CN111662837B - Bacillus atrophaeus and application thereof - Google Patents
Bacillus atrophaeus and application thereof Download PDFInfo
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Abstract
The invention discloses a strain of Bacillus atrophaeus and application thereof. The Bacillus atrophaeus provided by the invention is Bacillus atrophaeus WS-1, and the preservation number of the Bacillus atrophaeus WS-1 in the common microorganism center of China Committee for culture Collection of microorganisms is CGMCC No.16198. Experiments prove that the Bacillus atrophaeus WS-1 can inhibit streptomyces scabies and/or streptomyces acidocaldarius and/or streptomyces albidoflavus, and has important application value for preventing and treating potato scab. The invention has important application value.
Description
Technical Field
The invention belongs to the field of microorganisms, and particularly relates to bacillus atrophaeus and application thereof.
Background
The potato scab is a plant disease which damages potato tubers and mainly shows that suberized scab-shaped light brown scab spots or plaques which are nearly round to irregular appear on the surfaces of the tubers, and the hand feel is rough. Potato scab generally has two symptoms of reticulate scab and split scab. Under normal conditions, the reticulate disease spots and the split disease spots are only limited to the cortex, but the quality and the yield of the damaged potato blocks can be reduced, the damaged potato blocks are not storage-resistant, the appearance of the damaged potato blocks is not elegant, the commodity grade is greatly reduced, and certain economic loss is caused. In China, the incidence of diseases is low, the area of damage is small, and therefore people do not pay attention to the disease. However, in recent years, the disease incidence area in inner Mongolia, hebei, shandong and other areas is increasing, and the economic loss caused by the disease incidence area is also considerable. Therefore, scientific control of potato scab is urgently needed.
The existing research shows that the pathogenic bacteria causing potato scab are mainly streptomyces scab. Streptomyces scab belongs to conditional pathogenic bacteria, namely, the Streptomyces scab is easy to grow and cause potato diseases under the condition that the soil environment is slightly alkaline, but rarely occurs in the condition that the soil environment is slightly acidic.
At present, the prevention and treatment methods of potato scab mainly comprise the following steps: (1) selecting a seed potato without a disease; (2) applying organic fertilizer or green manure more times can partially inhibit diseases; (3) Performing crop rotation with Cucurbitaceae, leguminosae, and Liliaceae vegetables for more than 5 years; (4) Selecting a vegetable field with good water retention for planting, and watering in time when meeting drought in a potato bearing period; (5) chemoprevention: some chemical pesticides can be used for preventing and treating potato scab, but long-term use of chemical agents can cause pathogenic bacteria to easily generate drug resistance, so that the prevention and treatment effect is greatly reduced, and serious environmental pollution is easily caused to endanger the health of people and livestock; (6) biological control: the method screens microorganisms with good inhibition effect on pathogenic bacteria of potato scab and strong planting capability, and the microorganisms are applied in the potato production process and the farmland planting process to achieve the effect of inhibiting the growth of the pathogenic bacteria.
Disclosure of Invention
The invention aims to prevent and treat potato scab.
The invention firstly protects Bacillus atrophaeus (WS-1), the strain is preserved in China general microbiological culture Collection center (CGMCC for short, with the address of No. 3 of Xilu No.1 of Beijing Korean area north Cheng, no. 16198) in 2018 at 08-month and 02-month day, and the preservation number is CGMCC No.16198. Bacillus atrophaeus WS-1CGMCC No.16198 is abbreviated as Bacillus atrophaeus WS-1.
The invention also protects a microbial inoculum which can contain Bacillus atrophaeus WS-1.
The microbial inoculum can be used as at least one of the following a 1) to a 8): a1 ) inhibiting streptomyces scab; a2 Preparing a product for inhibiting streptomyces scabiosus; a3 ) inhibiting streptomyces scabiosus; a4 Preparing a product for inhibiting streptomyces scabiosus; a5 ) inhibiting streptomyces albidoflavus; a6 Preparing a product for inhibiting streptomyces albidoflavus; a7 ) inhibiting streptomyces which can cause potato scab; a8 ) to produce a product that inhibits streptomyces that can cause potato scab.
The invention also provides a preparation method of the microbial inoculum, which comprises the following steps: inoculating Bacillus atrophaeus WS-1 to a bacterial culture medium and culturing to obtain a bacterial liquid, namely the microbial inoculum.
The bacterial culture medium can be a potato liquid culture medium. The preparation method of the potato liquid culture medium specifically comprises the following steps: cutting peeled potato 200g into small pieces, adding 1.0L distilled water, and boiling for 30min; filtering with gauze, collecting filtrate, adding glucose 20.0g, KH2PO 4.0 g, mgSO4.7H2O 1.5g and vitamin B110 μ g into the filtrate, adjusting pH to 6.0, diluting with distilled water to volume of 1.0L, and sterilizing at 115 deg.C for 15min.
In the preparation method of the microbial inoculum, the specific culture conditions can be as follows: culturing at 30 + -2 deg.C and 150-250r/min (such as 150-200r/min, 200-250r/min, 150r/min, 200r/min or 250 r/min) under shaking for 1-3d (such as 1d, 2d or 3 d).
The microbial inoculum may include a carrier in addition to the active ingredient. The carrier may be a solid carrier or a liquid carrier. The solid carrier may be a mineral material, a plant material or a polymeric compound. The mineral material may be at least one of clay, talc, kaolin, montmorillonite, white carbon, zeolite, silica, and diatomaceous earth. The plant material may be at least one of corn flour, bean flour and starch. The high molecular compound may be polyvinyl alcohol and/or polyglycol. The liquid carrier can be an organic solvent, vegetable oil, mineral oil, or water. The organic solvent may be decane and/or dodecane. In the microbial inoculum, the active ingredient may be present in the form of cultured living cells, a fermentation broth of living cells, a filtrate of a cell culture, or a mixture of cells and a filtrate. The composition can be prepared into various dosage forms, such as liquid, emulsion, suspending agent, powder, granules, wettable powder or water dispersible granules.
According to the requirement, the microbial inoculum can also be added with a surfactant (such as Tween 20, tween 80 and the like), a binder, a stabilizer (such as an antioxidant), a pH regulator and the like.
The invention also protects the application of the Bacillus atrophaeus WS-1 or any one of the above bactericides, and the Bacillus atrophaeus WS-1 or any one of the above bactericides can be at least one of the following a 1) to a 8):
a1 ) inhibiting streptomyces scab;
a2 Preparing a product for inhibiting streptomyces scabiosus;
a3 ) inhibiting streptomyces scabiosus;
a4 Preparing a product for inhibiting streptomyces scabiosus;
a5 ) inhibiting streptomyces albidoflavus;
a6 Preparing a product for inhibiting streptomyces albidoflavus;
a7 ) inhibiting streptomyces which can cause potato scab;
a8 ) to produce a product that inhibits streptomyces that can cause potato scab.
In the above application, the product may be a bacteriostatic agent.
The application of the Bacillus atrophaeus WS-1 or any one of the microbial inoculum in preventing and treating diseases caused by the streptomyces scabies also belongs to the protection scope of the invention.
The application of the Bacillus atrophaeus WS-1 or any one of the microbial inoculum in preventing and treating diseases caused by the streptomyces scabies also belongs to the protection scope of the invention.
The application of the Bacillus atrophaeus WS-1 or any one of the microbial inoculum in preventing and treating diseases caused by streptomyces albidoflavus also belongs to the protection scope of the invention.
The application of the Bacillus atrophaeus WS-1 or any one of the microbial inoculum in preventing and treating diseases caused by streptomyces which can cause potato scab also belongs to the protection scope of the invention.
In the application, the streptomyces capable of causing potato scab can be specifically streptomyces scab, streptomyces acidocaldarius or streptomyces albidoflavus.
The application of Bacillus atrophaeus WS-1 or any one of the microbial inoculum in preventing and treating potato scab also belongs to the protection scope of the invention.
The invention also protects a product which contains the Bacillus atrophaeus WS-1 or any one of the microbial inoculum; the function of the product may be a 1) or a 3) or a 5) or a 7) or b 1) or b 2) or b 3) or b 4) or b 5):
a1 ) inhibiting streptomyces scab;
a3 ) inhibiting streptomyces scabiosus;
a5 ) inhibiting streptomyces albidoflavus;
a7 ) inhibiting streptomyces which can cause potato scab;
b1 Preventing and treating diseases caused by streptomyces scabies;
b2 Preventing and treating diseases caused by streptomyces acidoscabies;
b3 Preventing and treating diseases caused by streptomyces albidoflavus;
b4 Control of diseases caused by streptomyces which can cause potato scab;
b5 Preventing and treating potato scab.
The product may be a bacteriostatic agent.
In the above product, the "Streptomyces capable of causing potato scab" may be specifically Streptomyces scab, streptomyces acidocaldarius or Streptomyces albidoflavus.
Any one of the Streptomyces scabies may be specifically Streptomyces scabies (CGMCC) No.4.0076 (also called Streptomyces scabies (CGMCC) 4.76) or Streptomyces scabies (Streptomyces scabies) CGMCC No.4.1765 (also called Streptomyces scabies (CGMCC 4.1765).
Any one of the streptomyces scabies can be specifically streptomyces scabies (streptomyces acidiscabies) CGMCC No.4.1789.
Any one of the Streptomyces albidoflavus may specifically be Streptomyces albidoflavus (Streptomyces albicans) CGMCC No.4.3598.
In the above, the Streptomyces scabies (Streptomyces scabies) CGMCC No.4.0076, the Streptomyces scabies (Streptomyces scabies) CGMCC No.4.1765, the Streptomyces scabies (Streptomyces acissibicas) CGMCC No.4.1789 and the Streptomyces albidoflavus (Streptomyces albiflavus) CGMCC No.4.3598 are all stored in the China general microbiological culture collection management center (CGMCC, address: 100101 of institute of China academy of sciences, no. 3 of North West Lu 1 of south china, beijing city), the collection numbers are 4.76, 4.1765, 4.1789 and 4.3598 in sequence, and the public can obtain from the China general microbiological culture collection management center.
Experiments prove that the Bacillus atrophaeus WS-1 provided by the invention can inhibit streptomyces scabies and/or streptomyces acidocaldarius and/or streptomyces albidoflavus, and has important application value for preventing and treating potato scab.
Drawings
FIG. 1 shows the experimental results of step one in example 3.
FIG. 2 shows the results of the second step of example 3.
FIG. 3 shows the results of the third step in example 3.
FIG. 4 shows the results of the fourth experiment in example 3.
FIG. 5 shows the results of the fifth experiment in example 3.
FIG. 6 shows the results of the sixth step of example 3.
FIG. 7 shows the bacterial species of Streptomyces scabies (Streptomyces scabies) CGMCC No. 4.0076.
Deposit description
The strain name: bacillus atrophaeus
Latin name: bacillus atrophaeus
The strain number is as follows: WS-1
The preservation organization: china general microbiological culture Collection center
The preservation organization is abbreviated as: CGMCC
Address: xilu No.1 Hospital No. 3 of Beijing market facing Yang district
The preservation date is as follows: 08 and 02 months 2018
The registration number of the collection center: CGMCC No.16198
Detailed Description
The present invention is described in further detail below with reference to specific embodiments, which are given for the purpose of illustration only and are not intended to limit the scope of the invention.
The experimental procedures in the following examples are all conventional ones unless otherwise specified.
Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.
Streptomyces scabies (Streptomyces scabies) CGMCC No.4.0076 (also called Streptomyces scabies (Streptomyces scabies) CGMCC No. 4.76, the bacterial species are shown in figure 7), streptomyces scabies (Streptomyces scabies) CGMCC No.4.1765 (also called Streptomyces scabies (Streptomyces scabies) CGMCC No. 4.1765), streptomyces scabies (Streptomyces acidosis) CGMCC No.4.1789 and Streptomyces albidoflavus (Streptomyces albidoflavus) CGMCC No.4.3598 are all preserved in the common culture collection and management center of Chinese microorganisms (CGMCC, address: the microbial research institute of China academy of sciences, no. 3 of West road No.1 of the Korean-Yang area, beijing is 100101), the serial numbers of the strains are 4.76, 4.1765, 4.1789 and 4.3598 in sequence, and the public can obtain the Streptomyces scab (Streptomyces scab) CGMCC No.4.0076, 4.1765, 4.1789, 4.17698, 4.3598, 3598, respectively.
Potato liquid medium: cutting peeled potato 200g into small pieces, adding 1.0L distilled water, and boiling for 30min; filtering with gauze, collecting filtrate, adding glucose 20.0g, KH2PO 4.0 g, and MgSO4 4 .7H 2 1.5g of O and 10 mu g of vitamin B, adjusting the pH value to 6.0, adding distilled water to the volume of 1.0L, and sterilizing at 115 ℃ for 15min.
Potato solid medium: taking peeled potato 200g, cutting into small pieces, adding 1.0L distilled water, boiling for 30min; filtering with gauze, collecting filtrate, adding glucose 20.0g and KH 2 PO 4 3.0g, mgSO4.7H2O 1.5g, vitamin B1 μ g and agar 15.0g, adjusting pH to 6.0 and adding distilled water to volume of 1.0L, and sterilizing at 115 deg.C for 15min.
Potato solid plate: and (3) pouring 20mL of potato solid culture medium with the temperature of about 55 ℃ into a culture dish, and cooling to obtain the potato solid flat plate.
Example 1 isolation, identification and preservation of Bacillus atrophaeus (Bacillus atrophaeus) WS-1CGMCC No.16198
1. Separation of
1. Treatment of soil samples
(1) Adding 10g soil sample (soil for planting potato in Heiral area of Huroebel city, autonomous region of inner Mongolia) and appropriate amount of glass beads into 90mL sterile water, oscillating for 10min, standing for 30s, and making into soil stock solution (the dilution is 10 at this time) -1 ). The supernatant of 1mL of the soil stock solution was aspirated and added to a test tube containing 9mL of sterile water to mix well (the dilution was 10 at this time) -2 ) Then 1mL of the solution is sucked from the test tube and added into another test tube containing 9mL of sterile water to be uniformly mixed, and the like to prepare a diluent (the dilution is 10) -4 )。
2. Inoculating Streptomyces scabies 4.0076 to a potato solid plate, and standing and culturing at 28 deg.C for 4-5 days; then, 0.2mL of sterile water was added to the solid potato plate, and spores of Streptomyces scabies 4.0076 were scraped using sterile inoculation to obtain a suspension of pathogenic bacteria (the concentration of Streptomyces scabies 4.0076 was about 1.0X 10) 8 cfu/mL)。
3. Mixing 50 μ L of the diluent obtained in step 1 and 50 μ L of the pathogenic bacteria suspension obtained in step 2, uniformly coating on a potato solid plate, and standing and culturing at 28 deg.C for 2d. Bacterial colonies capable of forming inhibition zones are subjected to strain purification on a potato solid plate.
One of the screened bacteria was designated as bacterial WS-1.
2. Identification
1. Morphological identification
The bacteria WS-1 is in the shape of straight rod, has the size of 0.5-2.5 μm x (1.2-10) μm, and is usually arranged in pairs or chains with round ends. Upon juvenile culture, bacterial WS-1 staining was generally gram positive with a movement of periciliary flagella. The spores are oval and oval, and can resist adverse environment. Each bacterium produces a spore, and the spore formation is not inhibited by oxygen. Aerobic or facultative anaerobic. Has physiological properties of resisting adverse conditions such as pH, high osmosis, high salt content, etc. Chemolithotrophs. Of the fermentative or respiratory metabolic type. Generally catalase positive.
The bacterial WS-1 is inoculated to a potato solid medium and cultured at 28 ℃, and the morphology of a single colony is observed after 2 days. The results show that the bacterial WS-1 has convex and white colonies in the middle, irregular and dull surface folds and irregular edges.
2. 16S rRNA sequence homology analysis
The 16S rRNA of the bacteria WS-1 is shown as a sequence 1 in a sequence table.
By comparison, the bacterial WS-1 has the highest homology with Bacillus atrophaeus.
Thus, the bacterium WS-1 was identified as Bacillus atrophaeus.
3. Preservation of
The separated bacteria WS-1 in the first step are preserved in China general microbiological culture Collection center (CGMCC for short, no. 3 Siro 1 of Beijing market in the morning of the Yangxi district), and the preservation number is CGMCC No.16198, already in 2018, at the 02 th month of 08. The bacterial WS-1 is totally called Bacillus atrophaeus (Bacillus atrophaeus) WS-1CGMCC No.16198, which is abbreviated as Bacillus atrophaeus WS-1.
Example 2 expansion of Bacillus atrophaeus WS-1
The specific steps of the expanding propagation of the Bacillus atrophaeus WS-1 are as follows:
1. bacillus atrophaeus WS-1 was activated three times on potato solid medium. Inoculating 1-inoculated Bacillus atrophaeus WS-1 into a 250mL conical flask filled with 50mL of potato liquid culture medium, and performing shaking culture at 28 ℃ and 200r/min for 2d to obtain a bacterial liquid.
2. And (3) after the step 1 is finished, taking the bacterial liquid, centrifuging for 1min at 12000r/min, and collecting supernatant and precipitate.
3. After completion of step 2, the pellet was taken out and resuspended in 50mL of sterile water to obtain a cell suspension (the concentration of Bacillus atrophaeus WS-1 in the cell suspension was about 1.0X 10 10 cfu/mL)。
Example 3 bacteriostatic Effect of Bacillus atrophaeus WS-1
1. Inhibitory effect of Bacillus atrophaeus WS-1 on Streptomyces scabies 4.0076
The experiment was repeated three times and the mean value was taken. The specific steps of each experiment are as follows:
1. inoculating Streptomyces scabies 4.0076 to a potato solid plate, and standing and culturing at 28 deg.C for 4-5 days.
2. After step 1, the solid potato plate was taken, 0.2mL of sterile water was added, and then spores and mycelia on the surface of the solid potato plate were scraped off with a sterile inoculating spatula to obtain a pathogen suspension 1 (the concentration of streptomyces scabies 4.0076 was about 1.0 × 10) 8 cfu/mL)。
3. After step 2 was completed, 100 μ L of pathogen suspension 1 was taken and evenly spread on a potato solid plate.
4. And (3) after the step 3 is finished, uniformly dividing the potato solid plate into three equal parts, placing sterile filter paper with the diameter of 5mm in the center of each equal part, then respectively dotting and connecting 10 mu L of the bacterial liquid, the supernatant and the cell suspension obtained in the embodiment 2 on the sterile filter paper, standing and culturing for 3d at 28 ℃, and observing the bacteriostatic effect.
And (5) replacing the cell suspension in the step (4) with sterile water, and taking the cell suspension as a negative control without changing other steps.
The bacteriostatic effect of Bacillus atrophaeus WS-1 on Streptomyces scabies 4.0076 is shown in FIG. 1 (the left image is the front side of the solid potato plate, and the right image is the back side of the solid potato plate). The results showed that after 10. Mu.L of the bacterial solution, supernatant or cell suspension obtained in example 2 was spot-inoculated, a zone of inhibition appeared (the diameter of the zone of inhibition was about 13.3 mm); after the culture in the sterile water, no inhibition zone appears. Therefore, the Bacillus atrophaeus WS-1 has a certain inhibition effect on the streptomyces scabies 4.0076.
2. Inhibitory Effect of Bacillus atrophaeus WS-1 on Streptomyces scabies 4.1765
According to the method of the first step, the streptomyces scabies 4.0076 is replaced by the streptomyces scabies 4.1765, and the inhibition effect of the bacillus atrophaeus WS-1 on the streptomyces scabies 4.1765 is obtained when the other steps are not changed.
The bacteriostatic effect of Bacillus atrophaeus WS-1 on Streptomyces scabies 4.1765 is shown in FIG. 2 (left is front side of solid potato plate, right is back side of solid potato plate). The results showed that after 10. Mu.L of the bacterial suspension, supernatant or cell suspension obtained in example 2 was spot-inoculated, there was an inhibition zone (the diameter of the inhibition zone was about 14.0 mm); after the culture in the sterile water, no inhibition zone appears. Therefore, bacillus atrophaeus WS-1 has certain inhibition effect on Streptomyces scabies 4.1765.
3. Inhibitory Effect of Bacillus atrophaeus WS-1 on Streptomyces scabies 4.1789
Replacing the streptomyces scabies 4.0076 with the streptomyces scabies 4.1789 according to the method in the first step, and obtaining the inhibition effect of the bacillus atrophaeus WS-1 on the streptomyces scabies 4.1789 without changing other steps.
The bacteriostatic effect of Bacillus atrophaeus WS-1 on Streptomyces scabies 4.1789 is shown in FIG. 3 (left is front side of solid potato plate, right is back side of solid potato plate). The results showed that after 10. Mu.L of the bacterial suspension, supernatant or cell suspension obtained in example 2 was spot-inoculated, there was an inhibition zone (the diameter of the inhibition zone was about 13.6 mm); after the culture in the sterile water, no inhibition zone appears. Therefore, bacillus atrophaeus WS-1 has a certain inhibitory effect on Streptomyces scabies 4.1789.
4. Inhibitory Effect of Bacillus atrophaeus WS-1 on Streptomyces albidoflavus 4.3598
According to the method of the first step, the streptomyces scabies 4.0076 is replaced by the streptomyces albidoflavus 4.3598, and the other steps are not changed, so that the inhibition effect of the bacillus atrophaeus WS-1 on the streptomyces albidoflavus 4.3598 is obtained.
The bacteriostatic effect of Bacillus atrophaeus WS-1 on Streptomyces albidoflavus 4.3598 is shown in FIG. 4 (the left image is the front side of a potato solid plate, and the right image is the back side of the potato solid plate). The results showed that after 10. Mu.L of the bacterial suspension, supernatant or cell suspension obtained in example 2 was spot-inoculated, there was an inhibition zone (the diameter of the inhibition zone was about 13.8 mm); after the culture in the sterile water, no inhibition zone appears. Therefore, the Bacillus atrophaeus WS-1 has a certain inhibition effect on the streptomyces albidoflauvs 4.3598.
5. Mixed inhibition effect of Bacillus atrophaeus WS-1 on Streptomyces scabies 4.0076, streptomyces scabies 4.1765, streptomyces acidocaldarius 4.1789 and Streptomyces albidoflauvs 4.3598
The experiment was repeated three times and the mean value was taken. The specific steps of each experiment are as follows:
1. inoculating Streptomyces scabies 4.0076 to a potato solid plate, and standing and culturing at 28 deg.C for 4-5 days; then, 0.2mL of sterile water was added to the potato solid plate, and spores of Streptomyces scabies 4.0076 were scraped using sterile inoculation to obtain pathogen suspension 1 (the concentration of Streptomyces scabies 4.0076 was about 1.0X 10) 8 cfu/mL)。
2. Inoculating Streptomyces scabies 4.1765 to a potato solid plate, and standing at 28 deg.C for 4-5 days; then, 0.2mL of sterile water was added to the solid potato plate, and spores of Streptomyces scabies 4.1765 were scraped off using sterile inoculation to obtain pathogen suspension 2 (the concentration of Streptomyces scabies 4.1765 was about 1.0X 10) 8 cfu/mL)。
3. Inoculating Streptomyces scabies 4.1789 to a potato solid plate, and standing and culturing at 28 deg.C for 4-5 days; then, 0.2mL of sterile water was added to the potato solid plate, and the spores of Streptomyces scabies 4.1789 were scraped off with a sterile inoculating spatula to obtain a pathogen suspension 3 (the concentration of Streptomyces scabies 4.1789 was about 1.0X 10) 8 cfu/mL)。
4. Get4.3598 of streptomyces albidoflavus, inoculating the streptomyces albidoflavus into a potato solid plate, and standing and culturing for 4-5 days at 28 ℃; 0.2mL of sterile water was then added to the potato solid plate, and spores of Streptomyces albidoflauvs 4.3598 were scraped using a sterile inoculating spatula to obtain pathogen suspension 4 (concentration of Streptomyces albidoflauvs 4.3598 was about 1.0X 10 8 cfu/mL)。
5. And (5) after the steps 1 to 4 are finished, mixing the pathogenic bacteria suspension 1, the pathogenic bacteria suspension 2, the pathogenic bacteria suspension 3 and the pathogenic bacteria suspension 4 in equal volume to obtain a mixed bacteria suspension A.
6. After the step 5 is completed, 400 mu L of the mixed bacterial suspension A is uniformly coated on a potato solid plate.
7. And 6, taking the potato solid plate, placing sterile filter paper with the diameter of 5mm in the center of the potato solid plate, then dotting 10 mu L of the cell suspension obtained in the example 2 on the sterile filter paper, standing and culturing for 2d at 28 ℃, and observing the bacteriostatic effect.
The "cell suspension obtained in example 2" in step 7 was replaced with sterile water, and the other steps were not changed, and used as a negative control.
The bacteriostatic effect of Bacillus atrophaeus WS-1 on Streptomyces scabies 4.0076, streptomyces scabies 4.1765, streptomyces acidocaldarius 4.1789 and Streptomyces albidoflauvs 4.3598 is shown in FIG. 5. The results show that after the cell suspension obtained in example 2 is spot-jointed, inhibition zones (the diameter of the inhibition zone is about 13.3 mm) appear; after the culture in the sterile water, no inhibition zone appears. Therefore, the Bacillus atrophaeus WS-1 has certain inhibition effect on mixed bacteria consisting of streptomyces scabies 4.0076, streptomyces scabies 4.1765, streptomyces scabies acidocaldarius 4.1789 and streptomyces albidoflauvs 4.3598.
6. Co-culture bacteriostatic effect
1. The same as step 1 in the fifth step.
2. The same as step 2 in the fifth step.
3. The same as step five, step 3.
4. The same as step 4 in the fifth step.
5. After completion of steps 1 to 4, 100. Mu.L of pathogen suspension 1, 100. Mu.L of pathogen suspension 2, 100. Mu.L of pathogen suspension 3, 100. Mu.L of pathogen suspension 4 and 10. Mu.L of the cell suspension obtained in example 2 were mixed to obtain mixed bacterial suspension B.
6. And (5) after the step (5) is finished, uniformly coating the mixed bacterial suspension B on a potato solid plate. And (5) carrying out static culture at 28 ℃, and observing the bacteriostatic effect after culturing for 2d.
The co-culture bacteriostatic effect is shown in fig. 6. The results show that only Bacillus atrophaeus WS-1 grew during co-culture, whereas Streptomyces scabies 4.0076, streptomyces scabies 4.1765, streptomyces acidoscabies 4.1789 and Streptomyces albidoflauvs 4.3598 showed no signs of growth. Therefore, bacillus atrophaeus WS-1 has inhibitory effect on Streptomyces scabies 4.0076, streptomyces scabies 4.1765, streptomyces acidocaldarius 4.1789 and Streptomyces albidoflauvs 4.3598.
<110> institute of microbiology of China academy of sciences of Ningxia university
<120> Bacillus atrophaeus strain and application thereof
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 1413
<212> DNA
<213> Bacillus atrophaeus
<400> 1
catgcagtcg agcggacaga tgggagcttg ctccctgatg ttagcggcgg acgggtgagt 60
aacacgtggg taacctgcct gtaagactgg gataactccg ggaaaccggg gctaataccg 120
gatgcttgtt tgaaccgcat ggttcaaaca taaaaggtgg cttcggctac cacttacaga 180
tggacccgcg gcgcattagc tagttggtga ggtaatggct caccaaggca acgatgcgta 240
gccgacctga gagggtgatc ggccacactg ggactgagac acggcccaga ctcctacggg 300
aggcagcagt agggaatctt ccgcaatgga cgaaagtctg acggagcaac gccgcgtgag 360
tgatgaaggt tttcggatcg taaagctctg ttgttaggga agaacaagtg ccgttcaaat 420
agggcggcac cttgacggta cctaaccaga aagccacggc taactacgtg ccagcagccg 480
cggtaatacg taggtggcaa gcgttgtccg gaattattgg gcgtaaaggg ctcgcaggcg 540
gtttcttaag tctgatgtga aagcccccgg ctcaaccggg gagggtcatt ggaaactggg 600
gaacttgagt gcagaagagg agagtggaat tccacgtgta gcggtgaaat gcgtagagat 660
gtggaggaac accagtggcg aaggcgactc tctggtctgt aactgacgct gaggagcgaa 720
agcgtgggga gcgaacagga ttagataccc tggtagtcca cgccgtaaac gatgagtgct 780
aagtgttagg gggtttccgc cccttagtgc tgcagctaac gcattaagca ctccgcctgg 840
ggagtacggt cgcaagactg aaactcaaag gaattgacgg gggcccgcac aagcggtgga 900
gcatgtggtt taattcgaag caacgcgaag aaccttacca ggtcttgaca tcctctgaca 960
cccctagaga tagggcttcc ccttcggggg cagagtgaca ggtggtgcat ggttgtcgtc 1020
agctcgtgtc gtgagatgtt gggttaagtc ccgcaacgag cgcaaccctt gatcttagtt 1080
gccagcattc agttgggcac tctaaggtga ctgccggtga caaaccggag gaaggtgggg 1140
atgacgtcaa atcatcatgc cccttatgac ctgggctaca cacgtgctac aatggacaga 1200
acaaagggca gcgagaccgc gaggttaagc caatcccaca aatctgttct cagttcggat 1260
cgcagtctgc aactcgactg cgtgaagctg gaatcgctag taatcgcgga tcagcatgcc 1320
gcggtgaata cgttcccggg ccttgtacac accgcccgtc acaccacgag agtttgtaac 1380
acccgaagtc ggtgaggtaa cctttatgga gcc 1413
Claims (8)
1. Bacillus atrophaeus WS-1, the preservation number of which in China general microbiological culture Collection center is CGMCC No.16198.
2. A microbial preparation comprising Bacillus atrophaeus (WS-1CGMCC No. 16198) according to claim 1.
3. The method for preparing the microbial inoculum according to claim 2, which comprises the following steps: inoculating the Bacillus atrophaeus (Bacillus atrophaeus) WS-1CGMCC No.16198 of claim 1 to a bacterial culture medium and culturing to obtain a bacterial liquid, namely a microbial inoculum.
4. The use of the inoculant of Bacillus atrophaeus (WS-1CGMCC No. 16198) of claim 1 or claim 2, wherein the inoculant is at least one of the following a 1) to a 6):
a1 Inhibiting Streptomyces scabies;
a2 Preparing a product for inhibiting streptomyces scabiosus;
a3 ) inhibiting streptomyces scabiosus;
a4 Preparing a product for inhibiting streptomyces acidoscabicus;
a5 ) inhibiting streptomyces albidoflavus;
a6 To prepare a product for inhibiting streptomyces albidoflavus.
5. Use of the Bacillus atrophaeus (Bacillus atrophaeus) WS-1CGMCC No.16198 of claim 1 or the microbial inoculum of claim 2 in preventing and treating diseases caused by Streptomyces scabies.
6. The use of the Bacillus atrophaeus (Bacillus atrophaeus) WS-1CGMCC No.16198 or the microbial inoculum of claim 2 in preventing and treating diseases caused by Streptomyces scabies.
7. The use of the Bacillus atrophaeus (Bacillus atrophaeus) WS-1CGMCC No.16198 or the bacterial agent of claim 2 in preventing and treating diseases caused by Streptomyces albidoflavus.
8. A product containing the Bacillus atrophaeus (Bacillus atrophaeus) WS-1CGMCC No.16198 or the bacterial agent of claim 2.
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| CN112662585B (en) * | 2021-01-11 | 2022-04-29 | 中国科学院微生物研究所 | Bacillus atrophaeus DX-9 and application thereof |
| CN113201470B (en) * | 2021-03-23 | 2022-08-02 | 山东施可丰生物科技有限公司 | Carrier type soil remediation microbial inoculum |
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