CN105462851B - Application of the sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 in prevention and treatment notoginseng root rot - Google Patents

Application of the sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 in prevention and treatment notoginseng root rot Download PDF

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CN105462851B
CN105462851B CN201510979246.2A CN201510979246A CN105462851B CN 105462851 B CN105462851 B CN 105462851B CN 201510979246 A CN201510979246 A CN 201510979246A CN 105462851 B CN105462851 B CN 105462851B
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endogenetic fungus
sophora tonkinensis
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韦继光
姚裕群
黄荣韶
李良波
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Guangxi University
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Abstract

The invention discloses a kind of sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2, the classification naming of sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 is Rhexocercosporidium sp.TRXY-59-2, 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 December 03rd, 2014, deposit number: CGMCC No.10109.The present invention for the first time from the root of medicinal plant sophora tonkinensis Gapnep separation screening to one plant of endogenetic fungus (Rhexocercosporidium sp.) TRXY-59-2, the bacterium has very strong inhibiting effect to notoginseng root rot bacterium, brings wide application prospect for the field of biological control of Radix Notoginseng fungal disease.

Description

Application of the sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 in prevention and treatment notoginseng root rot
Technical field
The present invention relates to field of biotechnology, in particular to a kind of sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 is in prevention and treatment Radix Notoginseng Application in root rot.
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.
Notoginseng root rot is one of Radix Notoginseng Major Diseases, is commonly called as " green smelly " in producing region, cardinal symptom be seedling stage bud-rot and The water stain shape lesion of brown of its reed head and bastem junction is until spread to entire stem base portion.Just there is generation in the disease seeding stage, 4, disease in May is stagnated, and the 6-9 month enters rainy season, into onset peak period.There are many types for root rot, it was reported that its symptom Mostly caused by notoginseng root rot bacterium (Fusarium solani) (abbreviation F.solani).
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-59-2 answering in prevention and treatment notoginseng root rot With so as to the notoginseng root rot 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-59-2, sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 is Rhexocercosporidium sp.TRXY-59-2, the ITS sequence of bacterial strain is as described in SEQ ID NO.1, depositary institution: China Microbiological Culture Collection administration committee common micro-organisms center, preservation address: Yard 1, BeiChen xi Road, Chaoyang District, Beijing City 3 Institute of Microorganism, Academia Sinica, preservation date: on December 03rd, 2014, deposit number: CGMCC No.10109.Preferably Be, the sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 metabolite the preparation method comprises the following steps:
(1) sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 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 59-2 fermentation material, as sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 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 notoginseng root rot of the metabolite of sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 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 (Rhexocercosporidium sp.) TRXY-59-2, the bacterium have very strong inhibiting effect to notoginseng root rot bacterium, are three The field of biological control of seven fungal diseases brings wide application prospect.
Detailed description of the invention
Fig. 1 is the plate opposite culture of sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 and notoginseng root rot of the present invention, and wherein F is Notoginseng root rot bacterium (Fusarium solani), a are blank control, and b is experimental group.
Fig. 2 is sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 strain morphology feature of the present invention.
Fig. 3 is systematic evolution tree of the sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 bacterial strain based on ITS sequence.
Fig. 4 is the metabolite of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 according to the present invention to notoginseng root rot 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, and concentration is respectively 2mg/ml, 4mg/ml, 8mg/ ml；F is notoginseng root rot bacterium Fusarium solani.
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. Last time rinsing liquid is taken to be coated on PDA plate, as negative control.Observation daily, wait grow mycelia, picking around organizing Single mycelia is connected to the PDA culture medium of antibiotic-free.The bacterium colony grown continues the single bacterium of picking if formalness is inconsistent Silk switching PDA culture medium, 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 notoginseng root rot 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 notoginseng root rot 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 Notoginseng root rot 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 notoginseng root rot 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 root-rot 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 notoginseng root rot bacterium between endogenetic fungus bacteria cake center is processing growth radius；It is compareing In group, the growth radius at measurement notoginseng root rot bacterium bacteria cake center to notoginseng root rot 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 notoginseng root rot bacterium inhibiting effect, In one plant of entitled TRXY-59-2, be 70% to the inhibiting rate of notoginseng root rot bacterium.
Three, it identifies
(1) strain morphology feature
Morphological features observation: endophyte fungal bacterial strain TRXY-59-2 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-59-2 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:
Extracting solution (Tris-Ac (pH 7.8) 40mM, NaAc 20mM, the EDTA 1mM, SDS for adding 600 μ L65 DEG C to preheat 1%) 1.5ml centrifuge tube (EP pipe) is arrived, scrapes a small amount of mycelia with stick and is put into the grinding of EP pipe, 65 DEG C of water-bath 30min, centrifugal rotational speed 12000r/min is centrifuged 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 sight of 254nm It examines as a result, determining that amplified fragments are long using the DL1000 DNA Marker of TaKaRa company as nucleic acid standard molecular weight object of reference Degree.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 and structure are carried out by the adjoining algorithm (Neighbor-joining NJ) of MEGA6.0 Systematic evolution tree is built, as shown in Figure 3.
As a result:
(1) strain morphology feature
Bacterium colony size: it is better than being grown in the PDA culture medium of commercialization in artificial PDA culture medium, on artificial PDA, bacterium Limitation growth is fallen, slow growth is cultivated one month, bacterium colony fusiform, diameter 20mm, height 5mm；
Colony colour: bacterium colony front is sea spray, and reverse side center is yellowish green, and edge is dark green, and culture medium is slightly red；
The tissue profile of bacterium colony: new mycelia forms close carpet-like, and old mycelia villiform, colony edge is wavy,；
The surface shape of bacterium colony: new mycelia is close, and old mycelia villiform, there is water droplet on bacterium colony surface；
Mycelia: matrix mycelia is blackish green, and aerial hyphae is sea spray, no spore.Old mycelia is greyish white, and new mycelia is grayish green.
Spore shape feature: through inducing, no spore.
(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, relevant reference strain sequence, benefit are downloaded according to comparison result Network analysis is carried out with the adjoining algorithm (Neighbor-joining NJ) of MEGA6 and constructs systematic evolution tree.The result shows that Bacterial strain TRXY-59-2 and Rhexocercosporidium sp.Dzf14 (EU543257), which gets together, to be formed holding strength and is 99% end branch.Base similarity system design the result shows that, TRXY-59-2 and Rhexocercosporidium sp.Dzf14 (EU543257) there is no difference, sequence similarity 100%.Comprehensive morphological and molecular biological characteristics tentatively reflect bacterial strain It is set to Rhexocercosporidium sp..
Embodiment 2
Inhibiting effect of the metabolite of bacterial strain to notoginseng root rot bacterium
One, the extraction of the fermented and cultured of bacterium and tunning
(1) sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 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 59-2 fermentation material, as sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 metabolite.
Two, inhibiting effect of the sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 metabolite to the mycelia growth of notoginseng root rot bacterium
The inhibition grown with the metabolite of mycelia growth method measurement bacterial strain TRXY-59-2 to notoginseng root rot bacterium mycelia is living Property.The metabolite of bacterial strain TRXY-59-2 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 notoginseng root rot bacterium with punch, It is processing, the notoginseng root rot bacterium bacterium connect with not drug containing tablet center that notoginseng root rot 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-59-2 metabolite to notoginseng root rot bacterium
With the metabolite of broth dilution method determination bacterial strain TRXY-59-2 to the minimal inhibitory concentration of notoginseng root rot bacterium. 5 days notoginseng root rot 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 TRXY-59-2 is dissolved into 1%DMSO The metabolite concentration for the treatment of of 80mg/mL takes 5 sterile test tubes to be sequentially arranged on rack for test tube, respectively number be 1,2,3,4, 5, every pipe is separately added into the aseptic double-distilled water that 1ml contains 0.2% Tween 80, then by the bacterial strain methanol crude extract of 1ml 80mg/mL Solution is added in the test tube that number is 1 and successively carries out twice of serial dilution in each pipe, and by the above-mentioned gained spores solution of 1ml (104A/ml) it is separately added into each pipe, to obtain final metabolite concentration for the treatment of: 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-59-2 is the minimum crude extract concentration for completely inhibiting notoginseng root rot bacterium visible growth.
As a result:
The metabolite of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 inhibits the percentage that notoginseng root rot 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-59-2 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-59-2 grows notoginseng root rot bacterium mycelia Show that the metabolite of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 all has very the mycelia growth of notoginseng root rot bacterium Good inhibitory effect, the metabolite of TRXY-59-2 grows notoginseng root rot bacterium F.solani mycelia as can be seen from Table 3 Percent inhibition is 93.93-94.06%, and compared with positive control powder of carbendazim, the metabolite of bacterial strain TRXY-59-2 is right The inhibitory effect of notoginseng root rot bacterium F.solani mycelia growth slightly below compares.
The minimal inhibitory concentration that bacterial strain TRXY-59-2 metabolite grows notoginseng root rot bacterium is as shown in table 4:
Table 4.
Processing MIC(mg/ml)
F.solani
Flusilazole 0.5
TRXY-59-2 10
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-59-2 grows notoginseng root rot bacterium It is stronger that test result shows that the metabolite of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 all has notoginseng root rot bacterium Inhibiting effect, the metabolite of sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 is to notoginseng root rot bacterium F.solani as can be seen from Table 4 Minimal inhibitory concentration be 10mg/ml, be 20 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-59-2 Bacterium or antifungal ingredient, therefore, in the bionomic control of notoginseng root rot bacterium, bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 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 (1)

1. a kind of application of metabolite of sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 in prevention and treatment notoginseng root rot, feature exist In: the classification naming of the sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 isRhexocercosporidium sp. TRXY-59-2, Deposit number: CGMCC No. 10109；
The preparation method of the metabolite of the sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2, operating procedure are as follows:
(1) sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 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；Wherein, the sterile solid culture medium potato containing 400g, the dextrose and 20g sucrose of 20g；
(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-59-2 hair The methanol crude extract of ferment object, as sophora tonkinensis Gapnep endogenetic fungus TRXY-59-2 metabolite.
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