CN116515714A - Bacillus subtilis Bacillus subtilis T S7B 6b and application thereof - Google Patents
Bacillus subtilis Bacillus subtilis T S7B 6b and application thereof Download PDFInfo
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- CN116515714A CN116515714A CN202310668749.2A CN202310668749A CN116515714A CN 116515714 A CN116515714 A CN 116515714A CN 202310668749 A CN202310668749 A CN 202310668749A CN 116515714 A CN116515714 A CN 116515714A
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- 244000063299 Bacillus subtilis Species 0.000 title claims abstract description 18
- 241000221662 Sclerotinia Species 0.000 claims abstract description 14
- 238000004321 preservation Methods 0.000 claims abstract description 8
- 230000002401 inhibitory effect Effects 0.000 claims abstract description 6
- 238000009629 microbiological culture Methods 0.000 claims abstract description 5
- 238000011282 treatment Methods 0.000 claims description 15
- 244000052616 bacterial pathogen Species 0.000 claims description 12
- 241000221696 Sclerotinia sclerotiorum Species 0.000 claims description 11
- 230000002265 prevention Effects 0.000 claims description 7
- 235000014469 Bacillus subtilis Nutrition 0.000 claims description 6
- 244000005700 microbiome Species 0.000 claims description 3
- 238000000855 fermentation Methods 0.000 claims description 2
- 230000004151 fermentation Effects 0.000 claims description 2
- 230000000813 microbial effect Effects 0.000 claims description 2
- 230000000694 effects Effects 0.000 abstract description 17
- 241001558929 Sclerotium <basidiomycota> Species 0.000 abstract description 7
- 230000003405 preventing effect Effects 0.000 abstract description 5
- 230000015572 biosynthetic process Effects 0.000 abstract description 2
- 230000000443 biocontrol Effects 0.000 description 19
- 201000010099 disease Diseases 0.000 description 19
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 19
- 241000894006 Bacteria Species 0.000 description 14
- 238000011081 inoculation Methods 0.000 description 10
- 241000219198 Brassica Species 0.000 description 9
- 235000003351 Brassica cretica Nutrition 0.000 description 9
- 235000003343 Brassica rupestris Nutrition 0.000 description 9
- QKSKPIVNLNLAAV-UHFFFAOYSA-N bis(2-chloroethyl) sulfide Chemical compound ClCCSCCCl QKSKPIVNLNLAAV-UHFFFAOYSA-N 0.000 description 9
- 235000010460 mustard Nutrition 0.000 description 9
- 238000012360 testing method Methods 0.000 description 9
- 241000196324 Embryophyta Species 0.000 description 7
- 239000002068 microbial inoculum Substances 0.000 description 6
- 239000007788 liquid Substances 0.000 description 5
- 235000021110 pickles Nutrition 0.000 description 5
- 240000006394 Sorghum bicolor Species 0.000 description 4
- 235000011684 Sorghum saccharatum Nutrition 0.000 description 4
- 235000013339 cereals Nutrition 0.000 description 4
- 230000005764 inhibitory process Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 239000000843 powder Substances 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 240000007124 Brassica oleracea Species 0.000 description 3
- 235000003899 Brassica oleracea var acephala Nutrition 0.000 description 3
- 235000011301 Brassica oleracea var capitata Nutrition 0.000 description 3
- 235000001169 Brassica oleracea var oleracea Nutrition 0.000 description 3
- 238000012258 culturing Methods 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 230000001613 neoplastic effect Effects 0.000 description 3
- 238000005507 spraying Methods 0.000 description 3
- 241000499436 Brassica rapa subsp. pekinensis Species 0.000 description 2
- 241000219193 Brassicaceae Species 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000002689 soil Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 235000010149 Brassica rapa subsp chinensis Nutrition 0.000 description 1
- 235000000536 Brassica rapa subsp pekinensis Nutrition 0.000 description 1
- 235000016623 Fragaria vesca Nutrition 0.000 description 1
- 240000009088 Fragaria x ananassa Species 0.000 description 1
- 235000011363 Fragaria x ananassa Nutrition 0.000 description 1
- 241000233866 Fungi Species 0.000 description 1
- 241000638919 Kleinia stapeliiformis Species 0.000 description 1
- 208000031888 Mycoses Diseases 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 244000131316 Panax pseudoginseng Species 0.000 description 1
- 235000003181 Panax pseudoginseng Nutrition 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 208000006278 hypochromic anemia Diseases 0.000 description 1
- 238000011835 investigation Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 239000000575 pesticide Substances 0.000 description 1
- 239000000447 pesticide residue Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Classifications
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- 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
-
- 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
- A01N63/22—Bacillus
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
- A01P3/00—Fungicides
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- 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
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- 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
- C12R2001/07—Bacillus
- C12R2001/125—Bacillus subtilis ; Hay bacillus; Grass bacillus
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- Chemical & Material Sciences (AREA)
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- Biotechnology (AREA)
- Organic Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Virology (AREA)
- Biochemistry (AREA)
- Plant Pathology (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Medicinal Chemistry (AREA)
- Environmental Sciences (AREA)
- Pest Control & Pesticides (AREA)
- Biomedical Technology (AREA)
- Dentistry (AREA)
- Agronomy & Crop Science (AREA)
- Mycology (AREA)
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Abstract
The invention discloses a bacillus subtilis Bacillus subtilis T S6b and application thereof, wherein the strain is preserved in China general microbiological culture Collection center (China general microbiological culture Collection center) for 12 months and 25 days in 2022, the preservation address is North Chen Xiyu No. 1, 3 in the Korean region of Beijing, and the preservation number is CGMCC No:26223. the invention has the advantages of inhibiting the growth of sclerotinia hyphae and the formation of sclerotium, having relatively obvious inhibiting effect on sclerotinia and good preventing and treating effect on stem nodule sclerotinia.
Description
Technical Field
The invention belongs to the technical field of biological strain application, and particularly relates to bacillus subtilis Bacillus subtilis T S6b and application thereof.
Background
The sclerotinia rot of the cruciferae is a worldwide fungal disease caused by sclerotinia sclerotiorum, wherein the sclerotinia sclerotiorum and the sclerotinia sclerotiorum (also called hot pickled mustard tuber) are important diseases in the current production, and the sclerotinia sclerotiorum is sclerotinia sclerotiorum Sclerotinia sclerotiorum, and the sclerotinia sclerotiorum can be developed in the whole growth period of the cruciferae crops. The damage symptoms of cabbage and hot pickled mustard tuber are similar, the leaves, stems and stems of the damaged plants in the seedling stage grow into water-stain-shaped spots, then the spots are gradually enlarged, the tissues are wet-rotted and white hyphae grow on the spots, and the whole plant is withered and rotten in the later stage; the adult plant period can be the positions of the leaves, the petioles, the tumor stems and the fruit pods of cruciferous crops, the diseased part is in a water-immersed round disease spot at the initial stage of disease occurrence of the disease plant, and then the disease part is spread to the whole plant, so that the whole plant is rotted, white hyphae grow on the diseased part, the white hyphae gradually grow into round or oval particles along with the time, and black brown granular sclerotium is formed on the rotted plant at the later stage. The most main means for preventing and treating sclerotinia is still to apply chemical pesticides at present, and the method has the characteristics of high prevention and treatment cost, large dosage, difficult control of pesticide residues in the field and unfriendly ecological environment, so that the search for beneficial microorganisms for efficiently preventing and treating sclerotinia is a problem which needs to be solved at present.
Disclosure of Invention
The invention aims to solve the problems and provide bacillus subtilis Bacillus subtilis T S6b and application thereof.
In order to achieve the above purpose, the present invention provides the following technical solutions:
the invention comprises bacillus subtilis Bacillus subtilis T S7 b and application thereof.
The strain is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) for 12 months and 25 days in 2022, the preservation address is 1 # 3 of North Chen Xiyu in the Chaoyang area of Beijing city, and the preservation number is CGMCC No:26223.
a microbial agent comprising the bacillus subtilis T7S6b.
An agent for inhibiting sclerotinia rot of a cruciferous crop, comprising a fermentation broth of said bacillus subtilis T7S6b.
Use of bacillus subtilis Bacillus subtilis T S6b in the prevention or treatment of sclerotinia rot of a cruciferous crop.
Further, the pathogenic bacteria of the sclerotinia rot of the cruciferous crop is sclerotinia sclerotiorum Sclerotinia sclerotiorum.
To sum up:
the invention has the advantages of inhibiting the growth of sclerotinia hyphae and the formation of sclerotium, having relatively obvious inhibiting effect on sclerotinia and good preventing and treating effect on stem nodule sclerotinia.
Drawings
FIG. 1 is a graph showing a plate-confrontation test of Bacillus subtilis Bacillus subtilisT S6b, wherein A: CK; b: T7S6b.
FIG. 2 is a schematic diagram of an in-vitro inoculation prevention test of bacillus subtilis Bacillus subtilis T S6b provided by the embodiment of the invention;
FIG. 3 is a graph showing comparison of control effects after spraying of the bacillus subtilis Bacillus subtilis T S6b biocontrol bacteria powder in the embodiment of the invention;
Detailed Description
The following examples are for illustrative purposes only and are not intended to limit the scope of the invention.
Bacterial strain origin
The example of the invention employs a soil dilution method to isolate the T7S6b strain from strawberry rhizosphere soil in the Fuling mountain of Chongqing. The T7S6b with good screening and preventing effects is sent to China center for type identification (CICC), and the final identification result is shown as bacillus subtilis (Bacillus subtilis). The strain is preserved in China general microbiological culture Collection center (CGMCC) in the 12 th month 28 of 2022, and the preservation number is CGMCC NO.26223.
Biocontrol effect
1. Materials and methods
1.1 Screening of biocontrol bacteria
The center of the PDA plate is connected with a pathogenic bacteria cake, 4 sterile oxford cups are placed at 4 corners 20mm away from the bacteria cake, 50 μl of bacteria liquid to be detected is respectively added, an equal amount of sterile NA culture liquid is added as a reference, the bacteria liquid is cultured for 3-5d in a 30 ℃ incubator, and the width of a bacteria inhibition zone and the diameter of a pathogenic bacteria colony are measured.
1.2 indoor control Effect determination
The seedling stage agar block inoculation method is adopted. Inoculating pathogenic bacteria on PDA plate, culturing for 3d, and collecting mycelium pellet with colony edge perforation diameter of 5 mm. And (3) in-vitro inoculation: respectively inoculating pathogenic mycelium blocks to leaves of hot pickled mustard tuber, cabbage and Chinese cabbage plants, wherein each leaf is inoculated with 1 mycelium block; at the same time, the concentration is 1.0 to 3.0X10 10 CFU/ml biocontrol bacteria liquid is sprayed around the inoculation part, and after the biocontrol bacteria liquid is subjected to moisture preservation and culture for 5-7 days, the disease condition is investigated. The disease classification is slightly changed according to the classification level of the severity of the sclerotium disease of the rape. Each treatment was inoculated 5 times and repeated 3 times.
1.3 field control effect determination
Adopts a sorghum grain inoculation method. Inoculating pathogenic bacteria on a PDA culture medium, culturing for 3-5d in a constant temperature incubator at 28 ℃, taking mycelium circular blocks with the diameter of 5mm punched at the edge of a colony, inoculating into triangular flasks filled with sterilized half-cooked sorghum grains, inoculating 3 mycelium blocks into each flask, and culturing for 5-7 d in the constant temperature incubator at 28 ℃. The sorghum grains with bacteria are sowed on the leaf of the preserved szechuan pickle, each strain of preserved szechuan pickle is inoculated at 3 positions, and the disease condition of the preserved szechuan pickle is investigated when the preserved szechuan pickle is harvested. Each treatment was inoculated with 60 strains and repeated 3 times. The disease classification refers to slight change of the classification level of the severity of the root rot of the pseudo-ginseng in the field.
Taking the biocontrol strain T7S6b as a research object, preparing the biocontrol strain T7S6b into biocontrol bacteria powder, and inoculating bacteria-carrying sorghum grains before sealing the forests of the hot pickled mustard tuber. The test was designed with 3 study factors of T7S6b biocontrol bacteria powder application time point, application times and application concentration (see Table 1), 1 for another CK treatment, 9 treatments, and 3 replicates for each treatment.
Table 1 field efficacy test treatment conditions
2. Results
2.1 Biocontrol bacterium screening
1 strain T7S6b with relatively obvious inhibition effect on sclerotinia sclerotiorum S1 (Sclerotinia sclerotiorum) is obtained by screening about 3000 strains, and the relative inhibition rate can reach 95% as shown in the figure 1 of the indoor flat plate opposite test result.
2.2 indoor control Effect determination
The control effect test results of indoor in-vitro inoculation of cruciferous crops such as hot pickled mustard tuber, cabbage and Chinese cabbage leaves are shown in figure 2, and after the leaves are sprayed on the strain T7S6b, the pathogenic bacteria S1 are basically not infected or infected and are not spread, so that the T7S6b has a good inhibition effect on the pathogenic bacteria.
2.3 field control effect determination
The field control effect test results show that all test treatments have a certain control effect on the sclerotium disease of the hot pickled mustard tuber, after the T7S6b biocontrol microbial inoculum is sprayed, the boundaries of the leaf and neoplastic disease spots are subjected to chlorosis, and later stage gradual withering is carried out, so that the leaf and neoplastic disease spots fall off, as shown in figure 3, the spreading speed of the leaf and neoplastic disease spots of the hot pickled mustard tuber is effectively reduced, and the control on the sclerotium disease of the hot pickled mustard tuber is realized. Wherein the T7S6b biocontrol microbial inoculum with the concentration of 300 g/mu is sprayed for 7d after the inoculation of the pathogenic bacteria and 14d after the inoculation of the pathogenic bacteria in the treatment 7, the control efficiency is up to 71.04 percent, the yield per mu is up to 3210kg, the yield is increased by 6.23 percent, and the income per mu is increased by 226 yuan according to 1.20 yuan/kg.
The prevention and treatment effect of the biocontrol microbial inoculum sprayed after inoculation of pathogenic bacteria is better than that before inoculation of pathogenic bacteria, which further proves that the T7S6b biocontrol microbial inoculum has better treatment effect on the sclerotium disease of the hot pickled mustard tuber. In addition, after the biocontrol microbial inoculum is sprayed, the yield of the preserved szechuan pickle can be obviously increased, and the leaves of the preserved szechuan pickle plants in the field are combined to show a state of evergreen and not withering, so that the T7S6b biocontrol microbial inoculum has a certain growth promoting effect.
Table 2 investigation results of field control test
Application of biocontrol strain T7S6b
In conclusion, a set of application technical schemes with better prevention and treatment effects on stem nodule diseases of stem nodule diseases are integrated preliminarily, namely, biocontrol bacteria are sprayed for 1 time after the stem nodule is swelled (in the middle and late twelve months), biocontrol bacteria are sprayed for 1 time before harvesting (in the upper one month), and the spraying concentration is 10 9 cfu/ml, 300g of fungus powder per mu is added with water for spraying, and the prevention and treatment effect on the stem nodule disease is best.
The foregoing description of the preferred embodiments of the invention is not intended to limit the invention to the precise form disclosed, and any such modifications, equivalents, and alternatives falling within the spirit and scope of the invention are intended to be included within the scope of the invention.
Claims (5)
1. The bacillus subtilis Bacillus subtilis T S6b is characterized in that the strain is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) in the 12 th month of 2022, the preservation address is the number 3 of North Chen Xiyu No. 1 in the Korean region of Beijing, and the preservation number is CGMCC No:26223.
2. a microbial agent comprising the Bacillus subtilis T7S6b according to claim 1.
3. An agent for inhibiting sclerotinia rot of a crucifer crop, comprising the fermentation broth of bacillus subtilis T7S6b according to claim 1.
4. Use of bacillus subtilis Bacillus subtilis T S6b in the prevention or treatment of sclerotinia sclerotiorum in cruciferous crops.
5. The use according to claim 4, wherein the pathogenic bacteria of sclerotinia of crucifer crops is sclerotinia sclerotiorum Sclerotinia sclerotiorum.
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CN202310668749.2A CN116515714A (en) | 2023-06-07 | 2023-06-07 | Bacillus subtilis Bacillus subtilis T S7B 6b and application thereof |
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CN202310668749.2A CN116515714A (en) | 2023-06-07 | 2023-06-07 | Bacillus subtilis Bacillus subtilis T S7B 6b and application thereof |
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CN202310668749.2A Pending CN116515714A (en) | 2023-06-07 | 2023-06-07 | Bacillus subtilis Bacillus subtilis T S7B 6b and application thereof |
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN116004466A (en) * | 2023-01-07 | 2023-04-25 | 内蒙古农业大学 | Bacterial strain for preventing and treating sclerotinia rot of sunflower, screening method and application |
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2023
- 2023-06-07 CN CN202310668749.2A patent/CN116515714A/en active Pending
Cited By (2)
Publication number | Priority date | Publication date | Assignee | Title |
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CN116004466A (en) * | 2023-01-07 | 2023-04-25 | 内蒙古农业大学 | Bacterial strain for preventing and treating sclerotinia rot of sunflower, screening method and application |
CN116004466B (en) * | 2023-01-07 | 2024-06-04 | 内蒙古农业大学 | Bacterial strain for preventing and treating sclerotinia rot of sunflower, screening method and application |
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