CN105567600A - Pathogen verticillium antagonistic bacterium and application thereof - Google Patents
Pathogen verticillium antagonistic bacterium and application thereof Download PDFInfo
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Abstract
The invention discloses a pathogen verticillium antagonistic bacterium and application thereof, and belongs to the technical field of biological control over crop diseases. Bacillus pumilus Y106 is adopted as the pathogen verticillium antagonistic bacterium, the preservation number is CCTCC NO:M2014423, the preservation date is September 17, 2014, the preservation place is China Center for Type Culture Collection (CCTCC), and the preservation address is Wuhan University in Wuhan China. The bacterial strain is Gram-positive, mesophilic and aerobiotic and can resist the high temperature of 65 DEG C; the bacterial strain comprises the main morphological characteristics that the bacterial strain is shaped like a thin rod, the two ends are bluntly round, and the bacterial strain mostly exists in a single mode or in a chain-arranged mode. The pathogen verticillium antagonistic bacterium has the antagonistic effect on pathogen verticillium-verticillium dahliae, can inhibit germination of pathogen spores and can inhibit paroxysm of cotton verticillium wilt to a certain degree.
Description
Technical field
The present invention relates to a kind of verticillium dahliae Antagonistic Fungi, also relate to the application of this Antagonistic Fungi simultaneously, belong to crop pest technical field of biological control.
Background technology
Cotton is the important cash crop of China, and there is plantation most of provinces and regions.Cotton verticillium wilt (Verticilliumwilt) is by the microbial a kind of soil-borne disease of verticillium, and in the world, all there is distribution each cotton production areas, and in the trend day by day spread.This disease infestation cotton fibrovascular system, have the title of " cancer of cotton ", and the other plants such as fruit, beans, greengrocery can be made susceptible, causes quality of agricultural product and production declining.How effectively to control the generation of cotton verticillium wilt, alleviate its harm thus ensure that the high and stable yields of cotton have caused the extensive concern of people.
Verticillium dahliae has the features such as adaptable, host range is wide, Virulence is obvious, route of transmission is various, the mechanism of action is complicated.Owing to lacking the anti-source material of high-quality, breeding for disease resistance work is restricted, and there is no the high resistance cotton variety for this disease at present.And conventional chemical fungicides lacks specificity, the generation of drug resistance strain can be caused, cause environmental pollution.Therefore, develop the strong chemical agent of high-efficiency low-toxicity, specificity and biological control agent extremely urgent.
Utilize antagonistic microbe to carry out biological control to be considered to prevent and treat the most potential method of verticillium.In recent years, successively find that some have the bacterium of antagonistic action, fungi and actinomycetes to cotton verticillium wilt, study in antagonistic bacterium maximum be genus bacillus, for preventing and treating genus bacillus (Bacillussubtilis) 41B-1 (deposit number: CGMCCNo.5582) of cotton verticillium wilt disclosed in the patent of invention of notification number CN102443559B, the sub-genus bacillus (Bacillusaxarquiensisi) of Amado Azar base disclosed in the patent of invention of notification number CN104082344A etc.
Summary of the invention
The object of this invention is to provide a kind of verticillium wilt pathogen Antagonistic Fungi.
Meanwhile, the present invention also provides a kind of purposes of verticillium wilt pathogen Antagonistic Fungi.
Finally, the present invention also provides a kind of biocontrol fungicide for preventing and treating crop verticillium.
In order to realize above object, the technical solution adopted in the present invention is:
Verticillium wilt pathogen Antagonistic Fungi, its name is called bacillus pumilus (Bacilluspumilus) Y106, deposit number: CCTCCNO:M2014423, preservation date: on September 17th, 2014, depositary institution: China typical culture collection center (CCTCC), preservation address: Wuhan, China. Wuhan University (Luojiashan, Wuchang, Wuhan City, Hubei Province Wuhan University preservation center).
Bacillus pumilus (Bacilluspumilus) Y106 is isolated a kind of Cotton rhizosphere bacterium from pedotheque, is Gram-positive, addicted to warm, aerobic, can tolerate 65 DEG C of high temperature.Main morphological features is: thin rod shape, the blunt circle in two ends, exists mainly with single or catenation form.Bacillus pumilus Y106 bacterium colony on LB flat board is opaque, is creamy white, circular, has stickiness, wetting, smooth; Cultivate more than 72 hours, colony edge starts shrinkage, indentation.
The cultural method of verticillium wilt pathogen Antagonistic Fungi:
1) test tube slant is cultivated
Adopt modified form LB substratum, cultivate 1 ~ 3 day at 28 ~ 32 DEG C, obtain test tube kind.Modified form LB culture medium prescription is: yeast extract 1 ~ 3%, peptone 0.5 ~ 2%, glucose 1 ~ 4%, NaCl1 ~ 3%, agar 1.8 ~ 2%; PH7.0.
2) culture dish is streak culture
Adopt above-mentioned modified form LB substratum, streak inoculation bacillus pumilus (Bacilluspumilus) Y106, cultivate 1 ~ 2 day at 28 ~ 32 DEG C after sealed membrane sealing, obtain the single bacterium colony being used for fluid enlargement culture.
3) fluid enlargement culture
By the streak culture single colony inoculation obtained in liquid nutrient medium, under temperature 25 ~ 37 DEG C, rotating speed 200 ~ 300rpm, shaking culture 2 ~ 5 days, to obtain final product.Liquid culture based formulas is: yeast extract 1 ~ 3%, peptone 0.5 ~ 2%, glucose 1 ~ 4%, NaCl1 ~ 3%; PH7.0.
The purposes of verticillium wilt pathogen Antagonistic Fungi, comprise: the application of verticillium wilt pathogen Antagonistic Fungi (CCTCCNO:M2014423) in the biocontrol fungicide for the preparation of control crop (especially cotton) verticillium, or verticillium wilt pathogen Antagonistic Fungi (CCTCCNO:M2014423) is preparing the application in microbial fertilizer (especially microbial organic fertilizer).
The purposes of verticillium wilt pathogen Antagonistic Fungi, also comprise: the application of verticillium wilt pathogen Antagonistic Fungi (CCTCCNO:M2014423) in control crop (especially cotton) verticillium, or verticillium wilt pathogen Antagonistic Fungi (CCTCCNO:M2014423) is promoting the application in crop (especially cotton) plant strain growth.
For preventing and treating biocontrol fungicide or the microbial fertilizer of crop verticillium, it comprises verticillium wilt pathogen Antagonistic Fungi (CCTCCNO:M2014423).
Beneficial effect of the present invention:
In the present invention, verticillium wilt pathogen Antagonistic Fungi is bacillus pumilus (Bacilluspumilus) Y106 (CCTCCNO:M2014423), it is isolated a kind of Cotton rhizosphere bacterium from pedotheque, for Gram-positive, addicted to warm, aerobic, 65 DEG C of high temperature can be tolerated.Main morphological features is: thin rod shape, the blunt circle in two ends, exists mainly with single or catenation form.Bacillus pumilus Y106 bacterium colony on LB flat board is opaque, is creamy white, circular, has stickiness, wetting, smooth; Cultivate more than 72 hours, colony edge starts shrinkage, indentation.
In the present invention, verticillium wilt pathogen Antagonistic Fungi (CCTCCNO:M2014423) has antagonistic action to verticillium pathogenic bacteria verticillium dahliae (Verticiliumdahliae), pathogenic bacteria spore germination can be suppressed, suppress cotton verticillium wilt outbreak to a certain extent.
Preservation proves and survival proves explanation
Preservation strain: bacillus pumilus (Bacilluspumilus) Y106, deposit number: CCTCCNO:M2014423, preservation date: on September 17th, 2014, depositary institution: China typical culture collection center (CCTCC), preservation address: Wuhan, China. Wuhan University (Luojiashan, Wuchang, Wuhan City, Hubei Province Wuhan University preservation center).
Accompanying drawing explanation
Fig. 1 is the opposite culture design sketch of bacillus pumilus Y106 and verticillium dahliae 991 in test example 1;
Fig. 2 be in test example 2 bacillus pumilus Y106 bacterium liquid to the inhibition figure of verticillium dahliae gfp77 spore germination;
Fig. 3 is the incidence of cotton seedling under different treatment in test example 3.
Embodiment
Following embodiment is only described in further detail the present invention, but does not form any limitation of the invention.
Embodiment 1
The Isolation and screening method of verticillium wilt pathogen Antagonistic Fungi, comprises the following steps:
1), after cotton harvesting, from Henan, the material that 53 parts, Cotton rhizosphere soil is separated as Antagonistic Fungi is got in each cotton planting district;
2) select the LB substratum of improvement, adopt dilution spread flat band method to carry out strain separating to 53 parts of earth materials, be divided into from acquisition 537 strain bacterium;
3) adopt dull and stereotyped opposite culture method to screen being separated the bacterial strain obtained, wherein 14 strain bacteriums have obvious antagonistic action to cotton verticillium wilt pathogenic bacteria, and the bacterial strain antagonistic effect being numbered Y106 is the most obvious, and bacteriostasis rate is about 30%.
1, Antagonistic Fungi morphological observations
Bacillus pumilus (Bacilluspumilus) Y106 bacterium colony on LB flat board is opaque, is creamy white, circular, has stickiness, wetting, smooth; Cultivate more than 72 hours, colony edge starts shrinkage, indentation.
2, the qualification classification of Antagonistic Fungi
1) cellular form and physical and chemical inspection
Bacillus pumilus (Bacilluspumilus) Y106 is Gram-positive, addicted to warm, aerobic, can tolerate 65 DEG C of high temperature.Main morphological features is: thin rod shape, the blunt circle in two ends, exists mainly with single or catenation form.
2) 16SrRNA gene sequencing result
Purifying bacterial strain Y106, gets single bacterium colony streak culture, and serve Hai Shenggong biotechnology service company and check order, sequencing result is shown in SEQ ID NO.3, the homology of result display and bacillus pumilus (Bacilluspumilus).
Sequencing primer is as follows:
Forward primer 27F:5 '-AGAGTTTGATCCTGGCTCAG-3 ' (as shown in SEQIDNO.1),
Reverse primer 1495R:5 '-CTACGGCTACCTTGTTACGA-3 ' (as shown in SEQIDNO.2).
The cultivation of verticillium wilt pathogen Antagonistic Fungi:
1) test tube slant is cultivated
Adopt modified form LB substratum, cultivate 2 days at 30 DEG C, obtain test tube kind.Modified form LB culture medium prescription is: yeast extract 2%, peptone 1%, glucose 2.5%, NaCl2%, agar 1.9%; PH7.0.
2) culture dish is streak culture
Adopt above-mentioned modified form LB substratum (diameter 9mm culture dish), streak inoculation bacillus pumilus (Bacilluspumilus) Y106, cultivate 1 day at 30 DEG C after sealed membrane sealing, obtain the single bacterium colony being used for fluid enlargement culture.
3) fluid enlargement culture
By the streak culture single colony inoculation obtained to (500mL triangular flask, every bottled 200mL) in liquid nutrient medium, under temperature 32 DEG C, rotating speed 250rpm, shaking culture (shaking table) 3 days, to obtain final product.Liquid culture based formulas is: yeast extract 2%, peptone 1%, glucose 2.5%, NaCl2%; PH7.0.
China typical culture collection center preservation is delivered to by cultivating bacillus pumilus (Bacilluspumilus) Y106 obtained, deposit number: CCTCCNO:M2014423, preservation date: on September 17th, 2014, preservation address: Wuhan, China. Wuhan University (Luojiashan, Wuchang, Wuhan City, Hubei Province Wuhan University preservation center).
Embodiment 2
The application of verticillium wilt pathogen Antagonistic Fungi (CCTCCNO:M2014423) in the biocontrol fungicide for the preparation of control cotton verticillium wilt.Be specially: get verticillium wilt pathogen Antagonistic Fungi enlarged culturing, collect thalline, after sterilized water dilution, obtain liquid microbial inoculum.Strain expanded culture and microorganism collection method are prior art, repeat no more herein.
The application of verticillium wilt pathogen Antagonistic Fungi (CCTCCNO:M2014423) in control cotton verticillium wilt.Be specially: cotton seeds is placed in above-mentioned liquid microbial inoculum, seed soaking, then take out seed and dry.Bacteria suspension concentration and seed soaking operation are prior art, repeat no more herein.
Embodiment 3
Verticillium wilt pathogen Antagonistic Fungi (CCTCCNO:M2014423) is preparing the application in microbial organic fertilizer.Be specially: get verticillium wilt pathogen Antagonistic Fungi enlarged culturing, collect thalline, after sterilized water dilution, obtain liquid microbial inoculum.Method is with embodiment 2.
Verticillium wilt pathogen Antagonistic Fungi (CCTCCNO:M2014423) is promoting the application in cotton plants growth.Be specially: get above-mentioned liquid microbial inoculum, fill with root (or spraying) at cotton in seedling stage (or initial bloom stage).
Embodiment 4
For preventing and treating the biocontrol fungicide of crop verticillium, preparation process comprises: get verticillium wilt pathogen Antagonistic Fungi (CCTCCNO:M2014423) enlarged culturing, collects thalline, obtains liquid microbial inoculum after sterilized water dilution.The preparation process of microbial fertilizer is the same.
Test example
1, the dull and stereotyped dual test of bacillus pumilus Y106 and verticillium dahliae (Verticiliumdahliae) 991
1) preparation of pathogenic bacteria spore suspension
Cotton verticillium wilt pathogenic bacteria verticillium dahliae (Verticiliumdahliae) 991 (being so kind as to give by Institute of Agricultural Product Processing, Chinese Academy of Agricultural Sc) is inoculated in solid culture on PDA flat board, be paved with after flat board until bacterium colony, on super clean bench by fungal spore scraping in sterilized water, cross and filter mycelia, adopt blood counting chamber direct census spore quantity, then spore suspension concentration is adjusted to 5 × 10
7individual/mL.
2) preparation of verticillium wilt pathogen Antagonistic Fungi suspension
After bacillus pumilus (Bacilluspumilus) Y106 fluid enlargement culture, collected by centrifugation thalline, suspends with sterilized water, calculates bacteria suspension concentration by the method for plate culture count, regulates bacterial concentration to 3.77 × 10
7cfu/mL.
3) flat plate mutual confrontation test method
Super clean bench draws 2.5 μ L pathogenic bacteria spore suspensions with liquid-transfering gun and is inoculated in PDA substratum, be inverted in 25 DEG C of incubators and cultivate, when fungal growth is about 3.5cm to diameter, at distance 1.5cm (1 ~ 1.5cm) place, fungi edge inoculation verticillium wilt pathogen Antagonistic Fungi suspension 2.5 μ L, continue in 25 DEG C of incubators to be inverted and cultivate, observe fungal growth situation, three parallel twice repetitions are established in test.
Being calculated as follows of bacteriostasis rate and bacteriostatic diameter:
Bacteriostasis rate (%)=[(contrast fungal growth diameter-process fungal growth diameter)/contrast fungal growth diameter] × 100%;
Bacteriostatic diameter (cm)=contrast fungal growth diameter-process fungal growth diameter.
4) test-results
The dull and stereotyped opposite culture of bacillus pumilus (Bacilluspumilus) Y106 and cotton verticillium wilt pathogenic bacteria verticillium dahliae (Verticiliumdahliae) 991 is tested and carries out Continuous Observation, result display bacillus pumilus Y106 has obvious restraining effect (see Fig. 1 and following table 1) to verticillium dahliae 991, show that bacillus pumilus Y106 has certain antagonistic action to cotton verticillium wilt pathogenic bacteria, can as potentiality biocontrol strain.
In Fig. 1, a is verticillium dahliae 991 bacterium colony, and b is verticillium dahliae 991 and bacillus pumilus Y106 opposite culture; In Fig. 1 b, A is bacillus pumilus Y106, B is verticillium dahliae 991.
The dull and stereotyped dual test result of table 1 bacillus pumilus Y106 and verticillium dahliae 991
2, bacillus pumilus Y106 is to verticillium dahliae (Verticiliumdahliae) gfp77 spore germination inhibition test
Cotton verticillium wilt pathogenic bacteria verticillium dahliae (Verticiliumdahliae) gfp77 is the pCH-sGFP vector strain of verticillium dahliae (Verticiliumdahliae) 991, fluorescent signal is strong, with verticillium dahliae 991 grow and in virulence without significant difference, be so kind as to give by Institute of Agricultural Product Processing, Chinese Academy of Agricultural Sc.Select gfp77 to replace 991 and carry out spore germination Inhibition test, pathogenic bacteria and other miscellaneous bacterias can be differentiated fast by UV-irradiation, effectively reduce the interference of miscellaneous bacteria in data statistics.
1) preparation of pathogenic bacteria spore suspension
The preparation of verticillium dahliae (Verticiliumdahliae) gfp77 spore suspension is with verticillium dahliae 991 in test example one, and concentration is 5 × 10
6individual/mL.
2) preparation of verticillium wilt pathogen Antagonistic Fungi suspension
The preparation of bacillus pumilus (Bacilluspumilus) Y106 suspension, with test example one, makes 9.43 × 10 respectively
6cfu/mL, 1.72 × 10
7cfu/mL, 2.77 × 10
7cfu/mL3 concentration gradient.
3) preparation of sterile soil
Take soil 30g, put into wide mouth glass bottle, be cooled to room temperature after adding sealed membrane sterilizing for subsequent use.
4) test method
Get 5mL pathogenic bacteria spore suspension, mix with the verticillium wilt pathogen Antagonistic Fungi suspension of 5mL different concns respectively, load in throat spray, on Bechtop, evenly spray sterilized soil (as test group), add sealed membrane to seal up, cultivate 5 days in 25 DEG C of incubators; Contrast 1 group for spraying 10mL sterilized water in sterilized soil; Contrasting 2 groups is the pathogenic bacteria spore suspension and the sterilized water that spray the mixing of 10mL geometric ratio.After 5 days, from test group and control group, get 10g soil sample respectively, be mixed with soil supension with 90mL sterilized water.Adopt dilution spread flat band method, by soil supension with 10
-1for gradient doubling dilution, coating paraxin, the dual anti-flat board of Totomycin.The flat-plate inverted of coating is placed in 25 DEG C of incubators and cultivates 3 days (3 ~ 4 days), observed and recorded colony growth situation, statistics colony growth number.Three parallel twice repetitions are established in test.
5) test-results
As shown in Fig. 2 and following table 2, when pathogenic bacteria spore suspension concentration is constant, along with the increase of bacillus pumilus Y106 suspension concentration, verticillium dahliae gfp77 colony count is minimizing trend.In Fig. 2, a is contrast 1 group, and b is contrast 2 groups, and c ~ e is different concns verticillium wilt pathogen Antagonistic Fungi suspension process test group, and in c ~ e, Antagonistic Fungi suspension concentration is followed successively by 9.43 × 10
6cfu/mL, 1.72 × 10
7cfu/mL, 2.77 × 10
7cfu/mL.
The number of verticillium dahliae gfp77 bacterium colony after the process of table 2 different concns bacillus pumilus Y106 bacterium liquid
3, bacillus pumilus Y106 tests the biological and ecological methods to prevent plant disease, pests, and erosion (potted plant) of cotton verticillium wilt
1) preparation of pathogenic bacteria spore suspension and verticillium wilt pathogen Antagonistic Fungi suspension
Preparation process is with test example one, and wherein the concentration of verticillium dahliae (Verticiliumdahliae) 991 spore suspension is 1 × 10
7the concentration of individual/mL, bacillus pumilus (Bacilluspumilus) Y106 suspension is 1.72 × 10
7cfu/mL.
2) preparation of sterile soil
Get 500cm
3put into wide mouth glass bottle, be cooled to room temperature after adding sealed membrane sterilizing for subsequent use.
3) preparation of cotton seedling
Cotton kind in plasticine alms bowl, the strain of every alms bowl 2, when growing about 15 days (rough leaf is launched completely) in cotton incubator, taken out from native alms bowl by cotton seedling, at the tip of a root, place cuts off 2cm, for subsequent use.
4) biological and ecological methods to prevent plant disease, pests, and erosion test
Get 30mL pathogenic bacteria spore suspension and verticillium wilt pathogen Antagonistic Fungi suspension respectively, mixing, to be evenly sprayed on sterilized soil with throat spray and to stir, then soil is sub-packed in disposable plastic soil alms bowl; To cut the cotton replanting of root to native alms bowl, the strain of every alms bowl 2, is placed in cotton incubator, cultivates 20 days at temperature 20 DEG C, according to following grade scale statistics cotton incidence.1 group of contrast is established in test, and control group is that the mixed solution of 30mL pathogenic bacteria spore suspension and 30mL sterilized water sprays sterile soil, and test repeats 6 times altogether.
Cotton verticillium wilt to be fallen ill level grade scale seedling stage:
0 grade: be good for seedling, without symptom;
1 grade: 1-2 sheet cotyledon morbidity, cotyledons turn yellow, deliquescing, the not aobvious symptom of true leaf;
2 grades: cotyledon and 1 true leaf performance symptom;
3 grades: 2 true leaf performance symptom;
4 grades: all blade performance symptom, until leaf abscission, the top heart is withered.
Being calculated as follows of sickness rate and disease index:
Sickness rate (%)=morbidity strain number/investigate total strain number × 100%
Disease index=Σ (morbidity level typical value × diseased plant number at different levels) × 100/ (investigating total strain number × superlative degree morbidity typical value)
5) biological and ecological methods to prevent plant disease, pests, and erosion test-results
As shown in Fig. 3 and following table 3, after the process of bacillus pumilus Y106 bacterium liquid, cotton in seedling stage verticillium disease disease index and sickness rate obviously reduce, and 6 times the average preventive effect of revision test is 32.5%.In Fig. 3, a is control group, and b is process test group.
Table 3 bacillus pumilus Y106 biological and ecological methods to prevent plant disease, pests, and erosion test-results
Note: in table, data are 6 revision test mean values, same letter represents difference not significantly (P > 0.05) thereafter.
Test-results shows, bacillus pumilus (Bacilluspumilus) Y106 has good antagonistic effect and prevention effect to cotton verticillium wilt pathogenic bacteria verticillium dahliae (Verticiliumdahliae) 991, possesses good application and development prospect.At biocontrol microorganisms in pathogenic bacteria (verticillium dahliae) spore germination inhibition test, measure for getting rid of the elite Verticiliumdahliae991 mutant strain Verticiliumdahliaegfp77 carrying gfp gene and hygromycin gene that gets of miscellaneous bacteria interference, result display biocontrol strain reaches biocontrol effect by suppressing pathogenic bacteria spore germination.
Claims (7)
1. verticillium wilt pathogen Antagonistic Fungi, is characterized in that: the name of verticillium wilt pathogen Antagonistic Fungi is called bacillus pumilus (Bacilluspumilus) Y106, deposit number: CCTCCNO:M2014423.
2. the application of verticillium wilt pathogen Antagonistic Fungi as claimed in claim 1 in the biocontrol fungicide for the preparation of control crop verticillium.
3. verticillium wilt pathogen Antagonistic Fungi as claimed in claim 1 is preparing the application in microbial fertilizer.
4. the application of verticillium wilt pathogen Antagonistic Fungi as claimed in claim 1 in control crop verticillium.
5. verticillium wilt pathogen Antagonistic Fungi as claimed in claim 1 is promoting the application in crop plant growth.
6. for preventing and treating the biocontrol fungicide of crop verticillium, it is characterized in that: it comprises the verticillium wilt pathogen Antagonistic Fungi in claim 1.
7. microbial fertilizer, is characterized in that: it comprises the verticillium wilt pathogen Antagonistic Fungi in claim 1.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106755277A (en) * | 2016-11-14 | 2017-05-31 | 江苏省农业科学院 | A kind of method of the high frequency zone verticillium dahliae antagonistic bacterium from pedotheque |
| CN108901507A (en) * | 2018-07-11 | 2018-11-30 | 新疆农垦科学院 | The method for preventing and treating cotton verticillium wilt |
| CN111718866A (en) * | 2020-05-28 | 2020-09-29 | 南阳师范学院 | Bacillus T6 capable of antagonizing verticillium dahliae and application thereof |
| CN113293108A (en) * | 2021-05-08 | 2021-08-24 | 四川农业大学 | Preparation method of microbial agent for cleaning orchid pot soil, obtained biological agent and application |
| CN114907992A (en) * | 2022-05-18 | 2022-08-16 | 塔里木大学 | Strain C11 for antagonizing phytopathogen and application thereof |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102342300A (en) * | 2011-08-02 | 2012-02-08 | 江苏丘陵地区镇江农业科学研究所 | Composition comprising brevibacillus brevis TW and trichodermin |
| CN102524303A (en) * | 2011-08-31 | 2012-07-04 | 南京农业大学 | Mixture for preventing blight and verticillium wilt of cotton |
| CN103525742A (en) * | 2013-10-30 | 2014-01-22 | 中国农业科学院农产品加工研究所 | Microbial inoculum capable of preventing and treating cotton verticillium wilt and preparation method and application thereof |
-
2016
- 2016-01-28 CN CN201610058155.XA patent/CN105567600A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102342300A (en) * | 2011-08-02 | 2012-02-08 | 江苏丘陵地区镇江农业科学研究所 | Composition comprising brevibacillus brevis TW and trichodermin |
| CN102524303A (en) * | 2011-08-31 | 2012-07-04 | 南京农业大学 | Mixture for preventing blight and verticillium wilt of cotton |
| CN103525742A (en) * | 2013-10-30 | 2014-01-22 | 中国农业科学院农产品加工研究所 | Microbial inoculum capable of preventing and treating cotton verticillium wilt and preparation method and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| 张铎: "《棉花黄萎病拮抗内生细菌的筛选鉴定及其抗菌物质研究》", 《河北师范大学硕士学位论文》 * |
| 郑大玮等: "《农业减灾实用技术手册》", 30 April 2005 * |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106755277A (en) * | 2016-11-14 | 2017-05-31 | 江苏省农业科学院 | A kind of method of the high frequency zone verticillium dahliae antagonistic bacterium from pedotheque |
| CN108901507A (en) * | 2018-07-11 | 2018-11-30 | 新疆农垦科学院 | The method for preventing and treating cotton verticillium wilt |
| CN111718866A (en) * | 2020-05-28 | 2020-09-29 | 南阳师范学院 | Bacillus T6 capable of antagonizing verticillium dahliae and application thereof |
| CN113293108A (en) * | 2021-05-08 | 2021-08-24 | 四川农业大学 | Preparation method of microbial agent for cleaning orchid pot soil, obtained biological agent and application |
| CN114907992A (en) * | 2022-05-18 | 2022-08-16 | 塔里木大学 | Strain C11 for antagonizing phytopathogen and application thereof |
| CN114907992B (en) * | 2022-05-18 | 2023-06-16 | 塔里木大学 | Strain C11 antagonizing plant pathogenic bacteria and application thereof |
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