CN100513554C - Short small bacillus TW and viable bacteria preparation for rice blast prevention and cure - Google Patents
Short small bacillus TW and viable bacteria preparation for rice blast prevention and cure Download PDFInfo
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- CN100513554C CN100513554C CNB2007100215094A CN200710021509A CN100513554C CN 100513554 C CN100513554 C CN 100513554C CN B2007100215094 A CNB2007100215094 A CN B2007100215094A CN 200710021509 A CN200710021509 A CN 200710021509A CN 100513554 C CN100513554 C CN 100513554C
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- rice blast
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- bacillus pumilus
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Abstract
The invention discloses a short small germ bacillus TW and viable bacteria preparation with this bacterium to prevent and cure rice blast, which is characterized by the following: possessing preserved number at CCTCC NO: M207014 of the short small germ bacillus TW; incorporating short small germ bacillus 1. 0X1010cfu/g-2. 0X 1010cfu/g in solid preparation composition of this viable bacteria preparation; including additive; setting the composition and mass percent of the additive as 0. 05%-0. 10% ultraviolet screener, 0. 5%-1. 5% humectant, 1. 0%-5. 0% preservative, 0. 5%-1. 5% conservative and solid filler. This bacillus TW possesses strong antagonistic action for rice blast.
Description
Technical field
The present invention relates to a kind of short bacillus pumilus TW and with the active bacteria formulation of the control rice blast of this strain preparation.
Background technology
Rice blast (Blast Fungus) is one of the most serious rice disease of rice district, whole world harm.This disease all can work the mischief to paddy rice throughout the year, and its harm spreads all over each position of paddy rice, and seedling rice blast, leaf pest, pulvinus pest, joint rice blast, panicle blast, branch stalk pest and grain pest etc. are arranged.Generally cause the paddy rice underproduction 10%～20%.Area is taking place China's year all more than 3,800,000 hm2 in this disease, and year loss paddy reaches several hundred million kg.At present, the utilization of the anti-pest kind of paddy rice and the use of chemical pesticide are still the effective measure of control rice blast, but because of the simplification of anti-pest kind, the complicacy of physiological races of rice blast fungus heredity and pathogenic diversity, and the resistance of the toxicity of chemical pesticide and pathogenic bacteria, anti-pest kind of paddy rice and chemical pesticide use on producing and all are subjected to certain restriction.For this reason, the biological control of rice blast particularly the research and development of biological pesticide be the needs of Rice Production Sustainable development.Microbial pesticide is less to the selective pressure of environment, and is difficult for making the target pathogenic bacteria to develop immunity to drugs, and therefore, the research of biogenic pesticide will provide the safer approach of preventing and treating rice blast more efficiently for people.
In recent years, except safety non-toxic, going back response environment has less selective pressure to the requirement of agricultural chemicals.Biological pesticide best embodies the characteristics of above-mentioned control, its action target or generally all be polyfactorial to the effect of pathogenic bacteria, or claim multi-purpose, so be difficult for making the target bacterium to develop immunity to drugs.Under the prerequisite that guarantees rice yield, the use of biological pesticide is indispensable.At present, rice blast biological control means are had: (1) is administration of antibiotics directly, as kasugamycin, kasugamycin, miewensu-S, Blasticidin, qingfengmeisu (its streptomycete, Streptomyces kasugensis Umezawa, grey produce look streptomycete, GriseochromogenesFukunaga, the rich streptomycete of celebrating, actinomycetes) microbe-derived springtime; (2) biocontrol bacteria and fungi preparation are as genus bacillus (Bacillussubtilis), Bacillus amyloliquef aciens RC-2, bacillus cereus, pseudomonas (Pseudomonas aeruginosa PR3 (NRRL B-18602) and fungus Trichoderma.But up to the present, the report and the related application of short bacillus pumilus still of no use (Brevibacillus brevis) control rice blast.
Summary of the invention
Technical problem to be solved by this invention is to disclose a kind of short bacillus pumilus TW, and a kind of active bacteria formulation that comprises short bacillus pumilus TW and rice blast is had the control rice blast of better prevention effect is provided.
Microorganism of the present invention is that screening obtains in a kind of endophytic bacteria out of office has the short bacillus pumilus TW of strong antagonistic action to rice blast (the Latin generic name is: Brevibacillus brevis), this bacterial classification on February 9th, 2007 in China's typical culture collection center preservation, preserving number is: CCTCC NO:M207014.
Above-mentioned bacterial classification has following feature:
1, morphological specificity: bacterium is direct rod shape, tool nose circle or square end, and colonial morphology is round, protuberance, the edge is smooth, and yellow-white is to the oyster white bacterium colony;
2, physiological and biochemical property: Gram-positive, aerobic, peritrichous can produce gemma, gemma ellipse or column;
3, fermented liquid can produce multiple antibacterial peptide;
4, fermented liquid energy antagonism rice blast fungus, Rhizoctonia solani Kuhn, the sick fungi of various plants such as rhizoctonia cerealis and Sclerotinia sclerotiorum.
A kind of form of the active bacteria formulation of control rice blast of the present invention is a solid preparation, comprises short bacillus pumilus bacterial classification 1.0 * 10 in its composition
10Cfu/g～2.0 * 10
10Cfu/g, the additive that also includes, the composition of additive and weight ratio are: anti-ultraviolet radiation agent 0.05%～0.10%, wetting agent 0.5%～1.5%, protective material 1.0%～5.0%, sanitas 0.5%-1.5%, all the other are solid filler.
The another kind of form of the active bacteria formulation of control rice blast of the present invention is a liquid preparation, comprises short bacillus pumilus bacterial classification 1.0 * 10 in its composition
10Cfu/ml～2.0 * 10
10Cfu/ml also includes additive, and the composition of additive and weightmeasurement ratio are: sanitas 0.5%～1.5%, and frostproofer 0.5%～1.5%, thickness permeability-reducing admixture 5.0%～10.0%, wetting agent 0.5%～1.5%, all the other are damping fluid.
Its advantage of the present invention and positively effect are as follows:
1, indoor antagonistic activity measurement result shows, the short bacillus pumilus TW of the present invention has extremely strong bacteriostatic activity, EC to the Pyricularia oryzae mycelial growth
50Value is that (95% fiducial limit is 0.2801～20.3491mg/L) to 6.1008mg/L, and has the EC of the contrast medicament Bacilluscereus, of broad spectrum antagonistic activity
50Value is that (95% fiducial limit is 65.8516～129.7483mg/L) to 92.6835mg/L.Two kinds of medicament EC
5095% fiducial limit of value is not overlapping, illustrates that the medicament differences is remarkable;
2, the anti-effect of the field demonstration of active bacteria formulation of the present invention shows that this microbial inoculum reaches more than 83% the rice blast prevention effect;
3, because active bacteria formulation of the present invention is a biotechnological formulation, there are not a series of environment and safety problem that use brought because of chemical pesticide, thereby the non-environmental pollution control of this pernicious disease of assurance rice blast, the while can reduce the consumption of other chemical pesticides, for the peasant reduces expenses.
Embodiment
Short bacillus pumilus TW of the present invention (Brevibacillus brevis) bacterial strain is exclusively preserved by the Zhenjiang City,Agriculture Science Research Institute.
The active bacteria formulation of control rice blast of the present invention can adopt liquid preparation and solid preparation, and the specific embodiment of its preparation process is as follows:
Embodiment one, and the preparation of liquid preparation may further comprise the steps:
(1) cultivation of first order seed nutrient solution
One of picking grows in the single bacterium colony spot of short bacillus pumilus TW on the R2A substratum, in the 250ml triangular flask that 50mlR2A nutrient solution (being not add agar in the R2A substratum) is housed, puts 30 ℃, shaking culture 16h on the 200rpm shaking table.
(2) cultivation of secondary seed nutrient solution
The first order seed nutrient solution is inoculated in the R2A nutrient solution by the 1:50 amplification, puts the same terms and cultivate 24h.
(3) go up a jar fermentation
The secondary seed nutrient solution is inoculated in fermentation culture (composition: glucose 1.0%, soybean cake powder 2.50%, peptone 0.4%, NaCl 0.2%, (NH by the 1:200 amplification
4)
2SO
40.6%, CaCO
30.4%, MgSO
40.4%).Fermentation culture conditions is: pH7.2, dissolved oxygen are 100%, and mixing speed is 350rmp, and 30 ℃, 24h goes out ferment.
(4) centrifugal collection thalline is with the 0.01M phosphoric acid buffer thalline that suspends again.
(5) liquid preparation obtains: with the above-mentioned furnishing of suspension again 1.0 * 10
10Behind the cfu/ml, add additive by a certain percentage, i.e. 0.1% (W/V) rhodamine B, 1.0% (W/V) Sodium Benzoate, 0.5% propylene glycol (W/V) and 5.0% (W/V) xanthan gum, sodium lignosulfonate 0.5%～1.5% (W/V), complement to 100% with the 0.01M phosphoric acid buffer, the aseptic bottle packing.
Embodiment two; the preparation process of solid preparation comprise cultivation, the secondary seed nutrient solution of the first order seed nutrient solution identical with embodiment one cultivation, go up a jar fermentation, centrifugal collection thalline step, add in the suspension after centrifugal protective material sucrose 1.0% postlyophilization (or 60 ℃ of dryings) become solid after comminution by gas stream get female bacterium powder.Add additive in proportion, i.e. 0.1% (W/W) rhodamine B, 1.0% (W/W) Sodium Benzoate and 1.0% sodium lignosulfonate (W/W), regulating bacteria concentration with potter's clay is 1.0 * 10
10Cfu/g, the sterile bag packing.
The land for growing field crops method of application of active bacteria formulation of the present invention is: 800-1000 times of diluent, thin spraying.
Implementation result of the present invention such as next:
(1) the laboratory experiment result is shown in table 1, table 2.Laboratory experiment shows, short bacillus pumilus TW has extremely strong bacteriostatic activity to the Pyricularia oryzae mycelial growth, the EC50 value is 6.1008mg/L (table 2), 95% fiducial limit is 0.2801～20.3491mg/L, and the EC50 value with contrast medicament Bacilluscereus, of broad spectrum antagonistic activity is 92.6835mg/L, and 95% fiducial limit is 65.8516～129.7483mg/L.95% fiducial limit of two kinds of medicament EC50 values is not overlapping, illustrates that the medicament differences is remarkable.
The short bacillus pumilus TW of table 1 measures (original recorded data) to Pyricularia oryzae mycelial growth bacteriostasis rate
The statistics that the short bacillus pumilus TW of table 2 measures the indoor bacteriostatic activity of Pyricularia oryzae
| The medicament title | Toxicity regression line Y= | EC 50Value (mg/L) | 95% fiducial limit | r | Relative virus force index |
| Short bacillus pumilus TW | 6.2308+1.0133X | 6.1008 | 0.2801～ 20.3491 | 0.9816 | 15.2 |
| Bacilluscereus, | 5.0445+1.3487X | 92.6835 | 65.8516～ 129.7483 | 0.9712 | 1.0 |
(2) control in field demonstration test result is as shown in table 3.Test-results shows that this bacteria preparation solid preparation is to the preventive effect of rice panicle blast, and the preventive effect on two kinds of rice varieties is all more than 80%; Liquid preparation on two kinds of rice varieties preventive effect all more than 90%.Preventive effect difference between liquid preparation and chemical agent (tricyclazole) is not remarkable.
The demonstration of white farm prevents and treats the result behind table 3 paddy rice panicle blast in 2006 Jurong
| Formulation | Kind | Demonstration area (mu) | Strain sickness rate (%) | Preventive effect (%) |
| TW1 (liquor) | No. 13, military fortune chaff | 71 | 0.10 | 98.4 |
| TW2 (liquor) | 7319 | 5 | 0.25 | 96.0 |
| TW3 (pulvis) | No. 13, military fortune chaff | 35 | 0.64 | 89.7 |
| TW5 (pulvis) | No. 7, military fortune chaff | 19 | 1.00 | 83.9 |
| Tricyclazole (chemical agent contrast) | No. 13, military fortune chaff | 55 | 0.43 | 93.1 |
| Blank | No. 7, military fortune chaff | 3 | 6.20 | 0 |
Annotate: the control time is the paddy rice cut phase, and control once.TW: refer to short bacillus pumilus TW
Claims (5)
1, (the Latin generic name is: Brevibacillus brevis), preserving number is CCTCC NO:M207014 to a kind of short bacillus pumilus TW.
2, a kind of active bacteria formulation of preventing and treating rice blast is characterized in that: it is a solid preparation, comprises short bacillus pumilus t bacteria W1.0 as claimed in claim 1 * 10 in its composition
10Cfu/g～2.0 * 10
10Cfu/g also includes additive, and the composition of additive and weight ratio are: anti-ultraviolet radiation agent 0.05%～0.10%, and wetting agent 0.5%～1.5%, protective material 1.0%～5.0%, sanitas 0.5%-1.5%, all the other are solid filler.
3, the active bacteria formulation of control rice blast according to claim 2 is characterized in that: described anti-ultraviolet radiation agent is a rhodamine B, and described wetting agent is a sodium lignosulfonate, and described sanitas is a sodium benzoate, and described protective material is a sucrose.
4, a kind of active bacteria formulation of preventing and treating rice blast is characterized in that: it is a liquid preparation, comprises short bacillus pumilus t bacteria W 1.0 * 10 as claimed in claim 1 in its composition
10Cfu/ml～2.0 * 10
10Cfu/ml also includes additive, and the composition of additive and weightmeasurement ratio are: sanitas 0.5%～1.5%, and frostproofer 0.5%～1.5%, thickness permeability-reducing admixture 5.0%～10.0%, wetting agent 0.5%～1.5%, all the other are damping fluid.
5, the active bacteria formulation of control rice blast according to claim 4 is characterized in that: described sanitas is a sodium benzoate, and described frostproofer is a propylene glycol, and described thickness permeability-reducing admixture is an xanthan gum, and described wetting agent is a sodium lignosulfonate.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CNB2007100215094A CN100513554C (en) | 2007-04-19 | 2007-04-19 | Short small bacillus TW and viable bacteria preparation for rice blast prevention and cure |
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|---|---|---|---|
| CNB2007100215094A CN100513554C (en) | 2007-04-19 | 2007-04-19 | Short small bacillus TW and viable bacteria preparation for rice blast prevention and cure |
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| CN100513554C true CN100513554C (en) | 2009-07-15 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103563991A (en) * | 2012-07-31 | 2014-02-12 | 江苏丘陵地区镇江农业科学研究所 | Preparation method of brevibacillus brevis water dispersible granules |
| CN103563992A (en) * | 2012-07-31 | 2014-02-12 | 江苏丘陵地区镇江农业科学研究所 | Combined preparation of biological pesticide and chemical pesticide for preventing and treating rice blast |
Families Citing this family (6)
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| CN101928680B (en) * | 2009-11-19 | 2012-11-14 | 无锡亚克生物科技有限公司 | Bacillus simplex RJGP41 and application thereof |
| CN101942403B (en) * | 2010-06-12 | 2012-06-06 | 浙江大学 | Bacillus pumilus as well as culture method and application thereof |
| CN102125059B (en) * | 2010-12-03 | 2013-03-06 | 江苏苏滨生物农化有限公司 | Brevibacillus brevis TW effervescent granule and preparation method thereof |
| 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 |
| CN105104048B (en) * | 2015-09-28 | 2018-05-18 | 江苏丘陵地区镇江农业科学研究所 | It is a kind of to induce identification, the method for screening rice anti-rice blast breeding material |
| CN110396490B (en) * | 2019-08-15 | 2020-10-09 | 江苏苏滨生物农化有限公司 | Method for culturing brevibacillus brevis |
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- 2007-04-19 CN CNB2007100215094A patent/CN100513554C/en not_active Expired - Fee Related
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| Mode of antagonism of Brevibacillus brevis againstBotrytiscinerea in vitro. S.G. Edwards.Journal of Applied Microbiology,Vol.91 . 2001 |
| Mode of antagonism of Brevibacillus brevis againstBotrytiscinerea in vitro. S.G. Edwards.Journal of Applied Microbiology,Vol.91 . 2001 * |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103563991A (en) * | 2012-07-31 | 2014-02-12 | 江苏丘陵地区镇江农业科学研究所 | Preparation method of brevibacillus brevis water dispersible granules |
| CN103563992A (en) * | 2012-07-31 | 2014-02-12 | 江苏丘陵地区镇江农业科学研究所 | Combined preparation of biological pesticide and chemical pesticide for preventing and treating rice blast |
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| CN101063094A (en) | 2007-10-31 |
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Application publication date: 20071031 Assignee: Jiangsu Subin Agrochemical Co., Ltd. Assignor: Zhenjiang Institute of Agricultural Sciences, Jiangsu Hilly Region Contract record no.: 2010320000405 Denomination of invention: Short small bacillus TW and viable bacteria preparation for rice blast prevention and cure Granted publication date: 20090715 License type: Exclusive License Record date: 20100416 |
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Granted publication date: 20090715 Termination date: 20170419 |