CN104738092A - Composite preparation for preventing and treating cyanosis as well as preparation method and application method of composite preparation - Google Patents
Composite preparation for preventing and treating cyanosis as well as preparation method and application method of composite preparation Download PDFInfo
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Abstract
The embodiment of the invention discloses a composite preparation for preventing and treating cyanosis as well as a preparation method and an application method of the composite preparation, relates to the field of biological preparations and aims at improving the soil quality under the condition that the effect of preventing and treating the cyanosis is good. The composite preparation for preventing and treating the cyanosis comprises the following components in parts by weight: 200-400 parts of lactobacillus plantarum fermented liquid prepared from lactobacillus plantarum, 200-400 parts of bacillus subtilis fermented liquid prepared from bacillus subtilis, 50-100 parts of potassium fulvate and 20-50 parts of chitin, wherein the lactobacillus plantarum is preserved at the China General Microbiological Culture Collection Center on February 20th, 2014, and the preservation number is CGMCC No.8833; and the bacillus subtilis is preserved at the China General Microbiological Culture Collection Center on February 20th, 2014, and the preservation number is CGMCC No.8834. The composite preparation can be used for preventing and treating the cyanosis of plants.
Description
Technical field
The present invention relates to field of biological, particularly relate to a kind of compound formulation preventing and treating verticillium wilt and preparation method thereof, using method.
Background technology
Verticillium wilt is global soil-borne fungus vascular bundle diseases, and being very popular property of morbidity scope is strong, and incidence probability is high, has a strong impact on crop yield and quality.The pathogenic bacteria of verticillium wilt belong to Deuteromycotina on taxonomy, Moniliaceae, and Verticillium dahliae belongs to, and mainly comprises verticillium dahliae (Verticillium dahliae Kleb) and black and white Verticillium dahliae (Verticillium alboatrum).Up-to-date gene order-checking comparative analysis shows, relative to black and white Verticillium dahliae, verticillium dahliae has the gene of more transposons, duplicate factor, participation signal transduction and metabolic regulation, and this just causes verticillium wilt to have variability, easily produces new bion.
Verticillium wilt belongs to soil-borne disease, and germ survives the winter with invalid body with mycelia, chlamydospore in soil, 6-8 of generally can surviving, and contagiosity is extremely strong, administers after infecting without active drug.The host range of verticillium wilt is very wide, and is expanding year by year, and the host existing more than 660 of report plants at present, mainly Malvaceae, Solanaceae, the rose family, composite family, pulse family and Aceraceae plant etc.When after infection process plant root, diffuse to vascular bundle; Due to the Fast-propagation of pathogen, produce a large amount of mycelia and spore, stimulate neighbouring parenchyma cell to produce colloid substance and lignin, obstruction conduit simultaneously, cause Water Transportation difficulty, suppress leaf transpiration effect and photosynthesis, cause plant wither.The obstacle of Water Transportation causes the transport of root nutrition material to be hindered, and then causes blade, bud, fruit early ageing withered.In addition, a kind of wilting toxin containing glycoprotein can also be produced after infection process plant, to CO
2fixing, H
2the photodissociation of O, coenzyme NAD+reduction and membrane passage all have larger toxic action, cause host's vascular bundle functional deterioration, plant metabolism to lack of proper care, cause plant wither withered, crop failure even had no harvest.
This kind of disease of verticillium wilt is the difficult point in agricultural production always, mainly takes the method such as breeding resistant variety, Agro-chemicals control now.The prevention and controls of chemical agent, while controlling disease, without selectively having injured beneficial organism in a large number, having destroyed the microbial bacteria group structure in soil, having caused soil pollution and degeneration.And the method for breeding resistant variety, because verticillium wilt variability is comparatively large and resistant variety seed selection year limit for length resistance is single, control efficiency is not satisfactory.
Summary of the invention
Embodiments of the invention provide a kind of compound formulation preventing and treating verticillium wilt and preparation method thereof, using method, when preventing and treating verticillium wilt and being respond well, can improve soil quality.
For achieving the above object, embodiments of the invention adopt following technical scheme:
A kind of compound formulation preventing and treating verticillium wilt, according to weight proportion, described compound formulation comprises following composition: the Lactobacillus plantarum zymotic fluid be made up of Lactobacillus plantarum 200 parts-400 parts, the fermentation of bacillus subtilis liquid be made up of bacillus subtilis 200 parts-400 parts;
Wherein, described Lactobacillus plantarum (Lactobacillus plantarum) is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on February 20th, 2014, preserving number CGMCC No.8833; Described bacillus subtilis (Bacillus subtilis) is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on February 20th, 2014, preserving number CGMCC No.8834.
Preferably, in the Lactobacillus plantarum zymotic fluid that described Lactobacillus plantarum is made, effective bacterium number is greater than 1 × 10
9cfu/ml, in described fermentation of bacillus subtilis liquid, effective cell concentration is 1-3 × 10
9cfu/ml.
Preferred further, according to described weight proportion, described compound formulation also comprises: potassium fulvate 50 parts-100 parts, chitin 20 parts-50 parts.
A kind of preparation method preventing and treating the compound formulation of verticillium wilt, according to the weight proportion of composition each in above-mentioned compound formulation, Lactobacillus plantarum zymotic fluid and fermentation of bacillus subtilis liquid are mixed by described weight proportion, then potassium fulvate, chitin is added, mix.
Optionally, the preparation method of described Lactobacillus plantarum zymotic fluid comprises:
In the Lactobacillus plantarum pulvis access MRS medium that freeze-drying is preserved, cultivate 24 hours at 37 DEG C; Then 1% inoculum concentration is inoculated in fermentation medium, at 37 DEG C of bottom fermentation 16-18 hour; Wherein, described MRS medium comprises: beef protein powder 10g, flesh of fish extract 10g, yeast extract 10g, glucose 10g, sodium acetate 5g, ammonium citrate 2g, Tween 80 1g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.2g, four water manganese sulphate 0.05g, epsom salt 0.2g, be settled to 1000ml, pH6.2; Described fermentation medium comprises: peptone 10g, dusty yeast 5g, beef extract 10g, glucose 20g, dibasic ammonium citrate 2g, Tween 80 1ml, K
2hPO
42g, MnSO
44H
2o 0.25g, MgSO
47H
2o 0.58g, NaAc 5g, CaCO
315g, is settled to 1000ml, pH6.4;
The preparation method of described fermentation of bacillus subtilis liquid comprises:
Described bacillus subtilis streak inoculation is cultivated 48h in LB solid culture medium 37 DEG C; Oese picking thalline is inoculated in LB liquid medium, 180rpm, 37 DEG C of cultivation 24h; Be inoculated in corn flour dregs of beans medium with 1% inoculum concentration, carry out fermented and cultured; Fermentation condition is: temperature 37 DEG C, tank pressure 0.1Mpa, ventilation 0.8-1vvm, rotating speed 180rpm, and incubation time is 2-3 days, obtains fermentation of bacillus subtilis liquid; Wherein, described LB solid culture medium comprises: tryptone 10g, yeast extract 5g, sodium chloride 10g, agar 20g, deionized water 1000ml, pH7.0; Described corn flour dregs of beans medium comprises: corn flour 2%, bean cake powder 7%, sodium hydrogen phosphate 0.4%, potassium dihydrogen phosphate 0.03%, MgSO
47H
2o 0.1%, defoamer 0.02-0.005%, pH6.8-7.0.
Prevent and treat a using method for the compound formulation of verticillium wilt, comprising:
Described compound formulation is diluted, fill with root or vegetative period complete stool spray.
A kind of compound formulation preventing and treating verticillium wilt that technique scheme provides and preparation method thereof, using method, by Lactobacillus plantarum zymotic fluid, fermentation of bacillus subtilis liquid makes compound formulation, containing active compound bacteria-fermented liquid in this compound formulation, containing the abundant antibacterial material with control of plant disease, comprise lipopeptid class, peptide class, phospholipid, amino acids, nucleic acid, the multiple compounds such as lipopeptide antibiotic Surfactin and polyenoid class material, they can suppress harmful levels of pathogens spore germination, there is killing fungus, bacterium, the effects such as virus and bacterium substance, wherein Surfactin has inhibit activities to various plants disease fungus, its metabolite strong stress resistance, both preventable disease, plant strain growth can be promoted again, also containing multiple exocellular polysaccharide, active enzyme, amino acid, plant hormone, vitamin etc. in zymotic fluid, there is enhancing crop photosynthesis, promote the effect of plant strain growth, Soil Microorganism Bacterial community can also be improved, suppress the effects such as harmful fungoid, bacterium, virus.Therefore the biologic product that the embodiment of the present invention provides when preventing and treating verticillium wilt and being respond well, can promote soil environment.
Preservation illustrates: described Lactobacillus plantarum (Lactobacillus plantarum) is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center CGMCC on February 20th, 2014, and preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City; Preserving number CGMCC No.8833; Described bacillus subtilis (Bacillus subtilis) is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center CGMCC on February 20th, 2014, preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, and preserving number is CGMCC No.8834.
Embodiment
Below the technical scheme in the embodiment of the present invention is clearly and completely described, obviously, described embodiment is only not used in for illustration of the present invention places restrictions on scope of the present invention, and NM specific experiment method in the following example, experimental technique carries out routinely usually.
Embodiments provide a kind of compound formulation preventing and treating verticillium wilt, according to weight proportion, described compound formulation comprises following composition: the Lactobacillus plantarum zymotic fluid be made up of Lactobacillus plantarum 200 parts-400 parts, the fermentation of bacillus subtilis liquid be made up of bacillus subtilis 200 parts-400 parts.
Wherein, described Lactobacillus plantarum (Lactobacillus plantarum) is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center CGMCC on February 20th, 2014, preserving number CGMCC No.8833; Described bacillus subtilis (Bacillus subtilis) is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center CGMCC on February 20th, 2014, preserving number CGMCC No.8834.
Lactobacillus plantarum (Lactobacillus plantarum) described in the embodiment of the present invention and described bacillus subtilis (Bacillus subtilis) are all be separated in the plant rhizosphere soil of robust growth from the rhizosphere soil verticillium wilt of the healthy and strong cotton plants in Shandong Province's Laiwu City verticillium wilt and fusarium wilt plot occurred frequently falls ill serious plot.Described Lactobacillus plantarum called after LP-1.Described bacillus subtilis called after BS-1.
The separating method of above-mentioned two kinds of bacterial strains: plate streak (microbial room of Chinese Academy of Sciences Nanjing soil institute. edaphon organon [M]. Beijing: scientific publication is shut out, 1985), wherein LP-1 is separated used medium is MRS (lactic acid bacteria medium) medium (beef protein powder 10g, flesh of fish extract 10g, yeast extract 10g, glucose 10g, sodium acetate 5g, ammonium citrate 2g, Tween 80 1g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.2g, four water manganese sulphate 0.05g, epsom salt 0.2g, be settled to 1000ml, pH6.2), it is LB medium (tryptone 10g that BS-1 is separated used medium, yeast extract 5g, sodium chloride 10g, agar 20g, deionized water 1000ml, pH7.0).
The authentication method of above-mentioned two kinds of bacterial strains is: morphological feature qualification and the order-checking of 16S rDNA gene fragment order.
BS-1 is accredited as bacillus subtilis (Bacillus subtilis).BS-1 morphological feature: bacillus.Individual cells 0.7-0.8 × 2-3 micron, raw in gemma, oval, cyst does not expand, and without pod membrane, peritrichous, can move, Gram-positive.NA medium culture is after 24 hours, and bacterium colony surfacing is rough, white, and matt, edge is irregular, forms wrinkle mould when liquid culture concentrates growth.
LP-1 is accredited as Lactobacillus plantarum (Lactobacillus plantarum).LP-1 morphological feature: bacterium colony milky, central uplift becomes little blister, rough surface, and edge is irregular.Thalli morphology is: shaft-like, and bar is longer, and two is circle comparatively, paired or catenation.The Physiology and biochemistry assay display of LP-1 bacterial strain, LP-1 can produce lactic acid, cannot fermenting carbohydrate material aerogenesis, without motility, catalase test feminine gender, Gram-positive.
The preparation method of described Lactobacillus plantarum zymotic fluid is: the Lactobacillus plantarum pulvis access MRS medium freeze-drying preserved (beef protein powder 10g, flesh of fish extract 10g, yeast extract 10g, glucose 10g, sodium acetate 5g, ammonium citrate 2g, Tween 80 1g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.2g, four water manganese sulphate 0.05g, epsom salt 0.2g, be settled to 1000ml, pH6.2) 37 DEG C is cultivated 24 hours, is seed culture medium; Then 1% inoculum concentration is inoculated in fermentation medium (peptone 10g, dusty yeast 5g, beef extract 10g, glucose 20g, dibasic ammonium citrate 2g, Tween 80 1ml, K
2hPO
42g, MnSO
44H
2o 0.25g, MgSO
47H
2o 0.58g, NaAc 5g, CaCO
315g, is settled to 1000ml, pH6.4), at 37 DEG C of bottom fermentation 16-18 hour.After fermentation ends, obtain Lactobacillus plantarum zymotic fluid.Preferably, in zymotic fluid, effectively bacterium number is greater than 1 × 10 to need test to ensure
9cfu/ml,
The preparation method of described fermentation of bacillus subtilis liquid is: the Bacillus subtilis strain streak inoculation of preservation is cultivated 48h in LB solid culture medium (tryptone 10g, yeast extract 5g, sodium chloride 10g, agar 20g, deionized water 1000ml, pH7.0) 37 DEG C; Oese picking thalline is inoculated in LB liquid medium, 180rpm, 37 DEG C of cultivation 24h; Corn flour dregs of beans medium (corn flour 2%, bean cake powder 7%, sodium hydrogen phosphate 0.4%, potassium dihydrogen phosphate 0.03%, MgSO is inoculated in 1% inoculum concentration
47H
2o 0.1%, defoamer 0.02-0.005%, pH6.8-7.0), carry out fermented and cultured.Fermentation condition is: temperature 37 DEG C, tank pressure 0.1Mpa, ventilation 0.8-1vvm, rotating speed 180rpm, and incubation time is 2-3 days, obtains fermentation of bacillus subtilis liquid.Preferably, effective cell concentration is 1-3 × 10 to need test to ensure
9cfu/ml.
Preferably, according to described weight proportion, in described compound formulation, also comprise potassium fulvate 50 parts-100 parts, chitin 20 parts-50 parts.
Described chitin has the title of " plant vaccine ", crops can be induced to produce system resistant, increase the resistance of the diseases such as fungi, bacterium, virus, nematode, reduce disease incidence, significantly reduce Pesticide use amount.In addition it also has extremely strong surface attachment performance, can promote that blade is to the absorption of nutriment, strengthens leaf photosynthesis efficiency, promotes plant leaf blade growth.
Described potassium fulvate is that a kind of pure natural mineral matter active potassium element is fertile, the trace element necessary containing plant and multiple somatomedin, full nutrition can be provided to crop, promote the physiological metabolism of various gourd, fruit and vegetable and land for growing field crops crops, promote Root Absorption and plant strain growth, thus strengthen plant resistance, reduce disease and occur and increase crop yield.
Certainly, other materials being conducive to plant healthy growth can also be comprised in described compound formulation, as the material that the those skilled in the art such as growth hormone or gibberellin have a clear understanding of, can add according to the concrete condition of plant, not enumerate at this.
According to producing and fertilising experience, and take into account processing and the many factors in using simultaneously, the embodiment of the present invention additionally provides a kind of preparation method preventing and treating the compound formulation of verticillium wilt, according to the weight proportion of composition each in above-mentioned compound formulation, Lactobacillus plantarum zymotic fluid and fermentation of bacillus subtilis liquid are mixed by described weight proportion, then potassium fulvate, chitin are added, mix.
The embodiment of the present invention additionally provides the using method of above-mentioned compound formulation, comprising: diluted by described compound formulation, fill with root or vegetative period complete stool spray.
The biologic product that the embodiment of the present invention provides is universal, pollution-free, can be widely used in the soil-borne disease controls such as the verticillium wilt of the crops such as cotton, eggplant, capsicum, tomato, fusarium wilt.
(composition is that in zymotic fluid, effective bacterium number is greater than 1 × 10 to the following provide above-mentioned a kind of compound formulation
9the Lactobacillus plantarum zymotic fluid of cfu/ml, effectively cell concentration are 1-3 × 10
9the fermentation of bacillus subtilis liquid of cfu/ml, chitin, potassium fulvate) 4 embody rule test on verticillium wilt plant, the effect of described compound formulation is described.
Test 1: described compound formulation is to the prophylactic tria (Shouguang, Shandong) of eggplant verticillium wilt
Examination Eggplant Varieties is supplied to be the green eggplant in Xi'an, conventional field planting management.When seedling grow to four leaves wholeheartedly time, the seedling choosing 150 strain growing ways consistent is divided into 3 groups, uses the product of the present invention of different extension rate to carry out filling root, every strain 50mL, processes 2 times, 3 days, interval, to water the clear water of same volume for control group.Root method inoculation verticillium wilt pathogen was hindered to plant employing in after last process the 3rd day, the concentration of spore suspension is 1 × 107/L, every strain inoculation 50mL, within the 4th day after connecing bacterium, institute an inquiry incidence, every investigation in 5 days 1 time, continuous observation 5 times, when control group morbidity is more serious, calculates the incidence of disease, disease index and control efficiency.Result is as shown in table 1, and after the compound formulation using the embodiment of the present invention to provide, the eggplant verticillium wilt incidence of disease and disease index significantly reduce, and preventive effect reaches 89.67%.
Table 1 is the prophylactic tria result of described compound formulation to eggplant verticillium wilt
Table 1
Test 2: described compound formulation occurs and the impact (Heze City, Shandong Province) of output cotton verticillium wilt
For the old cotton field that examination field is plant cotton continuously for many years, verticillium wilt morbidity is serious.Supply examination soil to be moisture soil, soil organic matter 11.28g/kg, full nitrogen 1.45g/kg, alkali-hydrolyzable nitrogen 104.5mg/kg, rapid available phosphorus 12.1mg/kg, available potassium 107.5mg/kg, cotton variety is that Shandong cotton grinds No. 28, and field management is local modus operandi.Test establishes 3 processed group, processed group 1: the present invention's 400 times of dilution seed soaking, and sprays in bud stage complete stool; Processed group 2: the present invention's 800 times of dilution seed soaking, and spray in bud stage complete stool; Control group 3: use clear water process the same period.Result is as shown in table 2, and the present invention has good control efficiency for the cotton verticillium wilt in verticillium wilt plot occurred frequently, reduces the incidence of disease and the disease index of cotton verticillium wilt, improves lint yield.
Table 2 occurs and the impact of output cotton verticillium wilt for the present invention
Table 2
Test 3: described compound formulation is on tomato verticillium wilt, bacterial wilt morbidity and the impact (Jinan, Shandong Province) of output
Test plot soil is yellowish soil, content of organic matter 10.6g/kg, alkali-hydrolyzable nitrogen 76.4mg/kg, rapid available phosphorus 12.5mg/kg, and available potassium 93.6mg/kg, PH are 6.5, and tomato variety is that gingko is rich by force, by line-spacing 35cm, spacing in the rows 30cm, and ridge height 15cm, Routine Management.Apply product formulation of the present invention, with the many bacterium of commercially available bactericide clearly compared with, on the impact of tomato yield and quality.Test establishes 3 process, processed group 1: the compound formulation that the embodiment of the present invention provides 800 times of dilutions spray in tomato seedling phase, bud stage, the phase complete stool of bearing fruit; Processed group 2: the clear 800 times of dilution usings method of many bacterium are with processed group 1; Control group: use clear water process the same period.Result is as shown in table 3, can reduce the incidence of disease of tomato verticillium wilt, bacterial wilt after using this product, increases tomato yield, shortens Time To Market.
Table 3 is the impact of this product on tomato verticillium wilt, the bacterial wilt incidence of disease and output
Table 3
Test 4: described compound formulation is on the impact (Zhangqiu in Shandong province) of capsicum economical character and output
Test plot soil is yellowish soil, and content of organic matter 10.6g/kg, alkali-hydrolyzable nitrogen 66.8mg/kg, rapid available phosphorus 15.3mg/kg, available potassium 78.1mg/kg, PH are 6.6.Supplying to study thing is capsicum, and kind is the rich early green pepper No. 8 of gold, and fertilizing management pattern is local usual way.3 process are established in test altogether, are respectively clear water contrast (control group), the present invention 800 times dilution liquid irrigating root (processed group 1), hymexazol 1000 times dilution liquid irrigating root (processed group 2).Result is as shown in table 4, and after spraying the present invention, capsicum verticillium wilt, the gray mold incidence of disease obviously decline, and yield of hot pepper improves, and in fruit, vitamin C and sugared content obviously increase, and improve quality and the mouthfeel of capsicum.
Table 4 is foliage fertilizer on the impact of capsicum economical character and output
Table 4
The above; be only the specific embodiment of the present invention, but protection scope of the present invention is not limited thereto, is anyly familiar with those skilled in the art in the technical scope that the present invention discloses; the change that can expect easily or replacement, all should be encompassed within protection scope of the present invention.Therefore, protection scope of the present invention should described be as the criterion with the protection domain of claim.
Claims (6)
1. prevent and treat the compound formulation of verticillium wilt for one kind, it is characterized in that, according to weight proportion, described compound formulation comprises following composition: the Lactobacillus plantarum zymotic fluid be made up of Lactobacillus plantarum 200 parts-400 parts, the fermentation of bacillus subtilis liquid be made up of bacillus subtilis 200 parts-400 parts;
Wherein, described Lactobacillus plantarum (Lactobacillus plantarum) is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preserving number CGMCCNo.8833 on February 20th, 2014; Described bacillus subtilis (Bacillus subtilis) is preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center on February 20th, 2014, preserving number CGMCC No.8834.
2. compound formulation according to claim 1, is characterized in that, in the Lactobacillus plantarum zymotic fluid that described Lactobacillus plantarum is made, effective bacterium number is greater than 1 × 10
9cfu/ml, in described fermentation of bacillus subtilis liquid, effective cell concentration is 1-3 × 10
9cfu/ml.
3. compound formulation according to claim 1 and 2, is characterized in that, according to described weight proportion, described compound formulation also comprises: potassium fulvate 50 parts-100 parts, chitin 20 parts-50 parts.
4. prevent and treat the preparation method of the compound formulation of verticillium wilt for one kind, it is characterized in that, the weight proportion of each composition in compound formulation according to claim 3, Lactobacillus plantarum zymotic fluid and fermentation of bacillus subtilis liquid are mixed by described weight proportion, then potassium fulvate, chitin are added, mix.
5. preparation method according to claim 4, is characterized in that, the preparation method of described Lactobacillus plantarum zymotic fluid comprises:
In the pulvis access MRS medium of the Lactobacillus plantarum that freeze-drying is preserved, cultivate 24 hours at 37 DEG C; Then 1% inoculum concentration is inoculated in fermentation medium, at 37 DEG C of bottom fermentation 16-18 hour; Wherein, described MRS medium comprises: beef protein powder 10g, flesh of fish extract 10g, yeast extract 10g, glucose 10g, sodium acetate 5g, ammonium citrate 2g, Tween 80 1g, dipotassium hydrogen phosphate 2g, magnesium sulfate 0.2g, four water manganese sulphate 0.05g, epsom salt 0.2g, be settled to 1000ml, pH6.2; Described fermentation medium comprises: peptone 10g, dusty yeast 5g, beef extract 10g, glucose 20g, dibasic ammonium citrate 2g, Tween 80 1ml, K
2hPO
42g, MnSO
44H
2o 0.25g, MgSO
47H
2o 0.58g, NaAc 5g, CaCO
315g, is settled to 1000ml, pH6.4;
The preparation method of described fermentation of bacillus subtilis liquid comprises:
Described bacillus subtilis streak inoculation is cultivated 48h in LB solid culture medium 37 DEG C; Oese picking thalline is inoculated in LB liquid medium, 180rpm, 37 DEG C of cultivation 24h; Be inoculated in corn flour dregs of beans medium with 1% inoculum concentration, carry out fermented and cultured; Fermentation condition is: temperature 37 DEG C, tank pressure 0.1Mpa, ventilation 0.8-1vvm, rotating speed 180rpm, and incubation time is 2-3 days, obtains fermentation of bacillus subtilis liquid; Wherein, described LB solid culture medium comprises: tryptone 10g, yeast extract 5g, sodium chloride 10g, agar 20g, deionized water 1000ml, pH7.0; Described corn flour dregs of beans medium comprises: corn flour 2%, bean cake powder 7%, sodium hydrogen phosphate 0.4%, potassium dihydrogen phosphate 0.03%, MgSO
47H
2o 0.1%, defoamer 0.02-0.005%, pH6.8-7.0.
6. prevent and treat a using method for the compound formulation of verticillium wilt, it is characterized in that, described compound formulation is diluted, fill with root or vegetative period complete stool spray.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109320368A (en) * | 2018-11-08 | 2019-02-12 | 北京泰克美高新技术有限公司 | The biological agent and preparation method thereof for preventing and treating liriomyza bryoniae |
| CN112400906A (en) * | 2021-01-04 | 2021-02-26 | 河南比赛尔农业科技有限公司 | Rape plant waste bacterium powder preparation for preventing and treating plant root diseases and preparation method and application thereof |
| CN115417720A (en) * | 2022-09-13 | 2022-12-02 | 山东农大肥业科技股份有限公司 | Liquid compound microbial fertilizer with long shelf life and no barrel expansion and preparation method thereof |
-
2014
- 2014-05-04 CN CN201410180962.XA patent/CN104738092A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109320368A (en) * | 2018-11-08 | 2019-02-12 | 北京泰克美高新技术有限公司 | The biological agent and preparation method thereof for preventing and treating liriomyza bryoniae |
| CN112400906A (en) * | 2021-01-04 | 2021-02-26 | 河南比赛尔农业科技有限公司 | Rape plant waste bacterium powder preparation for preventing and treating plant root diseases and preparation method and application thereof |
| CN115417720A (en) * | 2022-09-13 | 2022-12-02 | 山东农大肥业科技股份有限公司 | Liquid compound microbial fertilizer with long shelf life and no barrel expansion and preparation method thereof |
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