CN102177893B - Bactericide - Google Patents
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- CN102177893B CN102177893B CN 201110068242 CN201110068242A CN102177893B CN 102177893 B CN102177893 B CN 102177893B CN 201110068242 CN201110068242 CN 201110068242 CN 201110068242 A CN201110068242 A CN 201110068242A CN 102177893 B CN102177893 B CN 102177893B
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Abstract
The invention discloses a bactericide which comprises 2-methyl-methyl propionate, 2-methyl propioric acid, 1-vinyl-1-methyl-2,4-di(1-methyl vinyl)-cyclohexane, carypohyllene, 2, 6-dimethyl-6-(4-methyl-3-pentene)bicyclo[3.1.1]heptyl-2-alkenyl, 1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methyl ethylidene)azulene, 1,2,3,4,4a,5,6,8a-octahydro-4a, 8-dimethyl-2-(1-methyl ethyl)naphthaline, 3,3,7,11-tetramethyl tricyclo[6.3.0.0(2,4)]undecarbon-8-alkene and the like. The bactericide can be used for inhibiting or killing pathogenic microorganisms in the crop production process, mildew microorganisms of building materials and pathogenic microorganisms in human or animal living environments.
Description
The present invention is to be 2009.09.30 the applying date, and application number is dividing an application of 200910153513.5 " Muscodor platymiscium endogenetic fungus and uses thereof and bactericide ".
Technical field
The present invention relates to microbiology and bactericide field, specifically, the present invention relates to a kind of new Muscodor platymiscium endogenetic fungus and based on or be derived from its volatility mixture and uses thereof.
Background technology
Fungi is a huge family in the microorganism, and fungi is very close with human relation, and fungi can be in order to make important product in the industrial or agricultural such as medicine, food, chemical industry.Famous penicillin and cephalosporin are that fungi brings one of most precious medicine of the world of medicine up to now, also are present most important antibiotic, are widely used in the treatment of bacterial infective diseases, and have saved thousands upon thousands mankind's life.
Plant endogenesis epiphyte (endophytic fungi) is the important monoid of fungi, estimates that according to Dreyfuss and Chapel the endogenetic fungus sum reaches 1,000,000 kinds.Existing a few is used to medicine, agronomy production or commercial Application aspect now, but endophytic fungus resources of plants is not also fully excavated far away.Endogenetic fungus has diversified Ecological Functions, and it is relevant with its diversity that the Ecological Functions of endogenetic fungus is likely, and the diversity of endogenetic fungus means its Chemical Diversity.Endogenetic fungus can produce various secondary metabolites, and they have important application potential in human lives, production, can produce the antitumoral compounds such as taxol such as endogenetic fungus; The novel anti-tumor actives such as oreganic acid; Noval chemical compound cryptocin, the cryptocandin etc. of the disease funguses such as anti-candida albicans, alpha fungus and trichophyton purpureatum; Generation has periconicins, the rhizoctonic acid etc. of the pathogenetic bacterias such as anti-helicobacter pylori, staphylococcus aureus, kerekou pneumonia Salmonella and salmonella typhimurium.
Now found that multiple different microorganism has the bioactivator that produces the control plant disease, comprises insecticide, bactericide and plant growth regulator etc.The at present research for the pathogenic microorganism of controlling pathogenic microorganism, construction material mildew micro-organism and the mankind or animal living enviroment in the crops production process also has many, and important progress is arranged, but mainly still rely on chemical synthesis process to produce bactericide or insecticide.Use the chemosynthesis medicament to bring a series of serious consequence: 1) high residue: because the chemosynthesis medicament comprises that its multiple raw material have carcinogenicity and toxicity, enter and be not easy eliminating in the animal and plant body, the environment degradable ability causes persticide residue high; 2) destroy the ecological balance: biological long-term evolution and adaptation, the various biological relative populations of occurring in nature reach dynamic equilibrium, use chemical agent to cause the natural enemy quantity of pest to greatly reduce; 3) drug-fast generation: the pathogen of occurring in nature and the gene of insects have very strong sudden change ability, and Irrational Use of Drugs, the medication of the region between the heart and the diaphragm order impel drug-fast rapid generation.Because above-mentioned all shortcomings, the use of chemosynthesis medicament is also more and more restricted, develop new, mechanism of action uniqueness and people and animals and the harmless biologic product of ecotope more and more are taken seriously, and also becomes the important directions of modern industry development.
Many kinds of fungies can produce some light-concentration volatile gases, and some escaping gas also has antibiotic, desinsection or viricidal function, such as Dennis﹠amp; Webster etc. find that the escaping gas substance of wood mould (Trichoderma) generation suppresses the growth of Rhizoctonia solani Kuhn (Rhizoctonia solani), Fusarium oxysporum (Fusarium oxysporum), Pythium ultimum (Pythium ultimum) etc.Strobel etc. are separated to from tropical woody plant can produce volatility compounding substances (volatile compounds, VOCs) four kinds of Muscodor belong to endogenetic fungus, and they are respectively cinnamon Ceylon cinnamon (Cinnamomum zeylanicum) endogenetic fungus Muscodor albus, fern leaf silvery birch (Grevillea pteridifolia) endogenetic fungus Muscodor roseus, torrid zone bejuco (Paullinia paullinioides) endogenetic fungus Muscodor vitigentis, Ananas plant Ananas ananassoides endogenetic fungus Muscodor crispans.The VOCs that Muscodor albus, Muscodor roseus, Muscodor vitigentis and Muscodor crispans produce has inhibitory or killing effect to fungi, bacterium and virus etc., have important using value in control or the pathogenic microorganism killed in pathogenic microorganism, construction material mildew micro-organism and the mankind or the animal living enviroment in the crops production process, they and also carried out relevant commercial development.
Utilize the application study of mycetogenetic VOCs at present also rare, Strobel etc. are about achievement in research and the Application and Development of living fungi inside the Pass the Muscodor symbolic animal of the birth year, it is the important channel of seeking new compound, newtype drug or newtype drug intermediate that proof Muscodor belongs to mycetogenetic VOCs, also is the valuable source of development of new biologic product.Utilize mycetogenetic VOCs to control the pathogenic microorganism of pathogenic microorganism, construction material mildew micro-organism and the mankind or animal living enviroment in the crops production process, have the non-target organism nonhazardous, free from environmental pollution, the not broken ring ecological balance, the advantage that can not induce harmful microorganism, virus and insect etc. to develop immunity to drugs.
Summary of the invention
The technical problem to be solved in the present invention provides a kind of bactericide, and it can suppress or kill the pathogenic microorganism in pathogenic microorganism, construction material mildew micro-organism and the mankind or the animal living enviroment in the crops production process.
In order to solve the problems of the technologies described above, the invention provides a kind of bactericide, this bactericide comprises 0.65%～10.2%2-methyl-methyl propionate, 28%～78%2-methylpropanoic acid, 0.6%～2.5%1-vinyl-1-methyl-2.4-two (1-methyl ethylene)-cyclohexane, 0.10%～0.4% caryophyllene, 0.06%～1%2,6-dimethyl-6-(4-methyl-3-amylene) two ring [3.1.1] hept-2-ene"s, 0.8%～3.5%1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methyl ethylidene) Azulene, 5.8%～11%1,2,3,4,4a, 5,6,8a-octahydro-4a, 8-dimethyl-2-(1-Methylethyl) naphthalene, 0.05%～0.2% 3,3,7,11-tetramethyl three ring [6.3.0.0 (2,4)] 11 carbon-8-alkene, 0.3%～2.8%a-phellandrene, 4%～45% β-phellandrene, 0.1%～0.6% trans 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.08%～0.4% cis 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.25%～4% trans 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.03%～0.28% cis 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol; All the other are solvent; Above percentage is percent by volume.
A kind of improvement as bactericide of the present invention: this bactericide comprises 9.5%～10.2%2-methyl-methyl propionate, 60%～61%2-methylpropanoic acid, 2.0%～2.1%1-vinyl-1-methyl-2.4-two (1-methyl ethylene)-cyclohexane, 0.36%～0.4% caryophyllene, 0.9%～1%2,6-dimethyl-6-(4-methyl-3-amylene) two ring [3.1.1] hept-2-ene"s, 3.3%～3.4%1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methyl ethylidene) Azulene, 6.9%～7.0%1,2,3,4,4a, 5,6,8a-octahydro-4a, 8-dimethyl-2-(1-Methylethyl) naphthalene, 0.13%～0.17% 3,3,7,11-tetramethyl three ring [6.3.0.0 (2,4)] 11 carbon-8-alkene, 0.3%～0.35%a-phellandrene, 5.8%～5.95% β-phellandrene, 0.2%～0.25% trans 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.08%～0.12% cis 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.25%～0.32% trans 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.05%～0.07% cis 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol; All the other are solvent.
Another kind as bactericide of the present invention improves: this bactericide comprises 0.68%～0.72%2-methyl-methyl propionate, 28%～28.5%2-methylpropanoic acid, 2%～2.5%1-vinyl-1-methyl-2.4-two (1-methyl ethylene)-cyclohexane, 0.10%～0.14% caryophyllene, 0.12%～0.18%2,6-dimethyl-6-(4-methyl-3-amylene) two ring [3.1.1] hept-2-ene"s, 2%～2.5%1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methyl ethylidene) Azulene, 10.26%～10.76%1,2,3,4,4a, 5,6,8a-octahydro-4a, 8-dimethyl-2-(1-Methylethyl) naphthalene, 0.14%～0.18% 3,3,7,11-tetramethyl three ring [6.3.0.0 (2,4)] 11 carbon-8-alkene, 2.38%～2.8%a-phellandrene, 43.5%～45% β-phellandrene, 0.5%～0.54% trans 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.34%～0.38% cis 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 3.85%～4% trans 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.22%～0.26% cis 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol; All the other are solvent.
Another improvement as bactericide of the present invention: this bactericide comprises 7.8%～8.2%2-methyl-methyl propionate, 76.5%～78%2-methylpropanoic acid, 0.63%～0.7%1-vinyl-1-methyl-2.4-two (1-methyl ethylene)-cyclohexane, 0.10%～0.12% caryophyllene, 0.06%～0.08%2,6-dimethyl-6-(4-methyl-3-amylene) two ring [3.1.1] hept-2-ene"s, 0.8%～0.9%1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methyl ethylidene) Azulene, 5.8%～6.2%1,2,3,4,4a, 5,6,8a-octahydro-4a, 8-dimethyl-2-(1-Methylethyl) naphthalene, 0.05%～0.07% 3,3,7,11-tetramethyl three ring [6.3.0.0 (2,4)] 11 carbon-8-alkene, 0.3%～0.4%a-phellandrene, 4%～4.5% β-phellandrene, 0.1%～0.15% trans 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.08%～0.12% cis 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.3%～0.35% trans 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.03%～0.05% cis 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol; All the other are solvent.
Solvent is conventional solvent, such as being: acetone, cyclohexane, methyl alcohol etc.
It can suppress or kill the pathogenic microorganism in pathogenic microorganism, construction material mildew micro-organism and the mankind or the animal living enviroment in the crops production process.
Plant endogenesis epiphyte Muscodor sp. bacterial strain, the preservation name is called: ZJLQ023, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center, preservation date: on January 8th, 2009, preserving number: CGMCC 2862.
Hereinafter to be referred as endogenetic fungus Muscodorsp. bacterial strain ZJLQ023.
The volatility compounding substances (VOCs) that endogenetic fungus Muscodor sp. bacterial strain ZJLQ023 produces is effective bactericide, to Rhizoctonia solani Kuhn, Pythium ultimum, Fusarium oxysporum, alternaria tenuis (Alternaria alternata), aspergillus (Aspergillus sp), Botrytis cinerea (Botrytis cinerea), Botrytis fabae (Botrytis fabae), circular thorn dish spore (Colletotrichum orbiculare), the bryonia Asia is every spore shell (Didymella bryoniae), Fulvia fulva (Fulvia fulva), Paecilomyces lilacinus (Paecilomyces lilacinus), Penicillium digitatum (Penicillum digitatum), persimmon dish stey (Pestalotia diospyri), piricularia oryzae (Pyricularia oryzae), Sclerotium rolfsii (Sclerotium rolfsii), sclerotinite (Sclerotinia sclerotiorum), verticillium dahliae (Verticillium dahliae), saccharomyces cerevisiae (Saccharomyces cerevisiae), the former thread fungies of Various Diseases such as Escherichia coli (Escherichia coli) and leaf bacillus (Phyllobacterium sp.), saccharomycete and Gram-negative bacteria and gram-positive bacteria all have inhibition or killing action.
Endogenetic fungus Muscodor sp. bacterial strain ZJLQ023 has that to produce VOCs speed fast, the characteristics that lasting effect is long, and the cell wall of can degrade Pythium ultimum and piricularia oryzae, and this VOCs has unique joyful fragrance of people that makes.
Endogenetic fungus Muscodorsp. bacterial strain ZJLQ023 of the present invention can utilize barley meal, fine rice bran, millet powder and sucrose etc. to be carbon source, can utilize peptone, dusty yeast, yeast extract, wheat bran, beancake powder and Dried Corn Steep Liquor Powder etc. to be nitrogenous source, can grow at cereal or wheat class, potato dextrose agar, oat agar glucose or brewer's wort agar glucose, and produce the VOCs with bacteriostatic activity.
The active carrier preparation of endogenetic fungus Muscodor sp. bacterial strain ZJLQ023 of the present invention has the business development purposes, the VOCs of ZJLQ023 culture and generation thereof can for the treatment of or prevent poisonous mould in construction material and the building, can be used for the fresh-keeping mildew-resistant of fruit and food, can be used for preventing or killing harmful mould of grain and feed, processing, flowers or the fruit that can be used for agricultural production soil are dredged the control of disease.
VOCs of the present invention can separate and obtains from being grown in novel endogenetic fungus Muscodorsp. bacterial strain ZJLQ023 culture on cereal or wheat class, potato dextrose agar, oat agar glucose or the brewer's wort agar glucose, the VOCs main active component except:
(a) 2-methyl-methyl propionate (Propanoic acid, 2-methyl-, methyl ester);
(b) 2 Methylpropionic acid (Propanoic acid, 2-methyl-);
(c) 1-vinyl-1-methyl-2.4-two (1-methyl ethylene)-cyclohexane (Cyclohexane, 1-ethenyl-1-methyl-2,4-bis (1-methylethenyl)-, [1S-(1.alpha., 2.beta., 4.beta.)]-);
(d) caryophyllene (Caryophyllene);
(e) 2,6-dimethyl-6-(4-methyl-3-amylene) two ring [3.1.1] hept-2-ene"s (Bicyclo[3.1.1] hept-2-ene, 2,6-dimethyl-6-(4-methyl-3-pentenyl)-);
(f) 1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methyl ethylidene) Azulene (Azulene, 1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methylethenyl)-, [1S-(1.alpha., 4.alpha., 7.alpha.)]-);
(g) 1,2,3,4,4a, 5,6,8a-octahydro-4a, 8-dimethyl-2-(1-Methylethyl) naphthalene (Naphthalene, 1,2,3,4,4a, 5,6,8a-octahydro-4a, 8-dimethyl-2-(1-methylethenyl)-, [2R-(2.alpha., 4a.alpha., 8a.beta.)]-);
(h) 3,3,7,11-tetramethyl three ring [6.3.0.0 (2,4)] 11 carbon-8-alkene (Tricyclo[6.3.0.0 (2,4)] undec-8-ene, 3,3,7,11-tetramethyl-), importantly contain:
(i) a-phellandrene (alpha.-Phellandrene);
(j) β-phellandrene (beta.-Phellandrene);
(k) trans 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, (2-Cyclohexen-1-ol, 1-methyl-4-(1-methylethyl)-, trans-)
(l) cis 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol (2-Cyclohexen-1-ol, 1-methyl-4-(1-methylethyl)-, cis-)
(m) trans 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol (2-Cyclohexen-1-ol, 3-methyl-6-(1-methylethyl)-, trans-)
(n) cis 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol (2-Cyclohexen-1-ol, 3-methyl-6-(1-methylethyl)-, cis-) class isoreactivity material.
Above-mentioned VOCs can be used as fumigant as the inhibition of the pathogenic microorganism in pathogenic microorganism, construction material mildew micro-organism and the mankind or the animal living enviroment in the crops production process or kills, and has the great potential of therefrom developing novel, efficient, nontoxic natural bactericidal agent.
The VOCs that produces in Muscodor sp.ZJLQ023 live body preparation of the present invention or the endogenetic fungus Muscodor sp ZJLQ023 incubation and comprise one or more phellandrenes or the purposes of the synthesizing fungicide of 2-cyclohexene-1-01 derivatives.They are applicable to industry, agricultural, aquaculture and service industry etc.For example they can process or prevent poisonous mould in construction material and the building; Can be used for the fresh-keeping mildew-resistant of fruit and food; Can be used for preventing or killing harmful mould of grain and feed; Processing, flowers or the fruit that can be used for agricultural production soil are dredged the control of disease.
Description of drawings
Fig. 1 is that Muscodor sp bacterial strain ZJLQ023 volatility mixture VOCs prevents and treats the rotten Test Drawing of the rear fruit of gathering; A, Muscodor sp bacterial strain ZJLQ023 culture; B, blank.
Embodiment
The VOCs that embodiment 1, endogenetic fungus Muscodor sp. bacterial strain ZJLQ023 produce analyzes
The VOCs that endogenetic fungus Muscodor sp. bacterial strain ZJLQ023 produces adopts SPE/gas chromatography/mass spectrometry technology (Solid phase microextraction/Gas chromatograph/Mass spetra, SPME/GC/MS) analysis.With Muscodor sp. bacterial strain ZJLQ023 inoculation on the PDA medium, cultivate 3-5d for 25 ℃, cutting the bacterium cake is transferred on PDA or the MEA medium, with sealed membrane the disposable plastic culture dish is sealed, cultivate certain hour (3d, 5d, 10d) in 25 ℃ of biochemical cultivation cases, bore an aperture with the fine needle head from the culture dish edge.SPME Stainless Steel protection needle tubing is inserted the Sample Room of GC/MS; release extracting head; extracting head is exposed in the GC vaporizer; 20 minutes kinds of 240 ℃ of activation; afterwards extracting head is retracted and protect needle tubing; insert immediately in the off-the-shelf culture dish to be measured, release extracting head, extracting head is exposed in the culture dish; can not run into mycelia; extraction was retracted extracting head immediately and is protected needle tubing after 45 minutes under the room temperature, inserted immediately the Sample Room of GC/MS; release extracting head; extracting head exposes in the GC vaporizer, and chromatographic determination is carried out in 240 ℃ of desorbs 30 seconds.Analysis operation condition and equipment are as follows:
Instrument, reagent and GC/MS condition: solid-phase micro-extracting device (U.S. Supelco company), extracting head coating are 50/30 μ M DVB/CAR/PDMS; Gas chromatograph-mass spectrograph (Agilent 6890N GC/5975B inert XL MS), quartz capillary column HP-5MS (30m * 250 μ m * 0.25 μ m); Data retrieval is used the NIST05 database.
Chromatographic condition: carrier gas is helium, post flow 1.0mLmin
-1Temperature of vaporization chamber: 240 ℃, 30 ℃ of chromatographic column initial temperatures keep 3min, with 5 ℃ of min
-1Heating rate rises to 220 ℃.
The mass spectrum condition: the EI ionization source, ionization voltage 70eV, 230 ℃ of ion source temperatures, 150 ℃ of quadrupole rod temperature, 280 ℃ of interface temperature, sweep limits: 20-450amu does not shunt.
Analyze by GC/MS, obtain the total ion current figure of each composition, determine the chemical composition of each component according to standard diagram and related documents, and calculate the relative percentage composition of each component by the peak area normalization method, Measurement results sees Table 1:
Table 1 endogenetic fungus Muscodor sp. bacterial strain ZJLQ023 and Muscodor albus cultivate the VOCs that 5d produces
Main antibacterial active compounds by:
(a) 2-methyl-methyl propionate (Propanoic acid, 2-methyl-, methyl ester);
(b) 2 Methylpropionic acid (Propanoic acid, 2-methyl-);
(c) 1-vinyl-1-methyl-2.4-two (1-methyl ethylene)-cyclohexane (Cyclohexane, 1-ethenyl-1-methyl-2,4-bis (1-methylethenyl)-, [1S-(1.alpha., 2.beta., 4.beta.)]-);
(d) caryophyllene (Caryophyllene);
(e) 2,6-dimethyl-6-(4-methyl-3-amylene) two ring [3.1.1] hept-2-ene"s (Bicyclo[3.1.1] hept-2-ene, 2,6-dimethyl-6-(4-methyl-3-pentenyl)-);
(f) 1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methyl ethylidene) Azulene (Azulene, 1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methylethenyl)-, [1S-(1.alpha., 4.alpha., 7.alpha.)]-);
(g) 1,2,3,4,4a, 5,6,8a-octahydro-4a, 8-dimethyl-2-(1-Methylethyl) naphthalene (Naphthalene, 1,2,3,4,4a, 5,6,8a-octahydro-4a, 8-dimethyl-2-(1-methylethenyl)-, [2R-(2.alpha., 4a.alpha., 8a.beta.)]-);
(h) 3,3,7,11-tetramethyl three ring [6.3.0.0 (2,4)] 11 carbon-8-alkene (Tricyclo[6.3.0.0 (2,4)] undec-8-ene, 3,3,7,11-tetramethyl-), importantly contain:
(i) a-phellandrene (alpha.-Phellandrene);
(j) β-phellandrene (beta.-Phellandrene);
(k) trans 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, (2-Cyclohexen-1-ol, 1-methyl-4-(1-methylethyl)-, trans-)
(l) cis 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol (2-Cyclohexen-1-ol, 1-methyl-4-(1-methylethyl)-, cis-)
(m) trans 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol (2-Cyclohexen-1-ol, 3-methyl-6-(1-methylethyl)-, trans-)
(n) cis 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol (2-Cyclohexen-1-ol, 3-methyl-6-(1-methylethyl)-, cis-) class isoreactivity material.
The VOCs control that embodiment 2, Muscodor sp. bacterial strain ZJ LQ023 produce is gathered, and rear fruit is rotten to be tested
For the preventive and therapeutic effect that the VOCs that tests Muscodor sp. bacterial strain ZJLQ023 generation rots to adopting rear fruit, embodiment is as follows: will control with the active carrier preparation Muscodor sp. bacterial strain ZJLQ023 that is grown on the PDA medium or prepares the rotten problem of the rear fruit of gathering.Particularly, the picking rear fruit without wound and insect pest (Xinjiang bergamot pear or honey peach) of gathering, place fruit conserving case (salable), scalpel is standardized wound gently, access brown rot germ (Monilinia fructicola) at wound, and apply moistening filter paper at wound and be used for moisturizing, the active carrier preparation Muscodor sp. bacterial strain ZJLQ023 that is grown on the PDA medium or prepares is positioned in the fruit conserving case, be used for suppressing the growth of pathogen, seal, put into 25 ℃ climatic cabinate, keep humidity 85%.Every processing 3 repeats, and not add Muscodor sp. bacterial strain ZJLQ023 culture as to blank, takes out fruit behind the 3-5d, investigation incidence and Taking Pictures recording.The result shows that the VOCs that Muscodor sp. bacterial strain ZJLQ023 culture produces can control the rotten problem (Fig. 1) of the rear fruit of gathering preferably.
The VOCs that embodiment 3, Muscodor sp. bacterial strain ZJLQ023 produce is to the preventive and therapeutic effect of kind of Seedling Diseases
Measure the VOCs of Muscodor sp. bacterial strain ZJLQ023 generation to the preventive and therapeutic effect of kind of Seedling Diseases with the radish seedling growth method.With grind Hou De Bite stone pack into diameter 50mm, the height 400mm test tube in, the height of Zhuan Ru Bite stone is about 10cm, 121 ℃ of moist heat sterilization 20min.Access pathogen Pythium ultimum bacterium cake (diameter 8mm) 5-6 piece ， Yong Bite stone covers in each test tube, and 10 seeds (ten No. three radish of short leaf, Guangdong Province's breeding is introduced service company and provided) are provided on it, and Zai Yong Bite stone covers.Hang up Muscodor sp. bacterial strain ZJLQ023 active carrier preparation (pack with gauze the outside) in the test group test tube, only to connect pathogen as blank, test tube seals with ventilative cork.The immigration climatic cabinate (25 ℃, illumination: dark=12h: cultivate 12h), regularly add up the incidence of germination rate and observation seedling.
The VOCs that table 2Muscodor sp. bacterial strain ZJLQ023 produces is to the preventive and therapeutic effect of kind of Seedling Diseases
The VOCs that Muscodor Sp bacterial strain ZJLQ023 active carrier preparation produces can suppress Pythium ultimum effectively on the impact of radish seed sprouting, growth of seedling.
The artificial synthetic mixture bactericide of embodiment 4, the separable VOCs that produces from Muscodor sp. bacterial strain ZJLQ023 culture
The applicant finds that (1) Muscodor sp. bacterial strain ZJLQ023 culture produces the basic all the components of VOCs; (2) Muscodorsp. bacterial strain ZJLQ023 culture produces the combination of part composition among the VOCs, or a kind of synthetic VOCs of composition has the sterilization idiocratic of Muscodor sp. bacterial strain ZJLQ023 among the generation VOCs of (3) Muscodorsp bacterial strain ZJLQ023 culture.
In order to test the relative biologically active of classes of compounds and classes of compounds optimum concentration and ratio in entire mixture, entire mixture is the detection mixture of the 100 μ L in every 50mL spatial domain on the culture in the standard culture flat board.For example, β-phellandrene accounts for identifies 5.88% of volatile matter mixture, with the test of 5.88 μ L/50mL (0.1176 μ L/mL) spatial domain β-phellandrene is tested, and other compound also is with identical standard procedure.
Following percentage is percent by volume.
With 10.18%2-methyl-methyl propionate, the 60.42%2-methylpropanoic acid, 2.04%1-vinyl-1-methyl-2.4-two (1-methyl ethylene)-cyclohexane, 0.38% caryophyllene, 0.95%2,6-dimethyl-6-(4-methyl-3-amylene) two ring [3.1.1] hept-2-ene"s, 2,6-dimethyl-6-(4-methyl-3-pentenyl), 3.35%1,2,3,4,5,6,7,8-octahydro-Isosorbide-5-Nitrae-dimethyl-7-(1-methyl ethylidene) Azulene, 6.93%1,2,3,4,4a, 5,6,8a-octahydro-4a, 8-dimethyl-2-(1-Methylethyl) naphthalene, 0.15% 3,3,7,11-tetramethyl, three ring [6.3.0.0 (2,4)] 11 carbon-8-alkene, 15.6% acetone is mixed with organic mixture (being called composition 1).
If the composition 1 of basic, normal, high three levels mixes with a-phellandrene, β-phellandrene, trans 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, cis 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, trans 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol, cis 3-methyl-6-(1-the Methylethyl)-2-cyclohexene-1-alcohol of equal volume, artificial synthetic waving property of the Combination compound of test is to indicating the inhibition of pathogen, take Pythium ultimum as the cause of disease indicator bacteria, test result sees Table 3.
The artificial synthetic volatility mixture that composition 1 and other compound of three varying levels of table 3 is made into is to the inhibiting rate of Pythium ultimum
Can find out from test result, high, in, the composition 1 of low three levels and the a-phellandrene of equal volume, β-phellandrene, trans 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, cis 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, trans 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol, the escaping gas that cis 3-methyl-6-(1-Methylethyl)-the 2-cyclohexene-1-alcohol mixes is different to the inhibiting rate of Pythium ultimum, inhibiting rate when low-level when (0.851 μ L/mL) is minimum, and middle level (1.702 μ L/mL) and high level (3.404 μ L/mL) inhibiting rate all reach 100%.
Embodiment 5, artificial synthetic bactericide, it comprises the composition of percent by volume as described in Table 4, surplus is acetone.
5 embodiment of table 4, bactericide
Adopt the test of two compartment plate culture to cultivate the activity experiment of VOCs (as a comparison case) Suppressing phytopathogens of 5d generation with N1～N5 and the Muscodor albus that is equal to table 1.The cause of disease indicator bacteria: Botrytis cinerea, Rhizoctonia solani Kuhn, alternaria tenuis, neat bulbil, Pythium ultimum, Fusarium oxysporum cucumber specialized form, circular thorn dish spore, verticillium dahliae, sclerotinite, persimmon dish stey, piricularia oryzae, Penicillium digitatum, Botrytis fabae, Fulvia fulva, bryonia Asia are every spore shell, aspergillus, Paecilomyces lilacinus.The cause of disease indicator bacteria is connected on the PDA medium activates, cut mycelia piece (diameter 5mm) (aspergillus is inoculated with spore suspension) and be inoculated in PDA medium (90mm has every a culture dish) side, to add respectively culture dish opposite side (every kind of amount of mixture is respectively as 100 μ L) take 6 kinds of mixtures of various compound ratios among the VOCs (as a comparison case) that cultivates the 5d generation according to N1～N5 and the Muscodoralbus that is equal to table 1, with the sealing of parafilm film, 25 ℃ of biochemical cultivation cases are cultivated.When treating that the pathogen bacterium colony grows to fast completely ware in the blank ware, measure the artificial volatility compounding substances VOCs that synthesizes to the growth inhibition of cause of disease indicator bacteria, calculate inhibiting rate, the result is as shown in table 5.
Table 5
Claims (4)
1. bactericide, it is characterized in that: described bactericide comprises 0.65%～10.2%2-methyl-methyl propionate, 28%～78%2-methylpropanoic acid, 0.6%～2.5%1-vinyl-1-methyl-2,4-two (1-methyl ethylene)-thiacyclohexane, 0.10%～0.4% caryophyllene, 0.06%～1% 2,6-dimethyl-6-(4-methyl-3-amylene) two ring [3.1.1] hept-2-ene"s, 0.8%～3.5%1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methyl ethylidene) Azulene, 5.8% ~ 11%1,2,3,4,4a, 5,6,8a-octahydro-4a, 8-dimethyl-2-(1-Methylethyl) naphthalene, 0.05%～0.2% 3,3,7,11-tetramethyl three ring [6.3.0.0 (2,4)] 11 carbon-8-alkene, 0.3%～2.8% α-phellandrene, 4%～45% β-phellandrene, 0.1%～0.6% trans 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.08%～0.4% cis 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.25%～4% trans 3-methyl-6-(1-Methylethyl)-pure and mild 0.03%～0.28% cis 3-methyl-6-(1-Methylethyl of 2-cyclohexene-1-)-2-cyclohexene-1-alcohol; All the other are solvent; Above percentage is percent by volume.
2. a kind of bactericide according to claim 1, it is characterized in that: described bactericide comprises 9.5%～10.2%2-methyl-methyl propionate, 60%～61%2-methylpropanoic acid, 2.0%～2.1%1-vinyl-1-methyl-2,4-two (1-methyl ethylene)-thiacyclohexane, 0.36%～0.4% caryophyllene, 0.9%～1%2,6--dimethyl-6-(4-methyl-3-amylene) two ring [3.1.1] hept-2-ene"s, 3.3%～3.4%1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methyl ethylidene) Azulene, 6.9% ~ 7.0%1,2,3,4,4a, 5,6,8a-octahydro-4a, 8-dimethyl-2-(1-Methylethyl) naphthalene, 0.13%～0.17% 3,3,7,11-tetramethyl three ring [6.3.0.0 (2,4)] 11 carbon-8-alkene, 0.3%～0.35% α-phellandrene, 5.8%～5.95% β-phellandrene, 0.2%～0.25% trans 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.08%～0.12% cis 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.25%～0.32% trans 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.05%～0.07% cis 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol; All the other are solvent.
3. a kind of bactericide according to claim 1, it is characterized in that: described bactericide comprises 0.68%～0.72%2-methyl-methyl propionate, 28%～28.5%2-methylpropanoic acid, 2%～2.5%1-vinyl-1-methyl-2,4-two (1-methyl ethylene)-thiacyclohexane, 0.10%～0.14% caryophyllene, 0.12%～0.18% 2,6-dimethyl-6-(4-methyl-3-amylene) two ring [3.1.1] hept-2-ene"s, 2%～2.5%1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methyl ethylidene) Azulene, 10.26% ~ 10.76%1,2,3,4,4a, 5,6,8a-octahydro-4a, 8-dimethyl-2-(1-Methylethyl) naphthalene, 0.14%～0.18% 3,3,7,11-tetramethyl three ring [6.3.0.0 (2,4)] 11 carbon-8-alkene, 2.38%～2.8% α-phellandrene, 43.5%～45% β-phellandrene, 0.5%～0.54% trans 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.34%～0.38% cis 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 3.85%～4% trans 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.22%～0.26% cis 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol; All the other are solvent.
4. a kind of bactericide according to claim 1, it is characterized in that: described bactericide comprises 7.8%～8.2%2-methyl-methyl propionate, 76.5%～78%2-methylpropanoic acid, 0.63%～0.7%1-vinyl-1-methyl-2,4-two (1-methyl ethylene)-thiacyclohexane, 0.10%～0.12% caryophyllene, 0.06%～0.08%2,6-dimethyl-6-(4-methyl-3-amylene) two ring [3.1.1] hept-2-ene"s, 0.8%～0.9%1,2,3,4,5,6,7,8-octahydro-1,4-dimethyl-7-(1-methyl ethylidene) Azulene, 5.8% ~ 6.2%1,2,3,4,4a, 5,6,8a-octahydro-4a, 8-dimethyl-2-(1-Methylethyl) naphthalene, 0.05%～0.07% 3,3,7,11-tetramethyl three ring [6.3.0.0 (2,4)] 11 carbon-8-alkene, 0.3%～0.4% α-phellandrene, 4%～4.5% β-phellandrene, 0.1%～0.15% trans 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.08%～0.12% cis 1-methyl-4-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.3%～0.35% trans 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol, 0.03%～0.05% cis 3-methyl-6-(1-Methylethyl)-2-cyclohexene-1-alcohol; All the other are solvent.
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