CN109452320A - The purposes of Fructus Caryophylli volatile oil - Google Patents
The purposes of Fructus Caryophylli volatile oil Download PDFInfo
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- CN109452320A CN109452320A CN201811359510.2A CN201811359510A CN109452320A CN 109452320 A CN109452320 A CN 109452320A CN 201811359510 A CN201811359510 A CN 201811359510A CN 109452320 A CN109452320 A CN 109452320A
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- volatile oil
- fructus caryophylli
- pathogenic bacteria
- caryophylli volatile
- hymexazol
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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
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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
- A01N65/08—Magnoliopsida [dicotyledons]
- A01N65/28—Myrtaceae [Myrtle family], e.g. teatree or clove
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- Agronomy & Crop Science (AREA)
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- Natural Medicines & Medicinal Plants (AREA)
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Abstract
The invention discloses a kind of purposes of Fructus Caryophylli volatile oil；I.e. it is agriculturally preventing and treating the application in Chinese medicine or crop root rot or the application in the pathogenic bacteria that prevention and treatment causes Chinese medicine or crop overground part that disease occurs；The experimental results showed that Fructus Caryophylli volatile oil can inhibit 4 kinds of main pathogenic bacteria Fusarium oxysporums for causing notoginseng root rot to occur, Fusarium solani, the mycelia growth that rotten mould and breaking-up column spore is mould, and the mycelia growth that 3 kinds of pathogenic bacteria for causing Radix Notoginseng overground part disease to occur inhibit botrytis cinerea, colletotrichum gloeosporioides Penz, Rhizoctonia solani Kuhn can be completely inhibited；Fructus Caryophylli volatile oil fungistatic effect is suitable with chemical pesticide hymexazol fungistatic effect, and Fructus Caryophylli is compounded with hymexazol all has synergistic effect to multiple bacterial strains；Due to the effumability of volatile oil, and have many advantages, such as fragrance keep away it is dirty, therefore with its exploitation chemical fertilizer or pesticide will have safely, effectively, low-residual.
Description
Technical field
The present invention relates to the purposes of Fructus Caryophylli volatile oil more particularly to Fructus Caryophylli volatile oil to Chinese medicine or crop root rot
And the preventive and therapeutic effect to Chinese medicine or crop overground part multiple diseases, belong to biological pesticide technical field.
Background technique
Radix Notoginseng (Panax notoginseng(Burk.) F.H.Chen) also known as pseudo-ginseng, it is Araliaceae Panax many years
Raw herbaceous plant has dissipating stasis and stanching bleeding, detumescence ding-tong and other effects, is the distinctive rare cultivation medicinal plant of China, main product
Southwestern China area, wherein in Wenshan Prefecture, Yunnan Province away from modern existing more than 400 years cultivation histories.Roots of Panax Notoginseng, stem, leaf, Hua Junke
It is used as medicine, scope of medication is very extensive, and social required quantity is being continuously increased, with increasing sharply for notoginseng planting area, disease
Problem is got worse, it has also become the major obstacle of Radix Notoginseng production.The cultivation of most of Chinese herbal medicines all has genuineness, by geology soil
The influence of earth characteristic, weather conditions and the plantation factors such as tradition, thus inevitably there is a large amount of continuous cropping, cause yield and
Quality reduces, drug effect decline, it has also become urgent problem to be solved in agricultural production.
Currently, various chemical agents are relied primarily on to the prevention and treatment of Radix Notoginseng disease, though chemical pesticide effectively, considerably increases
The input cost of peasant more seriously polluted soil and reduce medical material quanlity.In modern agricultural production, chemical pesticide is wide
Model is applied to the prevention and treatment of pest and disease damage, is the main means of current control of plant disease.However, its fatal defects are in useization
There are problems that " 3R " (residual, drug resistance and regeneration) when learning sterilization dose, insecticide, the pesticide of high residue will cause serious ring
Border pollution.For example, hymexazol is a kind of new pesticide, prevention of disease and control have a unique effect, however it is to skin
Skin and eyes are irritant, can cause hepatotoxicity wind agitation.
Plant volatile oil is not limited only to traditional medicine, food, tobacco, alcohol and tobacco, it may also be used for plant protection (is killed
Worm, antibacterial, weeding etc.), this opens up a new way without the utilization suspected of plant ecosystem.Plant volatile oil is as plant
Material resource pesticides application is consistent with the public nuisance-free agricultural chemicals theory of suggestion in plant protection art.In this sense, phytovolatilization
Oil can also be classified as third generation pesticide.
Fructus Caryophylli is the almost ripe fruit of plant clove of myrtaceae (medicinal cloves), and dry fruit is in oval or ellipse
Shape；It is Fructus Caryophylli acrid flavour, warm-natured；Sudden and violent heart pain can be controlled, stomach cold is vomitted, and inverse, air-cooled dentalgia, pingival atrophy, halitosis, married woman be sombre, children's hernia
Deng.
Currently, having no that document relevant to technical solution of the present invention discloses.
Summary of the invention
It is an object of that present invention to provide a kind of new applications of Fructus Caryophylli volatile oil, i.e., it is in prevention and treatment Chinese medicine or crop root-rot
Application in disease.
Fructus Caryophylli volatile oil of the present invention, which also can be applicable to prevention and treatment, causes Chinese medicine or crop overground part that causing a disease for disease occurs
In bacterium；The pathogenic bacteria be botrytis cinerea (Botrytis cinerea), colletotrichum gloeosporioides Penz (Colletotrichum gloeosporioides), Rhizoctonia solani Kuhn (Rhizoctonia solani) etc..
The purposes of Fructus Caryophylli volatile oil of the present invention is that Fructus Caryophylli volatile oil is used as to preparation prevention and treatment Chinese medicine or work
Object root rot or prevention and treatment cause the preparation of the pathogenic bacteria of Chinese medicine or crop overground part generation disease, i.e., with Fructus Caryophylli volatile oil
For active constituent, the application in the bacteria preparation that causes a disease is prevented and treated in preparation.
The ingredient (or effective component) of the present invention for preventing and treating the bacteria preparation that causes a disease is Fructus Caryophylli volatile oil, can also be added one
Acceptable auxiliary material in kind or plurality of Chinese and crop disease control preparation, or play and cooperate with other active components compounding
Bacteriostasis.
Fructus Caryophylli volatile oil can be prepared into the pesticide of various dosage forms or the chemical fertilizer of various forms by prior art device.
Fructus Caryophylli volatile oil is extracted by the conventional steam way of distillation in the present invention, anhydrous sodium sulfate drying and volatilizing oil and
It is made, the preparation method is simple, and required reagent cost is low, is conducive to mass production.
Fructus Caryophylli volatile oil can reduce plant disease incidence and the state of an illness by inhibiting pathogen spore germination and mycelia growth
Index；By with chemical pesticide synergy, achieve the purpose that reduce chemical pesticide application.
Fructus Caryophylli volatile oil is as a kind of natural Fungicidal active substance, its activity disappears or by microorganism point after use
Solution, no pollution to the environment, to safety of human and livestock；Since Fructus Caryophylli volatile oil is Extracts from Plant Recourses, effective component is complicated various,
Be the mechanism of action of multicomponent multiple target point to pathogen so that target organisms be not likely to produce several ingredients of certain or certain are generated it is anti-
Pharmacological property, to obtain lasting control efficiency.Compared with chemical synthetic pesticide, the effective component of Fructus Caryophylli volatile oil is volatile,
It is degradable, it is antibacterial without sterilizing, therefore be not easy to cause damages to other beneficial microbes, improve soil micro-ecosystem structure, promotees
It into plant and microorganism coevolution, preserves the ecological environment, improves the quality of Chinese medicine or crop.
The Fructus Caryophylli volatile oil solution of 50mg/mL is prepared, solvent is DMSO and volumetric concentration containing volumetric concentration 1%
The aqueous solution of 0.1% Tween 80,4 kinds of main pathogenic bacteria point spore reaping hooks that notoginseng root rot occurs for test Fructus Caryophylli volatile oil
Bacterium (Fusarium oxysporum), Fusarium solani (Fusarium solani), it is rotten it is mould (Pythium aphanidermatum) and damage column spore it is mould (Cylindrocarpon destructans) effect, and with leading to Radix Notoginseng
Top disease occur 3 kinds of pathogenic bacteria botrytis cinereas (Botrytis cinerea), colletotrichum gloeosporioides Penz (Colletotrichum gloeosporioides), Rhizoctonia solani Kuhn (Rhizoctonia solani) mycelia growth effect；
The present invention opens the new method for reducing chemical pesticide application, facilitates scientific worker and grinds to the volatile oil mechanism of action
Study carefully, there is broader prospect in agricultural production；The Fructus Caryophylli volatile oil of the present invention for reducing the generation of Radix Notoginseng disease, which has, to be thrown
The characteristics of entering low, easy to operate, high-efficient, volatile, less toxic, low-residual.The overall background of organic agriculture is greatly developed in country
Under, Fructus Caryophylli hair oil is the more satisfactory chemical agent substitute product for overcoming Radix Notoginseng disease, is had to the sustainable development of agricultural
Important meaning.
Detailed description of the invention
Fig. 1 be in the present invention Fructus Caryophylli volatile oil to Fusarium oxysporum (a), Fusarium solani (b), damage column spore mould (c),
The influence (colony diameter result) of rotten mould (d), grey mold (e), colletotrichum gloeosporioides Penz (f) and the growth of Rhizoctonia solani Kuhn (g) bacterium colony；
Fig. 2 be in the present invention Fructus Caryophylli volatile oil to Fusarium oxysporum, Fusarium solani, damage that column spore is mould, rotten mould, grey mold, glue
The influence (bacteriostasis rate) of spore anthrax-bacilus and the growth of Rhizoctonia solani Kuhn bacterium colony.
Specific embodiment
Below by drawings and examples, invention is further described in detail, but the scope of the present invention is not limited to
The content, method is conventional method unless otherwise specified in embodiment, and the reagent used is routine unless otherwise specified
Commercial reagent or the reagent prepared according to a conventional method.
Embodiment 1: the preparation of Fructus Caryophylli volatile oil
(1) water that 8 times of its quality is added in the Fructus Caryophylli powder that 1kg is crushed, impregnates 2h, by the way of heating distillation, benefit
With the volatile oil in steam extraction Fructus Caryophylli, extraction time 6h, condensation collects and slips out liquid；
(2) volatile oil of collection is dry with anhydrous sodium sulfate, institute's water content is removed, puts brown sample bottle into, is sealed, -20
It DEG C saves stand-by.
Embodiment 2: the activation of pathogenic bacteria
(1) be isolated from the pathogenic bacteria of Panax notoginseng root and overground part, Purification be Fusarium oxysporum (Fusarium oxysporum), Fusarium solani (Fusarium solani), it is rotten it is mould (Pythium aphanidermatum), damage column spore
Mould (Cylindrocarpon destructans), botrytis cinerea (Botrytis cinerea), colletotrichum gloeosporioides Penz
(Colletotrichum gloeosporioides) and Rhizoctonia solani Kuhn (Rhizoctonia solani) strain be seeded in
It is activated in PDA culture medium, after inoculation 4 times, takes eugonic bacterial strain spare；
(2) potato dextrose agar (PDA): potato 200g, glucose 20g, 20 g of fine jade, 1000 mL of distilled water.
The chemical constituent of embodiment 3:GC-MS measurement Fructus Caryophylli volatile oil
(1) GC condition: instrument model: Agilengt Technologies 7890B-5977B；Chromatographic column: HP-5MS 30m ×
250μm×0.25μm；
(2) source EI source temperature: 230 DEG C；Level four bars temperature: 150 DEG C；Scanning range: 30 ~ 500m/z；Injector temperature:
285℃；Split ratio 10: 1；Sample volume: 1 μ l；Electron energy: 70eV；Column oven temperature program: 50 DEG C of holding 4min, 5 DEG C/
Min is warming up to 120 DEG C, and 1 DEG C/min is warming up to 180 DEG C, and 10 DEG C/min is warming up to 280 DEG C of holding 16min；Wherein RI value root
It is calculated according to the retention time of n-alkane continuous carbon (C9-C25), by the retention time of the compound and mass spectrum and NIST
17.L database carries out comparing, determines final compound, testing result see the table below；
In table: a: retention time；B: relative peak area；± relative deviation；C: retention index.
Embodiment 4: the influence that measurement Fructus Caryophylli volatile oil grows 7 kinds of pathogenic bacteria mycelia in embodiment 2
(1) under aseptic technique, 15 mL PDA culture mediums are poured into each culture dish, after solidification to be cooled, take above-mentioned reality
The pathogenic bacteria for cultivating 7 d in example 2 in PDA culture medium are applied, is beaten with the punch of diameter 5mm along colony edge and takes fungus block, are inoculated with
In culture medium central；
(2) 4 sterile Oxford cups are equidistantly placed at 25 mm of fungi block；
(3) Fructus Caryophylli the volatile oil DMSO containing volumetric concentration 1% and volumetric concentration 0.1% by above-described embodiment 1 after dry
Tween 80 aqueous dissolution, be made concentration be 50 mg/mL Fructus Caryophylli volatile oil liquid；
(4) the 50mg/mL Fructus Caryophylli volatile oil of 200 μ L steps (3) is added (in advance with 0.22 μm of organic system filter in each Oxford cup
Filtration sterilization), 28 DEG C of constant temperature incubations in microbiological incubator, the DMSO containing volumetric concentration 1% and volumetric concentration 0.1%
Tween 80 aqueous solution and hymexazol be control, each processing sets 5 repetitions；
(5) after cultivating 7d in incubator, colony diameter is measured using " crossing method ".
The result is shown in Figure 1,2, it can be seen that Fructus Caryophylli volatile oil has obviously the mycelia growth of 7 kinds of pathogenic bacteria from Fig. 1,2
Inhibiting effect, and it is suitable with the effect of chemical agent hymexazol, wherein to the fungistatic effect of Fusarium solani significantly better than disliking
Mould spirit.
Embodiment 5: Fructus Caryophylli volatile oil MIC measurement
(1) tween of Fructus Caryophylli the volatile oil DMSO containing volumetric concentration 1% and volumetric concentration 0.1% after embodiment 1 is dry
80 aqueous dissolution, the multiple concentration gradient liquid for being 150-0.15mg/mL with doubling dilution configuration concentration range are positive
Medicine hymexazol, monomeric compound eugenol, carypohyllene and eugenol acetate are configured to concentration range with same method and are
The concentration gradient of 2.5-0.005mg/mL；
(2) pathogen that 7d is grown in embodiment 2, is rinsed with 20 mL 1/4PDA fluid nutrient mediums (not adding agar) and is cultivated
Base, it is 1 × 10 that spore concentration, which is made,4The spore suspension of a/mL；
(3) it is added in 96 orifice plates in the 50 μ L Fructus Caryophylli volatile oil and 150 μ L steps (2) of 0.22 μm of filtering with microporous membrane degerming
Manufactured spore suspension；
(4) negative control be 50 μ L contain the DMSO of volumetric concentration 1% and the Tween 80 of volumetric concentration 0.1% aqueous solution and
Manufactured spore suspension in 150 μ L steps (2)；Positive control is manufactured spore suspension in 150 μ L steps (2)；
(5) 96 28 DEG C of orifice plate constant temperature incubation 36h, then fungi growing state in inspection hole；Microplate reader is used at 595 nm
(Thermo 1510) measures the absorbance in each hole, and the corresponding drug concentration in that hole that absorbance is 0 is minimum antibacterial dense
Spend MIC.
As a result it see the table below；
Note: with a line, different letters indicate that different disposal has significant difference (P < 0.05).
Embodiment 6: Fructus Caryophylli volatile oil and hymexazol interaction measure
(1) MIC according to embodiment 5 obtained Fructus Caryophylli volatile oil and hymexazol, being configured to maximum concentration is 8 times of MIC, then
8 dilution gradients are configured using doubling dilution；
(2) by the combination addition of the hymexazol (25 μ L) of the Fructus Caryophylli volatile oil of various concentration (25 μ L) and various concentration in 96 orifice plates
It is interior, manufactured spore suspension (150 μ L) in 5 step of embodiment (2) is then added in each hole；
Manufactured spore suspension and DMSO and volumetric concentration containing volumetric concentration 1% in 5 step (2) of (3) 150 μ L embodiment
The aqueous solution of 0.1% Tween 80 is negative control；
(4) 96 orifice plates save 36h in 28 DEG C of constant temperature, and with microplate reader (Thermo 1510) colorimetric at 595nm, it is raw to detect fungi
Long situation；
(5) corresponding drug concentration the MIC measurement of drug combination: is grown as Fructus Caryophylli volatile oil and hymexazol using no fungi hole
Associated with MIC；
(6) FIC is calculated: the MIC of MIC/ volatile oil associated with FIC=volatile oil of volatile oil and hymexazol, the FIC of hymexazol=wave
The MIC of MIC/ hymexazol associated with hair oil and hymexazol；
The FIC of FICI=volatile oil FIC+ hymexazol, wherein FICI≤0.5 is synergistic effect, and 0.5 < FICI≤1 is that adduction is made
With 1<FICI≤4 are unrelated effect, and FICI>4 are antagonism；As a result it see the table below；
。
Claims (3)
1. application of the Fructus Caryophylli volatile oil in prevention and treatment Chinese medicine or crop root rot.
2. application of the Fructus Caryophylli volatile oil in the pathogenic bacteria that prevention and treatment causes Chinese medicine or crop overground part that disease occurs.
3. application according to claim 2, it is characterised in that: pathogenic bacteria be botrytis cinerea (Botrytis cinerea), glue
Spore anthrax-bacilus (Colletotrichum gloeosporioides), Rhizoctonia solani Kuhn (Rhizoctonia solani).
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| CN201811359510.2A CN109452320B (en) | 2018-11-15 | 2018-11-15 | Application of clove volatile oil |
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| CN201811359510.2A CN109452320B (en) | 2018-11-15 | 2018-11-15 | Application of clove volatile oil |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110250216A (en) * | 2019-08-05 | 2019-09-20 | 云南中医药大学 | The new application of Herba laggera volatile oil |
| CN115039797A (en) * | 2022-03-24 | 2022-09-13 | 福建省农业科学院亚热带农业研究所（福建省农业科学院蔗麻研究中心） | Clove essential oil antibacterial mixed solution and application thereof |
| 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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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110250216A (en) * | 2019-08-05 | 2019-09-20 | 云南中医药大学 | The new application of Herba laggera volatile oil |
| CN115039797A (en) * | 2022-03-24 | 2022-09-13 | 福建省农业科学院亚热带农业研究所（福建省农业科学院蔗麻研究中心） | Clove essential oil antibacterial mixed solution and application thereof |
| 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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