CN103039534A - Microbial preparation for inhibiting root-knot nematode and preparation method and application thereof - Google Patents
Microbial preparation for inhibiting root-knot nematode and preparation method and application thereof Download PDFInfo
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- CN103039534A CN103039534A CN2012105810167A CN201210581016A CN103039534A CN 103039534 A CN103039534 A CN 103039534A CN 2012105810167 A CN2012105810167 A CN 2012105810167A CN 201210581016 A CN201210581016 A CN 201210581016A CN 103039534 A CN103039534 A CN 103039534A
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- 238000002360 preparation method Methods 0.000 title abstract description 12
- 230000002401 inhibitory effect Effects 0.000 title abstract 3
- 241000193386 Lysinibacillus sphaericus Species 0.000 claims abstract description 28
- 241000754833 Pochonia chlamydosporia Species 0.000 claims abstract description 28
- 241001465752 Purpureocillium lilacinum Species 0.000 claims abstract description 28
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims abstract description 23
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- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 claims description 5
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- RRZXIRBKKLTSOM-XPNPUAGNSA-N avermectin B1a Chemical compound C1=C[C@H](C)[C@@H]([C@@H](C)CC)O[C@]11O[C@H](C\C=C(C)\[C@@H](O[C@@H]2O[C@@H](C)[C@H](O[C@@H]3O[C@@H](C)[C@H](O)[C@@H](OC)C3)[C@@H](OC)C2)[C@@H](C)\C=C\C=C/2[C@]3([C@H](C(=O)O4)C=C(C)[C@@H](O)[C@H]3OC\2)O)C[C@H]4C1 RRZXIRBKKLTSOM-XPNPUAGNSA-N 0.000 claims description 4
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Abstract
The invention belongs to the field of biological prevention and control on crop insect damage and provides a microbial preparation for inhibiting root-knot nematode. The preparation is formed by mixing bacterial liquids of streptomyces avermitilis, paecilomyces lilacinus, verticillium chlamydosporium and Bacillus sphaericus and glycerol. The invention also provides a production method and an application effect of the microbial preparation for inhibiting the root-knot nematode. The microbial preparation comprises viable bacteria of which the number is greater than 2,500 millions per milliliter, wherein the viable bacteria comprise the streptomyces avermitilis, the paecilomyces lilacinus, the verticillium chlamydosporium and the Bacillus sphaericus. After the microbial preparation is prepared into soil, eggs of the root-knot nematode are killed by metabolites generated in the continuous growth and reproduction activity process of the viable bacteria, so that a granular structure of the soil is improved, the utilization rate of a fertilizer is improved and an effect of increasing both production and income is achieved.
Description
Technical field
The invention belongs to crop pests biological control field, relate to a kind of microorganism formulation that suppresses root-knot nematode and preparation method thereof and application.
Background technology
Since reported first south China tomato generation root knot nematode disease in 1932, there is the occurrence and harm of root knot nematode disease more than ten provinces and regions, the whole nation.In recent years, China's protection ground vegetables development is rapid, and vegetable crop of the same race is planted in the greenhouse year after year, causes many aged greenhouses root-knot nematode disease to be on the rise, and has caused the significant underproduction of vegetables, has brought hugely economic loss for vegetable growers.Cause 20%-25% economic loss general every year, serious loss 50%-60%, even total crop failure.Therefore, the harm improvement of root-knot nematode become a very important agricultural production task.Nematicide main method is nematocide in the market, as utilizes Celfume, organic phosphorus compound and antibiotic etc. to carry out soil disinfection.Traditional nematocide not only use cost is high, and Environmental compatibility is poor, has destroyed the structure of soil.Along with the formation of green consumption idea with popularize rapidly, in the urgent need to developing the reasonable nematode killing agent of new bio of safety, efficient, low toxicity.
Summary of the invention
The purpose of this invention is to provide a kind of microorganism formulation that suppresses root-knot nematode.
The present invention also aims to provide the preparation method of the microorganism formulation of this inhibition root-knot nematode.
The present invention also aims to provide the effect of the inhibition root-knot nematode of microorganism formulation aspect the agricultural plantings such as vegetables, melon and fruit and tobacco of this inhibition root-knot nematode.
The present invention also aims to use the microorganism formulation of this inhibition root-knot nematode can improve chemical fertilizer utilization ratio, reduce continuous cropping disease and pesticide dosage, improve the safety of crop products.
The invention provides a kind of microorganism formulation that suppresses root-knot nematode, formed by deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus bacterium liquid mixing glycerine, four strain bacterium are antagonism not mutually, and the holding time is long.
Wherein, the component of described each bacterium bacterium liquid by volume percentage be preferably deinsectization streptomycete bacterium liquid 30-40%, Paecilomyces lilacinus liquid 10-20%, Verticillium chlamydosporium 20-30%, Bacillus sphaericus 10-30%, glycerine 20%; The mixed effectively total viable count of each bacterium liquid is greater than 2,500,000,000/milliliter.
The present invention also provides the preparation method of the microorganism formulation of described inhibition root-knot nematode.This preparation method comprises the steps:
(1) respectively the shaking flask cultivation is treated in the original strain activation of deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium, Bacillus sphaericus.
(2) respectively the activated spawn shaking flask of deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus is cultivated, obtain shake-flask seed.
(3) respectively the shake-flask seed of deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus is cultivated into bacterium liquid to be mixed in the amplification of fermentation tank secondary respectively.
(4) deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus bacterium liquid to be mixed mixes in proportion and get final product after the glycerine after will sterilizing and the fermentation.
The activation of deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium is all carried out in culture dish in the described step (1), cultivation temperature is respectively 32 degrees centigrade, 35 degrees centigrade and 32 degrees centigrade, medium is according to the fungi culture medium batching of standard, and incubation time is 1 day; The activation of Bacillus sphaericus is carried out in the test tube slant, and cultivation temperature is 35 degrees centigrade, and medium component is the bacteria culture media of standard, and incubation time is 1 day.
The culture parameters of the shake-flask seed of each bacterial classification is identical with culture parameters in the step (1) in the described step (2), just changes medium into liquid nutrient medium, and the rotating speed that is placed in the shaking flask of 500mL 200 rev/mins is cultivated, and incubation time is 2 days.
The shake-flask seed of deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus amplifies cultivation at the fermentation tank secondary in the described step (3), finger is seeded in the shake-flask seed of the deinsectization streptomycete in the shaking flask, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus respectively in the fermentation tank of 500L and cultivates, and the medium component of four strain bacterium all is the aqueous solution that contains 10% mashed potatoes, 3% bean cake powder.Cultivation temperature is the cultivation temperature of described each bacterial classification of step (1), and the fermentation tank rotating speed is 150 rev/mins, and the throughput coefficient all is 0.5, and incubation time is 27 hours.
The present invention also provides the application method that suppresses the inhibition root-knot nematode of microorganism formulation aspect the agricultural plantings such as vegetables, melon and fruit and tobacco of root-knot nematode, it is characterized in that: evenly splash each usage amount 5L/ mu when ploughing before planting vegetable, melon and fruit and the tobacco.
The present invention also provides the microorganism formulation that suppresses root-knot nematode at the effect that suppresses vegetables, melon and fruit and root knot nematode in tobacco.It is characterized in that: described inhibition root-knot nematode microorganism formulation contains the viable bacteria that comprises deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus greater than 2,500,000,000/ml quantity, continuous growth and breeding by viable bacteria after being manured into soil is movable, secretes an amount of Avermectin, deinsectization element, extracellular enzyme and root system growth-promoting factors.By microbial cell secretion and suppress mechanism by substrate and keep the Avermectin of an amount of concentration and deinsectization element and can kill line eggs in soil and the root system, the incidence of disease of root-knot nematode is reduced more than 85%, avoid using the excessive use of other agricultural chemicals; The granular structure that extracellular enzyme and root system growth-promoting factors by microbial cell secretion can improve soil, thereby the fertilizer nutrient absorption that the preserve moisture and fertility ability that improves soil promotes the prosperity of root system to improve crop simultaneously improves the resistance of crop, can promote increasing both production and income more than 30%.
Description of drawings
Fig. 1: the celery root system of invention product useless contrasts with the celery root system that used the invention product, uses the well developed root system of invention product, and the capillary root is many, the root knot that does not obviously have root-knot nematode to cause.
Embodiment
The component and the volume ratio that suppress each microbial bacterial agent of microorganism formulation of root-knot nematode are: deinsectization streptomycete bacterium liquid 30-40%, Paecilomyces lilacinus liquid 10-20%, Verticillium chlamydosporium 20-30%, Bacillus sphaericus 10-30%, glycerine 20%;
The microorganism formulation of described inhibition root-knot nematode is according to following process steps production:
(1) respectively the original strain of deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium, Bacillus sphaericus is treated the shaking flask cultivation in Microbiological Lab's activation.
Wherein the activation of deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium is all carried out in culture dish, and cultivation temperature is respectively 32 degrees centigrade, and 35 degrees centigrade and 32 degrees centigrade, medium is according to the fungi culture medium batching of standard, and incubation time is 1 day; The activation of Bacillus sphaericus is carried out in the test tube slant, and cultivation temperature is 35 degrees centigrade, and medium component is the bacteria culture media of standard, and incubation time is 1 day.
(2) respectively the activated spawn shaking flask of deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus is cultivated, obtain shake-flask seed.
The culture parameters of the shake-flask seed of each bacterial classification is identical with culture parameters in the step (1), just changes medium into liquid nutrient medium, and the rotating speed that is placed in the shaking flask of 500mL 200 rev/mins is cultivated, and incubation time is 2 days.
(3) respectively the shake-flask seed of deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus is cultivated into bacterium liquid to be mixed in the amplification of fermentation tank secondary respectively.
The shake-flask seed of the deinsectization streptomycete in the shaking flask, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus is seeded in respectively in the fermentation tank of 500L and cultivates, the medium component of four strain bacterium all is the aqueous solution that contains 10% mashed potatoes, 3% bean cake powder.Cultivation temperature is the cultivation temperature of described each bacterial classification of step (1), and the fermentation tank rotating speed is 150 rev/mins, and the throughput coefficient all is 0.5, and incubation time is 27 hours.
(4) deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus bacterium liquid to be mixed mixes in proportion and get final product after the glycerine after will sterilizing and the fermentation.
Embodiment:
With deinsectization streptomycete, the original strain of Paecilomyces lilacinus and Verticillium chlamydosporium utilizes fungi culture medium dull and stereotyped, the original strain of Bacillus sphaericus utilizes the bacteria culture media inclined-plane laboratory activation culture 1 day, with each bacterial classification after the activation respectively in the liquid nutrient medium identical with separately activation culture based component shaking flask cultivated 2 days, then with the cultured bacterial classification of shaking flask respectively in the fermentation tank of 500L with identical all be the medium culture 27 hours of 10% concentration mashed potatoes and 3% bean cake powder, the at last deinsectization streptomycete that the 500L fermentation tank culture is good, the glycerine that Paecilomyces lilacinus and Verticillium chlamydosporium and Bacillus sphaericus and sterilization are good presses 30%, 15%, 25%, 10% and 20% ratio is mixed namely.
Experimental example: carried out respectively the experiment work of the microorganism formulation of this inhibition root-knot nematode in mid-March, 2012 in Da Ping township, Tongcheng County, Hubei Province and Huaihua City An Jiang town, Hunan Province.130 mu of demonstration areas, soil condition, liquid manure and every managerial skills are consistent in the demonstration program.The demonstration field is plants the celery booth for many years, and the root-knot nematode disease is serious.Substantial contribution purchase chemical fertilizer, agricultural chemicals are all cultivated, are are meticulously managed and protected, dropped into to every year meticulously for increasing land fertility and the insect pest of curing the disease of going out in this plot, but along with the growth root-knot nematode of implantation time is more and more severe, occur in recent years dropping in production over a large area more than 40%.After inventing with this this year, surprising variation has occured in the vegetable plot: the plot root-knot nematode of invention product relatively useless obviously suppresses, and celery capillary root is flourishing, and output has improved more than 50% on original basis.(be that the celery root system of this product useless covers with root-knot nematode such as Fig. 1 left side, the capillary root is few; The right does not have nematode substantially for using the celery root system of this invention, well developed root system, and the capillary root is many.）
Claims (4)
1. microorganism formulation that suppresses root-knot nematode, it is characterized in that, mixed in proportion by deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus bacterium liquid and glycerine, four strain bacterium mutual antagonism not under the cooperation of glycerine, and also the holding time can be above 2 years.
2. the microorganism formulation of inhibition root-knot nematode according to claim 1 is characterized in that described deinsectization streptomycete bacterium liquid 30-40%, Paecilomyces lilacinus liquid 10-20%, Verticillium chlamydosporium 20-30%, Bacillus sphaericus 10-30%, glycerine 20%; The mixed effectively total viable count of each bacterium liquid is greater than 2,500,000,000/milliliter.
3. the production method of the microorganism formulation of the described inhibition of claim root-knot nematode, the method comprises the steps:
(1) respectively the shaking flask cultivation is treated in the original strain activation of deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium, Bacillus sphaericus;
(2) respectively the activated spawn shaking flask of deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus is cultivated, obtain shake-flask seed;
(3) respectively the shake-flask seed of deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus is cultivated into bacterium liquid to be mixed in the amplification of fermentation tank secondary respectively;
(4) deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus bacterium liquid to be mixed mixes in proportion and get final product after the glycerine after will sterilizing and the fermentation.
4. the microorganism formulation of the arbitrary described inhibition root-knot nematode of claim 1-3 is in the root-knot nematode inhibition of vegetables, melon and fruit and tobacco, it is characterized in that described inhibition root-knot nematode microorganism formulation contains the viable bacteria that comprises deinsectization streptomycete, Paecilomyces lilacinus, Verticillium chlamydosporium and Bacillus sphaericus greater than 2,500,000,000/ml quantity, continuous growth and breeding by viable bacteria after being manured into soil is movable, secretes an amount of Avermectin, deinsectization element, extracellular enzyme and root system growth-promoting factors; By microbial cell secretion and suppress the Avermectin of an amount of concentration that mechanism keeps and deinsectization element by substrate and can kill line eggs in soil and the root system, the incidence of disease of root-knot nematode is reduced more than 85%, avoid using the excessive use of other agricultural chemicals; The granular structure that extracellular enzyme and root system growth-promoting factors by microbial cell secretion can improve soil, thereby the preserve moisture and fertility ability that improves soil promotes simultaneously the prosperity of root system to improve the fertilizer nutrient absorption of crop and improves the resistance of crop, can promote increasing both production and income more than 30%.
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104371949A (en) * | 2014-08-21 | 2015-02-25 | 北京世纪阿姆斯生物技术股份有限公司 | Root-knot nematode killing composite microbial inoculum and preparation method thereof |
| CN104542727A (en) * | 2015-01-27 | 2015-04-29 | 辽宁玉泉生态农业科技有限公司 | Liquid-state composite biological multi-control insecticide and preparation method thereof |
| CN105689377A (en) * | 2016-03-28 | 2016-06-22 | 曹国平 | Ecological remediation method for soil polluted by nematodes |
| CN105724438A (en) * | 2016-04-12 | 2016-07-06 | 深圳市芭田生态工程股份有限公司 | Functional bactericide for preventing and treating root-knot nematode |
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| CN104371949A (en) * | 2014-08-21 | 2015-02-25 | 北京世纪阿姆斯生物技术股份有限公司 | Root-knot nematode killing composite microbial inoculum and preparation method thereof |
| CN104542727A (en) * | 2015-01-27 | 2015-04-29 | 辽宁玉泉生态农业科技有限公司 | Liquid-state composite biological multi-control insecticide and preparation method thereof |
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