CN102978128A - Antagonistic bacterium cooperating with AMF for disease resisting and growth promoting and application thereof - Google Patents
Antagonistic bacterium cooperating with AMF for disease resisting and growth promoting and application thereof Download PDFInfo
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Abstract
The invention discloses an antagonistic bacterium cooperating with AMF for disease resisting and growth promoting and application thereof. The antagonistic bacterium is bacillus amyloliquefaciens with an accession number of CGMCC No. 6491. According to the invention, bacterial strains with strong bacteriostatic activity are obtained by carrying out screening of the antagonistic bacterium on soil samples collected from a farmland cultivated for a plurality of years in a suburban area of Fukang, Xinjiang; then further screening and domestication are carried out, and the taxonomic status of obtained stains are preliminarily determined through determination of morphological characteristics and the 16 S rRNA sequence of the strains and phylogenetic analysis of the strains; moreover, in-depth research is carried out on characteristics like a bacteriostasis rate and stability of the fermentation broth of the bacterium. The antagonistic bacterium offers a new option for selection of antagonistic bacteria capable of controlling soil-borne diseases of vegetable; antagonistic action of the antagonistic bacterium exerts a prominent technical effect on treating tomato wilt, fusarium wilt of cucumber, phytophthora capsici leon and pepper anthracnose; and the antagonistic bacterium has wide application values.
Description
Technical field
The present invention relates to agriculture technical field of microbe application, specifically, the present invention relates to Antagonistic Fungi and the applied technical field in the control soil-borne diseases of vegetable thereof a kind of and the collaborative disease-resistant short life of AMF.
Background technology
In Xinjiang because geography and the weather condition of its uniqueness, industrialized agriculture becomes one of Main Patterns of growing vegetables, and along with the attention to vegetable basket project of country and autonomous region, the tempo of industrialized agriculture and scale improve year by year and enlarge, but because the industrialized agriculture disease and pest is easily sent out, reality rambunctious, therefore also have higher requirement for the prevention and control of disease and pest, especially the vegetable disease in the industrialized agriculture is particularly serious, and soil-borne disease is one of major objective of prevention and control; Soil-borne disease refers to live in the germ in the residual body of diseased plant in pathogenic agent in the soil or the soil, the disease that causes from crop root or stem's infringement; The industrialized agriculture soil-borne diseases of vegetable mainly contains vertical withered, the samping off in root knot nematode disease, cucurbits fusarium wilt, verticillium, seedling stage etc.; The industrialized agriculture vegetable growing, owing to limited by geographical conditions, the crop rotation difficulty, the continuous cropping cycle is long, adds the conditions favourings such as temperature, humidity in generation, the propagation of various diseases, and the soil-borne disease difficulty of prevention and cure is larger, in recent years, its sickness rate is high, hazardness is large, has become the major obstacle factor that affects industrialized agriculture vegetables productivity effect, is seriously restricting the development of vegetables industry; AM fungi bush mycorrhizal fungi (
Arbuscular Mycorrhizal Fungi, be called for short AMF, the AM fungi is once called as the VA bacterium) be prevalent in the soil all over the world, its host comprises bryophyte, pteridophyte, gymnosperm and angiosperm; According to statistics, on the earth in 90% vascular plant and the agroecosystem most of crops can both form arbuscular mycorrhiza; Bush mycorrhizal fungi and plant symbiosis can increase the absorption area of plant root, and increase particularly to the absorption of phosphorus element, therefore can promote the absorption of plant shoot and root to mineral nutrition, can also improve the resistance of plant simultaneously; For the growth of crop, the control of disease, report, the research of the interactional effect of AM fungi and PGPR and mechanism etc. are more, but can yet there are no open report with the Antagonistic Fungi of the collaborative disease-resistant short life of AM fungi for single AMF.
Summary of the invention
For in the prior art for the present situation of the research that report and prevention and control soil-borne disease Antagonistic Fungi are not arranged of the Antagonistic Fungi of the collaborative disease-resistant short life of AMF, the present invention aim to provide a kind of and the collaborative disease-resistant short life of AMF the Antagonistic Fungi bacillus amyloliquefaciens (
Bacillus amyloliquefaciens) and as the application in the control soil-borne diseases of vegetable Antagonistic Fungi, by verification experimental verification, adopt bacillus amyloliquefaciens provided by the invention (
Bacillus amyloliquefaciens) bacterium with antagonistic action has outstanding technique effect for the solution of tomato wilt, cucumber fusarium axysporum, Phytophthora capsici disease, have extensively and applicable value as control soil-borne diseases of vegetable Antagonistic Fungi.
The present invention adopts main technical scheme:
By taking pedotheque to carry out the antagonism screening in the farmland of planting for many years in the Fukang, Xinjiang suburb, obtain to have stronger bacteriostatic activity bacterial strain, through further screening, domestication breeding, obtain a strain and be numbered S-13.By obtaining bacterial strain being carried out morphological specificity and 16S rRNA sequencing and Phylogenetic Analysis, tentatively determined its classification position; Simultaneously the characteristics such as the bacteriostasis rate of the fermented liquid of this bacterium S-13, stability are conducted in-depth research, provide new selection as control soil-borne diseases of vegetable Antagonistic Fungi, its antagonistic action has outstanding technique effect for the solution of tomato wilt, cucumber fusarium axysporum, Phytophthora capsici disease and pepper anthracnose.
The Antagonistic Fungi genus bacillus of the collaborative disease-resistant short life of a kind of and AMF that the present invention specifically provides (
Bacillus amyloliquefaciens), by separating in the agricultural land soil of planting for many years in the Fukang, Xinjiang suburb, screen and cultivating, obtain a collection of microorganism strains as the control soil-borne diseases of vegetable, therefrom filter out the bacterial strain that a strain is numbered S-13, the control soil-borne diseases of vegetable has stable and significant effect, through the microbiology classification and identification, belong to bacillus amyloliquefaciens (
Bacillus amyloliquefaciens).
Concrete, the Antagonistic Fungi bacillus amyloliquefaciens of a kind of and the collaborative disease-resistant short life of AMF provided by the invention (
Bacillus amyloliquefaciens), called after S-13.This bacterial strain was preserved in the international depositary institution of budapest treaty microorganism before the applying date: China Committee for Culture Collection of Microorganisms common micro-organisms center (CGMCC).Address: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, postcode: 100101.Preservation date is on September 3rd, 2012, and preserving number is CGMCC No. 6491.Through microbiology be accredited as genus bacillus (
Bacillus amyloliquefaciens).This bacterial strain optimum growing condition is: 32 ° of C of temperature, and substratum is solid LB, culture condition: pH 7.0-7.2, time 14-16h; Bacterium colony median size, flat, has fold at surface drying, and the edge is irregular, and is opaque; Its thalline of microscopic examination is rod-short, gives birth in the gemma, does not expand; According to the 9th edition " uncle Jie Shi systematic bacteriology identification handbook " ("
Bergey, s Manual of Systematic Bacterio-logy") and " commonly used bacterial system identification handbook " the S-13 bacterial strain carried out morphology measure, Physiology and biochemistry detect determine the S-13 bacterial strain be genus bacillus (
Bacillus amyloliquefaciens) in the member.By BLAST homology comparison, the 16S rRNA sequence of bacterial strain S-13 is carried out in ncbi database after BLAST analyzes, the constructing system evolutionary tree, shown in accompanying drawing 2, bacterial strain S-13 with
Bacillus amyloliquefaciensGU323369.1 is in minimum branch, is its allied species; Bacterial strain 16S rRNA sequence is committed to
GeneBankMiddle acquisition gene accession number is: JN676212.Thereby with bacterial strain S-13 be defined as bacillus amyloliquefaciens (
Bacillus amyloliquefaciens).
Further, the invention provides the Antagonistic Fungi genus bacillus S-13(that works in coordination with disease-resistant short life with AMF
Bacillus amyloliquefaciens) as the application that prevents and treats soil-borne diseases of vegetable.
Simultaneously, the present invention proves, that provide and the Antagonistic Fungi genus bacillus S-13(collaborative disease-resistant short life of AMF
Bacillus amyloliquefaciens) as the application of tomato wilt, cucumber fusarium axysporum, the sick Antagonistic Fungi of Phytophthora capsici, have good antagonistic ability, wide spectrum antagonistic action.
The present invention further provides the preservation condition of bacterial classification, adopted medium component: peptone 1%(W/V), yeast extract paste 0.5%(W/V), sodium-chlor 1% (W/V), agar 2%(W/V), pH 7.0-7.2,121 ° of C sterilized 20 minutes.Culture condition: under 32 ° of C conditions on its LB medium slant, cultivated 14-16 hour, the aseptic skimmed milk of rear employing is protective material, cryopreservation after the vacuum lyophilization; Every day use is kept on the inclined-plane of LB substratum, and 4 ° of C Refrigerator stores are for subsequent use.
By implementing the concrete summary of the invention of the present invention, can reach following beneficial effect.
(1) the invention provides the genus bacillus CGMCC No. 6491 that separation obtains, it is a kind of bacillus amyloliquefaciens, application as the control soil-borne diseases of vegetable has higher value, especially can be for the Antagonistic Fungi of local habitat this Climate of isolated adaptation and growing environment for improving crop yield, reduce the agricultural chemicals usage quantity, make up good ecotope and have active effect.
(2) the present invention is directed to vegetable disease several frequently seen in the industrialized agriculture and filter out the genus bacillus CGMCC No. 6491 that a strain has better antagonistic ability, wide spectrum antagonistic action, foundation through 16S rRNA sequence amplification, comparison, evolutionary tree is initially identified as bacillus
Bacillus amyloliquefaciensThe allied species of GU323369.1.Antagonistic Fungi S-13 has broad-spectrum antifungal activity, can suppress the fungies such as the former bacterium of tomato wilt, Phytophthora capsici disease pathogen bacterium, pepper anthracnose pathogenic bacteria, cucumber fusarium axysporum pathogenic bacteria, performance is good under flat board face-off culture condition, growth for the pathogenic bacteria mycelia has obvious restraining effect, and Antagonistic Fungi S-13 is through microbiotic deciding to grow and studies show that after inducing, S-13 can have at different sites such as the root soil of plant, root, stem, leaves and surely grows, and is the good antagonistic strain of a strain.In order to have optimized its fermention medium and zymotechnique in from now on convenient, efficiently utilization, gemma rate and output are largely increased simultaneously, have active effect for industrialized development.This bacterial classification genus bacillus S-13 and AMF are as the Antagonistic Fungi of collaborative disease-resistant short life, has good effect, application as the control soil-borne diseases of vegetable has potential using value, particularly as the application of tomato wilt, cucumber fusarium axysporum, Phytophthora capsici disease, have good antagonistic ability, wide spectrum antagonistic action.
Description of drawings
Fig. 1 be shown as bacillus amyloliquefaciens (
Bacillus amyloliquefaciens) CGMCC No. 6491
Colonial morphology figure.
Fig. 2 be shown as bacillus amyloliquefaciens (
Bacillus amyloliquefaciens) CGMCC No. 6491
Growth curve chart.
Fig. 3 be shown as bacillus amyloliquefaciens (
Bacillus amyloliquefaciens) CGMCC No. 6491
16S rRNA phylogeny tree graph.
Fig. 4 is shown as a result figure of cucumber fusarium axysporum morbidity statistics.
Fig. 5 is shown as dull and stereotyped face-off and cultivates and the tomato seedling lab diagram.
Fig. 6 is shown as a result figure of tomato morbidity statistics.
Fig. 7 is shown as a result figure of the sick plant mortality statistics of Phytophthora capsici.
Embodiment
Below, for embodiment the present invention is described, still, the present invention is not limited to following embodiment.
The main raw and auxiliary material, reagent and the plant and instrument that relate among the present invention:
The phytopathogen that the present invention adopts: the tomato wilt bacterial strain (
Fusarium oxysporum), the former bacterium of cucumber fusarium axysporum (
Fusarium oxysporium f. sp. cucumerinum), the Phytophthora capsici pathogenic bacteria (
Phytophthora cap-sici Leonian) all provided by ACCC (Chinese agriculture microbial strains preservation center), the AM fungi that the present invention selects, those skilled in the art can obtain by public's channel.
Substratum and other material: substratum is selected: LB solid, liquid nutrient medium; Crop seed adopts tomato, cucumber, capsicum as representative test kind in the greengrocery.
Key instrument and reagent: MSSPX-250 type biochemical cultivation case, MLS-3020 high-pressure steam sterilizing pan, the single two-sided clean work station of SW-CJ-1F Type B, E360K whizzer, constant-temperature table HWY-100.PCR instrument Eppendorf No:5345, electrophoresis apparatus Bio-Rad Mode 200/2.0, gel imaging instrument United-Bio, GK-330C plus, the PCR premixed liquid (
TaKaRa Biotechnology), all the other reagent are analytical pure.
All bacterial classifications and the raw and auxiliary material selected among the present invention, and the spawn culture condition of selecting and method all be well known selecting, the % that relates among the present invention is weight percentage, unless otherwise indicated except.
Embodiment one: bacillus amyloliquefaciens (
Bacillus amyloliquefaciens) CGMCC No.
6491
Separation, screening and evaluation
(1) Isolation and screening of Antagonistic Fungi
From farmland sample, isolate many strains bacterium by the gradient dilution method, adopt dull and stereotyped face-off method, according to having or not inhibition zone to judge whether it has antagonistic action, preliminary screening goes out the bacterial strain that the former bacterium of tomato wilt, the former bacterium of cucumber fusarium axysporum, Phytophthora capsici pathogenic bacteria is had certain antagonistic ability.Bacterial strain with antagonistic action is carried out secondary face-off experiment, filter out the stronger bacterial strain of antagonistic ability in order to subsequent experimental.
Multiple sieve obtains the bacterial isolates that a strain has the former bacterium of good antagonism tomato wilt, the former bacterium of cucumber fusarium axysporum, the effect of Phytophthora capsici pathogenic bacteria, called after S-13, cultivate 72 hours its can significantly suppress the growth of pathogenic bacteria mycelia, verify that through many experiments its fungistatic effect is comparatively stable.
Referring to accompanying drawing 1, the S-13 bacterium of acquisition is in culture medium culturing, and this bacterial strain optimum growing condition is: temperature 32
° C, substratum is the LB solid medium, culture condition: pH7.0 cultivates 14-16h under 32 ° of C conditions of temperature; Bacterium colony median size, flat, has fold at surface drying, and the edge is irregular, and is opaque; Its thalline of microscopic examination is rod-short, gives birth in the gemma, does not expand; According to the 9th edition " uncle Jie Shi systematic bacteriology identification handbook " ("
Bergey, s Manual of Systematic Bacterio-logy") and " commonly used bacterial system identification handbook " the S-13 bacterial strain carried out morphology measure, Physiology and biochemistry detect determine the S-13 bacterial strain be genus bacillus (
Bacillus amyloliquefaciens) in the member.By above-mentioned the S-13 bacterial strain is carried out morphology mensuration, and shape, size, the biochemical reactions of bacterial strain S-13 detected, S-13 Biological Characteristics of Strain of the present invention is as shown in table 1.
(2) PCR amplification Antagonistic Fungi 16S rRNA sequence and order-checking thereof
The a small amount of single bacterium colony of picking is put into the EP pipe that fills 25 μ L sterilized waters, and 100 ° of C boil 8-10 min, rear mixture of ice and water 5 min that put into rapidly.Centrifugal 10000 r/min, 5 min, 4 ° of C preserve, and the time spent is got supernatant.Pcr amplification 16S rRNA sequence.
Primer 1 (27F): 5 '-TCCTCCGCTTATTGATATGC-3 ';
Primer 2 (1492R): 5 '-CAAACTTGGTCATTAGAGGA-3 '.
The pcr amplification reaction system is 50 μ L, contains 24 μ L premix Taq, primer 11 μ L, primer 21 μ L, template 2 μ L, sterilized water 22 μ L.Amplification condition: 94 ° of C 4 min, 94 ° of C 55 s, 53 ° of C 30 s, 72 ° of C 90 s, 30 circulations; 72 ° of C 7 min.Amplified production (about 1500 bp) is through 1% agarose gel electrophoresis isolation identification, and the PCR product directly carries out two-way order-checking.
(3) 16S rRNA sequence alignment and Phylogenetic Analysis
16S rRNA sequence and the nucleotide sequence in the GenBank database that order-checking is obtained carry out the BLAST analysis, therefrom obtain close 16S rRNA sequence, with Clustal X software and MEGA 4.1Neighbor-joining method constructing system evolutionary tree, referring to accompanying drawing 3.Bacterial strain 16S rRNA sequence is submitted to and is utilized the BankIt software among the GeneBank to submit to online, obtains the gene accession number.
In conjunction with Morphologic Characteristics and the physio-biochemical characteristics of S-13, determine that it is bacillus amyloliquefaciens (
Bacillus amyloliquefaciens) CGMCC No. 6491.
The 16S rRNA sequence of S-13 is carried out after BLAST analyzes the constructing system evolutionary tree in ncbi database.Shown in accompanying drawing 3, bacterial strain S-13 with
Bacillus amyloliquefaciensEvolutionary distance is the shortest between the GU323369.1, is
Bacillus amyloliquefaciensAllied species.In conjunction with Morphologic Characteristics and the physio-biochemical characteristics of S-13, determine that it is bacillus amyloliquefaciens (
Bacillus amyloliquefaciens).
Table 1: the physio-biochemical characteristics of bacterial strain S-13
[0033] embodiment two: bacillus amyloliquefaciens (
Bacillus amyloliquefaciens) somatomedin of CGMCC No.6491
Referring to following mode, but should according to genus bacillus provided by the invention (
Bacillus amyloliquefaciens) CGMCC No. 6491 characteristics determine the somatomedin that it is concrete.
Bacterial strain S-13 inoculation culture.The results are shown in Table 2.
Drawn the most suitable somatomedin of bacterial strain S-13 by table 3.
| Temperature (℃) | 4 | 15 | 25 | 30 | 32 | 35 |
| Growing state | — | + | + | +++ | ++++ | +++ |
| Temperature (℃) | 38 | 45 | 50 | ? | ? | ? |
| Growing state | ++ | + | — | ? | ? | ? |
| pH | 1 | 2 | 3 | 4 | 5 | 6 |
| Growing state | — | — | — | — | + | ++ |
| pH | 7 | 8 | 9 | 10 | ? | ? |
| Growing state | +++ | + | — | — | ? | ? |
| NaCl concentration | 0.5％ | 1％ | 2％ | 3％ | 4％ | 5％ |
| Growing state | + | ++ | +++ | +++ | +++ | + |
| NaCl concentration | 6％ | 7％ | 8％ | 9％ | 10％ | ? |
| Growing state | — | — | — | — | — | ? |
| Penbritin μ g/ |
50 | 60 | 70 | 80 | 90 | 100 |
| Growing state | — | — | — | — | — | — |
Table 2: temperature, pH, salt, microbiotic are to bacterial strain S-13 affects on the growth
Embodiment three: bacillus amyloliquefaciens (
Bacillus amyloliquefaciens) Study on Fermentation of CGMCC No. 6491
Antagonistic Fungi S-13 is a bacillus, in order better to use and bring into play its effect in production practice, we can by the optimization of substratum and culture condition, can make it produce a large amount of gemma by chance at expectation, not only be conducive to save the aftertreatment cost, more help the maintenance of its activity.Optimization by carbon and nitrogen sources, pH value etc. realizes sporiferous purpose.
(1) substratum, pH's determines
Press pH value 6.0,6.5,7.0,7.5,8.0, at 220 r/min, 30 ℃ of condition bottom fermentations of temperature draw the most suitable growth pH value.
(2) growth curve and culture condition
Table 3 pH optimum result
| ? | 6.0 | 6.5 | 7.0* | 7.5 | 8.0 |
| S-13 | 0.288 | 1.357 | 1.463 | 1.421 | 0.088 |
Referring to accompanying drawing 2 Antagonistic Fungi S-13 growth curves, and see Table 3 pH optimum result, drawing S-13 from above result is 16h as the best incubation time of seed, and the most suitable growth pH is 7.0, and the large volume production gemma time is 24h.
(3) optimization of Carbon and nitrogen sources
Seed liquid nutrient medium: white sugar 2.0%, yeast extract paste 0.5%, corn steep liquor 7.5%, KH
2PO
40.1%, K
2HPO
40.1%, MgSO
47H
2O 0.05%, continues to optimize on the basis of seed culture medium;
A. carbon source optimizing:
Carbon source is respectively glucose, sucrose, and Semen Maydis powder, Zulkovsky starch, other compositions are KH
2PO
40.1%, K
2HPO
40.1%, MgSO
47H
2O 0.05%, pH7.0;
B. nitrogenous source optimization:
Nitrogenous source is respectively peptone, yeast extract paste, (NH
4)
2SO
4, other compositions are tested with carbon source optimizing.Test is 3 repetitions.
Detect by different Carbon and nitrogen sources OD values, draw the suitableeest Carbon and nitrogen sources of bacterial strain.
The OD value: get fermented liquid 0.5mL, add 2.5M HCL 1mL, add the 13.5mL deionized water, mixing is used spectrophotometer, measures under the 540nm wavelength.
(4) fermention medium, carbon and nitrogen sources optimum result
Table 4 test design is optimized optimum carbon source-OD value in the fermention medium
| ? | Glucose | Sucrose | Semen Maydis powder | Zulkovsky starch |
| S-13 | 1.596 | 1.643 | 1.685* | 1.644 |
Table 5 test design is optimized optimum nitrogen source-OD value in the fermention medium
| ? | Peptone | Yeast extract paste | （NH 4） 2 SO 4 |
| S-13 | 1.476 | 1.621* | 1.395 |
Determine that according to above result optimum carbon source is Semen Maydis powder, optimum nitrogen source is yeast extract paste.
(5) 10L ferment tank result
Take corn steep liquor and Semen Maydis powder in the substratum of main raw material, fermentation condition is: 32 ℃ of temperature, rotating speed 250rpm, ventilation 1:0.3, the time is at 24h.Its result is as follows:
Table 6 10L fermenting experiment result
| Bacterial strain | Fermentation time | Extension rate | OD 540nm |
| S-13 | |
30 | 0.575 |
The microscopy result shows after the fermentation ends, almost all produces gemma, shows through thermal treatment coated plate counting, produces the gemma rate and reaches more than 95%, and through frozen dried, the gemma survival rate is higher than 90%.
Optimize fermentation results and show that bacterial strain S-13 is 16h as the best incubation time of seed, the most suitable growth pH is 7.0, and the large volume production gemma time is 24h; Determine that optimum carbon source is Semen Maydis powder, optimum nitrogen source is yeast extract paste, and the microscopy result shows after the 10L fermentation ends, almost all produces gemma, shows through thermal treatment coated plate counting, produces the gemma rate and reaches more than 95%, and through frozen dried, the gemma survival rate is higher than 90%.
Embodiment four: bacillus amyloliquefaciens (
Bacillus amyloliquefaciens) CGMCC No.
6491
With AM fungi synergistic result
1 experiment arranges:
1.1 every kind of pathogenic bacteria of this test arranges three experiments and processes, one group for only adding the control group of pathogenic bacteria, other one group for the treatment of group for adding pathogenic bacteria and AM fungi, all the other one group is the treatment group that adds simultaneously Antagonistic Fungi S-13 of the present invention and AM fungi, per 30 young plants are that an experiment is processed.Added up at the 50th day that has inoculated, calculate the crop mortality ratio.
1.2 pathogenic bacteria is processed and Antagonistic Fungi access of the present invention
Cultivate the crop seedling with the land for growing field crops soil after the sterilization at seedling bed, be divided into an AM fungi and do not add two groups of AM fungies, treat that seedling divides seedling when growing to a leaf one core, the root inoculation method is dipped in employing, the spore suspension that root is damaged a little the pathogenic bacteria of putting into 106 spore/mL soaks root inoculation 15min, then to plant diameter be 8 cm, be equipped with in the nutrition pot of sterilization land for growing field crops soil, every alms bowl one strain, and experimental group continues to add the AM fungi in nutrition pot.Cultivate in the greenhouse after 1 day, adopt filling root mode to add the Antagonistic Fungi suspension, the root irrigation group is pushed the soil around the root system aside, and pouring Antagonistic Fungi concentration is the bacteria suspension 10mL of 108cfu/mL, and all the other groups are then irrigated the clear water of same amount.
2 synergy research and results
2.2 to the research of Disease Resistance of Cucumber synergy
Referring to accompanying drawing 4 cucumber fusarium axysporum morbidity statistics figure as a result.In the collaborative research of cucumber seedling-raising, show inoculation cucumber fusarium axysporum pathogenic bacteria and 50 days " Invest, Then Investigate "s of Antagonistic Fungi: the mortality ratio of two control groups is all above 80%, minimum is the experimental group of inoculation Antagonistic Fungi S-13, cucumber plant mortality ratio about 30%, thus interpretation of result, Antagonistic Fungi S-13 of the present invention and AM fungi have preferably synergistic function aspect opposing, the control cucumber fusarium axysporum;
2.3 to the research of tomato disease resistance synergy
Cultivate and tomato seedling experiment and accompanying drawing 6 tomato morbidity statistics results referring to accompanying drawing 5 dull and stereotyped face-offs.In the collaborative research of tomato seedling, show at inoculating tomato Pathogen of Fusarium Wilt and 50 days " Invest, Then Investigate "s of Antagonistic Fungi:
The control group sickness rate surpasses 70%, and all the other group sickness rate are all lower, and the minimum experiment for adding Antagonistic Fungi S-13 is processed, and its sickness rate only is 2.4%.The antagonism synergy of tomato wilt is better with bacterial strain S-13 effect of the present invention, the effect that has significant collaborative anti-tomato wilt after the acting in conjunction of AM fungi.
2.4 the sick collaborative antagonism result of Phytophthora capsici
Referring to the sick plant mortality statistics of accompanying drawing 7 Phytophthora capsicis result.Close synergy research from the antagonism to the Phytophthora capsici disease, do not add the mortality ratio of processing pepper plant when investigation of Antagonistic Fungi all above 60%, the control group that does not add AM fungi and Antagonistic Fungi is up to more than 80%, and the experimental group sickness rate that adds Antagonistic Fungi is lower, sickness rate is lower than 20%, illustrates that Antagonistic Fungi and AM fungi can work in coordination with anti-Phytophthora capsici disease pathogen bacterium.
With the research of jointly growing seedlings of AM fungi, Antagonistic Fungi S-13 of the present invention and AM fungi have preferably synergy in opposing, control cucumber fusarium axysporum, tomato wilt, Phytophthora capsici aspect sick, can effectively reduce plant mortality ratio and sickness rate.Show genus bacillus provided by the invention (
Bacillus amyloliquefaciens) CGMCC No. 6491 as the control soil-borne diseases of vegetable application have potential using value, Antagonistic Fungi S-13 bacillus amyloliquefaciens (
Bacillus amyloliquefaciens) CGMCC No.6491 is as having good effect in the application of control in the soil-borne diseases of vegetable.
Claims (5)
1. the Antagonistic Fungi with the collaborative disease-resistant short life of AMF is characterized in that, Antagonistic Fungi be bacillus amyloliquefaciens (
Bacillus amyloliquefaciens) CGMCC No. 6491.
Antagonistic Fungi bacillus amyloliquefaciens as claimed in claim 1 (
Bacillus amyloliquefaciens) CGMCC No. 6491 is as the application in the control soil-borne diseases of vegetable.
Antagonistic Fungi bacillus amyloliquefaciens as claimed in claim 1 (
Bacillus amyloliquefaciens) CGMCC No. 6491 is as the application of tomato wilt Antagonistic Fungi.
Antagonistic Fungi bacillus amyloliquefaciens as claimed in claim 1 (
Bacillus amyloliquefaciens) CGMCC No. 6491 is as the application of cucumber fusarium axysporum Antagonistic Fungi.
Antagonistic Fungi bacillus amyloliquefaciens as claimed in claim 1 (
Bacillus amyloliquefaciens) CGMCC No. 6491 is as the application of the sick Antagonistic Fungi of Phytophthora capsici.
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