CN101843260A - Method for preparing compound bactericide for preventing and controlling soybean root rot - Google Patents
Method for preparing compound bactericide for preventing and controlling soybean root rot Download PDFInfo
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- CN101843260A CN101843260A CN200910010876A CN200910010876A CN101843260A CN 101843260 A CN101843260 A CN 101843260A CN 200910010876 A CN200910010876 A CN 200910010876A CN 200910010876 A CN200910010876 A CN 200910010876A CN 101843260 A CN101843260 A CN 101843260A
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Abstract
The invention belongs to the technological field of agricultural production, and particularly relates to a method for preparing compound bactericide for preventing and controlling soybean root rot. The method comprises the following steps of: collecting field healthy soybean root soil, and specifically screening pseudomonas and bacillus for later use by using a King'B culture medium and a method for coating a beef extract peptone culture medium after 10 minutes of water bath at the temperature of 80 DEG C respectively; collecting roots of a field soybean root rot plant, and separating pathogenic fungi by using a martin culture medium; and re-screening bacterial strains for efficiently inhibiting the activity of the screened pathogenic fungi from the pseudomonas and the bacillus for the lateral use, fermenting the bacterial strains respectively, and then compounding the fermented bacterial strains according to a proportion to obtain the compound bactericide for preventing and controlling the soybean root rot. The screening method for preventing mixed bacteria by using soybean root growth can effectively improve the bioactivity and expand the control range; the compound bactericide is from the healthy soybean roots and most from soybean probiotics; and the colonization can be effectively performed on the soybean roots, so the utilization rate of the bactericide is enhanced.
Description
Technical field
The invention belongs to the agricultural production sciemtifec and technical sphere, a kind of specifically preparation prevents and treats the method for the composite bacteria agent capable of root rot.
Background technology
Root rot is a kind of worldwide disease serious, that the pathogen kind is various and control is difficult that distributes extensively, endangers.Root rot mainly is distributed in Spring Sowing Soybean in Northern and southern the Yellow River and Huai He River summer soybean production area in China, wherein especially takes place serious with area, northeast, Heilungkiang.Root rot main harm soybean root system, influence crop to moisture and nutrient absorbing.The generation of this disease can make that the root of soybean plant strain is heavy, root nodule heavy, nitrogenase activity and lateral root number average are subjected to influence in various degree.According to investigations should the general underproduction 10-30% of disease, can reach 60%-90% when serious, even total crop failure, and the soybean oil content is obviously descended.
The pathogen kind of root rot is various, be mainly: sickle-like bacteria (Fusarium spp.), Rhizoctonia solani Kuhn (Rhizoctonia Solani), pythium spp (Pythium spp.) and phytophthora (Phytophthora spp.) etc., wherein sickle-like bacteria is a dominant bacteria, disease is the most general, improve gradually with the seed sprouting infection rate, during the harm soybean, root begins variable color from the tip of a root, be water soaking mode, the main root Lower Half shows the brown striped, enlarge gradually later on, epidermis and cortex blackening are rotted, and the main root Lower Half rots when serious.Blade is flavescence gradually from bottom to top, and plant is downgraded, and dross is few, and plant death endangers very serious when serious.During local fast ponding, pythium spp and phytophthora disease are heavier.
At present, control to this disease, except some agro-farming measures (as: with the gramineous crop efficent rotation, in good time late sowing, upper seeding, select for use and cultivate disease-resistant kind etc.) outside, mainly be the control of chemical agent, as carbendazim, tmtd, many gram good fortune, happy in good time etc., though dressing seed can be postponed the phase of infecting, but the general soybean chemical control root rot time is short, drug effect is poor, because root rot generally can continue about two months, and the general medicament term of validity had only about one month, later stage, pathogen also can infect soybean, and use chemical agent for a long time, pathogen can produce the resistance variety, and difficulty of prevention and cure is strengthened.
Utilizing biologic product to prevent and treat root rot is present research focus, because biologic product is with strong points, pollution-free, the term of validity is long.Utilization be effective outlet of biocontrol fungicide, but there is the field planting difficulty in biocontrol fungicide always to the antagonism screening biocontrol bacteria of disease fungus, and is active low, to characteristics such as the root pathogens specific aim are not strong, limited development.The reasonable root rot biocontrol fungicide of usefulness mainly is that biologic product dust loam is secreted (a kind of actinomycetes granule) at present, control sickle-like bacteria and purple Penicillium notatum; Also have the Paecilomyces lilacinus microbial inoculum: its secretion has inhibitory action to sickle-like bacteria, but effectively the biocontrol bacteria microbial inoculum is also seldom reported.
Summary of the invention
The object of the invention is to provide a kind of preparation to prevent and treat the method for the composite bacteria agent capable of root rot.
For achieving the above object, the technical solution used in the present invention is: the earth bacteria suspension fetches earth, it is the field soybean rhizosphere soil, utilize the method specificity screening pseudomonad (Pseudomonas spp.) and the bacillus (Bacillus spp.) that are coated with beef-protein medium behind King ' B medium and the 80 ℃ of water-bath 10min respectively, stand-by; Other gathers root rot diseased plant root, utilizes Martin's medium bacterial isolate fungi; Adopt the method for the dull and stereotyped Oxford of PDA cup face-off, bacterial strain with disease fungus activity that efficient inhibition screens, the composite composite bacteria agent capable that can obtain preventing and treating root rot after the fermentation process respectively will be sifted out in the pseudomonad of above-mentioned specificity screening and the bacillus again.
Described King ' B nutrient media components is: show peptone 20g, potassium dihydrogen phosphate 1.2g, epsom salt 1.5g, glycerine 10ml, agar 15g, distilled water 1000ml, pH=7.4-7.6, mix the back sterilization, the sterilization back adds ampicillin, chloramphenicol and the cycloheximide of 0.22um membrane filtration degerming.Described ampicillin is 40ug/ml with the sterile water concentration of ordinary dissolution; Chloramphenicol is 13ug/ml with 95% dissolve with ethanol concentration; Cycloheximide is 100ug/ml with acetone solution concentration.
Described beef-protein medium component is: beef extract 5g, peptone 10g, sodium chloride 5g, agar 15g, distilled water 1000ml, pH=7.0-7.2.Soil supension temperature in 80 ℃ of water-baths was bathed 10 minutes, and dilution is coated with flat board again.Soil supension temperature in 80 ℃ of water-baths was bathed 10 minutes, and dilution is coated with flat board again.
The prescription of described Martin's medium is: glucose 10g, peptone 5g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, 0.1% rose-bengal solution 3.3ml, agar 20g, distilled water 1000ml, pH nature.The sterilization back adds 80% lactic acid 2ml.
Solid PDA culture medium prescription used in the method for the dull and stereotyped face-off of described employing Oxford cup is as follows: potato peeling 200g, sucrose 20g, distilled water 1000ml, pH nature.Potato is cut into small pieces, and adds distilled water and boils 20min, filters, and filtrate adds sucrose, and constant volume adds 20g agar, 115 ℃ of autoclaving 30min to 1000ml.
The method of the dull and stereotyped Oxford of PDA cup face-off, disease fungus mycelia piece at the inoculation diameter 8mm of murphy juice solid plate central authorities, place the Oxford cup after the calcination on the flame in the position of distance mycelia piece 3cm, perhaps the mycelia piece was cultivated after 1 day, place the Oxford cup again, add the cultured products of each stand-by pseudomonad or bacillus in the cup of Oxford, cultivate after 2-3 days, detect the bacteriostasis rate of each bacterium.
With pseudomonad through sifting out again and bacillus mixed culture fermentation respectively, the hybrid bacterial strain of pseudomonas is fermented and cultured in above-mentioned King ' the B medium of added with antibiotic not; The hybrid bacterial strain of bacillus carries out fermentation medium in beef-protein medium; The thalline quantity for the treatment of two genus all reach 1,000,000,000/more than the ml, use mass ratio 2: 8-1 respectively: absorption is air-dry and pulverize down at 40-50 ℃ for 9 wheat bran and turfy soil, and then with mass ratio 3: 7-5: 5 ratio is mixed, and makes solid fungicide; Or respectively with the zymocyte liquid of two genus all the simmer down to original volume 1/2, then by volume 3: 7-5: 5 ratio is mixed, and liquid bacterial agent is made in aseptic packaging.
The condition of pseudomonad fermentation tank is; The throughput of filtrated air is 1: 0.8-1, and mixing speed is 180-200 commentaries on classics/min, and cultivation temperature is 28-30 ℃, and incubation time is 20-30h; And the fermentation condition of bacillus is: the throughput of filtrated air is 1: 0.8-1, and mixing speed is 180-200 commentaries on classics/min, and cultivation temperature is 28-30 ℃, and incubation time is 40-60h, helps forming gemma.
The advantage that the present invention had:
1. the present invention is the degradation bacteria of preventing and treating root rot of substrate from the screening of original position rhizosphere with the soybean rhizosphere, and its field planting ability at the soybean rhizosphere is stronger, can effectively protect the soybean rhizosphere to avoid infecting of disease fungus.
2. the present invention utilizes resistant microorganism to control the disease fungus of root rot according to the principle of microorganism antagonism, and with strong points, the disease-resistant cycle is long, and environmental protection.
3. it utilizes soybean rhizosphere biological and ecological methods to prevent plant disease, pests, and erosion mixed cell method for screening of the present invention, effectively improves biologically active and prevention and treatment range, and its source mostly is the soybean probio, and can grows effectively surely at the soybean rhizosphere for healthy soybean rhizosphere, has strengthened the availability of microbial inoculum.
5. pseudomonad can secrete 2,4-diacetyl phloroglucin, and the azophenlyene carboxylic acid, pyrrolnitrin, antibiotics or other active substances such as the green dense rhzomorph of gamboge are having significant contribution aspect the control silborne fungal diseases.Bacillus can secrete economic benefits and social benefits rhzomorph isoreactivity material can effectively suppress disease fungies such as sickle-like bacteria, so the present invention screens the bacterial classification of this two bacterioid as composite bacteria agent capable, can prevent and treat root rot like this, can solve the field planting problem again.In the hope of reaching the effect of good biological and ecological methods to prevent plant disease, pests, and erosion root rot disease.
Embodiment
Embodiment 1
Gather the healthy soybean rhizosphere soil (Fuyuan County soybean field, Kiamusze City, Heilongjiang Province) in field, 10g soil adds the 90ml distilled water diluting and becomes bacteria suspension, utilize King ' B solid culture medium, ampicillin (the 40ug/ml that adds the degerming of 0.22um membrane filtration behind the medium sterilization, the sterile water dissolving), chloramphenicol (13ug/ml 95% dissolve with ethanol), cycloheximide (100ug/ml acetone solution), specificity screening pseudomonad (Pseudomonas spp.);
Bacteria suspension temperature in 80 ℃ of water-baths after the above-mentioned soil sample dilution is bathed 10min, utilizes beef extract-peptone solid culture medium screening bacillus (Bacillus spp.) then.
Gather field soybean root rot diseased plant root (Fuyuan County soybean field, Kiamusze City, Heilongjiang Province), the broken tissue in laboratory, (the sterilization back adds 2ml lactic acid with Martin's medium, suppressing bacterium and actinomycetic growth) the bacterial isolate fungi is mainly Fusarium oxysporum (Fusarium oxysporum), Fusarium graminearum (Fusarium graminearum) and Rhizoctonia solani Kuhn (RhizoctoniaSolani), and is stand-by.
Adopt the method for the dull and stereotyped Oxford of murphy juice (PDA) cup face-off, that is: with the disease fungus of above-mentioned screening respectively in inoculated and cultured on the PDA solid culture medium after 3 days, get the mycelia piece of diameter 8mm at the mycelia edge with the card punch of sterilizing, standby; Inoculate the mycelia piece respectively in new PDA solid plate central authorities, cultivate after 1 day, place the Oxford cup in the position of distance mycelia piece 3cm (on the flame about calcination 5s, place again), the cultured products that adds each pseudomonad or bacillus in the cup of Oxford was respectively cultivated after 2 days, detected the bacteriostasis rate of each bacterium.
Intend: the bacteriostasis rate of positive control is 100%, and the bacteriostasis rate of negative control is 0,
Filter out 36 pseudomonas and 21 bacillus with disease fungus activity that efficient inhibition screens, the mixed culture fermentation respectively of pseudomonad and bacillus, the condition of pseudomonad fermentation tank is: the throughput of filtrated air is 1: 0.8, mixing speed is 180 commentaries on classics/min, cultivation temperature is 28 ℃, and incubation time is 30h; And the fermentation condition of bacillus is: the throughput of filtrated air is 1: 0.8, and mixing speed is 180 commentaries on classics/min, and cultivation temperature is 28 ℃, and incubation time is 60h, and microscopically is observed most of gemma that forms.After the fermentation ends, the thalline quantity of two genus all reach 1,000,000,000/more than the ml, down absorption is air-dry and pulverize at 45 ℃ to use wheat bran and turfy soil (mass ratio 2: 8) respectively, so latter two belongs to 3: 7 ratio of mass ratio and mixing, and makes solid fungicide; Or respectively with 1/2 of the zymocyte liquid simmer down to original volume of two genus, 3: 7 by volume ratio is mixed then, and liquid bacterial agent is made in aseptic packaging.
Embodiment 2
Gather the healthy soybean rhizosphere soil (Fuyuan County soybean field, Kiamusze City, Heilongjiang Province) in field, 10g soil adds the 90ml distilled water diluting and becomes bacteria suspension, utilize King ' B solid culture medium, ampicillin (the 40ug/ml that adds the degerming of 0.22um membrane filtration behind the medium sterilization, the sterile water dissolving), chloramphenicol (13ug/ml 95% dissolve with ethanol), cycloheximide (100ug/ml acetone solution), specificity screening pseudomonad (Pseudomonas spp);
Bacteria suspension temperature in 80 ℃ of water-baths after the above-mentioned soil sample dilution is bathed 10min, utilizes beef extract-peptone solid culture medium screening bacillus (Bacillus spp.) then.
Gather field soybean root rot diseased plant root (Fuyuan County soybean field, Kiamusze City, Heilongjiang Province), the broken tissue in laboratory, (the sterilization back adds 2ml lactic acid with Martin's medium, suppressing bacterium and actinomycetic growth) the bacterial isolate fungi is mainly Fusarium oxysporum (Fusarium oxysporum), Fusarium graminearum (Fusarium graminearum) and Rhizoctonia solani Kuhn (RhizoctoniaSolani), and is stand-by.
Adopt the method for the dull and stereotyped Oxford of murphy juice (PDA) cup face-off, that is: with the disease fungus of above-mentioned screening respectively in inoculated and cultured on the PDA solid culture medium after 3 days, get the mycelia piece of diameter 8mm at the mycelia edge with the card punch of sterilizing, standby; Inoculate the mycelia piece respectively in new PDA solid plate central authorities, place Oxford cup (, placing again) in the position of distance mycelia piece 3cm, add the cultured products of each pseudomonad or bacillus in the cup of Oxford respectively on the flame about calcination 5s, cultivate after 3 days, detect the bacteriostasis rate of each bacterium.
Intend: the bacteriostasis rate of positive control is 100%, and the bacteriostasis rate of negative control is 0,
Filter out 36 pseudomonas and 21 bacillus with disease fungus activity that efficient inhibition screens, the mixed culture fermentation respectively of pseudomonad and bacillus, the condition of pseudomonad fermentation tank is: the throughput of filtrated air is 1: 0.8, mixing speed is 180 commentaries on classics/min, cultivation temperature is 28 ℃, and incubation time is 30h; And the fermentation condition of bacillus is: the throughput of filtrated air is 1: 0.8, and mixing speed is 180 commentaries on classics/min, and cultivation temperature is 28 ℃, and incubation time is 60h, the most of gemma that forms of the little observation of microscope.After the fermentation ends, the thalline quantity of two genus all reach 1,000,000,000/more than the ml, down absorption is air-dry and pulverize at 50 ℃ to use wheat bran and turfy soil (mass ratio 1: 9) respectively, mixes with 5: 5 ratio of mass ratio then, makes solid fungicide; Or respectively with 1/2 of the zymocyte liquid simmer down to original volume of two genus, 5: 5 by volume ratio is mixed then, and liquid bacterial agent is made in aseptic packaging.
Embodiment 3
Gather the healthy soybean rhizosphere soil (Fuyuan County soybean field, Kiamusze City, Heilongjiang Province) in field, 10g soil adds the 90ml distilled water diluting and becomes bacteria suspension, utilize King ' B solid culture medium, ampicillin (the 40ug/ml that adds the degerming of 0.22um membrane filtration behind the medium sterilization, the sterile water dissolving), chloramphenicol (13ug/ml 95% dissolve with ethanol), cycloheximide (100ug/ml acetone solution), specificity screening pseudomonad (Pseudomonas spp.);
Bacteria suspension temperature in 80 ℃ of water-baths after the above-mentioned soil sample dilution is bathed 10min, utilizes beef extract-peptone solid culture medium screening bacillus (Bacillus spp.) then.
Gather field soybean root rot diseased plant root (Fuyuan County soybean field, Kiamusze City, Heilongjiang Province), the broken tissue in laboratory, (the sterilization back adds 2ml lactic acid with Martin's medium, suppressing bacterium and actinomycetic growth) the bacterial isolate fungi is mainly Fusarium oxysporum (Fusarium oxysporum), Fusarium graminearum (Fusarium graminearum) and Rhizoctonia solani Kuhn (RhizoctoniaSolani), and is stand-by.
Adopt the method for the dull and stereotyped Oxford of murphy juice (PDA) cup face-off, that is: with the disease fungus of above-mentioned screening respectively in inoculated and cultured on the PDA solid culture medium after 3 days, get the mycelia piece of diameter 8mm at the mycelia edge with the card punch of sterilizing, standby; Inoculate the mycelia piece respectively in new PDA solid plate central authorities, cultivate after 1 day, place the Oxford cup in the position of distance mycelia piece 3cm (on the flame about calcination 5s, place again), the cultured products that adds each pseudomonad or bacillus in the cup of Oxford was respectively cultivated after 2 days, detected the bacteriostasis rate of each bacterium.
Intend: the bacteriostasis rate of positive control is 100%, and the bacteriostasis rate of negative control is 0,
Filter out 36 pseudomonas and 21 bacillus with disease fungus activity that efficient inhibition screens, the mixed culture fermentation respectively of pseudomonad and bacillus, the condition of pseudomonad fermentation tank is: the throughput of filtrated air is 1: 1, mixing speed is 200 commentaries on classics/min, cultivation temperature is 30 ℃, and incubation time is 20h; And the fermentation condition of bacillus is: the throughput of filtrated air is 1: 1, and mixing speed is 200 commentaries on classics/min, and cultivation temperature is 30 ℃, and incubation time is 40h, the most of gemma that forms of the little observation of microscope.After the fermentation ends, the thalline quantity of two genus reach 1,000,000,000/more than the ml, down absorption is air-dry and pulverize at 45 ℃ to use wheat bran and turfy soil (mass ratio 2: 8) respectively, mixes with 3: 7 ratio of mass ratio then, makes solid fungicide; Or respectively the zymocyte liquid of two genus is distinguished 1/2 of simmer down to original volume, and 3: 7 by volume ratio is mixed then, and liquid bacterial agent is made in aseptic packaging.
Embodiment 4
Gather the healthy soybean rhizosphere soil (Fuyuan County soybean field, Kiamusze City, Heilongjiang Province) in field, 10g soil adds the 90ml distilled water diluting and becomes bacteria suspension, utilize King ' B solid culture medium, ampicillin (the 40ug/ml that adds the degerming of 0.22um membrane filtration behind the medium sterilization, the sterile water dissolving), chloramphenicol (13ug/ml 95% dissolve with ethanol), cycloheximide (100ug/ml acetone solution), specificity screening pseudomonad (Pseudomonas spp.);
Bacteria suspension temperature in 80 ℃ of water-baths after the above-mentioned soil sample dilution is bathed 10min, utilizes beef extract-peptone solid culture medium screening bacillus (Bacillus spp.) then.
Gather field soybean root rot diseased plant root (Fuyuan County soybean field, Kiamusze City, Heilongjiang Province), the broken tissue in laboratory, (the sterilization back adds 2ml lactic acid with Martin's medium, suppressing bacterium and actinomycetic growth) the bacterial isolate fungi is mainly Fusarium oxysporum (Fusarium oxysporum), Fusarium graminearum (Fusarium graminearum) and Rhizoctonia solani Kuhn (RhizoctoniaSolani), and is stand-by.
Adopt the method for the dull and stereotyped Oxford of murphy juice (PDA) cup face-off, that is: with the disease fungus of above-mentioned screening respectively in inoculated and cultured on the PDA solid culture medium after 3 days, get the mycelia piece of diameter 8mm at the mycelia edge with the card punch of sterilizing, standby; Inoculate the mycelia piece respectively in new PDA solid plate central authorities, place Oxford cup (, placing again) in the position of distance mycelia piece 3cm, add the cultured products of each pseudomonad or bacillus in the cup of Oxford respectively on the flame about calcination 5s, cultivate after 3 days, detect the bacteriostasis rate of each bacterium.
Intend: the bacteriostasis rate of positive control is 100%, and the bacteriostasis rate of negative control is 0,
Filter out 36 pseudomonas and 21 bacillus with disease fungus activity that efficient inhibition screens, the mixed culture fermentation respectively of pseudomonad and bacillus, the condition of pseudomonad fermentation tank is: the throughput of filtrated air is 1: 1, mixing speed is 200 commentaries on classics/min, cultivation temperature is 30 ℃, and incubation time is 20h; And the fermentation condition of bacillus is: the throughput of filtrated air is 1: 1, and mixing speed is 200 commentaries on classics/min, and cultivation temperature is 30 ℃, and incubation time is 40h, and microscopically is observed most of gemma that forms.After the fermentation ends, the thalline quantity of two genus all reach 1,000,000,000/more than the ml, down absorption is air-dry and pulverize at 50 ℃ to use wheat bran and turfy soil (mass ratio 1: 9) respectively, mixes with 5: 5 ratio of mass ratio then, makes solid fungicide; Or respectively with 1/2 of the zymocyte liquid simmer down to original volume of two genus, 5: 5 by volume ratio is mixed then, and liquid bacterial agent is made in aseptic packaging.
Claims (9)
1. method for preparing the composite bacteria agent capable of preventing and treating root rot, it is characterized in that: the earth bacteria suspension fetches earth, utilize the method specificity screening pseudomonad (Pseudomonas spp.) and the bacillus (Bacillus spp.) that are coated with beef-protein medium behind King ' B medium and the 80 ℃ of water-bath 10min respectively, stand-by; Other gathers root rot diseased plant root, utilizes Martin's medium bacterial isolate fungi; Adopt the method for the dull and stereotyped Oxford of PDA cup face-off, bacterial strain with disease fungus activity that efficient inhibition screens, the composite composite bacteria agent capable that can obtain preventing and treating root rot after the fermentation process respectively will be sifted out in the pseudomonad of above-mentioned specificity screening and the bacillus again.
2. the method for preventing and treating the composite bacteria agent capable of root rot by the described preparation of claim 1, it is characterized in that: described King ' B nutrient media components is: show peptone 20g, potassium dihydrogen phosphate 1.2g, epsom salt 1.5g, glycerine 10ml, agar 15g, distilled water 1000ml, pH=7.4-7.6 mixes the back sterilization, the sterilization back adds ampicillin, chloramphenicol and the cycloheximide of 0.22um membrane filtration degerming.
3. the method for preventing and treating the composite bacteria agent capable of root rot by the described preparation of claim 1, it is characterized in that: described ampicillin is 40ug/ml with the sterile water concentration of ordinary dissolution; Chloramphenicol is 13ug/ml with 95% dissolve with ethanol concentration; Cycloheximide is 100ug/ml with acetone solution concentration.
4. the method for preventing and treating the composite bacteria agent capable of root rot by the described preparation of claim 1, it is characterized in that: described beef-protein medium component is: beef extract 5g, peptone 10g, sodium chloride 5g, agar 15g, distilled water 1000ml, pH=7.0-7.2.Soil supension temperature in 80 ℃ of water-baths was bathed 10 minutes, and dilution is coated with flat board again.
5. the method for preventing and treating the composite bacteria agent capable of root rot by the described preparation of claim 1, it is characterized in that: the prescription of described Martin's medium is: glucose 10g, peptone 5g, potassium dihydrogen phosphate 1g, magnesium sulfate 0.5g, 0.1% rose-bengal solution 3.3ml, agar 20g, distilled water 1000ml, the pH nature.The sterilization back adds 80% lactic acid 2ml.
6. the method for preventing and treating the composite bacteria agent capable of root rot by the described preparation of claim 1, it is characterized in that: solid PDA culture medium prescription used in the method for the dull and stereotyped face-off of described employing Oxford cup is as follows: potato peeling 200g, sucrose 20g, distilled water 1000ml, pH nature.Potato is cut into small pieces, and adds distilled water and boils 20min, filters, and filtrate adds sucrose, and constant volume adds 20g agar, 115 ℃ of autoclaving 30min to 1000ml.
7. the method for preventing and treating the composite bacteria agent capable of root rot by the described preparation of claim 1, it is characterized in that: the method for the dull and stereotyped Oxford of PDA cup face-off, disease fungus mycelia piece at the inoculation diameter 8mm of murphy juice solid plate central authorities, place the Oxford cup after the calcination on the flame in the position of distance mycelia piece 3cm, perhaps the mycelia piece was cultivated after 1 day, placed the Oxford cup again, added the cultured products of each stand-by pseudomonad or bacillus in the cup of Oxford, cultivate after 2-3 days, detect the bacteriostasis rate of each bacterium.
8. the method for preventing and treating the composite bacteria agent capable of root rot by the described preparation of claim 1, it is characterized in that: with pseudomonad through sifting out again and bacillus mixed culture fermentation respectively, the hybrid bacterial strain of pseudomonas is fermented and cultured in above-mentioned King ' the B medium of added with antibiotic not; The hybrid bacterial strain of bacillus carries out fermentation medium in beef-protein medium; The thalline quantity for the treatment of two genus all reach 1,000,000,000/more than the ml, use mass ratio 2: 8-1 respectively: absorption is air-dry and pulverize down at 40-50 ℃ for 9 wheat bran and turfy soil, and then with mass ratio 3: 7-5: 5 ratio is mixed, and makes solid fungicide; Or respectively with the zymocyte liquid of two genus all the simmer down to original volume 1/2, then by volume 3: 7-5: 5 ratio is mixed, and liquid bacterial agent is made in aseptic packaging.
9. preparation according to claim 1 prevents and treats the method for the composite bacteria agent capable of root rot, it is characterized in that: the condition of pseudomonad fermentation tank is; The throughput of filtrated air is 1: 0.8-1, and mixing speed is 180-200 commentaries on classics/min, and cultivation temperature is 28-30 ℃, and incubation time is 20-30h; And the fermentation condition of bacillus is: the throughput of filtrated air is 1: 0.8-1, and mixing speed is 180-200 commentaries on classics/min, and cultivation temperature is 28-30 ℃, and incubation time is 40-60h, helps forming gemma.
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| CN109362790A (en) * | 2018-12-10 | 2019-02-22 | 河北省农林科学院石家庄果树研究所 | It is a kind of for preventing and treating the complex microbial inoculum and preparation method thereof of pear tree root knot |
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| CN102839126B (en) * | 2012-09-21 | 2013-10-09 | 宋彦耕 | Microbial agent and application of antigravity and cropping effects of microbial agent |
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| CN105900653A (en) * | 2016-04-25 | 2016-08-31 | 江苏大学 | Green biological method for preventing and treating root rot disease of maize |
| CN109362790A (en) * | 2018-12-10 | 2019-02-22 | 河北省农林科学院石家庄果树研究所 | It is a kind of for preventing and treating the complex microbial inoculum and preparation method thereof of pear tree root knot |
| CN110607239A (en) * | 2019-07-16 | 2019-12-24 | 甘肃省科学院生物研究所 | Novel method for producing D-lactic acid by fermenting straws with sorangium japonicum and application |
| CN110607239B (en) * | 2019-07-16 | 2020-07-14 | 甘肃省科学院生物研究所 | Novel method for producing D-lactic acid by fermenting straws with sorangium japonicum and application |
| CN115097038A (en) * | 2022-06-22 | 2022-09-23 | 山东国仓健生物科技有限公司 | Screening and identifying method and application of soybean phytophthora root rot-resistant related metabolites |
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