Antifungal effect of antifungal emulsion and its application in prevention of chestnut decay

Abstract

The application discloses an antifungal effect of an antifungal emulsion and application of the antifungal emulsion in prevention and treatment of chestnut corruption, and belongs to the technical field of food preservation. The application is used for preventing and treating corruption caused by fungal infection during the collection and storage period of chestnuts. The antifungal emulsion comprises an antifungal component, an emulsifier and distilled water. The antifungal component comprises at least one of plant essential oil and active components of plant essential oil. The plant essential oil is at least one of thyme essential oil, clove essential oil and perilla essential oil. The active components of plant essential oil are at least one of thymol, eugenol and perillaldehyde. The application has good prevention and treatment effect on fungal infection of chestnuts during the collection and storage period, and has the advantages of being green, safe and efficient.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of food preservation technology, in particular to the antifungal effect of antifungal emulsion and its application in the prevention of chestnut corruption. BACKGROUND

[0002] Chestnut, commonly known as chestnut, alias Mao Li, chestnut, etc., belongs to nuts, is a food with very high nutritional value, and is one of important economic crops. The fresh chestnut has a high water content, and is easily infected by fungi during storage, leading to corruption and affecting the yield and quality of chestnut. After the spiny bracts are harvested, the chestnut fruits are usually stripped and harvested after maturing by stacking. This harvesting mode can further aggravate the infection and spread of the corruption fungi, causing the chestnut to rot during the stacking process, and the corruption will further develop during the subsequent storage process.

[0003] The preservation period of chestnut includes the collection and storage period and the storage period. The collection and storage period is composed of the storage and maturation of spiny bracts. The storage period usually uses preservation technology to store mature chestnut. The preservation methods include but are not limited to traditional sand storage, low-temperature storage, irradiation treatment, chemical preservative, high-molecular coating, and modified atmosphere packaging. The above preservation methods have disadvantages such as high cost, harmful substance residue, and influence on food flavor. Therefore, preventing and treating the infection of corruption fungi in the collection and storage period of chestnut is an important research direction. Plant essential oil is an oil-like liquid with volatility and special aroma extracted from flowers, buds, leaves, roots, seeds, branches and fruits of plants, which has the advantages of green, safe and efficient. Plant essential oil and its active ingredients are widely used in the prevention of food corruption fungi. The current application of preservative ingredients is mainly concentrated in the storage period of chestnut, and there is no concern about the fungal infection of chestnut in the collection and storage period. SUMMARY

[0004] In view of the above defects of the prior art, in the first aspect of the present application, the application of antifungal emulsion in preventing and treating chestnut corruption is provided. The chestnut corruption is caused by fungal infection leading to corruption of chestnut kernel or spiny bract. The antifungal emulsion includes antifungal ingredients, emulsifiers and distilled water. The antifungal ingredients include at least one of plant essential oil and plant essential oil active ingredients. The plant essential oil is at least one of thyme essential oil, clove essential oil and perilla essential oil. The plant essential oil active ingredient is at least one of thymol, eugenol and perillaldehyde. The application time of the antifungal emulsion for preventing and treating chestnut corruption is the collection and storage period of chestnut.

[0005] Preferably, the antifungal emulsion includes 0.05% to 7.5% of antifungal ingredients, 0.05% to 5% of emulsifiers and the rest of distilled water by mass percentage.

[0006] Preferably, the emulsifier is at least one of Tween 20, Tween 80, Span 80, sucrose fatty acid ester, monoglyceride fatty acid glyceride.

[0007] Preferably, the fungus infected fungus includes at least one of Neofusicoccum parvum, Botryosphaeria dothidea, Fusarium proliferatum, Colletotrichum siamense, Acrospeira mirabilis, Alternaria alternata, Aspergillus tubingensis, Lasiodiplodia pseudotheobromae.

[0008] Further preferably, when the fungus infected fungus is Neofusicoccum parvum, the minimum inhibitory concentration of thyme essential oil on Neofusicoccum parvum is 0.50 μL / mL, and the minimum bactericidal concentration is 0.75 μL / mL; The minimum inhibitory concentration of thymol on Neofusicoccum parvum is 0.25 mg / mL, and the minimum bactericidal concentration is 0.50 mg / mL.

[0009] Further preferably, when the fungus infected fungus is Botryosphaeria dothidea, the minimum inhibitory concentration of thyme essential oil on Botryosphaeria dothidea is 0.50 μL / mL, and the minimum bactericidal concentration is 0.75 μL / mL; The minimum inhibitory concentration of thymol on Botryosphaeria dothidea is 0.50 mg / mL, and the minimum bactericidal concentration is 0.75 mg / mL.

[0010] Further preferably, when the fungus infected fungus is Fusarium proliferatum, the minimum inhibitory concentration of thyme essential oil on Fusarium proliferatum is 0.50 μL / mL, and the minimum bactericidal concentration is 0.75 μL / mL; The minimum inhibitory concentration of thymol on Fusarium proliferatum is 0.50 mg / mL, and the minimum bactericidal concentration is 0.75 mg / mL.

[0011] Further preferably, when the fungus infected fungus is Neofusicoccum parvum, the minimum inhibitory concentration of clove essential oil on Neofusicoccum parvum is 1.00 μL / mL, and the minimum bactericidal concentration is 1.50 μL / mL; The minimum inhibitory concentration of eugenol on Neofusicoccum parvum is 0.75 μL / mL, and the minimum bactericidal concentration is 1.00 μL / mL.

[0012] Further preferably, when the fungus infected by the fungus is Botryosphaeria dothidea, the minimum inhibitory concentration of clove essential oil on Botryosphaeria dothidea is 1.00 μL / mL, and the minimum fungicidal concentration is 1.50 μL / mL; the minimum inhibitory concentration of eugenol on Botryosphaeria dothidea is 0.75 μL / mL, and the minimum fungicidal concentration is 1.00 μL / mL.

[0013] Further preferably, when the fungus infected by the fungus is Botryosphaeria dothidea, the minimum inhibitory concentration of clove essential oil on Botryosphaeria dothidea is 1.00 μL / mL, and the minimum fungicidal concentration is 1.50 μL / mL; the minimum inhibitory concentration of eugenol on Botryosphaeria dothidea is 0.75 μL / mL, and the minimum fungicidal concentration is 1.00 μL / mL.

[0014] Further preferably, when the fungus infected by the fungus is Botryosphaeria dothidea, the minimum inhibitory concentration of clove essential oil on Botryosphaeria dothidea is 1.00 μL / mL, and the minimum fungicidal concentration is 1.50 μL / mL; the minimum inhibitory concentration of eugenol on Botryosphaeria dothidea is 0.75 μL / mL, and the minimum fungicidal concentration is 1.00 μL / mL.

[0015] Further preferably, when the fungus infected by the fungus is Botryosphaeria dothidea, the minimum inhibitory concentration of clove essential oil on Botryosphaeria dothidea is 1.00 μL / mL, and the minimum fungicidal concentration is 1.50 μL / mL; the minimum inhibitory concentration of eugenol on Botryosphaeria dothidea is 0.75 μL / mL, and the minimum fungicidal concentration is 1.00 μL / mL.

[0016] Further preferably, when the fungus infected by the fungus is Botryosphaeria dothidea, the minimum inhibitory concentration of clove essential oil on Botryosphaeria dothidea is 1.00 μL / mL, and the minimum fungicidal concentration is 1.50 μL / mL; the minimum inhibitory concentration of eugenol on Botryosphaeria dothidea is 0.75 μL / mL, and the minimum fungicidal concentration is 1.00 μL / mL.

[0017] Preferably, the specific application method of the anti-fungal emulsion in preventing and treating chestnut corruption includes the following steps: the anti-fungal emulsion is diluted to obtain a preservation liquid, the preservation liquid is applied to the surface of the chestnut in the storage period to form a drug film containing the anti-fungal component on the surface, and the drug film is used for preventing and treating the corruption of the chestnut caused by the fungal infection in the storage period.

[0018] Further preferably, in the preservation liquid, the concentration of the anti-fungal component is greater than or equal to the minimum inhibitory concentration of the fungus infection.

[0019] More preferably, the amount of the preservative solution applied is 2% to 5% of the fresh weight of the chestnuts.

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

[0021] This invention provides the application of an antifungal emulsion in the prevention and control of chestnut spoilage. This application involves intervention during the chestnut harvest and storage period, demonstrating excellent control over fungal infection causing spoilage of chestnut kernels or burrs. The antifungal components in the emulsion are extracted from natural plants. Compared to chemical preservation methods, this invention avoids the harm to human health and environmental damage caused by chemical residues, offering advantages of safety, effectiveness, and environmental friendliness. Attached Figure Description

[0022] Figure 1 The results of the effects of different concentrations of thyme essential oil on the mycelial growth of Clostridium difficile;

[0023] Figure 2 The results show the effect of different concentrations of thymol on the mycelial growth of Clostridium difficile;

[0024] Figure 3 Figure A shows the results of chestnut kernels being infected by Clostridium difficile under different treatment conditions; Figure B shows the statistical results of the rot rate of each group.

[0025] Figure 4 Figure A shows the statistical results of chestnut kernel rot rate when quantity is the benchmark; Figure B shows the statistical results of chestnut kernel rot rate when quality is the benchmark.

[0026] Figure 5 The results show the effects of different concentrations of thyme essential oil on the mycelial growth of *Vitis vinifera*.

[0027] Figure 6 The results show the effect of different concentrations of thymol on the mycelial growth of *Botrytis cinerea*.

[0028] Figure 7 The results show the effect of different concentrations of thyme essential oil on the growth of Fusarium effusum mycelium;

[0029] Figure 8 The results show the effect of different concentrations of thymol on the growth of Fusarium effusum mycelium.

[0030] In the above figure, CK represents the blank group, Model represents the model group, Thyme represents thyme essential oil, and Thymol represents thymol. Detailed Implementation

[0031] The application will be further described in the following by way of examples without thereby limiting the application to the examples described. The experimental methods in the following examples, for which no specific conditions are indicated, are carried out according to routine methods and conditions, or according to the instructions of the commercial suppliers.

[0032] Example 1

[0033] Study on the in vitro antifungal effect of thyme essential oil and thymol on Chinese chestnut against Neofusicoccum parvum

[0034] Neofusicoccum parvum isolated from rotten Chinese chestnut was inoculated on PDA medium and cultured at 28℃ for 3 days. (1) Thyme essential oil emulsion was prepared by dissolving 7.5% thyme essential oil and 0.1% Tween 20 in the rest of distilled water. The thyme essential oil emulsion was diluted with PDA medium to prepare plates containing 0.10, 0.25, 0.50, 0.75 μL / mL of thyme essential oil as experimental groups (Thyme), and plates without thyme essential oil as blank group (CK). (2) Thymol emulsion was prepared by dissolving 5% thymol and 0.1% Tween 20 in the rest of distilled water. The thymol emulsion was diluted with PDA medium to prepare plates containing 0.05, 0.10, 0.25, 0.50 mg / mL of thymol as experimental groups (Thymol), and plates without thymol as blank group (CK). The sterile puncher was used to punch the colony at the edge of the growing colony, and the mycelium was inoculated in the center of the plate containing the drug and placed in a constant temperature incubator at 28℃ for inverted culture. Three replicates were set for each treatment group, and the mycelial growth diameter was measured every 24 h using the cross method. The growth curve of Neofusicoccum parvum was drawn, and the minimum inhibitory concentration (MIC) and the minimum fungicidal concentration (MFC) of thyme essential oil and thymol were obtained.

[0035] As shown in Figure 1 , thyme essential oil has good antibacterial effect in vitro on Chinese chestnut. Thyme essential oil with a concentration of 0.50 μL / mL can inhibit the growth of Neofusicoccum parvum, and thyme essential oil with a concentration of 0.75 μL / mL can completely kill the mycelium of Neofusicoccum parvum. The minimum inhibitory concentration (MIC) of thyme essential oil on Neofusicoccum parvum is 0.50 μL / mL, and the minimum fungicidal concentration (MFC) is 0.75 μL / mL.

[0036] As shown in Figure 2As shown, thymol exhibits excellent antibacterial effects in chestnuts even in vitro. A concentration of 0.25 mg / mL of thymol can inhibit the growth of *Neofusicoccum parvum*, and a concentration of 0.50 mg / mL of thymol can completely kill *Neofusicoccum parvum* hyphae. The minimum inhibitory concentration (MIC) of thymol against *Neofusicoccum parvum* is 0.25 mg / mL, and the minimum bactericidal concentration (MFC) is 0.50 mg / mL.

[0037] Example 2

[0038] In vivo antifungal effects of antifungal emulsions containing thyme essential oil and thymol on chestnut kernels:

[0039] The cultured mycelia of *Neofusicoccum parvum* were homogenized with sterile water to prepare a mycelial suspension of 1 mg / mL. Chestnut kernels with the peel removed were washed with sterile water and cut into uniform small pieces weighing 1 g each. (1) By mass percentage, 7.5% thyme essential oil and 0.1% Tween 20 were dissolved in the remaining distilled water to prepare a thyme essential oil emulsion. The thyme essential oil emulsion was diluted with distilled water to a final concentration of 0.50, 0.75, and 3.75 μL / mL (i.e., MIC, MFC, and 5 times MFC) to obtain thyme essential oil solutions. The treatment groups were given the mycelial suspension and thyme essential oil solutions at 5% of the chestnut kernel weight (referred to as 0.5-3%, 0.75-3%, and 3.75-3% respectively, based on the difference in thyme essential oil concentration). (2) By mass percentage, 5% thymol and 0.1% Tween 20 were dissolved in the remaining distilled water to prepare a thymol emulsion. The thymol emulsion was diluted with distilled water to a final thymol concentration of 0.25, 0.50, and 2.50 mg / mL (i.e., MIC, MFC, and 5 times MFC) to obtain thymol solutions. The treatment groups were given 5% of the weight of chestnut kernels, respectively, of the mycelial suspension and the thymol solution (referred to as 0.25-3, 0.5-3, and 2.5-3 according to the difference in thymol concentration). The model group (M3) was given only mycelial suspension, and the blank group (CK) was not treated. Each treatment group had 3 replicates, and 5 chestnut pieces were placed on each replicate plate. The plates were placed at room temperature until all the model groups were infected, and the decay rate of each group was counted.

[0040] like Figure 3 As shown, all model groups were infected by *Clostridium difficile* under experimental conditions. MFC and thyme essential oil with 5 times the amount of MFC significantly inhibited *Clostridium difficile* infection in chestnuts, with 5 times the amount of MFC showing better antibacterial effect. MFC and thymol with 5 times the amount of MFC also significantly inhibited *Clostridium difficile* infection in chestnuts, with 5 times the amount of MFC showing better antibacterial effect.

[0041] Example 3

[0042] Study on the Antifungal Emulsion Containing Thyme Oil and Thymol for Preserving Chinese Chestnut during Harvesting and Storage Period

[0043] Take the cultured Neofusicoccum parvum mycelium and homogenize it into a 1 mg / mL bacterial suspension with sterile water. (1) Dissolve 7.5% thyme oil, 2.5% Tween 80, and 2.5% Span 80 in the remaining distilled water to prepare a thyme oil emulsion. Dilute the thyme oil emulsion with distilled water to a final concentration of 0.75 μL / mL and 3.75 μL / mL of thyme oil. Divide the newly harvested Chinese chestnut burrs into 30 groups each for the blank group (CK), the model group (Model), the low-dose thyme oil group (Thyme-L, 0.75 μL / mL), and the high-dose thyme oil group (Thyme-H, 3.75 μL / mL). Each treatment group has three replicates. Spray the bacterial suspension and thyme oil solution on each group according to 2% of the weight of the Chinese chestnut. The model group is only sprayed with the bacterial suspension. The blank group is not treated. (2) Dissolve 5% thymol, 2.5% Tween 80, and 2.5% Span 80 in the remaining distilled water to prepare a thymol emulsion. Dilute the thymol emulsion with distilled water to a final concentration of 0.50 mg / mL and 2.50 mg / mL of thymol. Divide the newly harvested Chinese chestnut burrs into 30 groups each for the blank group (CK), the model group (Model), the low-dose thymol group (Thymol-L, 0.50 mg / mL), and the high-dose thymol group (Thymol-H, 2.50 mg / mL). Each treatment group has three replicates. Spray the bacterial suspension and thymol solution on each group according to 2% of the weight of the Chinese chestnut. The model group is only sprayed with the bacterial suspension. The blank group is not treated. Observe the Chinese chestnut rotting every 4 days, and calculate the Chinese chestnut kernel rotting rate based on two methods; Method A is based on the number, and the Chinese chestnut kernel rotting rate is the ratio of the number of rotten Chinese chestnut kernels to the total number of Chinese chestnut kernels. Method B is based on the mass, and the Chinese chestnut kernel rotting rate is the ratio of the mass of rotten Chinese chestnut kernels to the total mass of Chinese chestnut kernels.

[0044] As shown in Figure 4 , the high-dose thyme oil group has the best preservation effect after 6 observation periods, and the Chinese chestnut rotting rate is still less than 30% under the two statistical methods, which has a good effect on preventing fungal infection of Chinese chestnut fruit during the harvesting and storage period.

[0045] As shown in Figure 4As shown, the high-dose thymol group showed the best antiseptic effect after 6 observation cycles. Under both statistical methods, the chestnut rot rate was still less than 30% at this time, which has a good antiseptic effect on fungal infection of chestnuts caused during the harvest and storage period.

[0046] Example 4

[0047] In vitro antifungal activity of antifungal emulsions containing thyme essential oil and thymol against chestnut:

[0048] Botryosphaeria dothidea, isolated from rotten chestnuts, was inoculated onto PDA medium and cultured at 28°C for 6 days. (1) 7.5% thyme essential oil and 0.1% Tween 20 were dissolved in the remaining distilled water by weight percentage to prepare thyme essential oil emulsion. The thyme essential oil emulsion was diluted with PDA medium to prepare plates with a final concentration of 0.10, 0.25, 0.50, and 0.75 μL / mL of thyme essential oil and used as experimental groups (Thyme), while plates without thyme essential oil were used as blank groups (CK). (2) 5% thymol and 0.1% Tween 20 were dissolved in the remaining distilled water by weight percentage to prepare thymol emulsion. Thymol emulsion was diluted with PDA medium to prepare thymol-containing plates with final concentrations of 0.10, 0.25, 0.50, and 0.75 mg / mL, serving as the experimental group (Thymol). Plates without thymol served as the control group (CK). Using a sterile punch, holes were made at the edges of vigorous colonies, and mycelial discs were inoculated into the center of the treated plates. The plates were then incubated upside down at 28°C. Each treatment group was replicated in triplicate, and mycelial diameter was measured every 24 hours using the cross-sectional method. Growth curves of *Botryosphaeria dothidea* were plotted to determine the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of thyme oil and thymol.

[0049] like Figure 5 As shown, thyme essential oil exhibits excellent antibacterial effects in chestnuts even without in vitro exposure. A concentration of 0.50 μL / mL of thyme essential oil can inhibit the growth of *Botryosphaeria dothidea*, and a concentration of 0.75 μL / mL of thyme essential oil can completely kill *Botryosphaeria dothidea* hyphae. The minimum inhibitory concentration (MIC) of thyme essential oil against *Botryosphaeria dothidea* is 0.50 μL / mL, and the minimum bactericidal concentration (MFC) is 0.75 μL / mL.

[0050] like Figure 6As shown, thymol exhibits excellent antibacterial effects in chestnuts even in vitro. A concentration of 0.50 mg / mL of thymol can inhibit the growth of *Botryosphaeria dothidea*, and a concentration of 0.75 mg / mL of thymol can completely kill *Botryosphaeria dothidea* hyphae. The minimum inhibitory concentration (MIC) of thymol against *Botryosphaeria dothidea* is 0.50 mg / mL, and the minimum bactericidal concentration (MFC) is 0.75 mg / mL.

[0051] Example 5

[0052] In vitro antifungal activity of antifungal emulsions containing thyme essential oil and thymol against chestnut:

[0053] Fusarium proliferatum, isolated from rotten chestnuts, was inoculated onto PDA medium and cultured at 28°C for 6 days. (1) Thyme essential oil emulsion was prepared by dissolving 7.5% thyme essential oil and 0.1% Tween 20 in the remaining distilled water by weight percentage. The thyme essential oil emulsion was diluted with PDA medium to prepare plates with a final concentration of 0.10, 0.25, 0.50, and 0.75 μL / mL of thyme essential oil and used as experimental groups (Thyme), while plates without thyme essential oil were used as blank groups (CK). (2) Thymol emulsion was prepared by dissolving 5% thymol and 0.1% Tween 20 in the remaining distilled water by weight percentage. Thymol emulsion was diluted with PDA medium to prepare thymol-containing plates with final concentrations of 0.10, 0.25, 0.50, and 0.75 mg / mL, serving as the experimental group (Thymol). Plates without thymol served as the control group (CK). Using a sterile punch, holes were made at the edges of vigorous colonies, and mycelial discs were inoculated into the center of the treated plates. The plates were then incubated upside down at 28°C. Each treatment group was replicated in triplicate, and mycelial diameter was measured every 24 hours using the cross-hatching method. Growth curves of *Fusarium proliferatum* were plotted to determine the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of thyme oil and thymol.

[0054] like Figure 7 As shown, thyme essential oil exhibits excellent antibacterial effects in chestnuts even without in vitro exposure. A concentration of 0.50 μL / mL of thyme essential oil can inhibit the growth of *Fusarium proliferatum*, and a concentration of 0.75 μL / mL of thyme essential oil can completely kill *Fusarium proliferatum* hyphae. The minimum inhibitory concentration (MIC) of thyme essential oil against *Fusarium proliferatum* is 0.50 μL / mL, and the minimum bactericidal concentration (MFC) is 0.75 μL / mL.

[0055] likeFigure 8 As shown, thymol exhibits excellent antibacterial effects in chestnuts even in vitro. A concentration of 0.50 mg / mL of thymol can inhibit the growth of *Fusarium proliferatum*, and a concentration of 0.75 mg / mL of thymol can completely kill *Fusarium proliferatum* mycelia. The minimum inhibitory concentration (MIC) of thymol against *Fusarium proliferatum* is 0.50 mg / mL, and the minimum bactericidal concentration (MFC) is 0.75 mg / mL.

[0056] Example 6

[0057] The in vitro antifungal effects of clove oil, eugenol, perilla oil, and perillaldehyde on chestnut were tested using the same methods as in Examples 1, 4, and 5. Based on the results of Examples 1, 4, and 5, the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) of the antifungal components in the antifungal emulsion of this invention against *Neofusicoccum parvum*, *Botryosphaeria dothidea*, and *Fusarium proliferatum* are shown in Table 1.

[0058] Table 1: Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MFC) of different antifungal components against three major contaminating bacteria of chestnuts.

[0059]

[0060]

[0061] The results in Table 1 show that clove oil, eugenol, perilla oil, and perillaldehyde also exhibit good antibacterial activity against the three main fungi that cause chestnut spoilage. These can be applied to prevent spoilage during the chestnut harvesting and storage period.

[0062] Example 7

[0063] The specific methods for using antifungal emulsions to prevent chestnut spoilage are as follows:

[0064] The antifungal emulsion, by weight percentage, consists of 0.05% clove oil, 0.05% Tween 80, and the balance distilled water. After dilution, the antifungal emulsion yields a preservative solution with a clove oil concentration of 1.50 μL / mL. The preservative solution is applied to the surface of chestnuts during the harvest and storage period, forming a film containing antifungal components on the surface to prevent spoilage caused by fungal infection during the harvest and storage period.

[0065] Example 8

[0066] The specific methods for using antifungal emulsions to prevent chestnut spoilage are as follows:

[0067] The antifungal emulsion is composed of 7.5% eugenol, 5% Span 80 and the rest of distilled water in percentage of mass; the antifungal emulsion is diluted to obtain the preservative liquid with eugenol concentration of 1.50 μL / mL, the preservative liquid is applied to the surface of the chestnut in the storage period to form a drug film containing antifungal components on the surface, which is used for preventing and treating the decay of the chestnut caused by fungal infection in the storage period.

[0068] Example 9

[0069] The application of the antifungal emulsion in preventing and treating the decay of the chestnut is as follows:

[0070] The antifungal emulsion is composed of 7.5% perilla oil, 5% sucrose fatty acid ester and the rest of distilled water in percentage of mass; the antifungal emulsion is diluted to obtain the preservative liquid with perilla oil concentration of 1.50 μL / mL, the preservative liquid is applied to the surface of the chestnut in the storage period to form a drug film containing antifungal components on the surface, which is used for preventing and treating the decay of the chestnut caused by fungal infection in the storage period.

[0071] Example 10

[0072] The antifungal emulsion is composed of 7.5% perilla aldehyde, 5% monoglyceride fatty acid glycerol ester and the rest of distilled water in percentage of mass; the antifungal emulsion is diluted to obtain the preservative liquid with perilla aldehyde concentration of 0.75 μL / mL, the preservative liquid is applied to the surface of the chestnut in the storage period to form a drug film containing antifungal components on the surface, which is used for preventing and treating the decay of the chestnut caused by fungal infection in the storage period. The preferred embodiments of the present application are described in detail above. It should be understood that those skilled in the art can make many modifications and variations without creative work based on the concept of the present application. Therefore, any technical solution obtained by logical analysis, reasoning or limited experiment based on the prior art according to the concept of the present application should be within the protection scope defined by the claims.

Claims

1. The application of antifungal emulsion in preventing chestnut spoilage, characterized by: The chestnut spoilage is caused by fungal infection leading to the decay of chestnut kernels or burrs; the antifungal emulsion is applied during the chestnut harvest and storage period to prevent chestnut spoilage; the antifungal emulsion comprises an antifungal component, an emulsifier, and distilled water; the antifungal component comprises at least one of plant essential oils and active ingredients of plant essential oils; the plant essential oil is at least one of thyme essential oil, clove essential oil, and perilla essential oil; the active ingredient of the plant essential oil is at least one of thymol, eugenol, and perillaldehyde. The fungi that cause infection include at least one of Neofusicoccum parvum, Botryosphaeria dothidea, and Fusarium proliferatum. When the fungus infecting the fungus is *Clostridium neoformans*, the minimum inhibitory concentration (MIC) of thyme essential oil against *Clostridium neoformans* is 0.50 μL / mL, and the minimum bactericidal concentration (MBC) is 0.75 μL / mL; the MIC of thymol against *Clostridium neoformans* is 0.25 mg / mL, and the MBC is 0.50 mg / mL. When the fungus infecting the fungus is *Staphylococcus aureus*, the MIC of thyme essential oil against *Staphylococcus aureus* is 0.50 μL / mL, and the minimum bactericidal concentration (MBC) is 0.75 μL / mL. The minimum inhibitory concentration (MIC) of thymol against Staphylococcus aureus is 0.75 μL / mL, and the minimum bactericidal concentration (MBC) is 0.50 mg / mL; when the fungus infecting the fungus is Fusarium moniliforme, the MIC of thyme essential oil against Fusarium moniliforme is 0.50 μL / mL, and the minimum bactericidal concentration (MBC) of thymol against Fusarium moniliforme is 0.75 μL / mL. When the fungus infecting the fungus is *Clostridium microphyllum*, the minimum inhibitory concentration (MIC) of clove essential oil against *Clostridium microphyllum* is 1.00 μL / mL, and the minimum bactericidal concentration (MBC) is 1.50 μL / mL; the MIC of eugenol against *Clostridium microphyllum* is 0.75 μL / mL, and the MBC is 1.00 μL / mL. When the fungus infecting the fungus is *Staphylococcus aureus*, the MIC of clove essential oil against *Staphylococcus aureus* is 1.00 μL / mL, and the minimum bactericidal concentration (MBC) is 1.50 μL / mL. The minimum inhibitory concentration (MIC) of eugenol against Staphylococcus aureus is 0.75 μL / mL, and the minimum bactericidal concentration (MBC) is 1.00 μL / mL. When the fungus infecting the fungus is Fusarium moniliforme, the MIC of clove oil against Fusarium moniliforme is 0.75 μL / mL, and the MBC is 1.00 μL / mL. The MIC of eugenol against Staphylococcus aureus is 1.00 μL / mL, and the MBC is 1.50 μL / mL. When the fungus infecting the fungus is *Clostridium microphyllum*, the minimum inhibitory concentration (MIC) of perilla essential oil against *Clostridium microphyllum* is 1.00 μL / mL, and the minimum bactericidal concentration (MBC) is 1.5 μL / mL; the MIC of perilla aldehyde against *Clostridium microphyllum* is 0.50 μL / mL, and the MBC is 0.75 μL / mL. When the fungus infecting the fungus is *Botrytis cinerea*, the MIC of perilla essential oil against *Botrytis cinerea* is 0.25 μL / mL, and the MBC is 0.75 μL / mL. The minimum inhibitory concentration (MIC) of perilla aldehyde against Staphylococcus aureus is 0.25 μL / mL, and the minimum bactericidal concentration (MBC) is 0.50 μL / mL. When the fungus infecting the fungus is Fusarium moniliforme, the MIC of perilla essential oil against Fusarium moniliforme is 0.75 μL / mL, and the minimum bactericidal concentration (MBC) is 1.50 μL / mL. The MIC of perilla aldehyde against Fusarium moniliforme is 0.50 μL / mL, and the minimum bactericidal concentration (MBC) is 0.75 μL / mL. The concentration of the antifungal component is greater than or equal to the minimum inhibitory concentration against fungal infection.

2. The application according to claim 1, characterized in that: The antifungal emulsion comprises, by weight percentage, 0.05% to 7.5% antifungal ingredient, 0.05% to 5% emulsifier, and the balance being distilled water.

3. The application according to claim 1, characterized in that: The emulsifier is at least one of Tween 20, Tween 80, Span 80, sucrose fatty acid ester, and monoglyceride fatty acid glyceride.

4. The application according to any one of claims 1 to 3, characterized in that, The specific application method includes the following steps: the antifungal emulsion is diluted to obtain a preservative solution, which is then applied to the surface of chestnuts during the harvest and storage period to form a film containing antifungal components on the surface, used to prevent chestnuts from spoiling due to fungal infection during the harvest and storage period.

5. The application according to claim 4, characterized in that: The amount of the preservative solution applied is 2% to 5% of the fresh weight of the chestnuts.

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