Use of higher fatty alcohol for antimicrobial purpose

Abstract

A use of a higher fatty alcohol in preparation of a fungistatic and fungicidal formulation, a use of a higher fatty alcohol in preparation of a formulation for controlling diseases caused by plant pathogenic fungi, a use of a higher fatty alcohol in preparation of an oomycetostatic and oomyceticidal formulation, and a use of a higher fatty alcohol in preparation of a formulation for controlling diseases caused by plant pathogenic oomycetes, wherein the higher fatty alcohol includes one or a mixture of n-decanol and n-dodecanol.

Description

Application of higher aliphatic alcohol in bacteriostasis and fungicidal aspects TECHNICAL FIELD

[0001] The application belongs to the technical field of crop disease and pest control, and particularly relates to application of higher aliphatic alcohol in bacteriostasis and fungicidal aspects. BACKGROUND

[0002] It is estimated that the global crop yield loss caused by diseases and pests is equivalent to one-third of the potential harvest amount each year, and the diseases caused by plant pathogenic fungi and oomycetes account for more than 70% of the total plant diseases. At present, the plant diseases are mainly controlled by chemical agents, which have played a role in prevention and control in a short time, but the resistance risk, pesticide residues and environmental pollution and other problems caused by long-term single application of chemical agents have become increasingly prominent. Reducing the risk of pesticide resistance and cross-resistance, creating efficient, environmentally safe and green new pesticides has become a new trend of global pesticide research and development.

[0003] Meanwhile, 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 leading to the failure of disease control. The development of agents that can simultaneously inhibit both types of pathogens has very important value in agricultural production.

[0004] Studies have shown that the growth of fungi 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 leading to cell death.

[0005] Higher aliphatic alcohol is composed of a fatty chain with multiple carbon atoms and a hydroxyl group at one end, and has a specific carbon chain length in chemical structure, often has certain solubility and crystallinity, and has characteristics such as low toxicity, low irritation and non-flammability, which makes them a kind of green and environmentally friendly chemicals.

[0006] SUMMARY

[0007] In order to effectively prevent and control the invasion of plant pathogenic fungi and oomycetes on crops, the application provides application of higher aliphatic alcohol in bacteriostasis and fungicidal aspects.

[0008] To solve the above problems, the technical scheme adopted by the application is as follows:

[0009] The application provides application of higher aliphatic alcohol in preparation of a bacteriostatic and fungicidal agent for fungi, wherein the higher aliphatic alcohol includes one of n-decanol and n-dodecanol or a mixture of the two.

[0010] Further, the preparation achieves the application of inhibiting and killing fungi and oomycetes by destroying the cell wall integrity, causing the cell structure to be damaged, the endogenous substances to outflow, and causing the growth inhibition, damage and death of the fungal cells.

[0011] Further, the preparation achieves the application of inhibiting and killing fungi by destroying the mitochondrial function and the transcription up-regulation of the key genes of the oxidative phosphorylation pathway, causing the growth inhibition, damage and death of the fungal cells.

[0012] In the present application, the preparation includes but is not limited to the water emulsion, the emulsifiable concentrate, the suspension, the wettable powder, the powder, the granule, the water agent, the sprayable solution, the concentrated emulsion, the aerosol, the seed coating agent and the like, as long as all the dosage forms using the inventive concept of the present application are applicable.

[0013] In another aspect, the present application provides the application of the higher fatty alcohol in the preparation of the preparation for preventing and controlling the disease caused by the plant pathogenic fungi, wherein the higher fatty alcohol includes one of n-decanol and n-dodecanol or the mixture of the two.

[0014] Further, the preparation achieves the application of preventing and controlling the disease caused by the plant pathogenic fungi by destroying the cell wall integrity of the plant pathogenic fungi, causing the cell structure to be damaged, the endogenous substances to outflow, and causing the growth inhibition, damage and death of the fungal cells.

[0015] Further, the preparation achieves the application of preventing and controlling the disease caused by the plant pathogenic fungi by destroying the mitochondrial function and the transcription up-regulation of the key genes of the oxidative phosphorylation pathway, causing the growth inhibition, damage and death of the fungal cells.

[0016] In the present application, the preparation includes but is not limited to the water emulsion, the emulsifiable concentrate, the suspension, the wettable powder, the powder, the granule, the water agent, the sprayable solution, the concentrated emulsion, the aerosol, the seed coating agent and the like, as long as all the dosage forms using the inventive concept of the present application are applicable.

[0017] In another aspect, the present application provides the application of the higher fatty alcohol in the preparation of the preparation for inhibiting and killing the oomycetes, wherein the higher fatty alcohol includes one of n-decanol and n-dodecanol or the mixture of the two.

[0018] In another aspect, the present application provides the application of the higher fatty alcohol in the preparation of the preparation for preventing and controlling the disease caused by the plant pathogenic oomycetes, wherein the higher fatty alcohol includes one of n-decanol and n-dodecanol or the mixture of the two.

[0019] In the present application, the preparation includes but is not limited to water emulsion, emulsifiable concentrate, suspension, wettable powder, powder, granule, water agent, sprayable solution, concentrated emulsion, aerosol, seed coating agent, etc. as long as all dosage forms using the inventive concept of the present application are applicable.

[0020] Further, the preparation is a water emulsion, which includes the higher fatty alcohol, an emulsifier, a thickening agent, and water.

[0021] Compared with the prior art, the effect of the present application is that:

[0022] The present application uses advanced physiological and biochemical detection and transcriptome sequencing analysis technology to screen and determine the preparation containing higher fatty alcohol, which can directly destroy the cell wall and mitochondria of fungal cells, cause cell structure damage, endogenous substance outflow and mitochondrial dysfunction, cause growth inhibition, damage and death of fungal cells, and achieve the purpose of inhibiting and killing bacteria, so as to realize the prevention and control of diseases caused by plant pathogenic fungi. At the same time, higher fatty alcohol can also inhibit the growth of oomycetes and has the effect of inhibiting and killing bacteria, so as to realize the prevention and control of diseases caused by plant pathogenic oomycetes. The present application applies higher fatty alcohol to the inhibition and sterilization of crops, and provides a green, environmentally friendly and efficient prevention and control method for diseases caused by plant pathogenic fungi and oomycetes, which is a new choice for crop disease prevention and control. BRIEF DESCRIPTION OF DRAWINGS

[0023] Fig. 1 is a diagram of the mycelial growth of Saccharomyces cerevisiae treated with various higher fatty alcohols and emulsifiers;

[0024] Fig. 2 is a SEM diagram of Saccharomyces cerevisiae cells treated with an emulsifier;

[0025] Fig. 3 is a SEM diagram of Saccharomyces cerevisiae cells treated with n-decanol;

[0026] Fig. 4 is a SEM diagram of Saccharomyces cerevisiae cells treated with n-dodecanol;

[0027] Fig. 5 is a single-parameter histogram of cell integrity flow cytometry detection;

[0028] Fig. 6 is a single-parameter histogram of cell ROS flow cytometry detection;

[0029] Fig. 7 is a volcano plot of differentially expressed genes of Saccharomyces cerevisiae cells treated with n-dodecanol and emulsifier for 5 min;

[0030] Fig. 8 is a volcano plot of differentially expressed genes of Saccharomyces cerevisiae cells treated with n-dodecanol and emulsifier for 1 h;

[0031] Fig. 9 is a GO enrichment analysis diagram of differentially expressed genes of Saccharomyces cerevisiae cells treated with n-dodecanol and emulsifier for 5 min;

[0032] Figure 10 is a GO enrichment analysis chart of differentially expressed genes of yeast cells after dodecanol and emulsifier treatment for 1 h;

[0033] Figure 11 is a KEGG enrichment analysis chart of differentially expressed genes of yeast cells after dodecanol and emulsifier treatment for 5 min;

[0034] Figure 12 is a KEGG enrichment analysis chart of differentially expressed genes of yeast cells after dodecanol and emulsifier treatment for 1 h;

[0035] Figure 13 is a diagram of the mycelial growth of Fusarium oxysporum FOC009 after 5 d of treatment with higher aliphatic alcohols;

[0036] Figure 14 is a diagram of the mycelial growth of Fusarium graminearum after 5 d of treatment with different higher aliphatic alcohols;

[0037] Figure 15 is a diagram of the mycelial growth of Fusarium oxysporum after 5 d of treatment with different higher aliphatic alcohols;

[0038] Figure 16 is a diagram of the mycelial growth of Colletotrichum gloeosporioides after 5 d of treatment with different higher aliphatic alcohols;

[0039] Figure 17 is a diagram of the mycelial growth of Peronophythora litchii after 7 d of treatment with different higher aliphatic alcohols. DETAILED DESCRIPTION

[0040] In order to make the skilled in the art more clearly understand the technical solutions described in the present application, the following examples are listed for illustration. It should be pointed out that the following examples do not constitute a limitation on the scope of protection required by the present application.

[0041] The raw materials, reagents or devices used in the following examples, unless otherwise specified, can be obtained from conventional commercial channels, or can be obtained by existing known methods.

[0042] Example 1

[0043] The present example provides a higher aliphatic alcohol-containing water emulsion, which consists of the following components by weight percentage: dodecanol 40%, n-hexadecanol 5%, emulsifier 4%, thickening agent 3%, and water in the remainder.

[0044] In the present example, the emulsifier is castor oil polyoxyethylene ether, and the thickening agent is carboxymethyl cellulose.

[0045] The present example also provides a preparation method of the above-mentioned water emulsion, comprising the following steps:

[0046] Put n-dodecanol, n-hexadecanol into a container and heat to 55°C to melt, then add emulsifier in the container, process by high shear homogenizer, rotate at 4500 rpm for 10 min; then add 55°C water in the container, process by high shear homogenizer, rotate at 10000 rpm for 20 min, then cool to 40°C; then add thickening agent in the container, process by high shear homogenizer, rotate at 10000 rpm for 30 min, then obtain.

[0047] Example 2

[0048] The example provides a water emulsion containing higher fatty alcohol, which consists of the following components by weight percentage: fatty alcohol 40%, emulsifier 4%, thickening agent 3%, and water in remainder.

[0049] In the example, the emulsifier is castor oil polyoxyethylene ether, and the thickening agent is carboxymethyl cellulose.

[0050] In the example, the reagents and fatty alcohols correspond to Table 1:

[0051] Table 1: Selection of fatty alcohols in each reagent

[0052] The example also provides a preparation method of the above water emulsion, which comprises the following steps:

[0053] Put fatty alcohol into a container and heat to 75°C to melt, then add emulsifier in the container, process by high shear homogenizer, rotate at 4500 rpm for 10 min; then add 75°C water in the container, process by high shear homogenizer, rotate at 10000 rpm for 20 min, then cool to 40°C; then add thickening agent in the container, process by high shear homogenizer, rotate at 10000 rpm for 30 min, then obtain.

[0054] Example 3

[0055] The example provides a water emulsion, which consists of the following components by weight percentage: emulsifier 4%, thickening agent 3%, and water in remainder.

[0056] In the example, the emulsifier is castor oil polyoxyethylene ether, and the thickening agent is carboxymethyl cellulose.

[0057] Put emulsifier into a container and heat to 55°C, process by high shear homogenizer, rotate at 4500 rpm for 10 min, then add 55°C water in the container, process by high shear homogenizer, rotate at 10000 rpm for 20 min, then cool to 40°C, then add thickening agent in the container, process by high shear homogenizer, rotate at 10000 rpm for 30 min, then obtain.

[0058] Example 4

[0059] Effect of water emulsion containing higher aliphatic alcohols on growth of Saccharomyces cerevisiae and mechanism research

[0060] Saccharomyces cerevisiae is a very important yeast in human life, and the cell morphology is mainly spherical or oval, which mainly reproduces by budding. It can survive under aerobic and anaerobic conditions, and has strong adaptability. Saccharomyces cerevisiae, as a single-cell eukaryotic microorganism, has high similarity in cell and organelle structure with animal and plant cells. The genes and biochemical pathways found in Saccharomyces cerevisiae are highly conserved in higher organisms, and it is often used as a model organism for studying eukaryotes, and has significant genetic applicability advantages.

[0061] Using Saccharomyces cerevisiae as a model organism to study the bacteriostatic and bactericidal mechanism of higher aliphatic alcohols on fungi and oomycetes has great universality.

[0062] (1) Effect of water emulsion containing higher aliphatic alcohols on growth of Saccharomyces cerevisiae

[0063] Test agent: water emulsion prepared according to Table 2.

[0064] Table 2 Sample No.

[0065] (2) Test fungi:

[0066] Saccharomyces cerevisiae W303-1A.

[0067] (3) Test method

[0068] Preparation of YPD base medium: yeast extract 10g, tryptone 20g, glucose 20g, solid agar powder 20g, add water to 1000mL. 115℃, sterilize for 20min.

[0069] Pick the W303-1A strain preserved at-80℃ and streak on YPD solid plate, and incubate in 30℃ incubator for 3-5d until single colonies appear.

[0070] Pick single colonies and inoculate in 5mL YPD liquid medium, incubate at 30℃, 220rpm for 12-16h.

[0071] Take 1mL of bacterial solution and add 50mL of YPD liquid medium, incubate at 30℃, 220rpm for 12-16h.

[0072] The sample was taken 1 mL, and the absorbance was measured at OD600. The concentration of each sample was adjusted using deionized water to dilute, and the value OD600 = 1.0.

[0073] 100 μL of the above diluted bacteria solution with OD600 = 1.0 was added to the first tube of eight PCR tubes, and then 90 μL of sterilized deionized water was added to the five tubes. 10 μL of the bacteria solution was taken from the first tube and added to the second tube, which was mixed by blowing, and then 10 μL of the solution was taken from the second tube and added to the third tube, and the above process was repeated until the eighth tube.

[0074] After being mixed well on the vortex shaker, 4 μL of the bacteria solution was taken using a multichannel pipette and spotted on the treated liquid medium.

[0075] The medium was cultured in a 30°C incubator for 48 h, and when the colonies were grown, photographs were taken for observation.

[0076] (4) Test results:

[0077] The growth of Saccharomyces cerevisiae in each treatment group is shown in Figure 1. It can be observed that: (a) the gradient-diluted Saccharomyces cerevisiae can grow normally in the emulsifier control group, showing pink color without obvious inhibition effect. (b) n-Hexanol, n-Hexadecanol, n-Octadecanol, n-Eicosanol, and n-Docosanol treatment groups have no obvious inhibition effect on Saccharomyces cerevisiae at a low concentration of 0.04% (V / V). (c) Saccharomyces cerevisiae shows a slight inhibition phenomenon in n-Octadecanol and n-Tetradecanol medium, and its color is mostly light pink or milky white. (d) n-Decanol and n-Dodecanol can significantly inhibit the growth of Saccharomyces cerevisiae.

[0078] From the above test results, it can be seen that at a low concentration, n-Hexanol, n-Hexadecanol, n-Octadecanol, n-Eicosanol, and n-Docosanol do not produce significant physiological toxicity to Saccharomyces cerevisiae, while n-Decanol and n-Dodecanol can significantly inhibit the growth of Saccharomyces cerevisiae, and have a strong toxic effect on Saccharomyces cerevisiae.

[0079] (B) Morphological effects of water emulsion containing higher aliphatic alcohols on the growth of Saccharomyces cerevisiae

[0080] (1) Test agents: sample C, sample D, and sample J in Table 2;

[0081] (2) Test fungus: Saccharomyces cerevisiae W303-1A;

[0082] (3) Field emission scanning electron microscope (FE-SEM) sample preparation

[0083] YPD base medium was prepared: yeast extract 10 g, tryptone 20 g, glucose 20 g, solid plus agar powder 20 g, add water to 1000 mL. 115°C, sterilize for 20 min.

[0084] Under sterile conditions, take the activated S. cerevisiae W303-1A culture dish, pick a single colony, inoculate 5 mL YPD liquid medium, 30°C, 180 rpm for 12 h, then take 1 mL of bacterial solution, add 100 mL YPD liquid medium, 30°C, 180 rpm incubator, culture for 12 h.

[0085] Add sample C, sample D and sample J to the bacterial solution, each diluted 2500 times, i.e. 0.04% (V / V), continue to culture for 6 h.

[0086] Use a pipette to take 1 mL of bacterial solution into a 1.5 mL centrifuge tube, 2000 g, 4°C, centrifuge for 3 min, discard the supernatant.

[0087] Fixation: In the sample centrifuge tube, add 1.5 mL of 2.5% glutaraldehyde, fix at 4°C for 24 h or more.

[0088] Rinse: The fixed sample is rinsed with PBS (phosphate buffer) for 3 times, 15 min each time, to remove the glutaraldehyde on the surface of the sample.

[0089] Dehydration: Use 50%, 70%, 80%, 90% gradient ethanol to dehydrate the mycelium / spores, each time for 15 min; then 100% ethanol for 10 min.

[0090] Replacement: 50%, 75%, 90% gradient tert-butyl alcohol for replacement (tert-butyl alcohol is diluted with ethanol by volume), each time for 10 min; then 100% tert-butyl alcohol for replacement for 10 min.

[0091] Finally freeze-dried, gold-coated and observed the changes in the ultrastructure of the fungal mycelium surface by scanning electron microscopy.

[0092] (4) SEM electron microscope image analysis

[0093] The changes of the morphology of the Saccharomyces cerevisiae cells before and after the treatment were observed by field emission scanning electron microscopy (FE-SEM). It can be observed that: (a) as shown in Figure 2, the surface of the Saccharomyces cerevisiae cells treated by the control emulsifier (sample J) was complete, regular ellipsoidal or spherical, and the cell surface was not obviously damaged, and the cells could normally bud and reproduce. (b) as shown in Figure 3, after being treated by n-decanol (sample C) with a concentration of 0.04% (V / V) for 6 hours, the cell morphology changed more seriously, the cell wall was damaged, the cells were also severely contracted, and the whole surface of the Saccharomyces cerevisiae cells was obviously shrunk, and most of them could not normally bud. (c) as shown in Figure 4, after being treated by n-dodecanol (sample D) with a concentration of 0.04% (V / V) for 6 hours, most of the Saccharomyces cerevisiae cells were damaged and appeared concave.

[0094] As can be seen from the above changes of the morphology of the Saccharomyces cerevisiae cells, the treatment of the high-grade fatty alcohol water emulsion can cause damage to the cell wall of the Saccharomyces cerevisiae, and seriously affect the growth of the Saccharomyces cerevisiae.

[0095] (Three) Exploring the physiological and biochemical effects under stress by taking Saccharomyces cerevisiae as a substitute host

[0096] (1) Test reagent: sample C, sample D, sample J in Table 2

[0097] (2) Test fungus:

[0098] Saccharomyces cerevisiae W303-1A

[0099] (3) Preparation of Saccharomyces cerevisiae bacterial suspension

[0100] The low-temperature preserved Saccharomyces cerevisiae was inoculated on YPD solid medium and cultured at 30°C for 48 hours.

[0101] A single colony of Saccharomyces cerevisiae was picked and inoculated in 5 mL of YPD liquid medium, and cultured at 30°C, 180 rpm for 12 hours or so.

[0102] 1 mL of bacterial solution was taken and added to 100 mL of YPD liquid medium, and cultured on a 30°C, 180 rpm incubator for 13-16 hours to reach the logarithmic phase of the Saccharomyces cerevisiae cell growth, and the OD600 value was about 0.8.

[0103] The cells were collected in the culture system, centrifuged at 4°C, 4000g for 15 minutes, and the Saccharomyces cerevisiae cells were collected.

[0104] The cells were rinsed twice with 0.01M PBS, and the same volume of PBS was added before resuspension. The bacterial solution meeting the culture time was prepared into a bacterial suspension.

[0105] The liquid medium containing 0.04% (V / V) of n-decanol and n-dodecanol was added with 1 mL of the yeast cell suspension cultured to the logarithmic phase, so that the total volume of the culture system was 30 mL. The culture system was cultured at 30°C and 180 rpm in the dark, and the culture system added with the same concentration of emulsifier was used as a control. The yeast cells were collected at 6 h after the start of the culture for analysis.

[0106] (4) Detection of cell permeability

[0107] (a) Cell permeability detection method:

[0108] The n-decanol, n-dodecanol and emulsifier were added to the yeast cell suspension, i.e., the n-decanol, n-dodecanol and emulsifier at a concentration of 0.04% (V / V), and the culture was carried out at 30°C for 0 min, 5 min, 10 min and 15 min. After the specified time was reached, the supernatant of each bacterial solution was collected by centrifugation at 4°C and 4000 g for 5 min. The absorbance of the supernatant at 260 nm and 280 nm was measured by a multifunctional enzyme marker. The nucleic acid and protein concentrations were calculated according to the following formula, respectively.

[0109] DNA concentration (μg / mL) = 62.9 x A260 - 36.0 x A280;

[0110] Protein concentration (μg / mL) = 1552 x A280 - 757.3 x A260.

[0111] (b) Cell permeability detection results:

[0112] When the cell wall of the eukaryotic cell is damaged, the macromolecular contents in the cell, such as nucleic acid and protein, will leak into the culture medium, and the absorbance at 260 nm and 280 nm is widely used as an indicator for detecting the leakage of nucleic acid and protein from the cell into the outside of the cell. As shown in Tables 3 and 4, there is no obvious change in the extracellular DNA and protein of the yeast cells after treatment with the emulsifier, indicating that the emulsifier as a control has no significant effect on the permeability of the cell wall and cell membrane of the yeast cells under the treatment in the present experiment, while the leakage of DNA and protein from the yeast cells is significantly increased after treatment with n-decanol and n-dodecanol, which also indicates that the permeability of the cell wall and cell membrane of the yeast cells is increased after treatment. Moreover, the concentration of the extracellular leakage is positively correlated with the treatment time.

[0113] Table 3: Extracellular DNA concentration of the yeast cells (μg / mL) under each treatment

[0114] Table 4: Extracellular protein concentration of the yeast cells (μg / mL) under each treatment

[0115] (4) Detection of cell integrity

[0116] (a) Cell integrity detection method:

[0117] The cells were collected in the above culture system, centrifuged at 4000g for 10 min at 4°C, and the S. cerevisiae cells were collected.

[0118] After the yeast cells were collected, they were rinsed twice with PBS, and each time after rinsing, the cells were centrifuged at 4000g for 5 min, and the supernatant was discarded.

[0119] An equal volume of PBS was added to prepare a bacterial suspension, and the suspension was divided into 1 mL to 1.5 mL EP tubes.

[0120] The mycelium was stained with 100 μL of propidium iodide (PI) solution (20 μg / mL).

[0121] After incubation at 37°C in the dark for 15 min, the cells were rinsed with PBS solution 3 times.

[0122] After mixing with 1 mL of PBS, 500 μL was taken into a flow cytometry tube for flow cytometry detection. 10,000 cells were counted. The experiment was repeated 3 times.

[0123] (b) Cell integrity detection results:

[0124] Propidium iodide (PI) is a nucleic acid dye that is repelled by cells with intact cell membrane structures and cannot penetrate them. However, when the permeability of the cell membrane changes or the structure of the cell membrane is damaged, PI dye can easily enter these cells and tightly bind to DNA or RNA inside the cells, emitting red fluorescence.

[0125] The single-parameter histogram of flow cytometry detection under each sample treatment is shown in Figure 5. Under emulsifier treatment, the barrier function of the cell wall and cell membrane of S. cerevisiae cells is normal, and PI cannot enter the cells to stain. After n-decanol and n-dodecanol treatment, the barrier function is destroyed, and the ratio of stained cells increases sharply to 97.59% and 91.30%, respectively. The more dye that enters the cell due to changes in cell membrane permeability, the greater the fluorescence intensity. As shown in Table 5, the fluorescence intensity of S. cerevisiae treated with n-decanol and n-dodecanol is much greater than that of S. cerevisiae treated with emulsifier, indicating that n-decanol and n-dodecanol can significantly affect the cell membrane permeability of S. cerevisiae.

[0126] Table 5 PI average fluorescence intensity of S. cerevisiae cells under each treatment

[0127] (5) Detection of cellular reactive oxygen species (ROS)

[0128] (a) Cell reactive oxygen species detection method:

[0129] After 0, 3, 6 h of culture, the cells were collected in the above culture system, centrifuged at 4000 g for 10 min at 4 °C, and the S. cerevisiae cells were collected. After the yeast cells were collected, they were rinsed twice with PBS, centrifuged at 4000 g for 5 min after each rinse, and the supernatant was discarded. An equal volume of PBS was added to prepare a bacterial suspension, which was divided into 1 mL or 1.5 mL EP tubes. 10 μL of active oxygen positive control reagent Rosup was added, mixed well, and incubated at 37 °C for 20 min. The control group was added with 200 μL of active oxygen dye DCFH-DA, mixed well, and incubated at 37 °C for 20 min in the dark. Rinsed with PBS for 3 times.

[0130] After mixing with 1 mL of PBS, 500 μL was taken into a flow cytometry tube, and flow cytometry was used for detection. 10,000 cells were counted. The test was repeated 3 times.

[0131] (b) Cell active oxygen detection

[0132] As a key intermediate of aerobic metabolism, ROS levels rise sharply when cells are exposed to external stress, which is an important feature of cell stress response. Appropriate ROS can regulate cell growth, proliferation and differentiation, but excessive ROS can seriously threaten cell structure, especially by oxidizing cell membrane unsaturated fatty acids, inducing membrane peroxidation, damaging cell membrane integrity and function. At the same time, excessive ROS can also damage mitochondrial function, weaken mitochondrial membrane potential, and affect energy metabolism.

[0133] The experiment used cells without dye staining as negative control and Rosup-stained cells as positive control. The single-parameter histogram of flow cytometry detection of each sample is shown in Figure 6. The number of positive cells stained under emulsifier treatment was low, which was 1.86%; while the number of positive cells stained under n-decanol and n-dodecanol treatment was larger, which was 11.92% and 11.8%, respectively. From the change of fluorescence intensity of each treatment in Table 6, it was found that the fluorescence intensity of S. cerevisiae treated with n-decanol and n-dodecanol showed significant difference compared with the emulsifier control.

[0134] Table 6 ROS average fluorescence intensity of S. cerevisiae cells under each treatment

[0135] (6) Detection of MDA (malondialdehyde) content in S. cerevisiae

[0136] (a) Detection of MDA content in S. cerevisiae

[0137] The cells were collected in the above culture system, centrifuged at 4°C and 4000g for 10 min, and the S. cerevisiae cells were collected. After the yeast cells were collected, they were washed twice with PBS, and centrifuged at 4000g for 5 min after each washing. An equal volume of PBS was added and mixed, and the cells were broken by ultrasonic treatment (200W power, 3s ultrasonic treatment, 10s interval, repeated 30 times). The supernatant was collected by centrifugation at 4000g for 5 min, and the determination was performed according to the kit instructions.

[0138] (b) MDA content detection results in S. cerevisiae

[0139] When the level of reactive oxygen species (ROS) in cells abnormally increases, strong oxidative molecules will attack the cell membrane and protein structure, triggering the peroxidation reaction 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 not only directly reflects the peroxidation level of cell membrane lipids, but also has the ability to interact with various components in the cell, further amplifying the destructive effect of ROS and exacerbating the damage to the cell membrane.

[0140] As can be seen from Table 7, as the stress time increases, the content of MDA in the cells also increases. Compared with the emulsifier control group, the content of MDA in S. cerevisiae treated with n-decanol and n-dodecanol increased significantly, but the content of MDA in S. cerevisiae treated with n-decanol was higher, and the degree of membrane lipid oxidation was higher, which indicated that n-decanol had a stronger virulence effect on S. cerevisiae.

[0141] Table 7 MDA content in S. cerevisiae cells under different treatments (nmol / mL)

[0142] (7) Detection of mitochondrial membrane potential of cells

[0143] (a) Method for detecting mitochondrial membrane potential of cells

[0144] The cells were collected in the above culture system, centrifuged at 4000 g for 10 min at 4°C, and the S. cerevisiae cells were collected. After the yeast cells were collected, they were washed twice with PBS, and each time after washing, they were centrifuged at 4000 g for 5 min. An equal volume of PBS was added to prepare a bacterial suspension, and 1 mL to 2 mL of the bacterial suspension was dispensed into an EP tube. In the sample of the positive control group, 500 μL of CCCP dye with a dilution concentration of 10 μM was added, and the sample was incubated at 37°C for 25 min. After the pretreated cells were added with 500 μL of JC-1 staining solution, they were mixed and incubated at 37°C for 15 min in the dark. After 500 μL of JC-1 staining buffer was added, the cells were centrifuged at 4000 g for 5 min, and the supernatant was discarded. After 1 mL of JC-1 staining buffer was added, the cells were centrifuged at 4000 g for 5 min, and the supernatant was discarded. The cells were rinsed twice with the JC-1 staining buffer. After the supernatant was discarded, 1 mL of the JC-1 staining buffer was added to the cells, which were mixed and detected. 10 μL of the yeast suspension was taken to a glass slide, and the prepared sample was observed and photographed under a fluorescence microscope.

[0145] (b) Cell mitochondrial membrane potential detection results

[0146] Mitochondria, as the key place for the synthesis of adenosine triphosphate (ATP) in cells, the stable maintenance of the membrane potential is crucial for the energy conversion process. The decrease of mitochondrial membrane potential is widely recognized as one of the early biological markers of the apoptosis process.

[0147] In the experiment, the cells treated with emulsifiers were used as negative controls, and the cells stained with CCCP were used as positive controls. As can be seen from Tables 8A, 8B and 8C, compared with the emulsifier treatment group, the green fluorescence of the n-decanol and n-dodecanol treatment groups was significantly improved, showing a significant difference, and the RFI further illustrated that n-decanol and n-dodecanol had significant toxicity to S. cerevisiae, which could cause a sharp decline in the mitochondrial transmembrane potential, and thus rapidly weaken the mitochondrial activity.

[0148] Table 8A Fluorescence intensity of mitochondrial transmembrane potential of S. cerevisiae cells under different treatments (red light)

[0149] Table 8B Fluorescence intensity of mitochondrial transmembrane potential of S. cerevisiae cells under different treatments (green light)

[0150] Table 8C Fluorescence intensity of mitochondrial transmembrane potential of S. cerevisiae cells under different treatments (RFI)

[0151] (IV) Transcriptome analysis of S. cerevisiae growth with water emulsions containing higher fatty alcohols

[0152] (1) Test agents: sample D and sample J in Table 2

[0153] (2) Test fungi:

[0154] Saccharomyces cerevisiae W303-1A

[0155] (3) Preparation of transcriptome samples

[0156] Each bacterial solution (OD600 was about 0.8) was treated with n-dodecanol (sample D) at a concentration of 0.04% (V / V) for 5 min and 1 h at 30°C, and the emulsifier (sample J) treatment group was used as the control group. The control group and the experimental group were set up in three biological replicates. RNA extraction and RNA-Seq sequencing were commissioned to Wuhan Maiwei Metabolic Biology Technology Co., Ltd.

[0157] (4) Transcriptome analysis and results

[0158] (a) Screening of differential genes

[0159] For the sequencing results, DESeq2 was used to screen the differential genes, with the screening conditions being |log2FoldChange|≥1 and FDR<0.05, to obtain the differentially expressed genes.

[0160] As shown in the volcano plot of the differentially expressed genes (Figures 7 and 8), compared with the control group, 4333 differential genes were produced after n-dodecanol treatment of Saccharomyces cerevisiae for 5 min, including 877 up-regulated genes and 620 down-regulated genes;

[0161] Compared with the control group, 4115 differential genes were produced after n-dodecanol treatment of Saccharomyces cerevisiae for 1 h, including 1072 up-regulated genes and 650 down-regulated genes.

[0162] (b) GO enrichment analysis of differentially expressed genes

[0163] After dodecanol treatment for 5 min, the differentially expressed genes were mainly distributed in 15 biological processes, 15 cell compositions and 15 molecular functions. Figure 9 shows the significant enrichment of these differentially expressed genes in the form of scatter plots, and the most significant 20 GO-Term are displayed. The results show that GO-Term includes 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.

[0164] After dodecanol treatment for 1 h, the differentially expressed genes were mainly distributed in 15 biological processes, 2 cell compositions and 15 molecular functions. Figure 10 shows the significant enrichment of these differentially expressed genes in the form of scatter plots, and the most significant 20 GO-Term are displayed. The results show that the differentially expressed genes are mainly mitochondrial protein-containing complex, mitochondrial inner membrane, organellar inner membrane, mitochondrial membrane, mitochondrial matrix, mitochondrial ribosome, etc.

[0165] The GO enrichment analysis results show that the mitochondria and organelles of S. cerevisiae are affected by dodecanol stress for 5 min and 1 h. This indicates that dodecanol treatment may cause greater damage to the mitochondria or other organelles of S. cerevisiae.

[0166] (c) KEGG enrichment analysis of differentially expressed genes

[0167] Figure 11 shows that after KEGG annotation of the differential genes of S. cerevisiae under dodecanol stress for 5 min, it was found that the differential genes were mainly concentrated in 75 metabolic pathways, 22 genetic information processing pathways, 8 cell process pathways, etc. In addition, the 5 pathways with the most differential expression genes were ribosome (ko03010) related to genetic information, yeast MAPK signaling pathway (ko04011), amino acid synthesis pathway (ko01230), yeast-autophagy pathway (ko04138), and oxidative phosphorylation (ko00190).

[0168] Figure 12 shows that after KEGG annotation of the differential genes of S. cerevisiae under dodecanol stress for 1 h, it was found that the differential genes were mainly concentrated in 77 metabolic pathways, 22 genetic information processing pathways, 8 cell process pathways, etc. In addition, among the top 20 pathways with the most significant differential expression genes, the 5 pathways with the most differential expression genes were metabolic pathways (ko01100), secondary metabolite biosynthesis pathways (ko01110), amino acid synthesis pathways (ko01230), yeast MAPK signaling pathways (ko04011), and oxidative phosphorylation (ko00190).

[0169] From the KEGG enrichment analysis, both groups of treatment groups with different lengths of time can affect similar pathways, such as yeast MAPK signaling pathway (ko04011) and oxidative phosphorylation (ko00190), indicating that as the time of dodecanol stress increases, there is a continuous impact on the MAPK signaling, amino acid synthesis, and oxidative phosphorylation functions of S. cerevisiae. The MAPK signaling pathway in S. cerevisiae cells is related to spore differentiation, hyphal formation and embedded growth, high osmotic glycerol formation, cell wall integrity, and cell binding processes. Under the stress of dodecanol, HKR1, WSC3, GPD2, TUS1, and other cell wall integrity signaling enzyme-related genes were down-regulated, affecting the signaling and maintenance of cell wall integrity. Oxidative phosphorylation occurs on the inner membrane of the mitochondria of eukaryotic cells and is an important pathway for ATP synthesis in cells. The impact of dodecanol on these pathways directly affects the normal function of the mitochondria of S. cerevisiae cells.

[0170] (d) Important pathway and gene analysis

[0171] After 1h of dodecanol stress, 36 genes related to oxidative phosphorylation pathway were significantly up-regulated compared with the control group. The top ten up-regulated genes in the oxidative phosphorylation pathway were determined and analyzed. The results showed that dodecanol could significantly up-regulate the transcription level of related genes in mitochondria, and the transcription level was high. These genes include succinate dehydrogenase (SDH), cytochrome C (CYC), cytochrome C oxidase (COX), mitochondrial matrix protein NDUS7, type II NADH dehydrogenase (NDH2), etc.

[0172] Succinate dehydrogenase (SDH) is a key complex enzyme in the tricarboxylic acid cycle and the respiratory chain of electron transfer in mitochondria. SDHx gene mutation leads to mitochondrial dysfunction, which in turn activates the pseudo-hypoxic signaling pathway. SDHA and SDHB in succinate dehydrogenase are significantly up-regulated, leading to mitochondrial dysfunction. At the same time, cytochrome C (CYC) is also released in large quantities from mitochondria, aggravating the electron transfer disorder of the mitochondrial respiratory chain and increasing the production of ROS, promoting cell apoptosis.

[0173] As can be seen from the sequencing analysis results, dodecanol stress can significantly up-regulate the transcription level of related genes in mitochondria, and the transcription level is high, indicating that dodecanol has caused damage to the mitochondria of the cell, which is consistent with the sharp decline in mitochondrial transmembrane potential observed in the physiological and biochemical experiments.

[0174] From the above test results of the growth, physiological and biochemical effects, and transcription sequencing analysis of the model organism Saccharomyces cerevisiae after high-grade fatty alcohol stress, it can be seen that high-grade fatty alcohols dodecanol and decanol can directly damage the cell wall and mitochondria of fungal cells, leading to cell structure damage, endogenous substance outflow, and mitochondrial dysfunction, causing growth inhibition, damage, and death of fungal cells. This fully demonstrates that the preparation containing high-grade fatty alcohols has a bacteriostatic and fungicidal effect on fungi.

[0175] Example 5: Effect of water emulsion containing high-grade fatty alcohols on the growth of Fusarium oxysporum

[0176] (1) Test agent: water emulsion prepared according to Table 2

[0177] (2) Test fungi:

[0178] Fusarium oxysporum f.sp.cubense (FOC009)

[0179] (3) Sample treatment:

[0180] PDA basic medium: 200g potato, washed and peeled, cut into small pieces, add water 1000mL, boil for 30min; filter with gauze, add water to make up to 1000mL, then add 20g glucose, and add 20g agar to make it fully dissolved, sterilize with high-pressure sterilization pot at 115℃ for 20min, and then use.

[0181] The different components of the liquid medicine were taken with a pipette and added to the PDA medium which had not solidified after sterilization, diluted 5000 times, 2500 times, 1000 times, 500 times and 100 times, i.e. the concentration was 0.02%, 0.04%, 0.1%, 0.2% and 1% (V / V) respectively, and then shaken well. The culture medium containing the medicine was poured into 90mm culture dishes, about 15mL per dish.

[0182] Under sterile conditions, a punch with a diameter of 8mm was used to take the fungus cake activated in the PDA medium, the fungus cake was required to grow uniformly, the mycelium surface of the fungus cake was placed downward, and the fungus cake was inoculated into the central position of the solidified PDA medium plate and placed in a 26℃ artificial climate box for culture. After 5d of culture, the blank control was treated with water, the growth index of the colonies was observed and determined, and the inhibitory effect of the high-fat film and different components on different fungi was determined.

[0183] (4) Data collection and processing:

[0184] The activated fungus cake was inoculated on the PDA solid medium, and cultured in a 26℃ incubator for about 5d, and the mycelium of the control group covered more than 3 / 4 of the plate.

[0185] The mycelial growth was observed and recorded, and the colony diameter was measured by cross method after 5d, and the mycelial growth inhibition rate was calculated according to the following formula:

[0186] Mycelial growth inhibition rate = (colony growth diameter of control group - colony growth diameter of treatment group) / colony growth diameter of control group x 100%.

[0187] The effects of various drugs on the culture traits of the pathogenic fungi were compared by observing and recording the colony morphology, whether the colony edge was neat, whether the colony surface was flat, etc. after culture at various concentrations.

[0188] (5) Effects of different higher aliphatic alcohols on the growth of Fusarium oxysporum:

[0189] Microscopic observation showed that, as shown in Figure 13, under the same concentration treatment, compared with the emulsifier control group, n-decanol and n-dodecanol had significant fungal growth inhibition ability, the colony diameter of FOC009 treated with n-decanol and n-dodecanol was small, round and milky white, and the mycelium was dense. Under the treatment of other drugs, the colony was round, with a large amount of white aerial mycelium, the edge was irregularly serrated, and it was opaque.

[0190] The growth rate of Fusarium oxysporum f. sp. cubense (FOC009) after treatment with different higher aliphatic alcohols is shown in Table 9. It can be seen that the growth rate of FOC009 after treatment with different higher aliphatic alcohols at the same concentration is low, being 3.28 mm / d and 8.70 mm / d for n-decanol and n-dodecanol, respectively. The growth rate of the colonies of FOC009 treated with emulsifier, n-hexanol, n-octanol, n-tetradecanol, n-hexadecanol, n-octadecanol, n-eicosanol and n-docosanol is similar, being more than 10.00 mm / d.

[0191] The inhibition rate of higher aliphatic alcohols on the growth of FOC009 is shown in Table 10. The inhibition effect of n-decanol, n-dodecanol and n-tetradecanol is better, the inhibition rate being 76.40%, 38.92% and 29.35%, respectively. The inhibition effect of other higher aliphatic alcohols is general.

[0192] Table 9 Growth rate of FOC009 treated with higher aliphatic alcohols

[0193] Table 10 Inhibition rate of higher aliphatic alcohols on the growth of FOC009

[0194] Example 6: Effect of water emulsion containing higher aliphatic alcohols on the growth of plant pathogenic fungi and indoor toxicity analysis

[0195] (1) Test agent: water emulsion prepared according to Table 11

[0196] Table 11 Sample number

[0197] High-fat film is the water emulsion of sample S, containing n-dodecanol and n-hexadecanol, i.e. the water emulsion containing higher aliphatic alcohols provided in Example 1, and the same applies hereinafter.

[0198] (2) Test fungi:

[0199] Fusarium graminearum;

[0200] Fusarium oxysporum f. sp. cubense (FOC009);

[0201] Colletotrichum gloeosporioides.

[0202] (3) Sample treatment:

[0203] PDA basic medium: 200g potato, washed and peeled, cut into small pieces, add water 1000mL, boil for 30min; filter with gauze, add water to make up to 1000mL, then add 20g glucose, and add 20g agar to make it fully dissolved, sterilize with high-pressure sterilization pot at 115℃ for 20min, and then use.

[0204] The different components of the liquid medicine were taken with a pipette and added to the PDA medium which had not yet solidified after sterilization, diluted 5000 times, 2500 times, 1000 times, 500 times and 100 times, i.e. the concentration was 0.02%, 0.04%, 0.1%, 0.2% and 1% (V / V) respectively, and then shaken thoroughly. The culture medium containing the medicine was poured into 90mm culture dishes, about 15mL per dish.

[0205] Under sterile conditions, a punch with a diameter of 8mm was used to take the fungus cake activated in the PDA medium, the fungus cake was required to grow uniformly, the mycelium surface of the fungus cake was placed downward, and the fungus cake was inoculated into the central position of the solidified PDA medium plate and placed in a 26℃ artificial climate box for culture. After 5d of culture, the blank control was treated with water, the growth of the colonies was observed and measured, and the inhibitory effect of the high-fat film and different components on different fungi was determined.

[0206] (4) Data collection and processing:

[0207] The activated fungus cake was inoculated on the PDA solid medium, and cultured in a 26℃ incubator for about 5d, and the mycelium of the control group covered more than 3 / 4 of the plate.

[0208] The growth of the mycelium was observed and recorded, and the colony diameter was measured by the cross method after 5d, and the mycelial growth inhibition rate was calculated according to the following formula:

[0209] Mycelial growth inhibition rate = (colony growth diameter of the control group - colony growth diameter of the treatment group) / colony growth diameter of the control group x 100%.

[0210] Toxicity regression equation and effective concentration calculation. The toxicity of each fungicide to the mycelium of the fungus was expressed by the mycelial growth inhibition rate. The mycelial growth inhibition rate was converted into an inhibition probability value (y), and the concentration of the fungicide was converted into the logarithm of the concentration (x), and toxicity regression analysis was performed to calculate the EC50 value and EC90 value of each test pathogenic fungus and the correlation coefficient R value of the regression equation.

[0211] The influence of each fungicide on the culture characteristics of the pathogen was compared by observing and recording the colony morphology, whether the colony edge was neat, whether the colony surface was flat, etc. after culture at each concentration.

[0212] (5) Influence of different higher fatty alcohols on the growth of Fusarium graminearum, Fusarium oxysporum and Colletotrichum gloeosporioides:

[0213] (a) Effect of different higher aliphatic alcohol components on the growth of Fusarium graminearum colony

[0214] Microscopic observation showed that, as shown in Figure 14, under the treatment of each component at a dilution concentration of 1% (V / V), the colony diameter of Fusarium graminearum treated with lauryl alcohol was the smallest, and the colony was round, light pink, the center was darker, and the edge was smooth. The colony treated with high-fat film (GZM) was similar to that treated with lauryl alcohol, but the colony growth diameter was slightly larger, the colony was round, the center was deep rose, there were white aerial hyphae around, and the edge was irregularly serrated and slightly transparent. Under the treatment of each component sample at a dilution concentration of 0.2%, the colony diameters treated with high-fat film and lauryl alcohol were similar, the center was round orange red, and the edge was light red; under the treatment of the rest of the component samples, the center was round light red or dark red, there were a large number of hyphae around, mainly with lower layer pink hyphae and upper layer white or yellow hyphae, the edge was rodent-like and opaque.

[0215] The results showed that, under the treatment of each component sample at a dilution concentration of 1%, lauryl alcohol and high-fat film had good inhibitory effect on the mycelial growth of Fusarium graminearum. The emulsifier did not have obvious inhibitory effect on the mycelial growth of Fusarium graminearum at different concentrations.

[0216] (b) Effect of different higher aliphatic alcohol components on the growth of Fusarium oxysporum colony

[0217] Microscopic observation showed that, as shown in Figure 15, under the treatment of each component sample at a dilution concentration of 1%, the colony diameter of Fusarium oxysporum treated with lauryl alcohol was the smallest, and the colony was round, with short and dense hyphae, and the edge was uneven and opaque. The colony treated with high-fat film (GZM) was similar to that treated with lauryl alcohol, but the edge of the colony was smoother. Under the treatment of the rest of the samples, the colony was round, with a large number of white aerial hyphae, and the edge was regular serrated and slightly transparent.

[0218] The results showed that, under the treatment of each component sample at a dilution concentration of 1%, lauryl alcohol and high-fat film had good inhibitory effect on Fusarium oxysporum, and the inhibitory effects were similar. Under the treatment of each component at a dilution concentration of 0.02% and 0.04%, high-fat film and lauryl alcohol still had certain inhibitory effect on Fusarium oxysporum.

[0219] (c) Effect of different higher aliphatic alcohol components on the growth of Colletotrichum gloeosporioides colony

[0220] Microscopic observation showed that, as shown in Figure 16, under the treatment of each component at a dilution concentration of 1%, the colony diameter of the G. cingulata treated with lauryl alcohol was the smallest, which was round, milky white, gray in the center, irregular jagged at the edge, and opaque. The colony treated with GZM was similar to that treated with lauryl alcohol. Under the treatment of the remaining components, the colony was round, yellow-brown in the center, with a large amount of white aerial hyphae, regular jagged at the edge, and opaque.

[0221] The results showed that, under the treatment of each component at a dilution concentration of 1%, GZM and lauryl alcohol had good inhibitory effect on G. cingulata, and the inhibitory effect was similar.

[0222] (d) Inhibitory activity of different higher aliphatic alcohol components on the colony growth of F. graminearum

[0223] The growth inhibition rate of F. graminearum mycelium under different treatments is shown in Table 12. Different concentrations of different higher aliphatic alcohol component samples had different effects on F. graminearum during cultivation. Under the treatment of 0.02% low concentration, the growth of F. graminearum was not inhibited. With the increase of concentration, GZM, lauryl alcohol and lauryl alcohol all had a certain inhibitory effect on the colony growth of F. graminearum, and showed a positive trend.

[0224] Table 12 Growth inhibition rate of F. graminearum mycelium under different treatments

[0225] (e) Inhibitory activity of different higher aliphatic alcohol components on the colony growth of F. graminearum

[0226] The growth inhibition rate of F. graminearum mycelium under different treatments is shown in Table 12. Different concentrations of different higher aliphatic alcohol component samples had different effects on F. graminearum during cultivation. Under the treatment of 0.02% low concentration, the growth of F. graminearum was not inhibited. With the increase of concentration, GZM, lauryl alcohol and lauryl alcohol all had a certain inhibitory effect on the colony growth of F. graminearum, and showed a positive trend.

[0227] Table 13 Growth inhibition rate of F. graminearum mycelium under different treatments

[0228] (f) Inhibitory activity of different higher aliphatic alcohol components on the colony growth of G. cingulata

[0229] The growth inhibition rates of the mycelium of Colletotrichum gloeosporioides under different treatments are shown in Table 14. Different concentrations of different high-grade fatty alcohol component samples have different effects on the production of Colletotrichum gloeosporioides during the culture process. Under the treatment of 0.02% low concentration, high lipid membrane has no inhibitory effect on the growth of Colletotrichum gloeosporioides. With the increase of concentration, high lipid membrane, n-decanol and n-dodecanol all have a certain inhibitory effect on the growth of Colletotrichum gloeosporioides, and show a positive trend.

[0230] Table 14 Growth inhibition rate of mycelium of Colletotrichum gloeosporioides under different treatments

[0231] (7) Toxicity analysis of different high-grade fatty alcohol components on test fungi

[0232] The toxicity regression analysis of different high-grade fatty alcohol components on the indoor toxicity of 3 common plant pathogenic fungi is shown in Table 15. In addition to the unstable bacteriostatic effect of emulsifiers on the test plant pathogenic fungi, different high-grade fatty alcohol components all have a certain inhibitory effect on the 3 test fungi. Among them, high lipid membrane, n-decanol and n-dodecanol have good bacteriostatic activity on the 3 test fungi. The EC50 of high lipid membrane on Fusarium graminearum, Fusarium oxysporum and Colletotrichum gloeosporioides is 0.5197%, 0.3676% and 0.7115% respectively. The EC50 of n-decanol on Fusarium graminearum, Fusarium oxysporum and Colletotrichum gloeosporioides is 0.5657%, 0.0837% and 0.0656% respectively. The EC50 of n-dodecanol on Fusarium graminearum, Fusarium oxysporum and Colletotrichum gloeosporioides is 0.4193%, 0.2802% and 0.7445% respectively.

[0233] Table 15 Indoor toxicity of high lipid membrane and its main components on 3 common plant pathogenic fungi

[0234] Example 7: Effect of water emulsion containing high-grade fatty alcohol on the growth of Peronophythora litchii and indoor toxicity analysis

[0235] (1) Test agent: water emulsion prepared according to the present application as listed in Table 16

[0236] Table 16 Sample No.

[0237] High lipid membrane is the sample S water emulsion, containing n-dodecanol and n-hexadecanol, two high-grade fatty alcohols.

[0238] (2) Test oomycetes:

[0239] Peronophythora litchii.

[0240] (3) Sample treatment:

[0241] Carrot agar medium: 200g carrot, washed, peeled, cut into small pieces, squeezed juice, filtered with gauze, add water to make up to 1000mL, add 20g agar, dissolve thoroughly, sterilize with high pressure sterilization pot at 115℃ for 20min, and use.

[0242] The different components of the liquid medicine were taken with a pipette and added to the non-solidified medium after sterilization, diluted 5000 times, 2500 times, 1000 times, 500 times and 100 times, i.e. the concentration was 0.02%, 0.04%, 0.1%, 0.2% and 1% (V / V) respectively, and then shaken thoroughly. The culture medium containing the medicine was poured into 90mm culture dishes, about 15mL per dish.

[0243] Under sterile conditions, the oomycete fungus cake activated in the carrot agar medium was punched with a puncher with a diameter of 8mm, the fungus cake was required to grow uniformly, the mycelium surface of the fungus cake was placed downward, and the fungus cake was inoculated into the central position of the solidified PDA medium plate and placed in a 26℃ artificial climate box for culture. After 7d of culture, the blank control was treated with water, the colony growth indicators were observed and determined, and the inhibitory effect of high-fat membranes and different components on different oomycetes was determined.

[0244] (4) Data collection and processing:

[0245] The activated oomycete fungus cake was inoculated in carrot agar medium and cultured in a 26℃ incubator for about 7d, and the mycelium of the control group covered more than 3 / 4 of the plate.

[0246] The mycelium growth was observed and recorded, the colony diameter was measured by cross method after 7d, and the mycelium growth inhibition rate was calculated according to the following formula:

[0247] Mycelium growth inhibition rate = (colony growth diameter of control group - colony growth diameter of treatment group) / colony growth diameter of control group x 100%.

[0248] Toxicity regression equation and effective concentration calculation. The toxicity of each pesticide to oomycete mycelium was expressed by the mycelium growth inhibition rate. The mycelium growth inhibition rate was converted into inhibition probability value (y), and the concentration of the pesticide was converted into concentration logarithm (x), and toxicity regression analysis was performed to calculate the EC50 value and EC90 value of each test pathogenic fungus and the regression equation correlation coefficient R value.

[0249] The influence of each pesticide on the culture characteristics of the pathogen was compared by observing and recording the colony morphology, whether the colony edge was neat, and whether the colony surface was flat after culture at each concentration.

[0250] (5) Influence of different higher fatty alcohols on the growth of Litchi Peronophythora litchii

[0251] (a) Influence of different higher fatty alcohol components on the colony growth of Litchi Peronophythora litchii

[0252] Microscopic observation showed that, as shown in Figure 17, under the treatment of each component at a dilution concentration of 0.04%, 0.1%, 0.2% and 1% (V / V), compared with the water blank control (CK), the colony diameters of the high-fat film (sample S, GZM) and lauryl alcohol (sample D) treated L. soneriati were smaller, round, and the edges were complete and smooth. The colony shape under the treatment of high-fat film was similar to that under the treatment of lauryl alcohol. Compared with the water control, as the dilution concentration of each component sample increased, the colony diameters of the high-fat film and lauryl alcohol treated L. soneriati became smaller, indicating that the inhibitory effect on the mycelial growth of L. soneriati was stronger and stronger.

[0253] (b) Inhibition of L. soneriati colony by different high-grade fatty alcohol components

[0254] The growth inhibition rate of L. soneriati mycelium under different treatments is shown in Table 17. Different concentrations of lauryl alcohol and high-fat film had different effects on L. soneriati during the culture process. Different concentrations of lauryl alcohol and high-fat film had good inhibitory effect on the mycelial growth of L. soneriati, and the inhibitory effect increased with the increase of the concentration of the preparation, showing a positive trend.

[0255] Table 17 Growth inhibition rate of L. soneriati mycelium under different treatments

[0256] (6) Virulence determination of high-fat film and main component lauryl alcohol on test oomycetes

[0257] The virulence regression analysis of the indoor virulence determination data of high-fat film and lauryl alcohol on L. soneriati is shown in Table 18. High-fat film and lauryl alcohol had a certain inhibitory effect on L. soneriati, and the antibacterial activity was good. The EC50 concentration of high-fat film on L. soneriati was 0.0334%, and the EC50 concentration of lauryl alcohol on L. soneriati was 0.0176%.

[0258] Table 18 Indoor virulence of high-fat film and lauryl alcohol on L. soneriati

[0259] Example 8: Preparation containing high-grade fatty alcohol improves the effect of preventing and treating wheat scab

[0260] Sampling: Sample S of treatment group 1 is the water emulsion prepared in Example 1; sample C of treatment group 2 is the water emulsion prepared from test agent 3 in Example 2;

[0261] Crop: Zhongken 616 wheat

[0262] Location: Caobu Town, Rudong County, Nantong City, Jiangsu Province

[0263] Method: 20 mu of treatment group 1, 20 mu of treatment group 2, 20 mu of control group were selected in the production base for production, and the treatment group 1 and the treatment group 2 were produced as follows: the sample S was diluted 1000 times with water and then sprayed, and each time the sample dosage was 30 mL per mu, and the seedling stage, tillering stage and grain filling stage were sprayed once; the control group was sprayed with water at the same time as the treatment group 1 and the treatment group 2.

[0264] Effect comparison:

[0265] The wheat treated by the preparation emulsion containing the senior fatty alcohol of the application in the treatment group 1 and the treatment group 2 has little difference in appearance, the plant growth is dense, the leaf color is dark green and shiny, the plant is high and the stem is strong, and compared with the control group, the number of tillers is more, and the plant height, the number of leaves and the number of tillers all have obvious advantages, and the growth and appearance also have obvious advantages.

[0266] Sampling and yield results:

[0267] The yield of the treatment group 1, the treatment group 2 and the control group was measured, and 30 ear heads were randomly selected from the treatment group 1, the treatment group 2 and the control group, and the ear length, the number of grains per ear, the number of empty husks, the empty husk rate, the fusarium head blight disease rate and the thousand-grain weight were investigated and detected, and the results are shown in Table 19.

[0268] Table 19: Yield test and detection results of the wheat using the preparation containing the senior fatty alcohol of the application

[0269] Fusarium graminearum is a pathogenic fungus widely harmful to cereal crops, which can cause various important diseases and is the main pathogenic fungus of wheat fusarium head blight, affecting the growth of wheat. As can be seen from the above table, the fusarium head blight disease rate of the control group 1 and the control group 2 using the emulsion containing the senior fatty alcohol of the application is 5.50% and 5.53%, which is significantly lower than that of the control group 46.08%, the yield of the treatment group 1 is 448.8 kg per mu, the yield of the treatment group 2 is 451.6 kg per mu, and the yield of the control group is 405.4 kg per mu, the yield of the treatment group is significantly improved, and the increase is 10.71% and 11.14%, which shows that the emulsion containing the senior fatty alcohol of the application, the treatment group 1 containing dodecanol and the treatment group 2 containing decanol, all have obvious effects on the prevention and treatment of wheat fusarium head blight, which is consistent with the specific bacteriostatic and bactericidal effects of the senior fatty alcohol on fusarium graminearum shown in Table 6.

[0270] Example 9: Effect of the preparation containing the senior fatty alcohol on the prevention and treatment of diseases in peanut field

[0271] Sampling: the sample S is the emulsion prepared in Example 1

[0272] Crop: peanut shanyou 35

[0273] Location: Luoba Town, Shixing County, Shaoguan City, Guangdong Province

[0274] Method: 20 mu of treatment group and 20 mu of control group were selected for production in the production base. The treatment group was produced as follows: the sample S was diluted 50 times with water, then seed dressing was carried out, and after drying, it was sowed. The sample S was diluted 1000 times with water, then spraying was carried out, and the seedlings, flowering, podding and maturation stages were sprayed once respectively. The sample dosage was 15 mL / mu each time. At the same time of the treatment of the treatment group, the control group was sprayed with the same amount of water.

[0275] Comparison of use effect:

[0276] The peanuts treated by the emulsion in water containing the senior fatty alcohol of the application in the treatment group had no obvious disease and pest damage in the whole growth process. The plant growth of the treatment group was more dense, the leaf color was dark green and shiny, and the plant height was higher. Compared with the control group, the germination rate, growth condition and number of flower buds of the treatment group all showed obvious advantages.

[0277] Table 20: Test effect of peanuts treated by the emulsion in water containing the senior fatty alcohol of the application

[0278] The above table shows that the peanuts treated by the emulsion in water containing the senior fatty alcohol of the application had no obvious disease and pest damage and less rotten fruit, while the control group had root rot and more rotten fruit. The main pathogen of peanut root rot and rotten fruit disease is fusarium. The emulsion in water containing the senior fatty alcohol of the application has obvious bacteriostatic and bactericidal effects on fusarium, thereby reducing the occurrence of root rot and rotten fruit disease in the treatment group.

[0279] Example 10: Disease control effect of the emulsion in water containing the senior fatty alcohol on Xinhui tangerine and orange trees

[0280] Sample S is the emulsion in water prepared in Example 1;

[0281] Crop: Xinhui tangerine and orange;

[0282] Location: Shuangshui Town, Xinhui District, Jiangmen City, Guangdong Province

[0283] Method: 20 mu of treatment group and 20 mu of control group were selected for production in the production base. The treatment group was produced as follows: from the beginning of the spring fruit tree germination of tender leaves, the sample S was diluted 1000 times with water, then spraying was carried out, and the interval was 15 days. The sample dosage was 100 mL / mu each time. At the same time of the treatment of the treatment group, the control group was sprayed with the same amount of water.

[0284] Comparison of use effect:

[0285] The fruit trees treated by the emulsion in water containing the senior fatty alcohol of the application in the treatment group had strong tree vigor, dark green and shiny leaf color, no occurrence of citrus anthracnose, and less fruit drop.

[0286] Table 21 Test effect of citrus using the preparation containing higher fatty alcohol according to the application

[0287] The above table shows that the fruit trees treated with the preparation containing higher fatty alcohol according to the application have no obvious pest and disease damage and less fruit drop, while the control group has anthracnose occurrence and more fruit drop. Citrus anthracnose is caused by Glomerella cingulata, which can harm the leaves, branches, flowers and fruits of fruit trees. The preparation containing higher fatty alcohol according to the application has obvious bacteriostatic and bactericidal effects on G. cingulata, thereby reducing the occurrence of anthracnose and physiological fruit drop in the treatment group.

[0288] Example 11: Disease control effect of the preparation containing higher fatty alcohol on sugarcane

[0289] Sampling: Sample S of the treatment group 1 is the water emulsion prepared in Example 1; sample C of the treatment group 2 is the water emulsion prepared from the test agent 3 in Example 2;

[0290] Crop: sugarcane Gui Tang 44;

[0291] Location: Luobai community, Jiangzhou district, Chongzuo city, Guangxi;

[0292] Method: 50 mu of the treatment group 1, 50 mu of the treatment group 2 and 50 mu of the control group were selected for production in the production base. The treatment group 1 and the treatment group 2 were produced as follows: sample S was diluted 1000 times with water and sprayed, and spraying was performed once every 7 days, with a sample dosage of 100 mL per mu each time; at the same time of the treatment of the treatment group 1 and the treatment group 2, the control group was sprayed with the same amount of water.

[0293] Comparison of use effects:

[0294] The appearance of the sugarcane treated with the water emulsion of the preparation containing higher fatty alcohol according to the application in the treatment group 1 and the treatment group 2 has little difference, the plant leaf color is dark green and lustrous, and the growth is vigorous, while the plant leaf color of the control group is somewhat yellow, and the growth is weak.

[0295] 30 square meters were randomly selected from the treatment group 1, the treatment group 2 and the control group as investigation points to investigate the number of smut, and the number of smut was counted and statistically analyzed, as shown in Table 22.

[0296] Table 22 Disease control effect of the preparation containing higher fatty alcohol according to the application on sugarcane smut

[0297] The above table shows that the sugarcane in the treatment group 1 and the treatment group 2 treated by the preparation containing the higher aliphatic alcohol has the dark green and shiny leaf color and vigorous growth, while the leaf color of the plant in the control group is somewhat yellow and the growth is weak. The number of smut of sugarcane in the treatment group 1 and the treatment group 2 after treatment is small and the growth rate is low, and the smut of sugarcane is obviously inhibited, while the number of smut of sugarcane in the control group is more and more and the growth rate is also faster and faster. The main pathogen of the smut of sugarcane is the sugarcane smut fungus, which is the most important fungal disease in sugarcane planting and is one of the most common and most serious diseases in sugarcane cultivation. The preparation containing the higher aliphatic alcohol has the obvious fungistatic and fungicidal effects on fungi after treatment, so that the treatment group reduces the occurrence of the smut of sugarcane.

[0298] Example 12: Disease control effect of the preparation containing the higher aliphatic alcohol on cucumber in the field

[0299] Sampling: the sample S is the aqueous emulsion prepared in Example 1;

[0300] Crop: cucumber 8808;

[0301] Place: Shunzhuang Town, Daming County, Handan City, Hebei Province;

[0302] Method: 5 mu of the treatment group and 5 mu of the control group are selected for production in the production base. The treatment group is produced as follows: the sample S is diluted 100 times with water to immerse the roots of the cucumber seedlings before planting, and the sample S is diluted 800 times with water for spraying. The seedlings, flowering and harvesting stages are each sprayed once, and the sample dosage is 25 mL per mu each time. At the same time of the treatment of the treatment group, the control group is sprayed with the same amount of water.

[0303] Comparison of use effects:

[0304] The cucumber in the treatment group treated by the aqueous emulsion of the preparation containing the higher aliphatic alcohol has no obvious disease and pest damage in the whole growth process, and the plant in the treatment group has better growth, lush leaves, more flowers and more fruits, and maintains better tree vigor.

[0305] Table 23: Test effect of the cucumber treated by the preparation containing the higher aliphatic alcohol of the application

[0306] The above table shows that the cucumber treated with the preparation containing the higher fatty alcohol according to the application has good plant growth throughout the whole growth process, no obvious diseases and pests are found, and the yield is significantly improved by 96.8%, while the control group has powdery mildew and gummy stem blight throughout the whole growth process, which harms the leaves and vines of cucumber, affects the normal growth of cucumber, and seriously affects the yield; the cucumber gummy stem blight is caused by Aspergillus carterae and Mycosphaerella melonis, and the cucumber powdery mildew is caused by Sphaerotheca fuliginea and Podosphaera xanthii, which all belong to the subphylum Ascomycotina fungi. Obviously, the preparation containing the higher fatty alcohol according to the application has obvious fungistatic and fungicidal effects on fungi, and after treatment, the occurrence of cucumber powdery mildew and gummy stem blight is reduced, and the yield of cucumber is significantly improved.

[0307] Example 13: Disease control effect of the preparation containing the higher fatty alcohol in the field of long eggplant

[0308] Sampling: sample S is the aqueous emulsion prepared in Example 1;

[0309] Crop: long eggplant Heilongzun No. 2;

[0310] Location: Shujiang Town, Daming County, Handan City, Hebei Province;

[0311] Method: 2 mu of treatment group and 2 mu of control group are selected for production in the production base. The treatment group is produced as follows: the sample S is diluted 100 times with water, the eggplant seedlings are immersed in roots before planting, and the sample S is diluted with water and sprayed, and each of the seedling stage, flowering stage, fruiting stage and harvesting stage is sprayed once, and the dilution multiples of the seedling stage, flowering stage, fruiting stage and harvesting stage are 800 times, 600 times, 500 times and 500 times respectively, and the sample dosage is 60 mL per mu each time; at the same time of treating the treatment group, the control group is sprayed with the same amount of water.

[0312] Comparison of use effect:

[0313] The eggplant in the treatment group treated with the aqueous emulsion of the preparation containing the higher fatty alcohol according to the application has no obvious disease and pest damage throughout the whole growth process, the plant growth condition of the treatment group is more robust in appearance, the leaf is wider, the number of flowers and fruits per plant is more, and the appearance of the fruit is better, and the coloring is more uniform, which all show significant advantages.

[0314] Table 24: Test effect of eggplant treated with the preparation containing the higher fatty alcohol according to the application

[0315] The above table shows that eggplants treated with the preparation containing a higher fatty alcohol according to the present application have no obvious diseases and pests, the plants are strong, have more fruits, and the yield is increased by 25.9% compared with the control group. In the whole growth process, the control group is found to have eggplant gray mold and eggplant blight. The pathogen of eggplant gray mold is Botrytis cinerea, which belongs to the subphylum of Deuteromycota fungi. The pathogen of eggplant blight is Phytophthora nicotianae, which belongs to the Oomycota division of the fungal genus Phytophthora. The eggplant is invaded by these two diseases, resulting in a decrease in yield. By comparison, it can be seen that the use of the preparation containing a higher fatty alcohol according to the present application has a significant fungistatic and fungicidal effect on fungi, thereby reducing the occurrence of eggplant gray mold and eggplant blight in the treatment group, and significantly increasing the yield of eggplants.

[0316] Example 14: Effect of the preparation containing a higher fatty alcohol on the prevention and treatment of diseases in cowpea fields

[0317] Sampling: Sample S of the treatment group 1 is the water emulsion prepared in Example 1; sample C of the treatment group 2 is the water emulsion prepared from test agent 3 in Example 2;

[0318] Crop: cowpea big meat cowpea

[0319] Location: Sanxiang Town, Zhongshan City, Guangdong Province

[0320] Method: In the production base, 2 mu of the treatment group 1, 2 mu of the treatment group 2, and 2 mu of the control group are selected for production. The treatment group 1 and the treatment group 2 are produced as follows: the sample S is diluted 100 times with water, the cowpea seedlings are immersed in the solution, and then planted. The sample S is diluted with water and sprayed. The seedlings, flowering stage, fruiting stage, and harvesting stage are each sprayed once. The dilution multiples of the seedlings, flowering stage, fruiting stage, and harvesting stage are 800 times, 600 times, 500 times, and 500 times, respectively. The sample dosage is 60 mL per mu each time. At the same time of the treatment of the treatment group, the control group is sprayed with the same amount of water.

[0321] Comparison of use effect:

[0322] The cowpeas in the treatment group 1 and the treatment group 2 treated with the preparation containing a higher fatty alcohol according to the present application have good plant growth conditions, lush leaves, and a significant advantage in the number of flowers and pods, and more flowers and pods in the late harvesting stage.

[0323] Table 25: Test effect of cowpea treated with the preparation containing a higher fatty alcohol according to the present application

[0324] The above table shows that the cowpea treated with the preparation containing the higher fatty alcohol according to the application has good plant growth, more flowering and podding, and high yield, and the yield per mu of the treatment group 1 and the treatment group 2 is 3297 kg and 3184 kg respectively, which is increased by 17.8% and 13.8% respectively compared with the yield per mu of the control group 2798 kg; at the same time, the treatment group has less disease occurrence, while the control group is more affected by cowpea damping-off and cowpea rust in the whole growth process. The pathogen of cowpea rust is Phakopsora pachyrizi, which belongs to Basidiomycota fungi, and the pathogen of cowpea damping-off is Rhizoctonia solani, which belongs to Deuteromycota fungi, and cowpea is affected by these two diseases, resulting in reduced yield. It can be seen from the comparison that the preparation containing the higher fatty alcohol according to the application has obvious fungicidal effect on these two fungi, and after use, the treatment group reduces the occurrence of cowpea damping-off and cowpea rust, and significantly improves the yield of cowpea.

[0325] Example 15: Effect of the preparation containing the higher fatty alcohol on preventing and treating diseases in pepper field

[0326] Sampling: sample S is the aqueous emulsion prepared in example 1;

[0327] Crop: processed pepper Fuguihong;

[0328] Location: Karamay Agricultural Development Zone, Xinjiang;

[0329] Method: 40 mu of treatment group and 40 mu of control group are selected for production in the production base, and the treatment group is produced as follows: sample S is diluted 800 times with water, and then sprayed, and sprayed once every 15 days from the seedling stage, and the sample dosage is 60 mL per mu each time; at the same time of treating the treatment group, the control group is sprayed with the same amount of water.

[0330] Comparison of use effect:

[0331] The pepper in the treatment group treated with the preparation containing the higher fatty alcohol according to the application has no disease damage in the whole growth process, and the plant is healthy in appearance, the branches are strong and vigorous, the new and old leaves have good expansion, the leaves are tender green, and the peppers are full of the plant.

[0332] Table 26: Test effect of pepper using the preparation containing the higher fatty alcohol according to the application

[0333] The above table shows that the pepper treated by the preparation containing the higher fatty alcohol according to the present application has strong plant, more fruits, and high yield, which is 23.2% higher than that of the control group, and no obvious diseases and pests, while the control group is invaded by pepper blight during the whole growth process. The pathogen of pepper blight is pepper pythium, which is a pathogenic fungus belonging to the phylum of flagellum and the genus of pythium. The growth and yield of pepper are seriously affected by the invasion of pepper blight. It can be seen from the comparison that the preparation containing the higher fatty alcohol according to the present application has obvious fungistatic and fungicidal effects on this fungus, and the treatment group after use reduces the occurrence of pepper blight and significantly improves the yield of pepper.

[0334] Example 16: Effect of the preparation containing the higher fatty alcohol on the prevention and treatment of diseases in the apple field

[0335] Sampling: sample S is the aqueous emulsion prepared in example 1;

[0336] Crop: red apple;

[0337] Location: Karamay Agricultural Development Zone, Xinjiang;

[0338] Method: 20 mu of treatment group and 20 mu of control group are selected for production in the production base. The treatment group is produced as follows: after dilution with sample S, spray at 800 times in the young fruit stage and 600 times in the fruit expansion stage, spray once every 15 days, and the sample usage is 100 mL / mu each time; at the same time of treatment of the treatment group, the control group is sprayed with the same amount of water.

[0339] Comparison of use effect:

[0340] The apple treated by the preparation containing the higher fatty alcohol according to the present application has no disease damage during the whole growth process, more fruits on the plant, healthier and glossier appearance, better appearance, fewer rotten and deformed fruits, and earlier fruit color change of the plant in the treatment group.

[0341] Table 27: Test effect of apple using the preparation containing the higher fatty alcohol according to the present application

[0342] The above table shows that the apple treated by the preparation containing the higher fatty alcohol according to the present application has strong plant, more fruits, and high yield, which is 23.2% higher than that of the control group, and no obvious diseases and pests, while the control group is invaded by pepper blight during the whole growth process. The pathogen of pepper blight is pepper pythium, which is a pathogenic fungus belonging to the phylum of flagellum and the genus of pythium. The growth and yield of pepper are seriously affected by the invasion of pepper blight. It can be seen from the comparison that the preparation containing the higher fatty alcohol according to the present application has obvious fungistatic and fungicidal effects on this fungus, and the treatment group after use reduces the occurrence of pepper blight and significantly improves the yield of pepper.

[0343] Example 17: Disease control effect of the preparation containing higher fatty alcohol on litchi trees

[0344] Sampling: sample S of treatment group 1 was the water emulsion prepared in Example 1; sample C of treatment group 2 was the water emulsion prepared from agent 3 in Example 2;

[0345] Crop: litchi with a Guime taste;

[0346] Location: DaJing town, GaoZhou city, Guangdong province;

[0347] Method: 10 mu of treatment group 1, 10 mu of treatment group 2, and 10 mu of the control group were selected for production in the production base. The treatment group 1 and the treatment group 2 were produced as follows: from the beginning of the spring fruit tree sprouting tender leaves, the sample was diluted 1000 times with water and sprayed, and the spraying was performed once every 15 days, with a sample dosage of 100 mL per mu; at the same time of the treatment of the treatment group, the control group was sprayed with the same amount of water.

[0348] Comparison of use effects:

[0349] The fruit trees treated with the water emulsion of the preparation containing higher fatty alcohol of the application in the treatment group had strong tree vigor, dark green and lustrous leaf color, no obvious disease and pest damage, and less split and rotten fruits. However, the fruit trees in the control group had litchi anthracnose and litchi downy mildew, some of the fruit trees had weak tree vigor, and had more defoliation, fruit drop, split and rotten fruits.

[0350] Table 28: Test effect of litchi using the preparation containing higher fatty alcohol of the application

[0351] The above table shows that the fruit trees treated with the preparation containing higher fatty alcohol of the application had vigorous growth and less split and rotten fruits. The litchi yield per mu of the treatment group 1 and the treatment group 2 was 314 kg and 326 kg, respectively, which was 28.2% and 33.1% higher than that of the control group (245 kg). Both the treatment group 1 and the treatment group 2 had no obvious disease and pest damage, while the control group had litchi anthracnose and litchi downy mildew. Litchi anthracnose was caused by infection of Colletotrichum gloeosporioides or Glomerella cingulata, and these fungi were the direct cause of the disease. The pathogen of litchi downy mildew was Peronophythora litchii, which was an oomycete that could affect the tender leaves, tender branches, flower spikes and fruits of litchi, causing significant yield loss. The preparation containing higher fatty alcohol (n-decanol or n-dodecanol) of the application had obvious bacteriostatic and bactericidal effects on both the fungi causing litchi anthracnose and the oomycete causing litchi downy mildew, thereby reducing the occurrence of litchi anthracnose and litchi downy mildew in the treatment group and reducing the possible yield loss.

[0352] The results of the test researches on the growth, physiological and biochemical effects and transcription sequencing analysis of the model organism Saccharomyces cerevisiae after high-grade fatty alcohol stress in Example 4 can be seen that the high-grade fatty alcohol destroys the cell wall integrity of Saccharomyces cerevisiae cells, causes the cell structure to be damaged, the endogenous substances to flow out, and the function of mitochondria of Saccharomyces cerevisiae cells to be destroyed, thereby causing the cell damage and death, and showing that the high-grade fatty alcohol has the inhibition and killing effect on eukaryotic cells.

[0353] As can be seen from Example 5 and Example 6, it is determined in the laboratory tests that the preparation containing high-grade fatty alcohol of the present application has certain inhibition and killing effect on the growth of plant pathogenic fungi such as Fusarium graminearum, Fusarium oxysporum and Colletotrichum gloeosporioides, which is consistent with the results of the growth of the model organism Saccharomyces cerevisiae affected by the high-grade fatty alcohol in Example 4, and it is shown that the high-grade fatty alcohol can directly destroy the cell wall and mitochondria of fungal cells, cause the cell structure to be damaged, the endogenous substances to flow out and the dysfunction of mitochondria, cause the growth inhibition, damage and death of fungal cells, and show that the high-grade fatty alcohol has the bacteriostatic and bactericidal effect on plant pathogenic fungi.

[0354] As can be seen from Example 7, it is determined in the laboratory tests that the preparation containing high-grade fatty alcohol of the present application has certain inhibition and killing effect on the growth of the plant pathogenic oomycete Peronophythora litchii.

[0355] As can be seen from Example 8 to Example 17, the emulsifier containing high-grade fatty alcohol of the present application can be used on a variety of crops such as wheat, peanut, citrus, sugarcane, cucumber, eggplant, cowpea, pepper, apple and litchi, and can effectively prevent and control a variety of plant diseases caused by plant pathogenic fungi and oomycetes, and improve the yield, which shows that the emulsifier containing high-grade fatty alcohol of the present application can inhibit and kill fungi and oomycetes, and effectively prevent and control the related diseases caused by plant pathogenic fungi and oomycetes.

[0356] In summary of Example 4 to Example 17, the preparation containing high-grade fatty alcohol of the present application can directly destroy the cell wall and mitochondria of fungal cells, cause the cell structure to be damaged, the endogenous substances to flow out and the dysfunction of mitochondria, cause the growth inhibition, damage and death of fungal cells, and also can inhibit the growth of oomycetes, and show that the preparation containing high-grade fatty alcohol has the bacteriostatic and bactericidal effect on fungi and oomycetes. The preparation containing high-grade fatty alcohol of the present application can be used on plants to inhibit and kill plant pathogenic fungi and oomycetes, thereby effectively preventing and controlling the diseases caused by plant pathogenic fungi and oomycetes.

[0357] The above embodiments are only preferred embodiments of the present application, and cannot be used to limit the protection scope of the present application, and any non-essential changes and substitutions made by those skilled in the art on the basis of the present application shall fall within the protection scope of the present application.

Claims

1. Use of higher aliphatic alcohols for the preparation of preparations for the inhibition and destruction of fungi, characterized in that, The high-grade fatty alcohol includes one of n-decanol and n-dodecanol or a mixture of both.

2. Use according to claim 1, wherein The preparation realizes the application in the bacteriostatic and fungicidal aspects of fungi by destroying the cell wall integrity, causing the cell structure to be damaged, the endogenous substances to outflow, causing the growth inhibition, damage and death of the fungal cells.

3. The use according to claim 1, wherein The preparation realizes the application in the bacteriostatic and fungicidal aspects of fungi by destroying the cell wall integrity, causing the cell structure to be damaged, the endogenous substances to outflow, causing the growth inhibition, damage and death of the fungal cells.

4. Use according to any one of claims 1 to 3, wherein The preparation includes one of water emulsion, emulsifiable concentrate, suspension, wettable powder, powder, granule, tablet, water agent, sprayable solution, concentrated emulsion, aerosol and seed coating agent.

5. Use of higher aliphatic alcohols for the preparation of a preparation for the control of diseases caused by phytopathogenic fungi, characterized in that, The high-grade fatty alcohol includes one of n-decanol and n-dodecanol or a mixture of both.

6. Use according to claim 5, wherein The preparation realizes the prevention and control of the diseases caused by the phytopathogenic fungi by destroying the cell wall integrity of the phytopathogenic fungi, causing the cell structure to be damaged, the endogenous substances to outflow, causing the growth inhibition, damage and death of the fungal cells.

7. The use according to claim 5, wherein the compound is ###0002### The preparation realizes the prevention and control of the diseases caused by the phytopathogenic fungi by destroying the cell wall integrity of the phytopathogenic fungi, causing the cell structure to be damaged, the endogenous substances to outflow, causing the growth inhibition, damage and death of the fungal cells.

8. Use according to any one of claims 5 to 7, wherein the compound is of formula (I) ###0001### (I) or a pharmaceutically acceptable salt thereof. The preparation includes one of water emulsion, emulsifiable concentrate, suspension, wettable powder, powder, granule, tablet, water agent, sprayable solution, concentrated emulsion, aerosol and seed coating agent.

9. Use of higher aliphatic alcohols for the preparation of a preparation for the inhibition and killing of Oomycetes, characterized in that, The high-grade fatty alcohol includes one of n-decanol and n-dodecanol or a mixture of both.

10. Use of higher aliphatic alcohols for the preparation of a formulation for the control of diseases caused by phytopathogenic Oomycetes, characterized in that, The high-grade fatty alcohol includes one of n-decanol and n-dodecanol or a mixture of both.

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