CN101099492A - Micro-ecological bacterial preparation for killing soil nematodes and its preparation method - Google Patents
Micro-ecological bacterial preparation for killing soil nematodes and its preparation method Download PDFInfo
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- CN101099492A CN101099492A CNA2007100584243A CN200710058424A CN101099492A CN 101099492 A CN101099492 A CN 101099492A CN A2007100584243 A CNA2007100584243 A CN A2007100584243A CN 200710058424 A CN200710058424 A CN 200710058424A CN 101099492 A CN101099492 A CN 101099492A
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- bacillus megaterium
- streptomyces microflavus
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Abstract
The present invention relates to a microecological germicide preparation for killing soil eelworm and its preparation method. Said microecological germicide preparation for killing soil eelworm is made up by using (by wt%) 60%-80%, of bacillus megatherium and 20%-40% of streptolin through a certain preparation process. The soil eelworm-killing rate can be up to 100% and the soil grub-killing rate can be up to above 95%.
Description
Technical field
Biological technical field the present invention relates to a kind of soil nematodes micro-ecological bacterial preparation and preparation method thereof of killing.
Background technology
The soil ecosystem of China or is subjected to heavy damage at present, and soil nematodes is significantly increased, and the kind and the scope of plant disease constantly increase, and this all is the clear proof that soil ecosystem is destroyed.Wherein soil nematodes is the soil pests that directly influences crop yield, the soil of especially planting peanut, Ipomoea batatas, and nematode can be eaten up the peanut in monoblock monoblock soil, Ipomoea batatas or destroy, almost total crop failure.Just sought chemical insecticide in the past, but its 1 year effectively, 1 year nearly unavailable then, and the soil pollution that it causes is all eliminated decades.So it is imperative that the research and development microorganism formulation is killed nematode.More existing researchs both at home and abroad still, yet there are no high density, high activity, the report of the micro-ecological bacterial preparation that the energy large tracts of land is used.
Summary of the invention
Technical problem to be solved by this invention is: a kind of high density, the highly active soil nematodes micro-ecological bacterial preparation and preparation method thereof of killing are provided.
The present invention solves this technical problem the technical scheme that is adopted:
With bacillus megaterium (AS 1.217, Bacillus megaterium) is main, be aided with an amount of streptomyces microflavus (AS4.891, Streptomyces microflavus), and adsorption of immobilization is prepared into bacteria preparation on wheat bran.Two strain bacterial classifications are all purchased the DSMZ in Institute of Microorganism, Academia Sinica.
Killing the soil nematodes micro-ecological bacterial preparation is the complex microorganism composition of above-mentioned bacillus megaterium and streptomyces microflavus optimum organization, and wherein the percentage by weight of each component is: bacillus megaterium 60%～80%, streptomyces microflavus 20%～40%.
The preparation method who kills the soil nematodes micro-ecological bacterial preparation comprises the steps:
1. each slant strains activation, the shake-flask culture respectively that will preserve.Bacillus megaterium and streptomyces microflavus are all used the soluble starch medium culture, medium: pH7.0～7.5,121 ℃ autoclaving 20-30 minute; Bacillus megaterium was cultivated 20～24 hours for 35～40 ℃; Streptomyces microflavus was cultivated 36～50 hours for 28～30 ℃.
2. use the corn starch medium, level liquid enlarged culture bacillus megaterium and streptomyces microflavus; Inoculum concentration 5%～10%.
3. use the corn starch medium, secondary fluid enlargement culture bacillus megaterium and streptomyces microflavus; Inoculum concentration 5%～10%.
4. with wheat bran carrier solid enlarged culture and being immobilized onto on the wheat bran respectively.The culture fluid that adds 10% corn starch before the wheat bran sterilization, solid enlarged culture condition: 30～37 ℃, constantly stir and feed the air of purification.
With bacillus megaterium and streptomyces microflavus through 30～45 ℃ warm air-dry dry, pulverize then.
6. the bacillus megaterium that will be adsorbed on the wheat bran mixes with streptomyces microflavus, and the percentage by weight of each bacterium component is: bacillus megaterium 60%～80%, streptomyces microflavus 20%～40%.
Beneficial effect of the present invention
1) compound bacteria is immobilized onto on the wheat bran, make effective bacterium clump count (CFU) of every gram composite bacteria preparation can be up to 100 hundred million, almost reached most probable number MPN, so far yet there are no the report of high bacteria containing amount bacteria preparation like this. wheat bran can not only provide some nutrient components by being cultivated bacterium, also because its network structure, has bigger specific surface area, so can in unit volume, adhere to the more thalline of adsorption of immobilization.So, effectively bacterium in a single day be manured into soil can very fast formation dominant microflora, the effect of nematode is killed in performance, killing rate is near 100%.Kill or drive away the effect of grub (larva of chafer) in addition, killing rate or expeling rate reach more than 95%.
2) the intestines parietal cell of soil nematodes contains more chitin, and bacillus megaterium can secrete more chitinase.So in a single day soil nematodes runs into bacillus megaterium,, will certainly die at last and die because its intestines wall suffers the destruction of bacillus megaterium chitinase.Bacillus megaterium also has the effect that the phosphorus of soil fixationization is converted into titanium pigment, and titanium pigment is the available phosphorus of plant and microorganism.
Streptomyces microflavus can produce the disease-resistant expelling parasite of antibiotic, and secretion stimulin promotes plant growing, transforms mineral matter.
The two cooperatively interacts, not only desinsection expelling parasite but also promotion, rehabilitating soil ecosystem balance.
Embodiment
Embodiment 1
Follow these steps to carry out:
1) each slant strains with 4 ℃ of preservations activates, respectively shake-flask culture.Bacillus megaterium and streptomyces microflavus are all used the soluble starch medium culture, and it is as follows specifically to fill a prescription: soluble starch 20g, KNO
31g, NaCl 0.5g, K
2HPO
40.5g, MgSO
4.7H
2O 0.5g, FeSO
40.01g, distilled water 1000mL, pH7.2,121 ℃ of autoclavings 20 minutes.Bacillus megaterium was cultivated 24 hours for 37 ℃; Streptomyces microflavus was cultivated 48 hours for 30 ℃.
2) level liquid enlarged culture (condition of culture is the same for bacillus megaterium and the independent enlarged culture of streptomyces microflavus difference, medium corn starch); Inoculum concentration 5%.
3) secondary liquid difference enlarged culture, medium and condition of culture are the same; Inoculum concentration 5%.
4) with wheat bran be carrier solid enlarged culture and being immobilized onto on the wheat bran respectively.Add 10% corn starch culture fluid, solid enlarged culture condition: 30～37 ℃, constantly stir and feed the air of purification before the wheat bran sterilization.
5) 35 ℃ of above-mentioned two kinds of bacterium are warm air-dry dry, moisture content is pulverized below 20% then, and each bacterium that is adsorbed on the wheat bran is mixed by weight percentage: bacillus megaterium 600g, streptomyces microflavus 400g fully mixes, and can obtain bacteria preparation of the present invention.
Embodiment 2
Follow these steps to carry out:
1) each slant strains with 4 ℃ of preservations activates, respectively shake-flask culture.Bacillus megaterium and streptomyces microflavus are all used the soluble starch medium culture, and it is as follows specifically to fill a prescription: soluble starch 20g, KNO
31g, NaCl 0.5g, K
2HPO
40.5g, MgSO
4.7H
2O 0.5g, FeSO
40.01g, distilled water 1000ml, pH7.2,121 ℃ of autoclavings 20 minutes.Bacillus megaterium was cultivated 24 hours for 37 ℃; Streptomyces microflavus was cultivated 48 hours for 30 ℃.
2) level liquid enlarged culture (condition of culture is the same for bacillus megaterium and the independent enlarged culture of streptomyces microflavus difference, medium corn starch); Inoculum concentration 10%.
3) secondary liquid difference enlarged culture, medium and condition of culture are the same; Inoculum concentration 10%.
4) with wheat bran be carrier solid enlarged culture and being immobilized onto on the wheat bran respectively.Add 10% corn starch culture fluid, solid enlarged culture condition: 30～37 ℃, constantly stir and feed the air of purification before the wheat bran sterilization.
5) 45 ℃ of above-mentioned two kinds of bacterium are warm air-dry dry, pulverize then, each bacterium that is adsorbed on the wheat bran is mixed by weight percentage: bacillus megaterium 80kg, streptomyces microflavus 20kg fully mixes, and is packaged in the packaging bag of polyethylene liner, moisture content 20% is valid for one year.
Embodiment 3
Follow these steps to carry out:
1) each slant strains with 4 ℃ of preservations activates, respectively shake-flask culture.Bacillus megaterium and streptomyces microflavus are all used the soluble starch medium culture, and it is as follows specifically to fill a prescription: soluble starch 20g, KNO
31g, NaCl 0.5g, K
2HPO
40.5g, MgSO
4.7H
2O 0.5g, FeSO
40.01g, distilled water 1000ml, pH7.2,121 ℃ of autoclavings 20 minutes.Bacillus megaterium was cultivated 24 hours for 37 ℃; Streptomyces microflavus was cultivated 48 hours for 30 ℃.
2) level liquid enlarged culture (condition of culture is the same for bacillus megaterium and the independent enlarged culture of streptomyces microflavus difference, medium corn starch); Inoculum concentration 10%.
3) secondary liquid difference enlarged culture, medium and condition of culture are the same; Inoculum concentration 5%.
4) with wheat bran be carrier solid enlarged culture and being immobilized onto on the wheat bran respectively.Add 10% corn starch culture fluid, solid enlarged culture condition: 30～37 ℃, constantly stir and feed the air of purification before the wheat bran sterilization.
5) 45 ℃ of above-mentioned two kinds of bacterium are warm air-dry dry, pulverize then, each bacterium that is adsorbed on the wheat bran is mixed by weight percentage: bacillus megaterium 750kg, streptomyces microflavus 250kg fully mixes, and is packaged in the packaging bag of polyethylene liner, moisture content 20% is valid for one year.
Embodiment 4: measure the method for separating soil nematodes:
The centrifugal floatation of the present invention, this method can effectively be separated the most of nematode that comprises line eggs and dead volume, non-motility nematode in the soil.Step is as follows:
Got 20 mesh sieve soil specimen 10g and be placed in the centrifuge tube, added 10～15 milliliters of fully concussions of water, mixed;
With proportion is 10 milliliters of 1.20 aqueous sucrose solutions (sucrose 800g is dissolved in 1 premium on currency), puts the centrifuge tube bottom into pipette or sample injector;
2500g, centrifugal 5min.Solution is divided into two-layer, and nematode accumulates in interlayer, draws with dropper in nematode to the 10 milliliter table ware of interlayer.
To show ware and place counting under the anatomical lens.
The advantage of this method: only need be once centrifugal, the nematode water liquid of collection is clear, distinguishes the form of nematode easily.
Points for attention: 1) soil specimen tends to be mixed with canebreak, in order to reduce the mixed of nematode is stirred, be preferably in the sampling before soil specimen is crossed 20 mesh sieves;
2) because of the relation of the sucrose liquid that adds what and centrifugal speed, interlayer is very not clear sometimes, when running into this situation, can suitably get a little aqueous solution, in order to avoid lose nematode more.
Claims (2)
1. micro-ecological bacterial preparation of killing soil nematodes, it is characterized in that: this bacteria preparation is made up of the complex microorganism of bacillus megaterium (Bacillusmegaterium) and streptomyces microflavus (Staestomyces microflavus) optimum organization, wherein the percentage by weight of each bacterium component is: bacillus megaterium 60%～80%, streptomyces microflavus 20%～40%.
2. the preparation method who kills the micro-ecological bacterial preparation of soil nematodes as claimed in claim 1 is characterized in that it comprises the steps:
1) each slant strains activation, the shake-flask culture respectively that will preserve; Bacillus megaterium and streptomyces microflavus are all used the soluble starch medium culture, medium: pH7.0～7.5,115～121 ℃ autoclaving 20～30 minutes; Bacillus megaterium was cultivated 20～24 hours for 35～40 ℃; Streptomyces microflavus was cultivated 36～50 hours for 28～30 ℃;
2) the level liquid enlarged culture of huge gemma bar and streptomyces microflavus (20L jar); Inoculum concentration 5%～10%;
3) the secondary fluid enlargement culture of bacillus megaterium and streptomyces microflavus (200L jar); Inoculum concentration 5%～10%;
4) be that carrier carries out the solid enlarged culture respectively and is immobilized onto on the wheat bran with wheat bran.Solid enlarged culture condition: 30～37 ℃, constantly stir and feed the air of purification;
5) with bacillus megaterium and streptomyces microflavus respectively 30～45 ℃ warm air-dry dry, pulverize then;
6) dry bacillus megaterium is mixed with streptomyces microflavus, the percentage by weight of each bacterium component is: bacillus megaterium 60%～80%, streptomyces microflavus 20%～40%.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100584243A CN101099492A (en) | 2007-07-30 | 2007-07-30 | Micro-ecological bacterial preparation for killing soil nematodes and its preparation method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2007100584243A CN101099492A (en) | 2007-07-30 | 2007-07-30 | Micro-ecological bacterial preparation for killing soil nematodes and its preparation method |
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| CN101099492A true CN101099492A (en) | 2008-01-09 |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101822273A (en) * | 2010-04-21 | 2010-09-08 | 中国农业科学院草原研究所 | Application of streptomyces microflavus in alfalfa disease control |
| CN101508968B (en) * | 2009-03-06 | 2011-04-20 | 中国农业大学 | Biological medicament for preventing and treating plant parasitic nematode and uses thereof |
| CN102690151A (en) * | 2012-06-28 | 2012-09-26 | 赵鹏 | Composite microbial inoculum organic multielement compound fertilizer and production method thereof |
| CN107217049A (en) * | 2017-06-29 | 2017-09-29 | 烟台固特丽生物科技股份有限公司 | One plant growth regulators are applied to method prepared by drappus microbial inoculum |
| CN114395548A (en) * | 2021-12-31 | 2022-04-26 | 春华秋实科技集团有限公司 | Compound microbial preparation for preventing and treating root-knot nematode and application thereof |
-
2007
- 2007-07-30 CN CNA2007100584243A patent/CN101099492A/en active Pending
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101508968B (en) * | 2009-03-06 | 2011-04-20 | 中国农业大学 | Biological medicament for preventing and treating plant parasitic nematode and uses thereof |
| CN101822273A (en) * | 2010-04-21 | 2010-09-08 | 中国农业科学院草原研究所 | Application of streptomyces microflavus in alfalfa disease control |
| CN102690151A (en) * | 2012-06-28 | 2012-09-26 | 赵鹏 | Composite microbial inoculum organic multielement compound fertilizer and production method thereof |
| CN102690151B (en) * | 2012-06-28 | 2014-03-05 | 赵鹏 | Composite microbial inoculum organic multielement compound fertilizer and production method thereof |
| CN107217049A (en) * | 2017-06-29 | 2017-09-29 | 烟台固特丽生物科技股份有限公司 | One plant growth regulators are applied to method prepared by drappus microbial inoculum |
| CN114395548A (en) * | 2021-12-31 | 2022-04-26 | 春华秋实科技集团有限公司 | Compound microbial preparation for preventing and treating root-knot nematode and application thereof |
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