CN101759491A - Bacillus subtilis suspended seed coating agent, preparation method and application thereof - Google Patents
Bacillus subtilis suspended seed coating agent, preparation method and application thereof Download PDFInfo
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- CN101759491A CN101759491A CN200910223355A CN200910223355A CN101759491A CN 101759491 A CN101759491 A CN 101759491A CN 200910223355 A CN200910223355 A CN 200910223355A CN 200910223355 A CN200910223355 A CN 200910223355A CN 101759491 A CN101759491 A CN 101759491A
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Abstract
The invention discloses a bacillus subtilis aqueous-suspension type biological seed coating agent, a preparation method and application thereof. The preparation method mainly takes bacillus subtilis as an effective component and adds hybrid film-forming agents, nutritional agents, thickeners, antifreeze, preservatives, coloring agents and water so as to obtain a bacillus subtilis aqueous-suspension seed coating agent which is free from environment pollution, low in toxicity, safe and long in storage life through simple making processes. The seed coating agent not only has the function of promoting plant growth, but also can effectively prevent and control soil-borne diseases of plants and increase plant production, and particularly has significant effect of preventing and controlling soybean root rot, soybean seedling blight and other soybean soil-borne diseases.
Description
Technical field
The present invention relates to a kind of biological pesticide preparation, specifically, is a kind of subtilis aqueous suspension kind clothing agent and its production and use.
Background technology
Plant soil-borne diseases is the important Plant diseases of a class, and its incidence is the trend that rises year by year, has caused heavy losses to rural economy.Aspect the preventing and treating of soil-borne disease, plant the clothing agent and occupy critical role always.Planting the clothing agent is that a class is used for the pesticide formulation that seed pelleting is handled, and is the fertile compound seed pelleting product of mainly being processed into by sterilant, sterilant, compound manure, trace element, plant-growth regulator, sustained release dosage and membrane-forming agent etc. of medicine.Planting the clothing agent is carrier with the seed; be attached on the seed by membrane-forming agent; difficult drop-off; after planting again seed is formed a protection barrier; swelling and not dissolving at once after absorbing water; effective constituent can be absorbed by plant root gradually, the pathogenic in the soil and subterranean pest-insect is played the effect of disease-prevention and insecticide.
For a long time, the market of planting the clothing agent is based on the agent of chemical seed clothing, but the high toxicity of chemical pesticide makes kind of clothing agent all can produce certain toxic action in the process of processing, storage, application, also can cause adverse consequencess such as pesticide residue, environmental pollution, pathogenic bacteria develop immunity to drugs simultaneously.The agent of biotype kind clothing has low toxicity even nontoxic character, can fundamentally overcome the above-mentioned disadvantage of chemical seed clothing agent, thereby people are all over the world exploring how to make the agent of biotype kind clothing with useful microbial preparation replacement chemical pesticide.
Subtilis (Bacillus subtilis) is that (Plant Growth-Promoting Rhizobacteria PGPR), has a liking for temperature, aerobic, product gemma to the short living bacterium of a kind of important plant rhizosphere, and physiological property is various; Widely distributed at occurring in nature, and separation and Culture very easily, can produce multiple antibiotic and enzyme, have broad spectrum antibiotic activity and extremely strong anti-adversity ability.The bacillus subtilis bacteria growing is fast, nutrition is simple, heat-resisting, to people and animals' toxicological harmless, outstanding characteristic such as free from environmental pollution, make it can survive at multiple physical environment and under artificial environments such as production, formulation processing, grow surely and breeding, be a kind of ideal biocontrol bacteria, but do not see with the subtilis to be the report that main active ingredient is made biological seed coating agent at present.
Summary of the invention
The object of the invention is to provide the subtilis aqueous suspension kind clothing agent of a kind of, low toxicity nuisanceless to environment, safety, and it not only has growth-promoting functions to plant, and can effectively prevent and treat plant soil-borne diseases, improves plant biomass; Another object of the present invention is to provide the preparation method of above-mentioned subtilis aqueous suspension kind clothing agent.
Above-mentioned purpose realizes by following technique means:
The agent of a kind of subtilis aqueous suspension kind clothing, mixed by following composition according to weight percent: subtilis aqueous suspensions 10%～30%, be mixed into film 60%～80%, nutrition agent 2.5%～5%, thickening material 1%～5%, frostproofer 3%～8%, sanitas 0.1%～0.5%, tinting material 0.5%～1.5%, all the other are water.
Described subtilis aqueous suspensions is the main active ingredient that the present invention plants the clothing agent, prepare through following steps: earlier with subtilis through liquid submerged fermentation (adopting conventional zymotechnique), centrifugally then remove supernatant, at last with the suitable quantity of water thalline that suspends, make cell concentration in the aqueous suspensions 〉=10
8Individual/mL.
Preferably, described subtilis is screened subtilis 69 bacterial strains that get from the natural plant rhizosphere soil for researchist of the present invention, this bacterium all has good bacteriostasis property for various plants pathogenetic bacteria, fungi, and is particularly particularly remarkable for the fungistatic effect of soybean soil-borne disease pathogenic bacterias such as pathogen of soybean root rot, soybean rhizoctonia solani.
The described film that is mixed into is to be that to be 4% chitosan solution mix with 8: 2 weight ratio for 5% polyvinyl alcohol water solution and weightmeasurement ratio by weightmeasurement ratio.Described weightmeasurement ratio is that 4% chitosan solution is to be dissolved with 4% weightmeasurement ratio by chitosan to get in 2% aqueous acetic acid.
Described nutrition agent is one or more the mixture in vitriol, nitrate, phosphoric acid salt, the silicate.
Described thickening material is one or more mixtures in xanthan gum, carboxymethyl cellulose, the magnesium aluminum silicate.
Described frostproofer is an ethylene glycol.
Described sanitas is one or more mixtures in Sodium Benzoate, sodium sorbate, the Paraformaldehyde 96.
Described tinting material is sarranine or basic rhodamine.
The preparation method of above-mentioned subtilis aqueous suspension kind clothing agent comprises the steps:
(1) with subtilis earlier through liquid submerged fermentation (adopting conventional zymotechnique), centrifugally then remove supernatant, (make cell concentration in the aqueous suspensions 〉=10 with the suitable quantity of water thalline that suspends again
8Individual/mL), it is standby to get the subtilis aqueous suspensions;
(2) polyvinyl alcohol is dissolved in 90 ℃ of hot water with 5% weightmeasurement ratio, chitosan is dissolved in 2% aqueous acetic acid with 4% weightmeasurement ratio, according to 8: 2 weight ratio the two is mixed then, make the required film that is mixed into;
(3) in being mixed into film, add nutrition agent, thickening material, frostproofer, sanitas, tinting material, stir, to the subtilis aqueous suspensions of wherein sneaking into step (1), add water more then, stir and promptly get bacillus subtilis suspended seed coating agent of the present invention.
Bacillus subtilis suspended seed coating agent of the present invention and soybean or other plant seed carry out dressing with 1: 50 weight ratio, can effectively prevent and treat the multiple soil-borne disease of soybean or other plant.
Low production cost, the manufacture craft of bacillus subtilis suspended seed coating agent of the present invention are simple, use very conveniently, and the shelf lives is long; Wrap on the seed and can effectively prevent and treat the various plants soil-borne disease, particularly particularly remarkable for the preventive and therapeutic effect of soybean soil-borne diseases such as root rot, soybean damping-off.
Description of drawings
Fig. 1 is that subtilis 69 suspension seed-coating agents of embodiment 9 are handled the soybean seeds growth figure after 30 days;
Fig. 2 is that the Lip river bacterium nitrile of embodiment 9 is handled the soybean seeds growth figure after 30 days;
Fig. 3 is that the clear water of embodiment 9 is handled the soybean seeds growth figure after 30 days.
Embodiment
Below further describe the present invention by embodiment, as known by the technical knowledge, the present invention also can describe by other the scheme that does not break away from the technology of the present invention feature, thus all within the scope of the present invention or the change that is equal in the scope of the invention all be included in the invention.
Embodiment 1: preparation subtilis 69 aqueous suspensions
With subtilis earlier through liquid submerged fermentation (adopting conventional zymotechnique), centrifugally then remove supernatant, (make cell concentration in the aqueous suspensions 〉=10 with the suitable quantity of water thalline that suspends again
8Individual/mL), promptly.
Embodiment 2: preparation is mixed into film
Polyvinyl alcohol is dissolved in 90 ℃ of hot water with 5% weightmeasurement ratio, chitosan is dissolved in 2% aqueous acetic acid with 4% weightmeasurement ratio, according to 8: 2 weight ratio the two is mixed then, promptly.
Embodiment 3: preparation subtilis 69 suspension seed-coating agents
(1) according to following composition and weight percent batching: subtilis 69 aqueous suspensions 10% of embodiment 1, embodiment 2 is mixed into film 80%, xanthan gum 1%, ammonium sulfate 2.5%, ethylene glycol 3%, Sodium Benzoate 0.1%, sarranine 1%, water 2.4%;
(2) in being mixed into film, add xanthan gum, ammonium sulfate, ethylene glycol, Sodium Benzoate, sarranine, stir, to wherein sneaking into subtilis 69 aqueous suspensions, add water more then, stir and promptly get subtilis 69 suspension seed-coating agents.
Embodiment 4: preparation subtilis 69 suspension seed-coating agents
(1) according to following composition and weight percent batching: subtilis 69 aqueous suspensions 15% of embodiment 1, embodiment 2 is mixed into film 60%, xanthan gum 5%, nutrition agent (potassium primary phosphate and ammonium nitrate form according to 1: 1 mixed in molar ratio) 3.0%, ethylene glycol 5%, Sodium Benzoate 0.3%, sarranine 0.5%, water 11.2%;
(2) in being mixed into film, add xanthan gum, nutrition agent, ethylene glycol, Sodium Benzoate, sarranine, stir, to wherein sneaking into subtilis 69 aqueous suspensions, add water more then, stir and promptly get subtilis 69 suspension seed-coating agents.
Embodiment 5: preparation subtilis 69 suspension seed-coating agents
(1) according to following composition and weight percent batching: subtilis 69 aqueous suspensions 15% of embodiment 1, embodiment 2 is mixed into film 65%, xanthan gum 3.5%, sal epsom 5%, ethylene glycol 3%, Paraformaldehyde 96 0.25%, basic rhodamine 0.5%, water 7.75%;
(2) in being mixed into film, add xanthan gum, sal epsom, ethylene glycol, Paraformaldehyde 96, basic rhodamine, stir, to wherein sneaking into subtilis 69 aqueous suspensions, add water more then, stir and promptly get subtilis 69 suspension seed-coating agents.
Embodiment 6: preparation subtilis 69 suspension seed-coating agents
(1) according to following composition and weight percent batching: subtilis 69 aqueous suspensions 20% of embodiment 1, embodiment 2 is mixed into film 70%, carboxymethyl cellulose 1.5%, zinc sulfate 2.5%, ethylene glycol 5%, sodium sorbate 0.5%, basic rhodamine 0.5%;
(2) in being mixed into film, add carboxymethyl cellulose, zinc sulfate, ethylene glycol, sodium sorbate, basic rhodamine, stir, to wherein sneaking into subtilis 69 aqueous suspensions, add water more then, stir and promptly get subtilis 69 suspension seed-coating agents.
Embodiment 7: preparation subtilis 69 suspension seed-coating agents
(1) according to following composition and weight percent batching: subtilis 69 aqueous suspensions 10% of embodiment 1, embodiment 2 is mixed into film 75%, magnesium aluminum silicate 1%, zinc sulfate 2.5%, ethylene glycol 4%, Sodium Benzoate 0.2%, sarranine 1%, water 6.3%;
(2) in being mixed into film, add magnesium aluminum silicate, zinc sulfate, ethylene glycol, Sodium Benzoate, sarranine, stir, to wherein sneaking into subtilis 69 aqueous suspensions, add water more then, stir and promptly get subtilis 69 suspension seed-coating agents.
Embodiment 8: preparation subtilis 69 suspension seed-coating agents
(1) according to following composition and weight percent batching: subtilis 69 aqueous suspensions 20% of embodiment 1, embodiment 2 is mixed into film 65%, xanthan gum 1%, nutrition agent (zinc sulfate and potassium primary phosphate form according to 1: 1 mixed in molar ratio) 2.5%, ethylene glycol 5%, Sodium Benzoate 0.2%, sarranine 1%, water 5.3%;
(2) in being mixed into film, add xanthan gum, nutrition agent, ethylene glycol, Sodium Benzoate, sarranine, stir, to wherein sneaking into subtilis 69 aqueous suspensions, add water more then, stir and promptly get subtilis 69 suspension seed-coating agents.
Embodiment 9:
Subtilis 69 suspension seed-coating agents that embodiment 3 is made carry out dressing with 1: 50 weight ratio to soybean, establish two groups of contrasts simultaneously, and the first is carried out dressing with 1: 150 weight ratio to soybean seeds with chemical seed clothing agent Lip river bacterium nitrile; It two is to handle soybean seeds with clear water.Three kinds of soybean seeds of handling with different modes are respectively dried, sow in a conventional manner then and filling sterilization soil and admixing in the flowerpot of soybean sickle spore bacterium.After 30 days, observe through different modes and handle the situation of growing and the incidence of soybean afterwards, result such as table 1 and Fig. 1～shown in Figure 3:
Table 1:
| Handle | Percentage of germination (%) | Average plant height (cm) | Average fresh weight (g) | Disease index (%) | Preventive effect (%) |
| Subtilis 69 aqueous suspension kind clothing agent | ??88 | ??50.39 | ??5.63 | ??21.68 | ??73.31 |
| Lip river bacterium nitrile | ??85 | ??35.64 | ??3.44 | ??31.52 | ??61.20 |
| Clear water (CK) | ??70 | ??28.52 | ??2.90 | ??81.24 | ??-- |
By table 1 and Fig. 1～Fig. 3 as can be known, subtilis 69 suspension seed-coating agents of the present invention can effectively promote soybeans they grow, improve seedling rate, improve the soybean resistance against diseases, good to the root rot prevention effect, show better disease prevention growth-promoting effect with respect to commercially available agricultural chemicals Lip river bacterium nitrile.
Embodiment 10:
Subtilis 69 suspension seed-coating agents that embodiment 5 is made carry out dressing with 1: 50 weight ratio to soybean, and the soybean seeds of handling with clear water in contrast.Two kinds of soybean seeds of handling with different modes are respectively dried, sow in a conventional manner then and filling sterilization soil and admixing in the flowerpot of soybean rhizoctonia solani.After 30 days, observe through different modes and handle the situation of growing and the incidence of soybean afterwards, the result is as shown in table 2:
Table 2:
| Handle | Percentage of germination (%) | Average plant height (cm) | Average fresh weight (g) | Disease index (%) | Preventive effect (%) |
| Subtilis 69 aqueous suspension kind clothing agent | ??86 | ??52.26 | ??6.01 | ??21.31 | ??75.02 |
| Clear water (CK) | ??71 | ??29.47 | ??3.22 | ??85.31 | ??-- |
As shown in Table 2, subtilis 69 suspension seed-coating agents of the present invention can effectively promote soybeans they grow, improve seedling rate, improve the soybean resistance against diseases, and especially the prevention effect to the soybean damping-off is good.
Claims (6)
1. subtilis aqueous suspension type kind clothing agent, it is characterized in that, mixed by following composition according to weight percent: subtilis aqueous suspensions 10%～30%, be mixed into film 60%～80%, nutrition agent 2.5%～5%, thickening material 1%～5%, frostproofer 3%～8%, sanitas 0.1%～0.5%, tinting material 0.5%～1.5%, all the other are water;
Cell concentration 〉=10 in the described subtilis aqueous suspensions
8Individual/mL;
The described film that is mixed into is to be that to be 4% chitosan solution mix with 8: 2 weight ratio for 5% polyvinyl alcohol water solution and weightmeasurement ratio by weightmeasurement ratio; Described weightmeasurement ratio is that to be chitosan with 4% weightmeasurement ratio dissolve in 2% aqueous acetic acid gets for 4% chitosan solution;
Described nutrition agent is one or more the mixture in vitriol, nitrate, phosphoric acid salt, the silicate;
Described thickening material is one or more mixtures in xanthan gum, carboxymethyl cellulose, the magnesium aluminum silicate;
Described frostproofer is an ethylene glycol;
Described sanitas is one or more mixtures in Sodium Benzoate, sodium sorbate, the Paraformaldehyde 96;
Described tinting material is sarranine or basic rhodamine.
2. subtilis aqueous suspension type kind clothing according to claim 1 agent, it is characterized in that, described subtilis aqueous suspensions prepares through following steps: earlier with subtilis through liquid submerged fermentation (adopting conventional zymotechnique), centrifugally then remove supernatant, at last with the suitable quantity of water thalline that suspends, make cell concentration in the aqueous suspensions 〉=10
8Individual/mL.
3. subtilis aqueous suspension type kind clothing according to claim 2 agent is characterized in that described subtilis is screened subtilis 69 bacterial strains that get for researchist of the present invention from the natural plant rhizosphere soil.
4. the preparation method of any described subtilis aqueous suspension type kind clothing of claim 1～3 agent is characterized in that, comprises the steps:
(1) with subtilis earlier through liquid submerged fermentation (adopting conventional zymotechnique), centrifugally then remove supernatant, again with the suitable quantity of water thalline that suspends, make cell concentration in the aqueous suspensions 〉=10
8Individual/mL, it is standby to get the subtilis aqueous suspensions;
(2) polyvinyl alcohol is dissolved in 90 ℃ of hot water with 5% weightmeasurement ratio, chitosan is dissolved in 2% aqueous acetic acid with 4% weightmeasurement ratio, according to 8: 2 weight ratio the two is mixed then, make the required film that is mixed into;
(3) in being mixed into film, add nutrition agent, thickening material, frostproofer, sanitas, tinting material, stir, to the subtilis aqueous suspensions of wherein sneaking into step (1), add water more then, stir and promptly get bacillus subtilis suspended seed coating agent of the present invention.
5. the agent of any described subtilis aqueous suspension type kind clothing of claim 1～3 is used to prevent and treat plant soil-borne diseases.
6. the agent of any described subtilis aqueous suspension type kind clothing of claim 1～3 is used to prevent and treat root rot, soybean damping-off.
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| CN200910223355A CN101759491A (en) | 2009-11-19 | 2009-11-19 | Bacillus subtilis suspended seed coating agent, preparation method and application thereof |
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| CN102273485A (en) * | 2011-06-24 | 2011-12-14 | 陈广泉 | Novel corn seed coating agent and manufacturing method thereof |
| CN103004885A (en) * | 2012-11-01 | 2013-04-03 | 无锡亚克生物科技有限公司 | Rape biologic seed coating and preparation method and applications thereof |
| CN103004884A (en) * | 2012-11-01 | 2013-04-03 | 无锡亚克生物科技有限公司 | Biological watermelon seed coating formulation and preparation and applications thereof |
| CN103975955A (en) * | 2014-04-11 | 2014-08-13 | 江苏里下河地区农业科学研究所 | Composite biological seed coating agent used for controlling vegetable soil-borne diseases and preparation and application methods thereof |
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| CN105505836A (en) * | 2016-01-27 | 2016-04-20 | 黑龙江省科学院微生物研究所 | Bacillus subtilis and methods for preparing and using bacillus subtilis microbial agent for controlling potato scab |
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| CN102273485A (en) * | 2011-06-24 | 2011-12-14 | 陈广泉 | Novel corn seed coating agent and manufacturing method thereof |
| CN103004884B (en) * | 2012-11-01 | 2015-08-19 | 无锡亚克生物科技有限公司 | Watermelon biological seed coating agent and Synthesis and applications thereof |
| CN103004885A (en) * | 2012-11-01 | 2013-04-03 | 无锡亚克生物科技有限公司 | Rape biologic seed coating and preparation method and applications thereof |
| CN103004884A (en) * | 2012-11-01 | 2013-04-03 | 无锡亚克生物科技有限公司 | Biological watermelon seed coating formulation and preparation and applications thereof |
| CN103004885B (en) * | 2012-11-01 | 2015-09-02 | 无锡亚克生物科技有限公司 | A kind of brassinosteroid biosynthesis seed coat agent and its production and use |
| CN103975955A (en) * | 2014-04-11 | 2014-08-13 | 江苏里下河地区农业科学研究所 | Composite biological seed coating agent used for controlling vegetable soil-borne diseases and preparation and application methods thereof |
| CN103975955B (en) * | 2014-04-11 | 2016-03-02 | 江苏里下河地区农业科学研究所 | For preventing and treating compound bio seed coat agent and the preparation and application thereof of soil-borne diseases of vegetable |
| CN104430307A (en) * | 2014-08-03 | 2015-03-25 | 石河子大学 | Micro-capsulated suspended microbial seed coating agent and preparation method thereof |
| CN105505836A (en) * | 2016-01-27 | 2016-04-20 | 黑龙江省科学院微生物研究所 | Bacillus subtilis and methods for preparing and using bacillus subtilis microbial agent for controlling potato scab |
| CN105794845A (en) * | 2016-04-15 | 2016-07-27 | 佛山市聚成生化技术研发有限公司 | Microbial pesticide and preparing method thereof |
| CN106635885A (en) * | 2016-11-17 | 2017-05-10 | 黑龙江省农业科学院耕作栽培研究所 | Symbiotic bacterium of entomopathogenic nematode and application of symbiotic bacterium |
| CN106635885B (en) * | 2016-11-17 | 2019-09-20 | 黑龙江省农业科学院耕作栽培研究所 | A kind of entomopathogenic nematode symbiotic bacteria and its application |
| CN108477177A (en) * | 2018-05-16 | 2018-09-04 | 安徽省四达农药化工有限公司 | A kind of difenoconazole suspending seed-dressing agent and preparation method thereof |
| CN112080282A (en) * | 2020-10-12 | 2020-12-15 | 李建勋 | Acidic soil remediation agent and preparation method thereof |
| CN113943184A (en) * | 2021-09-30 | 2022-01-18 | 金禾佳农（北京）生物技术有限公司 | Corn growth-promoting microorganism seed coating agent and coating method thereof |
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Application publication date: 20100630 |