CN114854618A - Bacillus belgii SF327 and application thereof - Google Patents
Bacillus belgii SF327 and application thereof Download PDFInfo
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- CN114854618A CN114854618A CN202210198705.3A CN202210198705A CN114854618A CN 114854618 A CN114854618 A CN 114854618A CN 202210198705 A CN202210198705 A CN 202210198705A CN 114854618 A CN114854618 A CN 114854618A
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- oryzae
- bacillus belgii
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- 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
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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
- C12N1/02—Separating microorganisms from their culture media
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
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- 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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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
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Abstract
The invention relates to a Bacillus belgii SF327 strain and application thereof. The Bacillus belgii SF327 strain is preserved in China center for type culture Collection (CCTCC M2022130) at 18 months 2 in 2022. The Bacillus bellisianus has obvious growth promoting effect on green vegetables, can generate indole-3-acetic auxin, has obvious inhibiting effect on xanthomonas oryzae xanthophyllum blight and alternaria streak germ, and has broad-spectrum antibacterial activity on plant pathogenic fungi such as rice blast, cucumber fusarium wilt, phytophthora capsici, colletotrichum gloeosporioides, rubber tree tip spore colletotrichum anthracnose and the like. The results of the invention show that the strain has application prospects of disease prevention and growth promotion, provides good biocontrol resources for the research and development of growth-promoting microbial agents for green vegetables, the prevention and the treatment of rice bacterial leaf blight, rice streak and rice blast, and lays a foundation for the research and the development of novel microbial agents for rubber tree anthracnose.
Description
Technical Field
The invention belongs to the field of application of microbial technology, and particularly relates to a bacillus beiLeisi SF327 strain and application thereof.
Background
Cruciferae (Cruciferae) has a total of 375 genera, about 3200 species, and is widely distributed around the world. 95 genera, 425 varieties, 124 varieties, 7 introduction genera and 20 varieties (Leming, Jia Jianguang, and the like. identification of host range of plasmodiophora brassicae Cruciferae.2011) are distributed in China. The brassica and radish in cruciferae plants are main vegetable and oil crops in China, or are used as spicy seasonings, some types of the brassica and radish are important medicinal plants (such as isatis), some types of the brassica and radish are ornamental plants (such as violet), some types of the brassica and radish are honey-source plants, and the brassica and radish are important economic crops.
Leptoma brassicae is a pathogenic bacterium with serious harm on cruciferous plants, and the resulting clubroot disease is called plant cancer. Plasmodiophora brassicae mainly infects roots of host crops (Dekhuijzen H.M. the accucure of free and bound cytokines in clubroots and Plasmodiophora brassica. induced turn over tissue cultures, physiologia plantarum,1980,49(2): 169) 176.), crops can be infected in all growth and development periods, and the earlier the infection occurs, the greater the damage to the plants, the more serious the incidence (Zhang forest, Dihang. cruciferous vegetables Plasmodiophora occurrence and control measures, Shanghai vegetables, 2014(4): 48-49.). The leaves at the base of the diseased plants become yellow and wilted, and the whole plants die when the disease is serious, thereby causing great economic loss. In the late stage of disease, the roots are rotted, and the dormant spores are released into the soil, and the dormant spores can generally survive in the soil for more than 20 years, which is extremely threatening to continuous cropping farmland (Dixon G. Plasmodiophora fibres in environment. journal of Plant Growth Regulation,2009b,28(3): 212-. 228.). If the eradication by rotation takes more than 3 years, the feasibility is low, so the clubroot happens once in the planting field, and the effect is serious and long-term. The long-term use of common chemical prevention and control measures can not only improve the drug resistance of pathogenic bacteria, but also bring negative effects such as soil degradation and the like (Zhang Niang Bo. Chengdu Cruciferae clubroot investigation and medicament prevention and control screening. Sichuan agriculture university 2010.).
In recent years, more and more researches are carried out on plant rhizosphere growth-promoting bacteria (PGPR), the application of the PGPR can replace the use of chemical fertilizers to a certain extent, and the soil environment of farming is improved (Zhang Wantong, Lichao, and the like. the research on the effect of the plant rhizosphere growth-promoting bacteria fertilizer in a high cold meadow for replacing the chemical fertilizers. the grassland science report, 2021, (29)7: 1423-. For example, Dunaliella salina growth-promoting bacteria can improve the tolerance of pakchoi to salt stress (Ruihong, xuwenxin, etc. Dunaliella salina growth-promoting bacteria have an influence on the salt tolerance of pakchoi. Shanxi university report of agriculture (Nature science edition), 2021, 41 (5): 112. clan 120.), some can improve the agronomic characters of agricultural products while improving the growth amount of crops (Huangshu Chao, Houwan, etc. three strains of growth-promoting bacteria and the influence of mixed microbial agents thereof on the growth and quality of asparagus lettuce. Zhejiang agriculture report, 2021, 33(7): 1212. clan 1221.), some can further improve the rhizosphere microecology of crops while relieving the injury of salt and alkali stress on the crops (Wang Yu, Liushuang, etc.. 3 strains of salt and alkali-tolerant growth-promoting bacteria have an influence on the root microecology of mung bean. drought agriculture research in 2022, 40(1): 139. clan 145.).
The comprehensive development and utilization of growth-promoting bacteria are important research points in recent years, the development of growth-promoting bacteria resources not only can play a role in immediate effect, but also can reduce the use of pesticides and improve the ecological conditions of Soil, and is an important mode for promoting the development of ecological agriculture and the recovery of ecological environment (Liu S L, Maimaitiaili B, et al.
Disclosure of Invention
The invention aims to provide a bacillus beiLeisi SF327 and application thereof.
The Bacillus belgii has a growth promoting effect on the green vegetables, and has a remarkable antagonistic effect on rice bacterial blight, rice streak disease, rice blast, cucumber fusarium wilt, phytophthora capsici, colletotrichum gloeosporioides and rubber tree tip spore colletotrichum gloeosporioides. The Bacillus beleisi can provide help for growth promotion of green vegetables, prevention and control of rice diseases and prevention and control of rubber tree anthracnose diseases.
The purpose of the invention can be realized by the following technical scheme:
the first aspect of the present invention is: a strain of Bacillus belgii is provided, is separated from soil of garden estuary of Haikun City of Haikun province in 2018, 11 and 8 days, and is named as Bacillus belgii (Bacillus velezensis) SF327, and the Bacillus belgii is preserved in China Center for Type Culture Collection (CCTCC) in 2022 and 18 days, with the preservation number of CCTCC M2022130.
Performing plate culture on a solid culture medium, and observing that the colony color of the Bacillus belgii is milky white, the edge is not smooth and irregular, and the surface is dry, rough and opaque; gram staining results presented positive results.
Through physiological and biochemical tests, the bacillus belgii is a gram-positive bacterium, short rod-shaped, spore-producing, aerobic or facultative anaerobic bacterium.
The Bacillus belgii is capable of secreting 3-hydroxybutanone to produce acetomethylmethanol and gelatinase; can not hydrolyze o-nitrobenzene-galactoside, arginine, lysine, ornithine, sodium citrate, sodium thiosulfate and the like; glucose, mannitol, rhamnose, and the like cannot be oxidized; can produce acid by utilizing 22 carbon sources such as glycerol, L-arabinose, ribose, D-xylose, glucose, fructose, mannose and the like.
The Bacillus belgii has growth promoting effect on Brassica oleracea.
The growth promoting effect comprises that the overground fresh weight of the small green vegetables can be obviously increased by applying the Bacillus belgii, and the Bacillus belgii can produce auxin such as Indole-3-acetic acid (IAA).
The Bacillus belgii has antagonistic effect on Xanthomonas oryzae.
The Xanthomonas oryzae includes two pathogenic varieties of Xanthomonas oryzae (Xanthomonas oryzae pv. oryzae, Xoo) and Xanthomonas oryzae (Xanthomonas oryzae pv. oryzae, Xoc).
The antagonistic action of Bacillus belgii SF327 on rice bacterial blight and rice bacterial streak disease is detected by a plate confronting culture method, and the antagonistic activity of Bacillus belgii SF327 on 2 types of pathogenic bacteria is proved to be obvious.
The second aspect of the present invention is: providing a method for isolating said bacillus belgii: the method comprises the steps of adding sterile water into a soil sample by adopting a gradient dilution plate coating method, dissolving and oscillating for 20min, sequentially diluting the soil sample into soil bacterium suspensions with different gradients, respectively coating the soil bacterium suspensions on an NA (NA) plate inoculated with rice streak disease germs, observing the formation of an inhibition zone, scribing bacterial colonies capable of generating an obvious inhibition zone on the NA plate to obtain single bacterial colonies, carrying out PCR (polymerase chain reaction) amplification by utilizing a 16S rRNA (ribonucleic acid) gene of bacteria, constructing a phylogenetic tree, and determining the classification status of the bacterial strains.
The 16S rRNA gene of Bacillus belgii SF327 is shown in SEQ ID NO. 1.
The invention uses 16S rRNA gene to construct phylogenetic tree, carries out the comparative analysis of genetic relationship, and combines the physiological and biochemical characteristics of the strain to prove that the strain is Bacillus velezensis.
The third aspect of the present invention is: the application of the Bacillus belgii is provided, which comprises the following applications:
in a specific embodiment of the invention, the bacillus beijerinckii SF327 has a significant antagonistic effect on both rice bacterial blight and rice streak disease.
In one embodiment of the present invention, the rice bacterial blight comprises PXO99 A The rice streak disease germ comprises RS 105.
In one embodiment of the present invention, the Bacillus belgii SF327 has antagonistic action against a variety of plant pathogenic fungi, including, in particular, Magnaporthe oryzae (Magnaporthe oryzae), Fusarium oxysporum f.sp.cummerinum, Phytophthora capsici (Phytophthora capsici), Colletotrichum gloeosporioides (Colletotrichum gloeosporioides), and Colletotrichum trichothecoides (Colletotrichum acutum).
In one embodiment of the invention, the bacillus beijerinckii SF327 has a growth promoting effect on brassica chinensis, and particularly comprises the effect of obviously increasing the overground fresh weight of brassica chinensis.
In one embodiment of the invention, the growth promoting effect on Brassica oleracea comprises the ability of Bacillus belgii to produce auxins of the class 1H-Indole-3-acetic acid (IAA).
In a specific embodiment of the present invention, the bacillus belgii is used as a brassica oleracea growth promoting agent, and at the same time, the bacillus belgii is used as a Xanthomonas oryzae (Xanthomonas oryzae) antagonist, and the use of the bacillus belgii is used as a plant pathogenic fungi antagonist.
Compared with the prior art, the Bacillus belezii SF327 provided by the invention belongs to plant growth-promoting rhizobacteria (PGPR), has a remarkable growth-promoting effect on vegetables, can generate indole-3-acetic acid auxin, has a remarkable inhibiting effect on xanthomonas oryzae and alternaria streak germ, and also has broad-spectrum antibacterial activity on plant pathogenic fungi such as rice blast, fusarium oxysporum, phytophthora capsici, colletotrichum hevea, colletotrichum gloeosporum and the like.
The results of the invention show that the strain has broad-spectrum antibacterial and growth-promoting capabilities, shows that the strain has application prospects of disease prevention and growth promotion, provides good biocontrol resources for research and development of small vegetable growth-promoting bactericides and prevention and treatment of bacterial blight, streak disease and rice blast of rice, and lays a foundation for research and development of novel microbial agents for rubber tree anthracnose. Meanwhile, the biological control potential of the strain is to be further developed, and a foundation is laid for the research and development of multifunctional biological control inoculants.
Drawings
FIG. 1 is a photograph of Bacillus belgii SF327 under a microscope (1000X) and shows the colony morphology.
FIG. 2 shows the alignment of the gene sequences of Bacillus belgii SF 32716S rRNA and a Neighbor-Joining phylogenetic tree constructed using Virgibacillus pantothecicus NCDO 1765(X60627) as an outgrowth.
FIG. 3.16 gel electrophoresis results of S rRNA gene. Wherein 1 represents a 2k Marker; 2 represents the product of the 16S rRNA gene.
FIG. 4 is a graph showing the effect of Bacillus beleisi SF327 on the production of indole-3-acetate auxin.
FIG. 5 is a graph showing the effect of Bacillus belgii SF327 on growth promotion of Brassica oleracea.
FIG. 6 shows the Bacillus belgii SF327 pairs against rice bacterial blight strain PXO99 A The antagonistic effect of (2).
FIG. 7 is a graph showing the antagonistic effect of Bacillus belgii SF327 against rice streak disease germ RS 105.
FIG. 8 is a graph showing the antagonistic effect of Bacillus belgii SF327 against Pyricularia oryzae.
FIG. 9 is a graph showing the antagonistic effect of Bacillus belgii SF327 against cucumber fusarium oxysporum.
FIG. 10 is a graph showing the antagonistic effect of Bacillus beleisis SF327 against Phytophthora capsici.
FIG. 11 is a graph showing the antagonistic effect of Bacillus beleisis SF327 on colletotrichum gloeosporioides.
FIG. 12 is a graph showing the antagonistic effect of Bacillus beleisii SF327 on colletotrichum gloeosporioides.
Detailed Description
The invention is further illustrated by the following examples, which are not intended to limit the scope of the invention. The experimental methods without specifying specific conditions in the following examples were selected according to the conventional methods and conditions, or according to the commercial instructions. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not to be construed as limiting the invention.
The strain media used in the following examples are as follows:
NA solid Medium (g/L): 3g of beef extract, 5g of polypeptone, 10g of cane sugar, 1g of yeast powder and 15g of agar powder, dissolving the mixture in water, adjusting the pH value to 7.0-7.2, and sterilizing the mixture under high pressure (121 ℃ for 20 min).
NB liquid medium (g/L): 3g of beef extract, 5g of polypeptone, 10g of sucrose and 1g of yeast powder, adding water to dissolve the beef extract, diluting to 1000mL, adjusting the pH value to 7.0-7.2, and sterilizing under high pressure (121 ℃,20 min).
PDA solid medium (g/L): 200g of potato, 20g of glucose and 15g of agar are dissolved in water, and finally the volume is determined to be 1000mL, the pH value is 7.0-7.2, and the mixture is sterilized by high pressure (121 ℃,20 min).
Auxin assay YM Medium (g/L): m (g/L): mannitol 5g, K 2 HPO 4 0.25g, NaCl 0.05g, yeast extract 1.5g, tryptophan 0.05g, adding water to dissolve and fix the volume to 1000mL, adjusting pH to 7.0, and autoclaving (121 ℃,20 min).
Example 1 preparation of Bacillus belgii SF327
1. Source of soil
Soil of garden of Haidian estuary of Haikou city of Hainan province collected in 11 and 8 days of 2018
2. Screening of strains
(1) Soil sample collection
Collecting soil samples of about 10cm around the surface layer of the root of a fruit tree plant, and collecting 200g of soil samples at each point. 3 soil samples are collected in each land, and the sampling time, place and type are recorded. And (4) storing the collected soil sample at normal temperature for separating bacteria.
(2) Isolation of bacteria
Plate dilution method: weighing 10g of soil sample into a conical flask, adding 90mL of sterile water, oscillating in a shaking table at 200rpm and 28 ℃, taking out after 20min, and standing at room temperature for 10min to prepare a soil bacterium suspension stock solution. The stock solution was subjected to gradient dilution to obtain 10 each 0 、10 -1 、10 -2 、10 -3 、10 -4 、10 -5 Total 6 dilutions. Respectively taking 200 μ L of the bacterial suspension diluent, and uniformly coating on RS105 (OD) containing rice streak disease bacteria 600 2.0) were performed in 3 replicates per gradient. The plate was placed in an incubator at 28 ℃ for 48 hours and observed.
(3) Bacterial purification
Observing and selecting single bacterial colony with obvious bacteriostatic circle, streaking and purifying on NA plate, culturing in 28 deg.C incubator, 12 hr later selecting single bacterial colony, and numbering in sequence.
(4) Preservation of bacteria
Inoculating a single bacterial colony of the strain in an NB liquid culture medium, culturing for 12h in a shaking table at 28 ℃ and 180rpm, sucking 1mL of bacterial liquid and 1mL of 50% sterile glycerol, slightly shaking and uniformly mixing, and storing at-80 ℃ for a long time.
(5) Screening of growth-promoting bacteria
Auxin assay:
screening and content determination of the strain secreting the auxin IAA by a spectrophotometer method. The prepared modified YM medium was injected into a test tube and sterilized. Each tube was inoculated with the test strain under sterile conditions. Culturing at 135r/min for 96h in a shaker at 28 deg.C. The supernatant was retained by centrifugation, mixed with the colorimetric solution and incubated at room temperature for 30min, whereupon a pink color appeared, indicating IAA production. And measuring the OD value at the wavelength of 530nm, substituting the OD value into an IAA standard curve, and calculating the IAA content of the endophytes.
FIG. 4 shows the effect of Bacillus beleisi SF327 on indole-3-acetic acid (IAA) production.
Fresh weight determination method of small green vegetables:
the aerial parts of the small vegetables were picked (the underground parts of the vegetables were removed with scissors) and weighed immediately, and the fresh weight of the aerial parts was calculated.
The effect of Bacillus belgii SF327 on growth promotion of Brassica oleracea is shown in FIG. 5.
Finally, selecting Bacillus velezensis (SF 327) which has been preserved in China Center for Type Culture Collection (CCTCC) at 18/2 in 2022, with the preservation number of CCTCC M2022130.
FIG. 1 shows the microscopic (1000X) observation photograph and colony morphology of Bacillus belgii SF 327.
Example 2 identification of the 16S rRNA Gene of Bacillus beilesiensis SF327
The genome DNA of the strain SF327 was extracted by the CTAB method using Shanghai Senno organism GmbH, and the gene primers of the bacterial 16S rRNA: 27F 5'-AGAGTTTGATCCTGGCTCAG-3' and 1492R 5'-TACGGCTACCTTGTTACGACTT-3', using the extracted genomic DNA as a template, and performing PCR amplification to obtain the desired fragment. The PCR reaction system is as follows: the system is finished according to the actual situation.
TABLE 1 Ex-Taq polymerase chain reaction System
The basic conditions of the PCR reaction are as follows: pre-denaturation at 95 ℃ for 3min, denaturation at 95 ℃ for 30s, annealing at 65 ℃ for 30s, extension at 72 ℃ for 30s, pre-extension at 72 ℃ for 10min, and storage at 4 ℃ for 35 cycles. After the reaction is finished, the PCR products are checked by 1% agarose gel electrophoresis, a gel imager is used for detecting and recording the result, the gel electrophoresis result of the 16S rRNA gene is shown in figure 3, and 1 in figure 3 represents Marker; 2 represents the product of the 16S rRNA gene. The PCR stock was sent to Shitanhua Biotech (Shanghai) Co., Ltd for sequencing. The sequencing result is analyzed by using DNA Star and BLAST comparison is carried out on NCBI website to determine the species of the related strains.
The 16SrRNA gene of Bacillus belgii SF327 is shown in SEQ ID NO. 1.
The sequence alignment result of Bacillus belgii SF 32716S rRNA gene is shown in FIG. 2, and the result shows that the Neighbor-Joining phylogenetic tree constructed by using Virgibacillus pantothenticacus NCDO 1765(X60627) as an outer branch: the 16S rRNA gene of SF327 has 99.86% similarity with Bacillus velezensis.
Example 3 physiological and Biochemical characterization of Bacillus belgii SF327
The bacillus beleisi SF327 has the physiological and biochemical characteristics that: can secrete 3-hydroxy butanone to produce acetyl methyl methanol and gelatinase; can not hydrolyze o-nitrobenzene-galactoside, arginine, lysine, ornithine, sodium citrate, sodium thiosulfate and the like; glucose, mannitol, rhamnose, and the like cannot be oxidized; can produce acid by utilizing 22 carbon sources such as glycerol, L-arabinose, ribose, D-xylose, glucose, fructose, mannose and the like. See tables 2 and 3.
TABLE 2 physiological and biochemical characteristics of Strain SF 327-enzyme Activity, carbon Source Oxidation
+: positive reaction; -: negative reaction;
TABLE 3 physiological and biochemical characteristics of Strain SF 327-acid production Using carbon sources
+: positive reaction; -: negative reaction; w-weakly positive reaction
Example 4 determination of antagonistic Spectrum by Bacillus belgii SF327
1) Determination of antagonistic activity of Bacillus beilesiensis SF327 on Xanthomonas oryzae
The rice leaf blight bacterium PXO99 A Respectively culturing in single colony NB culture medium of rice streak disease germ RS105 and Bacillus belgii SF327 at 28 deg.C and 180rpm for 12 hr, and uniformly adjusting the concentration of the culture solution to OD 600 2.0. Respectively suck PXO99 A Mixing with 200 μ L of RS105 bacterial liquid and about 25mL of NA solid culture medium, turning the plate, placing Oxford cups with diameter of 6mm at the center of the NA plate, and inscribing 10 μ L of SF327 bacterial liquid in each Oxford cupAnd 3 times of the pathogenic bacteria are repeated, the pathogenic bacteria are placed in an incubator at 28 ℃, the existence of the inhibition zone is observed after 48 hours of culture, the size of the inhibition zone is recorded, and the images are arranged and photographed.
Bacillus belgii SF327 pairs against rice bacterial blight PXO99 A The antagonistic effect against rice streak disease germ RS105 is shown in FIG. 6, the antagonistic effect against rice streak disease germ RS105 is shown in FIG. 7, and the bacteriostatic effect is shown in Table 4.
The results show that Bacillus belgii SF327 pairs Padina bigelovii PXO99 A The bacteriostasis rate of the strain is 75.92 percent, and the bacteriostasis rate to rice streak disease RS105 is 73.93 percent. This indicates that SF327 has a significant antagonistic effect on Xanthomonas oryzae.
TABLE 4 bacteriostatic effect of Bacillus beilesiensis SF327 on Xanthomonas oryzae
2) Determination of antagonistic Activity of Bacillus belgii SF327 against 5 plant pathogenic fungi
Adopting a filter paper sheet opposing culture method. Magnaporthe grisea, Sclerotinia cucumerinum, Phytophthora capsici, colletotrichum gloeosporioides and colletotrichum gloeosporioides are respectively cultured on a PDA culture medium, after hyphae grow over the whole plate, a fungus block is punched on the plate by a hole puncher with a certain aperture, one fungus cake is taken, the hyphae face of the fungus cake faces downwards and is inoculated to the center of a new PDA plate, filter paper sheets are placed on the left and right sides of the fungus cake at equal distance (35mm), the test Bacillus belgii SF 3275 mu L is dripped on the filter paper sheet, each treatment is repeated for 3 times, and meanwhile, the plate which is not connected with the Bacillus belgii SF327 is used as a comparison. The culture was carried out in an incubator at 28 ℃ and after 5 days the bacteriostatic phenomena were observed and recorded.
TABLE 5 bacteriostatic effect of Bacillus beilesiensis SF327 on 5 plant pathogenic fungi
The antagonistic effect of Bacillus belgii SF327 against Pyricularia oryzae is shown in FIG. 8, the antagonistic effect against Cucumis sativus wilt is shown in FIG. 9, the antagonistic effect against Phytophthora capsici is shown in FIG. 10, the antagonistic effect against colletotrichum gloeosporioides is shown in FIG. 11, and the antagonistic effect against colletotrichum gloeosporioides is shown in FIG. 12.
The results show that the bacteriostasis rates of the Bacillus belgii SF327 on rice blast bacteria are 60.91%, on cucumber fusarium wilt bacteria are 64.68%, on phytophthora capsici is 58.57%, and on rubber gum and colletotrichum oxysporum are 59.72% and 67.42%, respectively. This indicates that SF327 has a better antagonistic effect against all 5 pathogenic fungi.
Therefore, the Bacillus belgii SF327 provided by the invention can generate auxin IAA, has a growth promoting effect on Brassica chinensis, has a remarkable antagonistic effect on rice bacterial blight and rice streak germ, has an antagonistic effect on various plant pathogenic fungi which are important in production, and provides new resources for biological control of rice bacterial diseases, fungal diseases and rubber tree anthracnose.
The embodiments described above are described to facilitate an understanding and use of the invention by those skilled in the art. It will be readily apparent to those skilled in the art that various modifications to these embodiments may be made, and the generic principles described herein may be applied to other embodiments without the use of the inventive faculty. Therefore, the present invention is not limited to the above embodiments, and those skilled in the art should make improvements and modifications within the scope of the present invention based on the disclosure of the present invention.
Sequence listing
<110> Shanghai university of transportation
<120> Bacillus belgii SF327 strain and application thereof
<160> 1
<170> SIPOSequenceListing 1.0
<210> 1
<211> 1423
<212> DNA
<213> Bacillus belgii (Bacillus velezensis)
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tgcaagtcga gcggacagat gggagcttgc tccctgatgt tagcggcgga cgggtgagta 60
acacgtgggt aacctgcctg taagactggg ataactccgg gaaaccgggg ctaataccgg 120
atggttgtct gaaccgcatg gttcagacat aaaaggtggc ttcggctacc acttacagat 180
ggacccgcgg cgcattagct agttggtgag gtaacggctc accaaggcga cgatgcgtag 240
ccgacctgag agggtgatcg gccacactgg gactgagaca cggcccagac tcctacggga 300
ggcagcagta gggaatcttc cgcaatggac gaaagtctga cggagcaacg ccgcgtgagt 360
gatgaaggtt ttcggatcgt aaagctctgt tgttagggaa gaacaagtgc cgttcaaata 420
gggcggcacc ttgacggtac ctaaccagaa agccacggct aactacgtgc cagcagccgc 480
ggtaatacgt aggtggcaag cgttgtccgg aattattggg cgtaaagggc tcgcaggcgg 540
tttcttaagt ctgatgtgaa agcccccggc tcaaccgggg agggtcattg gaaactgggg 600
aacttgagtg cagaagagga gagtggaatt ccacgtgtag cggtgaaatg cgtagagatg 660
tggaggaaca ccagtggcga aggcgactct ctggtctgta actgacgctg aggagcgaaa 720
gcgtggggag cgaacaggat tagataccct ggtagtccac gccgtaaacg atgagtgcta 780
agtgttaggg ggtttccgcc ccttagtgct gcagctaacg cattaagcac tccgcctggg 840
gagtacggtc gcaagactga aactcaaagg aattgacggg ggcccgcaca agcggtggag 900
catgtggttt aattcgaagc aacgcgaaga accttaccag gtcttgacat cctctgacaa 960
tcctagagat aggacgtccc cttcgggggc agagtgacag gtggtgcatg gttgtcgtca 1020
gctcgtgtcg tgagatgttg ggttaagtcc cgcaacgagc gcaacccttg atcttagttg 1080
ccagcattca gttgggcact ctaaggtgac tgccggtgac aaaccggagg aaggtgggga 1140
ccagcattca gttgggcact ctaaggtgac tgccggtgac aaaccggagg aaggtgggga 1200
caaagggcag cgaaaccgcg aggttaagcc aatcccacaa atctgttctc agttcggatc 1260
gcagtctgca actcgactgc gtgaagctgg aatcgctagt aatcgcggat cagcatgccg 1320
cggtgaatac gttcccgggc cttgtacaca ccgcccgtca caccacgaga gtttgtaaca 1380
cccgaagtcg gtgaggtaac cttttaggag ccagccgccg aag 1423
Claims (10)
1. The Bacillus belgii is characterized by being named as Bacillus belgii (Bacillus velezensis) SF327, and being preserved in China center for type culture Collection (CCTCC M2022130) at 18 months 2 in 2022.
2. The Bacillus belgii 16SrRNA gene of claim 1, wherein the Bacillus belgii 16S rRNA gene sequence is shown in SEQ ID No. 1.
3. The use of a strain of bacillus beiLeisi as claimed in claim 1, wherein the bacillus beiLeisi has growth promoting effect on brassica oleracea.
4. The use of a strain of Bacillus belgii according to claim 3, wherein the Brassica oleracea is Chondrus macrolepis.
5. The use of a strain of Bacillus belgii according to claim 3, wherein the growth promoting effect comprises the ability to produce indole-3-acetate auxins and to increase the fresh weight of aerial parts of Brassica oleracea.
6. The use of a strain of bacillus beiLensis according to claim 1 as an antagonist of Xanthomonas oryzae (Xanthomonas oryzae).
7. The use of a strain of Bacillus beijerinckii as claimed in claim 6, wherein the Xanthomonas oryzae comprises Xanthomonas oryzae pv. oryzae PXO99 A And rice leaf streak germ (Xanthomonas oryzae pv. oryzae) RS 105.
8. Use of a strain of bacillus beijerinckii according to claim 1 as an antagonistic bacterium against phytopathogenic fungi.
9. The use of a strain of Bacillus belgii as claimed in claim 8, wherein the pathogenic fungi include Magnaporthe oryzae (Magnaporthe oryzae), Fusarium oxysporum f.sp.
10. The use of a strain of bacillus beijerinckii according to claim 1 as a brassica oleracea growth promoting agent, as an antagonist of Xanthomonas oryzae (Xanthomonas oryzae), and as an antagonist of phytopathogenic fungi.
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| CN116904339A (en) * | 2023-04-13 | 2023-10-20 | 山西农业大学 | Preparation and application methods of Bacillus belicus LY7 strain suspending agent |
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| CN116904339A (en) * | 2023-04-13 | 2023-10-20 | 山西农业大学 | Preparation and application methods of Bacillus belicus LY7 strain suspending agent |
| CN118325792A (en) * | 2024-06-12 | 2024-07-12 | 中国热带农业科学院三亚研究院 | Endophytic bacillus bailii LSR7 of rubber tree and application thereof |
| CN118325792B (en) * | 2024-06-12 | 2024-08-09 | 中国热带农业科学院三亚研究院 | Endophytic bacillus bailii LSR7 of rubber tree and application thereof |
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