CN105462852A

CN105462852A - Application of sophora tonkinensis endophytic fungus TRXY-46 in preventing and controlling panax notoginseng anthracnose

Info

Publication number: CN105462852A
Application number: CN201510979380.2A
Authority: CN
Inventors: 李良波; 姚裕群; 黄荣韶; 鲁璇
Original assignee: Guangxi University
Current assignee: Guangxi University
Priority date: 2015-12-23
Filing date: 2015-12-23
Publication date: 2016-04-06
Anticipated expiration: 2035-12-23
Also published as: CN105462852B

Abstract

The invention discloses a sophora tonkinensis endophytic fungus TRXY-46. The sophora tonkinensis endophytic fungus TRXY-46 is classified and named as Rhexocercosporidium sp.TRXY-46, wherein the ITS sequence of the strain is as described by SEQ ID NO.1, the depository authority is the General Microbiological Collection Center of China Committee for Culture Collection of Microorganisms, the depository address is Institute of Microbiology, Building 3, No.1, Beichen West Road, Chaoyang District, Beijing, the depository date is 03 December 2014, and the depository number is CGMCC No.10108. According to the sophora tonkinensis endophytic fungus TRXY-46 provided by the invention, a strain of endophytic fungus Rhexocercosporidium sp.TRXY-46 is separated and screened from the root of a medicinal plant sophora tonkinensis for the first time, the Rhexocercosporidium sp.TRXY-46 has a strong inhibiting effect on panax notoginseng colletotrichum gloeosporioides, and a wide application prospect is created for the biological prevention and control field of panax notoginseng fungal diseases.

Description

The application of sophora tonkinensis Gapnep endogenetic fungus TRXY-46 in control pseudo-ginseng anthrax

Technical field

The present invention relates to biological technical field, the particularly application of a kind of sophora tonkinensis Gapnep endogenetic fungus TRXY-46 in control pseudo-ginseng anthrax.

Background technology

The control of modern agriculture to Plant diseases is overly dependent upon the use of chemical pesticide, discharging of a large amount of chemical pesticide not only has long-term destruction to ecotope, also cause quality of agricultural product to decline, pesticide residue exceed standard, the resistance of pathogenic bacteria and to problems such as people and animals are harmful.Find safer, effective pest control method to be significant, utilizing biological method to carry out controlling plant diseases can effectively solve the problem.

Pseudo-ginseng (PanaxnotoginsengF.H.chen) has another name called pseudo-ginseng, invaluable etc., and having promoting blood circulation and removing blood stasis, subduing swelling and relieving pain effect significantly, is a kind of Chinese tradition rare medicinal herbs.Be that the Chinese patent medicine that main raw material is made is as wide-spread in " Yunnan white powder " and " Pien Tze Huang " etc. with pseudo-ginseng.In recent years, growing to the raw-material demand of pseudo-ginseng.But the fungal disease in cultivation has had a strong impact on Panax notoginseng Growth.At present, on notoginseng planting, control fungal disease depends on chemical pesticide unduly, and the use of a large amount of chemical pesticide not only have impact on the quality of pseudo-ginseng, also result in a large amount of pesticide residue of pseudo-ginseng.

Pseudo-ginseng anthrax is one of Panax notoginseng Growth common disease, and seedling stage, Adult plant all can be fallen ill, and blade their early stage shows as the raw tawny spot of blade, easily bores a hole and break in later stage disease portion.Can form tawny depression spot after petiole and stem infect, after fruit infects, browning look rots, and the cause of disease of this disease is Colletotriehumgloeosporioides (being called for short C.gloeosporioides).

This fungal disease is the one of the main reasons causing pseudo-ginseng quality product and production declining for many years.In traditional Genuine producing area of Guangxi pseudo-ginseng, due to the meteorological conditions of low altitude area and high temperature and humidity, this disease is simultaneous often, and a large amount of underproduction causing pseudo-ginseng are even had no harvest, and brings huge financial loss to plantation family.In order to control this fungal disease, a large amount of chemical pesticides is used, and cause a large amount of pesticide residue of pseudo-ginseng product, serious pollutes environment, has threatened the health of people greatly.Therefore, it is very urgent and necessary for carrying out the Sustainable development of biological control pseudo-ginseng fungal disease to pseudo-ginseng industry.Filter out for this reason and can the Antagonistic Fungi of simultaneously this disease of antagonism be significant, screening and then to develop Antagonistic Fungi be also effectively control one of the most potential prophylactico-therapeutic measures of pseudo-ginseng fungal disease.

The information being disclosed in this background technology part is only intended to increase the understanding to general background of the present invention, and should not be regarded as admitting or imply in any form that this information structure has been prior art that persons skilled in the art are known.

Summary of the invention

The object of the present invention is to provide the application of a kind of sophora tonkinensis Gapnep endogenetic fungus TRXY-46 in control pseudo-ginseng anthrax, thus the pseudo-ginseng anthrax occurred in energy effectively preventing notoginseng planting process, thus improve output and the quality of pseudo-ginseng.

For achieving the above object, technical scheme provided by the invention is as follows:

A kind of sophora tonkinensis Gapnep endogenetic fungus TRXY-46, the Classification And Nomenclature of sophora tonkinensis Gapnep endogenetic fungus TRXY-46 is Rhexocercosporidiumsp.TRXY-46, the ITS sequence of bacterial strain is as described in SEQIDNO.1, depositary institution: China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City Institute of Microorganism, Academia Sinica, preservation date: on December 03rd, 2014, preserving number: CGMCCNo.10108.

Preferably, the preparation method of described sophora tonkinensis Gapnep endogenetic fungus TRXY-46 meta-bolites is:

(1) sophora tonkinensis Gapnep endogenetic fungus TRXY-46 is inoculated in potato dextrose agar plane ware (PDA is dull and stereotyped), being placed in temperature is cultivate 20 days at 28 DEG C, gained cultivated material is cut into bulk and transfers in sterile solid substratum, is placed in 28 DEG C of fermentations 60 days;

(2) after having fermented, with the methyl alcohol soaking fermentation thing of fermented product 2 times and ultrasonic 40min, then filter;

(3) repeating step (2) twice, merges twice filtrate and carries out concentrating under reduced pressure and become medicinal extract, obtain the methanol crude extract of endogenetic fungus TRXY-46 fermented product, be sophora tonkinensis Gapnep endogenetic fungus TRXY-46 meta-bolites.

Preferably, the sterile solid substratum described in step (1) contains 400g potato, the dextrose of 20g and 20g sucrose.

The application of meta-bolites in control pseudo-ginseng anthrax of gained sophora tonkinensis Gapnep endogenetic fungus TRXY-46 is prepared as above-mentioned.

Compared with prior art, the present invention has following beneficial effect:

The present invention first from the root of medicinal plant sophora tonkinensis Gapnep separation screening to strain endogenetic fungus (Rhexocercosporidiumsp.) TRXY-46, this bacterium has very strong restraining effect to pseudo-ginseng anthrax bacteria, and the field of biological control for pseudo-ginseng fungal disease brings wide application prospect.

Accompanying drawing explanation

Fig. 1 is the dull and stereotyped opposite culture of sophora tonkinensis Gapnep endogenetic fungus TRXY-46 of the present invention and pseudo-ginseng anthrax, and wherein F is pseudo-ginseng anthrax bacteria (Colletotriehumgloeosporioides), a is blank, and b is experimental group.

Fig. 2 is sophora tonkinensis Gapnep endogenetic fungus TRXY-46 strain morphology feature of the present invention.

Fig. 3 is the systematic evolution tree of sophora tonkinensis Gapnep endogenetic fungus TRXY-46 bacterial strain based on ITS sequence.

Fig. 4 is to the inhibition of the mycelial growth of pseudo-ginseng anthrax bacteria according to the meta-bolites of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-46 of the present invention; Wherein the 1st, the 5th is that namely the PDA of pastille is not dull and stereotyped for blank; 2nd, the 3rd, the 4th is positive control powder of carbendazim; 6th, the 7th, the 8th is the meta-bolites of bacterial strain, and concentration is respectively 2mg/ml, 4mg/ml, 8mg/ml; T is pseudo-ginseng anthrax bacteria Colletotriehumgloeosporioides.

Embodiment

Be described in detail below in conjunction with embodiment, but be to be understood that protection scope of the present invention not by the restriction of embodiment.The experimental technique used in following embodiment if no special instructions, is ordinary method.The material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.Methyl alcohol is commercial analytical pure methyl alcohol.2XTagMasterMix is purchased from precious biotechnology company limited (Takara), and Primer-1, Primer-2 are by Hua Da gene chemical synthesis.

Embodiment 1

The separation of bacterial strain, screening and qualification

One, the separation of bacterial strain

For examination material: the wild sophora tonkinensis Gapnep picking up from In Limestone Area, Tiandeng County, Guangxi.

Strains tested: provided by plant pathology institute of Guangxi University.

Substratum: 1000ml potato dextrose agar (PDA substratum), PDA substratum is containing potato 300g, glucose 20g, agar 20g and paraxin 0.1g, and Medium's PH Value is 6.0 ± 0.2.

Surface sterilization: by long for 6-8cm, the wide sophora tonkinensis Gapnep root running water 30min for 1-2cm fresh and healthy are to rinse silt, then the sophora tonkinensis Gapnep root rinsed with sterile water 2 times will cleaned, air-dry surface-moisture, moving on to Bechtop, aseptically, is 75% alcohol immersion 1min by sophora tonkinensis Gapnep root volumetric concentration, rinsed with sterile water 2 times, clorox (available chlorine 1%) soaks 2min, aseptic washing 3 times, and it is for subsequent use that aseptic thieving paper blots surface.

The separation of bacterial strain, purifying: the sophora tonkinensis Gapnep root that surface sterilization is good, under aseptic condition, epidermis is scraped off with aseptic wood chip, the two ends of sophora tonkinensis Gapnep root are cut off with aseptic secateurs, remaining part aseptic pocket knife and aseptic nipper separate xylem and phloem, and be cut into the long tissue block of 0.5cm with aseptic secateurs, be asepticly forwarded to the PDA substratum (containing paraxin) prepared, 28 DEG C of cultivations.Getting the last rinsing liquid of surface sterilization is coated on PDA flat board, as negative control.Every day is observed, and waits to organize to grow mycelia, the single mycelia of picking around, is connected to the PDA substratum of antibiotic-free.The bacterium colony grown, if colonial morphology is inconsistent, continues picking single mycelia switching PDA substratum, until the formalness of the bacterium colony that flat board newly grows is consistent.

The endogenetic fungus be separated to is inoculated in PDA flat board, cultivates after 20 days stand-by at 28 DEG C.Pseudo-ginseng anthrax bacteria is inoculated in PDA flat board, cultivates after 3 days stand-by at 28 DEG C.

Two, screen

Adopt dull and stereotyped face-off method to the primary dcreening operation carrying out thalline bacteriostatic activity for examination sophora tonkinensis Gapnep endogenetic fungus.

First, under aseptic condition, with sterilizing punch tool, endogenetic fungus and pseudo-ginseng anthrax bacteria are made 6mm bacterium cake; In treatment group, endogenetic fungus bacterium cake is transferred to the culture dish central authorities containing PDA, then around the bacterium cake of endogenetic fungus, pseudo-ginseng anthrax bacteria bacterium cake is placed at isometrical 3cm place, and every strain endogenetic fungus repeats 3 wares; In control group, the culture dish central authorities of PDA do not connect endogenetic fungus bacterium cake, and around the culture dish central authorities of PDA, pseudo-ginseng anthrax bacteria bacterium cake is placed at isometrical 3cm place, repeats 3 wares.Then cultivate at 28 DEG C, periodic observation.Treat that in control group, mycelia grows to the dull and stereotyped central authorities of culture dish of PDA, in treatment group, measure the growth radius of pseudo-ginseng anthrax bacteria between pseudo-ginseng anthrax bacteria Jun Bing center to endogenetic fungus Jun Bing center for process growth radius; In control group, measure pseudo-ginseng anthrax bacteria Jun Bing center to PDA culture dish center between pseudo-ginseng anthrax bacteria growth radius for contrast grow radius.Finally, according to following formula, calculate bacteriostasis rate:

Contrast increment=contrast growth radius-bacterium cake radius

Process increment=process growth radius-bacterium cake radius

Result obtains the 4 strains antagonism sophora tonkinensis Gapnep endogenetic fungal bacterial strain all very strong to pseudo-ginseng anthrax bacteria restraining effect altogether, and wherein a strain name is called TRXY-46, is 60% to the inhibiting rate of pseudo-ginseng anthrax bacteria.

Three, identify

(1) strain morphology feature

Morphological features is observed: be seeded on PDA substratum by endophyte fungal bacterial strain TRXY-46 to be identified, be placed in 28 DEG C of cultivations, observe its cultural characteristic and colour-change respectively at 5,14 and 20 days.Get stable ripe color characteristic as its cultural characteristic, as the foundation of qualification.The color of observed and recorded aerial hyphae, the size, color, tissue profile, surface shape etc. of bacterium colony are as reference feature.

Spore shape observation of characteristics: endophyte fungal bacterial strain TRXY-46 to be identified is made inserted sheet and cultivate, when not producing spore on PDA substratum, mycelia is transferred on the low nutrition substratum (1/4th concentration PDA) containing aseptic sophora tonkinensis Gapnep root with inducing spore, by the photomicrography of viewed form result, as shown in Figure 2.

(2) bacterial strain ITS sequence and phylogenetic analysis thereof

The preparation of DNA profiling:

Reagent: (1) lysis buffer: Tris-Ac (pH7.8) 40mM, NaAc20mM, EDTA1mM, SDS1% (w/v);

(2) 5MNaCl solution.

DNA extraction:

Add extracting solution (Tris-Ac (pH7.8) 40mM of 600 μ L65 DEG C preheatings, NaAc20mM, EDTA1mM, SDS1%) to 1.5ml centrifuge tube (EP pipe), scrape with rod the mycelia that takes a morsel and put into the grinding of EP pipe, 65 DEG C of water-bath 30min, centrifugal rotational speed 12000r/min, centrifugal 10min;

Get centrifugal gained supernatant liquor 400 μ L, add 5MNaCL100 μ L, ice bath 10min;

Then at temperature is 4 DEG C, keep centrifugal rotational speed to be 12000r/min, centrifugal 10min, gets supernatant liquor; Mix (or 2 times of dehydrated alcohols) ice bath 1h with supernatant liquor 0.6 times of Virahol and supernatant liquor, keep centrifugal rotational speed 12000r/min, centrifugal 10min, after abandoning supernatant, the remaining material of gained dries, and adds 20 μ L distilled water (ddH ₂o), get 1-2 μ L and do DNA profiling.

Pcr amplification ITS sequence:

(1) PCR instrument: ABI3730-XLDNA sequenator (AppliedBiosystems, USA)

(2) amplimer: ITS1 (5 '-TCCGTAGGTGAACCTGCGG-3 ') is as shown in SEQIDNO.2, and ITS4 (5 '-TCCTCCGCTTATTGATATGC-3 ') is as shown in SEQIDNO.3;

(3) amplification system:

ITS sequence PCR amplification system is as shown in table 1: table 1.

Reactant	Application of sample amount
		2X TagMasterMix	25μL
Primer-1	1μL
		Primer-2	1μL
Template DNA	2μL
		ddH ₂O	21μL
Reaction cumulative volume	50μL

Note: the TemplateDNA in table 1 above-mentionedly makes DNA profiling.

PCR reaction conditions is as shown in table 2:

Table 2.

Note: in table 2, step 2 carries out 30 circulations.

The electrophoresis detection of pcr amplification product:

Deposition condition is the sepharose (containing Goldview5 μ l/100ml) of 1%, 1 × TBE electrophoretic buffer, 90V electrophoresis 1 hour, and PCR primer applied sample amount is 3 μ L, point sample after mixing with 1 μ LLoadingdye.Observations under 254nm ultraviolet, with the DL1000DNAMarker of TaKaRa company for nucleic acid standard molecular weight object of reference, determines expanding fragment length.Amplified production band should on the position of standard substance 400-700bp.

PCR primer purifying and order-checking: undertaken by Shenzhen Huada Genetic Technology Co., Ltd.

The structure of systematic evolution tree:

The ITS sequence of the fungi measured in surveyed ITS sequence and GenBank gene pool is compared, downloads correlated series according to comparison result.Systems analysis is carried out and constructing system evolutionary tree, as shown in Figure 3 by the adjacent algorithm (Neighbor-joiningNJ) of MEGA6.0.

Result:

(1) strain morphology feature

Bacterium colony size: better than commercial PDA substratum grows on artificial PDA substratum, on artificial PDA, the growth of bacterium colony limitation, poor growth, cultivates one month, bacterium colony fusiform, diameter 16mm, height 3mm;

Colony colour: it is green that bacterium colony is just flushing, and reverse side is pink, and edge is greyish white, and substratum is slightly red;

The tissue profile of bacterium colony: mycelia forms fine hair shape closely, and colony edge is wavy;

The surface shape of bacterium colony: mycelia closely, there is water droplet on bacterium colony surface;

Mycelia: mainly aerial hyphae, without spore, old mycelia is red, and new mycelia is grayish green.

Spore shape feature: through induction, without spore.

(2) bacterial strain ITS sequence and phylogenetic analysis thereof

Utilize primer I TS1 and ITS4, the fragment of a 500-600bp size is amplified from strain gene group DNA, through order-checking and by sequencing result by BLASTn comparison in GenBank, download relevant reference strain sequence according to comparison result, utilize the adjacent algorithm (Neighbor-joiningNJ) of MEGA6 to carry out systems analysis and constructing system evolutionary tree.Result shows, bacterial strain TRXY-46 and Rhexocercosporidiumsp.Dzf14 (EU543257) gets together and forms holding strength is the end branch of 93%.Base similarity system design result shows, TRXY-46 and Rhexocercosporidiumsp.Dzf14 (EU543257) differs 3 bases, and sequence similarity is 99.3%.Comprehensive morphological and molecular biological characteristics, be initially identified as Rhexocercosporidiumsp. by bacterial strain.

Embodiment 2

The meta-bolites of bacterial strain is to the restraining effect of pseudo-ginseng anthrax bacteria

One, the fermentation culture of bacterium and the extraction of tunning

(1) sophora tonkinensis Gapnep endogenetic fungus TRXY-46 is inoculated in potato dextrose agar plane ware (PDA is dull and stereotyped), being placed in temperature is cultivate 20 days at 28 DEG C, gained cultivated material is cut into bulk and transfers in the Erlenmeyer flask of 2 liters of sterile solid substratum (dextrose and 20g sucrose containing 400g potato, 20g), is placed in 28 DEG C of condition bottom fermentations 60 days;

(2) after having fermented, with the methyl alcohol soaking fermentation thing of fermented product 2 times and ultrasonic 40min, then use filtered through gauze, get filtrate;

Two, sophora tonkinensis Gapnep endogenetic fungus TRXY-46 meta-bolites is to the restraining effect of the mycelial growth of pseudo-ginseng anthrax bacteria

The meta-bolites of bacterial strain TRXY-46 is measured to the inhibit activities of pseudo-ginseng anthrax bacteria mycelial growth by mycelial growth method.The pastille meta-bolites of bacterial strain TRXY-46 and powder of carbendazim (positive control) made respectively containing 2mg/mL, 4mg/mL, 8mg/mL concentration meta-bolites is dull and stereotyped.Aseptically, with punch tool, pseudo-ginseng anthrax bacteria is made 6mm bacterium cake, it is process that pseudo-ginseng anthrax bacteria bacterium cake is received the dull and stereotyped central authorities of each pastille, the pseudo-ginseng anthrax bacteria bacterium cake connect with the dull and stereotyped central authorities of not pastille is for negative control, in triplicate, all flat boards are placed in 28 DEG C of cultivations for all process and contrast.The Field information concentration that drug level presses powder of carbendazim is arranged.When negative control covers with culture dish, measure the growth diameter of contrast bacterium colony and process bacterium colony respectively, and according to following formula, calculate inhibiting rate:

Negative control increment=negative control growth diameter-bacterium cake diameter

Process increment=process growth diameter-bacterium cake diameter

Three, sophora tonkinensis Gapnep endogenetic fungus TRXY-46 meta-bolites is to the minimal inhibitory concentration of pseudo-ginseng anthrax bacteria

With the meta-bolites of broth dilution method determination bacterial strain TRXY-46 to the minimal inhibitory concentration of pseudo-ginseng anthrax bacteria.The pseudo-ginseng anthrax bacteria pure culture biscuits involvng inoculation of 5 days will be cultivated in the PDB liquid nutrient medium containing 0.2% tween 80 (v/v) in PDA substratum, 28 DEG C, 150r/min shaking table cultivates 7 days, then culture is forwarded to triangular flask, and the aseptic double-distilled water poured into containing 0.2% tween 80, stir 30min with magnetism stick, solution sterile gauze is filtered, obtain spores solution, and with blood counting chamber, spore concentration is adjusted to every milliliter 10 ⁴individual spore is for subsequent use.The meta-bolites 1%DMSO of bacterial strain TRXY-46 is dissolved into the meta-bolites concentration for the treatment of of 80mg/mL, getting 5 sterile test tube is sequentially arranged on test-tube stand, be numbered 1,2,3,4,5 respectively, often pipe adds the aseptic double-distilled water of 1ml containing 0.2% tween 80 respectively, then the bacterial strain methanol crude extract solution of 1ml80mg/mL is added in the test tube being numbered 1 and also in each pipe, carry out twice serial dilution successively, and by above-mentioned for 1ml gained spores solution (10 ⁴individual/ml) add in each pipe respectively, thus obtain the meta-bolites concentration for the treatment of of final TRXY-46: 20mg/ml (numbering 1 test tube), 10mg/ml (numbering 2 test tube), 5mg/ml (numbering 3 test tube), 2.5mg/ml (numbering 4 test tube), 1.25mg/ml (numbering 5 test tube).The 8mg/mL Flusilazole of 1%DMSO and 1ml of 1ml replaces meta-bolites to perform above-mentioned treating processes respectively and as negative control and positive control, all process and contrast are in triplicate.Finally, each pipe 28 DEG C, 150r/min shaking table cultivates 5 days.The MIC value of the meta-bolites of TRXY-46 is suppress the minimum crude extract concentration of pseudo-ginseng anthrax bacteria visible growth completely.

Result:

The percent inhibition of meta-bolites to pseudo-ginseng anthrax bacteria mycelial growth of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-46 is as shown in table 3:

Table 3.

Note: show positives contrast powder of carbendazim containing 50% derosal; After in table, * represents that LSD that data pass through one-way analysis of variance is relatively, the meta-bolites of bacterial strain TRXY-46 and positive control powder of carbendazim under same concentrations, at P _0.05tool significant difference in level.

The suppression result of meta-bolites to pseudo-ginseng anthrax bacteria mycelial growth of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-46 shows, the meta-bolites of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-46 all has extraordinary inhibition to the mycelial growth of pseudo-ginseng anthrax bacteria, the percent inhibition of meta-bolites to pseudo-ginseng anthrax bacteria C.gloeosporioides mycelial growth of TRXY-46 is 100.00% as can be seen from Table 3, compare with positive control powder of carbendazim, the meta-bolites of bacterial strain TRXY-46 to the inhibition of pseudo-ginseng anthrax bacteria C.gloeosporioides mycelial growth with contrast equivalent.

The minimal inhibitory concentration that sophora tonkinensis Gapnep endogenetic fungus TRXY-46 meta-bolites grows pseudo-ginseng anthrax bacteria is as shown in table 4:

Table 4.

Process	MIC(mg/ml)
			C.gloeosporioides
Flusilazole	0.25
		TRXY-46	1.25

Note: showing positives contrast Flusilazole active constituent content is 400mg/mL.

The meta-bolites of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-46 shows the minimal inhibitory concentration test result that pseudo-ginseng anthrax bacteria grows, the meta-bolites of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-46 all has stronger restraining effect to pseudo-ginseng anthrax bacteria, the minimal inhibitory concentration of meta-bolites to pseudo-ginseng anthrax bacteria C.gloeosporioides of sophora tonkinensis Gapnep endogenetic fungus TRXY-46 is 1.25mg/ml as can be seen from Table 4, is 5 times of positive control.

As may be known from Table 3 and Table 4, contain the composition of very strong antifungal or fungicidal in the meta-bolites of bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-46, therefore, in the bionomic control of pseudo-ginseng anthrax bacteria, bacterial strain sophora tonkinensis Gapnep endogenetic fungus TRXY-46 has outstanding potentiality.

The aforementioned description to concrete exemplary of the present invention is to illustrate and the object of illustration.These descriptions not want the present invention to be defined as disclosed precise forms, and obviously, according to above-mentioned instruction, can much change and change.The object selected exemplary embodiment and describe is to explain certain principles of the present invention and practical application thereof, thus those skilled in the art can be realized and utilize various different exemplary of the present invention and various different selection and change.Scope of the present invention is intended to limited by claims and equivalents thereof.

Claims

1. a sophora tonkinensis Gapnep endogenetic fungus TRXY-46, is characterized in that: the Classification And Nomenclature of sophora tonkinensis Gapnep endogenetic fungus TRXY-46 is Rhexocercosporidiumsp.TRXY-46, preserving number: CGMCCNo.10108.

2. a preparation method for the meta-bolites of sophora tonkinensis Gapnep endogenetic fungus TRXY-46 as claimed in claim 1, it is characterized in that, operation steps is as follows:

(1) be inoculated in potato dextrose agar plane ware by sophora tonkinensis Gapnep endogenetic fungus TRXY-46, being placed in temperature is cultivate 20 days at 28 DEG C, and gained cultivated material is cut into bulk and transfers in sterile solid substratum, is placed in 28 DEG C of fermentations 60 days;

3. the preparation method of meta-bolites according to claim 2, is characterized in that: the sterile solid substratum described in step (1) containing 400g potato, the dextrose of 20g and 20g sucrose.

4. the application of meta-bolites in control pseudo-ginseng anthrax preparing gained sophora tonkinensis Gapnep endogenetic fungus TRXY-46 as claim 2.