CN110408548A - It is a kind of prevent and treat Pattern of Asarum Leaf Blight Antagonistic Fungus bacterial strain and its application - Google Patents
It is a kind of prevent and treat Pattern of Asarum Leaf Blight Antagonistic Fungus bacterial strain and its application Download PDFInfo
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- CN110408548A CN110408548A CN201910742190.7A CN201910742190A CN110408548A CN 110408548 A CN110408548 A CN 110408548A CN 201910742190 A CN201910742190 A CN 201910742190A CN 110408548 A CN110408548 A CN 110408548A
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Abstract
The invention discloses a kind of Antagonistic Fungus bacterial strain for preventing and treating Pattern of Asarum Leaf Blight and its applications, the Antagonistic Fungus bacterial strain is isolated from healthy asarum plant rhizosphere soil, for antagonism Pattern of Asarum Leaf Blight pathogen, classification naming are as follows: microassembly robot (Penicillium janthinellum);Deposit number: CGMCC No.18101;The present invention from asarum healthy plant rhizosphere soil by isolating the Antagonistic Fungus bacterial strain-penicillium janthinellum for having biocontrol effect to Pattern of Asarum Leaf Blight, then the test of living body control efficiency is carried out to the Antagonistic Fungus bacterial strain filtered out, the prevention preventive effect for obtaining Antagonistic Fungus bacterial strain is 70.78%, treating preventive effect is 61.03%, Antagonistic Fungus bacterial strain has preferable effect in terms of controlling state of an illness extension after prevention Pattern of Asarum Leaf Blight occurs and occurs, the raising that Disease Resistance-Related Enzymes in asarum plant body can be induced, can promote the development of Pattern of Asarum Leaf Blight biological control.
Description
Technical field
The present invention relates to microbiology engineering and technical field more particularly to a kind of Antagonistic Fungus for preventing and treating Pattern of Asarum Leaf Blight
Bacterial strain and its application.
Background technique
Asarum also known as thin ginseng, bowl of a pipe flower, are pungent the categorys perennial grass undergraduate course plant of Aristolochiaceae, are important medicinal of one kind
Plant, asarum is warm-natured, acrid flavour, has warming channel and expelling cold, wind-expelling pain-stopping, sensible, relieving cough and asthma, promoting blood circulation effect, for treating
The diseases such as anemofrigid headache, phlegm retention cough and asthma, arthralgia, dentalgia, nasosinusitis.Pattern of Asarum Leaf Blight is typical polycyclic disease, pathogen
For M. acerina (Mycocentrospora acerina (Hartig) Deihton), conidium is main source of infection, can
It is propagated by splash gentle stream of rainwater, main harm blade, scab subcircular, with wheel line around sepia, serious disease occurs
Mottled rot is rotten, and perforation, entire blade is withered, seriously affects the yield and quality of asarum.
Traditional chemical control method has the features such as quick, wide sterilization spectrum, but caused by the unreasonable use of chemical pesticide
The problems such as medicament residue, food safety and environmental pollution, increasingly shows, and even results in pest resurgence and disease drug resistance is passed
Phenomena such as increasing, and biological control can prevent and treat plant pest under conditions of free from environmental pollution, have safety, noresidue, specifically
Property it is strong the advantages that, using biocontrol bacterial strain prevention and treatment asarum disease facilitate medicinal material quality, improve medicinal material yield, have ecology and
Economic double benefit, less to the research of Pattern of Asarum Leaf Blight biological control both at home and abroad at present, chemical pesticide is to Pattern of Asarum Leaf Blight
Prevention and treatment will cause medicament residue to influence the quality and quality of medicinal material, therefore, the present invention proposes that a kind of prevention and treatment asarum leaf is withered
The Antagonistic Fungus bacterial strain of disease and its application, to solve shortcoming in the prior art.
Summary of the invention
In view of the above-mentioned problems, the present invention propose it is a kind of prevent and treat Pattern of Asarum Leaf Blight Antagonistic Fungus bacterial strain and its application, pass through
Antagonistic Fungus bacterial strain-microassembly robot that there is biocontrol effect to Pattern of Asarum Leaf Blight is isolated from asarum healthy plant rhizosphere soil
Then bacterium carries out bacteriostatic test and living body control efficiency in plate to the Antagonistic Fungus bacterial strain filtered out and tests, obtained antagonism
The prevention preventive effect of fungal bacterial strain is 70.78%, and treatment preventive effect is 61.03%, and Antagonistic Fungus bacterial strain is in prevention Pattern of Asarum Leaf Blight hair
Control state of an illness extension aspect has preferable effect after raw and generation, and can induce disease resistance related enzymes in asarum plant body
Active raising can effectively facilitate the development of Pattern of Asarum Leaf Blight biological control.
The present invention proposes a kind of Antagonistic Fungus bacterial strain for preventing and treating Pattern of Asarum Leaf Blight, and the Antagonistic Fungus bacterial strain is from healthy asarum
It is isolated in plant rhizosphere soil, it is used for antagonism Pattern of Asarum Leaf Blight pathogen;Classification naming are as follows: microassembly robot
(Penicillium janthinellum);Deposit number: CGMCC No.18101;The preservation time: on June 21st, 2019;It protects
Hide address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica;Depositary institution: the micro- life of China
Object culture presevation administration committee common micro-organisms center.
Further improvement lies in that: the filtering of the strain culture, fermentation liquid or fermentation culture of the Antagonistic Fungus bacterial strain
Liquid.
Further improvement lies in that: the Antagonistic Fungus bacterial strain is the bacteria agent of active constituent in Pattern of Asarum Leaf Blight prevention and treatment
Application.
Further improvement lies in that: the occurrence and development of Antagonistic Fungus bacterial strain control Pattern of Asarum Leaf Blight, have prevention and
Therapeutic effect.
It is a kind of prevent and treat Pattern of Asarum Leaf Blight Antagonistic Fungus bacterial strain prevention and treatment the application of pathogen-M. acerina.
The invention has the benefit that the method for the present invention from asarum healthy plant rhizosphere soil by isolating to asarum
There is leaf blight Antagonistic Fungus bacterial strain-penicillium janthinellum of biocontrol effect to pass through plate pair using Pattern of Asarum Leaf Blight bacterium as target bacterium
Method of standing erect screens Antagonistic Fungus bacterial strain, then carries out the test of living body control efficiency to the Antagonistic Fungus bacterial strain filtered out, obtains
The prevention preventive effect for having gone out Antagonistic Fungus bacterial strain is 70.78%, and treatment preventive effect is 61.03%, and Antagonistic Fungus bacterial strain is in prevention asarum
Control state of an illness extension aspect has preferable effect after leaf blight occurs and occurs, and can induce disease-resistant in asarum plant body
Property related enzyme activity raising provide science for the biological control prevention and treatment of Pattern of Asarum Leaf Blight to resist infecting for disease
Data are supported, the development of Pattern of Asarum Leaf Blight biological control can be effectively facilitated.
Detailed description of the invention
Fig. 1 is that constant temperature opposite culture of the present invention inhibits Pattern of Asarum Leaf Blight pathogen effect diagram.
Fig. 2 is the hypha form schematic diagram of Pattern of Asarum Leaf Blight pathogen after Antagonistic Fungus bacterial strain of the present invention processing.
Fig. 3 is Antagonistic Fungus bacterial strain Immune effect schematic diagram of the present invention.
Fig. 4 is that Antagonistic Fungus bacterial strain of the present invention treats effect of inoculation schematic diagram.
Fig. 5 is the active schematic diagram of polyphenol oxidase (PPO) in Antagonistic Fungus bacterial strain inducing asarum plant body of the present invention.
Fig. 6 is the active schematic diagram of phenylalanine lyase (PAL) in Antagonistic Fungus bacterial strain inducing asarum plant body of the present invention.
Fig. 7 is the active schematic diagram of peroxidase (POD) in Antagonistic Fungus bacterial strain inducing asarum plant body of the present invention.
Fig. 8 is the active schematic diagram of superoxide dismutase (SOD) in Antagonistic Fungus bacterial strain inducing asarum plant body of the present invention.
Specific embodiment
In order to deepen the understanding of the present invention, the present invention is further described below in conjunction with embodiment, the present embodiment
For explaining only the invention, it is not intended to limit the scope of the present invention..
Embodiment one
The present embodiment proposes Antagonistic Fungus bacterial strain-penicillium janthinellum separation screening and identification: from asarum healthy plant root
A variety of bacterial strains are isolated by gradient dilution method in the soil of border, using Pattern of Asarum Leaf Blight bacterium as target bacterium, using the side of plate face-off
Method filters out the Antagonistic Fungus bacterial strain that one plant of number is F1-6, and through morphology and molecular biology identification, classification naming is micro-
Purple Penicillium notatum (Penicillium janthinellum);
The separation screening detailed process of Antagonistic Fungus bacterial strain the following steps are included:
Step 1: Antagonistic Fungus bacterial strain is separated using soil dilution partition method, 10g asarum healthy plant rhizosphere soil is taken to add
Enter into the triangular flask of 90mL sterile water, triangular flask is then subjected to concussion processing 20min, the speed for shaking processing is 160r/
Min is mixed evenly to prepare soil supension, then soil supension is diluted to 10-4、10-5、10-6Three gradients;
Step 2: respectively from 10-4、10-5、10-6100uL even spread is drawn in the dilution soil supension of three gradients to arrive
In PDA plate;Each gradient is coated with 3 plates as repeating, obtain 9 even spreads the PDA plate of dilution soil supension;
Step 4: the PDA plate of 9 even spreads dilution soil supension is subjected to constant temperature incubation, constant temperature incubation temperature
It is 22 DEG C, the constant temperature incubation time is 2 days, is purified according to colonial morphology, by obtained variety classes bacterial strain in 4 DEG C of asepsis rings
It is saved backup under border;
Step 5: using Pattern of Asarum Leaf Blight bacterium as target bacterium, Antagonistic Fungi is screened using the method for plate face-off, it is true to obtain antagonism
Then bacteria strain carries out strain idenfication further according to the method for morphology and molecular biology, qualification result indicates, Antagonistic Fungus bacterium
Strain culture forms more typical bacterium colony for 4 days, is in yellow among bacterium colony, and edge is white, diameter 20-30mm, regular edges,
Bacterium colony surface is cotton-shaped;
ITS sequence analysis is carried out to Antagonistic Fungus bacterial strain, nucleic acid sequence BLAST analysis is carried out in GenBank database,
The result shows that the homology of Antagonistic Fungus bacterial strain and Penicillium janthinellum are 99%, it is determined as penicillium janthinellum
(Penicillium janthinellum)。
Embodiment two
According to Fig. 1,2, the present embodiment proposes Antagonistic Fungus bacterial strain-inhibition of the penicillium janthinellum to Pattern of Asarum Leaf Blight bacterium
Effect: carrying out constant temperature opposite culture for Antagonistic Fungus bacterial strain-penicillium janthinellum and Pattern of Asarum Leaf Blight pathogen-M. acerina, permanent
Warm opposite culture temperature is 25 DEG C, and the constant temperature incubation time is 7 days, and observes the pathogen edge hypha form of opposite culture;
Through Antagonistic Fungus bacterial strain-penicillium janthinellum treated Pattern of Asarum Leaf Blight pathogen-M. acerina (such as Fig. 1 institute
Show), under the microscope it has been observed that pathogen colony edge mycelia often has greatly fusing, contracting of hanging, chlamydospore are increased, content
The phenomenon that object is reduced (as shown in Figure 2), shows Antagonistic Fungus bacterial strain-penicillium janthinellum to Pattern of Asarum Leaf Blight pathogen-maple bacteria thorn
The growth of spore significantly inhibits.
Embodiment three
According to Fig. 3,4, the present embodiment proposes that Antagonistic Fungus bacterial strain-penicillium janthinellum prevents and treats Pattern of Asarum Leaf Blight potting
Effect measuring, comprising the following steps:
Step 1: control effect testing is divided into 3 processing, respectively at immunization campaign Antagonistic Fungus bacterial strain-penicillium janthinellum
Reason group, treatment inoculation Antagonistic Fungus bacterial strain-penicillium janthinellum processing group and only inoculation leaf blight pathogen-M. acerina sprinkling
Sterile water as a control group, then chooses the consistent asarum blade of 300 growing ways, each processing group from the potting asarum of health
100, and 6 circular holes are respectively pricked on each blade;
Step 2: Antagonistic Fungus bacterial strain-penicillium janthinellum spore is rinsed with sterile water, being configured to concentration is 1 × 108
The spore suspension of a/mL;
Step 3: immunization campaign processing group first uniformly sprays spore suspension, is inoculated with Pattern of Asarum Leaf Blight pathogen-maple after 3h
Bacteria thorn spore, treatment inoculation processing group are first inoculated with Pattern of Asarum Leaf Blight pathogen-M. acerina, in uniformly spraying spore on blade after 48h
Sub- suspension, to spray sterile water as control group (CK group), moisturizing culture started to observe incidence after 3 days, carried out prevention and treatment effect
Fruit statistics, morbidity classification standard is as shown in table 1, shown in disease index calculation formula such as formula (1), preventive effect calculation formula such as formula
(2) shown in:
The morbidity classification standard of table 1
Grade | Standard |
0 grade | Disease-free spot |
1 grade | Lesion area accounts for 5% or less entire leaf area |
3 grades | Lesion area accounts for the 6%-10% of entire leaf area |
5 grades | Lesion area accounts for the 11%-25% of entire leaf area |
7 grades | Lesion area accounts for the 26%-50% of entire leaf area |
9 grades | Lesion area accounts for 50% or more of entire leaf area |
Show only to be inoculated with Pattern of Asarum Leaf Blight pathogen-M. acerina processing group blade disease after carrying out experiment of preventive effects
Feelings index is 30.67, and Antagonistic Fungus bacterial strain-penicillium janthinellum processing disease index is 8.96, and prevention preventive effect is 70.78%
(as shown in Figure 3);
Show only to be inoculated with Pattern of Asarum Leaf Blight pathogen-M. acerina processing group blade disease after carrying out therapeutic effect test
Feelings index is 78.29, and Antagonistic Fungus bacterial strain-penicillium janthinellum processing disease index is 30.51, and treatment preventive effect is
61.03% (as shown in Figure 4);
Compare Antagonistic Fungus bacterial strain-penicillium janthinellum to the prevention and treatment effect test result of Pattern of Asarum Leaf Blight pathogen
Show that Antagonistic Fungus bacterial strain-penicillium janthinellum prevention preventive effect is 70.78%, treatment preventive effect is 61.03%, Antagonistic Fungus bacterium
Strain-penicillium janthinellum has preferable effect in terms of controlling state of an illness extension after prevention Pattern of Asarum Leaf Blight occurs and occurs.
Example IV
According to Fig. 5,6,7,8, the present embodiment proposes that Antagonistic Fungus bacterial strain-penicillium janthinellum resists in asarum plant body
The measurement of characteristic of disease related enzyme activity: the 0th, 2,4,6,8 day progress plant disease-resistant relevant enzyme after immunization campaign and treatment inoculation processing
Active measurement, including to polyphenol oxidase (PPO), phenylalanine lyase (PAL), peroxidase (POD) and super oxygen
The enzyme assay of object mutase (SOD);
Shown after being inoculated with Antagonistic Fungus bacterial strain-penicillium janthinellum by the detection of Disease Resistance-Related Enzymes, in asarum blade
For the activity of PPO, PAL, POD, SOD compared with not being inoculated with Antagonistic Fungus bacterial strain-penicillium janthinellum asarum blade, enzymatic activity is aobvious
It writes and rises, show that Antagonistic Fungus bacterial strain-penicillium janthinellum can induce the raising of Disease Resistance-Related Enzymes in asarum plant body
(as shown in Fig. 5,6,7,8), to resist infecting for disease.
The method of the present invention has biocontrol effect to Pattern of Asarum Leaf Blight by isolating from asarum healthy plant rhizosphere soil
Antagonistic Fungus bacterial strain-penicillium janthinellum, using Pattern of Asarum Leaf Blight bacterium as target bacterium, by tablet face-off method to Antagonistic Fungus bacterial strain
It is screened, the test of living body control efficiency then is carried out to the Antagonistic Fungus bacterial strain filtered out, has obtained Antagonistic Fungus bacterial strain
Preventing preventive effect is 70.78%, and treatment preventive effect is 61.03%, and Antagonistic Fungus bacterial strain is after prevention Pattern of Asarum Leaf Blight occurs and occurs
Control state of an illness extension aspect has preferable effect, and can induce the raising of Disease Resistance-Related Enzymes in asarum body, from
And infecting for disease has been resisted, science data support is provided for the biological control prevention and treatment of Pattern of Asarum Leaf Blight, can be effectively facilitated thin
The development of pungent leaf blight biological control.
The basic principles, main features and advantages of the invention have been shown and described above.The technical staff of the industry should
Understand, the present invention is not limited to the above embodiments, and the above embodiments and description only describe originals of the invention
Reason, without departing from the spirit and scope of the present invention, various changes and improvements may be made to the invention, these changes and improvements
It all fall within the protetion scope of the claimed invention.The claimed scope of the invention is by appended claims and its equivalent circle
It is fixed.
Claims (5)
1. a kind of Antagonistic Fungus bacterial strain for preventing and treating Pattern of Asarum Leaf Blight, it is characterised in that: the Antagonistic Fungus bacterial strain is from healthy asarum
It is isolated in plant rhizosphere soil, it is used for antagonism Pattern of Asarum Leaf Blight pathogen;Classification naming are as follows: microassembly robot
(Penicillium janthinellum);Deposit number: CGMCC No.18101;The preservation time: on June 21st, 2019;It protects
Hide address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3, Institute of Microorganism, Academia Sinica;Depositary institution: the micro- life of China
Object culture presevation administration committee common micro-organisms center.
2. a kind of Antagonistic Fungus bacterial strain for preventing and treating Pattern of Asarum Leaf Blight according to claim 1, it is characterised in that: the antagonism
The filtered fluid of the strain culture of fungal bacterial strain, fermentation liquid or fermentation culture.
3. a kind of Antagonistic Fungus bacterial strain for preventing and treating Pattern of Asarum Leaf Blight according to claim 1, it is characterised in that: the antagonism
Fungal bacterial strain is application of the bacteria agent of active constituent in Pattern of Asarum Leaf Blight prevention and treatment.
4. a kind of Antagonistic Fungus bacterial strain for preventing and treating Pattern of Asarum Leaf Blight according to claim 1, it is characterised in that: the antagonism
Fungal bacterial strain controls the occurrence and development of Pattern of Asarum Leaf Blight, has and prevents and treats effect.
5. a kind of Antagonistic Fungus bacterial strain for preventing and treating Pattern of Asarum Leaf Blight as described in claim 1 is in prevention and treatment pathogen-M. acerina
Application.
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CN113502228A (en) * | 2021-06-22 | 2021-10-15 | 浙江大学 | Penicillium janthinellum strain, biocontrol agent and application |
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