CN1337460A - Microorganism (Tr13) and preparation method thereof - Google Patents
Microorganism (Tr13) and preparation method thereof Download PDFInfo
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- CN1337460A CN1337460A CN 00109996 CN00109996A CN1337460A CN 1337460 A CN1337460 A CN 1337460A CN 00109996 CN00109996 CN 00109996 CN 00109996 A CN00109996 A CN 00109996A CN 1337460 A CN1337460 A CN 1337460A
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- trichoderma
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- 244000005700 microbiome Species 0.000 title claims abstract description 12
- 238000002360 preparation method Methods 0.000 title abstract description 5
- 241000196324 Embryophyta Species 0.000 claims abstract description 19
- 239000002689 soil Substances 0.000 claims abstract description 15
- 241000223259 Trichoderma Species 0.000 claims abstract description 14
- 208000031888 Mycoses Diseases 0.000 claims abstract description 11
- 235000015097 nutrients Nutrition 0.000 claims abstract description 8
- 238000004321 preservation Methods 0.000 claims abstract description 4
- 241000908011 Trichoderma aureoviride Species 0.000 claims abstract description 3
- 239000007788 liquid Substances 0.000 claims description 12
- 238000012360 testing method Methods 0.000 claims description 9
- 241000894006 Bacteria Species 0.000 claims description 6
- 244000061456 Solanum tuberosum Species 0.000 claims description 6
- 235000002595 Solanum tuberosum Nutrition 0.000 claims description 6
- 238000012258 culturing Methods 0.000 claims description 6
- 238000000855 fermentation Methods 0.000 claims description 6
- 230000004151 fermentation Effects 0.000 claims description 6
- 238000005286 illumination Methods 0.000 claims description 6
- 238000000034 method Methods 0.000 claims description 6
- 238000004519 manufacturing process Methods 0.000 claims description 5
- 229920001817 Agar Polymers 0.000 claims description 3
- 229930182555 Penicillin Natural products 0.000 claims description 3
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 claims description 3
- 239000008272 agar Substances 0.000 claims description 3
- 230000000845 anti-microbial effect Effects 0.000 claims description 3
- 230000003115 biocidal effect Effects 0.000 claims description 3
- 229940041514 candida albicans extract Drugs 0.000 claims description 3
- 230000003203 everyday effect Effects 0.000 claims description 3
- 229940049954 penicillin Drugs 0.000 claims description 3
- LKPLKUMXSAEKID-UHFFFAOYSA-N pentachloronitrobenzene Chemical compound [O-][N+](=O)C1=C(Cl)C(Cl)=C(Cl)C(Cl)=C1Cl LKPLKUMXSAEKID-UHFFFAOYSA-N 0.000 claims description 3
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 claims description 3
- 229960001225 rifampicin Drugs 0.000 claims description 3
- 239000002002 slurry Substances 0.000 claims description 3
- 239000007787 solid Substances 0.000 claims description 3
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 238000004659 sterilization and disinfection Methods 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 238000009736 wetting Methods 0.000 claims description 3
- 239000002023 wood Substances 0.000 claims description 3
- 239000012138 yeast extract Substances 0.000 claims description 3
- 230000003042 antagnostic effect Effects 0.000 abstract description 5
- 238000011161 development Methods 0.000 abstract description 5
- 239000000126 substance Substances 0.000 abstract description 4
- 238000006243 chemical reaction Methods 0.000 abstract description 3
- 108010059892 Cellulase Proteins 0.000 abstract description 2
- 229920002101 Chitin Polymers 0.000 abstract description 2
- 102000012286 Chitinases Human genes 0.000 abstract description 2
- 108010022172 Chitinases Proteins 0.000 abstract description 2
- 208000035240 Disease Resistance Diseases 0.000 abstract description 2
- 229940106157 cellulase Drugs 0.000 abstract description 2
- 239000001913 cellulose Substances 0.000 abstract description 2
- 229920002678 cellulose Polymers 0.000 abstract description 2
- 238000003912 environmental pollution Methods 0.000 abstract description 2
- 230000035558 fertility Effects 0.000 abstract description 2
- 239000003337 fertilizer Substances 0.000 abstract description 2
- 239000002207 metabolite Substances 0.000 abstract description 2
- 239000000575 pesticide Substances 0.000 abstract description 2
- 230000008635 plant growth Effects 0.000 abstract description 2
- 238000011160 research Methods 0.000 abstract description 2
- 229930192334 Auxin Natural products 0.000 abstract 1
- 239000002363 auxin Substances 0.000 abstract 1
- 230000009286 beneficial effect Effects 0.000 abstract 1
- SEOVTRFCIGRIMH-UHFFFAOYSA-N indole-3-acetic acid Chemical compound C1=CC=C2C(CC(=O)O)=CNC2=C1 SEOVTRFCIGRIMH-UHFFFAOYSA-N 0.000 abstract 1
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- 244000061176 Nicotiana tabacum Species 0.000 description 2
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- 230000001580 bacterial effect Effects 0.000 description 2
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- 241000283690 Bos taurus Species 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
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- 239000003375 plant hormone Substances 0.000 description 1
- 244000144977 poultry Species 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention relates to a microorganism and a preparation method thereof. It is characterized in that the Trichoderma reeoviride is Trichoderma Tr13(Trichoderma aureoviride) for biological control of plant fungal diseases, and the preservation registration number is CGMCC NO. 0440. By adopting the antagonistic microbe of the invention to carry out biological control, on one hand, the produced metabolites such as cellulose and chitin of plant residues in soil can be decomposed by using cellulase, chitinase and the like, so that the content of nutrient components in the soil is increased, the conversion of organic matters in the soil is promoted, and the produced auxin is beneficial to the development of plant root systems, the growth of plants and the induction of disease resistance reaction of the plants. But also can improve the soil fertility, reduce the environmental pollution caused by chemical fertilizers and pesticides, has no harm to people and livestock, short research period and low investment, is easy for industrialized commercialized development and creates conditions for sustainable development of agriculture.
Description
The present invention relates to microbial technology field, specifically a kind of microorganism and preparation method thereof.
Because present most of plants comprise tobacco, flowers, vegetables and lack the disease-resistant variety to fungal disease that the control of fungal diseases of plants is still at present based on chemical pesticide control.Yet continuous use causes pesticide residue and then human body generation harm and the drug-fast generation of pathogenic bacteria is caused that people pay close attention to greatly.Take measures to limit use, registration and the registration updating of some huge malicious agricultural chemicals in the world gradually.USDA by rules according to the document; " when facing the disease and pest problem, whenever possible, the method that biological control is always preferentially selected.Also there is not at present a kind of effective biological control method for fungal diseases of plants such as tobacco, flowers, vegetables.
The objective of the invention is at there not being effective antagonistic microbe to be used to prevent and treat fungal diseases of plants at present, and provide a kind of from soil screening and culturing go out specific antagonistic microbe and prevent and treat fungal diseases of plants, the present invention also provides this microorganism preparation method.
The biological control of Plant diseases utilizes antagonistic microbe to prevent and treat the disease of plant exactly.For separation screening effectively to inhibiting biocontrol strain can be arranged the germ of above-mentioned various fungal diseases of plants, the present invention has taked special soil mass trapping, by making bait with germ itself and selecting substratum, face-off culture method to screen Trichoderma Tr13 (Trichoderma aureoviride), in specified depositary institution of Patent Office of the People's Republic of China (Chinese microorganism strain management committee common micro-organisms center) preservation, its preservation registration number is CGMCC NO.0440 to this bacterial classification.
The Tr13 bacterial strain has following character:
Cultural characteristic:
PDA and on the MA substratum 25 ℃, observed in 3-10 days, bacterium colony is crawled to grow and is formed a large amount of white aerial hyphaes soon, in early days, suede is cotton-shaped, conidium clump interfascicular is given birth to wherein, presents the spot film circular permutation, is initially canescence, sap green when aging, the aerial hyphae of this moment covers whole bacterium colony surface pistac like spider web; The dirty yellow in the MA substratum bacterium colony back side, PDA substratum back side light color.
The microscopy form:
The mycelia branch is complicated, separation is arranged, smooth colourless, the mycelium of wall contains oil droplet sample material, diameter 2-6 μ m, and chlamydospore is rare;
Conidiophore: the side shoot on the major branch is cluster growth clearly typically, mostly be living or alternate mode are formed secondary, three grades of branches, common second branch, form pinecone formula branch profile at last, usually the branch angle is slightly less than 90 degree, the bottle stalk 2-3 cluster of giving birth on branch, seat the male character types in Chinese operas, usu. referring tov the bearded character degree is big, and the direction of growth is tending towards the branch top;
The bottle stalk: shape is long, and base portion slightly is narrower than waist and thin, and from narrowing to form long-neck more than the waist, the bottle stalk waist that has is slightly crooked, presents asymmetric doleiform or " cattle horn shape ", 6-12 * 2-2.5 μ m, common 7-11 μ m * 2 μ m;
Conidium: wall is smooth, green, the obovate base portion, has obvious cross section, accidental avette 2.8-4 * 2-2.8 μ m.
The soil separation method of mentioned microorganism is:
Make bait and be close to the wetting soil surface of sterilized water with covering with the mycelial nylon mesh sheet of corresponding germ, illumination every day 12-15 hour, cultivate after 5-8 days down for 25-28 ℃, the net sheet is moved on on Martin's agar medium, in substratum, add penicillin, quintozene, Rifampin, make each culture dish (about 15-20ml substratum) contain above-mentioned antibiotic respectively and reach 50ppm, cultivate this Trichoderma.
The liquid fermentation production method of mentioned microorganism is characterized in that this production method carries out according to the following steps:
1) test tube substratum: adopt solid PDA potato culture, should the mould kind of wood be seeded on the test tube substratum, 27-29 cultivated 46-50 hour down;
2) enlarged culturing base: adopt liquid PDA potato culture, Trichoderma in the test tube is seeded in the Erlenmeyer flask, place on the shaking table 27-29 vibration illumination cultivation 72-76 hour as seed liquor;
3) liquid fermentation medium: adopt Li Chashi yeast extract paste nutrient solution, 121 sterilize when nutrient solution subject to sterilization was cooled to 27.5 ℃ in about 30 minutes, with the ratio access fermented liquid of the culture in the Erlenmeyer flask, cultivate for 27.5 ℃ and obtained corresponding antimicrobial bacterium slurry short of money in 48 hours in 5-7%.
The Trichoderma of above-mentioned gained is made microbial bactericide through certain technical process, is used for preventing and treating fungal diseases of plants.
Adopt antagonistic microbe of the present invention to carry out biological control, can utilize the meta-bolites of generation such as Mierocrystalline cellulose, the chitin that cellulase, chitinase etc. decompose plant residue in the soil on the one hand, increase the nutrient composition content in the soil, promote soil with organic matter to transform, the plant hormone that produces is of value to the growth of root system of plant again, promote plant-growth, the reaction of inducing plant disease resistance.They can produce antibacterial substance again in addition, suppress growth of pathogenic bacteria, and can in the soil of non-sterile, spread wide, thereby improving soil fertility, the environmental pollution that minimizing chemical fertilizer, agricultural chemicals cause is to the person poultry harmless, research cycle is short, drop into lowly, be easy to the industrialization commercialized development, for agricultural sustainable development creates conditions.
Embodiment:
Make bait with covering with the mycelial nylon mesh sheet of corresponding germ, be close to the wetting soil surface of sterilized water, illumination every day 12 hours, after cultivating the sky under 25 ℃, the net sheet is moved on on Martin's agar medium, in substratum, add penicillin, quintozene, Rifampin, make each culture dish (about 15ml substratum) contain above-mentioned each antibiotic and reach 50ppm respectively, as selecting substratum, screening and culturing goes out this Trichoderma with this.
The liquid fermentation production method of microorganism is:
1) test tube substratum: adopt solid PDA potato culture, should the mould kind of wood be seeded on the test tube substratum, cultivated 46-50 hour down for 27-29 ℃;
2) enlarged culturing base: adopt liquid PDA potato culture, Trichoderma in the test tube is seeded in the Erlenmeyer flask, place on the shaking table 27-29 ℃ of vibration illumination cultivation 72-76 hour as seed liquor;
3) liquid fermentation medium: adopt Li Chashi yeast extract paste nutrient solution, 121 ℃ of autoclavings are about 30 minutes, when nutrient solution subject to sterilization is cooled to 275 ℃, the culture in the Erlenmeyer flask is inserted fermented liquid in 6% ratio, cultivate for 27.5 ℃ and obtained corresponding antimicrobial bacterium slurry short of money in 48 hours.
Above-mentioned Trichoderma is made microbial bactericide through certain technical process, be used for preventing and treating fungal diseases of plants.
Claims (4)
1, a kind of microorganism is characterized in that it is the Trichoderma Tr13 (Trichoderma aureoviride) that is used for the fungal diseases of plants biological control, and its preservation registration number is CGMCC No.0440.
2, by the described method for culturing microbes of claim 1, it is characterized in that above-mentioned Trichoderma cultivates by the following method: make bait and be close to the wetting soil surface of sterilized water with covering with the mycelial net sheet of corresponding germ, illumination every day 12-15 hour, cultivate after 5-8 days down for 25-28 ℃, the net sheet is moved on on Martin's agar medium, in substratum, add penicillin, quintozene, Rifampin, make each culture dish contain above-mentioned antibiotic respectively and reach 50ppm, cultivate this Trichoderma.
3,, it is characterized in that this production method carries out according to the following steps by the liquid fermentation production method of claim 1 or 2 described microorganisms:
1) test tube substratum.Adopt solid PDA potato culture, be seeded on the test tube substratum wood is mould, cultivated 46-50 hour down for 27-29 ℃;
2) enlarged culturing base: adopt liquid PDA potato culture, Trichoderma in the test tube is seeded in the Erlenmeyer flask, place on the shaking table 27-29 ℃ of vibration illumination cultivation 72-76 hour as seed liquor;
3) liquid fermentation medium: adopt Li Chashi yeast extract paste nutrient solution, 121 ℃ of autoclavings are about 30 minutes, when nutrient solution subject to sterilization is cooled to 27.5 ℃,, cultivates for 27.5 ℃ and obtained corresponding antimicrobial bacterium slurry short of money in 48 hours the ratio access fermented liquid of the culture in the Erlenmeyer flask in 5-7%.
4, by the described application of microorganism of claim 1, it is characterized in that Trichoderma is used for preventing and treating fungal diseases of plants.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00109996 CN1337460A (en) | 2000-08-09 | 2000-08-09 | Microorganism (Tr13) and preparation method thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 00109996 CN1337460A (en) | 2000-08-09 | 2000-08-09 | Microorganism (Tr13) and preparation method thereof |
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| Publication Number | Publication Date |
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| CN1337460A true CN1337460A (en) | 2002-02-27 |
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| CN 00109996 Pending CN1337460A (en) | 2000-08-09 | 2000-08-09 | Microorganism (Tr13) and preparation method thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102071145A (en) * | 2010-09-09 | 2011-05-25 | 河南省农业科学院 | Trichoderma viride fungi and preparation and application of fungicide thereof |
| CN105733961A (en) * | 2016-04-18 | 2016-07-06 | 云南中烟工业有限责任公司 | Trichoderma aureoviride strain and method for preparing radix scrophulariae tobacco flavor from same |
-
2000
- 2000-08-09 CN CN 00109996 patent/CN1337460A/en active Pending
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102071145A (en) * | 2010-09-09 | 2011-05-25 | 河南省农业科学院 | Trichoderma viride fungi and preparation and application of fungicide thereof |
| CN102071145B (en) * | 2010-09-09 | 2012-09-05 | 河南省农业科学院 | Trichoderma viride fungi and preparation and application of fungicide thereof |
| CN105733961A (en) * | 2016-04-18 | 2016-07-06 | 云南中烟工业有限责任公司 | Trichoderma aureoviride strain and method for preparing radix scrophulariae tobacco flavor from same |
| CN105733961B (en) * | 2016-04-18 | 2019-05-17 | 云南中烟工业有限责任公司 | A kind of trichoderma aureoviride bacterial strain and the method for preparing radix scrophulariae tobacco aromaticss with it |
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