Chrysanthemum bud-litsea cubeba compound essential oil as well as preparation method and application thereof

Abstract

The invention discloses chrysanthemum bud-litsea cubeba compound essential oil as well as a preparation method and application thereof, and belongs to the technical field of biology. The invention discloses chrysanthemum bud-litsea cubeba compound essential oil. The chrysanthemum bud-litsea cubeba compound essential oil is formed by compounding chrysanthemum bud essential oil and litsea cubeba essential oil according to the volume ratio of (1-3): (1-3). The chrysanthemum bud-litsea cubeba compound essential oil prepared by the invention is a green and natural bactericide, can be used as a safe substitute for synthesizing pesticides and is used for long-term prevention and treatment of stem rot of precious traditional Chinese medicinal materials due to the advantages that the chrysanthemum bud-litsea cubeba compound essential oil is non-toxic and volatile, and pathogenic bacteria are difficult to generate drug resistance and the like. According to the method, the raw materials of the chrysanthemum bud and the litsea cubeba are widely artificially planted, the essential oil content is high, the components are rich, the preparation process is simple, and a theoretical basis can be provided for high-value utilization of chrysanthemum bud and litsea cubeba resources while the method is applied to prevention and treatment of the anoectochilus formosanus stem rot.

Description

Technical Field

[0001] The invention relates to the field of biotechnology, and more particularly to a chrysanthemum-litsea cubeba composite essential oil and a preparation method and application thereof. Background Art

[0002] Anoectochilus roxburghii is a precious herbal medicine of the genus Anoectochilus in the Orchidaceae family. Due to its high medicinal value, it is known as the "king of medicine" and "black ginseng". However, due to excessive excavation, wild resources are on the verge of extinction, resulting in a very prominent contradiction between the supply and demand of Anoectochilus roxburghii. Artificial cultivation of Anoectochilus roxburghii is the only way to solve the current contradiction. It has been widely planted in Fujian, Zhejiang and other places in my country. However, Anoectochilus roxburghii is susceptible to diseases and pests during artificial cultivation, which affects its yield and quality; among them, stem rot of Anoectochilus roxburghii is a major disease in its artificial cultivation, and Fusarium oxysporum is the main pathogen. Fusarium oxysporum is a common plant pathogenic fungus and is listed as the third largest soil-borne pathogenic fungus in the world; it has a wide host range and can cause more than 100 plant wilt diseases such as melons, Solanaceae, bananas, cotton, legumes and flowers, and can cause root rot, stem rot and vascular wilt of many economic crops and Chinese medicinal materials. At present, the main method of controlling the stem rot of golden thread vine is chemical pesticides. Since the pathogen can cause diseases in different growth stages of golden thread vine, the number of times of pesticide spraying is relatively large, and the control effect is not ideal. At the same time, chemical pesticides have problems such as high toxicity, long-lasting residue, and easy drug resistance of pathogens. This will lead to safety hazards in the use of golden thread vine and restrict the sustainable and high-quality development of the golden thread vine industry. Compared with chemical pesticides, natural drug source pesticides have low toxicity, easy degradation, pollution-free, and not easy to develop drug resistance. Therefore, the development of natural drug substitutes for chemical pesticides is particularly important.

[0003] Essential oils are a type of secondary metabolites extracted from plants. They can be obtained from the flowers, leaves, stems, roots and other parts of plants through methods such as steam distillation. They are receiving more and more attention as safe alternatives to synthetic pesticides due to their antibacterial and bactericidal activities. Studies have confirmed that essential oils from various Lamiaceae plants, such as orchid, spearmint, peppermint, oregano, seagrass and rosemary, have inhibitory activity against Fusarium oxysporum. Although the above essential oils have been proven to be able to inhibit Fusarium oxysporum, due to their low essential oil content and differences in the strength of their antibacterial effects, there are still many problems in their use as natural antibacterial agents in the prevention and control of golden thread vine diseases.

[0004] Therefore, providing a chrysanthemum-litsea cubeba composite essential oil and a preparation method and application thereof is a problem that those skilled in the art urgently need to solve. Summary of the invention

[0005] In view of this, the present invention provides a composite essential oil of Chrysanthemum officinale and Litsea cubeba and a preparation method and application thereof, and uses the composite essential oil of Chrysanthemum officinale and Litsea cubeba as a safe substitute for synthetic pesticides; and solves the problems of high toxicity, long residual, and easy drug resistance of pathogens in the process of current chemical pesticides inhibiting Fusarium oxysporum, the main pathogen of golden thread rot disease.

[0006] Chrysanthemum rice is a perennial herbaceous plant of the genus Chrysanthemum in the Asteraceae family. It is made by frying the unopened buds of the fine variety of yellow chrysanthemum in Suichang, Lishui, Zhejiang. It is praised as one of the four major Huai medicines by the Chinese herbal medicine community and mainly contains a variety of flavonoid compounds and volatile oils (2%-5%). Chrysanthemum rice essential oil has good antibacterial, blood pressure lowering and blood lipid lowering effects. Litsea cubeba is a plant of the genus Litsea cubeba in the Lauraceae family. It is a traditional woody essential oil plant resource in my country, and its fruit contains 4%-7% volatile oil. Litsea cubeba essential oil has significant antibacterial, anti-inflammatory and insecticidal effects, and has wide application potential in the fields of food, medicine, medical beauty, chemical industry, etc. And the above two essential oils have significant characteristics that the raw materials are widely planted, the essential oil content is extremely high, the antibacterial activity is more significant, and the comprehensive cost is low. The present invention uses it as a safe substitute for synthetic pesticides in the prevention and control of golden thread lotus stem rot.

[0007] In order to achieve the above object, the present invention adopts the following technical solution:

[0008] A chrysanthemum bud-litsea cubeba composite essential oil is prepared by compounding chrysanthemum bud essential oil and litsea cubeba essential oil in a volume ratio of (1-3):(1-3).

[0009] Further, the preparation method of the chrysanthemum essential oil and the Litsea cubeba essential oil is as follows:

[0010] The dried chrysanthemum and litsea cubeba are respectively used as raw materials, and the raw materials are crushed and sieved through a 60-mesh sieve for later use; 50 g of the crushed and sieved raw materials are respectively weighed and placed in a round-bottom flask, and 10 to 40 times the volume of water is added, and after soaking for 0.5 to 1 hour, the flask is placed in a microwave extractor, and irradiated and extracted for 15 to 45 minutes under the condition of a microwave power of 320 to 640 W, and the extracted oil-water mixture is collected and dried and dehydrated to obtain chrysanthemum essential oil and litsea cubeba essential oil, respectively.

[0011] Further, a method for preparing the composite essential oil of Chrysanthemum officinale and Litsea cubeba, the specific steps are as follows:

[0012] (1) preparing chrysanthemum seed essential oil and litsea cubeba essential oil respectively;

[0013] (2) The prepared Chrysanthemum chrysanthemum essential oil and Litsea cubeba essential oil are mixed in a volume ratio of (1-3): (1-3) to obtain Chrysanthemum chrysanthemum-Litsea cubeba composite essential oil.

[0014] Furthermore, the application of the Chrysanthemum officinale-Litsea cubeba composite essential oil in preventing and treating roxburghii stem rot.

[0015] Furthermore, the Chrysanthemum officinale-Litsea cubeba composite essential oil is used in inhibiting Fusarium oxysporum.

[0016] It can be seen from the above technical scheme that, compared with the prior art, the present invention discloses a composite essential oil of Chrysanthemum morifolium-Litsea cubeba and a preparation method and application thereof, which has the following beneficial effects:

[0017] (1) The present invention provides a mixed solution of chrysanthemum essential oil and Litsea cubeba essential oil, which, after being mixed, significantly improves the activity of inhibiting Fusarium oxysporum and is used for the prevention and treatment of the pathogenic bacteria of golden thread vine stem rot.

[0018] (2) The raw materials of the present invention, Chrysanthemum officinale and Litsea cubeba, are both derived from natural plants, have rich essential oil components, and have complex and diverse antibacterial mechanisms, which can effectively solve the problem of drug resistance of pathogens.

[0019] (3) The essential oil of the present invention has the advantages of being green, safe, pollution-free and residue-free, and can replace chemical pesticides in the field of precious Chinese medicinal materials planting.

[0020] (4) The chrysanthemum seed and Litsea cubeba selected in the present invention have a wide source and a high essential oil content, and the extraction process of the present invention is simple, the extraction time is short, the energy consumption is low, and the overall cost is low, which is conducive to large-scale production and can be used for the prevention and treatment of stem rot of precious Chinese medicinal materials such as roxburghii. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on the provided drawings without paying creative work.

[0022] Figure 1 The accompanying drawing is a total ion flow diagram of chrysanthemum essential oil extracted by microwave distillation of the present invention;

[0023] Figure 2 The accompanying drawing is a total ion current diagram of the essential oil of Litsea cubeba extracted by microwave distillation of the present invention, and the unit of the abscissa is min;

[0024] Figure 3 The accompanying drawing is an antibacterial curve diagram of the Chrysanthemum Mirificum-Litsea Cubeba essential oil of the present invention. DETAILED DESCRIPTION

[0025] The following will be combined with the drawings in the embodiments of the present invention to clearly and completely describe the technical solutions in the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.

[0026] Example 1

[0027] The preparation method of chrysanthemum seed essential oil and litsea cubeba essential oil comprises the following specific steps:

[0028] The dried Lishui Suichang chrysanthemum rice and Guilin Guanyang Litsea cubeba were used as raw materials, crushed and passed through a 60-mesh sieve for later use, 50g of the raw materials were weighed and placed in a round-bottom flask, and 10 times the volume of water was added. After soaking for 0.5h, the flask was placed in a microwave extractor and irradiated and extracted for 15min under the condition of microwave power of 320W, and then the crude essential oils were collected and dehydrated with anhydrous sodium sulfate to obtain chrysanthemum rice essential oil and Litsea cubeba essential oil. The experiment was repeated 3 times, and the yield was calculated and placed in a brown bottle at 4°C for later use. The results are shown in Table 1.

[0029] Essential oil yield (%) = essential oil weight / raw material dry weight × 100%

[0030] Example 2

[0031] The preparation method of chrysanthemum seed essential oil and litsea cubeba essential oil comprises the following specific steps:

[0032] The dried Lishui Suichang chrysanthemum rice and Guilin Guanyang Litsea cubeba were used as raw materials, crushed and passed through a 60-mesh sieve for later use, 50g of the raw materials were weighed and placed in a round-bottom flask, and 20 times the volume of water was added. After soaking for 0.75h, the flask was placed in a microwave extractor and irradiated and extracted for 30min under the condition of microwave power of 480W, and then the crude essential oils were collected and dehydrated with anhydrous sodium sulfate to obtain chrysanthemum rice essential oil and Litsea cubeba essential oil. The experiment was repeated 3 times, and the yield was calculated and placed in a brown bottle at 4°C for later use. The results are shown in Table 1.

[0033] Example 3

[0034] The preparation method of chrysanthemum seed essential oil and litsea cubeba essential oil comprises the following specific steps:

[0035] The dried Lishui Suichang chrysanthemum rice and Guilin Guanyang Litsea cubeba were used as raw materials, crushed and passed through a 60-mesh sieve for later use, 50g of the raw materials were weighed and placed in a round-bottom flask, and 40 times the volume of water was added. After soaking for 1 hour, the flask was placed in a microwave extractor and irradiated and extracted for 45 minutes under the condition of microwave power of 640W, and then the crude essential oils were collected and dehydrated with anhydrous sodium sulfate to obtain chrysanthemum rice essential oil and Litsea cubeba essential oil. The experiment was repeated 3 times, and the yield was calculated and placed in a brown bottle at 4°C for later use. The results are shown in Table 1.

[0036] Table 1 Comparison of the yields of chrysanthemum essential oil and Litsea cubeba essential oil

[0037] Yield of chrysanthemum essential oil (%) Yield of essential oil from Litsea cubeba (%) Example 1 <![CDATA[2.65±0.17 b ]]> <![CDATA[2.92±0.38 c ]]> Example 2 <![CDATA[3.24±0.21 a ]]> <![CDATA[4.33±0.29 a ]]> Example 3 <![CDATA[2.82±0.15 b ]]> <![CDATA[3.68±0.21 b ]]>

[0038] The results in Table 1 show that the yields of chrysanthemum essential oil and litsea cubeba essential oil in Example 2 are 3.24% and 4.33% respectively, which are significantly higher than the yields of each essential oil in Examples 1 and 3. Therefore, Example 2 was used to prepare essential oils and conduct subsequent experiments.

[0039] Test Example 1 GC-MS Analysis and Detection of Chrysanthemum Essential Oil and Litsea Cubeba Essential Oil

[0040] The chrysanthemum essential oil and the lily seed essential oil obtained in Example 2 were analyzed by GC-MS respectively. Specifically: the chromatographic column is HP-5MS (30.0m×250μm, 0.25μm); the starting temperature is 50℃ and maintained for 2min, and then it is increased to 180℃ and maintained for 5min at a rate of 10℃ / min, and then it is increased to 300℃ and maintained for 10min at a rate of 10℃ / min; the vaporization chamber temperature is 300℃; the transmission line temperature is 300℃; the carrier gas is He; the carrier gas flow rate is 1.0mL / min; the split ratio is 100:1, and the injection volume is 0.5μL. Mass spectrometry conditions: EI source; electron energy 70eV; ion source temperature 230℃; quadrupole 150℃; scanning mode is Scan; the scanning mass range is 35~500u. Qualitative analysis: The detected components are qualitatively analyzed using the MS database NIST11 and retention time; the retention index is calculated using the normal alkane data; the column loss peak should be deducted from the database screening results. Quantitative analysis: Area normalization method is used to determine the percentage of the peak area of ​​the identified component to the sum of the areas of all identified components. According to the above method, the NIST database was searched, and the impurity peaks such as polysilane (column loss component) were deducted, and the area normalization method was used for quantification, and the retention index was calculated using the retention time of n-alkane. The results are shown in Tables 2, 3 and Figure 1 , Figure 2 .

[0041] Table 2 GC-MS analysis results of chrysanthemum essential oil

[0042]

[0043]

[0044]

[0045]

[0046] Table 3 GC-MS analysis results of Litsea cubeba essential oil

[0047]

[0048]

[0049] As shown in Tables 2 and 3, the components with a relative content of more than 0.1% in the essential oil of Chrysanthemum chrysanthemum are richer than those in the essential oil of Litsea cubeba. 74 components with a relative content of more than 0.1% were identified in the essential oil of Chrysanthemum chrysanthemum, accounting for 97.72% of the total content of the essential oil, among which the contents of monoterpenoid hydrocarbons, oxygenated monoterpenoid hydrocarbons, sesquiterpenoid hydrocarbons and oxygenated sesquiterpenoid hydrocarbons were 0.2%, 23.38%, 29.55% and 33.85% respectively, and the total amount of oxygenated terpenoid compounds could reach 57.23%, which may be the reason for its excellent antibacterial ability. In addition, the top five in relative content are (Z)-β-farnesene (13.88%), α-thujone (12.01%), T-cadinol (7.44%), (Z)-chrysanthemum chrysanthemum (6.31%) and α-gingerene (5.54%), among which chrysanthemum chrysanthemum is known as "green pesticide". Thirty-two components with a relative content of more than 0.1% were identified in the essential oil of Litsea cubeba, accounting for 94.04% of the total content of the essential oil, of which monoterpenoid hydrocarbons, oxygenated monoterpenoid hydrocarbons, sesquiterpenoid hydrocarbons and oxygenated sesquiterpenoid hydrocarbons were 75.96%, 0.58%, 6.55% and 11.22% respectively. At the same time, the main components of the essential oil of Litsea cubeba are citral and cis-citral, with contents of 36.11% and 31.16% respectively, followed by methyl heptenone (6.27%), capric acid (2.53%), lauric acid (2.43%), etc. Among them, citral is widely used in antibacterial research.

[0050] Test Example 2

[0051] The determination of the growth of Fusarium oxysporum by different volume ratios of chrysanthemum and litsea cubeba essential oils was carried out as follows:

[0052] Step 1: Prepare 40 mg / mL essential oil solution of Chrysanthemum vulgare essential oil and Litsea cubeba essential oil in different volume ratios (3:1, 2:1, 1:1, 1:2 and 1:3) with 2% DMSO, shake evenly, and filter through a 0.22 μm filter membrane for later use.

[0053] Step 2: The Fusarium oxysporum in the present invention was isolated from the stem of Anoectochilus roxburghii with stem rot, and was provided by Associate Professor Zheng Ying of the School of Food and Health of Zhejiang A&F University. 100 μL of PDB liquid culture medium was taken, and the cultured bacteria were scraped into the liquid culture medium in an appropriate amount. After diluting to a certain concentration, the concentration of the spore suspension was adjusted to 1×10 using a hemocytometer under an optical microscope. 6 cfu / mL, for future use.

[0054] Step 3: Heat and melt 20 mL of PDA medium and add 1 mL of 1×10 6 cfu / mL spore suspension, mix well and pour into the plate. After cooling, use a 6mm diameter puncher to punch a hole in the center of the 60mm plate, inject different volume ratios of chrysanthemum essential oil and Litsea cubeba essential oil into the hole, use sterile water as the blank group, and repeat each treatment three times. Incubate in a constant temperature incubator at 25℃ for about 5 days. Measure the diameter of the inhibition zone, record the data and calculate the growth inhibition rate (%) of each group. The growth inhibition rate (%) calculation formula is as follows:

[0055] Growth inhibition rate (%) = [(blank group colony diameter - experimental group colony diameter) / blank group colony diameter] × 100%

[0056] The experimental data are expressed as mean ± standard deviation (SD), and the experiment was repeated 3 times. SPSS22.0 was used for statistical analysis to analyze the differences in the mean values ​​of each group. Lowercase letters (a, b, c, d and e) indicate a significance level of 0.05, that is, there is a 5% error probability in the results. Different letters indicate significant differences between the two groups.

[0057] The results are shown in Table 4.

[0058] Table 4 Effects of different volume ratios of chrysanthemum essential oil and litsea cubeba essential oil on the growth of Fusarium oxysporum

[0059]

[0060] As shown in Table 4, after 5 days of intervention with essential oils, evaluation was performed in terms of colony diameter and growth inhibition rate, and it was found that the essential oil groups with different volume ratios all showed significant inhibition of the growth of Fusarium oxysporum, especially when the volume ratio of Chrysanthemum chrysanthemum essential oil to Litsea cubeba essential oil was 1:1, the colony diameter was the smallest, and the growth inhibition rate reached 84.06±4.90%. Therefore, the essential oil volume ratio of 1:1 was selected as the optimal composite ratio, and Chrysanthemum chrysanthemum-Litsea cubeba composite essential oil was prepared for subsequent experiments.

[0061] Test Example 3

[0062] The minimum inhibitory and bactericidal concentrations of the compound essential oil of Chrysanthemum chrysanthemi and Litsea cubeba against Fusarium oxysporum were determined in the following specific steps:

[0063] Step 1: Mix the chrysanthemum essential oil and the litsea cubeba essential oil in a volume ratio of 1:1, and prepare them into a 40 mg / mL essential oil solution with a PDB medium containing 2% DMSO, shake them evenly, and filter them through a 0.22 μm filter membrane as a comparative test group. Mix the chrysanthemum essential oil and the litsea cubeba essential oil in a volume ratio of 1:1, and prepare them into a 40 mg / mL essential oil solution with a PDB medium containing 2% DMSO, shake them evenly, and filter them through a 0.22 μm filter membrane as a test group.

[0064] Step 2: Pipette 100 μL of PDB liquid medium, scrape the cultured bacteria into the liquid medium, dilute to a certain concentration, and use a hemocytometer to adjust the concentration of the spore suspension to 1×10 6 cfu / mL, for future use.

[0065] Step 3: First, the essential oil was diluted in a 96-well plate with sterilized PDB liquid medium at a concentration of 40, 20, 10, 5, 2.5, 1.25, 0.63, 0.31, 0.16, and 0.08 mg / mL, respectively. Then, an equal volume of 1×10 6 cfu / mL bacterial suspension, with sterile water as the blank group, and then cultured in a 25℃ constant temperature incubator until the blank group was full of bacteria. Finally, the experimental results of the essential oil groups with different concentrations were read, and the lowest drug concentration corresponding to the complete inhibition of bacterial growth was regarded as the minimum inhibitory concentration. Subsequently, under sterile conditions, 50μL of the bacterial solution from the wells without bacterial growth in the 96-well plate was pipetted onto the PDA solid culture medium, evenly spread, and then cultured for 5 days. The lowest drug concentration corresponding to no colony growth was regarded as the minimum bactericidal concentration. The results are shown in Table 5.

[0066] Table 5 Minimum inhibitory and minimum bactericidal concentrations of chrysanthemum and litsea cubeba composite essential oil against Fusarium oxysporum

[0067]

[0068] As shown in Table 5, the minimum inhibitory concentration and minimum bactericidal concentration of the chrysanthemum and litsea cubeba composite essential oil against Fusarium oxysporum were 5 mg / mL and 5 mg / mL, respectively, which were significantly lower than those of the chrysanthemum essential oil and litsea cubeba essential oil groups with the same concentration. It can be seen that the antibacterial effect of the composite essential oil is more significant.

[0069] Test Example 4

[0070] Determination of the half inhibitory concentration of the compound essential oil of Chrysanthemum and Litsea cubeba (the volume ratio of Chrysanthemum essential oil to Litsea cubeba essential oil is 1:1) against Fusarium oxysporum:

[0071] Step 1: Dissolve the compound essential oil in PDB liquid culture medium containing 2% DMSO, filter through a 0.22 μm filter membrane, and set aside.

[0072] Step 2: Add the essential oil solution prepared in step 1 to the PDA culture medium to prepare a drug-containing PDA plate with a volume of 20 mL and final essential oil concentrations of 10, 5, 2.5, 1.25, 0.625, and 0.3125 mg / mL, respectively, and pour it into a culture dish with a diameter of 60 mm. The PDA plate with sterile water added is used as the blank group. After the plate is condensed, the Fusarium oxysporum cake (d = 6 mm) is inverted in the center of the drug-containing PDA plate and cultured in a constant temperature incubator at 25°C for about 5 days. The colony diameter of each treatment was measured by the cross method, and the growth inhibition rate and IC of each group were calculated according to Formula 1. 50 The results are shown in Table 6.

[0073] Table 6 The half inhibitory concentration of the compound essential oil of chrysanthemum and litsea cubeba against Fusarium oxysporum

[0074]

[0075] As shown in Table 6 and Figure 3 As shown in the figure, the half inhibitory concentration of the composite essential oil of Chrysanthemum chrysanthemum and Litsea cubeba against Fusarium oxysporum was 0.27±0.05 mg / mL, which was significantly lower than 0.43±0.12 mg / mL and 1.17±0.18 mg / mL of Chrysanthemum chrysanthemum essential oil and Litsea cubeba essential oil, which once again showed that the antibacterial effect of the composite essential oil was more significant.

[0076] Test Example 5

[0077] Determination of the antibacterial zone of the compound essential oil of Chrysanthemum villosum and Litsea cube

[0078] The antibacterial activity of essential oils against Fusarium oxysporum was determined by the filter paper method. Pour the PDA solid medium into a sterile culture dish while it is still hot. After the medium cools and solidifies, use a pipette to take 0.2 mL of bacterial solution (1×10 6 cfu / mL) onto the surface of the solid culture medium and spread it evenly on the surface with a sterilized coating rod. Take 10 μL of chrysanthemum essential oil, Litsea cubeba essential oil (the essential oil prepared in Example 2), chrysanthemum-Litsea cubeba composite essential oil (v:v=1:1) and 10 μL of 2% DMSO respectively, prepare 3 6mm filter paper pieces for each group and soak them in the above solution, then take them out and stick them on the culture medium coated with the bacterial solution, and culture them in a constant temperature incubator at 25°C for about 5 days. After the culture is completed, record the diameter of the inhibition zone. The results are shown in Table 7. The filter paper piece with 2% DMSO is the negative control group.

[0079] Table 7 Inhibition zone diameter of chrysanthemum and litsea cubeba composite essential oil against Fusarium oxysporum

[0080]

[0081] As shown in Table 7, the antibacterial zone of the chrysanthemum-litsea cubeba composite essential oil is 18.2±0.4 mm, which is much larger than that of chrysanthemum essential oil and litsea cubeba essential oil used alone, which once again proves the antibacterial effect of the chrysanthemum-litsea cubeba composite essential oil.

[0082] The above description of the disclosed embodiments enables one skilled in the art to implement or use the present invention. Various modifications to these embodiments will be apparent to one skilled in the art, and the general principles defined herein may be implemented in other embodiments without departing from the spirit or scope of the present invention. Therefore, the present invention will not be limited to the embodiments shown herein, but rather to the widest scope consistent with the principles and novel features disclosed herein.

Claims

1. A composite essential oil of Chrysanthemum officinale and Litsea cubeba, characterized in that: The compound is prepared from chrysanthemum essential oil and litsea cubeba essential oil in a volume ratio of (1-3):(1-3).

2. A chrysanthemum seed-litsea cubeba composite essential oil according to claim 1, characterized in that The preparation methods of the chrysanthemum seed essential oil and the litsea cubeba essential oil are as follows: The dried chrysanthemum and litsea cubeba are respectively used as raw materials, which are crushed and sieved for later use; the crushed and sieved raw materials are respectively weighed and placed in round-bottom flasks, and 10 to 40 times the volume of water is added, and after soaking for 0.5 to 1 hour, the flask is placed in a microwave extractor, and irradiated and extracted for 15 to 45 minutes under the condition of microwave power of 320 to 640 W, and the extracted oil-water mixture is collected and dried and dehydrated to obtain chrysanthemum essential oil and litsea cubeba essential oil, respectively.

3. A method for preparing a composite essential oil of Chrysanthemum officinale and Litsea cubeba, characterized in that: The specific steps are as follows: (1) using the method according to claim 2 to prepare chrysanthemum essential oil and litsea cubeba essential oil respectively; (2) The prepared Chrysanthemum chrysanthemum essential oil and Litsea cubeba essential oil are mixed in a volume ratio of (1-3): (1-3) to obtain Chrysanthemum chrysanthemum-Litsea cubeba composite essential oil.

4. Use of the chrysanthemum seed-litsea cubeba composite essential oil according to claim 1 or 2 in preventing and treating roxburghii roxburghii stem rot.

5. Use of the composite essential oil of Chrysanthemum vulgare and Litsea cubeba according to claim 1 or 2 in inhibiting Fusarium oxysporum.

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