CN1337458A - Microorganism (T918) and preparation method thereof - Google Patents
Microorganism (T918) and preparation method thereof Download PDFInfo
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- CN1337458A CN1337458A CN 00109994 CN00109994A CN1337458A CN 1337458 A CN1337458 A CN 1337458A CN 00109994 CN00109994 CN 00109994 CN 00109994 A CN00109994 A CN 00109994A CN 1337458 A CN1337458 A CN 1337458A
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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 13
- 208000031888 Mycoses Diseases 0.000 claims abstract description 11
- 235000015097 nutrients Nutrition 0.000 claims abstract description 8
- 241001304120 Trichoderma pseudokoningii Species 0.000 claims abstract description 4
- 238000004321 preservation Methods 0.000 claims abstract description 4
- 239000007788 liquid Substances 0.000 claims description 12
- 238000012360 testing method Methods 0.000 claims description 9
- 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
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- 241000894006 Bacteria Species 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 4
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- 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
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 238000009736 wetting Methods 0.000 claims description 3
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- 239000012138 yeast extract Substances 0.000 claims description 3
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- 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
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- 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
- 241000223261 Trichoderma viride Species 0.000 abstract 1
- 239000002363 auxin Substances 0.000 abstract 1
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- 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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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention relates to a microorganism and a preparation method thereof. The Trichoderma viride strain is characterized by being Trichoderma T918(Trichoderma pseudokoningii) for biological control of plant fungal diseases, and the preservation registration number of the Trichoderma pseudokoningii strain is CGMCC NO. 0442. 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 T918 (Trichoderma pseudokoningii), 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.0442 to this bacterial classification.
The T918 bacterial strain has following character:
Cultural characteristic:
25 ℃ of cultivations in 5-10 days are observed on the MA substratum, bacterium colony diffusion growth, and aerial hyphae is few, forms closely knit gonimoblast bundle districts in a large number, and initial canescence becomes olive-green gradually, and the back side, aging back is significantly yellow; Cultivate to observe in 5 days for 25 ℃ on the PDA substratum, colony growth is fast, forms many and closely knit gonimoblast bundle district, and 3 open the back color transfers green to by white, become after aging olive colour, back side light color.
The microscopy form:
That the mycelium branch has is smooth every, wall, mycelium contains oil droplet sample material, and diameter 2-6 μ m is wide;
Chlamydospore exists in a large number, and a life or top are born in the substrate mycelium, common fusion phenomenon, sphere, pyriform, ellipse, 5.6-15 μ m * 5-15 μ m;
Conidiophore: branch is simpler, major branch prolongs growth, upright, the common right angle of secondary branch stretches out, (dendritic) that crooked growth is also arranged like rainbow, secondary branch and single living bottle stalk with unequal-interval and are born on the major branch, mostly be Dan Sheng and the alternate mode exists, longer near the branch of major branch base portion, rare branch once more; Bottle obstructs: Dan Sheng, a life or 2-4 verticillate, sees very thin stigma more, and it is crooked slightly sometimes that its base portion is gradually narrow, waist is slightly wide, the top attenuates, and ultimate bottle stalk prolongation base portion is not hung and contracted, and is bent usually, presents asymmetric doleiform or " cattle horn shape ", 8.5-14 * 2.4-3 μ m;
Conidium: wall is smooth, pistac, obovate, ellipse, the wealthy circle in top, 2.5-4.8 * 1.8-3 μ m, 3-4 * 2-2.4 μ m usually.
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 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 sterilizations 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%.
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 Trichoderma with this.
The liquid fermentation production method of microorganism 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, 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 ℃, 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 T918 (Trichoderma pseudokoningii) that is used for the fungal diseases of plants biological control, and its preservation registration number is CGMCC NO.0442.
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.
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| CN 00109994 CN1337458A (en) | 2000-08-09 | 2000-08-09 | Microorganism (T918) and preparation method thereof |
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| CN 00109994 CN1337458A (en) | 2000-08-09 | 2000-08-09 | Microorganism (T918) and preparation method thereof |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104277984A (en) * | 2014-09-22 | 2015-01-14 | 华中农业大学 | Trichoderma koningiopsis ReTk1 bacterial strain and bacterial agent preparation method and application |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104277984A (en) * | 2014-09-22 | 2015-01-14 | 华中农业大学 | Trichoderma koningiopsis ReTk1 bacterial strain and bacterial agent preparation method and application |
| CN104277984B (en) * | 2014-09-22 | 2016-09-07 | 华中农业大学 | One intends healthy and free from worry Trichoderma spp. ReTk1 bacterial strain and bacterial preparation process and application |
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