CN114703094B - Bacillus pseudomycosis and application thereof in preparation of plant pathogenic bacteria bacteriostat - Google Patents
Bacillus pseudomycosis and application thereof in preparation of plant pathogenic bacteria bacteriostat Download PDFInfo
- Publication number
- CN114703094B CN114703094B CN202210272700.0A CN202210272700A CN114703094B CN 114703094 B CN114703094 B CN 114703094B CN 202210272700 A CN202210272700 A CN 202210272700A CN 114703094 B CN114703094 B CN 114703094B
- Authority
- CN
- China
- Prior art keywords
- bacillus
- pseudomycosis
- fusarium
- pathogenic bacteria
- plant pathogenic
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Classifications
-
- 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
Abstract
The invention belongs to the technical field of microorganisms, and particularly relates to a bacillus pseudomycosis FJH2 and application thereof in preparation of plant pathogenic bacteria bacteriostat, wherein the bacillus pseudomycosis FJH2 has a preservation number of CCTCC NO: M2022049. The strain has the functions of resisting plant pathogenic bacteria and the like; can grow in the salt concentration of 1% -6% at the pH of 6.0-8.0 and at the temperature of 20-40 ℃, has good inhibition effect on several plant pathogenic bacteria, has the inhibition rate on fusarium oxysporum up to 91.47%, and has potential application value in preparing biocontrol bacteria.
Description
Technical Field
The invention belongs to the technical field of microorganisms, and particularly relates to bacillus pseudomycosis and application thereof in preparation of plant pathogenic bacteria bacteriostat.
Background
Fusarium is widely existing in soil and plants, and mainly acts on the junctions of roots and rhizomes of plants to destroy vascular bundle tissues of the plants, and in the metabolic process, metabolites such as enzymes, phenols and the like are produced to influence the growth and development of host plants. Plant wilt occurs most seriously in summer, and infected plants mainly show slow development, yellow leaves, dead wilt fallen leaves, rotten rhizome and the like. At present, no chemical pesticide which is very effective in treating the fusarium wilt exists in agriculture, so that the fusarium wilt treatment mainly depends on a biological control method. Studies show that the bacillus pseudomycosis can produce plasmin to increase the potassium absorption of crops, but few reports on bacteriostasis of the bacillus pseudomycosis are provided.
The invention screens out a strain of bacillus pseudomycosis, which has good inhibition effect on fusarium oxysporum and fusarium solani, and has very high inhibition rate, and the inhibition rate on fusarium oxysporum reaches 91.47%; can play a certain role in preventing and controlling plant diseases, and has great potential for developing biocontrol bacteria.
Disclosure of Invention
The invention aims to provide a strain of bacillus pseudomycoides (FJH 2), wherein the preservation number of the bacteria is as follows: cctccc No. M2022049.
Another object of the invention is to provide the use of a strain of bacillus pseudomycosis for the preparation of a plant pathogenic bacteria inhibitor.
In order to achieve the above object, the present invention adopts the following technical measures:
the applicant separates a strain of bacillus pseudomycosis from soil samples of the city of Pucheng county in the Wei south of Shaanxi, has antibacterial effect on pathogenic bacteria such as fusarium oxysporum, fusarium solani and the like, has higher antibacterial rate, is identified as bacillus pseudomycosis, and is sent to China center for type culture collection for preservation in the 1 st month of 2022, and the address is the university of Wuhan in China, which is named after classification: bacillus pseudomycoides FJH2, accession number: cctccc No. M2022049.
The colony morphology of the isolated Bacillus pseudomycosis (Bacillus pseudomycoides) on LB solid medium is shown in FIG. 1, and the colony morphology is in the form of off-white and flat root. The physiological and biochemical performance is gram-positive, non-hydrolyzable starch, non-hydrolyzable cellulose, negative in methyl red test, negative in acetyl methyl methanol test and positive in catalase.
The application of the bacillus pseudomycosis FJH2 in preparing a plant pathogenic bacteria bacteriostatic agent, wherein the plant pathogenic bacteria are banana Fusarium wilt (Fusarium oxysporum F.sp.cube), fusarium oxysporum (Fusarium oxysporum), penicillium brasiliensis (Penicillium brasilianum), fusarium solani (Fusarium solani), fusarium chlamydosporium (Fusarium chlamydosporum), penicillium (Penicillium glaucum) and Mucor (Mucor).
Compared with the prior art, the invention has the following characteristics:
the invention discloses a bacillus pseudomycosis for the first time, which has a good antibacterial effect on a plurality of pathogenic bacteria causing plant wilt.
Drawings
FIG. 1 is a diagram showing the culture characteristics of Bacillus pseudomycosis FJH2.
FIG. 2 is a schematic representation of the inhibition of Fusarium oxysporum by Bacillus pseudomycoides FJH2.
FIG. 3 is a schematic representation of the inhibition of Fusarium solani by Bacillus pseudomycoides FJH2.
FIG. 4 is a schematic representation of the inhibition of Mucor by Bacillus pseudomycosis FJH2.
Detailed Description
The technical scheme of the invention is conventional technology unless specified; the reagents or materials, unless otherwise specified, are commercially available.
Example 1:
isolation and identification of Bacillus pseudomycosis:
2.5g of soil sample from Puchen county, weisouth Shaanxi is placed in 50ml of sterile water, shaken at 30℃for 1h at 250rpm, after which 500. Mu.l of soil suspension is taken for gradient dilution in a PA bottle containing 4.5ml of sterile water. Take 10 -1 、10 -2 、10 -3 、10 -4 、10 -5 The soil suspensions with the dilutions are respectively added into a culture medium for coating, the growth conditions of the plate colonies are observed after the culture for 24 hours, 48 hours, 72 hours and 96 hours, all the grown colonies are respectively picked out, and an antibacterial activity experiment is carried out; selecting bacteria with high antibacterial rate and broad antibacterial spectrum; the Bacillus pseudomycosis FJH2 was finally selected.
(1) Morphological features
The morphology of the isolated Bacillus pseudomycosis colonies on LB solid medium is shown in FIG. 1, and the colonies are in the shape of white and flat roots.
(2) Physiological and biochemical experiments
The physiological and biochemical performance shows that gram staining shows positive, non-hydrolytic starch, non-hydrolytic cellulose, methyl red test negative, acetyl methyl methanol test negative and catalase positive.
(3) Gene identification of Bacillus pseudomycosis
Extracting genome DNA of the strain to be detected by using a bacterial DNA extraction kit, wherein the specific operation steps are referred to the instruction book. Using the extracted bacterial genome DNA as a template, amplifying bacteria, and amplifying a primer of 16S rRNA:
27F:5＇-gtttgatcctggctcag-3＇
1492R:5＇-tacggctaccttgttacgactt-3＇；
the sequence is subjected to homology comparison analysis by Blast16SrDNA sequence, and the result analysis shows that the sequence has the highest sequence similarity with the 16SrDNA gene of the strain Bacillus pseudomycoide, belongs to a bacillus pseudomycosis, and is sent to China center for type culture collection for preservation in 2022, and addresses are as follows: bacillus pseudomycoides FJH2, accession number: cctccc No. M2022049.
Example 2:
tolerance test of bacillus pseudomycosis:
(1) pH resistance Range experiment:
inoculating the seed solution of Bacillus pseudomycosis into LB culture medium with different initial pH according to 2%, culturing at 30deg.C under 200r/min for 16 hr, sampling, and measuring OD 600 . The initial pH gradient of the medium was set to 3, 4, 5, 6, 7, 8, 9, 10 (initial 0D was 0).
LB medium: 10g of peptone, 5g of yeast extract powder, 10g of NaCl, 1000mL of distilled water, pH7.0 and high-pressure steam sterilization at 121 ℃ for 20min.
| PH | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 |
| OD 600 | 0 | 0 | 0 | 4.712 | 5.356 | 4.504 | 0 | 0 |
(2) Salt tolerance concentration test:
inoculating the seed solution of Bacillus pseudomycosis into LB culture medium with different salt concentrations according to 2%, culturing at 30deg.C under 200r/min for 16 hr, sampling, and measuring OD 600 . The salt concentration gradient of the medium was set to 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10% (initial 0D is 0).
LB medium: 10g of peptone, 5g of yeast extract powder, 10g of NaCl, 1000mL of distilled water, pH7.0 and high-pressure steam sterilization at 121 ℃ for 20min.
| Salt concentration | 1％ | 2％ | 3％ | 4％ | 5％ | 6％ | 7％ | 8％ |
| OD 600 | 3.298 | 3.51 | 4.5 | 3.122 | 2.248 | 1.516 | 0 | 0 |
(3) Growth temperature experiment:
inoculating the seed solution of Bacillus pseudomycosis in LB culture medium at 2%, shake culturing at 200r/min under different temperature conditions for 16 hr, sampling, and measuring OD 600 . The temperature gradient of the medium was set at 20 ℃, 28 ℃, 37 ℃, 40 ℃, 45 ℃ (initial 0D was 0).
| Temperature (. Degree. C.) | 20℃ | 25℃ | 30℃ | 35℃ | 40℃ | 45℃ |
| OD600 | 0.3 | 2.728 | 6.28 | 5.176 | 4.892 | 0 |
Example 3:
inhibition of plant pathogens by bacillus pseudomycosis:
the inhibition capacity of the bacillus pseudomycosis to pathogenic bacteria is measured by adopting a bacteriostasis circle method. The Bacillus pseudomycosis was activated with LB, and the seed solution was appropriately diluted and applied to PDA plates to apply sterile water as a control test group. Punching on PDA plate full of pathogenic fungi with gun head, inoculating fungus block onto PDA plate coated with Bacillus pseudomycosis, culturing at 37deg.C for 4d, measuring fungus colony diameter, calculating antibacterial rate, and analyzing antibacterial effect.
The plant pathogenic bacteria selected in this experiment were banana Fusarium oxysporum f.sp.cube, fusarium oxysporum (Fusarium oxysporum), penicillium brazil (Penicillium brasilianum), fusarium solani (Fusarium solani), fusarium chlamydosporium (Fusarium chlamydosporium), penicillium (Penicillium glaucum) and Mucor (Mucor).
PDA medium: 200g of potato, 20g of glucose, 1000mL of distilled water, and sterilizing by high-pressure steam at 115 ℃ for 30min, wherein the pH is natural.
Note that: d, control plate colony diameter (mm); d, experimental plate colony diameter (mm).
The bacillus pseudomycosis has very good control effect on fusarium oxysporum, and the inhibition rate on pathogenic bacteria causing the plant diseases reaches 91.47 percent; has a certain inhibition effect on fusarium solani and mucor, and the inhibition rate reaches 69.48 percent and 59.76 percent. The inhibition effect on penicillium, fusarium chlamydophim, fusarium oxysporum and Fusarium putrescens is poor. The bacteriostasis rate for each bacterium is shown in the following table.
From the above examples, the present invention provides a strain of Bacillus pseudomycosis FJH2 having a good control effect for controlling plant blight caused by Fusarium oxysporum and Fusarium solani.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.
Sequence listing
<110> Shandong wild field biotechnology Co., ltd
<120> A strain of Bacillus pseudomycosis and use thereof in preparing plant pathogenic bacteria inhibitor
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 17
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 1
gtttgatcct ggctcag 17
<210> 2
<211> 22
<212> DNA
<213> Artificial sequence (Artificial Sequence)
<400> 2
tacggctacc ttgttacgac tt 22
Claims (2)
1. Isolated bacillus pseudomycosisBacillus pseudomycoides) The preservation number of the bacillus pseudomycosis is as follows: cctccc No. M2022049.
2. The use of a pseudomycobacillus of claim 1 in the preparation of a plant pathogenic bacteria inhibitor, said plant pathogenic bacteria being banana vascular wilt bacteria @Fusarium oxysporum F.sp. Cubense) Fusarium oxysporum (F.oxysporum)Fusarium oxysporum) Penicillium BrazilPenicillium brasilianum) Fusarium solani (Fusarium solani)Fusarium solani) Fusarium chlamydiaFusarium chlamydosporum) Penicillium glaucum (L.) KuntzePenicillium glaucum) Mucor mouldMucor）。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210272700.0A CN114703094B (en) | 2022-03-18 | 2022-03-18 | Bacillus pseudomycosis and application thereof in preparation of plant pathogenic bacteria bacteriostat |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202210272700.0A CN114703094B (en) | 2022-03-18 | 2022-03-18 | Bacillus pseudomycosis and application thereof in preparation of plant pathogenic bacteria bacteriostat |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN114703094A CN114703094A (en) | 2022-07-05 |
| CN114703094B true CN114703094B (en) | 2023-09-05 |
Family
ID=82168829
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202210272700.0A Active CN114703094B (en) | 2022-03-18 | 2022-03-18 | Bacillus pseudomycosis and application thereof in preparation of plant pathogenic bacteria bacteriostat |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN114703094B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117431191B (en) * | 2023-12-19 | 2024-03-12 | 中国农业大学 | Bacillus pseudomycoides HDCP1 and application thereof |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2307010A1 (en) * | 2000-05-19 | 2001-11-19 | Infectio Diagnostic (I.D.I.) Inc. | Highly conserved genes and their use to generate species-specific, genus-specific, family-specific, group-specific and universal nucleic acid probes and amplification primers to rapidly detect and identify bacterial,fungal and parasitical pathogens from clinical specimens for diagnosis |
| CN103619174A (en) * | 2011-05-13 | 2014-03-05 | Isp投资公司 | Aqueous solutions of 1,2-benzisothiazolin-3-one |
| WO2017189589A1 (en) * | 2016-04-25 | 2017-11-02 | Isp Investments Llc | An antimicrobial composition, process for preparing the same and method of use thereof |
| CN110218682A (en) * | 2019-06-26 | 2019-09-10 | 南京农业大学 | One plant of pseudomycete sample bacillus and its application in mud decrement |
| CN113736679A (en) * | 2020-05-30 | 2021-12-03 | 华中农业大学 | Cold-resistant bacillus FJW2 and application thereof in preparation of plant pathogenic bacteria bacteriostat |
| CN115960777A (en) * | 2022-12-06 | 2023-04-14 | 江苏省中国科学院植物研究所 | Pseudomycobacillus sp CNBG-PGPR-20 and application thereof in prevention and treatment of vegetable epidemic diseases |
| CN116083297A (en) * | 2022-12-02 | 2023-05-09 | 洛阳理工学院 | Microbial agent for preventing and treating strawberry root rot and preparation method thereof |
-
2022
- 2022-03-18 CN CN202210272700.0A patent/CN114703094B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2307010A1 (en) * | 2000-05-19 | 2001-11-19 | Infectio Diagnostic (I.D.I.) Inc. | Highly conserved genes and their use to generate species-specific, genus-specific, family-specific, group-specific and universal nucleic acid probes and amplification primers to rapidly detect and identify bacterial,fungal and parasitical pathogens from clinical specimens for diagnosis |
| CN103619174A (en) * | 2011-05-13 | 2014-03-05 | Isp投资公司 | Aqueous solutions of 1,2-benzisothiazolin-3-one |
| WO2017189589A1 (en) * | 2016-04-25 | 2017-11-02 | Isp Investments Llc | An antimicrobial composition, process for preparing the same and method of use thereof |
| CN110218682A (en) * | 2019-06-26 | 2019-09-10 | 南京农业大学 | One plant of pseudomycete sample bacillus and its application in mud decrement |
| CN113736679A (en) * | 2020-05-30 | 2021-12-03 | 华中农业大学 | Cold-resistant bacillus FJW2 and application thereof in preparation of plant pathogenic bacteria bacteriostat |
| CN116083297A (en) * | 2022-12-02 | 2023-05-09 | 洛阳理工学院 | Microbial agent for preventing and treating strawberry root rot and preparation method thereof |
| CN115960777A (en) * | 2022-12-06 | 2023-04-14 | 江苏省中国科学院植物研究所 | Pseudomycobacillus sp CNBG-PGPR-20 and application thereof in prevention and treatment of vegetable epidemic diseases |
Non-Patent Citations (1)
| Title |
|---|
| 黄连根腐病病原菌的分离鉴定及其拮抗菌筛选;廖海浪等;中国抗生素杂志;第47卷(第9期);第900-912页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN114703094A (en) | 2022-07-05 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108998389B (en) | Pseudomonas having antagonistic action on xanthomonas oryzae and magnaporthe grisea and application thereof | |
| CN109456921B (en) | Paenibacillus polymyxa, application thereof, microbial agent, powder and granules | |
| CN107099478A (en) | One plant of Lycoris aurea endogenetic bacteria bulkholderia cepasea HDXY 02 and its application | |
| CN106167776A (en) | A kind of can bacillus cereus (Bacillus cereus) TH 35 of heavy metal cadmium and application thereof in activating soil | |
| CN112662585B (en) | Bacillus atrophaeus DX-9 and application thereof | |
| CN113201478B (en) | Siamese bacillus Gxun-36 secreting iron carrier and application thereof | |
| CN106190892B (en) | One bacillus subtilis strain and its application | |
| CN114703094B (en) | Bacillus pseudomycosis and application thereof in preparation of plant pathogenic bacteria bacteriostat | |
| CN113249243A (en) | Pseudomonas, microbial biocontrol microbial inoculum containing pseudomonas and application of microbial biocontrol microbial inoculum | |
| CN110184224B (en) | Staphylococcus epidermidis and application thereof | |
| CN112011478A (en) | Dendrobium nobile endogenous Burkholderia gladioli BL-HTie-5 and application thereof | |
| CN113980876A (en) | Bacillus belgii and application thereof | |
| CN112592856B (en) | Tobacco brown spot antagonistic actinomycete strain and application thereof | |
| CN107629985B (en) | Plant endophytic bacterium with antagonistic effect on plant pathogenic fungi | |
| CN110452859B (en) | Bacillus amyloliquefaciens B4216 and application thereof | |
| CN105112324B (en) | The stench Marseille bacterium of one kind production dimethyl disulfide (DMDS) and its application | |
| CN113736679B (en) | Cold-resistant bacillus FJW2 and application thereof in preparation of plant pathogenic bacteria bacteriostatic agent | |
| CN107058183B (en) | Bacillus methylotrophicus, and biocontrol microbial inoculum and application thereof | |
| CN112760229B (en) | Trichoderma and application thereof | |
| CN110484463B (en) | Bacillus mojavensis B282 and application thereof | |
| CN112391312B (en) | Streptomyces and application thereof in prevention and treatment of plant oomycete diseases | |
| CN111662843B (en) | Bacillus amyloliquefaciens and separation and screening method and application thereof | |
| CN114250162B (en) | Bacillus firmus and application thereof in preparation of plant pathogenic bacteria bacteriostatic agent | |
| CN111793571A (en) | Saline-alkali-tolerant bacillus megaterium FJW1 and application thereof in preparation of plant pathogenic bacteria bacteriostat | |
| CN114149949B (en) | Streptomyces JXJA01 and biological agent and application thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| CB03 | Change of inventor or designer information |
Inventor after: Huang Li Inventor after: Su Shanzhu Inventor after: Yang Shaozhou Inventor after: Liu Yonggao Inventor after: Ding Xiangqi Inventor after: Chen Zhenmin Inventor after: Yan Xunbo Inventor after: Zhu Qiuyan Inventor after: Qi Hongyue Inventor after: Feng Ke Inventor after: Geng Yuefeng Inventor after: Zhang Xinlong Inventor after: Jia Zhongqing Inventor before: Huang Li Inventor before: Su Shanzhu Inventor before: Yang Shaozhou Inventor before: Liu Yonggao Inventor before: Ding Xiangqi Inventor before: Chen Zhenmin Inventor before: Yan Xunbo Inventor before: Zhu Qiuyan Inventor before: Qi Hongyue Inventor before: Feng Ke Inventor before: Geng Yuefeng Inventor before: Zhang Xinlong Inventor before: Jia Zhongqing |
|
| CB03 | Change of inventor or designer information | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |