CN103013862A - Method for preparing broad-spectrum growth-promoting biocontrol bacillus B96-II fermentation broth - Google Patents
Method for preparing broad-spectrum growth-promoting biocontrol bacillus B96-II fermentation broth Download PDFInfo
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Abstract
The invention relates to a method for preparing biocontrol strain fermentation broth and particularly relates to a method for preparing broad-spectrum growth-promoting biocontrol bacillus B96-II fermentation broth which can prevent and treat blight-based soil-borne diseases of vegetables and melons and has a growth promoting effect on a variety of vegetables. The method comprises the following steps of: activating a strain; preparing a B96-II strain liquid culture medium; preparing B96-II strain liquid; preparing a B96-II fermentation broth culture medium; and preparing the B96-II fermentation broth. The bacillus B96-II fermentation broth prepared by the method has good broad-spectrum performance, can prevent and treat the diseases of a variety of vegetables and melons, and has a growth promoting effect on a variety of vegetables, so that the bacillus B96-II fermentation broth is widely applicable to the prevention and treatment of the soil-borne diseases of the vegetables and melons.
Description
Technical field
The present invention relates to a kind of making method of biocontrol microorganisms fermented liquid, is specially a kind of melon short preparation method who gives birth to Biocontrol Bacillus B96-II fermented liquid of wide spectrum who various vegetables is had the short fruit of coming into force that reaches take blight as main soil-borne disease of control vegetables that has.
Background technology
Vegetable melon and fruit is the indispensable food of people's daily life, and all people of having have the place in soil that the plantation of vegetable melon and fruit is nearly all arranged, yet the soil-borne diseases such as melon dish blight, damping-off, epidemic disease really accompany with dish all the time.In recent years, along with the continuous progress of industrialized agriculture, for making things convenient for production and marketing transportation, the mode of production of mass-producing big area single cropping grows with each passing day, and for breeding and the diffusion of pathogenic bacteria provides suitable host and environment, causes soil-borne disease to be on the rise.Carrying out soil treatment with chemical agent is the important method of preventing and treating soil-borne disease, the efficient soil fumigant of generally acknowledging in the world such as monobromethane, once be widely used in the control of garden crop soil-borne disease in developed country, but this compound significantly consumes atmospheric ozone layer, has a strong impact on ball ecological environment and human health.Except fumigant, the chemical agent that is used for soil treatment also has quintozene, fenaminosulf etc., but owing to the problems such as poor effect, cost are too high, toxic side effect is too large are difficult to extensive popularization, some in addition disabled.At present, prove through prevention practice that utilizing beneficial microorganism to come the antagonism soil-borne pathogen is long-term and effective means, soil-borne pathogen is fit to field planting and the survival of microorganism because the generation environment is relatively stable, is suitable for and carries out biological control; But the controlling object of existing biological pesticide mostly is simple target, and specific aim is strong, broad spectrum is relatively poor.
Summary of the invention
The present invention is strong for the controlling object specific aim that solves existing microorganism, the relatively poor problem of broad spectrum, provides a kind of wide spectrum the short preparation method who gives birth to Biocontrol Bacillus B96-II fermented liquid.
The present invention adopts following technical scheme to realize: the short preparation method who gives birth to Biocontrol Bacillus B96-II fermented liquid of wide spectrum comprises the steps:
(1) actication of culture: 2-3 is made 3-5 days, temperature in advance for genus bacillus B96-II bacterial classification access remain on 4-10 ℃ PDA slant medium, in 28 ℃ of constant incubators, cultivated 2-3 days, obtain the PDA slant strains;
(2) make B96-II seed liquor substratum: get and clean the potato 200g that peels, cut the 1cm square, the 1L clear water boils warm fire again and boiled 30 minutes, after the filtration filtrate is settled to 1000mL with distilled water, adding potassium primary phosphate 1-10 g, sodium-chlor 1-10g, glucose 10-20g mix rear 121 ℃ of sterilization 20-30 and divide, and obtain B96-II seed liquor substratum;
(3) make the B96-II seed liquor: behind B96-II seed liquor substratum naturally cooling, with the PDA slant strains access that step () makes, 30-34 ℃ of shaking table, 150-180 rev/min, cultivated 24-36 hour, obtain the B96-II seed liquor;
(4) make the B96-II fermentation broth: Semen Maydis powder 1-10g, analysis for soybean powder 1-10g, peptone 1-10g, potassium primary phosphate 1-10g, sodium-chlor 1-15g, ammonium sulfate 1-15g, calcium carbonate 1-10g, sal epsom 0.1-10 g, zinc sulfate 0.1-10g, Manganous chloride tetrahydrate 0.1-10g, yeast extract paste 1-10g, glucose 10-200g, water 1000mL, 121 ℃ of sterilization 20-40 divide, and obtain the B96-II fermentation broth;
(5) make the B96-II fermented liquid: in the B96-II fermentation broth that the B96-II seed liquor that step (three) is made makes by 10-20% volumetric concentration access step (four) (needing the 100-200mLB96-II seed liquor such as the 1000mLB96-II fermentation broth), 30-34 ℃ of shaking table, 150-180 rev/min, cultivated 48 hours, the bacterium liquid that obtains is the B96-II fermented liquid.
Genus bacillus B96-II bacterial strain of the present invention is at non-patent literature---and 06 phase of 15 volumes in 2009 " using and the environmental organism journal ", upper form with paper " Antagonistic Fungi B96-II is to the bacteriostatic action of asparagus blight " was open, public's wish obtains this bacterial strain and can obtain to the application of Shanxi Province's Academy of Agricultural Sciences Shanxi Province agricultural chemicals key lab, the legal holder of this bacterial strain---Shanxi Province's Academy of Agricultural Sciences Shanxi Province agricultural chemicals key lab, this chamber guarantees under the condition of relevant regulations such as being in conformity with law, will provide this bacterial strain to the public in 20 years from the applying date.
Shanxi Province agricultural chemicals key lab screened efficient Antagonistic Fungi since 1996 for soil-borne diseases such as vegetables blight, epidemic diseases.Through up to ten thousand plates, more than 50 inferior greenhouses and field diseases prevention shaker test are proved: genus bacillus B96-II Antagonistic Fungi has obvious restraining effect to 17 kinds of vegetables pathogenic fungi of gourds such as the withered bacterium of asparagus stem, asparagus blight, capsicum wilt, capsicum epidemic disease, tomato wilt, early blight of tomato, cucumber fusarium axysporum, cucumber samping off, watermelon blight, watermelon anthrax, Apple Mould Cores, and the various vegetables such as asparagus, green pepper, capsicum, tomato, celery, cucumber, watermelon, muskmelon are had growth-promoting functions.
The B96-II fermented liquid that makes with the present invention is respectively to the withered bacterium of asparagus stem, the cucumber Fusarium oxysporum, the watermelon Fusarium oxysporum, the green pepper Fusarium oxysporum compares discovery with the control group of only processing with clear water after processing 24h: compare mycelial growth and inhibition of germination with control group after with the B96-II fermented liquid the withered bacterium of asparagus stem being processed and be respectively 89.6% and 99.6%, the inhibiting rate that in vitro tissue is cultivated bacterial plaque is 94.4%, field control effect is 93.40% after 3 months, when the concentration of B96-II fermented liquid is raised to 10% from 1%, specific conductivity (μ S/cm) brings up to 818.3% from 47.5%, total dissolved solidss (TDS) bring up to 797.6%, respiration inhibition rate from 25% to 196.4% from 151.3%; Comparing the germ tube inhibiting rate with control group after with the B96-II fermented liquid cucumber Fusarium oxysporum being processed is 87.5%, and the greenhouse prevention effect is 43.5% and be 53.7% after 3 months after 1 month; Comparing germ tube Germination suppression rate with control group after with the B96-II fermented liquid watermelon Fusarium oxysporum being processed is 77.3%, and the greenhouse prevention effect is 64.6% and be 100% after 3 months after 1 month; Comparing germ tube Germination suppression rate with control group after with the B96-II fermented liquid green pepper Fusarium oxysporum being processed is 78.1%, and the greenhouse prevention effect is 43.1% and be 77.9% after 3 months after 1 month.The experimental result explanation, the B96-II fermented liquid can cause mycelia and the serious breakage of spore, and entocyte is exosmosed; Value with bacterium liquid specific conductivity and total dissolved solidss behind the B96-II fermentation liquor treatment mycelia obviously improves than undressed mycelia, and namely the B96-II fermented liquid can suppress the breathing of pathogenic bacteria mycelia, dissolving mycelia; The greenhouse prevention effect prolongs in time effect and significantly improves; Thereby show that the B96-II fermented liquid has significant prevention effect to the melon soil-borne disease of vegetables.
The B96-II fermented liquid that makes with the present invention is relatively found with untreated control group after cucumber, watermelon, capsicum, asparagus are processed: plant height, fresh weight and dry weight with the cucumber after the B96-II fermentation liquor treatment have increased respectively 18.8%, 55.2% and 46.2; Dry weight with watermelon plant after the B96-II fermentation liquor treatment has increased 20%-30%; Dry weight with the capsicum after the B96-II fermentation liquor treatment has increased about 30%; Dry weight with asparagus after the B96-II fermentation liquor treatment has increased about 20%.The experimental result explanation, with envelope ridge successful between the vegetable fields after the B96-II fermentation liquor treatment, plant is high, fruit is large, urgees to come into force fruit obviously.
The genus bacillus B96-II fermented liquid broad spectrum that the present invention makes is good, can prevent and treat the melon soil-borne disease of various vegetables, and the various vegetables class there is the short fruit of coming into force, greatly reduced production cost, the control of the melon soil-borne disease of vegetables be can be widely used in, industrialized agriculture and the production applications of green non-pollution vegetables such as warmhouse booth are specially adapted to.
Embodiment
Embodiment 1:
The short preparation method who gives birth to Biocontrol Bacillus B96-II fermented liquid of wide spectrum comprises the steps:
(1) actication of culture: 3 days, temperature are made in 3 generation genus bacillus B96-II bacterial classifications accesses in advance remain on 10 ℃ PDA slant medium, in 28 ℃ of constant incubators, cultivated 3 days, obtain the PDA slant strains;
(2) make B96-II seed liquor substratum: get and clean the potato 200g that peels, cut the 1cm square, the 1L clear water boils warm fire again and boiled 30 minutes, after the filtration filtrate is settled to 1000mL with distilled water, adding potassium primary phosphate 5g, sodium-chlor 10g, glucose 15g mix rear 121 ℃ of sterilizations 30 minutes, obtain B96-II seed liquor substratum;
(3) make the B96-II seed liquor: behind B96-II seed liquor substratum naturally cooling, with the PDA slant strains access that step 1 makes, 30 ℃ of shaking tables, 180 rev/mins, cultivated 30 hours, obtain the B96-II seed liquor;
(4) make the B96-II fermentation broth: Semen Maydis powder 10g, analysis for soybean powder 1g, peptone 1g, potassium primary phosphate 6g, sodium-chlor 15g, ammonium sulfate 7g, calcium carbonate 0.1g, sal epsom 10 g, zinc sulfate 6g, Manganous chloride tetrahydrate 10g, yeast extract paste 1g, glucose 105g, water 1000mL, sterilized 40 minutes, and obtained the B96-II fermentation broth for 121 ℃;
(5) make the B96-II fermented liquid: in the B96-II fermentation broth that the B96-II seed liquor that step 3 is made makes by 10% volumetric concentration access step 4,30 ℃ of shaking tables, 165 rev/mins, to cultivate 48 hours, the bacterium liquid that obtains is the B96-II fermented liquid.
Embodiment 2:
The short preparation method who gives birth to Biocontrol Bacillus B96-II fermented liquid of wide spectrum comprises the steps:
(1) actication of culture: 4 days, temperature are made in 2 generation genus bacillus B96-II bacterial classifications accesses in advance remain on 7 ℃ PDA slant medium, in 28 ℃ of constant incubators, cultivated 2 days, obtain the PDA slant strains;
(2) make B96-II seed liquor substratum: get and clean the potato 200g that peels, cut the 1cm square, the 1L clear water boils warm fire again and boiled 30 minutes, after the filtration filtrate is settled to 1000mL with distilled water, adding potassium primary phosphate 10 g, sodium-chlor 1g, glucose 20g mix rear 121 ℃ of sterilizations 25 minutes, obtain B96-II seed liquor substratum;
(3) make the B96-II seed liquor: behind B96-II seed liquor substratum naturally cooling, with the PDA slant strains access that step 1 makes, 34 ℃ of shaking tables, 165 rev/mins, cultivated 36 hours, obtain the B96-II seed liquor;
(4) make the B96-II fermentation broth: Semen Maydis powder 6g, analysis for soybean powder 5g, peptone 10g, potassium primary phosphate 1g, sodium-chlor 8g, ammonium sulfate 1g, calcium carbonate 6g, sal epsom 0.1g, zinc sulfate 0.1g, Manganous chloride tetrahydrate 5g, yeast extract paste 10g, glucose 10g, water 1000mL, sterilized 30 minutes, and obtained the B96-II fermentation broth for 121 ℃;
(5) make the B96-II fermented liquid: in the B96-II fermentation broth that the B96-II seed liquor that step 3 is made makes by 15% volumetric concentration access step 4,32 ℃ of shaking tables, 180 rev/mins, to cultivate 48 hours, the bacterium liquid that obtains is the B96-II fermented liquid.
Embodiment 3:
The short preparation method who gives birth to Biocontrol Bacillus B96-II fermented liquid of wide spectrum comprises the steps:
(1) actication of culture: 5 days, temperature are made in 3 generation genus bacillus B96-II bacterial classifications accesses in advance remain on 4 ℃ PDA slant medium, in 28 ℃ of constant incubators, cultivated 2.5 days, obtain the PDA slant strains;
(2) make B96-II seed liquor substratum: get and clean the potato 200g that peels, cut the 1cm square, the 1L clear water boils warm fire again and boiled 30 minutes, after the filtration filtrate is settled to 1000mL with distilled water, adding potassium primary phosphate 1g, sodium-chlor 6g, glucose 10g mix rear 121 ℃ of sterilizations 20 minutes, obtain B96-II seed liquor substratum;
(3) make the B96-II seed liquor: behind B96-II seed liquor substratum naturally cooling, with the PDA slant strains access that step 1 makes, 32 ℃ of shaking tables, 150 rev/mins, cultivated 24 hours, obtain the B96-II seed liquor;
(4) make the B96-II fermentation broth: Semen Maydis powder 1g, analysis for soybean powder 10g, peptone 5g, potassium primary phosphate 10g, sodium-chlor 1g, ammonium sulfate 15g, calcium carbonate 10g, sal epsom 6 g, zinc sulfate 10g, Manganous chloride tetrahydrate 0.1g, yeast extract paste 6g, glucose 200g, water 1000mL, sterilized 20 minutes, and obtained the B96-II fermentation broth for 121 ℃;
(5) make the B96-II fermented liquid: in the B96-II fermentation broth that the B96-II seed liquor that step 3 is made makes by 20% volumetric concentration access step 4,34 ℃ of shaking tables, 150 rev/mins, to cultivate 48 hours, the bacterium liquid that obtains is the B96-II fermented liquid.
Embodiment 4:
The short preparation method who gives birth to Biocontrol Bacillus B96-II fermented liquid of wide spectrum comprises the steps:
(1) actication of culture: 3.5 days, temperature are made in 3 generation genus bacillus B96-II bacterial classifications accesses in advance remain on 5 ℃ PDA slant medium, in 28 ℃ of constant incubators, cultivated 2 days, obtain the PDA slant strains;
(2) make B96-II seed liquor substratum: get and clean the potato 200g that peels, cut the 1cm square, the 1L clear water boils warm fire again and boiled 30 minutes, after the filtration filtrate is settled to 1000mL with distilled water, adding potassium primary phosphate 9g, sodium-chlor 2g, glucose 12g mix rear 121 ℃ of sterilizations 21 minutes, obtain B96-II seed liquor substratum;
(3) make the B96-II seed liquor: behind B96-II seed liquor substratum naturally cooling, with the PDA slant strains access that step 1 makes, 31 ℃ of shaking tables, 170 rev/mins, cultivated 33 hours, obtain the B96-II seed liquor;
(4) make the B96-II fermentation broth: Semen Maydis powder 2g, analysis for soybean powder 7g, peptone 6g, potassium primary phosphate 4g, sodium-chlor 2g, ammonium sulfate 12g, calcium carbonate 1g, sal epsom 7g, zinc sulfate 4g, Manganous chloride tetrahydrate 2g, yeast extract paste 2g, glucose 30g, water 1000mL, sterilized 22 minutes, and obtained the B96-II fermentation broth for 121 ℃;
(5) make the B96-II fermented liquid: in the B96-II fermentation broth that the B96-II seed liquor that step 3 is made makes by 12% volumetric concentration access step 4,31 ℃ of shaking tables, 122 rev/mins, to cultivate 48 hours, the bacterium liquid that obtains is the B96-II fermented liquid.
Embodiment 5:
The short preparation method who gives birth to Biocontrol Bacillus B96-II fermented liquid of wide spectrum comprises the steps:
(1) actication of culture: 5 days, temperature are made in 2 generation genus bacillus B96-II bacterial classifications accesses in advance remain on 6 ℃ PDA slant medium, in 28 ℃ of constant incubators, cultivated 3 days, obtain the PDA slant strains;
(2) make B96-II seed liquor substratum: get and clean the potato 200g that peels, cut the 1cm square, the 1L clear water boils warm fire again and boiled 30 minutes, after the filtration filtrate is settled to 1000mL with distilled water, adding potassium primary phosphate 8g, sodium-chlor 3g, glucose 16g mix rear 121 ℃ of sterilizations 29 minutes, obtain B96-II seed liquor substratum;
(3) make the B96-II seed liquor: behind B96-II seed liquor substratum naturally cooling, with the PDA slant strains access that step 1 makes, 33 ℃ of shaking tables, 175 rev/mins, cultivated 34 hours, obtain the B96-II seed liquor;
(4) make the B96-II fermentation broth: Semen Maydis powder 3g, analysis for soybean powder 8g, peptone 5g, potassium primary phosphate 7g, sodium-chlor 4g, ammonium sulfate 14g, calcium carbonate 2g, sal epsom 5 g, zinc sulfate 3g, Manganous chloride tetrahydrate 1g, yeast extract paste 3g, glucose 50g, water 1000mL, sterilized 25 minutes, and obtained the B96-II fermentation broth for 121 ℃;
(5) make the B96-II fermented liquid: in the B96-II fermentation broth that the B96-II seed liquor that step 3 is made makes by 17% volumetric concentration access step 4,33 ℃ of shaking tables, 160 rev/mins, to cultivate 48 hours, the bacterium liquid that obtains is the B96-II fermented liquid.
Embodiment 6:
The short preparation method who gives birth to Biocontrol Bacillus B96-II fermented liquid of wide spectrum comprises the steps:
(1) actication of culture: 4.5 days, temperature are made in 3 generation genus bacillus B96-II bacterial classifications accesses in advance remain on 8 ℃ PDA slant medium, in 28 ℃ of constant incubators, cultivated 2 days, obtain the PDA slant strains;
(2) make B96-II seed liquor substratum: get and clean the potato 200g that peels, cut the 1cm square, the 1L clear water boils warm fire again and boiled 30 minutes, after the filtration filtrate is settled to 1000mL with distilled water, adding potassium primary phosphate 7g, sodium-chlor 4g, glucose 13g mix rear 121 ℃ of sterilizations 23 minutes, obtain B96-II seed liquor substratum;
(3) make the B96-II seed liquor: behind B96-II seed liquor substratum naturally cooling, with the PDA slant strains access that step 1 makes, 32 ℃ of shaking tables, 155 rev/mins, cultivated 27 hours, obtain the B96-II seed liquor;
(4) make the B96-II fermentation broth: Semen Maydis powder 5g, analysis for soybean powder 9g, peptone 4g, potassium primary phosphate 8g, sodium-chlor 6g, ammonium sulfate 9g, calcium carbonate 3g, sal epsom 8g, zinc sulfate 1g, Manganous chloride tetrahydrate 6g, yeast extract paste 4g, glucose 150g, water 1000mL, sterilized 28 minutes, and obtained the B96-II fermentation broth for 121 ℃;
(5) make the B96-II fermented liquid: in the B96-II fermentation broth that the B96-II seed liquor that step 3 is made makes by 18% volumetric concentration access step 4,32 ℃ of shaking tables, 170 rev/mins, to cultivate 48 hours, the bacterium liquid that obtains is the B96-II fermented liquid.
Embodiment 7:
The short preparation method who gives birth to Biocontrol Bacillus B96-II fermented liquid of wide spectrum comprises the steps:
(1) actication of culture: 4 days, temperature are made in 3 generation genus bacillus B96-II bacterial classifications accesses in advance remain on 9 ℃ PDA slant medium, in 28 ℃ of constant incubators, cultivated 2.5 days, obtain the PDA slant strains;
(2) make B96-II seed liquor substratum: get and clean the potato 200g that peels, cut the 1cm square, the 1L clear water boils warm fire again and boiled 30 minutes, after the filtration filtrate is settled to 1000mL with distilled water, adding potassium primary phosphate 4g, sodium-chlor 6g, glucose 18g mix rear 121 ℃ of sterilizations 26 minutes, obtain B96-II seed liquor substratum;
(3) make the B96-II seed liquor: behind B96-II seed liquor substratum naturally cooling, with the PDA slant strains access that step 1 makes, 30 ℃ of shaking tables, 160 rev/mins, cultivated 25 hours, obtain the B96-II seed liquor;
(4) make the B96-II fermentation broth: Semen Maydis powder 7g, analysis for soybean powder 3g, peptone 8g, potassium primary phosphate 9g, sodium-chlor 10g, ammonium sulfate 8g, calcium carbonate 8g, sal epsom 4g, zinc sulfate 8g, Manganous chloride tetrahydrate 9g, yeast extract paste 7g, glucose 180g, water 1000mL, sterilized 33 minutes, and obtained the B96-II fermentation broth for 121 ℃;
(5) make the B96-II fermented liquid: in the B96-II fermentation broth that the B96-II seed liquor that step 3 is made makes by 13% volumetric concentration access step 4,30 ℃ of shaking tables, 175 rev/mins, to cultivate 48 hours, the bacterium liquid that obtains is the B96-II fermented liquid.
Embodiment 8:
The short preparation method who gives birth to Biocontrol Bacillus B96-II fermented liquid of wide spectrum comprises the steps:
(1) actication of culture: 3 days, temperature are made in 2 generation genus bacillus B96-II bacterial classifications accesses in advance remain on 7 ℃ PDA slant medium, in 28 ℃ of constant incubators, cultivated 3 days, obtain the PDA slant strains;
(2) make B96-II seed liquor substratum: get and clean the potato 200g that peels, cut the 1cm square, the 1L clear water boils warm fire again and boiled 30 minutes, after the filtration filtrate is settled to 1000mL with distilled water, adding potassium primary phosphate 3g, sodium-chlor 7g, glucose 19g mix rear 121 ℃ of sterilizations 28 minutes, obtain B96-II seed liquor substratum;
(3) make the B96-II seed liquor: behind B96-II seed liquor substratum naturally cooling, with the PDA slant strains access that step 1 makes, 34 ℃ of shaking tables, 173 rev/mins, cultivated 31 hours, obtain the B96-II seed liquor;
(4) make the B96-II fermentation broth: Semen Maydis powder 8g, analysis for soybean powder 4g, peptone 2g, potassium primary phosphate 3g, sodium-chlor 13g, ammonium sulfate 4g, calcium carbonate 9g, sal epsom 2g, zinc sulfate 7g, Manganous chloride tetrahydrate 7g, yeast extract paste 8g, glucose 130g, water 1000mL, sterilized 37 minutes, and obtained the B96-II fermentation broth for 121 ℃;
(5) make the B96-II fermented liquid: in the B96-II fermentation broth that the B96-II seed liquor that step 3 is made makes by 16% volumetric concentration access step 4,34 ℃ of shaking tables, 162 rev/mins, to cultivate 48 hours, the bacterium liquid that obtains is the B96-II fermented liquid.
Claims (1)
1. the short preparation method who gives birth to Biocontrol Bacillus B96-II fermented liquid of wide spectrum is characterized in that: comprise the steps:
(1) actication of culture: 2-3 is made 3-5 days, temperature in advance for genus bacillus B96-II bacterial classification access remain on 4-10 ℃ PDA slant medium, in 28 ℃ of constant incubators, cultivated 2-3 days, obtain the PDA slant strains;
(2) make B96-II seed liquor substratum: get and clean the potato 200g that peels, cut the 1cm square, the 1L clear water boils warm fire again and boiled 30 minutes, after the filtration filtrate is settled to 1000mL with distilled water, adding potassium primary phosphate 1-10 g, sodium-chlor 1-10g, glucose 10-20g mix 121 ℃ of sterilizations of rear bottling 20-30 and divide, and obtain B96-II seed liquor substratum;
(3) make the B96-II seed liquor: behind B96-II seed liquor substratum naturally cooling, with the PDA slant strains access that step () makes, 30-34 ℃ of shaking table, 150-180 rev/min, cultivated 24-36 hour, obtain the B96-II seed liquor;
(4) make the B96-II fermentation broth: Semen Maydis powder 1-10g, analysis for soybean powder 1-10g, peptone 1-10g, potassium primary phosphate 1-10g, sodium-chlor 1-15g, ammonium sulfate 1-15g, calcium carbonate 0.1-10g, sal epsom 0.1-10g, zinc sulfate 0.1-10g, Manganous chloride tetrahydrate 0.1-10g, yeast extract paste 1-10g, glucose 10-200g, water 1000mL, 121 ℃ of sterilization 20-40 divide, and obtain the B96-II fermentation broth;
(5) make the B96-II fermented liquid: in the B96-II fermentation broth that the B96-II seed liquor that step (three) is made makes by 10-20% volumetric concentration access step (four), 30-34 ℃ of shaking table, 150-180 rev/min, to cultivate 48 hours, the bacterium liquid that obtains is the B96-II fermented liquid.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103555609A (en) * | 2013-09-23 | 2014-02-05 | 云南瑞升埃比特生物科技有限公司 | Biocontrol bacillus with effects of resisting diseases and promoting growth and application |
| CN108624527A (en) * | 2018-05-12 | 2018-10-09 | 湖南科技学院 | A kind of ginkgo source growth-promoting preparation of prevention ginger bacterial wilt |
| CN109055270A (en) * | 2018-08-31 | 2018-12-21 | 广东容大生物股份有限公司 | A kind of high-density cultivation method and culture medium for bacillus |
-
2012
- 2012-12-03 CN CN2012105041051A patent/CN103013862A/en active Pending
Non-Patent Citations (4)
| Title |
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| 马利平等: "B96-II对3种枯萎病的防治效果及拮抗物质初步分析", 《华北农学报》 * |
| 马利平等: "拮抗菌B96-II对芦笋枯萎菌的抑菌作用", 《应用与环境生物学报》 * |
| 马利平等: "芦笋茎枯病的生物防治及机理研究", 《中国生态农业学报》 * |
| 马利平等: "芽孢杆菌B96-II对芦笋茎枯病的防治及机制研究", 《华北农学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103555609A (en) * | 2013-09-23 | 2014-02-05 | 云南瑞升埃比特生物科技有限公司 | Biocontrol bacillus with effects of resisting diseases and promoting growth and application |
| CN108624527A (en) * | 2018-05-12 | 2018-10-09 | 湖南科技学院 | A kind of ginkgo source growth-promoting preparation of prevention ginger bacterial wilt |
| CN109055270A (en) * | 2018-08-31 | 2018-12-21 | 广东容大生物股份有限公司 | A kind of high-density cultivation method and culture medium for bacillus |
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