CN1438312A - Method for producing Bicao anti-bateria spore using Bicao substrate - Google Patents
Method for producing Bicao anti-bateria spore using Bicao substrate Download PDFInfo
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- CN1438312A CN1438312A CN 02102975 CN02102975A CN1438312A CN 1438312 A CN1438312 A CN 1438312A CN 02102975 CN02102975 CN 02102975 CN 02102975 A CN02102975 A CN 02102975A CN 1438312 A CN1438312 A CN 1438312A
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- grass
- barnyard grass
- spore
- barnyard
- biocontrol microorganisms
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Abstract
The invention is a method of producing barnyard grass germproof spore by barnyard grass basic material. It separates to get barnyard grass germproof original germ plant from affected barnyard grass, after making nutrition on original germ plant, gets inoculant, cuts barnyard grass plant into small segments and dries, then mixes with carbon, nitrogen source and organic salt in proportion, disinfects at high temperature and high pressure and cools to make culture medium of the barnyard grass basic material. It inoculates the inoculant to the culture medium on the aseptic condition and cultivates the inoculant on the strictly disinfectant condition.
Description
[technical field]
The invention belongs to biological pesticide technical field.
[background technology]
It is popular that weeds biological control has at present become research, and various countries have filtered out many biological and ecological methods to prevent plant disease, pests, and erosion potentiality bacterium, are mainly fungi, and what have is developed to the weeds biological prevention and control agent.The barnyard grass grass is a main weeds in the rice field, over nearly 10 years, domestic and international many mechanisms have all carried out barnyard grass grass biological control research, separate from infected cockspur grass and to obtain the prominent navel spore (Exserohilum monoceras) of many pathogenic bacterias, especially pathogenic fungi wedge angle and interior navel child spore (Drechslera monoceras) is the most potential bacterial strain that is developed to the careless biological prevention and control agent of barnyard grass.The fungi biological prevention and control agent mainly is to utilize that its sporophore---spore infects target, makes its susceptible causing death, thereby reaches the purpose of control harmful organism.Therefore, the sporulation quantity of potentiality bacterium is one of principal element of this bacterium development and application prospect of decision.And present achievement in research also can't reach the actual promising result of applying of energy both at home and abroad, mainly is that sporulation quantity is on the low side.As international paddy rice the solid fermentation of E.monoceras bacterium spore on 22 kinds of agriculture materials produced the spore situation study, the maximum sporulation quantity of result is on leaf of Semen Maydis, it is 1.8 * 10 that every gram is done the leaf of Semen Maydis sporulation quantity
6Individual spore; The sporulation quantity of this bacterial strain on 27 kinds of agriculture materials studied by Vietnam, and maximum sporulation quantity appears at cavings, and it is every gram cavings 2.05 * 10 that second incubation is produced the spore sum
5Individual spore.
[summary of the invention]
The present invention seeks to: on the prior art basis, the further development research of production to barnyard grass grass biocontrol microorganisms spore, to overcome the low and defective that can't generally use of sporulation quantity, adopting barnyard grass grass be culture medium, and a kind of method of utilizing the careless biocontrol microorganisms spore of barnyard grass grass matrix production barnyard grass is provided.
Method with barnyard grass grass matrix is produced barnyard grass grass biocontrol microorganisms spore obtains barnyard grass grass biocontrol microorganisms original strain from the infected cockspur grass separation, makes inoculum after this strain culturing, is inoculated in the substratum, it is characterized in that comprising following processing step:
1) adopting the barnyard grass grass is culture medium, and barnyard grass grass plant is cut into segment, drying, standby.
2) by dried barnyard grass grass matrix: sucrose: KNO
3: K
2HPO
4: MgSO
4.7H
2O: CaCO
3=100: 3-9: 0.3-0.9: 0.2-0.6: 0.2-0.6: the ratio preparation of 0.2-0.6, soaked 24-36 hour, so at 121 ℃, 0.11MPa/cm
2High temperature, autoclave sterilization 15-30 minute make barnyard grass grass matrix substratum after the cooling, be used for inoculation, produce spore.
3) under aseptic condition, barnyard grass grass biocontrol microorganisms inoculum is inoculated in the substratum, be stirred well to and mix, evenly spread then, under 25-29 ℃ of constant temperature, dark, blowing air condition, cultivated 7-10 days, at 25-30 ℃ of normal temperature, under the strict sterilization envrionment conditions, cultivate and to gather in the crops spore in 10-15 days subsequently.
The barnyard grass grass that this production method adopted, its vitality and growing ability are extremely strong, and artificial growth is convenient, so the raw material cheapness, is easy to get, and production technique is simple, but large-scale development production, its sporulation quantity height, the 3.75-33.07 that reaches external like product is doubly.
[specific embodiments]
Separate to obtain barnyard grass grass biocontrol microorganisms original strain from infected cockspur grass, described barnyard grass grass biocontrol microorganisms bacterial strain is the prominent navel spore (Exserohilum monoceras) of pathogenic fungi wedge angle or interior navel child spore (Drechsleramonoceras) or the curved spore (Curvularia lunata) of lattice interlinkage spore (Alternaria alternata) mutually.Concrete separation method is conventional known technology, does not repeat them here.
The preparation of barnyard grass grass matrix substratum: the neat mud of barnyard grass grass plant of milk stage is cut off, and comprises stem, leaf and seed, and cuts into the long segment of 0.1-0.5cm, shines or dries to moisture standby below 14%.Make barnyard grass grass matrix substratum by aforementioned proportioning and method, preferably add a small amount of vitamins C in the preparation.
The preparation of barnyard grass grass biocontrol microorganisms inoculum: barnyard grass grass biocontrol microorganisms original strain is seeded on the PDA substratum, cultivates 6-8 days under 25-29 ℃ of constant temperature, dark condition, forms the one-level inoculum, and the one-level inoculum can be directly used in a small amount of product spore inoculation.As be used for scale operation, then need cultivate the two-stage inoculation body, two kinds of methods are specifically arranged: 1) the one-level inoculum is seeded in improvement Fries liquid nutrient medium or potato, the liquid sucrose substratum, carries out obtaining mycelia in fermentation culture 15-20 days, make the two-stage inoculation body after mycelia is smashed to pieces.2) the one-level inoculum is seeded on the barnyard grass grass-seed grain of high temperature, autoclave sterilization, is preferably yellow maturity-ripening stage barnyard grass seed, under 25-29 ℃ of constant temperature, dark condition, cultivated 10-14 days, promptly get the two-stage inoculation body after waiting to produce a large amount of spores and mycelium.Described improvement Fries liquid nutrient medium proportioning is: sucrose 30 grams, winestone acid amide 5.0 grams, NH
4NO
31.0 gram, K
2HPO
41.0 gram, MgSO
40.5 gram, NaCL0.1 gram, CaCL
20.1 1000 milliliters of grams, hydrolysed casein 0.5 gram, yeast extract (yeast extraact) 1.0 grams, sterilized water.Described potato, liquid sucrose substratum are: 1000 milliliters of peeling potato 200-300 grams, sucrose 10-20 gram, sterilized water, and again through high temperature, autoclave sterilization.Sucrose wherein can replace with glucose.
The two-stage inoculation body is inoculated in the solid medium of the cooled barnyard grass grass of above-mentioned sterilization matrix, organic salt and trace element, fully stir and make inoculum and medium component uniform mixing, evenly spread out then in the aluminium dish of sterilizing or disk or dustpan, more than operation needs to carry out under aseptic condition.25-29 ℃ of constant temperature, dark, have under the air conditions and to cultivate 7-10 days, fully stirred once in every 2-3 days, make bacterium fast, evenly growth.Subsequently can be at 25-30 ℃ of normal temperature, under the environment of strict sterilization, cultivate.Can gather in the crops spore in 10-15 days.As adopt the washing method results, and after the culture medium filter solid carbon dioxide after the results divides, can directly be used as second incubation, can produce a large amount of spores equally.Under the few situation of cultivation amount, be more or less the same with inoculation of bacterium piece and spore suspension effect of inoculation, but when large-scale inoculation is cultivated, be good with the spore suspension inoculation.
Claims (8)
1. produce the method for barnyard grass grass biocontrol microorganisms spore with barnyard grass grass matrix, separate obtaining barnyard grass grass biocontrol microorganisms original strain from infected cockspur grass, make inoculum after this strain culturing, be inoculated in the substratum, it is characterized in that comprising following processing step:
1) adopting the barnyard grass grass is culture medium, and barnyard grass grass plant is cut into segment, drying, standby,
2) by dried barnyard grass grass matrix: sucrose: KNO
3: K
2HPO
4: MgSO
4.7H
2O: CaCO
3=100: 3-9: 0.3-0.9: 0.2-0.6: 0.2-0.6: the ratio preparation of 0.2-0.6, soaked 24-36 hour, then at 121 ℃, 0.11MPa/cm
2High temperature, autoclave sterilization 15-30 minute make barnyard grass grass matrix substratum after the cooling, and be used for inoculation, produce spore,
3) under aseptic condition, barnyard grass grass biocontrol microorganisms inoculum is inoculated in the substratum, be stirred well to and mix, evenly spread then, under 25-29 ℃ of constant temperature, dark, blowing air condition, cultivated 7-10 days, at 25-30 ℃ of normal temperature, under the strict sterilization envrionment conditions, cultivate and to gather in the crops spore in 10-15 days subsequently.
2. method of producing barnyard grass grass biocontrol microorganisms spore with barnyard grass grass matrix as claimed in claim 1 is characterized in that being added with a spot of vitamins C in the barnyard grass grass substratum.
3. method of producing barnyard grass grass biocontrol microorganisms spore with the careless matrix of barnyard grass as claimed in claim 1 is characterized in that barnyard grass grass biocontrol microorganisms bacterial strain is the prominent navel spore (Exserohilum monoceras) of pathogenic fungi wedge angle or interior navel child spore (Drechslera monoceras) or mutual lattice interlinkage spore (Alternaria alternata) or curved spore (Curvularia lunata).
4. method of producing barnyard grass grass biocontrol microorganisms spore with barnyard grass grass matrix as claimed in claim 1, it is characterized in that the preparation of barnyard grass grass biocontrol microorganisms inoculum: barnyard grass grass biocontrol microorganisms original strain is seeded on the PDA substratum, under 25-29 ℃ of constant temperature, dark condition, cultivated 6-8 days, form the one-level inoculum, again the one-level inoculum is seeded in improvement Fries liquid nutrient medium or potato, the liquid sucrose substratum, carry out obtaining mycelia in fermentation culture 15-20 days, make the two-stage inoculation body after mycelia is smashed to pieces.
5. method of producing barnyard grass grass biocontrol microorganisms spore with barnyard grass grass matrix as claimed in claim 1, it is characterized in that the preparation of barnyard grass grass biocontrol microorganisms inoculum: barnyard grass grass biocontrol microorganisms original strain is seeded on the PDA substratum, under 25-29 ℃ of constant temperature, dark condition, cultivated 6-8 days, form the one-level inoculum, again the one-level inoculum is seeded on the barnyard grass grass-seed grain of high temperature, autoclave sterilization, under 25-29 ℃ of constant temperature, dark condition, cultivated 10-14 days, after waiting to produce a large amount of spores and mycelium, promptly get the two-stage inoculation body.
6. method of producing barnyard grass grass biocontrol microorganisms spore with barnyard grass grass matrix as claimed in claim 4 is characterized in that described improvement Fries liquid nutrient medium proportioning is: sucrose 30 grams, winestone acid amide 5.0 grams, NH
4NO
31.0 gram, K
2HPO
41.0 gram, MgSO
40.5 gram, NaCL0.1 gram, CaCL
20.1 1000 milliliters of grams, hydrolysed casein 0.5 gram, yeast extract (yeast extraact) 1.0 grams, sterilized water.
7. method of producing barnyard grass grass biocontrol microorganisms spore with barnyard grass grass matrix as claimed in claim 4, it is characterized in that described potato, liquid sucrose substratum are: 1000 milliliters of peeling potato 200-300 grams, sucrose 10-20 gram, sterilized water, again through high temperature, autoclave sterilization.
8. as claim 4 or 7 described methods of producing barnyard grass grass biocontrol microorganisms spore, it is characterized in that the sucrose in described potato, the liquid sucrose cultivation can replace with glucose with barnyard grass grass matrix.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN 02102975 CN1219051C (en) | 2002-02-10 | 2002-02-10 | Method for producing Bicao anti-bateria spore using Bicao substrate |
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|---|---|---|---|
| CN 02102975 CN1219051C (en) | 2002-02-10 | 2002-02-10 | Method for producing Bicao anti-bateria spore using Bicao substrate |
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| CN1438312A true CN1438312A (en) | 2003-08-27 |
| CN1219051C CN1219051C (en) | 2005-09-14 |
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|---|---|---|---|
| CN 02102975 Expired - Fee Related CN1219051C (en) | 2002-02-10 | 2002-02-10 | Method for producing Bicao anti-bateria spore using Bicao substrate |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102428965A (en) * | 2011-09-22 | 2012-05-02 | 中国水稻研究所 | Specialized form spore powder of helminthosporium carposaprum echinochloa crusgalli |
| CN105886410A (en) * | 2016-04-27 | 2016-08-24 | 山东省农业科学院植物保护研究所 | Myrothecium culture medium containing barnyard grass and preparation method thereof |
| CN108935522A (en) * | 2018-07-05 | 2018-12-07 | 安徽袁粮水稻产业有限公司 | A kind of compound drug and preparation method thereof for preventing and treating barnyard grass |
| CN109735457A (en) * | 2019-02-21 | 2019-05-10 | 华南农业大学 | One plant of mutagenesis Infected barnyardgrass and its application in prevention and treatment barnyard grass |
| CN111088172A (en) * | 2020-03-06 | 2020-05-01 | 中国水稻研究所 | Method for enhancing weeding toxicity of helminthosporium peregrinum spores |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN100403909C (en) * | 2006-04-21 | 2008-07-23 | 中国水稻研究所 | Production method of induced anti-heterogenous fungus protein agricultural chemical for promoting crop growth based on moss slag |
-
2002
- 2002-02-10 CN CN 02102975 patent/CN1219051C/en not_active Expired - Fee Related
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102428965A (en) * | 2011-09-22 | 2012-05-02 | 中国水稻研究所 | Specialized form spore powder of helminthosporium carposaprum echinochloa crusgalli |
| CN105886410A (en) * | 2016-04-27 | 2016-08-24 | 山东省农业科学院植物保护研究所 | Myrothecium culture medium containing barnyard grass and preparation method thereof |
| CN108935522A (en) * | 2018-07-05 | 2018-12-07 | 安徽袁粮水稻产业有限公司 | A kind of compound drug and preparation method thereof for preventing and treating barnyard grass |
| CN109735457A (en) * | 2019-02-21 | 2019-05-10 | 华南农业大学 | One plant of mutagenesis Infected barnyardgrass and its application in prevention and treatment barnyard grass |
| CN109735457B (en) * | 2019-02-21 | 2020-04-14 | 华南农业大学 | Mutant eurotium cristatum and application thereof in preventing and controlling barnyard grass |
| CN111088172A (en) * | 2020-03-06 | 2020-05-01 | 中国水稻研究所 | Method for enhancing weeding toxicity of helminthosporium peregrinum spores |
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| Publication number | Publication date |
|---|---|
| CN1219051C (en) | 2005-09-14 |
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Granted publication date: 20050914 Termination date: 20130210 |