CN1515666A - Method for screening pseudomonads for controlling blight - Google Patents
Method for screening pseudomonads for controlling blight Download PDFInfo
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- CN1515666A CN1515666A CNA200310124465XA CN200310124465A CN1515666A CN 1515666 A CN1515666 A CN 1515666A CN A200310124465X A CNA200310124465X A CN A200310124465XA CN 200310124465 A CN200310124465 A CN 200310124465A CN 1515666 A CN1515666 A CN 1515666A
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- strain
- antagonistic
- bacterial wilt
- pseudomonas
- screening
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Abstract
The method for screening pseudomonas for controlling bacterial wilt includes the following steps: (a). collecting root, stem and root medium soil of disease plant and health plant in the field with bacterial wilt; (2). separating strains; (3). utilizing Petri dish nutual antagonistic test between separated strains and pathogenicity test of separated strains to identify antagonistic strain and pathogenic strain; (4) using antagonistic rate and anti-antagonistic rate to analyze mutual retationship between antagonistic strain, pathogenic strain and others and select the antagonistic strains belonging to pseudomonas or bacillus; and (5). using the field controlling effect whis is whis is not less than 60% as reference selecting the antagonistic strain so as to obtain the effective strain.
Description
Technical field
The present invention relates to the screening method of a kind of bacterium of the bacterial wilt that is used to prevent and treat crop.The present invention is that application number is 00119018.0, the applying date is on October 11st, 2000, denomination of invention is divided an application for the application for a patent for invention of " pseudomonas stutzeri Zh 9944 and screening method thereof and prevent and treat the microbial preparation of bacterial wilt ".
Background technology
Fall ill in tobacco, tomato, peanut, eggplant, capsicum or the like crop by the bacterial wilt that the bacterial wilt pseudomonas causes, still find no the agricultural chemicals of effect up to now and prevent and treat method.
Summary of the invention
The purpose of this invention is to provide a kind of screening method that is used to prevent and treat the pseudomonas of bacterial wilt.
Realize that the object of the invention technical scheme is: the method that adopts the mutual antagonism screening model of plate to combine with directed screening, concrete steps are: root, stem and rhizosphere soil that 1. crop morbidity strain and healthy strain take place to gather in the piece of geographic field in bacterial wilt; 2. isolated strains from collected specimens; 3. by the mutual antagonistic effect of plate between isolated strains and the pathogenic test of isolated strains, distinguish antagonistic strain and pathogenic strains two classes; 4. analyze mutual relationship between antagonism bacterium, pathogenic bacteria and other classes with biological parameter antagonistic rate, anti-antagonistic rate, select the antagonistic strain that in division bacteria, belongs to false pseudomonas bacillus or genus bacillus; 5. carry out the multiple spot test of sub-district, field, being not less than 60% with field efficacy is that benchmark is selected the antagonistic strain of selecting, and resulting bacterial strain is effective strain, thereby has finished the screening of the pseudomonas that prevents and treats bacterial wilt.
Geographic field piece takes place above-mentioned bacterial wilt is that geographic field piece takes place for tobacco or bacterial wilt of tomato.
The present invention has positive effect: (1) does not have any agricultural chemicals up to now and can control because the bacterial wilt that causes because of Pseudomonas solanacearum is on the rise (comprising susceptible degree, crop and geographic rising and expansion).Method provided by the present invention can effectively be screened the bacterial isolates of control bacterial wilt.(2) in the application examples of the present invention, the pseudomonas of being screened that prevents and treats bacterial wilt is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center, and called after PSZh9944.PSZh9944 can be the former bacterium of susceptible crop inside and outside antagonism bacterial wilt; This bacterial strain has the ability of breeding in susceptible crop rhizosphere soils, and assembles and enter root tissue inside to crop root, upwards is transferred to stem.And be colonizated between the phloem parenchyma cells of root and stem, constitute so-called " antagonism defence line ", and form intergrowth relation with susceptible crop; PSZh9944 measures into corresponding relation to the size of Pseudomonas solanacearum antagonistic ability with the bacterium that inoculates PSZh9944 to the field.With PSZh9944 is that main component produces microbial pesticide, is the novel microorganism agricultural chemicals of a kind of efficient, nontoxic, safety, noresidue; This agricultural chemicals has very strong restraining effect to the bacterial wilt of plants such as the tobacco that causes because of Pseudomonas solanacearum, tomato, peanut, eggplant, capsicum, field efficacy reaches about 60～80%, applied thing and had prophylactic effect but also can make, and can be stimulated plant growth bacterial wilt evil.
Embodiment
(embodiment 1)
The method that adopts the mutual antagonism screening model of plate to combine with directed screening, concrete steps are:
1. from Greater, Changzhou, China Jiangsu Province bacterial wilt of tomato root, stem and the rhizosphere soil of morbidity strain and healthy strain take place in the piece of geographic field to gather in March, 1999;
2. isolated strains from collected specimens;
3. by the mutual antagonistic effect of plate between isolated strains and the pathogenic test of isolated strains, distinguish antagonistic strain and pathogenic strains two classes;
4. analyze mutual relationship between antagonism bacterium, pathogenic bacteria and other classes with biological parameters such as antagonistic rate, anti-antagonistic rate, select the antagonistic strain that in division bacteria, belongs to false pseudomonas bacillus (Pseudomonas spp.) or genus bacillus (Bacills spp.);
5. carry out the multiple spot test of sub-district, field, being not less than 60% with field efficacy is that benchmark is selected the antagonistic strain of selecting, resulting bacterial strain is effective strain, thereby finished the screening of the pseudomonas that prevents and treats bacterial wilt, the pseudomonas of being screened that prevents and treats bacterial wilt is deposited in China Committee for Culture Collection of Microorganisms common micro-organisms center on September 26th, 2000, it abbreviates CGMCC as, deposit number is NO.0494, and name is called Pseudomonas stutzeri (Pseudomonas stutzeri) Zh9944 (abbreviating the PSZh9944 bacterial strain as).
(application examples)
With glucose 1.5%, starch 2.0%, yeast powder 1.5%, peptone 0.4%, MgSO
4Be 0.05%, KH
2PO
4Be 0.05%, CaCO
3Be that 0.6% ratio is made substratum, pH value is 7.2～7.4 before the sterilization, at 70 milliliters of substratum of 250 milliliters of triangular flasks placements, the PSZh9944 of embodiment 1 gained is inoculated in this substratum.Shaking culture 48 hours is as seed liquor under 30 ℃, 220r.p.m, gets the 1000ml seed liquor and sends out in the alcohol jar the substratum of the above-mentioned same ratio that interior Sheng is 600 liters in 1 ton.At 30 ℃, to fermentor tank ventilate, stir culture 20～24 hours.After cultivate stopping, the rice chaff that in time with nutrient solution and granularity was 40 mesh sieves was with 1: 1～1: 3 mixed, and air seasoning to material moisture is 8～15%, and viable bacteria is measured and reaches 1000～6,000 hundred million CFU/g, and color and luster is that khaki color is to brown.
According to the nosogenesis of bacterial wilt, for the control of this type of disease, can received method can only be main prevention method in order to biological control, reach to stop with this and hide in ralstonia solanacearum invaded plants root and develop into disease.Therefore, use biotechnological formulation of the present invention opportunity should sow, heel in, after field planting and the field planting 20～60 days, 4～5 times altogether, total consumption is 2～2.5kg (300,000,000,000 CFU/g), be " solvent ", thalline is discharged with water in use, preventive effect can reach 60～80%.Concrete applicable cases sees Table 1 and table 2.
Table 1
| Test number | The crop title | Test site | The test time | For examination area (mu) | Treatment process | Handle | Contrast | Preventive effect % | ||
| Diseased plant rate % | Disease index | Diseased plant rate % | Disease index | |||||||
| 1 | Tomato | The changzhou | 2000 | 1.0 | Seed soaking+seedbed is sprinkled and is watered, altogether once.150g/ mu. | 10.13 | 10.13 | 14.96 | 16.51 | 38.6 |
| 2 | Tomato | The changzhou | 2000 | 1.0 | Seed soaking+seedbed is sprinkled and is watered, and heels in and waters root, twice totally.1.1kg/ mu. | 40 | 64 | 37.5 | ||
| 3 | Tomato | The changzhou | 2000 | 1.0 | Seed soaking+seedbed is sprinkled and is watered, and heels in and waters root, and root is watered in field planting, totally three times.1.5kg/ mu. | 40.0 | 87.0 | 54.0 | ||
| 4 | Tomato | The Nanchang | 1999 | 7.3 | Seed soaking+seedbed is sprinkled and is watered, heel in and water root, root is watered in field planting, waters root after the field planting, totally four times.2kg/ mu. | ???8.7 | ?4.8 | ?33 | ?19.7 | ?75.6 |
Table 2
| Test number | The crop title | Test site | The test time | For examination area (mu) | Treatment process | Handle | Contrast | Preventive effect % | ||
| Diseased plant rate % | Disease index | Diseased plant rate % | Disease index | |||||||
| 1 | Tomato | The changzhou | 1999 | 20 | Seed soaking+seedbed is sprinkled and is watered, heel in and water root, root is watered in field planting, waters root after the field planting, totally four times.2kg/ mu. | 4.2 | 40.7 | 89.7 | ||
| 2 | Tomato | The Nanchang | 2000 | 50 | The same. | 18.1 | 89.2 | 79.4 | ||
| 3 | Tobacco | Chenzhou, Hunan | 1999 | 10 | The same. | 44.67 | 22.57 | ?77.28 | ?51.01 | ?55.05 |
Claims (2)
1, a kind of screening method that is used to prevent and treat the pseudomonas of bacterial wilt, what it adopted is the method that the mutual antagonism screening model of plate combines with directed screening, and concrete steps are: root, stem and rhizosphere soil that 1. morbidity strain and healthy strain take place to gather in the piece of geographic field at tobacco, bacterial wilt of tomato; 2. isolated strains from collected specimens; 3. by the mutual antagonistic effect of plate between isolated strains and the pathogenic test of isolated strains, distinguish antagonistic strain and pathogenic strains two classes; 4. analyze mutual relationship between antagonism bacterium, pathogenic bacteria and other classes with biological parameter antagonistic rate, anti-antagonistic rate, select the antagonistic strain that in division bacteria, belongs to false pseudomonas bacillus or bacillus; 5. carry out the multiple spot test of sub-district, field, being not less than 60% with field efficacy is that benchmark is selected the antagonistic strain of selecting, and resulting bacterial strain is effective strain, thereby has finished the screening of the pseudomonas that prevents and treats bacterial wilt.
2, the screening method that is used to prevent and treat the pseudomonas of bacterial wilt according to claim 1 is characterized in that: geographic field piece takes place the bacterial wilt of step in 1. is that geographic field piece takes place for tobacco or bacterial wilt of tomato.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA200310124465XA CN1515666A (en) | 2000-10-11 | 2000-10-11 | Method for screening pseudomonads for controlling blight |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA200310124465XA CN1515666A (en) | 2000-10-11 | 2000-10-11 | Method for screening pseudomonads for controlling blight |
Related Parent Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB001190180A Division CN1159434C (en) | 2000-10-11 | 2000-10-11 | Pseudomonas stutzeri Zh9944 and its screening method and bacterial wilt preventing and controlling microbial prepn |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN1515666A true CN1515666A (en) | 2004-07-28 |
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| CNA200310124465XA Pending CN1515666A (en) | 2000-10-11 | 2000-10-11 | Method for screening pseudomonads for controlling blight |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1317953C (en) * | 2005-12-07 | 2007-05-30 | 南京农业大学 | Antago nistic fungus capable of preventing and treating continuous crops blight and its microbial organic fertilizer |
| CN100417942C (en) * | 2005-12-15 | 2008-09-10 | 浙江省农业科学院 | Tomato bacterial wilt resistance identification method |
| CN101864376A (en) * | 2010-02-10 | 2010-10-20 | 浙江省农业科学院 | Pseudomonas fluorescens strain, microbial inoculum and use thereof as seedling culture medium for controlling tomato bacterial wilt |
| CN101880672A (en) * | 2010-05-28 | 2010-11-10 | 华南农业大学 | Anti-plant-bacterial-wilt gene, preparation method and application thereof |
| CN102146350A (en) * | 2011-01-12 | 2011-08-10 | 南京农业大学 | Biocontrol strain 3YW8 for controlling bacterial wilt of ginger |
| CN101595892B (en) * | 2009-05-27 | 2011-08-17 | 南京农业大学 | PS mixture of compound biocontrol fungicide for preventing soil-borne diseases of vegetables |
| CN101914449B (en) * | 2010-02-08 | 2011-10-19 | 浙江省农业科学院 | Fungal inoculant for controlling bacterial wilt of tomato and preparation method thereof |
| CN101541948B (en) * | 2006-11-08 | 2012-03-28 | 日本曹达株式会社 | Microorganism capable of controlling plant diseases and plant disease-controlling agent using the microorganism |
| CN101148641B (en) * | 2007-08-06 | 2012-03-28 | 中国科学院南海海洋研究所 | High-flux fast screening method for antibiotics generated bacterium and device thereof |
| CN101928683B (en) * | 2010-01-01 | 2012-05-02 | 浙江大学 | Pseudomonas sp. strain and use thereof |
| CN102604857A (en) * | 2011-12-22 | 2012-07-25 | 中国农业科学院烟草研究所 | Biological control pseudomonas monteilii strain against tobacco mosaic virus (TMV) |
-
2000
- 2000-10-11 CN CNA200310124465XA patent/CN1515666A/en active Pending
Cited By (15)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1317953C (en) * | 2005-12-07 | 2007-05-30 | 南京农业大学 | Antago nistic fungus capable of preventing and treating continuous crops blight and its microbial organic fertilizer |
| CN100417942C (en) * | 2005-12-15 | 2008-09-10 | 浙江省农业科学院 | Tomato bacterial wilt resistance identification method |
| CN101541948B (en) * | 2006-11-08 | 2012-03-28 | 日本曹达株式会社 | Microorganism capable of controlling plant diseases and plant disease-controlling agent using the microorganism |
| CN101148641B (en) * | 2007-08-06 | 2012-03-28 | 中国科学院南海海洋研究所 | High-flux fast screening method for antibiotics generated bacterium and device thereof |
| CN101595892B (en) * | 2009-05-27 | 2011-08-17 | 南京农业大学 | PS mixture of compound biocontrol fungicide for preventing soil-borne diseases of vegetables |
| CN101928683B (en) * | 2010-01-01 | 2012-05-02 | 浙江大学 | Pseudomonas sp. strain and use thereof |
| CN101914449B (en) * | 2010-02-08 | 2011-10-19 | 浙江省农业科学院 | Fungal inoculant for controlling bacterial wilt of tomato and preparation method thereof |
| CN101864376B (en) * | 2010-02-10 | 2012-03-07 | 浙江省农业科学院 | Pseudomonas fluorescens strain, microbial inoculum and use thereof as seedling culture medium for controlling tomato bacterial wilt |
| CN101864376A (en) * | 2010-02-10 | 2010-10-20 | 浙江省农业科学院 | Pseudomonas fluorescens strain, microbial inoculum and use thereof as seedling culture medium for controlling tomato bacterial wilt |
| CN101880672B (en) * | 2010-05-28 | 2012-01-25 | 华南农业大学 | Anti-plant-bacterial-wilt gene, preparation method and application thereof |
| CN101880672A (en) * | 2010-05-28 | 2010-11-10 | 华南农业大学 | Anti-plant-bacterial-wilt gene, preparation method and application thereof |
| CN102146350A (en) * | 2011-01-12 | 2011-08-10 | 南京农业大学 | Biocontrol strain 3YW8 for controlling bacterial wilt of ginger |
| CN102146350B (en) * | 2011-01-12 | 2012-07-04 | 南京农业大学 | Biocontrol strain 3YW8 for controlling bacterial wilt of ginger |
| CN102604857A (en) * | 2011-12-22 | 2012-07-25 | 中国农业科学院烟草研究所 | Biological control pseudomonas monteilii strain against tobacco mosaic virus (TMV) |
| CN102604857B (en) * | 2011-12-22 | 2013-09-04 | 中国农业科学院烟草研究所 | Biological control pseudomonas monteilii strain against tobacco mosaic virus (TMV) |
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Open date: 20040728 |