CN109247347A - The purposes of Fructus Citri Sarcodactylis volatile oil - Google Patents
The purposes of Fructus Citri Sarcodactylis volatile oil Download PDFInfo
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- CN109247347A CN109247347A CN201811359515.5A CN201811359515A CN109247347A CN 109247347 A CN109247347 A CN 109247347A CN 201811359515 A CN201811359515 A CN 201811359515A CN 109247347 A CN109247347 A CN 109247347A
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- volatile oil
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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]
- A01N65/36—Rutaceae [Rue family], e.g. lime, orange, lemon, corktree or pricklyash
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K17/00—Soil-conditioning materials or soil-stabilising materials
- C09K17/14—Soil-conditioning materials or soil-stabilising materials containing organic compounds only
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- C—CHEMISTRY; METALLURGY
- C09—DYES; PAINTS; POLISHES; NATURAL RESINS; ADHESIVES; COMPOSITIONS NOT OTHERWISE PROVIDED FOR; APPLICATIONS OF MATERIALS NOT OTHERWISE PROVIDED FOR
- C09K—MATERIALS FOR MISCELLANEOUS APPLICATIONS, NOT PROVIDED FOR ELSEWHERE
- C09K2101/00—Agricultural use
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Abstract
The invention discloses a kind of new application of Fructus Citri Sarcodactylis volatile oil, i.e. application of the Fructus Citri Sarcodactylis volatile oil in preparation prevention and treatment Chinese medicine or the chemical fertilizer or pesticide of crop root rot；The experimental results showed that Fructus Citri Sarcodactylis volatile oil can inhibit 3 kinds of main pathogenic bacteria Fusarium oxysporums for causing notoginseng root rot to occur, Fusarium solani and damage the mould mycelia growth of column spore；And Fructus Citri Sarcodactylis volatile oil is compounded to have bacterial strain with hymexazol and sums it up and act synergistically；Fructus Citri Sarcodactylis volatile oil is added in continuous cropping soil, the disease incidence and disease index of notoginseng root rot can be reduced；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 new application of Fructus Citri Sarcodactylis volatile oil more particularly to Fructus Citri Sarcodactylis volatile oils to Chinese medicine or crop root rot
Preventive and therapeutic effect belongs to biological pesticide technical field.
Background technique
Radix Notoginseng (Panax notoginseng(Burk.) F.H.Chen) also known as pseudo-ginseng, it is that Araliaceae Panax is perennial
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 at me
State southwest, wherein in Wenshan Prefecture of Yunnan Province away from modern existing more than 400 years cultivation histories.Roots of Panax Notoginseng, leaf, is spent enter stem
Medicine, scope of medication is very extensive, and social required quantity is being continuously increased, and with increasing sharply for notoginseng planting area, disease is asked
Topic 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 the factors such as characteristic, weather conditions and plantation tradition, thus inevitably there is a large amount of continuous cropping, cause yield and product
Matter 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.
Fingered citron (Citrus medica L.var.sarcodactylisIt Swingle) is rutaceae, evergreen shrubs
Or dungarunga, it is typically used as Chinese medicine, or because its fruit shape is peculiar, and as ornamental plant.
Any report of the Fructus Citri Sarcodactylis volatile oil for crop pest control is had no at present.
Summary of the invention
The purpose of the present invention is to provide a kind of new applications of Fructus Citri Sarcodactylis volatile oil, are the applications in field of biological pesticide,
Application i.e. in preparation prevention and treatment Chinese medicine or the chemical fertilizer or pesticide of crop root rot.
The purposes of Fructus Citri Sarcodactylis volatile oil of the present invention is that Fructus Citri Sarcodactylis volatile oil is used as to preparation prevention and treatment Chinese medicine or root of the crop
The preparation of maize ear rot prevents and treats the application in the bacteria preparation that causes a disease in preparation that is, using Fructus Citri Sarcodactylis volatile oil as active constituent.
The ingredient (or effective component) of the present invention for preventing and treating the bacteria preparation that causes a disease is Fructus Citri Sarcodactylis volatile oil, and one kind can also be added
Or acceptable auxiliary material on plurality of Chinese and crop disease control preparation, or played with other active components compounding and cooperate with suppression
Bacterium effect.
Fructus Citri Sarcodactylis volatile oil can be prepared into the pesticide of various dosage forms or the chemical fertilizer of various forms by prior art device.
Determine the inhibiting effect that Fructus Citri Sarcodactylis volatile oil grows spore germination and mycelia in the present invention, Fructus Citri Sarcodactylis volatile oil with
The MIC value of hymexazol is the same order of magnitude, and fungistatic effect is suitable.Fructus Citri Sarcodactylis volatile oil is compounded with chemical pesticide hymexazol,
It was found that it all has adduction and synergistic effect to the fungistatic effect of multiple bacterial strains；Increased by Fructus Citri Sarcodactylis volatile oil and chemical pesticide collaboration
Effect achievees the purpose that reduce chemical pesticide application.
Fructus Citri Sarcodactylis volatile oil is extracted by the conventional steam way of distillation in the present invention, and anhydrous sodium sulfate drying and volatilizing is oily and makes
, the preparation method is simple, and required reagent cost is low, is conducive to mass production.
Fructus Citri Sarcodactylis 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 Citri Sarcodactylis volatile oil is Extracts from Plant Recourses, the complicated multiplicity of effective component is right
Pathogen is the mechanism of action of multicomponent multiple target point, so that target organisms are not likely to produce to certain or certain anti-medicine of several ingredients generations
Property, to obtain lasting control efficiency；Compared with chemical synthetic pesticide, the effective component of Fructus Citri Sarcodactylis volatile oil is volatile, easy drop
Solution, it is antibacterial without sterilizing, therefore be not easy to cause damages to other beneficial microbes, improve soil micro-ecosystem structure, promotes to plant
Object and microorganism coevolution, preserve the ecological environment, and improve the quality of Chinese medicine or crop.
The present invention opens the new method for reducing chemical pesticide application, facilitates scientific worker to the volatile oil mechanism of action
Research, have broader prospect in agricultural production；The Fructus Citri Sarcodactylis volatile oil of the present invention for reducing the generation of Radix Notoginseng disease has
The characteristics of putting into low, easy to operate, high-efficient, volatile, less toxic, low-residual；The big back of organic agriculture is greatly developed in country
Under scape, fingered citron hair oil is the more satisfactory chemical agent substitute product for overcoming Radix Notoginseng disease, is had to the sustainable development of agricultural
Significance.
Detailed description of the invention
Fig. 1 be Fructus Citri Sarcodactylis volatile oil of the present invention and other compounds to Fusarium oxysporum (Fusarium oxysporum), it is rotten
Skin sickle-like bacteria (Fusarium solani) and damage column spore it is mould (Cylindrocarpon destructans) bacterium colony growth shadow
It rings result (colony diameter)；Wherein a is γ-terpinene；B is (R)-4-isopropenyl-1-methyl-1-cyclohexene；C is γ-terpinene+(R)-4-isopropenyl-1-methyl-1-cyclohexene；D waves for fingered citron
Hair oil；E is negative control (aqueous solution of the DMSO containing volumetric concentration 1% and the Tween 80 of volumetric concentration 0.1%)；F is powder azoles
Alcohol；G is hymexazol；
Fig. 2 be Fructus Citri Sarcodactylis volatile oil of the present invention and other compounds to Fusarium oxysporum (Fusarium oxysporum), Beancurd sheet
Sickle-like bacteria (Fusarium solani) and damage column spore it is mould (Cylindrocarpon destructans) bacterium colony growth influence
As a result (bacteriostasis rate)；
Fig. 3 is the disease incidence and disease index of Radix Notoginseng；A is the Radix Notoginseng of continuous cropping soil plantation；B is to volatilize containing 0.3mg/g fingered citron
The Radix Notoginseng of the continuous cropping soil plantation of oil.
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 Citri Sarcodactylis volatile oil
(1) add 8 times of water of powder quality in the Finger Citron that 300 g are crushed, impregnate 2 h, by the way of heating distillation, utilize
Volatile oil in steam extraction flos caryophylli, extraction time 6h, condensation collect and slip out liquid；
(2) volatile oil extracted is dry with anhydrous sodium sulfate, removes institute's water content, puts brown sample bottle into, seals ,-
20 DEG C save for use.
Embodiment 2: the activation of pathogenic bacteria
(1) separate Panax notoginseng root root rot pathogenic bacteria, Purification be Fusarium oxysporum (Fusarium oxysporum), it is rotten
Skin sickle-like bacteria (Fusarium solani) and damage column spore it is mould (Cylindrocarpon destructans), strain is inoculated with
It is activated on to PDA culture medium, after inoculation 4 times, takes eugonic bacterial strain spare；
(2) potato dextrose agar (PDA): 200 g of potato, 20 g of glucose, 20 g of agar, distilled water 1000
mL。
The chemical constituent of embodiment 3:GC-MS measurement Fructus Citri Sarcodactylis volatile oil
(1) GC condition: instrument model: Agilengt Technologies 7890B-5977B；Chromatographic column: 30 m of HP-5MS
×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；
A: retention time；B: relative peak area ± relative deviation；C: retention index.
Embodiment 4: the influence that measurement Fructus Citri Sarcodactylis volatile oil grows 3 kinds of pathogenic bacteria mycelia in embodiment 2
(1) under aseptic technique, 15mL PDA culture medium is poured into each culture dish, after solidification to be cooled, takes 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 5 mm of diameter along colony edge and takes fungus block, are inoculated with
In culture medium central；
(2) 4 sterile Oxford cups are equidistantly placed at fungi block 25mm；
(3) DMSO containing volumetric concentration 1% and the volumetric concentration 0.1% of the Fructus Citri Sarcodactylis volatile oil by above-described embodiment 1 after dry
The aqueous dissolution of Tween 80, it is 50mg/mL Fructus Citri Sarcodactylis volatile oil that concentration, which is made,；
(4) the 50mg/mL Fructus Citri Sarcodactylis volatile oil liquid of 200 μ L steps (3) is added (in advance with 0.22 μm of organic system in each Oxford cup
Filter filtration sterilization), 28 DEG C of constant temperature incubations in microbiological incubator, the DMSO containing volumetric concentration 1% of volatile oil is not added
Aqueous solution and hymexazol with the Tween 80 of volumetric concentration 0.1% are control, and 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 flos caryophylli volatile oil has apparent suppression to the mycelia growth of 3 kinds of pathogenic bacteria from Fig. 1,2
Production is used, and suitable with the effect of chemical agent hymexazol, wherein the fungistatic effect mould to Fusarium solani, breaking-up column spore is better than
Hymexazol.
Embodiment 5: Fructus Citri Sarcodactylis volatile oil MIC measurement
(1) 1 Fructus Citri Sarcodactylis volatile oil of embodiment is contained to the aqueous solution of the DMSO of volumetric concentration 1% and the Tween 80 of volumetric concentration 0.1%
Dissolution, the multiple concentration gradient liquid for being 75-0.15mg/mL with doubling dilution configuration concentration range, positive drug hymexazol, list
Body compound (R)-4-isopropenyl-1-methyl-1-cyclohexene and γ-terpinene are configured to the concentration ladder that concentration range is 2.5-0.005 mg/mL with same method
Degree；
(2) pathogen that 7d is grown in embodiment 2 rinses culture medium with 20 mL1/4PDA fluid nutrient mediums (not adding agar),
It is 1 × 10 that spore concentration, which is made,4The spore suspension of a/mL；
(3) system in the 50 μ L volatile oil and 150 μ L steps (2) of 0.22 μm of filtering with microporous membrane degerming is added in 96 orifice plates
At spore suspension；
(4) negative control is the aqueous solution and 150 that 50 μ L contain the DMSO of volumetric concentration 1% and the Tween 80 of volumetric concentration 0.1%
Manufactured spore suspension in μ L step (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 595nm
(Thermo 1510) measures the absorbance in each hole, as a result see the table below；
Note: with a line, different letters indicate that different disposal has significant difference (P < 0.05).
Embodiment 6: Fructus Citri Sarcodactylis volatile oil and hymexazol interaction measure
(1) MIC according to embodiment 5 obtained Fructus Citri Sarcodactylis volatile oil and hymexazol, being configured to maximum concentration is 8 times of MIC, is then adopted
8 dilution gradients are configured with doubling dilution；
(2) by the combination addition of the hymexazol (25 μ L) of the Fructus Citri Sarcodactylis 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) the MIC measurement of drug combination: corresponding drug concentration is grown as Fructus Citri Sarcodactylis volatile oil and hymexazol using no fungi hole and is joined
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；
(7) FIC of FICI=volatile oil FIC+ hymexazol, wherein FICI≤0.5 is synergistic effect, and 0.5 < FICI≤1 is adduction
Effect, 1<FICI≤4 are unrelated effect, and FICI>4 are antagonism, as a result be see the table below；
。
Embodiment 7: Inhibition test of the Fructus Citri Sarcodactylis volatile oil to the external disease incidence of Radix Notoginseng
Fructus Citri Sarcodactylis volatile oil is admixed in continuous cropping soil (continuous plantation 3 years or more notoginseng soil) in the ratio of 0.3mg/g and is placed
One week, after being sufficiently mixed the two uniformly, by healthy Radix Notoginseng seedling direct transplantation into the continuous cropping soil for being mixed with Fructus Citri Sarcodactylis volatile oil；Together
When healthy Radix Notoginseng seedling planted as compareing using continuous plantation 3 years or more notoginseng soil for not adding Fructus Citri Sarcodactylis volatile oil；
80 plants of Radix Notoginseng of every group of plantation, after plantation 30 days, carry out grade classification according to the withered degree of plant: 0 is without morbidity disease
Shape；1 wilts for partial blade；2 show the symptom wilted for all blades；3 symptoms wilted for stem performance；4 for blade and
Stem all shows wilting symptom.
Disease incidence and disease index calculate as follows:
Disease incidence=(morbidity strain number/total strain number) × 100%；
Disease index=100 × ∑ (morbidity strain number × respective level at different levels)/(investigation total strain number × superlative degree typical value)；
Experimental result see the table below and Fig. 3, as can be seen from the figure will volatilize oil and is added in continuous cropping soil, can reduce Radix Notoginseng root-rot
The disease incidence and disease index of disease, and since the effumability of volatile oil, fragrance keep away dirty feature, volatile oil safety of the present invention,
Effectively；
。
Claims (1)
1. application of the Fructus Citri Sarcodactylis volatile oil in preparation prevention and treatment Chinese medicine or the chemical fertilizer or pesticide of crop root rot.
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| CN201811359515.5A CN109247347B (en) | 2018-11-15 | 2018-11-15 | Application of fingered citron volatile oil |
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|---|---|---|---|
| CN201811359515.5A CN109247347B (en) | 2018-11-15 | 2018-11-15 | Application of fingered citron volatile oil |
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| CN109247347B CN109247347B (en) | 2021-04-23 |
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2007329367A1 (en) * | 2006-12-06 | 2008-06-12 | Benjamin Bogomolny | Composition for improving plant growth and yield and method for using same |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| AU2007329367A1 (en) * | 2006-12-06 | 2008-06-12 | Benjamin Bogomolny | Composition for improving plant growth and yield and method for using same |
Non-Patent Citations (2)
| Title |
|---|
| M.C. MANGANYI ET AL: ""Antimicrobial activities of selected essential oils against Fusarium oxysporum isolates and their biofilms"", 《SOUTH AFRICAN JOURNAL OF BOTANY》 * |
| V. C. PAWAR Æ V. S. THAKER: ""Evaluation of the anti-Fusarium oxysporum f. sp cicer and anti-Alternaria porri effects of some essential oils"", 《WORLD J MICROBIOL BIOTECHNOL 》 * |
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