CN114736821A

CN114736821A - Bacillus belgii SF305 with antagonistic effect on rubber tree red root pathogen and application thereof

Info

Publication number: CN114736821A
Application number: CN202210357384.7A
Authority: CN
Inventors: 涂敏; 邹丽芳; 方园; 伏雪; 阎依超; 蔡海滨
Original assignee: Shanghai Jiaotong University; Rubber Research Institute Chinese Academy Tropical Agricultural Sciences
Current assignee: Shanghai Jiaotong University; Rubber Research Institute Chinese Academy Tropical Agricultural Sciences
Priority date: 2022-04-03
Filing date: 2022-04-03
Publication date: 2022-07-12
Anticipated expiration: 2042-04-03
Also published as: CN114736821B

Abstract

The invention relates to a bacillus beilesensis SF305 strain and application thereof. The preservation number of the Bacillus belgii SF305 is CCTCC NO: m2022129 has a remarkable inhibiting effect on rubber tree red root pathogens, provides a good biocontrol resource for the prevention and treatment of rubber tree red root diseases, and has good application potential in the prevention and treatment of rubber tree red root diseases. Meanwhile, the Bacillus belgii SF305 disclosed by the invention has good antibacterial activity on plant pathogenic bacteria such as colletotrichum gloeosporioides, rice blast, fusarium oxysporum, phytophthora capsici, botrytis cinerea, rice leaf blight, rice streak disease and the like, can promote the growth of crops and increase the fresh weight of the crops, shows that the strain has application prospects of disease prevention and growth promotion, and lays a foundation for research and development of novel microbial agents for diseases such as rubber anthracnose, rice leaf blight, streak disease, rice blast and the like.

Description

Bacillus belgii SF305 with antagonistic effect on rubber tree red root pathogen and application thereof

Technical Field

The invention belongs to the field of application of microbial technology, and particularly relates to bacillus beiLeisi SF305 and application thereof.

Background

Because of its good elasticity, insulating property, strong extensibility, good water-proofing property, air-tightness and wear-resisting property, it is easy to adhere with other materials, and can be extensively used in the fields of military industry, spaceflight, medical treatment and automobile manufacture, etc., it is an important industrial raw material and strategic material, and its product can be up to 7 million (Liu Ruijin, Mo, Yanlin, etc. the recognization and development of strategic position of natural rubber industry in China is suggested, Chinese tropical agriculture 2022(01): 13-18). Natural rubber is used as a key agricultural Raw material in the United states, the European Union lists the natural rubber into a key Raw material list, and China also actively develops the rubber industry after the country is built (European Commission. study on the EU's list of Critical Raw Materials-Final Report (2020). Brussels: European Commission, 2020); more than 98% of the world's natural rubber is produced in Hevea brasiliensis (Hevea brasiliensis) (Chengshui, Mao Xincui. world's natural rubber industry development analysis report. Chinese tropical agriculture. 2020(01):29-34+ 59).

The root infectious disease with the widest harm area and the largest influence on rubber tree production in China is the red root disease, and the pathogenic bacteria of the root infectious disease are Ganoderma pseudoperreum (Jingjing Yu, species identification and biological characteristic research of pathogenic Ganoderma of rubber trees and acacia trees, Hainan university, 2018, Zhangjin, Zhang Xin, and the like. biological culture characteristics of pathogenic bacteria of rubber tree red root disease. report of tropical crops, 2008,29(5):632 + one). The disease damages the root or the root neck of the Brazilian rubber tree, leaves of the rubber tree become small after the rubber tree is infected with the disease, the rubber tree lacks luster, turns yellow and curls, the crown of the tree gradually becomes sparse, the trunk shrinks or the base of the trunk has a ditch, and finally dies, and the production life and the rubber yield of the rubber forest are seriously shortened (Chua Shiyiying, Li Guo. The host range of rubber tree red root disease pathogenic bacteria is wide, and can infect more than ten crops of orange, lichee, tea tree, etc., at the same time, the latent period of root disease pathogenic bacteria in soil is very long (Oekang, Huangzongdao. tropical northern rubber tree cultivation. Guangdong science and technology publishing company, Guangzhou, 1987), the pathogenic bacteria whose root diameter is 3cm can survive for 2 years in soil, and the annual average spreading rate can be up to 1.83 m after meeting suitable host (Zhangqiang, Zhang Qiang. spreading speed of rubber tree red root disease and prediction report. tropical crop academic report, 1998,19(1): 7-12). At present, the red root disease commonly occurs in rubber planting areas in China, the disease areas are almost distributed in each rubber planting farm, the morbidity of forest sections with serious morbidity reaches 40 percent, the morbidity of young tree areas which are not subjected to prevention treatment can reach 60 percent, and if the mortality is not treated in time and reaches 100 percent, huge loss is brought to the rubber industry (hypersusceptibility, Chua seashore, Pentagon, and the like).

For years, the prevention and treatment of rubber tree red root disease in production mainly comprises mechanical excavation and chemical prevention and treatment. Since the 70 th generation of 19 th century, 75% tridemorph emulsifiable concentrate and 15% triazolone wettable powder as main chemical agents were mainly used in the field (forest wu, wu xiang, guozhou, etc., tridemorph for controlling rubber red root disease. pesticide, 1992,31(3): 57-58; Tan Xiang, Fan Hu Xiong. rubber tree red root disease development and control strategy in rubber gardens. plant protection.1991, (06): 46-48). In 2012, the field test on the propiconazole missible oil shows that the control effect of the propiconazole missible oil is equivalent to that of tridemorph, and the propiconazole missible oil is moderate in price. Indoor toxicity determination triazole pesticide hexaconazole WP has the strongest bacteriostasis effect on rubber tree red root pathogen (Liquan, Qijun. propiconazole control rubber tree red root disease field test. tropical biology report 2012.3(4):325 + 328). However, chemical control still has great limitations because of the problems of pesticide residue, environmental pollution, pathogen resistance, reduced control effect and the like.

In recent years, biological control has become one of the hot spots in the field of phytopathology research, and on one hand, the biological control has mild influence on the environment, and on the other hand, the biological control has certain specificity on pathogenic bacteria and is not easy to cause resistance to the pathogenic bacteria. Biocontrol fungus trichoderma has an antibacterial effect on the red-rooted germs (Qinyun, alexandrine, Yangmeifang, and the like. 8 strains of trichoderma have an antagonistic effect on the red-rooted germs of rubber trees. tropical agriculture, 2015, 35(03):41-45+ 53); researches show that the bacillus subtilis CZk1 also has a good inhibition effect on erythrorhizosis (Liyangqiong, Lirui, Wuwei Huai, and the like. the mixed active components of the bacillus subtilis CZk1 volatile matters have an inhibition mechanism on the ganoderma rubberberidis, China agricultural science and technology guide, 2022,24(2): 152-. The red root germ contains polysaccharide and polyphenol, has slow growth speed and is relatively difficult to research the functions of the gene. Therefore, for researchers, the method responds to the national plant protection policy of 'prevention-oriented and comprehensive control', screens out biological resources with antagonistic action on diseases from natural environment, is a control method which is worthy of exploration, and is also a work with great significance.

Disclosure of Invention

The invention aims to provide a bacillus belvesii with a remarkable inhibiting effect on rubber tree red root pathogens and application thereof.

The Bacillus beleisi shows broad-spectrum antibacterial activity on plant pathogenic fungi such as colletotrichum gloeosporioides, colletotrichum oxysporum, rice blast, fusarium oxysporum, phytophthora capsici, botrytis cinerea and the like; has good bacteriostatic activity on bacterial diseases such as rice bacterial blight and rice stripe spot, and can promote plant growth. The result of the invention shows that the strain has application prospects of disease prevention and growth promotion, provides good biocontrol resources for the prevention and control of rubber tree red root disease, and also provides reference for the prevention and control of rubber anthracnose, rice bacterial leaf blight, stripe disease and other diseases.

The technical scheme of the invention is realized as follows:

the first aspect of the invention provides a Bacillus beleisi, which is separated from soil in garden at estuary of Hainan City, Hainan, in 2018, 11 and 8 days, and is named as Bacillus beleisi (Bacillus velezensis) SF305, and the Bacillus beleisi has been preserved in China Center for Type Culture Collection (CCTCC) in 2022, 02 and 18 days, with the preservation number of CCTCC NO: m2022129.

In a second aspect of the present invention, there is provided a fermentation broth of Bacillus belgii as described above.

The preparation method of the fermentation liquor comprises the following steps:

and inoculating the single colony of the Bacillus belgii into an NB culture medium, and culturing in a shaker at 28 ℃ and 180rpm for 12h to obtain the Bacillus belgii.

In a third aspect, the invention provides the application of the bacillus belgii or the fermentation liquid in antagonizing rubber tree red root pathogen.

The fourth aspect of the invention provides the application of the bacillus belgii or the fermentation liquor in preparing a medicament for preventing and treating diseases caused by rubber tree red root pathogen (Ganoderma pseudoterreum).

In a fifth aspect of the present invention, there is provided Bacillus belgii or the fermentation broth having antagonistic action against various phytopathogenic fungi, specifically including Colletotrichum gloeosporioides, Colletotrichum, Magnaporthe oryzae (Magnaporthe oryzae), Fusarium oxysporum f.sp.culumerinum, Phytophthora capsici (Phytophthora capsici) and Botrytis cinerea.

In a sixth aspect, the present invention provides the use of the above-mentioned bacillus belgii or the above-mentioned fermentation broth for antagonizing Xanthomonas oryzae pv. oryzae, Xoo and Xanthomonas oryzae pv. oryzae, Xoc.

In one embodiment of the present invention, the rice bacterial blight comprises PXO99^AThe rice streak disease germ comprises RS 105.

The seventh aspect of the present invention provides the use of the above-mentioned Bacillus belgii or the above-mentioned fermentation broth for the preparation of a preparation for controlling diseases caused by colletotrichum gloeosporioides, Pyricularia oryzae, Cucumis sativus wilt, Phytophthora capsici, Botrytis cinerea, Fusarium oxysporum and/or Rhizoctonia solani.

An eighth aspect of the present invention provides the use of the above-mentioned bacillus belgii or the above-mentioned fermentation broth for promoting plant growth.

The ninth aspect of the invention provides a biocontrol microbial inoculum, which comprises the bacillus belgii or the fermentation liquor.

The invention has the beneficial effects that:

compared with the prior art, the Bacillus belgii SF305 and the fermentation liquor thereof have obvious antagonistic action on rubber tree red root pathogenic bacteria and antagonistic activity on plant pathogenic fungi and bacteria which are important in production at present. The result of the invention shows that the strain has broad-spectrum antibacterial and growth promoting capability, has better application potential in preventing and treating rubber tree root diseases and various important fungal and bacterial diseases of crops, has biological control potential to be further developed, and lays a foundation for developing multifunctional biological control agents.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.

FIG. 1 is a microscope (1000X) photograph, an electron micrograph and colony morphology of Bacillus beleisi SF 305; wherein, a is a microscope (1000X) observation picture, b is a colony morphology plate picture, and c is a scanning electron microscope observation picture.

FIG. 2.16 gel electrophoresis results of S rRNA gene. Wherein 1 represents DL2000 Marker; 2 represents the product of the 16S rRNA gene.

FIG. 3 shows the result of alignment of the gene sequences of Bacillus belgii SF 30516S rRNA, which is a phylogenetic tree of Neighbor-Joining constructed using Virgibacillus pantothecus NCDO 1765(X60627) as an outgrowth.

FIG. 4 is a graph showing the antagonistic effect of Bacillus belgii SF305 against Ralstonia koenigii.

FIG. 5 is a graph showing the antagonistic effect of Bacillus belgii SF305 against colletotrichum lecanii.

FIG. 6 is a graph showing the antagonistic effect of Bacillus belgii SF305 against colletotrichum gloeosporioides.

FIG. 7 is a graph showing the antagonistic effect of Bacillus belgii SF305 against Pyricularia oryzae.

FIG. 8 is a graph showing the antagonistic effect of Bacillus belgii SF305 against cucumber fusarium oxysporum.

FIG. 9 is a graph showing the antagonistic effect of Bacillus belgii SF305 against Phytophthora capsici.

FIG. 10 is a graph showing the antagonistic effect of Bacillus belgii SF305 on Botrytis cinerea.

FIG. 11 is a graph showing the antagonistic effect of Bacillus belgii SF305 on rice bacterial blight strain PXO 99A.

FIG. 12 is a graph showing the antagonistic effect of Bacillus belgii SF305 on rice streak disease germ RS 105.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present 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, adding water to dissolve the mixture, fixing the volume to 1000mL, adjusting the pH value to 7.0-7.2, and sterilizing under high pressure (121 ℃,20 min).

NB liquid medium (g/L): 3g of beef extract, 5g of polypeptone, 10g of sucrose and 1g of yeast powder, dissolving in water, diluting to 1000mL, adjusting the pH value to 7.0-7.2, and autoclaving (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).

Example 1 preparation of Bacillus belgii SF305

1. Source of soil

Soil of the garden at the Hongkong estuary of Haikou city of Hainan province collected 11, 8 and 2018.

2. Screening of strains

(1) Soil sample collection

Collecting soil samples of about 10cm of the surface layer of the root circumference 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 an erlenmeyer flask, adding 90mL of sterile water, oscillating in a shaking table at 200 rpm 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 each⁰、10^-1、10^-2、10^-3、10^-4、10^-5Total 6 dilutions. 200. mu.L of each bacterial suspension dilution was uniformly spread on NA plates containing rice leaf streak disease RS105(OD600 ═ 2.0) and each gradient was repeated 3 times. Placing the plate at 28 deg.C for cultureCulturing in a box for 48h, and observing.

(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 fermentation liquor and 1mL of 50% sterile glycerol, slightly shaking and uniformly mixing, and storing at-80 ℃ for a long time.

Finally obtaining Bacillus velezensis (Bacillus velezensis) SF305, which is preserved in China Center for Type Culture Collection (CCTCC) at 18.02/2022 with the preservation number of CCTCC NO: m2022129, address Wuhan university in Wuhan, China.

The observation photograph, the electron micrograph and the colony morphology of the obtained Bacillus belgii SF305 under a microscope (1000X) are shown in FIG. 1.

Example 2 16S rRNA of Bacillus belgii SF305

The genomic DNA of the strain SF305 was extracted using the bacterial DNA Kit of Omega, and the genomic DNA of the strain was purified using bacterial 16S rDNA gene primers: 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:

TABLE 1 Ex-Taq polymerase chain reaction System

The basic conditions of the PCR reaction are as follows: pre-denaturation at 95 deg.C for 5min, denaturation at 94 deg.C for 1min, annealing at 56 deg.C for 1min, extension at 72 deg.C for 2min (1kb/min), pre-extension at 72 deg.C for 6min, and storage at 4 deg.C for 30 cycles. After the reaction, the PCR products were checked by 1% agarose gel electrophoresis, and the PCR stock solutions were sent to Shisanha Biotechnology (Shanghai) Co., Ltd for sequencing. The sequencing results are shown in FIG. 2. And (3) analyzing and sequencing results by using the DNA Star, and performing BLAST comparison on an NCBI website to determine the species of the related strains of bacteria. As shown in table 2. The sequence of the 16SrRNA gene of Bacillus belgii SF305 is shown in SEQ ID NO. 1.

TABLE 2 B. B.baileyi SF 30516S rRNA Gene sequencing BLAST results

The sequence alignment result of Bacillus belgii SF 30516S rRNA gene is shown in FIG. 3, and the result shows that the Neighbor-Joining phylogenetic tree constructed by using Virgibacillus pantothenticatus NCDO 1765(X60627) as an outer branch: the 16S rRNA gene of SF305 has 99.36% similarity to Bacillus velezensis.

Example 3 physiological and biochemical identification of Bacillus beilesiensis SF305

The physiological and biochemical characteristics of the bacillus beleisi SF305 are as follows: 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; unable to oxidize glucose, mannitol, inositol, sorbitol, rhamnose, etc.; can produce acid by using 20 carbon sources such as glycerol, L-arabinose, ribose, D-xylose, galactose, glucose and fructose, and the like, and is shown in tables 3 and 4.

TABLE 3 physiological and biochemical characteristics of Strain SF 305-enzyme Activity, carbon Source Oxidation

+: positive reaction; -: negative reaction; w: weak positive reaction

TABLE 4 physiological and biochemical characteristics of Strain SF305 acid production Using carbon sources

+: positive reaction; -: negative reaction; w: weak positive reaction

Example 4 antagonistic Spectrometry of Bacillus beilesiensis SF305

Inoculating the single colony of the bacillus belgii SF305 into an NB culture medium, and culturing for 12h in a shaker at 28 ℃ and 180rpm to obtain the fermentation liquor of the bacillus belgii SF 305. Then, the fermentation broth was adjusted to an OD600 of 2.0 for future use.

1) Determination of antagonistic Activity of Bacillus belgii SF305 against Ralstonia koenigii

Taking rubber tree red root germ (Ganoderma pseudorhizosphere) fungus cakes on a PDA culture medium, after hyphae grow over the whole flat plate, punching fungus blocks on the flat plate by using a puncher with the aperture of 5mm, taking one fungus cake, inoculating the fungus cake to the center of a new PDA flat plate, and putting pretreated filter paper sheets on the upper side, the lower side, the left side and the right side of the fungus cake at equal distance (20 mm). Wherein the filter paper sheets at the upper and lower positions are sterilized, dried and added with 30ul of NB culture medium, the filter paper sheets at the left and right positions are sterilized, dried and added with 30ul of Bacillus belief SF305 fermentation liquor, and 3 times of repetition are set. Meanwhile, a plate which is not inoculated with Bacillus belgii SF305 fermentation liquor and is only inoculated with rubber tree red root germ cakes is taken as a reference. And (5) placing the mixture in an incubator at 28 ℃, and observing and recording the bacteriostasis result after culturing for 7 d.

FIG. 4 shows antagonistic effect of Bacillus belgii SF305 against rubber tree red root pathogen, and Table 5 shows bacteriostatic effect.

The result shows that the average bacteriostasis rate of the Bacillus belgii SF305 to the rubber tree red root pathogen is 73.76%, which indicates that the SF305 has obvious antagonism to the rubber tree red root pathogen.

TABLE 5 bacteriostatic effect of Bacillus belgii SF305 on rubber tree red root pathogen

2) Determination of antagonistic Activity of Bacillus beilesiensis SF305 against 6 plant pathogenic fungi

Respectively culturing colletotrichum gloeosporioides, rice blast, cucumber wilt, phytophthora capsici and botrytis cinerea on a PDA culture medium by adopting a filter paper sheet opposing culture method, punching fungus blocks on a flat plate by using a puncher with the aperture of 5mm after hypha grows over the whole flat plate, taking one of the fungus cakes, inoculating the fungus cake to the center of a new PDA flat plate with the hypha facing upwards, and putting pre-treated filter paper sheets on the fungus cake at equal intervals (20mm) of the upper part, the lower part, the left part and the right part. Wherein the filter paper sheets at the upper and lower positions are sterilized, dried and added with 30ul of NB culture medium, the filter paper sheets at the left and right positions are added with 30ul of Bacillus belief SF305 fermentation liquor, and each pathogenic bacterium is repeated for 3 times. Meanwhile, plates which are not connected with Bacillus belgii SF305 fermentation liquor but only connected with 6 pathogenic fungus cakes of colletotrichum gloeosporioides, rice blast, fusarium oxysporum, phytophthora capsici and botrytis cinerea are used as controls. After 5 days of culture at 25 ℃, the bacteriostatic phenomena were observed and recorded. The results are shown in Table 6 and FIGS. 5 to 10.

TABLE 6 bacteriostatic effect of Bacillus belgii SF305 on 6 plant pathogenic fungi

The antagonistic effect of Bacillus belgii SF305 on colletotrichum gloeosporioides is shown in FIG. 5, on colletotrichum gloeosporioides is shown in FIG. 6, on Magnaporthe grisea is shown in FIG. 7, on Fusarium oxysporum is shown in FIG. 8, on Phytophthora capsici is shown in FIG. 9, and on Botrytis cinerea is shown in FIG. 10.

The results show that the bacteriostasis rate of the Bacillus belgii SF305 to rubber tree spore colletotrichum is 58.70%, the bacteriostasis rate to rubber tree tip spore colletotrichum is 65.36%, the bacteriostasis rate to rice blast is 62.04%, the bacteriostasis rate to cucumber fusarium wilt is 60.55%, the bacteriostasis rate to phytophthora capsici is 61.85%, and the bacteriostasis rate to botrytis is 57.41%. This indicates that SF305 has a good antagonistic effect against all 6 pathogenic fungi.

3) Determination of antagonistic activity of Bacillus beilesiensis SF305 against Xanthomonas oryzae

The bacterial blight of rice (Xanthomos oryzae pv. oryzae, Xoo) PXO99^AAnd rice leaf blight bacterium (Xanthomonas oryzae pv. oryzae Zicola, Xoc) RS105 in single colony NB medium, respectively, shaking-culturing at 28 deg.C and 180rpm for 12h to obtain rice leaf blight bacterium PXO99^AAnd fermentation liquor of rice streak disease germ RS 105. Then the concentration of the fermentation liquor is uniformly adjusted to OD₆₀₀＝2.0。

Respectively suck PXO99^AAnd mixing 200 μ L of RS105 fermentation liquor with about 25mL of NA solid culture medium, inverting, placing oxford cups with the diameter of 6mm in the center of the NA plate, and inscribing 10 μ L of SF305 fermentation liquor in each oxford cup, wherein each pathogenic bacterium is repeated for 3 times. Meanwhile, the fermentation liquor of Bacillus beiLeisi SF305 is not inoculated, and only PXO99 is inoculated^AAnd plates of RS105 fermentation broth as controls. And (3) placing the mixture in an incubator at 28 ℃, observing whether the inhibition zone exists or not after culturing for 48 hours, recording the size of the inhibition zone, and finishing and photographing.

Bacillus belgii SF305 for Paddy rice bacterial blight PXO99^AThe antagonistic effect of (b) is shown in fig. 11, the antagonistic effect against rice streak blotch virus RS105 is shown in fig. 12, and the bacteriostatic effect is shown in table 7.

The results show that Bacillus beilesiensis SF305 is used for rice bacterial blight strain PXO99^AThe bacteriostasis rate of the strain is 68.82 percent, and the bacteriostasis rate to rice streak disease RS105 is 68.51 percent. This indicates that SF305 has a good antagonistic effect on Xanthomonas oryzae.

TABLE 7 bacteriostatic effect of Bacillus beilesiensis SF305 on Xanthomonas oryzae

Therefore, the Bacillus belgii SF305 provided by the invention has a remarkable antagonistic effect on rubber tree red root pathogens, has an antagonistic effect on various production important plant pathogenic fungi and bacteria, provides a new resource for biological control of rubber tree red root pathogens, and lays a foundation for research and development of novel microbial agents for other important disease control.

Example 5 growth-promoting assay for Bacillus belgii SF305

Bacillus belgii SF305 was individually cultured in NB medium as single colony at 28 ℃ for 12 hours in a shaker at 180rpm, and then diluted 5-fold after adjusting the fermentation broth OD600 to 1.0.

Selecting small green vegetable 'No. two long' vegetable seedlings which are sown at the same time and have similar growth vigor, soaking the vegetable seedlings in diluted Bacillus beilaisi SF305 fermentation liquor for 5min, transplanting, and directly transplanting the vegetable seedlings without any treatment in a control group. And uniformly performing field cultivation management. After 25 days of cultivation, the overground parts of the small green vegetables are picked respectively (the underground parts of the small green vegetables are removed by scissors), and the small green vegetables are weighed every 4 vegetables immediately, and the fresh weight increasing rate of the overground parts are calculated.

TABLE 8 growth promoting and weight increasing effects

As can be seen from Table 8, the fresh weight of the brassica chinensis can be significantly increased by using the Bacillus baileyi SF305 strain fermentation broth for root soaking treatment before transplanting. Thus, the Bacillus belgii SF305 has a remarkable growth promoting effect.

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> rubber institute of tropical agricultural academy of sciences of China

Shanghai Jiaotong University

<120> Bacillus belgii SF305 having antagonism to rubber tree red root germ and application thereof

<160> 1

<170> SIPOSequenceListing 1.0

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<211> 1450

<212> DNA

<213> Bacillus beilaissensis SF305(Bacillus velezensis SF305)

<400> 1

ggatggcgcg tgctatactg caagtcgagc ggacagatgg gagcttgctc cctgatgtta 60

gcggcggacg ggtgagtaac acgtgggtaa cctgcctgta agactgggat aactccggga 120

aaccggggct aataccggat ggttgtttga accgcatggt tcagacataa aaggtggctt 180

cggctaccac ttacagatgg acccgcggcg cattagctag ttggtgaggt aacggctcac 240

caaggcgacg atgcgtagcc gacctgagag ggtgatcggc cacactggga ctgagacacg 300

gcccagactc ctacgggagg cagcagtagg gaatcttccg caatggacga aagtctgacg 360

gagcaacgcc gcgtgagtga tgaaggtttt cggatcgtaa agctctgttg ttagggaaga 420

acaagtgccg ttcaaatagg gcggcacctt gacggtacct aaccagaaag ccacggctaa 480

ctacgtgcca gcagccgcgg taatacgtag gtggcaagcg ttgtccggaa ttattgggcg 540

taaagggctc gcaggcggtt tcttaagtct gatgtgaaag cccccggctc aaccggggag 600

ggtcattgga aactggggaa cttgagtgca gaagaggaga gtggaattcc acgtgtagcg 660

gtgaaatgcg tagagatgtg gaggaacacc agtggcgaag gcgactctct ggtctgtaac 720

tgacgctgag gagcgaaagc gtggggagcg aacaggatta gataccctgg tagtccacgc 780

cgtaaacgat gagtgctaag tgttaggggg tttccgcccc ttagtgctgc agctaacgca 840

ttaagcactc cgcctgggga gtacggtcgc aagactgaaa ctcaaaggaa ttgacggggg 900

cccgcacaag cggtggagca tgtggtttaa ttcgaagcaa cgcgaagaac cttaccaggt 960

cttgacatcc tctgacaatc ctagagatag gacgtcccct tcgggggcag agtgacaggt 1020

ggtgcatggt tgtcgtcagc tcgtgtcgtg agatgttggg ttaagtcccg caacgagcgc 1080

aacccttgat cttagttgcc agcattcagt tgggcactct aaggtgactg ccggtgacaa 1140

accggaggaa ggtggggatg acgtcaaatc atcatgcccc ttatgacctg ggctacacac 1200

gtgctacaat ggacagaaca aagggcagcg aaaccgcgag gttaagccaa tcccacaaat 1260

ctgttctcag ttcggatcgc agtctgcaac tcgactgcgt gaagctggaa tcgctagtaa 1320

tcgcggatca gcatgccgcg gtgaatacgt tcccgggcct tgtacacacc gcccgtcaca 1380

ccacgagagt ttgtaacacc cgaagtcggt gaggtaacct ttatggagcc agccgccgaa 1440

gggaccagag 1450

Claims

1. The Bacillus belgii is Bacillus belgii (Bacillus velezensis) SF305, is preserved in China Center for Type Culture Collection (CCTCC) at 18/02/2022, and has a preservation number of CCTCC NO: m2022129.

2. A fermentation broth of bacillus belgii of claim 1.

3. The method of claim 2, comprising the steps of:

the Bacillus belgii of claim 1, wherein a single colony of the Bacillus belgii is inoculated into NB medium and cultured in a shaker at 28 ℃ and 180rpm for 12 hours.

4. Use of a bacillus beiLeisi as claimed in claim 1 and/or a fermentation broth as claimed in claim 2 for antagonising the species Rhodosporidium hevea (Garoderma pseudoterreum).

5. Use of a bacillus beijerinckii according to claim 1 and/or a fermentation broth according to claim 2 for the preparation of a medicament for controlling diseases caused by rubber tree red root pathogens.

6. Use of a bacillus beijerinckii according to claim 1 and/or a fermentation broth according to claim 2 for antagonising a phytopathogenic fungus comprising: colletotrichum gloeosporioides (Colletotrichum gloeosporioides), Colletotrichum (Colletotrichum aculatum), Magnaporthe oryzae (Magnaporthe oryzae), Fusarium oxysporum (Fusarium oxysporum f.sp. cummerinum), Phytophthora capsici (Phytophthora capsici), and Botrytis cinerea (Botrytis cinerea).

7. Use of a bacillus beijerinckii according to claim 1 and/or a fermentation broth according to claim 2 for antagonism of fusarium oxysporum f.oryzae (Xoo) and alternaria striata (Xanthomonas oryzae pv. oryzae, Xoc); preferably, the rice bacterial leaf blight includes PXO99^AThe rice streak disease germ comprises RS 105.

8. Use of a Bacillus belgii according to claim 1 and/or a fermentation broth according to claim 2 for the preparation of a formulation for the control of diseases caused by colletotrichum hevea, colletotrichum glaucophyllum, pyricularia grisea, fusarium oxysporum, phytophthora capsici, botrytis cinerea, fusarium oxysporum and/or fusarium graminearum.

9. Use of a bacillus beijerinckii according to claim 1 and/or a fermentation broth according to claim 2 for promoting plant growth.

10. A biocontrol microbial agent comprising Bacillus belgii of claim 1 and/or the fermentation broth of claim 2.