CN116254199B - Bacillus bailii JB23 for preventing and controlling sugarcane fungal diseases and application thereof - Google Patents
Bacillus bailii JB23 for preventing and controlling sugarcane fungal diseases and application thereof Download PDFInfo
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Abstract
The invention discloses bacillus bailii JB23 for preventing and controlling sugarcane fungal diseases and application thereof, and belongs to the technical field of crop disease prevention and control. The strain is preserved in China Center for Type Culture Collection (CCTCC) with the preservation time of 2022, 7 months and 18 days and the preservation number of CCTCC NO: m20221134. According to the invention, the bacillus beliensis (Bacillus velezensis) JB23 strain is separated from the sugarcane stem tissue, and the flat plate opposite experiment proves that the bacillus beliensis JB23 strain has a good function of inhibiting the growth of 7 sugarcane pathogenic fungi, and the greenhouse pot experiment proves that the bacillus beliensis JB23 strain has a good application effect on the biological control of sugarcane fungal diseases, can be used for the biological control of sugarcane fungal diseases, and provides a new strain resource for a sugarcane disease biocontrol strain resource library.
Description
Technical Field
The invention relates to the technical field of crop disease control, in particular to bacillus bailii JB23 for controlling sugarcane fungal diseases and application thereof.
Background
Sugarcane is the most important sugar crop in China, and the industrial development of the sugarcane is closely related to the national sugar safety. However, in the production process, the problems of fungus diseases such as sugarcane smut, tip rot and root rot cause the yield and quality of sugarcane to be reduced, and huge economic loss is caused. It is reported that sugarcane smut flows to cause 2-2.5 ten thousand tons of yield loss, and direct economic loss can reach as high as 1025 ten thousand yuan; the sugarcane tip rot causes 15-30 t/hm 2 of yield reduction in a sugarcane area, and the sucrose content is reduced by 0.56%; the most obvious characteristic of the sugarcane smut is that the whole sugarcane cannot normally grow due to the fact that the sugarcane tip head is provided with a black whip in a downward roll, so that the yield of the whole sugarcane is reduced in a large area. The sugarcane tip rot mainly occurs at tender leaf parts at the tips of the sugarcane, infected leaves can be twisted together, when the disease is serious, the growth points at the tips can rot, the tender leaves are necrotic, and even the whole sugarcane is dead. The sugarcane root rot damages the sugarcane root system, so that the growth of the sugarcane root system is blocked and even decayed, the yield is reduced due to the fact that plants cannot absorb nutrients from the root system, the disease latency period is long, and the yield is reduced in a large area once symptoms are displayed. The sustainable development of sugarcane industry has a great risk challenge, and therefore, the prevention and treatment work of sugarcane fungal diseases deserves attention.
The method mainly comprises breeding cultivation disease-resistant varieties, introduction quarantine, seed stem disinfection, chemical agent control, scientific field management, biological control and the like in production for controlling the sugarcane fungus diseases. The disease-resistant variety is most effective at all, but the problem of sudden disease is difficult to solve rapidly for a long breeding time. The pesticide control is quick and efficient, but the irregular application in production causes the problems of enhanced pathogen resistance, safety risk of agricultural products, environmental pollution and the like. In addition, the higher labor cost in production makes scientific field management implementation difficult. Therefore, as a novel control means with environmental protection, safety, reliability, cost saving and synergy, biological control becomes a research hot spot in recent years, and the application prospect is very broad.
Bacillus belicus (Bacillus velezensis) belongs to the genus Bacillus, is gram positive bacteria, is aerobic, has biofilm formation and exercise capacity, and can secrete abundant metabolic substances. Bacillus bailii can colonize plant bodies to generate antibacterial substances and plant biomass, so that the bacillus bailii has great development potential as a biocontrol microbial inoculum for controlling plant diseases. Therefore, the invention screens and obtains a new strain with broad-spectrum antibacterial property, and provides a new idea for biological control of sugarcane mycosis.
Disclosure of Invention
The invention aims to provide bacillus bailii JB23 for preventing and controlling sugarcane fungal diseases and application thereof, so as to solve the problems in the prior art, and the strain inhibits the growth of various sugarcane pathogenic fungi, has good application effect on the biological prevention and control of the sugarcane fungal diseases, can be used for the biological prevention and control of the sugarcane fungal diseases, and provides new strain resources for a sugarcane disease biocontrol strain resource library.
In order to achieve the above object, the present invention provides the following solutions:
The invention provides a bacillus belgium (Bacillus velezensis) JB23 strain which is preserved in China center for type culture collection, wherein the preservation time is 2022, 7 months and 18 days, the preservation address is China center for type culture collection, the university of Wuhan in Wuhan district, wuhan, hubei province, and the preservation number is CCTCC NO: m20221134.
The invention also provides a microbial inoculum comprising the bacillus bailii JB23 strain or the fermentation broth thereof.
Further, the concentration of Bacillus bailii JB23 in the fermentation broth is more than or equal to 10 8 cfu/mL.
The invention also provides application of the bacillus belicus JB23 strain or the microbial inoculum in inhibiting the growth of sugarcane pathogenic fungi.
Further, the sugarcane pathogenic fungi are one or more of sugarcane root rot fungi, sugarcane tip rot fungi and sugarcane smut fungi.
Further, the sugarcane root rot fungi include fusarium shared (Fusarium commune); the fusarium saccarium includes fusarium saccarium (Fusarium sacchari), fusarium solani (Fusarium proliferatum), fusarium neoformans (Fusarium andiyazi), fusarium moniliforme (Fusarium verticillioides) and fusarium oxysporum (Fusarium oxysporum); the sugarcane smut bacteria comprise sugarcane smut bacteria (Sporisorium scitamineum).
The invention also provides application of the bacillus belicus JB23 strain or the microbial inoculum in medicaments for preventing and treating sugarcane root rot, tip rot and smut.
The invention also provides application of the bacillus bailii JB23 strain or the microbial inoculum in preparing medicaments for preventing and treating sugarcane fungal diseases.
Further, the sugarcane fungal diseases include diseases caused by infection of sugarcane roots, stems and leaves by the sugarcane pathogenic fungi.
The invention also provides a method for preventing and controlling the fungal diseases of the sugarcane, and the bacillus beijerinus JB23 strain or the microbial inoculum is applied to the sugarcane.
Further, the sugarcane fungal diseases include sugarcane root rot, tip rot and smut.
The invention discloses the following technical effects:
According to the invention, the bacillus beliensis (Bacillus velezensis) JB23 strain is separated from the sugarcane stem tissue, the flat plate counter test proves that the bacillus beliensis JB23 strain has a good function of inhibiting the growth of 7 sugarcane pathogenic fungi, and the greenhouse pot test proves that the bacillus beliensis JB23 strain has a good application effect on biological control of sugarcane smut, sugarcane tip rot and sugarcane root rot, can be used for biological control of sugarcane fungal diseases, and provides new strain resources for a sugarcane disease biocontrol bacterial resource library.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are needed in the embodiments 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 other drawings may be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a colony and a photomicrograph of Bacillus bailii JB23 strain: wherein, figure a is a front view of the initial colony morphology of the JB23 strain cultured on an LB agar medium at 37 ℃, figure b is a microscopic morphology of the JB23 strain (magnified 100 times by an objective lens), and figure c is a gram staining result of the JB23 strain;
FIG. 2 is a phylogenetic tree of Bacillus bailii JB23 constructed in accordance with the present invention;
FIG. 3 shows the results of the plate-stand cultivation of Bacillus bailii JB23 strain OD 600 at 1 against Fusarium saccarium YN28 (Fusarium neoformans Fusarium andiyazi), fusarium saccarium CNO1 (Fusarium saccarium Fusarium sacchari), fusarium saccarium BS6-2 (Fusarium oxysporum Fusarium oxysporum), fusarium saccarium FN (Fusarium moniliforme Fusarium verticillioides), fusarium saccarium YN41 (Fusarium layering Fusarium proliferatum), fusarium saccarium BS46 (Fusarium shared Fusarium commune), and smut (Fusarium saccarium Sporisorium scitamineum);
FIG. 4 shows the results of the culture of Bacillus belicus JB23 strain OD 600 at a value of 2 against plates of Fusarium oxysporum YN28 (Fusarium neoformans Fusarium andiyazi) and Fusarium oxysporum CNO1 (Fusarium saccarium Fusarium sacchari);
FIG. 5 shows the results of a Bacillus JB23 potting: wherein figure a is a treatment group ①: soaking the sugarcane roots subjected to sterilization water treatment for 30 minutes; figure b is process set ②: sugarcane roots immersed in 2X 10 6 CFU/mL BS46 spore liquid for 30 minutes; figure c is process set ③: 2X 10 6 CFU/mL BS46 spore liquid after soaking for 30 minutes soaks the root of sugarcane with JB23 biocontrol bacteria liquid with OD 600 =1 for 30 minutes.
Detailed Description
Various exemplary embodiments of the invention will now be described in detail, which should not be considered as limiting the invention, but rather as more detailed descriptions of certain aspects, features and embodiments of the invention.
It is to be understood that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. In addition, for numerical ranges in this disclosure, it is understood that each intermediate value between the upper and lower limits of the ranges is also specifically disclosed. Every smaller range between any stated value or stated range, and any other stated value or intermediate value within the stated range, is also encompassed within the invention. The upper and lower limits of these smaller ranges may independently be included or excluded in the range.
Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference for the purpose of disclosing and describing the methods and/or materials associated with the documents. In case of conflict with any incorporated document, the present specification will control.
It will be apparent to those skilled in the art that various modifications and variations can be made in the specific embodiments of the invention described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from consideration of the specification of the present invention. The specification and examples of the present invention are exemplary only.
As used herein, the terms "comprising," "including," "having," "containing," and the like are intended to be inclusive and mean an inclusion, but not limited to.
The invention is further illustrated in the following drawings and specific examples, which are not intended to limit the invention in any way. Unless specifically stated otherwise, the reagents, methods and apparatus employed in the present invention are those conventional in the art.
The sugarcane top rot fungi YN28 (Fusarium neoformans Fusarium andiyazi), sugarcane top rot fungi CNO1 (Fusarium saccarium Fusarium sacchari), sugarcane top rot fungi BS6-2 (Fusarium oxysporum Fusarium oxysporum), sugarcane top rot fungi FN (Fusarium moniliforme Fusarium verticillioides), sugarcane top rot fungi YN41 (Fusarium layering Fusarium proliferatum), sugarcane root rot fungi BS46 (Fusarium sharpleasure Fusarium commune), and sugarcane smut (Saccharum sinensis Roxb. Stigma Sporisorium scitamineum) used in the following examples have been disclosed in the following documents:
1)Muqing Zhang,John Martin Jerome Jeyakumar,2018.Fusarium SpeciesComplex Causing Pokkah Boeng in China:Fusarium-Plant Diseases,Pathogen Diversity,Genetic Diversity,Resistance and Molecular Markers,Chapter 9:139-154.
2)Jihua Wang,Zhe Chai,Bao Yixue,Li Yisha,Haixun Wang,G P RAO,Mu-qing,2018.ZhangFirst Report of Fusarium commune Causing RootRot Disease of Sugarcane(var.Badila)in China.Plant Disease 102(8):
DOI:10.1094/PDIS-07-17-1011-PDN
3)Lu S,Shen X,Chen B.Development of an efficient vector system for geneknock-out and near in-cis gene complementation in the sugarcane smutfungus.Sci Rep.2017Jun 8;7(1):3113.
EXAMPLE 1 isolation of strains
Bacillus belicus JB23 strain is a bacillus belicus obtained by tissue separation of sugarcane stalk part in month 6 of 2020.
The method for separating bacillus bailii JB23 strain comprises the following steps:
(1) Cleaning dust on the surface of sugarcane with purified water, wiping water, and cutting into small blocks with the length of 2cm and the thickness of 2mm in an ultra-clean workbench;
(2) Soaking the cut pieces of sugarcane in absolute ethyl alcohol for 1min, and rinsing with sterilized water for 3 times for 1min each time;
(3) After the water on the sugarcane blocks is absorbed by using sterilizing and mirror wiping paper, the sugarcane blocks are stuck on an LB agar plate for dark culture at 37 ℃ for 48 hours;
(4) And separating, purifying and preserving strains according to the size and the morphology of colonies growing on the culture medium.
Example 2 identification of strains
First, observation of strain morphology was performed. The strain was streaked, cultured at 37℃for 24 hours, and colony characteristics were recorded. And a small amount of thalli is picked and smeared on a glass slide, and is observed under a microscope after being dyed by a gram dyeing method. As a result, in the graph 1 a-c, the JB23 strain is gram positive bacillus, the initial colony form is semitransparent round, the surface is smooth and convex, the colony form is more regular, the later colony form is milky and wrinkled, mucus is secreted, and pigment is not produced in the growth process.
Secondly, the physiological and biochemical characteristics of the strain are tested by referring to the common bacteria System identification Manual. The physiological and biochemical characteristic test results of the bacillus bailii JB23 strain are shown in Table 1. As shown in Table 1, bacillus bailii JB23 strain did not grow at 4℃and was able to grow in the range of 20-42℃and the tolerance concentration to sodium salt was 0-6%. The method can utilize glucose, fructose, sucrose, starch and other carbon sources, has positive effects in catalase reaction and starch hydrolysis test, and can liquefy gelatin.
Table 1 physiological and Biochemical test results of JB23 Strain
Then, through second-generation resequencing and NCBI comparison, the homology between the resequencing sequence of the JB23 strain and Bacillus bailii (Bacillus velezensis) reaches 81.04 percent.
Through the comparison of the evolutionary trees, the relationship between JB23 and other bacillus beleiensis is found: phylogenetic trees were constructed using neighbor-joining (NJ) and Bayesian Inference (BI), see FIG. 2.
Comprehensive morphological and physiological tests, secondary re-sequencing results and evolutionary tree comparison analysis, the results show that the strain belongs to bacillus beleiensis (Pseudomonas mosselii). The bacillus belicus JB23 strain is preserved in China center for type culture collection (China center for type culture collection), wherein the preservation time is 2022, 7 months and 18 days, the preservation address is the university of Wuhan, hubei province, and the preservation number is CCTCC NO: m20221134.
EXAMPLE 3 plate antagonism test of Bacillus bailii JB23 Strain against pathogenic fungi
The antagonism of bacillus beijerinckii JB23 strain to sugarcane pathogenic fungi is measured by a plate confronting method.
The culture medium is PDA culture medium (pH=5.4-5.8), and comprises the following components: potato soaked powder 6g/L, glucose 20g/L, agar 20g/L, and autoclaved at 115 ℃ for 20min.
Preparation of bacillus bailii JB23 bacterial liquid: the biocontrol strain stored at-80 ℃ is inoculated into 1mL of LB broth culture medium according to the proportion of 1:500, and is subjected to shaking culture at 37 ℃ and 220r/min overnight to obtain seed liquid. The seed liquid is dipped by a sterile inoculating loop and streaked on LB agar medium, and is cultured overnight at 37 ℃ to obtain fresh single colony; and (3) picking single bacterial colonies, inoculating the single bacterial colonies into an LB broth culture medium, and culturing for 24 hours at 37 ℃ and 220r/min to obtain fresh biocontrol bacterial liquid.
Antagonism test of JB23 strain and 7 sugarcane pathogenic fungi is carried out by a plate counter culture method. Inoculating activated sugarcane pathogenic fungi cakes (d=6mm) at the central position of the PDA culture medium, taking the central point of the culture medium as an intersection point to make two straight lines which are perpendicular to each other, marking the position 20mm away from the central intersection point on the straight lines, and placing sterile filter paper sheets on the positions; 10 mu L of JB23 strain fermentation liquor is dripped on a sterile filter paper sheet, and the filter paper sheet is placed for 30min and then placed in a 28 ℃ incubator for inversion culture, and each treatment is repeated three times. After 4 days of culture, the diameters of pathogenic bacterial colonies of the treatment group and the control group were measured to determine antagonistic effects of bacillus bailii JB23 strain. The results are shown in FIG. 3, FIG. 4 and Table 2.
As can be seen from fig. 3 and 4, bacillus berryis JB23 strain has remarkable growth inhibitory effects on fusarium saccarium YN28 (fusarium neoformans Fusarium andiyazi), fusarium saccarium CNO1 (fusarium saccarium Fusarium sacchari), fusarium saccarium BS6-2 (fusarium oxysporum Fusarium oxysporum), fusarium saccarium FN (fusarium moniliforme Fusarium verticillioides), fusarium saccarium YN41 (fusarium solani Fusarium proliferatum), fusarium saccarium BS46 (fusarium shared Fusarium commune), and smut saccharum (fusarium saccarium Sporisorium scitamineum).
Meanwhile, the antibacterial rate of the JB23 strain on the pathogenic fungi is measured, and is shown in Table 2. The bacillus belicus JB23 strain has a wide bacteriostasis spectrum, and has good antagonistic capability on various sugarcane pathogenic fungi when the OD 600 value is 1. The colony diameters of the treated groups all showed significant differences at the level of 1% compared to the control group. Then, the OD 600 value of the bacillus bailii JB23 strain is increased to 2, and when the OD 600 value is 1, the fusarium CNO1 and YN28 with the highest inhibition rate are antagonized, whether the inhibition rate is increased or not is observed, and the result shows that the inhibition rate is increased to a certain extent when the OD 600 is 2.
Table 2 antibacterial Rate of JB23 Strain against 7 sugarcane pathogenic fungi
Note that: the formula is antibacterial ratio (%) = (control group colony radius-treatment group colony radius)/control group colony radius×100;
The values in the table are mean value.+ -. Standard deviation;
* Represents a significant difference at the 1% level between the t-test post-treatment group and the control group.
Example 4 determination of the control effect of Bacillus bailii JB23 Strain on sugarcane root rot
(1) Preparation of Bacillus bailii JB23 biocontrol bacteria solution, inoculating Bacillus bailii JB23 strain stored in a refrigerator at-80 ℃ into 10mL of LB broth culture medium according to the ratio of 1:500, and shake culturing overnight at 37 ℃ and 220r/min to obtain seed solution. And inoculating the seed solution into LB broth culture medium, and culturing at 37 ℃ and 220r/min for 24 hours to obtain fresh biocontrol bacteria liquid. And centrifuging fresh JB23 bacterial liquid at 4000r/min for 20 minutes, pouring out waste liquid, re-suspending and precipitating with sterilized water, centrifuging at 4000r/min for 20 minutes again, pouring out waste liquid, adding sterilized water, and adjusting the OD 600 value of the strain to OD 600 =1 for later use.
(2) The preparation of the sugarcane root rot fungus spores, inoculating sugarcane root rot fungus BS46 stored in a refrigerator at the temperature of minus 80 ℃ into 10mL of PDW potato dextrose water culture medium according to the ratio of 1:500, carrying out shake cultivation for 48 hours at the temperature of 28 ℃ and 220r/min to obtain seed liquid, inoculating the seed liquid into PDW potato dextrose water, carrying out shake cultivation for 48 hours at the temperature of 28 ℃ and 220r/min to obtain fresh fungus spore liquid, filtering the fresh fungus liquid of BS46 by using 6 layers of sterilized gauze to obtain spores, centrifuging the filtrate at 4000r/min for 20 minutes, pouring out waste liquid, re-suspending precipitated spores by using sterilized water, centrifuging at 4000r/min for 20 minutes again, pouring out waste liquid, and regulating the spore number to 2X 10 6 CFU/mL by using sterilized water for standby.
(3) Potted plant test, selecting sugarcane variety badila susceptible to root rot, washing sugarcane stems with clear water to prepare single bud segments, then completely embedding the single bud sugarcane stems into perlite, rooting in dark at 28 ℃, digging out the sugarcane stems after 7 days, flushing the perlite attached to the surface with sterile water, soaking ① sugarcane stems in sterile water for 30 minutes, wherein the water control treatment is carried out; the treatment group ② soaks 15 cane stems with 2X 10 6 CFU/mL BS46 spore liquid prepared in advance for 30 minutes; the treatment group ③ is to soak 15 cane stems with 2× 6 CFU/mL BS46 spore liquid for 30 minutes, take out and naturally dry, soak JB23 biocontrol bacteria liquid with OD 600 =1 for 30 minutes, and the biocontrol bacteria treatment is performed. After air drying, all the sugarcane stems of the treatment group are completely embedded into the new perlite, and are cultivated in the dark for 10 days at 28 ℃. After 10 days, the surface-adherent perlite was excavated, rinsed with sterile water, and the growth was observed and recorded.
As shown in fig. 5 a-c, compared with the water treatment control of treatment group ①, the root of the sugarcane stem soaked in BS46 bacterial liquid in treatment group ② for 30 minutes has no new lateral root growth, the root growth also has obvious growth disorder and obvious blackish brown disease spots, the root of the sugarcane stem soaked in BS46 in treatment group ③ and then soaked in JB23 has new lateral root growth, and the root growth is not damaged obviously, and the result shows that bacillus beleisi JB23 can also play a good role in preventing and controlling BS46 in potting experiments.
Example 5 determination of the control effect of Bacillus bailii JB23 Strain on sugarcane tip rot
(1) The preparation of bacillus bailii JB23 biocontrol bacteria liquid was identical to the method mentioned in example 4 (1).
(2) The dominant strain CNO1 of the sugarcane top rot fungus spore is selected as an infection strain, and the specific spore preparation method refers to the preparation method of the sugarcane root rot fungus spore in the embodiment 4 (2).
(3) The potted plant test, selecting sugarcane No. I in sugarcane varieties susceptible to tip rot, cleaning sugarcane stems with clear water to prepare single bud segments, then completely embedding the single bud sugarcane stems into a matrix, and performing the following treatment on the sugarcane after 15 days of growth in the single pot matrix: ① 200. Mu.L of CNO1 spore solution at a concentration of 1X 10 6 CFU/mL was injected into the growing point, followed by 200. Mu.L of sterile water. ② 200. Mu.L of CNO1 spore liquid with the concentration of 1X 10 6 CFU/mL is injected into the growing point, and 200. Mu.L of JB23 bacterial liquid with OD 600 =1 is injected. ③ 200mL of JB23 bacterial liquid with OD 600 =1 is applied to the root of potted sugarcane 24 hours in advance, and 200 mu L of CNO1 spore liquid with concentration of 1X 10 6 CFU/mL is injected to a growing point after 24 hours. ④ 200. Mu.L of JB23 bacterial solution with OD 600 =1 was injected into the growing point, and 200. Mu.L of sterile water was further injected. ⑤ 400. Mu.L of sterile water was injected into the growth site. 15 replicates were made for each treatment. After one week, the number of plants and the grade of disease were observed and counted, and the results are shown in Table 3.
The morbidity calculation formula is: incidence (%) = number of disease plants/total number of investigation x 100;
disease index calculation formula disease severity index DSI (DISEASE SEVERITY index, DSI) = [ Σ (ni×vi)/4N ] ×100; where N represents the number of samples at the level, v represents the level, and N represents the total sample under test.
TABLE 3 statistics of the morbidity and index of disease for each treatment
As shown in table 3, JB23 can effectively and remarkably reduce the incidence rate and disease index of the tip rot caused by CNO1, and can play a good role in biocontrol.
Example 6 determination of the control Effect of Bacillus bailii JB23 Strain on sugarcane smut
(1) The preparation of bacillus bailii JB23 biocontrol bacteria liquid was identical to the method mentioned in example 4 (1).
(2) Preparing black spike spore powder, collecting black spike whips of sugarcane with smut in the field, then placing the black spike whips into an oven for drying to remove water, brushing down the black spike spore powder on the black spike whips by a brush after the black spike whips are dried, and collecting by a sampling bag for later use.
(3) Potted plant experiments, cutting a sugarcane variety ROC22 susceptible to sugarcane smut in the field, cutting the sugarcane variety ROC22 into single bud segments, and carrying out the following treatment before planting: treating ①, soaking sugarcane stems with black spike spore liquid for 30 minutes, taking out, airing, and soaking in sterile water for 30 minutes to serve as a control group; treating ②, diluting the black ear spore powder in the step (2) by using 2g of 1L of water, soaking the cane stems in the black ear spore powder liquid for 30 minutes, taking out, airing, and then soaking the biocontrol bacteria liquid for 30 minutes. 15 replicates were made for each treatment. Statistics were performed after continuous observation for 3 months, and the morbidity and the prevention effect were calculated according to the following formulas (1) and (2), and the results are shown in table 4.
Incidence = total number of lesions/total number of investigation x 100% (1)
Biocontrol effect= (control incidence-treatment incidence)/control incidence x 100% (2)
TABLE 4 statistics of incidence and biocontrol Effect of sugarcane smut treatments
| Treatment of | Incidence of disease | Biocontrol effect |
| Treatment of ① | 13.6％ | —— |
| Treatment of ② | 0 | 100％ |
As can be seen from table 4, the incidence of JB23 treated sugarcane was 0 after three months of inoculation. The JB23 strain has very good effect of preventing and controlling black spikes in the field and has obvious biocontrol effect.
In conclusion, the bacillus bailii JB23 strain provided by the invention has remarkable inhibition effect on the growth of various sugarcane pathogenic fungi, can be used for biological control of sugarcane fungal diseases, and provides new strain resources for a sugarcane disease biocontrol strain resource library. In addition, the bacillus belicus JB23 strain can reduce the incidence rate of root rot, tip rot and smut, reduce the damage of diseases to the root of sugarcane, improve the control effect and have wide development prospect.
The above embodiments are only illustrative of the preferred embodiments of the present invention and are not intended to limit the scope of the present invention, and various modifications and improvements made by those skilled in the art to the technical solutions of the present invention should fall within the protection scope defined by the claims of the present invention without departing from the design spirit of the present invention.
Claims (7)
1. The bacillus belgium (Bacillus velezensis) JB23 strain is characterized in that the strain is preserved in China center for type culture collection, the preservation time is 2022, 7 months and 18 days, the preservation address is China center for type culture collection, the university of Wuhan in Wuhan district, wuhan, hubei province, and the preservation number is CCTCC NO: m20221134.
2. A microbial agent comprising the Bacillus bailii JB23 strain or a fermented bacterial liquid thereof according to claim 1.
3. The microbial inoculum of claim 2, wherein the concentration of bacillus beliensis JB23 in the fermentation broth is equal to or greater than 10 8 cfu/mL.
4. Use of the bacillus belgium JB23 strain of claim 1 or the microbial inoculum of any one of claims 2-3 for inhibiting the growth of a sugarcane pathogenic fungus, wherein the sugarcane pathogenic fungus is one or more of a sugarcane root rot fungus, a sugarcane tip rot fungus, and a sugarcane smut fungus.
5. The use according to claim 4, wherein the sugarcane root rot fungus comprises fusarium shared; the fusarium graminearum comprises fusarium graminearum, fusarium neoformans, fusarium moniliforme and fusarium oxysporum; the sugarcane smut bacteria comprise sugarcane smut bacteria.
6. The bacillus bailii JB23 strain of claim 1 or the use of the microbial inoculum of any one of claims 2-3 in the manufacture of a medicament for controlling sugarcane fungal diseases, wherein the sugarcane fungal diseases include diseases caused by infection of sugarcane roots, stems, leaves with sugarcane pathogenic fungi;
The sugarcane pathogenic fungi are one or more of sugarcane root rot fungi, sugarcane tip rot fungi and sugarcane smut fungi.
7. A method for controlling sugarcane fungal disease, characterized in that bacillus belicus JB23 strain according to claim 1 or the microbial inoculum according to any one of claims 2-3 is applied to sugarcane roots, stems, leaves;
The sugarcane pathogenic fungi are one or more of sugarcane root rot fungi, sugarcane tip rot fungi and sugarcane smut fungi.
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