0201] Table 9 Colony growth rate of FOC009 under treatment with higher fatty alcohols

[0202]

[0203] Table 10 Colony growth inhibition rate of higher fatty alcohols on FOC009

[0204]

[0205] Example 6: Effect of aqueous emulsions containing higher fatty alcohols on the growth of plant pathogenic fungi and indoor toxicity analysis

[0206] (1) Test reagent: The aqueous emulsion prepared by the present invention as listed in Table 11

[0207] Table 11 Sample No.

[0208]

[0209] The high-lipid film is a sample S aqueous emulsion containing two higher fatty alcohols, n-dodecyl alcohol and n-hexadecanol, that is, the aqueous emulsion containing higher fatty alcohols provided in Example 1, and the same below.

[0210] (2) Test fungi:

[0211] Fusarium graminearum;

[0212] Fusarium oxysporum f.sp.cubense (FOC009);

[0213] Colletotrichum gloeosporioides.

[0214] (3) Sample processing:

[0215] PDA basal culture medium: 200g potatoes, wash, peel and cut into small pieces, add 1000mL water and boil for 30min; filter with gauze, add water to the filtrate to make up to 1000mL, then add 20g glucose, and then add 20g agar to the solid to make it fully dissolved, sterilize in an autoclave at 115℃ for 20min before use.

[0216] Use a pipette to draw different components of the drug solution, add them to the unsolidified PDA culture medium after sterilization, dilute them 5000 times, 2500 times, 1000 times, 500 times and 100 times, that is, the concentrations are 0.02%, 0.04%, 0.1%, 0.2% and 1% (V / V) respectively, and then shake them thoroughly. Pour the drug-containing culture medium into 90mm culture dishes, about 15mL per dish.

[0217] Under sterile conditions, use a puncher with a diameter of 8 mm to punch out the activated fungal cake in the PDA medium. The cake is required to grow uniformly. Place the mycelium side of the cake downward and insert it into the center of the solidified PDA medium plate and culture it in a 26°C artificial climate box. After 5 days of culture, use clean water as a blank control to observe and measure the colony growth index to clarify the inhibitory effect of high-lipid membrane and different components on different fungi.

[0218] (4) Data collection and processing:

[0219] The activated fungal cakes were selected and inoculated into PDA solid culture medium and cultured in a 26°C incubator for about 5 days. The mycelium of the control group covered more than 3 / 4 of the plate.

[0220] Observe and record the mycelial growth. After 5 days, measure the colony diameter using the cross method and calculate the mycelial growth inhibition rate according to the following formula:

[0221] Mycelium growth inhibition rate = (colony growth diameter of control group - colony growth diameter of treatment group) / colony growth diameter of control group × 100%.

[0222] Calculation of toxicity regression equation and effective concentration. The toxicity of each agent to fungal mycelium was expressed as the mycelium growth inhibition rate. The mycelium growth inhibition rate was converted into the inhibition probability value (y), and the agent concentration was converted into the logarithm of the concentration (x). Virulence regression analysis was performed to calculate the EC50 and EC90 values ​​of each tested pathogenic fungus and the regression equation correlation coefficient R value.

[0223] The effects of various agents on the culture characteristics of pathogens were compared by observing and recording the colony morphology, whether the colony edges were neat, and whether the colony surface was flat after culture at each concentration.

[0224] (5) Effects of different higher fatty alcohols on the growth of Fusarium graminearum, Fusarium oxysporum, and Colletotrichum gloeosporioides:

[0225] (a) Effects of different higher fatty alcohol components on the growth of Fusarium graminearum colonies

[0226] Observation under a microscope showed Fig.14 As shown in the figure, under the treatment of 1% (V / V) dilution concentration of each component, compared with the water blank control (CK) and the emulsifier control (Emulsifier), the colony diameter of Fusarium graminearum treated with n-dodecyl alcohol was the smallest, round and light smoke pink, with a darker center, rose red, and complete and smooth edges. The colonies under the treatment of high lipid membrane (GZM) were similar to those under the treatment of n-dodecyl alcohol, but the colony growth diameter was slightly larger, the colonies were round, the center was dark rose red, surrounded by white aerial hyphae, and the edges were irregularly serrated and slightly transparent. Under the treatment of 0.2% dilution concentration of each component sample, the colonies treated with high lipid membrane and n-dodecyl alcohol had similar diameters, the center was round and orange-red, and the edges were light red; under the treatment of the other component samples, the center was round and light red or dark red, and a large number of hyphae extended around, mostly pink hyphae in the lower layer and white or yellow hyphae in the upper layer, and the edges were jagged and opaque.

[0227] The results showed that when the dilution concentration of each component sample was 1%, n-dodecyl alcohol and high lipid film had a good inhibitory effect on the mycelial growth of Fusarium graminearum. However, the emulsifier had no obvious inhibitory effect on the mycelial growth of Fusarium graminearum at different concentrations.

[0228] (b) Effects of different higher fatty alcohol components on the growth of Fusarium oxysporum colonies

[0229] Observation under a microscope showed Fig.15 As shown in the figure, under the treatment of 1% dilution concentration of each component sample, the diameter of the Fusarium oxysporum colony treated with n-dodecyl alcohol was the smallest, with a central circular milky white color, short and dense hyphae, uneven edges, and opaque. The colonies under the high lipid membrane (GZM) treatment were similar to those under the n-dodecyl alcohol treatment, but the edges of the colonies were flatter. Under the treatment of the other samples, the colonies were round, with a large number of white aerial hyphae, regular serrated edges, and slightly transparent.

[0230] The results showed that when the dilution concentration of each component sample was 1%, n-dodecyl alcohol and high lipid membrane had a good inhibitory effect on Fusarium oxysporum, and the inhibitory effect was similar. When the dilution concentration of each agent was 0.02% and 0.04%, high lipid membrane and n-dodecyl alcohol still had a certain inhibitory effect on Fusarium oxysporum.

[0231] (c) Effects of different higher fatty alcohol components on the growth of Colletotrichum gloeosporioides colonies

[0232] Observation under a microscope showed Fig.16 As shown in the figure, under the treatment of 1% dilution concentration of each component, the colony diameter of Colletotrichum gloeosporioides treated with n-dodecyl alcohol was the smallest, round and milky white, with a gray center and irregular serrated edges, and was opaque. The colonies under the high lipid membrane (GZM) treatment were similar to those under the n-dodecyl alcohol treatment. Under the treatment of the other components, the colonies were round, with a yellow-brown center, a large number of white aerial hyphae, and regular serrated edges, and were opaque.

[0233] The results showed that when the dilution concentration of each component was 1%, high lipid membrane and n-dodecyl alcohol had a better inhibitory effect on Colletotrichum gloeosporioides, and the inhibitory effects were similar.

[0234] (d) Antibacterial activity of different higher fatty alcohol components against Fusarium graminearum colonies

[0235] The growth inhibition rate of Fusarium graminearum hyphae under different treatments is shown in Table 12. Different concentrations of different higher fatty alcohol component samples have different effects on Fusarium graminearum during the culture process. Under the low concentration of 0.02%, there is no inhibitory effect on the growth of Fusarium graminearum. As the concentration increases, high lipid film, n-decanol, and n-dodecanol all have a certain inhibitory effect on the colony growth of Fusarium graminearum, and show a positive trend.

[0236] Table 12 Growth inhibition rate of Fusarium graminearum hyphae under different treatments

[0237]

[0238]

[0239] (e) Antibacterial activity of different higher fatty alcohol components against Fusarium oxysporum colonies

[0240] The growth inhibition rate of Fusarium oxysporum hyphae under different treatments is shown in Table 13. Different concentrations of different higher fatty alcohol component samples have different effects on Fusarium oxysporum during the culture process. Under the low concentration of 0.02%, the high lipid film has no inhibitory effect on the growth of Fusarium graminearum. As the concentration increases, the high lipid film, n-decanol, and n-dodecanol all have a certain inhibitory effect on the colony growth of Fusarium oxysporum, and show a positive trend.

[0241] Table 13 Growth inhibition rate of Fusarium oxysporum hyphae under different treatments

[0242]

[0243] (f) Antibacterial activity of different higher fatty alcohol components against Colletotrichum gloeosporioides colonies

[0244] The growth inhibition rate of Colletotrichum gloeosporioides hyphae under different treatments is shown in Table 14. Different concentrations of different higher fatty alcohol component samples have different effects on Colletotrichum gloeosporioides during the culture process. Under the low concentration of 0.02%, the high lipid film has no inhibitory effect on the growth of Colletotrichum gloeosporioides. As the concentration increases, the high lipid film, n-decanol, and n-dodecanol all have a certain inhibitory effect on the colony growth of Colletotrichum gloeosporioides, and show a positive trend.

[0245] Table 14 Growth inhibition rate of Colletotrichum gloeosporioides hyphae under different treatments

[0246]

[0247] (7) Toxicity analysis of different higher fatty alcohol components to the tested fungi

[0248] The indoor toxicity of different higher fatty alcohol components to three common plant pathogenic fungi was subjected to toxicity regression analysis. As shown in Table 15, except for the unstable antibacterial effect of emulsifier on the tested plant pathogenic fungi, different higher fatty alcohol components all had certain inhibitory effects on the three tested fungi. Among them, high lipid film, n-decanol and n-dodecanol had good antibacterial activity against the three tested fungi. The EC50 of high lipid film against Fusarium graminearum, Fusarium oxysporum and Colletotrichum gloeosporioides were 0.5197%, 0.3676% and 0.7115%, respectively, and the EC50 of n-decanol against Fusarium graminearum, Fusarium oxysporum and Colletotrichum gloeosporioides were 0.5657%, 0.0837% and 0.0656%, respectively. The EC50 of n-dodecanol against Fusarium graminearum, Fusarium oxysporum and Colletotrichum gloeosporioides were 0.4193%, 0.2802% and 0.7445%, respectively.

[0249] Table 15 Indoor toxicity of lipid membrane and its main component agents to three common plant pathogenic fungi

[0250]

[0251] Example 7: Effect of aqueous emulsions containing higher fatty alcohols on the growth of Peronophythora litchii and indoor toxicity analysis

[0252] (1) Test reagent: The aqueous emulsion prepared by the present invention as listed in Table 16

[0253] Table 16 Sample No.

[0254] Sample No. Sample S Sample D Sample CK (control) raw material Example 1 Implementation 2 Reagent 4 Shimizu

[0255] The high-lipid film is a sample S aqueous emulsion containing two higher fatty alcohols, n-dodecyl alcohol and n-hexadecanol.

[0256] (2) Test oomycetes:

[0257] Litchi downy mildew (Peronophythora litchii).

[0258] (3) Sample processing:

[0259] Carrot agar medium: 200g carrots, wash, peel, cut into small pieces, squeeze the juice, filter with gauze, add water to the filtrate to make up to 1000mL, then add 20g agar to fully dissolve it, sterilize in an autoclave at 115℃ for 20min before use.

[0260] Use a pipette to draw different components of the drug solution, add them to the culture medium that has not solidified after sterilization, dilute them 5000 times, 2500 times, 1000 times, 500 times and 100 times, that is, the concentrations are 0.02%, 0.04%, 0.1%, 0.2% and 1% (V / V) respectively, and then shake them thoroughly. Pour the drug-containing culture medium into 90mm culture dishes, about 15mL per dish.

[0261] Under sterile conditions, use a puncher with a diameter of 8 mm to punch out the activated oomycete cake in carrot agar medium. The cake is required to grow uniformly. Place the cake with the mycelium side facing down and insert it into the center of the solidified PDA medium plate and culture it in a 26°C artificial climate box. After 7 days of culture, use clean water as a blank control to observe and measure the colony growth index to clarify the inhibitory effect of high-lipid membrane and different components on different oomycetes.

[0262] (4) Data collection and processing:

[0263] The activated oomycete cake was selected and inoculated into carrot agar medium and cultured in a 26°C incubator for about 7 days. The mycelium of the control group covered more than 3 / 4 of the plate.

[0264] Observe and record the mycelial growth. After 7 days, measure the colony diameter using the cross method and calculate the mycelial growth inhibition rate according to the following formula:

[0265] Mycelium growth inhibition rate = (colony growth diameter of control group - colony growth diameter of treatment group) / colony growth diameter of control group × 100%.

[0266] Calculation of toxicity regression equation and effective concentration. The toxicity of each agent to oomycete hyphae was expressed as the hyphae growth inhibition rate. The hyphae growth inhibition rate was converted into the inhibition probability value (y), and the agent concentration was converted into the logarithm of the concentration (x). Virulence regression analysis was performed to calculate the EC50 and EC90 values ​​of each tested pathogenic fungus and the regression equation correlation coefficient R value.

[0267] The effects of various agents on the culture characteristics of pathogens were compared by observing and recording the colony morphology, whether the colony edges were neat, and whether the colony surface was flat after culture at each concentration.

[0268] (5) Effects of different higher fatty alcohols on the growth of litchi downy mildew:

[0269] (a) Effects of different higher fatty alcohol components on the growth of litchi Peronophytum colonies

[0270] Observation under a microscope showed Fig.17 As shown in the figure, under the treatment of 0.04%, 0.1%, 0.2% and 1% (V / V) dilution concentration of each component, compared with the water blank control (CK), the colony diameter of litchi downy mildew treated with high lipid film (sample S, GZM) and lauryalcohol ((lauryalcohol), sample D) was smaller, round, with complete and smooth edges, and the colony shape of the colony under the high lipid film treatment was similar to that under the lauryalcohol treatment. Compared with the water control, as the dilution concentration of each component sample increased, the colony diameter of the colony treated with high lipid film and lauryalcohol became smaller, indicating that the inhibitory effect on the hyphal growth of litchi downy mildew became stronger and stronger.

[0271] (b) Antibacterial activity of different higher fatty alcohol components against litchi Peronophytum colonies

[0272] The growth inhibition rate of litchi downy mildew hyphae under different treatments is shown in Table 17. Different concentrations of high lipid film and n-dodecyl alcohol treatments had different effects on litchi downy mildew during the culture process. Different concentrations of n-dodecyl alcohol and high lipid film had good inhibitory effects on the growth of litchi downy mildew hyphae, and the inhibitory effect increased with the increase of the concentration of the preparation, showing a positive trend.

[0273] Table 17 Growth inhibition rate of litchi downy mildew hyphae under different treatments

[0274]

[0275] (6) Determination of the toxicity of lipid membrane and its main component n-dodecanol to the tested oomycetes

[0276] The toxicity regression analysis of the indoor toxicity test data of high-lipid film and n-dodecyl alcohol on litchi downy mildew was performed. As shown in Table 18, both high-lipid film and n-dodecyl alcohol had a certain inhibitory effect on litchi downy mildew, and the antibacterial activity was good. The EC50 concentration of high-lipid film on litchi downy mildew was 0.0334%, and the EC50 concentration of n-dodecyl alcohol on litchi downy mildew was 0.0176%.

[0277] Table 18 Indoor toxicity of high lipid membrane and n-dodecyl alcohol pesticides to litchi downy mildew

[0278] Drug Name Oomycetes Virulence regression equation EC50(%) EC90(%) Correlation coefficient High lipid membrane Peronophytum lychee y=2.1295x+3.1423 0.0334 0.1337 0.8769 n-Dodecanol Peronophytum lychee y=1.5225x+2.6698 0.0176 0.1225 0.9644

[0279] Example 8: Preparations containing higher fatty alcohols improve the effect of preventing and controlling wheat scab

[0280] Sampling: Sample S of treatment group 1 is the aqueous emulsion prepared in Example 1; Sample C of treatment group 2 is the aqueous emulsion prepared in reagent 3 in Example 2;

[0281] Crop: Wheat Zhongken 616

[0282] Location: Caobu Town, Rudong County, Nantong City, Jiangsu Province

[0283] Method: 20 mu of treatment group 1, 20 mu of treatment group 2 and 20 mu of control group were selected in the production base for production. Treatment group 1 and treatment group 2 were produced as follows: sample S was diluted 1000 times with water and sprayed. It was sprayed once in the seedling stage, tillering stage and filling stage respectively. The sample dosage each time was 30 mL / mu. At the same time as treatment group 1 and treatment group 2, the control group was sprayed with the same amount of clean water.

[0284] Comparison of usage effects:

[0285] The wheat of treatment group 1 and treatment group 2 treated with the aqueous emulsion of the preparation containing higher fatty alcohols of the present invention have little difference in appearance, the plants grow densely, the leaves are dark green and shiny, the plants are tall, the stems are thick, and the number of tillers is more than that of the control group. The plant height, the number of leaves and the number of tillers all show obvious advantages, and the growth and appearance also have obvious advantages.

[0286] Sampling and production test results:

[0287] Yields of treatment group 1, treatment group 2 and control group were measured, and 30 ears of wheat were randomly selected from each of treatment group 1, treatment group 2 and control group to investigate and test ear length, number of grains per ear, number of empty shells, empty shell rate, susceptibility to fusarium susceptibility, 1000-grain weight, etc. The results are shown in Table 19:

[0288] Table 19 Wheat yield test and test results using the preparation containing higher fatty alcohols of the present invention

[0289] Serial number project Treatment Group 1 Treatment Group 2 Control group 1 Ear length (cm) 9.67 9.43 7.63 2 Number of grains per spike 46.10 47.4 37.83 3 Empty shell number (grains) 1.40 1.32 2.03 4 Percentage of empty shells (%) 2.95 2.78 5.10 5 Number of seeds susceptible to fusarium spores (grains) 2.53 2.62 17.43 6 Fusarium susceptibility rate (%) 5.50 5.53 46.08 7 Thousand-grain weight (g) 51.80 52.35 51.64 8 Yield per mu (kg / mu) 448.8 451.6 405.4

[0290] Fusarium graminearum is a pathogenic fungus that widely harms cereal crops. It can cause a variety of important diseases and is the main pathogen of wheat scab, affecting wheat growth. As can be seen from the above table, the control group 1 and the control group 2 using the aqueous emulsion containing higher fatty alcohols of the present invention have wheat scab susceptibility rates of 5.50% and 5.53%, which are significantly lower than the control group 46.08%. The yield measurement shows that the treatment group 1 has an acreage yield of 448.8kg, the treatment group 2 has an acreage yield of 451.6kg, and the control group has an acreage yield of 405.4kg. The yield of the treatment group has been significantly improved, with an increase of 10.71% and 11.14%, respectively. It shows that the aqueous emulsion containing higher fatty alcohols of the present invention, the treatment group 1 containing n-dodecyl alcohol, and the treatment group 2 containing n-decanol, all have a significant effect on preventing and controlling wheat scab, which is consistent with the specific antibacterial and bactericidal effect of higher fatty alcohols on Fusarium graminearum shown in Example 6.

[0291] Example 9: Effect of a preparation containing higher fatty alcohols on the prevention and control of diseases in peanut fields

[0292] Sampling: Sample S is the aqueous emulsion prepared in Example 1

[0293] Crop: Peanut Shanyou No. 35

[0294] Location: Luoba Town, Shixing County, Shaoguan City, Guangdong Province

[0295] Method: A 20-mu treatment group and a 20-mu control group were selected in the production base for production. The treatment group was produced as follows: sample S was diluted 50 times with water and mixed with seeds, and sowed after drying. Sample S was diluted 1000 times with water and sprayed. Spray once at the seedling stage, flowering stage, pod-setting stage, and maturity stage. The sample dosage each time was 15mL / mu. While the treatment group was being treated, the control group was sprayed with the same amount of clean water.

[0296] Comparison of usage effects:

[0297] The peanuts in the treatment group treated with the aqueous emulsion containing the higher fatty alcohol of the present invention had no obvious damage from diseases and insects during the entire growth process. In appearance, the plants in the treatment group grew more densely, the leaves were dark green and shiny, and the plant height was higher. Compared with the control group, the germination rate, growth condition, and the number of flower buds all showed obvious advantages.

[0298] Table 20 Peanut test results using the preparation containing higher fatty alcohols of the present invention

[0299] Serial number project Treatment Group Control group 1 Germination rate 98.6％ 91.1％ 2 Pest and disease occurrence No obvious pests and diseases 4.3% root rot 3 Rotten fruit percentage 0.94％ 11.5％

[0300] The above table shows that the peanuts treated with the preparation containing higher fatty alcohols of the present invention had no obvious pest damage and a small proportion of rotten fruits, while the control group had root rot and a large proportion of rotten fruits. The main pathogens of peanut root rot and rotten fruits are Fusarium. After treatment with the preparation containing higher fatty alcohols of the present invention, there is a significant antibacterial and bactericidal effect on Fusarium, thereby reducing the occurrence of root rot and rotten fruits in the treatment group.

[0301] Example 10: Effect of a preparation containing higher fatty alcohols on the prevention and control of diseases on Xinhui tea-branch orange trees

[0302] Sampling: Sample S is the aqueous emulsion prepared in Example 1;

[0303] Crops: Xinhui tea-branch orange;

[0304] Location: Shuangshui Town, Xinhui District, Jiangmen City, Guangdong Province;

[0305] Methods: A 20-mu treatment group and a 20-mu control group were selected at the production base for production. The treatment group was produced as follows: starting from the sprouting of tender leaves of fruit trees in spring, sample S was diluted 1000 times with water and sprayed once every 15 days, and the sample dosage each time was 100mL / mu; while the treatment group was being treated, the control group was sprayed with the same amount of clean water.

[0306] Comparison of usage effects:

[0307] The fruit trees in the treatment group treated with the aqueous emulsion containing the higher fatty alcohol of the present invention are strong and vigorous, with dark green and shiny leaves, no citrus anthracnose is found, and the fruit drop is small.

[0308] Table 21 Test results of citrus fruits using the preparation containing higher fatty alcohols of the present invention

[0309]

[0310] The above table shows that the fruit trees treated with the preparation containing higher fatty alcohols of the present invention had no obvious pest damage and less fruit drop, while the control group had anthracnose and a high proportion of fruit drop. Citrus anthracnose is caused by infection by Aspergillus niger, which can harm the leaves, treetops, flowers and fruits of fruit trees. After treatment with the preparation containing higher fatty alcohols of the present invention, it has a significant antibacterial and bactericidal effect on Aspergillus niger, thereby reducing the occurrence of anthracnose and physiological fruit drop in the treatment group.

[0311] Example 11: Effect of formulations containing higher fatty alcohols on preventing and controlling diseases in sugarcane

[0312] Sampling: Sample S of treatment group 1 is the aqueous emulsion prepared in Example 1; Sample C of treatment group 2 is the aqueous emulsion prepared in reagent 3 in Example 2;

[0313] Crops: Sugarcane Guitang No. 44;

[0314] Location: Luobai Community, Jiangzhou District, Chongzuo City, Guangxi;

[0315] Method: 50 mu of treatment group 1, 50 mu of treatment group 2 and 50 mu of control group were selected in the production base for production. Treatment group 1 and treatment group 2 were produced as follows: sample S was diluted 1000 times with water and sprayed once every 7 days, and the sample dosage each time was 100 mL / mu; while treatment group 1 and treatment group 2 were being treated, the control group was sprayed with the same amount of clean water.

[0316] Comparison of usage effects:

[0317] The sugarcanes treated with the aqueous emulsion containing higher fatty alcohols of the present invention had similar appearances between treatment group 1 and treatment group 2. The leaves of the plants were dark green and shiny, and the growth was vigorous. However, the leaves of the plants in the control group were somewhat yellowish, and the growth was weak.

[0318] 30 square meters were randomly selected as survey points in treatment group 1, treatment group 2 and the control group to investigate the number of smut and count the changes in the number of smut, as shown in Table 22.

[0319] Table 22 Effect of the preparation containing higher fatty alcohols of the present invention on the prevention and treatment of sugarcane smut

[0320]

[0321] The above table shows that the leaves of the sugarcanes in treatment group 1 and treatment group 2 treated with the preparation containing higher fatty alcohols of the present invention are dark green and shiny, and grow vigorously, while the leaves of the control group are somewhat yellowish and grow weakly. After treatment, the increase in the number of sugarcane smut in treatment group 1 and treatment group 2 is small, the growth rate is low, and the smut is obviously suppressed, while the smut in the control group is increasing, and the growth rate is also getting faster and faster; the main pathogen of sugarcane smut is sugarcane whip smut, which is the most important fungal disease in sugarcane planting and one of the most common and most serious diseases in sugarcane cultivation. The use of the preparation containing higher fatty alcohols of the present invention, whether it is treatment group 1 containing n-dodecanol or treatment group 2 containing n-decanol, has obvious antibacterial and bactericidal effects on fungi after treatment, thereby reducing the occurrence of sugarcane smut in the treatment groups.

[0322] Example 12: Effect of a preparation containing higher fatty alcohols on preventing and controlling diseases in cucumber fields

[0323] Sampling: Sample S is the aqueous emulsion prepared in Example 1;

[0324] Crop: Cucumber 8808;

[0325] Location: Shuqiao Town, Daming County, Handan City, Hebei Province;

[0326] Method: A 5-mu treatment group and a 5-mu control group were selected in the production base for production. The treatment group was produced as follows: Sample S was diluted 100 times with water to soak the roots of cucumber seedlings before planting, and sample S was diluted 800 times with water and sprayed. Spray once at the seedling stage, flowering stage, and harvest stage, and the sample dosage each time was 25mL / mu. While the treatment group was being treated, the control group was sprayed with the same amount of clean water.

[0327] Comparison of usage effects:

[0328] The cucumbers in the treatment group treated with the aqueous emulsion containing the higher fatty alcohol of the present invention had no obvious damage from diseases and insect pests during the entire growth process. In appearance, the plants in the treatment group grew better, had lush leaves, and showed significant advantages in the number of flowers and fruits, and maintained better tree vigor.

[0329] Table 23 Cucumber test results using the preparation containing higher fatty alcohols of the present invention

[0330] Serial number project Treatment Group Control group 1 Yield per mu (kg / mu) 1663 845 2 Cucumber powdery mildew No findings Powdery mildew was found during the flowering period, with a disease rate of 23% 3 Cucumber vine blight No findings Vine blight was found in the seedling stage, with a disease rate of 18%

[0331] The above table shows that the cucumbers treated with the preparation containing higher fatty alcohols of the present invention had good growth throughout the growth process, no obvious diseases and insect pests were found, and the yield was significantly increased by 96.8%. In the control group, powdery mildew and vine blight were found throughout the growth process. The cucumber powdery mildew and vine blight harmed the leaves and vines, affecting the normal growth of cucumbers and seriously affecting the yield. The cucumber vine blight was caused by Ascochyta citrinum and Mycosphaeria melonea, and the cucumber powdery mildew was caused by the infection of P. leucoderma and P. citrinum melonea. Ascochyta citrinum, Mycosphaeria melonea, P. leucoderma and P. leucoderma melonea all belong to the subphylum Ascomycota. Obviously, the preparation containing higher fatty alcohols of the present invention has obvious antibacterial and bactericidal effects on fungi. The use of the treatment group after treatment reduced the occurrence of cucumber powdery mildew and vine blight, and significantly increased the cucumber yield.

[0332] Example 13: Effect of a preparation containing higher fatty alcohols on the prevention and control of diseases in eggplant fields

[0333] Sampling: Sample S is the aqueous emulsion prepared in Example 1;

[0334] Crop: Long Eggplant Black Dragon Supreme No. 2;

[0335] Location: Shuqiao Town, Daming County, Handan City, Hebei Province;

[0336] Method: A 2-mu treatment group and a 2-mu control group were selected in the production base for production. The treatment group was produced as follows: sample S was diluted 100 times with water to soak the roots of eggplant seedlings before planting, and sample S was diluted with water and sprayed. Spray once in the seedling stage, flowering stage, fruiting stage and harvesting stage. The dilution multiples in the seedling stage, flowering stage, fruiting stage and harvesting stage were 800 times, 600 times, 500 times and 500 times, and the sample dosage each time was 60 mL / mu. While the treatment group was being treated, the control group was sprayed with the same amount of clean water.

[0337] Comparison of usage effects:

[0338] The eggplants in the treatment group treated with the aqueous emulsion of the preparation containing higher fatty alcohols of the present invention have no obvious damage from diseases and pests during the entire growth process. In terms of appearance, the plants in the treatment group have stronger growth conditions, wider leaves, more flowers and fruits per plant, better appearance and more uniform coloring of the fruits, all of which show significant advantages.

[0339] Table 24 Eggplant test results using the preparation containing higher fatty alcohols of the present invention

[0340] Serial number project Treatment Group Control group 1 Yield per mu (kg / mu) 1617 1284 2 Eggplant Botrytis cinerea No findings Gray mold was found in the seedling and fruiting stages, with a disease rate of 14.8% 3 Eggplant blight No findings Blight was found in the fruiting period, with a disease rate of 4.5%

[0341] The above table shows that the eggplants treated with the preparation containing higher fatty alcohols of the present invention have no obvious diseases and insect pests, the plants are strong, the fruits are abundant, and the yield is increased by 25.9% compared with the control group. In the whole growth process, the control group was found to have eggplant gray mold and woolly blight. The pathogen of eggplant gray mold is Botrytis cinerea, which belongs to the subphylum Ascomycota. The pathogen of eggplant woolly blight is Phytophthora nicotianae, which belongs to the genus Phytophthora of the phylum Oomycetes of the algae kingdom. The eggplant is attacked by these two diseases, resulting in a decrease in yield. By comparison, it can be seen that the preparation containing higher fatty alcohols of the present invention has obvious antibacterial and bactericidal effects on fungi after treatment, thereby reducing the occurrence of eggplant gray mold and woolly blight in the treatment group and significantly improving the eggplant yield.

[0342] Example 14: Effect of a preparation containing higher fatty alcohols on preventing and controlling diseases in cowpea fields

[0343] Sampling: Sample S of treatment group 1 is the aqueous emulsion prepared in Example 1; Sample C of treatment group 2 is the aqueous emulsion prepared in reagent 3 in Example 2;

[0344] Crops: Cowpea, large meat cowpea;

[0345] Location: Sanxiang Town, Zhongshan City, Guangdong Province;

[0346] Methods: 2 mu of treatment group 1, 2 mu of treatment group 2 and 2 mu of control group were selected in the production base for production. Treatment group 1 and treatment group 2 were produced as follows: sample S was diluted 100 times with water to soak the roots of cowpea seedlings before planting, and sample S was diluted with water and sprayed. The seedling stage, flowering stage, fruiting stage and harvesting stage were sprayed once each. The dilution multiples in the seedling stage, flowering stage, fruiting stage and harvesting stage were 800 times, 600 times, 500 times and 500 times, and the sample dosage each time was 60 mL / mu. While the treatment group was being treated, the control group was sprayed with the same amount of clean water.

[0347] Comparison of usage effects:

[0348] The cowpeas in treatment groups 1 and 2 treated with the aqueous emulsion containing higher fatty alcohols of the present invention had good plant growth, lush leaves, and significant advantages in the number of flowers and pods during the entire growth process, and the number of flowers and pods was even greater in the late harvest period.

[0349] Table 25 Test results of cowpea preparation containing higher fatty alcohols of the present invention

[0350]

[0351] The above table shows that the cowpea treated with the preparation containing higher fatty alcohols of the present invention has good plant growth, a large number of flowering and pods, and a high yield. The per-acre yields of treatment group 1 and treatment group 2 are 3297kg and 3184kg, respectively, which are 17.8% and 13.8% higher than the per-acre yield of the control group of 2798kg; at the same time, the incidence of diseases in the treatment group is low, while the control group is more affected by cowpea damping-off and cowpea rust during the entire growth process. The pathogen of cowpea rust is cowpea unicellular rust fungus, which belongs to the subphylum Basidiomycetes. The pathogen of cowpea damping-off is mainly Rhizoctonia solani, which belongs to the subphylum Ascomycota. Cowpea is affected by these two diseases, resulting in reduced yield. By comparison, it can be seen that the preparation containing higher fatty alcohols of the present invention has obvious antibacterial and bactericidal effects on these two fungi. The use of the post-treatment group reduces the occurrence of cowpea damping-off and cowpea rust, and significantly improves the yield of cowpea.

[0352] Example 15: Effect of a preparation containing higher fatty alcohols on the prevention and control of diseases in pepper fields

[0353] Sampling: Sample S is the aqueous emulsion prepared in Example 1;

[0354] Crops: Processed pepper Fuguihong;

[0355] Location: Karamay Agricultural Development Zone, Xinjiang;

[0356] Method: A 40-mu treatment group and a 40-mu control group were selected at the production base for production. The treatment group was produced as follows: Sample S was diluted 800 times with water and sprayed, spraying once every 15 days starting from the seedling stage, and the sample dosage each time was 60mL / mu; while the treatment group was being treated, the control group was sprayed with the same amount of clean water.

[0357] Comparison of usage effects:

[0358] The peppers in the treatment group treated with the aqueous emulsion containing the higher fatty alcohol of the present invention were free from disease damage during the entire growth process. The plants were strong in appearance, with thick and vigorous branches and trunks. The new and old leaves had a good degree of expansion, the leaves were light green, and the peppers were full of the plants.

[0359] Table 26 Pepper test results using the preparation containing higher fatty alcohols of the present invention

[0360] Serial number project Treatment Group Control group 1 Yield per mu (kg / mu) 850.1 690.0 2 Pepper blight No findings Pepper blight was found in the seedling and mature stages, with a disease rate of 11.3%

[0361] The above table shows that the peppers treated with the preparation containing higher fatty alcohols of the present invention have strong plants, more fruits, and high yields, which is 23.2% higher than the control group, and there are no obvious diseases and insect pests, while the control group is attacked by pepper blight during the entire growth process. The pathogen of pepper blight is pepper blight, which belongs to the genus Phytophthora of the subphylum Mastophora. When attacked by pepper blight, the growth and yield of peppers are seriously affected. By comparison, it can be seen that the preparation containing higher fatty alcohols of the present invention has a significant antibacterial and bactericidal effect on this fungus. The post-treatment group reduces the occurrence of pepper blight and significantly increases the yield of pepper.

[0362] Example 16: Effect of a preparation containing higher fatty alcohols on preventing and controlling diseases in apple fields

[0363] Sampling: Sample S is the aqueous emulsion prepared in Example 1;

[0364] Crops: Apple Red Apple;

[0365] Location: Karamay Agricultural Development Zone, Xinjiang;

[0366] Method: A 20-mu treatment group and a 20-mu control group were selected in the production base for production. The treatment group was produced as follows: sample S was diluted with water and sprayed, 800 times in the young fruit stage and 600 times in the swelling fruit stage, once every 15 days, and the sample dosage each time was 100mL / mu; while the treatment group was being treated, the control group was sprayed with the same amount of clean water.

[0367] Comparison of usage effects:

[0368] The apples in the treatment group treated with the aqueous emulsion containing the higher fatty alcohol of the present invention are free from disease damage during the entire growth process, and the number of fruits on the plants is more, the apples are healthier and more lustrous in appearance, the quality is better, the number of rotten fruits and deformed fruits is less, and the fruits of the plants in the treatment group change color earlier.

[0369] Table 27 Apple test results using the preparation containing higher fatty alcohols of the present invention

[0370] Serial number project Treatment Group Control group 1 Yield per mu (kg / mu) 1985 1677 2 Apple tree rot No obvious disease was found Apple tree rot was found, with a disease rate of 9.2%

[0371] The above table shows that the apples treated with the preparation containing higher fatty alcohols of the present invention have strong plants, more fruits, fewer rotten fruits and deformed fruits, and high yields, which is 18.4% higher than the control group, and there are no obvious diseases and insect pests, while the control group has a certain degree of apple tree rot during the entire growth process. The pathogen of apple tree rot is black rot fungus of apple, which belongs to the subdivision of Ascomycota. When attacked by apple tree rot, the growth of apple trunks and fruits is seriously affected. By comparison, it can be seen that the preparation containing higher fatty alcohols of the present invention has a significant antibacterial and bactericidal effect on this fungus. The post-treatment group reduces the damage of apple tree rot and significantly increases the yield of apples.

[0372] Example 17: Effect of a preparation containing higher fatty alcohols on preventing and controlling diseases on litchi trees

[0373] Sampling: Sample S of treatment group 1 is the aqueous emulsion prepared in Example 1; Sample C of treatment group 2 is the aqueous emulsion prepared in reagent 3 in Example 2;

[0374] Crop: Lychee Guiwei;

[0375] Location: Dajing Town, Gaozhou City, Guangdong Province;

[0376] Method: 10 mu of treatment group 1, 10 mu of treatment group 2 and 10 mu of control group were selected in the production base for production. Treatment group 1 and treatment group 2 were produced as follows: starting from the sprouting of tender leaves of fruit trees in spring, the sample was diluted 1000 times with water and sprayed, and sprayed once every 15 days. The sample dosage each time was 100 mL / mu; while the treatment group was being treated, the control group was sprayed with the same amount of clean water.

[0377] Comparison of usage effects:

[0378] The fruit trees in the treatment group treated with the aqueous emulsion containing the higher fatty alcohol of the present invention were strong, with dark green and shiny leaves, no obvious pests and diseases, and few cracked and rotten fruits. However, the fruit trees in the control group were found to have litchi anthracnose and litchi downy mildew, and some fruit trees were weak, with many fallen leaves and fruits and many cracked and rotten fruits.

[0379] Table 28 Litchi test results using the preparation containing higher fatty alcohols of the present invention

[0380]

[0381] The above table shows that the fruit trees treated with the preparation containing higher fatty alcohols of the present invention grow vigorously and have less cracked and rotten fruits. The per-acre yield of litchi in treatment group 1 and treatment group 2 are 314kg and 326kg, respectively, which are 28.2% and 33.1% higher than 245kg of the control group. Both treatment groups have no obvious pest damage, while the control group has anthracnose and downy mildew. The litchi anthracnose is caused by infection of Colletotrichum gloeosporioides or Colletotrichum gloeosporioides, which are the direct causes of the disease, while the pathogen of litchi downy mildew is Peronophythora litchii, which is an oomycete that can affect the young leaves, young branches, flower spikes and fruits of litchi, causing significant yield losses. The preparation containing the higher fatty alcohol (n-decanol or n-dodecyl alcohol) of the present invention is used for treatment. No matter it is the fungus that causes litchi anthracnose or the oomycete that causes litchi downy mildew, it has obvious antibacterial and bactericidal effects, thereby reducing the occurrence of litchi anthracnose and litchi downy mildew in the treatment group and reducing possible yield losses.

[0382] From the experimental research results of the growth, physiological and biochemical effects, and transcriptional sequencing analysis of the model organism Saccharomyces cerevisiae after stress with higher fatty alcohols in Example 4, it can be seen that higher fatty alcohols destroy the integrity of the cell wall of Saccharomyces cerevisiae cells, resulting in damaged cell structure and outflow of endogenous substances. At the same time, the mitochondrial function of Saccharomyces cerevisiae cells is destroyed and the transcription of key genes in the oxidative phosphorylation pathway is upregulated, thereby causing cell damage and death, indicating that higher fatty alcohols have inhibitory and killing effects on eukaryotic cell organisms.

[0383] It can be seen from Examples 5 and 6 that in laboratory experiments, it was determined that the preparation containing higher fatty alcohols of the present invention had a certain inhibitory and killing effect on the growth of plant pathogenic fungi such as Fusarium graminearum, Fusarium oxysporum and Colletotrichum gloeosporioides. These results are consistent with the results of the effect of higher fatty alcohols on the growth of the model organism Saccharomyces cerevisiae in Example 4, indicating that higher fatty alcohols can directly destroy the cell walls and mitochondria of fungal cells, resulting in damage to the cell structure, outflow of endogenous substances and dysfunction of mitochondria, causing growth inhibition, damage and death of fungal cells, indicating that higher fatty alcohols have antibacterial and bactericidal effects on plant pathogenic fungi.

[0384] As can be seen from Example 7, it was determined in laboratory tests that the preparation containing higher fatty alcohols of the present invention has a certain inhibitory and killing effect on the growth of the plant pathogenic oomycete Peronophytum litchiensis.

[0385] It can be seen from Examples 8 to 17 that the use of the emulsifier containing higher fatty alcohols of the present invention on various crops such as wheat, peanuts, citrus, sugarcane, cucumber, eggplant, cowpea, pepper, apple, litchi, etc. can effectively prevent and control various plant diseases caused by plant pathogenic fungi and oomycetes, and increase yield. This shows that the use of the emulsifier containing higher fatty alcohols of the present invention can inhibit and kill fungi and oomycetes, and effectively prevent and control related diseases caused by plant pathogenic fungi and oomycetes.

[0386] Based on Examples 4 to 17, the application of the preparation containing higher fatty alcohols of the present invention can directly destroy the cell wall and mitochondria of fungal cells, resulting in damage to the cell structure, outflow of endogenous substances and dysfunction of mitochondria, causing growth inhibition, damage and death of fungal cells. At the same time, the preparation containing higher fatty alcohols can also inhibit the growth of oomycetes, indicating that the preparation containing higher fatty alcohols has antibacterial and bactericidal effects on fungi and oomycetes. Using the preparation containing higher fatty alcohols of the present invention on plants can inhibit and kill plant pathogenic fungi and oomycetes, thereby effectively preventing and controlling diseases caused by plant pathogenic fungi and oomycetes.

[0387] The above-mentioned embodiments are only preferred embodiments of the present invention and cannot be used to limit the scope of protection of the present invention. Any non-substantial changes and substitutions made by technicians in this field on the basis of the present invention shall fall within the scope of protection required by the present invention.

Claims

1. The use of higher fatty alcohols in the preparation of fungi-inhibiting and bactericidal preparations, characterized in that: The higher fatty alcohol includes n-decanol, n-dodecyl alcohol or a mixture of the two.

2. The use according to claim 1, characterized in that The preparation achieves the application of inhibiting and killing fungi by destroying the integrity of cell walls, resulting in damage to cell structure and outflow of endogenous substances, causing growth inhibition, damage and death of fungal cells.

3. The use according to claim 1, characterized in that The preparation achieves its application in inhibiting and killing fungi by causing mitochondrial function destruction and transcriptional upregulation of key genes in the oxidative phosphorylation pathway, thereby causing growth inhibition, damage and death of fungal cells.

4. The use according to any one of claims 1 to 3, characterized in that The preparation includes one of aqueous emulsion, emulsifiable concentrate, suspension, wettable powder, dust, granule, tablet, aqueous solution, sprayable solution, concentrated emulsion, aerosol and seed coating agent.

5. The use of higher fatty alcohols in the preparation of preparations for the prevention and control of diseases caused by plant pathogenic fungi, characterized in that: The higher fatty alcohol includes n-decanol, n-dodecyl alcohol or a mixture of the two.

6. The use according to claim 5, characterized in that The preparation achieves prevention and control of diseases caused by plant pathogenic fungi by destroying the integrity of the cell wall of the plant pathogenic fungi, resulting in damage to the cell structure, outflow of endogenous substances, and causing growth inhibition, damage and death of fungal cells.

7. The use according to claim 5, characterized in that The preparation achieves prevention and control of diseases caused by plant pathogenic fungi by causing mitochondrial function destruction and transcriptional upregulation of key genes in the oxidative phosphorylation pathway, thereby causing growth inhibition, damage and death of fungal cells.

8. The use according to any one of claims 5 to 7, characterized in that: The preparation includes one of aqueous emulsion, emulsifiable concentrate, suspension, wettable powder, dust, granule, tablet, aqueous solution, sprayable solution, concentrated emulsion, aerosol and seed coating agent.

9. Use of higher fatty alcohols in the preparation of bacteriostatic and bactericidal preparations for oomycetes, characterized in that: The higher fatty alcohol includes n-decanol, n-dodecanol or a mixture of the two.

10. Use of higher fatty alcohols in the preparation of a preparation for the prevention and control of diseases caused by plant pathogenic oomycetes, characterized in that: The higher fatty alcohol includes n-decanol, n-dodecyl alcohol or a mixture of the two.

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