CN105483022B - Application of the sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 in prevention and treatment Alternaria panax - Google Patents
Application of the sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 in prevention and treatment Alternaria panax Download PDFInfo
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Abstract
The invention discloses a kind of sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1, the classification naming of sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 is fusarium solanae (Fusarium solani) TRXY-34-1, the ITS sequence of bacterial strain is as described in SEQ ID NO.1, depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, preservation date: on May 13rd, 2015, deposit number: CGMCC No.10767.The present invention for the first time from the root of medicinal plant sophora tonkinensis Gapnep separation screening to one plant of endogenetic fungus (Fusarium solani) TRXY-34-1, the bacterium has very strong inhibiting effect to Alternaria panax bacterium, brings wide application prospect for the field of biological control of Radix Notoginseng fungal disease.
Description
Technical field
The present invention relates to field of biotechnology, in particular to a kind of sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 is in prevention and treatment Radix Notoginseng
Application in black spot.
Background technique
Modern agriculture is overly dependent upon the use of chemical pesticide to the prevention and treatment of plant disease, and the release of a large amount of chemical pesticides is not
Only there is long-term destruction to ecological environment, also quality of agricultural product is caused to decline, excessive pesticide residues, the drug resistance of pathogen
Property and to problems such as people and animals' nocuousness.It finds safer, effective pest control method to be of great significance, utilize
The method of biology, which carrys out controlling plant diseases, effectively to solve the above problems.
Radix Notoginseng (Panax notoginseng F.H.chen) also known as pseudo-ginseng, invaluable etc., have significant activating microcirculation and removing stasis medicinal,
Detumescence ding-tong effect is a kind of Chinese tradition rare medicinal herbs.Using Radix Notoginseng as Chinese patent drug made of primary raw material such as " Yunnan Baiyao "
" Pien Tze Huang " etc. is wide-spread.In recent years, the demand to Radix Notoginseng raw material is growing.But the nosomycosis in cultivation
Evil has seriously affected Panax notoginseng Growth.Currently, prevention and treatment fungal disease depends on chemical pesticide unduly on notoginseng planting, a large amount of chemistry
The use of pesticide not only affects the quality of Radix Notoginseng, also results in a large amount of pesticide residues of pseudo-ginseng.
Alternaria panax can infect Radix Notoginseng plant and underground root system, aggrieved heavier with stem, leaf, floral axis, be in after root infection
Brown web rot shape, the four seasons can occur, and Radix Notoginseng garden temperature of shed is high, humidity is big, and improper fertilization is likely to add disease sprawling
Weight, general long-term disease incidence is between 5%-20%, up to 60% or more when serious.Its symptom is mostly by Alternaria panax bacterium
Caused by Alternaria panax Whetzel (abbreviation A.panax) harm.
This fungal disease is one of the main reason for causing Radix Notoginseng product quality for many years and yield to decline.In Guangxi three
Seven traditional Genuine producing area, due to low altitude area and the meteorological condition of high temperature and humidity, this disease is often simultaneous, causes
The a large amount of underproduction of Radix Notoginseng are even had no harvest, and are had brought tremendous economic losses to farmer.In order to control this fungal disease, greatly
The chemical pesticide of amount is used, and a large amount of pesticide residues of Radix Notoginseng product are caused, serious to polluted environment, is greatly threatened
People's health.Therefore, development biological control Radix Notoginseng fungal disease is very urgent to the sustainable development of Radix Notoginseng industry and must
It wants.The Antagonistic Fungi for filtering out energy while this disease of antagonism thus is of great significance, and screens and then develops and uses Antagonistic Fungi
It is also effectively to control one of most potential control measure of Radix Notoginseng fungal disease.
The information disclosed in the background technology section is intended only to increase the understanding to general background of the invention, without answering
When being considered as recognizing or imply that the information constitutes the prior art already known to those of ordinary skill in the art in any form.
Summary of the invention
The purpose of the present invention is to provide a kind of sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 answering in prevention and treatment Alternaria panax
With so as to the Alternaria panax occurred during effectively preventing notoginseng planting, to improve the yield and quality of Radix Notoginseng.
To achieve the above object, technical solution provided by the invention is as follows:
The classification naming of a kind of sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1, sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 are Beancurd sheet sickle
Spore (Fusarium solani) TRXY-34-1, the ITS sequence of bacterial strain is as described in SEQ ID NO.1, depositary institution: the micro- life of China
Object culture presevation administration committee common micro-organisms center, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 China
Institute of microbiology, the academy of sciences, preservation date: on May 13rd, 2015, deposit number: CGMCC No.10767.
Preferably, the sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 metabolite the preparation method comprises the following steps:
(1) sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 is inoculated in potato dextrose agar plane ware (PDA is flat
Plate) in, being placed in temperature is to cultivate 20 days at 28 DEG C, and gained culture materials are cut into bulk and are transferred in sterile solid culture medium,
28 DEG C are placed in ferment 60 days；
(2) after fermentation, the methanol soaking fermentation object and ultrasound 40min with 2 times of fermentation material, is then filtered；
(3) step (2) are repeated twice, merges filtrate twice and medicinal extract is concentrated under reduced pressure into, obtains endogenetic fungus TRXY-
The methanol crude extract of 34-1 fermentation material, as sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 metabolite.
Preferably, sterile solid culture medium potato containing 400g described in step (1), the dextrose and 20g sugarcane of 20g
Sugar.
The answering in prevention and treatment Alternaria panax of the metabolite of sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 as obtained by above-mentioned preparation
With.
Compared with prior art, the invention has the following beneficial effects:
The present invention for the first time from the root of medicinal plant sophora tonkinensis Gapnep separation screening to one plant of endogenetic fungus (Fusarium
Solani) TRXY-34-1, the bacterium have very strong inhibiting effect to Alternaria panax bacterium, are that the biology of Radix Notoginseng fungal disease is anti-
Bring wide application prospect in the field of controlling.
Detailed description of the invention
Fig. 1 is the plate opposite culture of sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 and Alternaria panax of the present invention, and wherein F is
Alternaria panax bacterium (Alternaria panax Whetzel), a is blank control, and b is experimental group.
Fig. 2 is sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 strain morphology feature of the present invention, wherein a1 is colonial morphology, and b1 is
Thalli morphology.
Fig. 3 is systematic evolution tree of the sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 bacterial strain based on ITS sequence.
Fig. 4 is the metabolite of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 according to the present invention to Alternaria panax bacterium
The inhibitory effect of mycelia growth；Wherein the 1st, the 5th for blank control be not drug containing PDA plate；2nd, the 3rd, the 4th is sun
Property control powder of carbendazim；6th, the 7th, the 8th is the metabolite of bacterial strain TRXY-34-1, and concentration is respectively 2mg/ml, 4mg/
Ml, 8mg/ml；L is Alternaria panax bacterium Alternaria panax Whetz.
Specific embodiment
It is described in detail With reference to embodiment, it is to be understood that protection scope of the present invention is not by specific
The limitation of embodiment.Experimental method used in following embodiments is conventional method unless otherwise specified.Following implementations
Material, reagent used in example etc., is commercially available unless otherwise specified.Methanol is the commercially available pure methanol of analysis.
2X TagMasterMix is purchased from precious bioengineering Co., Ltd (Takara), and Primer-1, Primer-2 are closed by Hua Da gene
At.
Embodiment 1
Separation, the screening and identification of bacterial strain
One, the separation of bacterial strain
Material to be tested: the wild sophora tonkinensis Gapnep of Guangxi Tiandeng County In Limestone Area is picked up from.
Strains tested: it is provided by plant pathology research institute, Guangxi University.
Culture medium: 1000ml potato dextrose agar (PDA culture medium), PDA culture medium 300g containing potato,
Glucose 20g, agar 20g and chloramphenicol 0.1g, Medium's PH Value are 6.0 ± 0.2.
Surface sterilization: a length of 6-8cm, width is dry to rush for the sophora tonkinensis Gapnep root flowing water flushing 30min of 1-2cm fresh and healthy
Net silt air-dries surface moisture, superclean bench is moved on to, sterile then by clean sophora tonkinensis Gapnep root with rinsed with sterile water 2 times
Under the conditions of, by sophora tonkinensis Gapnep root volumetric concentration be 75% ethyl alcohol impregnate 1min, rinsed with sterile water 2 times, sodium hypochlorite (effective chlorine
1%) 2min is impregnated, sterile washing 3 times, it is spare that sterile blotting paper blots surface.
The separation of bacterial strain purifies: the good sophora tonkinensis Gapnep root of surface sterilization, under aseptic condition, scrapes off epidermis with sterile wood chip, uses
Sterile secateurs cuts off the both ends of sophora tonkinensis Gapnep root, and the sterile pocket knife of remaining part and aseptic nipper separate xylem and bast, and
The tissue block of 0.5cm long is cut into sterile secateurs, it is sterile to be forwarded to the PDA culture medium (containing chloramphenicol) prepared, 28 DEG C of cultures.
Surface sterilization last time rinsing liquid is taken to be coated on PDA plate, as negative control.Observation daily, wait be grown around organizing
Mycelia, the single mycelia of picking, is connected to the PDA culture medium of antibiotic-free.The bacterium colony grown continues if colonial morphology is inconsistent
The single mycelia switching PDA culture medium of picking, until the formalness of the bacterium colony newly grown on plate is consistent.
The endogenetic fungus being separated to is inoculated in PDA plate, it is stand-by after being cultivated 20 days at 28 DEG C.By Alternaria panax bacterium
It is inoculated in PDA plate, it is stand-by after being cultivated 3 days at 28 DEG C.
Two, it screens
Using tablet face-off method to the primary dcreening operation for carrying out thallus bacteriostatic activity for examination sophora tonkinensis Gapnep endogenetic fungus.
Firstly, 6mm bacteria cake is made in endogenetic fungus and Alternaria panax bacterium with sterilization punchers under aseptic condition；Locating
In reason group, endogenetic fungus bacteria cake is transferred to the center of the culture dish containing PDA, then the isometrical 3cm around the bacteria cake of endogenetic fungus
Alternaria panax bacterium bacteria cake is placed at place, and every plant of endogenetic fungus repeats 3 wares；In control group, the culture dish center of PDA does not connect interior life
Fungi bacteria cake places Alternaria panax bacterium bacteria cake at the culture dish center isometrical 3cm of surrounding of PDA, repeats 3 wares.Then 28 DEG C
Lower culture, periodic observation.Radix Notoginseng blackspot is measured in processing group to the long culture dish plate center to PDA of mycelia in control group
The growth radius of germ bacteria cake center to Alternaria panax bacterium between endogenetic fungus bacteria cake center is processing growth radius；It is compareing
In group, the growth radius at measurement Alternaria panax bacterium bacteria cake center to Alternaria panax bacterium between the culture dish center of PDA is pair
According to growth radius.Finally, according to the following formula, calculating bacteriostasis rate:
It compares increment=control and grows radius-bacteria cake radius
It handles increment=processing and grows radius-bacteria cake radius
As a result 4 plants are obtained to all very strong antagonism sophora tonkinensis Gapnep endogenetic fungal bacterial strain of Alternaria panax bacterium inhibiting effect,
In one plant of entitled TRXY-34-1, be 68% to the inhibiting rate of Alternaria panax bacterium.
Three, it identifies
(1) strain morphology feature
Morphological features observation: endophyte fungal bacterial strain TRXY-34-1 to be identified is seeded in PDA culture medium,
28 DEG C of cultures are placed in, observed its cultural characteristic and color change at 5,14 and 20 days respectively.The color characteristic for stablizing maturation is taken to make
Foundation for its cultural characteristic, as identification.Observe and record the color of aerial hyphae, the size of bacterium colony, color, tissue profile,
Surface shape etc. is used as fixed reference feature.
Spore shape observation of characteristics: endophyte fungal bacterial strain TRXY-34-1 to be identified is made into inserted sheet culture, when in PDA
Spore is not produced on culture medium, and mycelia is transferred on the low nutrition culture medium (a quarter concentration PDA) containing sterile sophora tonkinensis Gapnep root
With inducing spore, by observed form result microphotograph, as shown in Figure 2.
(2) bacterial strain ITS sequence and its phylogenetic analysis
The preparation of DNA profiling:
Reagent: (1) lysis buffer: 1% (w/ of Tris-Ac (pH 7.8) 40mM, NaAc 20mM, EDTA 1mM, SDS
v)；
(2) 5M NaCl solution.
DNA is extracted:
Add 600 μ L extracting solutions (Tris-Ac (pH 7.8) 40mM, NaAc 20mM, EDTA 1mM, SDS 1%) to 1.5ml
Centrifuge tube (EP pipe) is put into EP pipe with a small amount of mycelia of stick scraping and grinds, 65 DEG C of water-bath 30min, centrifugal rotational speed 12000r/min, from
Heart 10min；
Centrifugation gained 400 μ L of supernatant is taken, 100 μ L of 5M NaCL, ice bath 10min are added；
Then it is 12000r/min that centrifugal rotational speed is kept at being 4 DEG C in temperature, is centrifuged 10min, takes supernatant；In addition clear liquid
0.6 times of isopropanol and supernatant mix (or 2 times of dehydrated alcohols) ice bath 1h, keep centrifugal rotational speed 12000r/min, centrifugation
10min, the remaining substance of gained dries after discarding supernatant liquid, adds 20 μ L distilled water (ddH2O), 1-2 μ L is taken to do DNA profiling.
PCR amplification ITS sequence:
(1) PCR instrument: ABI 3730-XL DNA sequencer (Applied Biosystems, USA)；
(2) amplimer: ITS1 (5 '-T C C G T A G G T G A A C C T G C G G-3 ') such as SEQ ID
Shown in NO.2 and ITS4 (5 '-T C C T C C G C T T A T T G A T A T G C-3 ') is such as SEQ ID NO.3 institute
Show；
(3) amplification system:
ITS sequence PCR amplification system is as shown in table 1: table 1.
| Reactant | Sample-adding amount |
| 2X TagMasterMix | 25μL |
| Primer-1 | 1μL |
| Primer-2 | 1μL |
| Template DNA | 2μL |
| ddH2O | 21μL |
| React total volume | 50μL |
Note: DNA profiling is made to be above-mentioned in the Template DNA in table 1.
PCR reaction condition is as shown in table 2:
Table 2.
Note: step 2 carries out 30 circulations in table 2.
The electrophoresis detection of pcr amplification product:
The Ago-Gel (the 5 μ l/100ml containing Goldview) that deposition condition is 1%, 1 × TBE electrophoretic buffer, 90V
Electrophoresis 1 hour, PCR product applied sample amount was 3 μ L, point sample after mixing with 1 μ L Loading dye.
In the ultraviolet lower observation of 254nm as a result, using the DL1000DNA Marker of TaKaRa company as nucleic acid standard molecular weight
Object of reference determines expanding fragment length.Amplified production band should be on the position of reference substance 400-700bp.
PCR product purifying and sequencing: it is carried out by Shenzhen Huada Genetic Technology Co., Ltd.
The building of systematic evolution tree:
The ITS sequence of the fungi measured in the ITS sequence surveyed and GenBank gene pool is compared, according to than
Correlated series are downloaded to result.Network analysis is carried out simultaneously by the adjoining algorithm (Neighbor-joining NJ) of MEGA 6.0
Systematic evolution tree is constructed, as shown in Figure 3.
As a result:
(1) strain morphology feature
Colony morphology characteristic: in PDA culture medium, bacterium colony is round, and substrate mycelium canescence is radial, aerial hyphae white,
Flocculence, as shown in a1 in Fig. 2.
Spore shape feature: most of falcate, accidental bar shaped；Macroconidium: (5.00~6.25) μ m
(23.75~40.00) μm；Microconidia: (3.75~5.00) μ m (8.75~13.75) μm, as shown in b1 in Fig. 2.
(2) bacterial strain ITS sequence and its phylogenetic analysis
Using primer I TS1 and ITS4, the segment of a 500-600bp size is amplified from strain gene group DNA, is passed through
It is sequenced and compares sequencing result by BLASTn in GenBank.The result shows that in bacterial strain TRXY-34-1 and Fusarium category
Fungi have a very high base sequence similitude, therefore the reference strain sequence that Fusarium belongs in GenBank is downloaded, for being
Unite developmental analysis, with Thelonectria belong in Thelonectria westlandica (KF569844) and
Thelonectria trachosa (KF569842) is used as outer group.In the systematic evolution tree of building, TRXY-34-1 with
Multiple strains of Fusarium solani and Fusarium oxysporum get together to form the end that holding strength is 99%
Branch.Base similarity system design the result shows that, TRXY-34-1 and Fusarium solani and Fusarium oxysporum alkali
Basic sequence does not have difference, sequence similarity 100%.Comprehensive morphological and molecular biological characteristics, bacterial strain is initially identified as
Fusarium solani。
Embodiment 2
Inhibiting effect of the metabolite of bacterial strain to Alternaria panax bacterium
One, the extraction of the fermented and cultured of bacterium and tunning
(1) sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 is inoculated in potato dextrose agar plane ware (PDA is flat
Plate) in, being placed in temperature is to cultivate 20 days at 28 DEG C, and gained culture materials, which are cut into bulk and are transferred to sterile solid culture medium, (to be contained
400g potato, the dextrose of 20g and 20g sucrose) 2 liters of conical flask in, ferment 60 days under the conditions of being placed in 28 DEG C；
(2) after fermentation, the methanol soaking fermentation object and ultrasound 40min with 2 times of fermentation material, then uses filtered through gauze,
Take filtrate；
(3) step (2) are repeated twice, merges filtrate twice and medicinal extract is concentrated under reduced pressure into, obtains endogenetic fungus TRXY-
The methanol crude extract of 34-1 fermentation material, as sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 metabolite.
Two, inhibiting effect of the sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 metabolite to the mycelia growth of Alternaria panax bacterium
The inhibition grown with the metabolite of mycelia growth method measurement bacterial strain TRXY-34-1 to Alternaria panax bacterium mycelia is living
Property.The metabolite of bacterial strain TRXY-34-1 and powder of carbendazim (positive control) are respectively prepared containing 2mg/mL, 4mg/mL,
The drug containing tablet of 8mg/mL concentration metabolite.Aseptically, 6mm bacteria cake is made in Alternaria panax bacterium with punch,
It is processing, the Alternaria panax bacterium bacterium connect with not drug containing tablet center that Alternaria panax bacterium bacteria cake, which is connected to each drug containing tablet center,
Cake is negative control, and in triplicate, all plates are placed in 28 DEG C of cultures for all processing and control.Drug concentration presses powder of carbendazim
Field information concentration setting.When negative control covers with culture dish, the growth of measurement control bacterium colony and processing bacterium colony is straight respectively
Diameter, and according to the following formula, calculate inhibiting rate:
Negative control increment=negative control growth diameter-bacteria cake diameter
Handle increment=processing growth diameter-bacteria cake diameter
Three, minimal inhibitory concentration of the sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 metabolite to Alternaria panax bacterium
With the metabolite of broth dilution method determination bacterial strain TRXY-34-1 to the minimal inhibitory concentration of Alternaria panax bacterium.
5 days Alternaria panax bacterium pure culture biscuits involvng inoculations will be cultivated in PDA culture medium in the PDB Liquid Culture for containing 0.2% Tween 80 (v/v)
In base, 28 DEG C, 150r/min shaking table culture 7 days, culture is then gone into triangular flask, and pour into the nothing containing 0.2% Tween 80
Bacterium distilled water stirs 30min with magnetism stick, solution is filtered with sterile gauze, obtains spores solution, and will with blood counting chamber
Spore concentration is adjusted to every milliliter 104A spore is spare.The metabolite of bacterial strain is dissolved into the generation of 80mg/mL with 1%DMSO
It thanks to product concentration for the treatment of, 5 sterile test tubes is taken to be sequentially arranged on rack for test tube, number is 1,2,3,4,5 respectively, and every pipe adds respectively
Enter the aseptic double-distilled water that 1ml contains 0.2% Tween 80, number then is added in the bacterial strain methanol crude extract solution of 1ml 80mg/mL
In 1 test tube and successively to carry out twice of serial dilution in each pipe, and by the above-mentioned gained spores solution (10 of 1ml4A/ml) point
It is not added in each pipe, to obtain the concentration for the treatment of of final TRXY-34-1 metabolite: 20mg/ml (1 test tube of number),
(number 5 is tried by 10mg/ml (2 test tube of number), 5mg/ml (3 test tube of number), 2.5mg/ml (4 test tube of number), 1.25mg/ml
Pipe).The 8mg/mL Flusilazole of the 1%DMSO and 1ml of 1ml replace metabolite to execute above-mentioned treatment process and conduct respectively
Negative control and positive control, all processing and control are in triplicate.Finally, 28 DEG C of each pipe, 150r/min shaking table culture 5 days.
The MIC value of the metabolite of TRXY-34-1 is the minimum crude extract concentration for completely inhibiting Alternaria panax bacterium visible growth.
As a result:
The metabolite of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 inhibits the percentage that Alternaria panax bacterium mycelia grows
Rate is as shown in table 3:
Table 3.
Note: positive control powder of carbendazim contains 50% carbendazim in table；* indicates that data pass through one-way analysis of variance in table
LSD relatively after, the metabolite and positive control powder of carbendazim of bacterial strain TRXY-34-1 is under same concentrations, in P0.05Water
Flat upper tool significant difference.
The suppression result that the metabolite of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 grows Alternaria panax bacterium mycelia
Show that the metabolite of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 all has very the mycelia growth of Alternaria panax bacterium
Good inhibitory effect, the metabolite of TRXY-34-1 grows Alternaria panax bacterium A.panax mycelia as can be seen from Table 3
Percent inhibition is 93.13-100%.Compared with positive control powder of carbendazim, the methanol crude extract pair of bacterial strain TRXY-34-1
The inhibitory effect of Alternaria panax bacterium A.panax mycelia growth is equivalent with compareing in 2-4mg/ml, in 8mg/ml, compares
According to good.
The minimal inhibitory concentration such as table that the metabolite of sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 grows Alternaria panax bacterium
Shown in 4:
Table 4.
| Processing | MIC(mg/ml) |
| A.panax | |
| Flusilazole | 0.25 |
| TRXY-34-1 | 1.25 |
Note: positive control Flusilazole active constituent content is 400mg/mL in table.
The minimal inhibitory concentration that the metabolite of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 grows Alternaria panax bacterium
It is stronger that test result shows that the metabolite of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 all has Alternaria panax bacterium
Inhibiting effect, the metabolite of sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 is to Alternaria panax bacterium A.panax as can be seen from Table 4
Minimal inhibitory concentration be 2.5mg/ml, be 10 times of positive control.
As may be known from Table 3 and Table 4, true containing very strong suppression in the metabolite of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1
Bacterium or antifungal ingredient, therefore, in the bionomic control of Alternaria panax bacterium, bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1
With potentiality outstanding.
The aforementioned description to specific exemplary embodiment of the invention is in order to illustrate and illustration purpose.These descriptions
It is not wishing to limit the invention to disclosed precise forms, and it will be apparent that according to the above instruction, can much be changed
And variation.The purpose of selecting and describing the exemplary embodiment is that explaining specific principle of the invention and its actually answering
With so that those skilled in the art can be realized and utilize a variety of different exemplary implementation schemes of the invention and
Various chooses and changes.The scope of the present invention is intended to be limited by claims and its equivalents.
Claims (2)
1. a kind of application of metabolite of sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 in prevention and treatment Alternaria panax, feature exist
In: the classification naming of sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 is fusarium solanae (Fusarium solani) TRXY-34-1, preservation
Number: CGMCC No.10767；
The preparation method of the metabolite of the sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1, operating procedure are as follows:
(1) sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 is inoculated in potato dextrose agar plane ware, is placed in temperature
It is to be cultivated 20 days at 28 DEG C, gained culture materials are cut into bulk and are transferred in sterile solid culture medium, are placed in 28 DEG C of fermentations 60
It；
(2) after fermentation, the methanol soaking fermentation object and ultrasound 40min with 2 times of fermentation material, is then filtered；
(3) step (2) are repeated twice, merges filtrate twice and medicinal extract is concentrated under reduced pressure into, obtains endogenetic fungus TRXY-34-1 hair
The methanol crude extract of ferment object, as sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 metabolite.
2. the metabolite of sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 is in prevention and treatment Alternaria panax according to claim 1
Using, it is characterised in that: sterile solid culture medium potato containing 400g described in step (1), the dextrose and 20g sugarcane of 20g
Sugar.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510979217.6A CN105483022B (en) | 2015-12-23 | 2015-12-23 | Application of the sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 in prevention and treatment Alternaria panax |
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| CN201510979217.6A CN105483022B (en) | 2015-12-23 | 2015-12-23 | Application of the sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 in prevention and treatment Alternaria panax |
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| CN105483022A CN105483022A (en) | 2016-04-13 |
| CN105483022B true CN105483022B (en) | 2019-04-05 |
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| CN105483021B (en) * | 2015-12-23 | 2019-04-05 | 广西大学 | Application of the sophora tonkinensis Gapnep endogenetic fungus TRXY-34-1 in prevention and treatment Radix Notoginseng anthracnose |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105483020A (en) * | 2015-12-23 | 2016-04-13 | 广西大学 | Application of sophora tonkinensis endophytic fungus TRXY-34-1 in prevention and treatment of panax notoginseng root rot |
| CN105483021A (en) * | 2015-12-23 | 2016-04-13 | 广西大学 | Application of sophora tonkinensis endophytic fungus TRXY-34-1 in prevention and treatment of panax notoginseng anthrax |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105483020A (en) * | 2015-12-23 | 2016-04-13 | 广西大学 | Application of sophora tonkinensis endophytic fungus TRXY-34-1 in prevention and treatment of panax notoginseng root rot |
| CN105483021A (en) * | 2015-12-23 | 2016-04-13 | 广西大学 | Application of sophora tonkinensis endophytic fungus TRXY-34-1 in prevention and treatment of panax notoginseng anthrax |
Non-Patent Citations (3)
| Title |
|---|
| 人参黑斑病拮抗细菌分离筛选和鉴定;孙卓等;《吉林农业大学学报》;20140504;第36卷(第3期);全文 * |
| 人参黑斑病生防用内生拮抗菌分离鉴定及发酵浓缩液的性质;贾斌等;《中国森林病虫》;20140531;第33卷(第3期);全文 * |
| 人参黑斑病生防菌的分离鉴定及与杀菌剂混用可能性探讨;贾斌等;《延边大学》;20150115;全文 * |
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| CN105483022A (en) | 2016-04-13 |
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