CN108371187B - Terpene volatile materials is inhibiting the application in strawberry spherical cavity bacterium - Google Patents
Terpene volatile materials is inhibiting the application in strawberry spherical cavity bacterium Download PDFInfo
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- CN108371187B CN108371187B CN201810039802.1A CN201810039802A CN108371187B CN 108371187 B CN108371187 B CN 108371187B CN 201810039802 A CN201810039802 A CN 201810039802A CN 108371187 B CN108371187 B CN 108371187B
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- Prior art keywords
- strawberry
- spherical cavity
- volatile materials
- linalool
- myrtenol
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- 235000016623 Fragaria vesca Nutrition 0.000 title claims abstract description 96
- 235000011363 Fragaria x ananassa Nutrition 0.000 title claims abstract description 96
- 239000000463 material Substances 0.000 title claims abstract description 77
- 241000894006 Bacteria Species 0.000 title claims abstract description 57
- 150000003505 terpenes Chemical class 0.000 title claims abstract description 49
- 235000007586 terpenes Nutrition 0.000 title claims abstract description 49
- 230000002401 inhibitory effect Effects 0.000 title claims abstract description 17
- 240000009088 Fragaria x ananassa Species 0.000 title 1
- 241000220223 Fragaria Species 0.000 claims abstract description 96
- CDOSHBSSFJOMGT-UHFFFAOYSA-N linalool Chemical compound CC(C)=CCCC(C)(O)C=C CDOSHBSSFJOMGT-UHFFFAOYSA-N 0.000 claims abstract description 74
- RXBQNMWIQKOSCS-UHFFFAOYSA-N (7,7-dimethyl-4-bicyclo[3.1.1]hept-3-enyl)methanol Chemical compound C1C2C(C)(C)C1CC=C2CO RXBQNMWIQKOSCS-UHFFFAOYSA-N 0.000 claims abstract description 68
- 239000001490 (3R)-3,7-dimethylocta-1,6-dien-3-ol Substances 0.000 claims abstract description 37
- CDOSHBSSFJOMGT-JTQLQIEISA-N (R)-linalool Natural products CC(C)=CCC[C@@](C)(O)C=C CDOSHBSSFJOMGT-JTQLQIEISA-N 0.000 claims abstract description 37
- 229930007744 linalool Natural products 0.000 claims abstract description 37
- RXBQNMWIQKOSCS-RKDXNWHRSA-N Myrtenol Natural products C1[C@H]2C(C)(C)[C@@H]1CC=C2CO RXBQNMWIQKOSCS-RKDXNWHRSA-N 0.000 claims abstract description 34
- 230000004763 spore germination Effects 0.000 claims abstract description 24
- 238000000034 method Methods 0.000 claims abstract description 11
- 230000002265 prevention Effects 0.000 claims abstract description 8
- 230000000813 microbial effect Effects 0.000 claims abstract description 7
- 238000002360 preparation method Methods 0.000 claims abstract description 5
- 239000002316 fumigant Substances 0.000 claims description 2
- 201000010099 disease Diseases 0.000 abstract description 8
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 8
- 230000000694 effects Effects 0.000 abstract description 6
- 238000012360 testing method Methods 0.000 abstract description 5
- 239000000575 pesticide Substances 0.000 abstract description 3
- 241000180222 Sitobion fragariae Species 0.000 description 24
- 244000052769 pathogen Species 0.000 description 17
- 230000001717 pathogenic effect Effects 0.000 description 17
- 239000000126 substance Substances 0.000 description 13
- 241000196324 Embryophyta Species 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 9
- XMGQYMWWDOXHJM-UHFFFAOYSA-N limonene Natural products CC(=C)C1CCC(C)=CC1 XMGQYMWWDOXHJM-UHFFFAOYSA-N 0.000 description 8
- 230000000844 anti-bacterial effect Effects 0.000 description 7
- 235000001510 limonene Nutrition 0.000 description 7
- 229940087305 limonene Drugs 0.000 description 7
- 230000006378 damage Effects 0.000 description 6
- 238000012545 processing Methods 0.000 description 6
- 230000000007 visual effect Effects 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 5
- 206010039509 Scab Diseases 0.000 description 4
- 241000270295 Serpentes Species 0.000 description 4
- 150000001875 compounds Chemical class 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 210000001700 mitochondrial membrane Anatomy 0.000 description 4
- UGTJLJZQQFGTJD-UHFFFAOYSA-N Carbonylcyanide-3-chlorophenylhydrazone Chemical compound ClC1=CC=CC(NN=C(C#N)C#N)=C1 UGTJLJZQQFGTJD-UHFFFAOYSA-N 0.000 description 3
- 210000004027 cell Anatomy 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 3
- 235000013399 edible fruits Nutrition 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 241000233866 Fungi Species 0.000 description 2
- 239000003899 bactericide agent Substances 0.000 description 2
- NEHNMFOYXAPHSD-UHFFFAOYSA-N citronellal Chemical compound O=CCC(C)CCC=C(C)C NEHNMFOYXAPHSD-UHFFFAOYSA-N 0.000 description 2
- 230000007423 decrease Effects 0.000 description 2
- 230000003247 decreasing effect Effects 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
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- 238000004519 manufacturing process Methods 0.000 description 2
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- 230000001629 suppression Effects 0.000 description 2
- JUUMAEXLYIMEOD-UHFFFAOYSA-N 2,6-dimethylhept-1-ene Chemical compound CC(C)CCCC(C)=C JUUMAEXLYIMEOD-UHFFFAOYSA-N 0.000 description 1
- 244000144730 Amygdalus persica Species 0.000 description 1
- 239000005739 Bordeaux mixture Substances 0.000 description 1
- ABTFODLFVYTBHS-YAIFJCAJSA-N C[C@H](CCC(=O)NC1=CC=CC=C1)[C@H]2CC[C@@H]3[C@@]2([C@H](C[C@H]4[C@H]3CC[C@H]5[C@@]4(CC[C@H](C5)O)C)O)C Chemical compound C[C@H](CCC(=O)NC1=CC=CC=C1)[C@H]2CC[C@@H]3[C@@]2([C@H](C[C@H]4[C@H]3CC[C@H]5[C@@]4(CC[C@H](C5)O)C)O)C ABTFODLFVYTBHS-YAIFJCAJSA-N 0.000 description 1
- 241001164374 Calyx Species 0.000 description 1
- TWFZGCMQGLPBSX-UHFFFAOYSA-N Carbendazim Natural products C1=CC=C2NC(NC(=O)OC)=NC2=C1 TWFZGCMQGLPBSX-UHFFFAOYSA-N 0.000 description 1
- WTEVQBCEXWBHNA-UHFFFAOYSA-N Citral Natural products CC(C)=CCCC(C)=CC=O WTEVQBCEXWBHNA-UHFFFAOYSA-N 0.000 description 1
- 235000007466 Corylus avellana Nutrition 0.000 description 1
- 240000003211 Corylus maxima Species 0.000 description 1
- 208000035240 Disease Resistance Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241001465805 Nymphalidae Species 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000006040 Prunus persica var persica Nutrition 0.000 description 1
- 241000589516 Pseudomonas Species 0.000 description 1
- 240000007994 Rhodomyrtus tomentosa Species 0.000 description 1
- 235000007234 Rhodomyrtus tomentosa Nutrition 0.000 description 1
- 244000061458 Solanum melongena Species 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 239000005842 Thiophanate-methyl Substances 0.000 description 1
- 240000000851 Vaccinium corymbosum Species 0.000 description 1
- 235000003095 Vaccinium corymbosum Nutrition 0.000 description 1
- 235000017537 Vaccinium myrtillus Nutrition 0.000 description 1
- 239000003905 agrochemical Substances 0.000 description 1
- 150000001336 alkenes Chemical class 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 230000004323 axial length Effects 0.000 description 1
- BESJRHHIPGWPTC-UHFFFAOYSA-N azane;copper Chemical compound N.[Cu] BESJRHHIPGWPTC-UHFFFAOYSA-N 0.000 description 1
- 230000003385 bacteriostatic effect Effects 0.000 description 1
- 235000021014 blueberries Nutrition 0.000 description 1
- 239000006013 carbendazim Substances 0.000 description 1
- JNPZQRQPIHJYNM-UHFFFAOYSA-N carbendazim Chemical compound C1=C[CH]C2=NC(NC(=O)OC)=NC2=C1 JNPZQRQPIHJYNM-UHFFFAOYSA-N 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004587 chromatography analysis Methods 0.000 description 1
- 229940043350 citral Drugs 0.000 description 1
- 229930003633 citronellal Natural products 0.000 description 1
- 235000000983 citronellal Nutrition 0.000 description 1
- 238000004140 cleaning Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 239000004205 dimethyl polysiloxane Substances 0.000 description 1
- 235000013870 dimethyl polysiloxane Nutrition 0.000 description 1
- 150000002085 enols Chemical class 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000004817 gas chromatography Methods 0.000 description 1
- 238000002290 gas chromatography-mass spectrometry Methods 0.000 description 1
- WTEVQBCEXWBHNA-JXMROGBWSA-N geranial Chemical compound CC(C)=CCC\C(C)=C\C=O WTEVQBCEXWBHNA-JXMROGBWSA-N 0.000 description 1
- PCHJSUWPFVWCPO-UHFFFAOYSA-N gold Chemical compound [Au] PCHJSUWPFVWCPO-UHFFFAOYSA-N 0.000 description 1
- 229910052737 gold Inorganic materials 0.000 description 1
- 239000010931 gold Substances 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 230000003993 interaction Effects 0.000 description 1
- 238000010813 internal standard method Methods 0.000 description 1
- 230000002147 killing effect Effects 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 230000017074 necrotic cell death Effects 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- CXQXSVUQTKDNFP-UHFFFAOYSA-N octamethyltrisiloxane Chemical compound C[Si](C)(C)O[Si](C)(C)O[Si](C)(C)C CXQXSVUQTKDNFP-UHFFFAOYSA-N 0.000 description 1
- 244000039328 opportunistic pathogen Species 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- 230000035479 physiological effects, processes and functions Effects 0.000 description 1
- 238000004987 plasma desorption mass spectroscopy Methods 0.000 description 1
- 229920000435 poly(dimethylsiloxane) Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000012552 review Methods 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 238000005070 sampling Methods 0.000 description 1
- 229920006395 saturated elastomer Polymers 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- QGHREAKMXXNCOA-UHFFFAOYSA-N thiophanate-methyl Chemical group COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC QGHREAKMXXNCOA-UHFFFAOYSA-N 0.000 description 1
- 239000000341 volatile oil Substances 0.000 description 1
Classifications
-
- 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
- A01N49/00—Biocides, pest repellants or attractants, or plant growth regulators, containing compounds containing the group, wherein m+n>=1, both X together may also mean —Y— or a direct carbon-to-carbon bond, and the carbon atoms marked with an asterisk are not part of any ring system other than that which may be formed by the atoms X, the carbon atoms in square brackets being part of any acyclic or cyclic structure, or the group, wherein A means a carbon atom or Y, n>=0, and not more than one of these carbon atoms being a member of the same ring system, e.g. juvenile insect hormones or mimics 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
- A01N31/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic oxygen or sulfur compounds
- A01N31/04—Oxygen or sulfur attached to an aliphatic side-chain of a carbocyclic ring system
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Pest Control & Pesticides (AREA)
- Plant Pathology (AREA)
- Health & Medical Sciences (AREA)
- Agronomy & Crop Science (AREA)
- Dentistry (AREA)
- General Health & Medical Sciences (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Environmental Sciences (AREA)
- Insects & Arthropods (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention discloses terpene volatile materials to inhibit the application in strawberry spherical cavity bacterium, which is linalool or myrtenol.Present invention firstly discloses in Strawberry Leaves linalool and myrtenol both terpene volatile materials be able to suppress strawberry spherical cavity bacterium, the especially spore germination of strawberry spherical cavity bacterium and mycelia growth, and the test by fumigating strawberry spherical cavity bacterium or Strawberry Leaves using the terpene volatile materials, demonstrate the effect of linalool and myrtenol in the spore germination and mycelia growth for effectively inhibiting strawberry spherical cavity bacterium, and the function in prevention and treatment white leaf spot of strawberry.The present invention prevents and treats the preparation of the microbial white leaf spot of strawberry of strawberry spherical cavity using linalool and myrtenol preparation, and carry out white leaf spot of strawberry disease control, control efficiency is obvious, it can effectively reduce the usage amount of pesticide, provide foundation to develop the biological control method of novel, efficient, green safe prevention and treatment white leaf spot of strawberry.
Description
Technical field
The invention belongs to technical field of agriculture science more particularly to terpene volatile materials in inhibiting strawberry spherical cavity bacterium
Using.
Background technique
Strawberry spherical cavity bacterium (M.fragariae) is the pathogen for causing white leaf spot of strawberry evil, the also known as small spherical cavity of strawberry snake eye
Bacterium belongs to Ascomycotina chamber Pseudomonas fungi, and Invisible element conidiophore grows thickly, shorter, filbert or colourless, branch or regardless of
Branch, base portion stroma are undeveloped.The oval tubular of conidium generally has 1~2 diaphragm, cell inclusion granule to spindle-like.
White leaf spot of strawberry is also known as strawberry Leucoplakia, leaf spot, and main harm blade, morbidity causes scab mostly in old leaf,
Also petiole, carpopodium, calyx, stolon and carpopodium are infected.There are many subcircular, purples to differ in size on blade in early stage
Red stigma, like snake eye shape.Later, central spot be transformed into it is greyish white to taupe and form blotch, there are also aubergines sometimes
Take turns line.When humidity is high, scab surface generates the mould layer of white powder, i.e. germ conidiophore and conidium.Later period is in scab
On can produce out many pores, i.e. germ ascostome.When serious generation, scab gathers on blade, and morbidity leaf is in the short time
Necrosis is dried-up.White leaf spot of strawberry is very big to the harm of strawberry fruit, seriously will cause and drops in production over a large area.
Currently, the controlling mode of white leaf spot of strawberry in addition to wheel for ploughing does, lowering of watertable, ventilated, cleaning sick seedling,
It extracts outside the traditional field cultivation means of sick leaf and fertilizition control etc., mainly or based on chemical control, such as Bordeaux mixture, generation
Gloomy MnZn green must protect suspending agent, network ammonia copper water, can kill, the agrochemicals such as Bravo, thiophanate methyl and carbendazim.But by
The problems such as pesticide residue caused by the long-time service of chemical bactericide, environmental pollution, food safety, pathogen drug resistance, is increasingly
Seriously, it so, researches and develops safe and efficient biological bactericide and is of great significance.
Plant itself can generate a variety of volatile materials, these volatile materials as airborne signal chemical, plant with
Very important effect is played in interaction between environment, plant and disease and plant and plant etc..Recently, research both at home and abroad
Show that internal volatile matter content can rise plant by pathogen infection or after mechanical damage occurs, terpene volatile materials
One type key components, confirm on evidence these substances play an important role in disease resistance of plant (Dudareva,
Klempien et al.2013)。
He Peiqing etc. (2005) review the volatile materials with antibacterial activity or component plant and corresponding cause of disease
Bacterium, but there is inhibitory activity for the active volatile ingredient in Strawberry Leaves and to strawberry spherical cavity bacterium (M.fragariae)
Volatile materials is not registered.Publication No. is that the application for a patent for invention document of CN104920359A discloses a kind of efficient prevention and control water
The plants essential oil of rice rice blast, principle active component are citral and citronellal.Publication No. is the invention of CN105123911A
Patent application discloses a kind of method for adopting rear gray mold using linalool prevention and treatment blueberry.
Summary of the invention
The purpose of the present invention is to provide terpene volatile materials to inhibit the application in strawberry spherical cavity bacterium, terpene volatilization
Property substance can effectively inhibit strawberry spherical cavity bacterium spore germination and mycelia growth, and then prevent and treat the microbial strawberry of strawberry spherical cavity
Snake eye disease, prepares pesticide control with terpene volatile materials existing in such Strawberry Leaves, can have efficient, safety and ring
The features such as border is friendly overcomes deficiency in the prior art.
Particular technique content is as follows:
The present invention provides terpene volatile materials to inhibit the application in strawberry spherical cavity bacterium, the terpene volatile materials
For linalool or myrtenol.
Further, terpene volatile materials is able to suppress the spore germination and mycelia growth of strawberry spherical cavity bacterium.
The present invention is tried using main two kinds of terpene volatile materials linalools in Strawberry Leaves and myrtenol
It tests, since linalool and myrtenol are volatile materials, so being all made of in bacteriostatic experiment and disease control closed
In the environment of carry out stifling mode to be handled, through experiments, it was found that, linalool and hill gooseberry's alkene under a certain concentration range
Alcohol can effectively inhibit the spore germination and mycelia growth of strawberry spherical cavity bacterium.
Wherein, in the spore germination for inhibiting strawberry spherical cavity bacterium, with the body of the periphery closed environment where strawberry spherical cavity bacterium
Product meter, the concentration of the linalool and myrtenol >=5 μ L/L.
In the mycelia growth for inhibiting strawberry spherical cavity bacterium, in terms of the volume of the periphery closed environment where strawberry spherical cavity bacterium,
The concentration of the linalool >=0.5 μ L/L, concentration >=5 μ L/L of the myrtenol.
When the concentration of linalool and myrtenol reaches 50 μ L/L, spore germination, spore germination rate can be effectively suppressed
Decline 80-99%；There is bactericidal effect higher than the terpene volatile materials of above-mentioned concentration, i.e. volatile materials is fumigated 16h and moved back
After volatile materials, spore continues culture 72h and still fails to sprout.The low dosage of above-mentioned terpene volatile materials is stifling also
The growth of pathogen mycelia after sprouting enough is significantly inhibited, wherein it is raw that mycelia can be significantly reduced when concentration is up to 5 μ L/L in linalool
Long by 12%, when 10 × higher concentration, mycelia growth is almost totally constrained.
In addition, film potential is to the energetic supersession and growth of mycelia and infects host and has very important effect, film potential
Largely reflect the physiological status of cell.In order to verify and probe into terpene volatile materials for the physiology of the pathogen
Influence, the film potential of different volatile materials treated mycelia be determined, film potential dyed by JC1 after by being total to
Focusing microscope is observed and carries out fluorescent quantitation.The result shows that it is corresponding with mycelia growth, as terpene volatile materials is dense
The film potential of the rising of degree, strawberry spherical cavity bacterium (M.fragariae) mycelia gradually declines, for concentration higher than 50 μ L/L processing
Afterwards, mycelia mitochondrial membrane potential is remarkably decreased.Terpene volatile materials can destroy the film potential of the pathogen, and then influence bacterium
The growth of body.
The present invention also provides terpene volatile materials in the system for preparing the prevention and treatment microbial white leaf spot of strawberry of strawberry spherical cavity
Application in agent, the terpene volatile materials are linalool or myrtenol；With the periphery closed environment where strawberry
The concentration of stereometer, the linalool and myrtenol is 5-150 μ L/L.
Specifically, the white leaf spot of strawberry is located on Strawberry Leaves.
The present invention also provides a kind of methods for preventing and treating the microbial white leaf spot of strawberry of strawberry spherical cavity, with terpene volatility object
Matter fumigates strawberry as fumigant；The terpene volatile materials is linalool or myrtenol；With the periphery where strawberry
The concentration of the stereometer of closed environment, the linalool and myrtenol is 5-150 μ L/L.
Preferably, stifling Strawberry Leaves.
Further, the concentration of the linalool and myrtenol is 50 μ L/L.Under the concentration, terpene volatility object
Matter shows best inhibitory effect and to blade without any damage.
Compared with prior art, the invention has the following advantages:
(1) present invention firstly discloses the linalools and myrtenol both terpene volatile materials in Strawberry Leaves
It is able to suppress strawberry spherical cavity bacterium, the especially spore germination of strawberry spherical cavity bacterium and mycelia growth, and by volatilizing using the terpene
Property substance fumigate the tests of strawberry spherical cavity bacterium or Strawberry Leaves, it was demonstrated that linalool and myrtenol are effectively inhibiting strawberry ball
Function in effect in the spore germination of chamber bacterium and mycelia growth, and prevention and treatment white leaf spot of strawberry.
(2) present invention prevents and treats the system of the microbial white leaf spot of strawberry of strawberry spherical cavity using linalool and myrtenol preparation
Agent, and carry out white leaf spot of strawberry disease control, control efficiency is obvious, can effectively reduce the usage amount of pesticide, for develop it is novel,
Efficiently, the biological control method of green safe prevention and treatment white leaf spot of strawberry provides foundation.
Detailed description of the invention
Fig. 1 is the form of strawberry spherical cavity bacterium (M.fragariae) under different growth conditions；
Wherein, A is not sprout spore；B is the mycelia that growth is inhibited by volatile materials；C is the bacterium after normal growth 16h
Silk；D is the mycelia sprouted by volatile materials inhibits but 72h sprouts after removal again.
Fig. 2 is the influence that D- limonene grows strawberry spherical cavity bacterium (M.fragariae) mycelia
Fig. 3 is the influence that linalool grows strawberry spherical cavity bacterium (M.fragariae) mycelia.
Fig. 4 is the influence that myrtenol grows strawberry spherical cavity bacterium (M.fragariae) mycelia.
Fig. 5 is influence of the terpene volatile materials to strawberry spherical cavity bacterium (M.fragariae) film potential；
Wherein, A is linalool；B is myrtenol.
Fig. 6 is the Strawberry Leaves lesion area that terpene volatile materials infects strawberry spherical cavity bacterium (M.fragariae)
It influences.
Specific embodiment
The measurement of main volatile compounds in 1 Strawberry Leaves of embodiment
1. experimental material
Fresh strawberry blade is derived from strawberry (Fragaria × ananassa, Albion), trade name " A Er ratio ".
2. experimental method
It is detected in Strawberry Leaves using Headspace solid phase microextractiom enrichment-internal standard method for gas chromatography method (GC-MS)
Main volatile materials and its content.
20g fresh strawberry blade is taken to mix simultaneously homogenate, precise 3g sample according to (1g:2.5mL) with saturated salt solution
It is placed in the head space bottle of 15mL, 1ul 300ppm is added, 2,6- dimethyl -6- heptene -2- alcohol are as internal standard；It is stirred at 40 DEG C
After balancing 15min, it is inserted into 50/30 μm of DVB/CAR/PDMS solid phase micro-extracting head (Supelco, Bellefonte, PA, USA) extraction
Take 30min.
After extraction, hand sampling, 250 DEG C of parsing 3min of injection port, using gassy system (6890N) series connection MS detection
Device (5975B) (Agilent, Santa Clara, CA, the U.S.) is detected.Chromatography column parameter: DB-5MS pillar (30m ×
0.25mm,0.25μm)；Column temperature program setting: 40 DEG C (retaining 3min), with 2 DEG C of min-164 DEG C are risen to, with 3 DEG C of min-1On
145 DEG C are raised to, then with 15 DEG C of min-1Rise to 250 DEG C (retaining 10min).
Each volatile component is identified by being compared with mass spectral database (NIST05), passes through interior scalar quantity.
3. interpretation of result
The main volatile compounds measured in strawberry detect three kinds of terpene substances as shown in table 1, are peach gold respectively
Ma's enol, linalool and a small amount of D- limonene, total content are about 5%.
Main volatile compounds and its content in 1 Strawberry Leaves of table
Antibacterial activity in view of terpene volatile materials is it has been reported that this invention is selected in such Strawberry Leaves and naturally deposited
Volatile materials alternately, meanwhile, be with reference to setting with the natural contents of these volatile materials in Strawberry Leaves
The tested concentration of subsequent subject volatile materials.
The inhibition that main volatile compounds grow strawberry spherical cavity bacterium spore germination and mycelia in 2 Strawberry Leaves of embodiment
1. experimental material
1) for trying pathogen
Strawberry spherical cavity bacterium (M.fragariae) picks up from the strawberry of nature leave piece snake eye disease, by laboratory point
From, purifying, identification after and save backup, spore used in present case is all within four generation of Strawberry Leaves culture medium culture.
2) reagent agent
(98%) Sigma-Aldrich, content are all larger than for D- limonene, linalool, myrtenol
2. experimental method
It is 5 × 10 by concentration4The 100 μ L of strawberry spherical cavity bacterium (M.fragariae) spore suspension of a/mL is uniformly coated on
In PDA culture medium (the every disk of 7mL culture medium, about 0.15cm is thick, and culture dish diameter is 90mm), stands 10min and spore is allowed to be attached to
Media surface；
The filter paper of a 90mm is placed in culture dish cover inside, then respectively drips the volatile materials of certain volume
It is added on filter paper, lid lid is returned into culture dish rapidly, and with the closed sealing of sealed membrane.
The volume for each volatile materials being added dropwise reaches 0.05,0.5,5,50,500 μ according to concentration in final culture dish
L/L is implemented, and for low concentration therein, each volatile materials needs to carry out 1:10,1 with water according to experiment before dropwise addition:
100、1:103、1:104After dilution, then corresponding volume is added dropwise；Each volatile materials grouping is tested, and every group of setting is only dripped
Add the control group of isometric water, the volatile materials volume of dropwise addition is 16 μ L-50 μ L, and is all volatilized in 8h.
9 visuals field are randomly selected after 16h under an optical microscope to be taken pictures (amplification factor 125), then using aobvious
Micro mirror capture software is measured and counts to each thallus length in the visual field；Thallus number in each visual field 6 with
On, take the average thallus length in each visual field parallel as one.
The measurement of Hyphal length: the thallus picture of 16h capture is using the matched image measurement software of microscope to thallus axis
It is measured to total length.
The judgement of spore germination: thallus elongation is that spore average axial length (about 41 μm) are considered as sprouting (> 82 twice or more
μm)；All spore numbers in the spore number/visual field sprouted in spore germination rate=visual field；
For the processing group that 16h does not sprout, filter paper is removed, and culture dish is opened, the 1h that divulges information on the super-clean bench allows phase
It answers volatile materials to remove, then seals culture 72h again, then observe spore germination situation；The volatilization if spore is sprouted again
Property substance under the concentration only have the function of inhibition spore germination, be considered as the volatile materials if spore is not sprouted yet at this
There is bactericidal effect under concentration.
Entire experiment is independent to be repeated twice.
3. interpretation of result
The aspect graph of the strawberry spherical cavity bacterium (M.fragariae) of different growth conditions, as shown in Figure 1.
A is not sprout spore；B is the mycelia that growth is inhibited by volatile materials；C is the mycelia after normal growth 16h；D
To sprout the mycelia that by volatile materials inhibition but 72h sprouts after removing again.
Influence of the volatile materials to strawberry spherical cavity bacterium (M.fragariae) spore germination in 2 Strawberry Leaves of table
* this volatile materials has bacteriostasis under the concentration: 72h spore is sprouted again after removal
This volatile materials has bactericidal effect under the Δ concentration: 72h spore is not sprouted yet after removal
1) Strawberry Leaves volatile materials D- limonene is raw to (M.fragariae) spore germination of strawberry spherical cavity bacterium and mycelia
Long influence, respectively as shown in table 2 and figure 2.
D- limonene concentration only reaches inhibited to spore germination when 500 μ L/L, and average germination rate is reduced to
0.60%, however the concentration of limonene that the concentration is far longer than in general environment and detects in blade, and smell is excessively strong
It is strong.When in environment D- limonene concentration 50 μ L/L and it is following when, to subject pathogen thalli growth do not play significant suppression
Production is used.
2) Strawberry Leaves volatile materials linalool grows (M.fragariae) spore germination of strawberry spherical cavity bacterium and mycelia
Influence, it is as shown in Table 2 and Fig. 3 respectively.
Linalool concentration has obvious inhibiting effect, and the higher suppression of concentration in 50 μ L/L and 500 μ L/L, to spore germination
Rate processed is higher, inhibits the sprouting of 80% and 100% spore respectively, and has the function of killing pathogen when 500 μ L/L；It is stifling
Immediately volatile materials is removed after processing, spore is also no longer sprouted.In addition, as shown in figure 3, when concentration is 5 μ L/L also to bacterium
Silk growth has inhibiting effect.
3) Strawberry Leaves volatile materials myrtenol is to (M.fragariae) spore germination of strawberry spherical cavity bacterium and mycelia
The influence of growth, respectively as shown in table 2 and Fig. 4.
Myrtenol just significantly inhibits the sprouting of pathogen when concentration reaches 50 μ L/L, but disease
The germination rate of opportunistic pathogen still has 20% or so, and concentration almost inhibits spore germination and show to sterilize in 500 μ L/L
Effect.Its in low concentration (5 μ L/L and following) cannot effectively inhibit the growth of thallus.
Influence of the 3 terpene volatile materials of embodiment to strawberry spherical cavity bacterium (M.fragariae) film potential
1. experimental material
1) for trying pathogen
Strawberry spherical cavity bacterium (M.fragariae), with case study on implementation 2.
2. experimental method
It 1) is 5 × 10 by concentration4Strawberry spherical cavity bacterium (M.fragariae) the spore suspension 100uL of a/mL is uniformly coated on
In PDA culture medium (the every disk of 20mL culture medium, about 0.15cm is thick, and culture dish diameter is 90mm), after culture 5 days, according to implementation case
Method in example 2 carries out terpene volatile materials suffocating treatment 16h.
After treatment, the picking mycelia from each group, using mitochondrial membrane potential detection kit (JC-1), (the green skies are raw
Object science and technology, Shanghai) mitochondrial membrane potential of mycelia is detected.
Concrete operations are carried out according to experimental procedure provided by kit, and fluorescence intensity passes through Laser Scanning Confocal Microscope SP8 (Lay
Card, Germany) it is detected.It is each to test each processing group in triplicate, and every group is done technology repetition three times.Final film potential knot
Fruit is with red fluorescence intensity (excitation wavelength 525nm, launch wavelength 590nm) and green fluorescence intensity (excitation wavelength 490nm, hair
The long 530nm of ejected wave) ratio be expression result as final film potential.10 μM of CCCP processing is used as positive control (CCCP
Meeting is handled so that mycelia film potential completely loses), water process is as blank control.
3. interpretation of result
The film potential of mycelia is remarkably decreased, and shows that terpene substances can destroy the film potential of the pathogen, and then influence bacterium
The growth of body.When linalool concentration reaches 500 μ L/L, the film potential of mycelia is almost lost, and ties with 10 μM of CCCP processing
Fruit is similar (such as Fig. 5).
4 terpene volatile materials of embodiment infects the inhibition of Strawberry Leaves to strawberry spherical cavity bacterium (M.fragariae)
1. experimental material
1) for trying Strawberry Leaves
Fresh A Er chooses the similar blade of growth conditions and carries out than the fresh strawberry blade of strawberry with embodiment 1
Test.
2) for trying pathogen
Strawberry spherical cavity bacterium (M.fragariae), with embodiment 2.
2. experimental method
The similar Strawberry Leaves of fresh, physiological status are placed in 90mm culture dish, and spread two layers of filter paper in blade it
Under, 1mL water is added dropwise to filter paper, to keep the humidity and intact physiological status of blade.
It is 1 × 10 by concentration5Strawberry spherical cavity bacterium (M.fragariae) spore suspension of a/mL be sprayed on face of blade and
Reverse side carries out artificial infection；After the completion of inoculation, the filter paper of a 90mm is placed in culture dish cover inside, then respectively by one
The volatile materials for determining volume is added dropwise on filter paper, lid lid is returned culture dish rapidly, and with the closed sealing of sealed membrane.Each terpene
The concentration of substance is respectively 0 (control), and 5,50,500 μ L/L are implemented.
Blade opens culture dish after handling 2h at room temperature, will with volatile materials filter paper remove, culture dish without
It is sealed again after collarium border lower open mouth 30min (sufficiently removal volatile materials).Strawberry is observed and recorded after being further cultured for 8 days at room temperature
The incidence of blade.
3. interpretation of result
The terpene volatile materials suffocating treatment of various concentration has the influence of highly significant to the disease incidence of white leaf spot of strawberry
(Fig. 6).When terpene substances concentration is that 5,50 μ L/L phases than compareing can inhibit the disease incidence of white leaf spot of strawberry, wherein 50 μ L/L
Effect is best, almost inhibits infecting for the pathogen.For being tested terpene volatile materials, optimal concentration is 50 μ
L/L。
However, the volatile materials of excessive concentrations is handled, damage is also brought along to Strawberry Leaves itself, when volatility terpene
Material concentration is 500 μ L/L, and treated, and maroon occurs in blade, shows that blade cell is damaged, more so as to cause blade
It is easy to be infected by pathogen, 8 days rear blade almost all are infected by strawberry spherical cavity bacterium (M.fragariae).
According to result above, when terpene substances linalool and myrtenol use concentration as 50 μ L/L, show most
Good inhibitory effect and to blade without any damage.
It mainly include linalool, myrtenol in conclusion the main terpene volatile materials in Strawberry Leaves,
Have the function of significantly inhibiting (M.fragariae) spore germination of strawberry spherical cavity bacterium and mycelia growth under certain ambient concentration,
And pathogen mycelia mitochondrial membrane potential can be made to decline, to influence the growth of fungi.
Under suitable concentration, terpene substances can effectively inhibit the intrusion of the pathogen, prevent and treat the hair of white leaf spot of strawberry
It is raw, and under preferred dosage: when terpene volatile materials is 50 μ L/L, to show best inhibitory effect and to blade without appointing
What is damaged.It is demonstrated experimentally that linalool and myrtenol have good preventive and therapeutic effect to white leaf spot of strawberry.
Claims (5)
1. terpene volatile materials is inhibiting the application in strawberry spherical cavity bacterium, which is characterized in that the terpene volatile materials is
Linalool or myrtenol；
It is described in terms of the volume of the periphery closed environment where strawberry spherical cavity bacterium in the spore germination for inhibiting strawberry spherical cavity bacterium
The concentration of linalool and myrtenol is 50-150 μ L/L；
It is described in terms of the volume of the periphery closed environment where strawberry spherical cavity bacterium in the mycelia growth for inhibiting strawberry spherical cavity bacterium
The concentration 5-150 μ L/L of linalool, the concentration 50-150 μ L/L of the myrtenol.
2. application of the terpene volatile materials in the preparation of the preparation prevention and treatment microbial white leaf spot of strawberry of strawberry spherical cavity, feature
It is, the terpene volatile materials is linalool or myrtenol；In terms of the volume of the periphery closed environment where strawberry,
The concentration of the linalool and myrtenol is 50-150 μ L/L.
3. the application as described in right 2, which is characterized in that the white leaf spot of strawberry is located on Strawberry Leaves.
4. a kind of method for preventing and treating the microbial white leaf spot of strawberry of strawberry spherical cavity, which is characterized in that with terpene volatile materials work
For fumigant, strawberry is fumigated；The terpene volatile materials is linalool or myrtenol；It is closed with the periphery where strawberry
The concentration of the stereometer of environment, the linalool and myrtenol is 50-150 μ L/L.
5. the method as described in right 4, which is characterized in that the concentration of the linalool and myrtenol is 50 μ L/L.
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| CN104041547A (en) * | 2014-05-29 | 2014-09-17 | 赵兰 | Natural bactericide for controlling strawberry leaf spot and preparation method thereof |
| CN105010429A (en) * | 2015-06-26 | 2015-11-04 | 安徽禾众农业科技有限公司 | Strawberry white leaf spot prevention and control bacteriostat, and preparation method and application thereof |
| CN105379748A (en) * | 2015-10-19 | 2016-03-09 | 中国农业科学院柑桔研究所 | Antimicrobial pesticide composition and application thereof |
| CN107343503A (en) * | 2017-07-24 | 2017-11-14 | 湖北工程学院 | The application of terpenoid and apply its product |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104041547A (en) * | 2014-05-29 | 2014-09-17 | 赵兰 | Natural bactericide for controlling strawberry leaf spot and preparation method thereof |
| CN105010429A (en) * | 2015-06-26 | 2015-11-04 | 安徽禾众农业科技有限公司 | Strawberry white leaf spot prevention and control bacteriostat, and preparation method and application thereof |
| CN105379748A (en) * | 2015-10-19 | 2016-03-09 | 中国农业科学院柑桔研究所 | Antimicrobial pesticide composition and application thereof |
| CN107343503A (en) * | 2017-07-24 | 2017-11-14 | 湖北工程学院 | The application of terpenoid and apply its product |
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