CN115039797A - Clove essential oil antibacterial mixed solution and application thereof - Google Patents
Clove essential oil antibacterial mixed solution and application thereof Download PDFInfo
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- CN115039797A CN115039797A CN202210296159.7A CN202210296159A CN115039797A CN 115039797 A CN115039797 A CN 115039797A CN 202210296159 A CN202210296159 A CN 202210296159A CN 115039797 A CN115039797 A CN 115039797A
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- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims abstract description 20
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N65/00—Biocides, pest repellants or attractants, or plant growth regulators containing material from algae, lichens, bryophyta, multi-cellular fungi or plants, or extracts thereof
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/28—Myrtaceae [Myrtle family], e.g. teatree or clove
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- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Engineering & Computer Science (AREA)
- Plant Pathology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Mycology (AREA)
- Agronomy & Crop Science (AREA)
- Dentistry (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Botany (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses clove essential oil antibacterial mixed liquid and application thereof, and belongs to the technical field of biological control. The clove essential oil bacteriostatic mixed solution comprises the following components in percentage by mass: 1-2 per mill of clove essential oil; 5-10% of alcohol solvent; 0.5-1 per mill of cosolvent; the balance being water. Wherein the alcohol solvent comprises one or more of propylene glycol, ethanol and butanediol; the cosolvent is Tween 80. The clove essential oil can effectively inhibit the growth of fusarium oxysporum hyphae and the germination of fusarium oxysporum spores. The bacteriostatic mixed liquid containing the clove essential oil prepared by the invention can effectively inhibit the activity of fusarium oxysporum and is used as a bactericide for preventing and treating fusarium oxysporum. The invention expands a new idea of pollution-free control based on the characteristic that plants have antibacterial activity.
Description
The technical field is as follows:
the invention belongs to the technical field of biological control, and particularly relates to clove essential oil antibacterial mixed liquid and application thereof.
Background art:
bananas (Musa nanaLour.) belongs to the family Musaceae (Musaceae) Musa genus (A)Musa) The fruit is originally produced in tropical and subtropical regions, is cultivated in Guangdong, Guangxi, Fujian, Taiwan, Yunnan and Hainan, is one of four fruits in the world, has the yield second to that of apples, oranges and pears, and is also defined as the fourth grain crop by the grain agriculture organization of the United nations. The banana vascular wilt is the first killer of banana production, and the pathogenic bacteria is fusarium oxysporum Guba specialization type 2Fusarium oxyporum f.sp.cubense(FOC)）]Is a devastating soil-borne fungal disease which causes necrosis of banana vascular bundles. The disease transmission path is wide, the disease can be transmitted through media such as water, soil and the like, and once the banana garden is infected with bacteria, the banana garden can be attacked by garden damage if no measures are taken. Research has found that leek extract can inhibit spore germination by affecting the vegetative metabolism of fusarium oxysporum spores; the artemisia annua leaf methanol extract can reduce the spore germination rate and the spore activity of fusarium oxysporum.
The clove is various in variety and wide in planting area in China, and besides being used as daily ornamental flower, the clove essential oil product is widely applied to industries such as food and medicine due to the characteristics of no pollution, safety and bacteriostasis. The clove essential oil is natural vegetable oil extracted from flower buds, stems or leaves of clove plants, mainly comprises eugenol, acetyl eugenol, beta-caryophyllene, methyl salicylate and the like, wherein the eugenol is considered as a main effective component for bacteriostasis. The clove essential oil has high-efficiency bacteriostatic activity, and researches show that the growth inhibition time of the clove essential oil on penicillium and botrytis cinerea can reach 9 days and 8 days.
Therefore, the development of a method for inhibiting fusarium oxysporum by using clove essential oil is a new way for biologically preventing and treating banana vascular wilt and develops a nuisanceless prevention and treatment idea.
Disclosure of Invention
The invention aims to provide clove essential oil bacteriostatic mixed liquid and application thereof.
The clove essential oil bacteriostatic mixed solution comprises the following components in percentage by mass, wherein the sum of the mass percentages is 100 percent: 1-2 per mill of clove essential oil; 5-10% of alcohol solvent; 0.5-1 per mill of cosolvent; the balance being water. The alcohol solvent comprises one or more of propylene glycol, ethanol and butanediol; the cosolvent is tween 80.
(2) The preparation method of the clove essential oil bacteriostatic mixed solution comprises the following steps:
mixing an alcohol solvent and clove essential oil, adding a cosolvent, adding water, and independently stirring the mixed solution at 50-100 rpm/min for 5-10 min to prepare the clove essential oil antibacterial mixed solution.
A bactericide containing the above flos Caryophylli essential oil antibacterial mixed solution is provided.
The clove essential oil bacteriostatic mixed solution is applied to preventing and treating fusarium oxysporum.
The bactericide is applied to the prevention and treatment of fusarium oxysporum.
The invention has the advantages that: the invention provides clove essential oil bacteriostatic mixed liquid, wherein the clove essential oil can effectively inhibit the growth of fusarium oxysporum hyphae and the germination of fusarium oxysporum spores. The bacteriostatic mixed liquid containing the clove essential oil prepared by the invention can effectively inhibit the activity of fusarium oxysporum and is used as a bactericide for preventing and treating fusarium oxysporum. The invention expands the idea of pollution-free control based on the characteristic that plants have bacteriostatic activity.
Description of the drawings:
FIG. 1 shows the inhibition of the growth of No. 4 hyphae of physiological race of Fusarium oxysporum by clove essential oil with different concentrations.
FIG. 2 is the effect of clove essential oil on the germination of Fusarium oxysporum physiological race No. 4 spores.
Detailed Description
The present invention is further described in detail below with reference to specific examples, which are provided to assist those skilled in the art in further understanding the present invention, but are not intended to limit the present invention in any way.
Example 1: indoor bacteriostasis rate determination and results
Fusarium oxysporum physiological race No. 4 used in this experimentFusarium oxyporum f.sp.cubense （FOC4）]The strain is stored at 4 deg.C in laboratory, and the culture medium is potato agar glucose culture medium (PDA for short) by hypha growth method. The PDA culture medium formula comprises: 200 g of potato (peeled), 20g of glucose, 15-20g of agar and 1000mL of distilled water, and the pH value is natural.
The preparation method of the PDA culture medium comprises the following steps: cleaning and peeling 200 g of potatoes, cutting the potatoes into slices, adding water, boiling for 20-30 minutes, filtering the potatoes in a beaker by using 4 layers of gauze, fixing the volume to 1000mL, adding 15-20g of agar, adding 20g of glucose, stirring to prevent sticking, subpackaging the potatoes in 250 mL triangular bottles after boiling, packaging the potatoes in 200 mL triangular bottles with 150-.
The experimental method comprises the following steps: by hypha growth rate method
1. Firstly, the physiological race 4 of fusarium oxysporum (FOC 4) is inoculated on a PDA plate and cultured for 5 to 7 days at the temperature of between 25 and 28 ℃.
2. Heating PDA culture medium to melt, cooling to 45-50 deg.C, adding flos Caryophylli essential oil with different concentrations into sterile culture dish in ultra-clean bench, adding 25 mL PDA culture medium, and mixing by left and right pushing to obtain flat plate with medicine.
3. Under the aseptic condition in an ultra-clean workbench, a hole puncher with the diameter of 8mm is used for punching a circular fungus cake on the edge of FOC4 hypha cultured for 5-7 days, then a inoculating needle is used for inoculating the circular fungus cake to the center of a PDA (personal digital assistant) plate containing medicine, and then a culture dish is placed upside down in an incubator at the temperature of 25-28 ℃ for culture. Experimental group prepared different concentrations of clove essential oil drug-containing plates, 0.02 uL/mL, 0.04 uL/mL, 0.06 uL/mL, 0.08 uL/mL, 0.10 uL/mL, 0.12 uL/mL, continuously observed for 6 days and measured for hypha diameter. After the blank control group CK grows over the flat plate (about 7 days), observing the growth condition of hyphae (see figure 1), measuring the diameter by adopting a cross method, counting data, and calculating the inhibition rate. Each treatment was repeated 3 times.
Control hyphal diameter D 0 Diameter D of hypha on the drug-containing plate 1 ,
Inhibition ratio (%) [ (D) 0 -0.8）-（D 1 -0.8）]/（D 0 -0.8）× 100%
4. The results of the activity assay of plant pathogenic fungi are shown in Table 1.
TABLE 1 inhibition rate of clove essential oil on Fusarium oxysporum physiological race No. 4
Note that 3 replicates were used for each treatment and the data in the table are the average of the 3 replicates.
Example 2: method for measuring influence of clove essential oil on spore germination of fusarium oxysporum physiological race No. 4 (FOC 4) by using pendant drop method
1. Inoculating FOC4 test strain in potato glucose agar (PDA) culture medium, culturing at constant temperature of 25-28 deg.C in light incubator for 10-14d until the powder is seen, indicating that conidia are generated in large amount, eluting with sterile water, and placing in sterile culture tube.
2. Prepared with sterile water to a concentration of 1 × 10 6 cfu/mL spore suspension.
3. Dripping clove essential oil with different concentrations into spore suspension, respectively adding 0.5 uL, 1.0 uL, 1.5 uL and 2.0 uL of clove essential oil into 500uL of spore suspension, stirring and mixing uniformly, dripping on a concave slide groove, inversely placing in a large culture dish, placing water-absorbing sterile filter paper in the culture dish, and performing moisturizing culture. Incubate at constant temperature illumination 25-28 ℃ with 3 slides observed each time as replicates, 3 fields observed per slide.
Conidiophores germination rate% = number of germinated spores/total number of spores × 100%
4. The effect of clove essential oil on the germination of Fusarium oxysporum physiological race No. 4 spores is shown in figure 2, and conidia are shriveled and fine.
5. The test observation and record show that the germination rate of conidia after the clove essential oil is treated is zero.
TABLE 2 conidium germination rate after treatment with clove essential oil
Example 3: fusarium oxysporum inhibiting effect by flos Caryophylli mixed solution
1. Test materials: 4-6 leaf banana (Brazil banana) plantlets were taken as the test.
2. Preparation of pathogen spore suspension: transferring a test strain Fusarium oxysporum physiological race No. 4 (FOC 4) to a potato glucose culture medium (PDA) to activate for 5-7 d at 28 ℃, picking small hyphae to inoculate in 150 mL sterile potato glucose culture solution (PDB), carrying out shake culture for 7 d in a shaking table with constant temperature of 26 ℃ and 110r/min, filtering out hyphae from the bacterial solution by two layers of sterilized gauze, centrifuging the filtrate for 20 min at 4000r/min, discarding the supernatant, and washing out the precipitated spores by sterile water to prepare a mother solution. Calculating spore concentration with blood counting plate, and adjusting mother liquor to 1 × 10 with sterile water 6 cfu/mL of spore suspension for use in the inoculation experiment.
3. The inoculation method adopts a root soaking method for inoculation: immersing root system of 4-6 leaf banana (Brazil banana) plantlet into the above 1 × 10 6 cfu/mL of spore suspension of Fusarium oxysporum physiological race No. 4 (FOC 4) for 20 min, and then transplanted into a plastic cup containing sterilized soil with clear water as a control.
4. Treatment group: taking 100mL of clove essential oil bacteriostatic mixed solution to irrigate roots to be applied to inoculated 4-6-leaf banana (Brazil banana) seedlings, and irrigating roots at intervals of 2d for 1 time by using 100mL of clove essential oil bacteriostatic mixed solution.
The clove essential oil bacteriostatic mixed solution comprises the following components in percentage by mass of 100 percent: 1-2 per mill of clove essential oil, 5-10 percent of alcohol solvent, 0.5-1 per mill of cosolvent and the balance of water. The alcohol solvent can be one or more of propylene glycol, ethanol and butanediol. The cosolvent comprises tween 80.
The preparation of the clove essential oil antibacterial mixed solution comprises the following steps: mixing an alcohol solvent and clove essential oil, adding a cosolvent, adding water, and independently stirring the mixed solution at 50-100 rpm/min for 5-10 min to prepare the antibacterial mixed solution.
Treatment 1: the formula for preparing 1L of clove essential oil mixed liquid is as follows: 1 mL of clove essential oil, 50mL of 60vol% ethanol, 800.8 mL of tween and 1L of water.
And (3) treatment 2: the formula for preparing 1L of clove essential oil mixed liquid is as follows: 1 mL of clove essential oil, 50mL of propylene glycol, 800.8 mL of tween and about 1L of water.
And (3) treatment: the formula for preparing 1L of clove essential oil mixed liquid is as follows: 1 mL of clove essential oil, 50mL of butanediol, 800.8 mL of tween and about 1L of water.
100mL of the clove essential oil mixed solution prepared by each treatment is applied to inoculated 4-6-leaf banana (Brazil banana) seedlings by root irrigation, and 100mL of the clove essential oil bacteriostatic mixed solution prepared by each treatment is applied for 1 time at intervals of 2d root irrigation. And then, pouring clear water to supplement the water required by the growth of the bananas, and surveying and counting the morbidity and the relative control effect after 20 days.
Incidence (%) = number of diseased plants/total number of investigated plants × 100
Relative control effect% = (inoculation positive control morbidity-treatment group morbidity)/inoculation positive control morbidity x 100
5. Statistics of morbidity of bacteriostatic mixed solution
Claims (5)
1. The clove essential oil antibacterial mixed liquid is characterized in that: the clove essential oil bacteriostatic mixed solution comprises the following components in percentage by mass of 100 percent: 1-2 per mill of clove essential oil; 5-10% of alcohol solvent; 0.5-1 per mill of cosolvent; the balance being water.
2. The clove essential oil bacteriostatic mixed liquor as claimed in claim 1, wherein: the alcohol solvent comprises one or more of propylene glycol, ethanol and butanediol; the cosolvent is Tween 80.
3. A bactericide containing the clove essential oil bacteriostatic mixture solution of claim 1.
4. The application of the clove essential oil bacteriostatic mixed solution in the prevention and treatment of fusarium oxysporum as claimed in claim 1.
5. Use of the fungicide according to claim 3 for the control of fusarium oxysporum.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117617272A (en) * | 2023-11-29 | 2024-03-01 | 中国热带农业科学院热带生物技术研究所 | Application of plant essential oil in preventing and controlling banana pathogenic fungi |
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| CN109452320A (en) * | 2018-11-15 | 2019-03-12 | 云南中医学院 | The purposes of Fructus Caryophylli volatile oil |
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