CN112226387B - White bacillus and application thereof in preventing and treating basal rot of corn stalk - Google Patents
White bacillus and application thereof in preventing and treating basal rot of corn stalk Download PDFInfo
- Publication number
- CN112226387B CN112226387B CN202011128100.4A CN202011128100A CN112226387B CN 112226387 B CN112226387 B CN 112226387B CN 202011128100 A CN202011128100 A CN 202011128100A CN 112226387 B CN112226387 B CN 112226387B
- Authority
- CN
- China
- Prior art keywords
- corn
- bacillus
- cell suspension
- white
- rot
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
- C12N1/205—Bacterial isolates
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N63/00—Biocides, pest repellants or attractants, or plant growth regulators containing microorganisms, viruses, microbial fungi, animals or substances produced by, or obtained from, microorganisms, viruses, microbial fungi or animals, e.g. enzymes or fermentates
- A01N63/20—Bacteria; Substances produced thereby or obtained therefrom
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
Abstract
The invention discloses a strain of white bacillus and application thereof in preventing and treating basal rot of corn stalks. The bacillus albicans is bacillus albicans Leucobacter sp.M6560, and is preserved in the common microorganism center of China Committee for culture Collection of microorganisms, wherein the preservation number is CGMCC No.20598, and the preservation date is 9, month and 4 days 2020. M6560 has remarkable antagonistic effect on Fusarium moniliforme (Fusarium moniliforme) causing stalk rot disease of corn. Seed dressing before corn sowing and spraying of stem base and leaf surface after 4-5 leaves emerge are carried out by using the bacterial suspension of the strain, so that the occurrence of corn stem basal rot is remarkably reduced, and the prevention effect on the corn stem basal rot is 87.5%; and the corn can be planted in the rhizosphere and the phyllosphere for a long time after being used, so that the corn can be protected for a long time. The invention provides important strain resources for pollution-free prevention and treatment of corn diseases and has important practical value.
Description
Technical Field
The invention relates to a biocontrol bacterium and application thereof, in particular to a white bacillus and application thereof in preventing and treating basal rot of corn stalks.
Background
Corn stalk rot is an important soil-borne fungal disease that damages corn roots and stalk bases. The incidence rate is generally about 10%, and the serious incidence rate is more than 30%. China is harmful in places such as Guangxi, Hubei, Sichuan, Shandong, Shanxi, Shaanxi, Hebei, Liaoning, Jilin, Heilongjiang, Tianjin, Beijing and the like, and has become a large disease in the current corn production. Fungal type stalk rot is a kind of disease causing root system and stalk rot caused by single or compound infection of various pathogenic bacteria, and is mainly caused by Pythium aphanidermatum, Fusarium moniliforme, Colletotrichumgraminicola, carbon rot fungus macrophos phasecolina and the like. Fusarium causes stem rot which is common in the later growth stage of corn.
The main biocontrol bacteria currently used to control corn stalk rot caused by fusarium include: bacillus subtilis YNM-3CGMCC No.6933, Pseudomonas azotoformans WXCDD51 CGMCC No.12760, Bacillus (Bacillus sp.) WXCDD105 CGMCC No.12496, Bacillus subtilis HR15, Bacillus atrophaeus HR37 and the Bacillus belgii HR55 mixed flora, Bacillus methylotrophicus NJ13, Bacillus methylotrophicus TA-1CCTCC No. M2018362, Trichoderma harzianum LTR-2CGMCC No.1498, Bacillus megaterium JBN02, Bacillus methylotrophicus KNK-1 and Bacillus cereus BCB 01, Bacillus megaterium Bacillus thuringiensis Ajuss 14, Bacillus cereus Bsanensis 14, Bacillus cereus urea-producing Bacillus cereus 64, and the like. The strain resources for preventing the corn stalk rot caused by fusarium are very limited.
However, the prior art disclosed above has the disadvantages that the strain is difficult to fix the plant at the using part of the crop after the microbial inoculum is used, or the survival time after the plant is fixed is short, the abundance is low, the strain has a good disease prevention effect only in the short time after the microbial inoculum is used, and the lasting period is short. At present, the application of the bacillus albicans to the control of the corn stalk basal rot is not disclosed.
Disclosure of Invention
The invention provides a strain of white bacillus and application thereof in preventing and treating basal rot of corn stalks aiming at the defects. The Leuconobacter sp.M6560 strain provided by the invention can effectively prevent and treat basal rot of corn stalks. The strain and the cell suspension thereof have simple use method and easy operation, can be planted and survived for a long time after being applied to the root and the phyllosphere of the corn, keep higher abundance, form long-acting protection on the corn and have higher effect than chemical pesticide multi-bacteria. The biocontrol bacterium disclosed by the invention provides an important strain resource for green prevention and control of corn diseases, and has an important application value.
The technical scheme of the invention is that the white bacillus is Leucobacter sp.M6560, which is preserved in China general microbiological culture Collection center (CGMCC, address: No. 3 Hopkinson No.1 Hopkinson Kogyo Chen-Su-Tokyo, China academy of sciences) with the preservation number of CGMCC No.20598 and the preservation date of 2020, 9 and 4 days.
The invention also protects the application of the bacillus albicans in preventing and treating the corn stalk basal rot.
The corn stalk is rotten and is caused by Fusarium moniliforme.
A product for preventing and treating basal rot of corn stalk, the active component comprises cell suspension of the white bacillus Leucobacter sp.M6560 of claim 1, the effective viable count is 108-10CFU/mL。
The preparation method of the cell suspension of the white bacillus Leucobacter sp.M6560 comprises the following steps:
(1) inoculating the white bacillus soybean casein agar culture medium to a soybean casein culture medium, and culturing at 28-32 ℃ and 160-;
(2) transferring the culture to a soybean casein culture medium, culturing at 28-32 deg.C and 160-;
(3) collecting the culture solution, centrifuging at 8000rpm for 10-15min, discarding the supernatant, and weighing with sterile phosphate buffer solution containing 0.05% Tween-80Suspending white bacillus M6560 cells to obtain white bacillus M6560 with concentration of 108-1010CFU/mL cell suspension.
Preferably, the preparation method of the cell suspension of the Leucobacter sp.M6560 comprises the following steps:
(1) inoculating the white bacillus soybean casein agar culture medium to a soybean casein culture medium, and culturing at 30 ℃ and 180rpm for 12h to obtain a culture;
(2) transferring the culture to a soybean casein culture medium, and culturing at 30 ℃ and 180rpm for 72h to obtain a culture solution;
(3) the culture solution was collected and centrifuged at 6000rpm for 10min, the supernatant was discarded, and the cells of M6560, which is a white bacillus strain, were resuspended in sterile phosphate buffer containing Tween-80 at a mass concentration of 0.05% to obtain a cell suspension of M6560, which is a white bacillus strain.
A method of controlling basal rot of a corn stalk, comprising at least one of the following two steps:
(1) seed dressing before corn sowing by using the cell suspension of the Leucobacter sp.M6560 as the claim 1;
(2) use of a cell suspension of the Leuconobacter sp.M6560 strain according to claim 1 to spray the basal and foliar parts of shoots after emergence of 4-5 leaves.
The seed dressing specifically comprises the following steps: cell suspension diluted to 10 using Leucobacter sp7-109And (5) CFU/mL, soaking the corn seeds to be sown in the diluted cell suspension for 10-15min, taking out, draining until no suspension drips, and sowing.
The stem base and the leaf surface spraying after 4-5 leaves emerge specifically are as follows: cell suspension diluted to 10 using Leucobacter sp7-109CFU/mL, spraying the whole corn plant at the 4-5 leaf stage, and requiring the liquid to fully soak the corn plant during spraying.
The invention has the beneficial effects that:
the white bacillus strain M6560 used in the invention is separated from the intestinal tract of garlic maggot larvae, and a strong antibiotic environment is formed in the garlic maggot intestinal tract because garlic maggots eat garlic, so the strain has strong adaptability to adverse environments, can be planted and survived for a long time after being applied to the root and phyllosphere of corn, keeps high abundance, forms long-term protection on corn, and has higher effect than that of chemical pesticides.
The white bacillus M6560 strain provided by the invention can effectively prevent and control the corn stalk basal rot. The strain and the cell suspension thereof have simple use method and easy operation, can be planted and survived for a long time after being applied to the root and the phyllosphere of the corn, keep higher abundance, form long-acting protection on the corn and have higher effect than chemical pesticide multi-bacteria. The biocontrol bacterium disclosed by the invention provides an important strain resource for green prevention and control of corn diseases, and has an important application value.
Drawings
FIG. 1 shows the electrophoresis diagram of the PCR product of 16S rDNA of the strain M6560 of the present invention;
FIG. 2 shows the genetic evolutionary relationship analysis of 16S rDNA of the strain M6560 of the present invention;
FIG. 3 shows the antagonistic activity of the strain M6560 of the present invention against Fusarium moniliforme (Fusarium moniliforme); wherein the left side is strain M6560, and the right side is a control;
FIG. 4 shows the effect of potting experiments; wherein the left side is corn seedlings treated by M6560, and the right side is a control group inoculated with pathogenic bacteria;
FIG. 5 shows the long-term planting of the strain M6560 of the present invention in corn roots;
FIG. 6 shows the long-term planting of the strain M6560 of the present invention in the corn leaf space.
Detailed Description
For better understanding of the present invention, the technical solution of the present invention will be described in detail with specific examples, but the present invention is not limited thereto.
Example 1
Isolation and characterization
The strain isolation sample is obtained from larvae of Pistia virginiana in Vandah Garlic field of Taian, Shandong, and the collected larvae are placed in a sealed bag and recorded. The sealing bag is placed in an ice box for storage and is transported back to a laboratory for standby. After the larvae were brought back to the laboratory, the body surfaces were disinfected with 75% ethanol disinfectant for 15s, followed by rinsing with sterile water. The disinfection process is repeated three times. The disinfected larvae are placed under a dissecting mirror, heads and tails of the larvae are cut off by using an ophthalmic surgical scissors, then the larvae are dissected from the side surfaces of the larvae, and intestinal tracts are taken out. The intestinal tract was placed in a 1.5mL centrifuge tube, 100. mu.L phosphate buffer was added and the tube was ground thoroughly. Diluting to 10-8 times, coating the diluted solution on a soybean casein culture medium plate, culturing at 28 ℃ in an incubator for 24h, selecting a single colony, streaking the single colony on a new soybean casein culture medium plate, and selecting the streaked single colony and storing the single colony on a soybean casein culture medium agar plate.
Culturing the obtained pure strains by using a soybean casein culture medium for 12h, adding 25 mu L of the pure strains to one side of the edge of a PDA plate, which is 0.5cm away from the outer edge of the plate, then inoculating fusarium moniliforme cakes to one side of the opposite side of an inoculation point, using an isovolumetric liquid soybean casein culture medium instead of bacterial liquid as a contrast, setting 3 parallel controls for each bacterial strain, culturing for 5 days at 26 ℃ in the dark, and observing the growth condition and the bacteriostatic circle condition of fusarium moniliforme. Selecting bacterial strains with obvious inhibition on the growth of fusarium moniliforme for identification.
After the obtained strain is cultured for 12 hours by using a soybean casein culture medium, 1mL of bacterial liquid is taken, 2000g of the bacterial liquid is centrifuged for 10min to collect thalli, and the genomic DNA of the strain is extracted by using a DNA extraction kit. The method comprises the steps of adopting bacterial 16S rDNA universal primers 1492R (5'-GGCTCGAGCGGCCGCCCGGGTTACCTTGTTACGACTT-3') and 8F (5'-GCGGATCCGCGGCCGCTGCAGAGTTTGATCCTGGCTCAG-3') to amplify a bacterial 16S rDNA sequence by taking genome DNA as a template, sending a PCR product (shown in figure 1) after amplification to a Shanghai biological limited company for sequencing, uploading the sequence to a GenBank database to obtain a sequence number of MT808905, and when the GenBank database is used for identification, the similarity of the sequence of a strain M6560 and a Leucobacter genus is higher, but the sequence of the strain M6560 has a certain difference with a model strain (shown in figure 2), and finally the strain is identified as Leucobacter sp. The strain is preserved in China general microbiological culture Collection center, and the address is as follows: west road No.1, north chen, north china, beijing, chaoyang district; the preservation date is as follows: 9/4/2020, accession number: CGMCC No. 20598.
Example 2
Test for culturing white bacillus M6560 and Fusarium moniliforme in opposition
A60 mm petri dish was used to prepare a PDA plate for use, and a hole was punched at a position 0.5cm from the outer edge of the plate on the side of the edge of the PDA plate. Inoculating a small amount of plate culture of white bacillus M6560 to a soybean casein culture medium, culturing at 30 ℃ and 180rpm for 12h, taking 25 mu L of the plate culture, adding the 25 mu L of the plate culture into one side hole of the edge of a PDA plate, then inoculating fusarium moniliforme cake at one position on the opposite side of an inoculation point, using an isovolumetric liquid soybean casein culture medium to replace bacterial liquid as a contrast, setting 3 parallel controls for each bacterial strain, culturing at 26 ℃ for 5 days in the dark, and observing the growth condition and the inhibition zone condition of fusarium moniliforme. The results show that the white bacillus M0055 has a remarkable inhibitory effect on fusarium moniliforme (see figure 3).
Example 3
Experiment of potting
A small amount of white bacillus M6560 soybean casein agar culture is inoculated in 1mL soybean casein culture medium and cultured for 12h at 30 ℃ and 180 rpm. Transferring the culture into a triangular flask containing 150mL of soybean casein culture medium, and culturing at 30 ℃ and 180rpm for 72 h; 6 flasks of culture solution were collected and centrifuged at 6000rpm for 10min, the supernatant was discarded, and 100mL of sterile phosphate buffer containing 0.05% Tween-80 was used to resuspend the M6560 cells of Bacillus albus to obtain a mother solution of M6560 cell suspension of Bacillus albus. Subsequently, the cell suspension stock solution was further diluted to a concentration of 10 using the above-mentioned sterile phosphate buffer6、107、108、109、1010CFU/mL。
Using the dilution solution of the cell suspension of the bacillus albicans M6560 with different concentrations, soaking a group of 20 corn seeds in the diluted cell suspension for 10-15min, taking out, draining (until no suspension drips), and sowing. Another group of 20 corn seeds were soaked in sterile clear water for the above treatment, drained and sown. Transplanting the above seedlings with culture medium, wherein Fusarium moniliforme spore suspension (10 g) is added into the medium per 100g2CFU/mL)500mL of thoroughly wetted, potted with thoroughly wetted media and seeded with the two sets of treated corn seeds. Maize varietyZhengdan 958. Two groups of corns are placed in two trays for cultivation, so that cross contamination is avoided. The maize seedlings are cultured in a light incubator at the temperature of 30 ℃, the photoperiod L: D is 16:8, and the relative humidity is 50%. After 4-5 leaves of corn seedlings emerge, diluting liquid with different concentrations of a white bacillus M6560 cell suspension is used for spraying the whole corn plant until the corn plant is soaked by the liquid completely. After 10 days, the corn seedlings were observed for morbidity. The above experiment was repeated three times and the incidence was calculated. The results are shown in Table 1, the inoculation of the white bacillus M6560 obviously reduces the incidence rate of the corn stalk base rot in the potting experiment, and the incidence rate of the corn stalk base rot is gradually reduced with the increase of the application dosage, 108、109And 1010The effect is better under the concentration of CFU/mL.
TABLE 1 potted plant experimental results
Example 4 application of Bacillus albus M6560 in prevention and treatment of basal rot of maize stalk
The field test was conducted in Mazhuang, Taian, Shandong province, 5-8 months in 2020. The corn variety is Zhengdan 958.
A small amount of white bacillus M6560 soybean casein agar culture is inoculated in 1mL soybean casein culture medium and cultured for 12h at 30 ℃ and 180 rpm. Transferring the culture into a triangular flask containing 150mL of soybean casein culture medium, and culturing at 30 ℃ and 180rpm for 72 h; collecting culture solution, centrifuging at 6000rpm for 10min, discarding supernatant, and resuspending Bacillus albicans M6560 cells with 100mL sterile phosphate buffer containing 0.05% Tween-80 to obtain Bacillus albicans M6560 with concentration of 109CFU/mL cell suspension.
The test was divided into three groups of treatments. First group used the above 10 in corn planting9CFU/mL of the white bacillus M6560 cell suspension for seed dressing treatment. 109The CFU/mL cell suspension was used at 500mL per kilogram corn seed. And adding the cell suspension into the seeds, fully stirring and uniformly mixing, airing and sowing. After 4-5 leaves of corn seedling, use 109CFU/mL of Bacillus albus M6560 cell suspension 1000 times dilution, whole plant spray, when spraying, require liquid fully soak corn plant. The second group was treated with seed dressing and post-emergence spraying as above using sterilized water instead of the suspension of M6560 cells of Bacillus albicans as a control group. The third group uses carbendazim water solution (the effective dose of carbendazim is 10mg/mL) to replace the white bacillus M6560 cell suspension diluent in the first group for seed dressing treatment; spraying after-seedling with carbendazim water solution (effective dose of carbendazim is 10 mg/L). Each group was divided into 5 cells for testing. The disease index is investigated during the silking period of corn.
The grading standard of the degree of the corn stalk base rot disease is as follows: level 0: no symptoms, green leaves; level 1: 1-2 water-stain-shaped disease spots appear on leaf sheaths, diseased stems do not become soft, and withered leaves account for less than 1/4 of the whole plant; and 3, level: the base of the plant stem is discolored, the plant stem is soft when pinched by hands, and the withered leaves account for 1/4-1/2 of the whole plant; and 5, stage: the stem of the plant is obviously softened, the fruit cluster drops, and the green dead leaves account for 1/2-2/3 of the whole plant; and 7, stage: the stem of the plant is soft, even the stem is broken and the plant is lodging, and the green and withered leaves account for more than 2/3 of the whole plant.
Disease index ∑ (number of disease strains at each stage × disease grade value)/(number of survey total strains × highest grade value) × 100 control effect [ (] control group disease index-experimental group disease index)/control group disease index × 100
The results show that the disease index of the white bacillus M6560 treatment group is 0.62 percent, and the control effect is as high as 63.09 percent; the disease index of the carbendazim treatment group is 0.92%, and the control effect is 45.23%. The disease indexes of the two groups are obviously reduced compared with those of a clear water control group, and the prevention effect of the bacillus albicans M6560 on the corn stalk basal rot caused by fusarium moniliforme is better than that of carbendazim.
TABLE 2 results of the field test
Values in this column followed by different letters represent significant differences (one-way ANOVA, P <0.05)
Example 5
Planting condition of bacillus albicans M6560 in corn root and leaf space
The field test was conducted in Mazhuang, Taian, Shandong province, 5-8 months in 2020. The corn variety is Zhengdan 958.
A small amount of white bacillus M6560 soybean casein agar culture is inoculated in 1mL soybean casein culture medium and cultured for 12h at 30 ℃ and 180 rpm. Transferring the culture into a triangular flask containing 150mL of soybean casein culture medium, and culturing at 30 ℃ and 180rpm for 72 h; collecting culture solution, centrifuging at 6000rpm for 10min, discarding supernatant, and resuspending Bacillus albicans M6560 cells with 100mL sterile phosphate buffer containing 0.05% Tween-80 to obtain Bacillus albicans M6560 with concentration of 109CFU/mL cell suspension. Use of the above-mentioned 10 in corn seeding9CFU/mL of the white bacillus M6560 cell suspension for seed dressing treatment. 109The CFU/mL cell suspension was used at 500mL per kilogram corn seed. And adding the cell suspension into the seeds, fully stirring and uniformly mixing, airing and sowing. After 4-5 leaves of corn seedling, use 109CFU/mL of a 1000-fold dilution of a white bacillus M6560 cell suspension is sprayed on the whole strain until the corn plant is soaked by the liquid completely.
Corn root samples were collected at 0 (untreated), 10, 20, 30, 40, and 50 days after emergence of seedlings, and after 1g of the root samples were used to thoroughly clean the surface, DNA was extracted with a cleaning solution for future use. Corn heart and leaf samples are collected at 0 (untreated), 10, 20, 30, 40 and 50 days of post-emergence spraying respectively, 50g of the heart and leaf samples are used for fully cleaning the surface, and then DNA is extracted by cleaning liquid for standby. The use concentration is 109CFU/mL of M6560 LB medium for white bacilli 1mL extracted DNA, followed by 10-fold gradient dilution (10 times) with sterile water-1-10-9) And performing a fluorescent quantitative PCR amplification experiment by using a specific primer of the bacillus albicans M6560 to establish a standard curve of a fluorescence value and the number of cells. Subsequently, the planting situation of M6560 in the cucumber rhizosphere is determined by a standard curve. The results showed that on day 10 after treatment with M6560 suspension of Albacterium, white rodsThe planting quantity of the bacteria M6560 in the rhizosphere reaches 1.1 multiplied by 10 per gram of root7CFU, the number of the white bacilli M6560 planted in the rhizosphere is stabilized at 10 per gram of root within 50 days after the white bacilli M6560 floating solution treatment5CFU is of order magnitude, and can be planted on the root of the corn for a long time (figure 5 in the attached drawing of the specification). Similarly, within 50 days after spraying with the white bacillus M6560 suspension, the white bacillus can be planted in heart leaves for 103CFU/50g, can be planted in the phyllosphere for a long time (figure 6 in the attached figure of the specification).
Claims (8)
1. White bacillus (B)Leucobactersp.) M6560, wherein the white bacillus M6560 is preserved in the China general microbiological culture Collection center with the preservation number of CGMCC No.20598, the preservation date of 2020, 9 and 4 days, and the preservation address of No. 3 Hospital No.1 North Chen West Lu of the Yang ward area in Beijing.
2. The use of the white bacillus M6560 in the control of corn stalk rot in claim 1, characterized in that the corn stalk rot is made of Fusarium moniliforme (Fusarium moniliforme) (R)Fusarium moniliforme) And (4) causing.
3. A product for controlling basal rot of corn stalks, characterized in that the active ingredient comprises a cell suspension of the white bacillus M6560 of claim 1, the effective viable count is 108-10 CFU/mL。
4. The product for controlling corn stalk rot according to claim 3, wherein the cell suspension of the white bacillus M6560 is prepared by the following method:
(1) inoculating the white bacillus M6560 soybean casein agar culture medium into a soybean casein culture medium, culturing at 28-32 deg.C and 160-;
(2) transferring the culture to a soybean casein culture medium, culturing at 28-32 deg.C and 160-;
(3) collecting culture solution, centrifuging at 8000rpm for 10-15min, and discardingSupernatant, resuspending the M6560 cells of Bacillus albus with sterile phosphate buffer containing 0.05% Tween-80 to obtain M6560 cell of Bacillus albus with concentration of 108-1010CFU/mL cell suspension.
5. The product for controlling corn stalk rot according to claim 4, wherein the cell suspension of the white bacillus M6560 is prepared by the following method:
(1) inoculating white bacillus M6560 soybean casein agar culture medium to soybean casein culture medium, and culturing at 30 deg.C and 180rpm for 12 hr to obtain culture;
(2) transferring the culture to a soybean casein culture medium, and culturing at 30 ℃ and 180rpm for 72h to obtain a culture solution;
(3) the culture solution was collected and centrifuged at 6000rpm for 10min, the supernatant was discarded, and the cells of M6560, which is a white bacillus strain, were resuspended in sterile phosphate buffer containing Tween-80 at a mass concentration of 0.05% to obtain a cell suspension of M6560, which is a white bacillus strain.
6. A method for preventing and controlling corn stalk basal rot caused by fusarium moniliforme is characterized by comprising the following two steps:
(1) seed dressing before corn sowing by using the cell suspension of the white bacillus M6560 as the claim 1;
(2) spraying the cell suspension of the white bacillus M6560 of claim 1 on the base of the stem and the leaf surface of 4-5 emerged leaves.
7. The method for controlling the basal rot of corn stalks caused by fusarium moniliforme according to claim 6, wherein the seed dressing is specifically: cell suspension diluted to 10 using M6560 white bacillus7-109And (5) CFU/mL, soaking the corn seeds to be sown in the diluted cell suspension for 10-15min, taking out, draining until no suspension drips, and sowing.
8. The pest control cluster of claim 6The method for treating the corn stalk base rot caused by fusarium graminearum is characterized in that the spraying on the base of the stalk and the leaf surface after 4-5 leaves emerge is specifically as follows: cell suspension diluted to 10 using M6560 white bacillus7-109 CFU/mL, spraying the whole corn plant at the 4-5 leaf stage, and requiring the liquid to fully soak the corn plant during spraying.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011128100.4A CN112226387B (en) | 2020-10-20 | 2020-10-20 | White bacillus and application thereof in preventing and treating basal rot of corn stalk |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011128100.4A CN112226387B (en) | 2020-10-20 | 2020-10-20 | White bacillus and application thereof in preventing and treating basal rot of corn stalk |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112226387A CN112226387A (en) | 2021-01-15 |
| CN112226387B true CN112226387B (en) | 2022-02-15 |
Family
ID=74117438
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011128100.4A Active CN112226387B (en) | 2020-10-20 | 2020-10-20 | White bacillus and application thereof in preventing and treating basal rot of corn stalk |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112226387B (en) |
Family Cites Families (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2015092549A2 (en) * | 2013-12-18 | 2015-06-25 | Dupont Nutrition Biosciences Aps | Biologicals for plants |
| RU2624054C2 (en) * | 2015-12-07 | 2017-06-30 | Общество С Ограниченной Ответственностью "Арктический Научно-Проектный Центр Шельфовых Разработок" | Strain leucobacter aridicollis arc 6 vkpm as-1992 destructor of oil and oil products |
| CN110157641B (en) * | 2019-05-20 | 2023-05-05 | 黑龙江省农业科学院土壤肥料与环境资源研究所 | Biocontrol bacterium BV23 for preventing and treating corn stem-based rot and application thereof |
| CN110452840B (en) * | 2019-07-26 | 2020-10-30 | 中南大学 | White bacillus and application thereof in reduction of hexavalent chromium |
-
2020
- 2020-10-20 CN CN202011128100.4A patent/CN112226387B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN112226387A (en) | 2021-01-15 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN111172080B (en) | Bacillus belgii and application thereof | |
| JP4883814B2 (en) | Microorganisms having ability to control plant diseases, and plant disease control agents using the microorganisms | |
| KR101569737B1 (en) | Novel endophytic bacteria Bacillus oryzicola isolated from rice rhizosphere and development of a natural biopesticide and plant strengthener using same | |
| CN112175888B (en) | Bacillus belgii Hsg1949 and application thereof | |
| CN108034601B (en) | Bacillus amyloliquefaciens and application thereof | |
| CN110452832B (en) | Acid-resistant bacillus amyloliquefaciens Kc-5 and application thereof | |
| KR101346698B1 (en) | Endophytic Bacillus methylotrophicus EML-CAP7 with antimicrobial activity against plant pathogens and MRSA | |
| CN114501999A (en) | Biopesticide and biofertilizer composition | |
| CN112251375A (en) | Haemobacter and application thereof in prevention and treatment of cucumber fusarium wilt | |
| CN107075459B (en) | Novel bacterium belonging to the genus Bacillus and use thereof | |
| CN117165494A (en) | Kiwi fruit canker biocontrol strain Wq-1 and application thereof | |
| KR20080045346A (en) | Bacillus subtilis m27 and biological control of sclerotinia rot by using the same | |
| KR100746310B1 (en) | Effects of a soil-born Paenibacillus polymyxa strain KPB3 on suppression of bacterial wilt disease caused by Ralstonia solanacearum | |
| CN109609403B (en) | Biocontrol bacterium and application thereof in prevention and control of downy mildew of crops | |
| CN108384736B (en) | Bacterium with inhibiting effect on citrus sorangium cellulosum and screening method thereof | |
| CN116240126A (en) | Multifunctional bacillus belgium SB10 and application thereof | |
| CN113151079B (en) | Paenibacillus polymyxa KDB and application thereof | |
| CN114085788B (en) | Pseudomonas amazonensis strain and application thereof | |
| CN112226387B (en) | White bacillus and application thereof in preventing and treating basal rot of corn stalk | |
| KR20110120748A (en) | Bacillus amyloliquefaciens cp1 and contro method of strawberry anthracnose using the same | |
| CN113416679A (en) | Bacillus methylotrophicus, microbial inoculum comprising bacillus methylotrophicus and application of bacillus methylotrophicus | |
| CN115851476A (en) | Rice root endophytic bacillus altitudinis 258R-7 and biological agent and application thereof | |
| Zhang et al. | Decline in Cucumis melo L. Caused by Monosporascus cannonballus | |
| KR20130134875A (en) | Culture medium for preventing disease and promoting growth of plant, manufacturing method of microbial agent using the same | |
| KR100403896B1 (en) | Antagonistic bacillus ehimensis against vegetables damping-off |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |